Amino Acids 1

Amino Acids
Karlheinz Drauz, Degussa AG, Hanau-Wolfgang, Germany
Ian Grayson, Degussa, Middlesbrough, United Kingdom
Axel Kleemann, Hanau, Germany
Hans-Peter Krimmer, Degussa AG, Hanau-Wolfgang, Germany
Wolfgang Leuchtenberger, Degussa AG, Düsseldorf, Germany
Christoph Weckbecker, Degussa AG, Hanau-Wolfgang, Germany

1. Introduction and History . . . . . . 1 3.2.17. l-Tryptophan . . . . . . . . . . . . . . 24

2. Properties . . . . . . . . . . . . . . . . 2 3.2.18. l-Tyrosine . . . . . . . . . . . . . . . . 24
2.1. Physical Properties and Structure 2 3.2.19. l-Valine . . . . . . . . . . . . . . . . . . 25
2.2. Chemical Properties . . . . . . . . . 4 4. Biochemical and Physiological
2.3. Important Amino Acids . . . . . . . 7 Significance . . . . . . . . . . . . . . . 25
2.3.1. Proteinogenic Amino Acids . . . . . 7 5. Uses . . . . . . . . . . . . . . . . . . . . 26
2.3.2. Other Important Amino Acids . . . . 9 5.1. Human Nutrition . . . . . . . . . . . 26
3. Industrial Production of Amino 5.1.1. Supplementation . . . . . . . . . . . . 28
Acids . . . . . . . . . . . . . . . . . . . 13 5.1.2. Flavorings, Taste Enhancers, and
3.1. General Methods . . . . . . . . . . . 14 Sweeteners . . . . . . . . . . . . . . . . 30
3.2. Production of Specific Amino Acids 15 5.1.3. Other Uses in Foodstuff Technology 31
3.2.1. l-Alanine . . . . . . . . . . . . . . . . . 15 5.2. Animal Nutrition . . . . . . . . . . . 32
3.2.2. l-Arginine . . . . . . . . . . . . . . . . 15 5.3. Pharmaceuticals . . . . . . . . . . . . 35
3.2.3. l-Aspartic Acid and Asparagine . . . 16 5.3.1. Nutritive Agents . . . . . . . . . . . . 36
3.2.4. l-Cystine and l-Cysteine . . . . . . . 17
5.3.2. Therapeutic Agents . . . . . . . . . . . 36
3.2.5. l-Glutamic Acid . . . . . . . . . . . . 17
5.4. Cosmetics . . . . . . . . . . . . . . . . 40
3.2.6. l-Glutamine . . . . . . . . . . . . . . . 18
5.5. Agrochemicals . . . . . . . . . . . . . 41
3.2.7. l-Histidine . . . . . . . . . . . . . . . . 19
5.5.1. Herbicides . . . . . . . . . . . . . . . . 41
3.2.8. l-Hydroxyproline . . . . . . . . . . . . 19
3.2.9. l-Isoleucine . . . . . . . . . . . . . . . 19 5.5.2. Fungicides . . . . . . . . . . . . . . . . 43
3.2.10. l-Leucine . . . . . . . . . . . . . . . . . 19 5.5.3. Insecticides . . . . . . . . . . . . . . . 45
3.2.11. l-Lysine . . . . . . . . . . . . . . . . . . 20 5.5.4. Plant Growth Regulators . . . . . . . 45
3.2.12. d,l-Methionine and l-Methionine . 21 5.6. Industrial Uses . . . . . . . . . . . . . 45
3.2.13. l-Phenylalanine . . . . . . . . . . . . . 21 6. Chemical Analysis . . . . . . . . . . 46
3.2.14. l-Proline . . . . . . . . . . . . . . . . . 23 7. Economic Significance . . . . . . . . 48
3.2.15. l-Serine . . . . . . . . . . . . . . . . . . 23 8. Toxicology . . . . . . . . . . . . . . . . 49
3.2.16. l-Threonine . . . . . . . . . . . . . . . 23 9. References . . . . . . . . . . . . . . . . 50

1. Introduction and History
The proteins, although they occur in an almost
infinite variety, are composed of a relatively
small number of basic building blocks, all α-
amino acids. In addition, the amino acids fulfill
certain regulatory functions in the metabolism
and are required for the biosynthesis of other
functional structures. This review is limited,
for the most part, to the protein-forming amino
acids, because they are by far the most widely
distributed in nature and are of considerable eco-
nomic interest.
The ca. 20 different α-amino acids found
in proteins are simple organic compounds, in
which an amino group and a side chain (R)
are attached alpha to the carboxyl function. The
R group may be aliphatic, aromatic, or hetero-
cyclic and may possess further functionality.

c 2006 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim

10.1002/14356007.a02 057.pub2
2 Amino Acids
Amino acid
Dipeptide
Protein (n > 50)
At present over 200 naturally occurring α-
amino acids are known [1–7]. Table 1 shows the
α-amino acids found in proteins, where they oc-
cur exclusively as the l-enantiomers. d-Amino
acids have been found only in the cell walls of
some bacteria, in peptide antibiotics, and in the
cell pools of some plants [10, 14, 15]. The l-ami-
no acids are commonly abbreviated with three-
letter or one-letter codes, as shown in Table 1 [4].
In addition, the abbreviation Glx or Z is used to
refer to either glutamic acid or glutamine, and
the abbreviation Asx or B is used to refer to ei-
ther aspartic acid or asparagine. More recently
History. The history of amino acid chemistry
began in 1806, when two French investigators,
Vauquelin and Robiquet, isolated asparagine
from asparagus juice. It was not until 1925 that
Schryver and Burton isolated threonine from
oat protein, the last of the ca. 20 protein-forming
amino acids to be discovered. Strecker synthe-
sized alanine in 1850 from acetaldehyde, ammo-
nia, and hydrogen cyanide. Escher established
the hypothesis of essential amino acids. Emil Fis-
cher discovered that the amino acids were build-
ing blocks of the proteins. Abderhalden synthe-
sized threonine from acrylic acid derivatives and
methanol. Rose et al. recognized threonine as
the last of the eight essential amino acids. d,l-
Methionine was produced industrially in Ger-
many in 1948, and in 1956 l-glutamic acid was
produced by fermentation in Japan.
Origin of Amino Acids. The first amino
acids were probably produced on the earth more
than 3×109 years ago via “prebiotic synthesis”
in the primordial atmosphere. The concept of
prebiotic synthesis is based on laboratory exper-
iments in which glycine, alanine, aspartic acid,
glutamic acid, and other compounds were pro-
duced by the action of an electrical discharge on
a simulated primordial atmosphere consisting of
methane, hydrogen, water, and ammonia [16].
Since then, traces of amino acids have been de-
tected in moon rocks, meteorites, and interstellar
space.
2. Properties
2.1. Physical Properties and Structure
α-Amino acids are nonvolatile, white, crys-
talline compounds with no defined melting
points. They are relatively stable on heating,
generally decomposing at 250–300 ◦ C. Both the
low volatility and the thermal stability result
from the low-energy dipolar structure (zwitter-
ion, inner salt, betaine), which the amino acids
assume in the solid state.
Evidence for this structure is provided by in-
frared and Raman spectra in which the bands
typical of -NH2 and -COOH moieties are ab-
sent. Equilibrium in solution also lies almost ex-
clusively on the side of the dipolar form; there-
fore, amino acids are insoluble in nonpolar sol-
vents and usually not very soluble in polar ones.
The only amino acids that exhibit any appre-
ciable solubility in alcohol are proline and hy-
droxyproline. Solubility in water depends on the
pH: the minimum is at the isoelectric point.
This solubility minimum at the isoelectric
point is quite useful for purifying and recrystal-
lizing amino acids. The analytical technique for
separating amino acid mixtures by electrophore-
sis is based on the fact that a specific amino acid
does not migrate in an electric field at its isoelec-
tric point, pI, a physical constant for each amino
acid.
The structures and physical properties of the
most important α-amino acids are given in Sec-
tion 2.3.1.
Stereochemistry. With the exception of
glycine, the simplest amino acid (R = H), all nat-
ural α-amino acids are chiral compounds occur-
ring in two enantiomeric (mirror-image) forms.
 Amino Acids 3
Table 1. Common amino acids of proteins
Trivial name IUPAC abbreviation One-Letter Protein Content, g/100 g
Code
[8] [10]
l-Alanine Ala A silk fibroin 25 29.7
l-Arginine Arg R salmin 87
edostin 17
wool 10
gelatin 8.3
rat liver histone 15.9
l-Asparagine Asn N
l-Aspartic acid Asp D edestin 12
hemoglobin 9–10
barley globulin 10.3
l-Cysteine Cys C wool keratin 11.9
human hair keratin 14.4
feather keratin 8.2
l-Cystine Cys-Cys, (Cys)2 , Cyss C
l-Glutamic acid Glu E gliadin 47
zein 31
wheat gliadin 39.2
maize zein 22.9
l-Glutamine Gln Q
Glycine Gly G gelatin 26
silk fibroin 44
l-Histidine His H hemoglobin 7
l-Hydroxyproline Hyp – gelatin 15
l-Isoleucine Ile I edestin 21
hemoglobin 29
serum proteins 20
oat globulin 4.3
beef serum albumin 2.6
l-Leucine Leu L edestin 21
hemoglobin 29
serum proteins 20
maize zeins 19
l-Lysine Lys K serum albumin 13
serum globulin 6
horse myoglobin 15.5
l-Methionine Met M egg albumin 5
casein 3 4.1
β-lactoglobulin 3.2
l-Phenylalanine Phe F zein 8
egg albumin 5 7.7
serum albumin 7.8
l-Proline Pro P gelatin 17 16.3
gliadin 13
salmin 6.9
casein 10.6
l-Serine Ser S silk fibroin 13 16.2
trypsinogen 16.7
pepsin 12.2
l-Threonine Thr T casein 4
human hair keratin 8.5
avidin 10.5
l-Tryptophan Trp W fibrin 3
egg lysozyme 10.6
l-Tyrosine Tyr Y fibrin 6
silk fibroin 13
papain 14.7
l-Valine Val V casein 8
beef sinew 17.4
beef aorta 17.6
4 Amino Acids
l-Amino acid d-Amino acid
The prefixes l and d express the absolute con-
figuration at the α-carbon atom by means of the
formal stereochemical relationship to l- or d-
glyceraldehyde, the reference substance intro-
duced by Emil Fischer in 1891. In addition to
the spacial representations shown above, the Fis-
cher projections are also universally recognized
and used:
l-Amino acid d-Amino acid
Polarimetric determination of the specific ro-
t Acidity and Basicity. The chemical proper-
tation [α]D can be used to differentiate between
the two enantiomers and to check their optical
t
purity. The molecular rotation [M ]D is less com-
mon:
Mr
[M ]tD = ·[α]tD
100
M r molecular mass; t temperature; D
589.3 nm (wavelength of the sodium D line).
Other methods for investigating the structure
of amino acid enantiomers include the Cotton
effect (change in molecular rotation as a func-
tion of the wavelength of plane-polarized light),
as reflected in optical rotational dispersion (re-
versal of the direction of the molecular rotation
at the wavelength of the absorption maximum),
and circular dichroism (differing absorption for
left- and right-handed circularly polarized light).
l-Amino acids exhibit a positive carbonyl Cot-
ton effect, d-amino acids a negative one.
Isoleucine, threonine, and hydroxyproline
each contain two chiral carbon atoms; therefore,
they appear in four stereoisomeric forms. Cys-
tine, which likewise contains two chiral carbons,
has only three stereoisomers: l-, d-, and meso-
cystine, the meso form having a plane of symme-
try. According to the Cahn–Ingold–Prelog rule
(R, S rule) [17], all proteinogenic l-α-amino
acids, with the exception of l-cysteine and l-
cystine, are S; the d-α-amino acids are R. Ac-
cording to this system, l-threonine, for exam-
ple, is termed (2S, 3R)-threonine. However, the
l- and d-descriptors are still in widespread use
for the simple l-α-amino acids.
Absorption Spectra. Aliphatic amino acids
exhibit no absorption in the UV region above
220 nm, with the exception of cystine (240 nm).
The aromatic amino acids, phenylalanine, tyro-
sine, and tryptophan, absorb between 250 and
300 nm [1, vol. 2]. The exact position of the max-
imum and the molar extinction coefficient  are
affected by the pH of the aqueous solution.
Two infrared bands are especially character-
−1 −
istic of amino acids: 1560–1600

 cm (–COO )
−1 +
and ca. 3070 cm −NH3 . These bands are
also evidence of the bipolar nature of the amino
acids.
2.2. Chemical Properties
ties of the α-amino acids are primarily the prop-
erties of the amino and carboxyl groups. Amino
acids react with strong acids as proton acceptors
(bases) and with strong bases as proton donors
(acids). In acidic medium they are present pre-
dominantly as cations; in basic medium they are
present predominantly as anions. Figure 1 shows
the titration curves of glycine, glutamic acid, and
lysine. The pK 1 and pK 2 values correspond to
the inflection points of the titration curve, the pH
value where the concentrations of the zwitterion
form and the cationic or anionic form are equal.
The pK 1 , pK 2 , and pI values can be calculated
from the titration curves with the Henderson–
Hasselbalch equation:
Amino acids with additional basic or acidic
groups (arginine, lysine, glutamic acid, cysteine)

exhibit additional pK values. Amino acids act as
buffers in the region of their pK values.
Figure 1. Titration curves of glycine, glutamic acid, and
lysine [9]
The pK 1 values show the amino acids to
be considerably stronger acids than acetic acid.
However, because of intramolecular protonation
of the amine moiety by the carboxyl group, aque-
ous solutions of amino acids are only weakly
acidic. The pH values of aqueous monoami-
nomonocarboxylic acids lie between 5.5 and 6.0.
Solutions of the acidic amino acids aspartic acid
and glutamic acid have pH values of ca. 2. The
weakly basic amino acid histidine has a pH of
7.5 in aqueous solution; the more strongly basic
amino acids lysine and arginine have pH values
of ca. 11–12. These differences in acidities and
basicities are utilized in the separation of amino
acid mixtures by ion-exchange chromatography
and electrophoresis.
Reactions. Because of their bifunctional and
sometimes trifunctional character, the α-amino
acids are capable of taking part in a variety of
chemical reactions. Comprehensive treatments
of these may be found in monographs and re-
views [1, 9–12, 18].
The introduction of groups protecting the
amino, carboxyl, and side-chain functions is es-
pecially important for peptide synthesis [13].
Additionally, α-amino acids play a prominent
role as intermediates in the synthesis of hetero-
Amino Acids 5
cycles [19]. The chiral α-amino acids and their
derivatives are inexpensive and, for the most
part, readily available synthons for numerous
natural products and pharmaceuticals. In many
cases optically active amino acids can also be
used to induce chirality during the course of a
synthetic process [20].
α-Amino acids form chelate-like complexes
with heavy metal ions. The best known are the
dark blue, easily crystallized copper chelates:
Bis(glycinato) copper(II) hydrate
This complex formation can be used to pro-
tect both the α-amino and the carboxyl group
during synthesis of -N-acetyl derivatives or
carbamates of lysine or other side-chain deriva-
tives. In these copper or cobalt complexes the
α-carbon atom is activated sufficiently to react
with aldehydes. A well-known example is the al-
kaline condensation of the glycine–copper com-
plex with acetaldehyde, resulting in threonine.
Especially important is the reaction of α-amino
acids with ninhydrin to form a blue-violet dye,
the basis of a sensitive optical method for de-
tecting amino acids (see Chap. 6).
Free amino acids react with nitrous acid to
yield α-hydroxycarboxylic acids, with retention
of configuration. Volumetric measurement of the
nitrogen gas set free is the basis of Van Slyke’s
method for the quantitative analysis of amino
acids. Reaction of amino acid esters with ni-
trous acid gives the acid-labile diazocarboxylic
acid esters. Treatment of N-alkyl- or N-arylami-
no acids with nitrous acid yields the N-nitroso
derivatives, which can be dehydrated to syd-
nones in the presence of acetic anhydride:
6 Amino Acids

α-Amino acids have found use as organocat- with anhydrous alcohol in the presence of anhy-
alysts for asymmetric organic reactions, for ex- drous hydrogen chloride. The initial product is
ample asymmetric versions of the aldol or the the hydrochloride of the ester, which is liberated
Mannich reactions [21]. The amino acids most by addition of base. On standing or warming,
often employed are l- or d-proline, their deriva- the free esters of α-amino acids can eliminate
tives, or short-chain peptides based on proline. alcohol to form 2,5-diketopiperazines:
Oxidizing agents attack the amino group,
converting the amino acid into an iminocarbox-
ylic acid. These are unstable and either hydro-
lyze to α-oxocarboxylic acids or decompose, af-
ter decarboxylation, into ammonia and the alde-
hyde containing one carbon atom less. Dike- Amino acid esters are useful intermediates for
tones, triketones, N-bromosuccinimide, or silver peptide synthesis because the carboxyl function
oxide may serve as the oxidizing agent. The best is protected. The esters may be converted to ami-
known example is the reaction with ninhydrin. no alcohols by treatment with a strong reducing
Oxidative deamination also can be carried out agent, such as lithium aluminum hydride.
enzymatically with d- or l-amino acid oxidases. Several cyclic derivatives of the α-amino
This also proceeds via the α-iminocarboxylic acids are of industrial importance. The hydan-
acids, which subsequently are hydrolyzed to α- toins or imidazolidine-2,4-diones, which have
oxo acids: been used as intermediates in the synthesis of
α-amino acids, are most conveniently prepared
by treatment of aldehyde cyanohydrins with am-
monium carbonate or urea. They also may be
obtained by reacting amino acids with cyanates
or isocyanates, the reaction proceeding via the
ureidocarboxylic acid (hydantoic acid deriva-
tive):
This enantioselective enzymatic oxidative
deamination is the basis of analytical methods
for the determination of amino acid enantiomers.
The N-acylation of α-amino acids with
acyl chlorides or anhydrides under Schotten-
Baumann conditions produces N-acyl-α-amino
acids, which have numerous uses. For exam-
ple, the N-acetyl derivatives of d,l-amino acids
(e.g., alanine, valine, methionine, phenylala-
nine, tryptophan) are intermediates in the pro-
duction of l-amino acids by enzymatic resolu-
tion using aminoacylases (see Section 3.1). The thiohydantoins are obtained by the reaction
Amino acids acylated with naturally occur- with isothiocyanates.
ring fatty acid residues are used industrially as Dehydration of N-acylamino acids with
biodegradable surfactants. acetic anhydride or carbodiimide yields 1,3-
If free amino acids are heated above 200 ◦ C, oxazolin-5-ones (azlactones):
especially in the presence of soda lime or metal
ions, they readily decarboxylate to form amines.
The enzymatic decarboxylation of amino acids
gives biogenic amines. Some of these amines are
physiologically active neurotransmitters (his-
tamine, tyramine, dopamine, serotonin).
The esters are usually prepared by direct es- Racemization occurs readily during the reaction.
terification, e.g., reaction of the α-amino acid The azlactones are intermediates in the synthesis
of amino acids.

The N-carboxylic acid anhydrides (Leuchs
anhydrides, 1,3-oxazolidine-2,5-diones) are of-
ten used in peptide synthesis, especially to pre-
pare poly-α-amino acids. These reactive amino
acid derivatives are obtained by treating the ami-
no acid with phosgene in the presence of tertiary
amines
or by elimination of benzyl chloride from N-
benzyloxycarbonylamino acid chlorides.
2.3. Important Amino Acids
2.3.1. Proteinogenic Amino Acids
L-Alanine [56-41-7], 2-aminopropionic
acid, C3 H7 NO2 , M r 89.09, mp 314 ◦ C (de-
25
comp.), [α]D + 14.47◦ (c = 10.03 in 6 M HCl),
solubility 16.51 (25 ◦ C) g/100 g H2 O, pI 6.01,
dissociation constants: pK 1 2.34, pK 2 9.69.
L-Arginine [74-79-3], 2-amino-5-guani-
dinovaleric acid, 2-amino-5[(aminoimino-
methyl)amino]pentanoic acid, C6 H14 N4 O2 , M r
174.20, mp 244 ◦ C (decomp.), [α]25
D + 27.58
◦ 25
◦
(c = 2 in 6 M HCl), solubility 14.87 (20 C)
g/100 g H2 O, pI 10.76, dissociation con-
stants: pK 1 2.01, pK 2 9.04 (α-NH2 ), pK 3
12.48 (guanidyl). Hydrochloride: [1119-34-2],
C6 H15 ClN4 O2 , M r 210.66, mp 220 ◦ C (de-
25
comp.), [α]D + 11.7◦ (c = 5 in H2 O), solubility
◦
75.1 (20 C) g/100 g H2 O.
L-Asparagine [70-47-3], 2-aminosuccin-
amic acid, 2,4-diamino-4-oxobutanoic acid,
C4 H8 N2 O3 , M r 132.13, mp 236 ◦ C (decomp.),
20
[α]D + 32.6◦ (c = 1 in 0.1 M HCl), solubility
3.11 (28 ◦ C) g/100 g H2 O, pI 5.41, dissociation
constants: pK 1 2.02, pK 2 8.8.
Amino Acids 7
L-Aspartic acid [56-84-8], aminosuccinic
acid, 2-amino-1,3-butanedioic acid, C4 H7 NO4 ,
M r 133.10, mp 270 ◦ C (decomp., sealed tube),
25
[α]D + 25.4◦ (c = 2 in 5 M HCl), solubility 0.5
(25 ◦ C) g/100 g H2 O, pI 2.98, dissociation con-
stants: pK 1 2.1, pK 2 3.86 (β-COOH), pK 3 9.82.
L-Cysteine [52-90-4], 2-amino-3-mer-
captopropionic acid, 3-mercaptoalanine,
C3 H7 NO2 S, M r 121.16, mp 240 ◦ C (decomp.),
25
[α]D + 9.7◦ (c = 8 in 1 M HCl), solubility 28
(25 ◦ C) g/100 mL solution, 16 (20 ◦ C) g/100 g
H2 O, pI 5.02, dissociation constants: pK 1 1.71,
pK 2 8.27 (–SH), pK 3 10.78. Hydrochloride
monohydrate: [7048-04-6], C3 H10 ClNO3 S,
M r 175.64, mp (anhydr.) 178 ◦ C (decomp.),
25
[α]D + 6.53◦ (calculated as cysteine) (c = 2 in
5 M HCl), solubility >100 (20 ◦ C) g/100 g H2 O.
L-Cystine [56-89-3], 2,2 -diamino-3, 3 -
dithiobis(propionic acid), 3,3 -dithiobis(2-ami-
nopropanoic acid), C6 H12 N2 O4 S2 , M r 240.30,
mp 260◦ C (decomp.), [α]D −212◦ (c = 1 in 1 M
HCl), solubility 0.011 (25 ◦ C) g/100 g H2 O, pI
5.02, dissociation constants: pK 1 1.04, pK 2 2.05
(-COOH), pK 3 8.0 (–NH2 ), pK 4 10.25 (–NH2 ).
L-Glutamic acid [56-86-0], 2-amino-
glutaric acid, 2-amino-1,4-pentanedioic acid,
C5 H9 NO4 , M r 147.13, mp 224–225 ◦ C (de-
25
comp.), [α]D + 31.5◦ (c = 2 in 5 M HCl, 20 ◦ C),
solubility = 0.843 (25 ◦ C) g/100 g H2 O, pI 3.08,
dissociation constants: pK 1 2.1, pK 2 4.07, pK 3
9.47.
8 Amino Acids

L-Glutamine [56-85-9], 2-amino-

glutaramic acid, 2,5-diamino-5-oxopentanoic
acid, C5 H10 N2 O3 , M r 146.15, mp 185-6 ◦ C
25
(decomp.), [α]D + 31.8◦ (c = 2 in 1 M HCl),
solubility 3.6 (19 ◦ C) g/100 g H2 O, pI 5.65, L-Leucine [61-90-5], 2-amino-4-methyl-
dissociation constants: pK 1 2.17, pK 2 9.13. valeric acid, 2-amino-4-methylpentanoic
acid, 2-aminoisocaproic acid, C6 H13 NO2 ,
M r 131.18, mp 293-5, 314-5 ◦ C (decomp.),
25
[α]D −10.9◦ (c = 0.52 in H2 O), solubility 2.19
(25 ◦ C) g/100 g H2 O, pI 5.98, dissociation con-
stants: pK 1 2.36, pK 2 9.6.
Glycine [56-40-6], aminoacetic acid,
C2 H5 NO2 , M r 75.07, mp 262, 292 ◦ C (de-
comp.), solubility 24.99 (25 ◦ C) g/100 g H2 O,
pI 6.06, dissociation constants: pK 1 2.35, pK 2
9.13. L-Lysine [56-87-1], 2,6-diaminohexanoic
acid, 2,6-diaminocaproic acid, C6 H14 N2 O2 , M r
25
146.19, mp 225-5 ◦ C (decomp.), [α]D + 25.9◦
(c = 2 in 5 M HCl), solubility >100 (25 ◦ C)
L-Histidine [71-00-1], α-amino-1H-imida-
g/100 g H2 O, pI 9.47, dissociation constants:
zole-4-propionic acid, 1H-imidazole-4-alanine,
pK 1 2.18, pK 2 8.95 (α-NH2 ), pK 3 10.53.
C6 H9 N3 O2 , M r 155.16, mp 277, 287 ◦ C (de-
25 Hydrochloride: [657-27-2], C6 H15 ClN2 O2 , M r
comp.), [α]D + 13.0◦ (c = 1 in 6 M HCl), solu- 182.65, mp 253-6 ◦ C (decomp.), solubility 72.5
bility 4.29 (25 ◦ C) g/100 g H2 O, pI 7.64, dis- (25 ◦ C) g/100 mL solution.
sociation constants: pK 1 1.77, pK 2 6.1 (im-
idazolyl), pK 3 9.18. Hydrochloride monohy-
drate: [5934-29-2], C6 H12 ClN3 O3 , M r 209.63,
mp 259 ◦ C (decomp.), solubility 16.99 (20 ◦ C)
g/100 g H2 O. L-Methionine [63-68-3], 2-amino-4-
(methylthio)butyric acid, 2-amino-4-(methyl-
thio)butanoic acid, C5 H11 NO2 S, M r 149.21,
28
mp 283 ◦ C (decomp.), [α]D + 23.4◦ (c = 5 in
3 M HCl), solubility 5.37 (20 ◦ C) g/100 g H2 O,
L-Hydroxyproline [51-35-4], trans-4-hy- pI 5.74, dissociation constants: pK 1 2.28, pK 2
droxy-2-pyrrolidinecarboxylic acid, C5 H9 NO3 , 9.21.
22
M r 131.13, mp 274 ◦ C (decomp.), [α]D −75.2◦
◦
(c = 2 in H2 O), solubility 36.11 (25 C) g/100 g
H2 O, pI 5.82, dissociation constants: pK 1 1.92,
pK 2 9.73.
L-Phenylalanine [63-91-2], 2-amino-
3-phenylpropionic acid, α-aminobenzenepro-
panoic acid, C9 H11 NO2 , M r 165.19, mp 283-
20
4 ◦ C (decomp.), [α]D −35.1◦ (c = 2 in H2 O),
◦
L-Isoleucine [73-32-5], 2-amino-3-methyl- solubility 2.965 (25 C) g/100 g H2 O, pI 5.48,
valeric acid, 2-amino-3-methylpentanoic acid, dissociation constants: pK 1 1.83, pK 2 9.13.
C6 H13 NO2 , M r 131.18, mp 285-6 ◦ C (de-
25
comp.), [α]D + 40.6◦ (c = 2 in 6 M HCl), solu-
bility 4.117 (25 ◦ C) g/100 g H2 O, pI 6.02, dis-
sociation constants: pK 1 2.36, pK 2 9.68.
L-Proline [147-85-3], 2-pyrrolidinecar-
boxylic acid, 2-carboxypyrrolidine, C5 H9 NO2 ,
 Amino Acids 9

M r 115.13, mp 220–222 ◦ C (decomp.),

25
[α]D −85.0◦ (c = 1 in H2 O), solubility 162.3
(25 ◦ C) g/100 g H2 O, pI 6.3, dissociation con-
stants: pK 1 2.0, pK 2 10.6.
L-Valine [72-18-4], 2-amino-3-methylbu-
tyric acid, 2-aminoisovaleric acid, C5 H11 NO2 ,
20
M r 117.15, mp 315 ◦ C (decomp.), [α]D +26.7◦
(c = 3.4 in 6 M HCl), solubility 8.85 (25 ◦ C)
L-Serine [56-45-1], 2-amino-3-hydroxy- g/100 g H2 O, pI 5.96, dissociation constants:
propionic acid, 2-amino-3-hydroxypropanoic pK 1 2.32, pK 2 9.62.
acid, C3 H7 NO3 , M r 105.09, mp 228 ◦ C (de-
26
comp.), [α]D −6.8◦ (c = 10 in H2 O), solubility
◦
35.97 (20 C) g/100 g H2 O, pI 5.68, dissociation
constants: pK 1 2.21, pK 2 9.15.

2.3.2. Other Important Amino Acids

L-Threonine [72-19-5], 2-amino-3-hy- β-Alanine [107-95-9], 3-aminopropionic

droxybutyric acid, 2-amino-3-hydroxybutanoic acid, C3 H7 NO2 , M r 89.09, dissociation con-
acid, C4 H9 NO3 , M r 119.12, mp 253 ◦ C (de- stant: pK 1 3.6, occurrence: apple, constituent of
26 pantothenic acid, carnosine, anserine.
comp.), [α]D −28.6◦ (c = 2 in H2 O), solubility
◦
9.03 (20 C) g/100 g H2 O, pI 6.16, dissociation
constants: pK 1 2.71, pK 2 9.62.

D-Alanine [338-69-2], 2-aminopropionic

L-Tryptophan [73-22-3], 2-amino-3-(3 -
indolyl)propionic acid, α-amino-1H-indole-3-
propanoic acid, C11 H12 N2 O2 , M r 204.23, mp
25
290–292 ◦ C, 281 ◦ C (decomp.), [α]D −32.15◦
◦ pI 6.11, dissociation constants: pK 1 2.35, pK 2
(c = 1 in H2 O), solubility 1.14 (25 C) g/100 g
H2 O, pI 5.88, dissociation constants: pK 1 2.38,
pK 2 9.39.
acid, d-Ala, C3 H7 NO2 , M r 89.09, application:
LHRH-antagonists, e.g., Abarelix (antineoplas-
tic) [22].
D,l-Alanine [302-72-7], 2-aminopropionic
acid, C3 H7 NO2 , M r 89.09, mp 295 ◦ C (de-
comp.), solubility 16.72 (25 ◦ C) g/100 g H2 O,
9.87.
α-Aminoisobutyric acid [62-57-7], 2-
amino-2-methylpropionic acid, Aib, C4 H9 NO2 ,
M r 103.12, application: growth hormone secre-
tagogues, e.g., MK-0677 [23].

L-Tyrosine [60-18-4], 2-amino-3-

(4-hydroxyphenyl)propionic acid, [3-(4-
hydroxyphenyl)]alanine, 2-amino-3-(p-hy-
droxyphenyl)propionic acid, α-amino-4-hy- L-α-Aminobutyric acid [1492-24-6], 2-
droxybenzenepropanoic acid, C9 H11 NO3 , M r aminobutyric acid, l-Abu, C4 H9 NO2 , M r
181.19, mp 342-4 (sealed tube), 297–298 ◦ C 103.12, application: Levetiracetam (anticonvul-
25
(decomp.), [α]D −7.27◦ (c = 4 in 6 M HCl), sant) [24].
solubility 0.045 (25 ◦ C) g/100 g H2 O, pI 5.63,
dissociation constants: pK 1 2.2, pK 2 9.11, pK 3
10.07 (–OH).
10 Amino Acids

γ-Aminobutyric acid (GABA) [56-12-2], Creatine [57-00-1], N-(aminoimino-

4-aminobutyric acid, C4 H9 NO2 , M r 103.12, oc- methyl)-N-methylglycine, N-methylguanetic
currence: citrus fruits, sugar beet, brain. acid, C4 H9 N3 O2 , M r 131.14, occurrence: mus-
cle of vertebrates.

D,L-Aspartic acid [617-45-8], ami-

nosuccinic acid, 2-amino-1,3-butanedioic acid,
C4 H7 NO4 , M r 133.10, mp 275 ◦ C (decomp.,
sealed tube), solubility 0.775 (25 ◦ C) g/100 g D-Cyclohexylalanine [58717-02-5], 2-
H2 O, pI 2.98, dissociation constants: pK 1 2.1, amino-3-cyclohexylpropionic acid, d-Cha,
pK 2 3.86 (β-COOH), pK 3 9.82. C9 H17 NO2 , M r 171.25, application: (thrombin
inhibitors) [27].
L-Azetidine-2-carboxylic acid [2133-34-
8], C4 H7 NO2 , M r 101.10, application: Xime-
lagatran (thrombin inhibitor) [25].

Betaine [107-43-7], carboxymethyl-tri-
methyl ammonium betaine, C5 H11 NO2 , M r
117.15, occurrence: sugar beet.
(3S,4aS,8aS)-Decahydroisoquinolinecar-
boxylic acid [115238-58-9], decahydroiso-
quinoline-3-carboxylic acid, C10 H17 NO2 , M r
183.26, application: Nelfinavir (antiviral) [28].

L-2,3-Diaminopropionic acid [4033-39-

L-Carnitine [541-15-1], (3-carboxy-2- 0], l-Dap, C3 H8 N2 O2 , M r 104.11, application:
hydroxypropyl)trimethyl ammonium betaine, Imidapril (antihypertensive) [29].
C7 H15 NO3 , M r 161.2, occurrence: Lys metabo-
lite, muscle.

l-3,4-Dihydroxyphenylalanine (DOPA)
[59-92-7], 2-amino-3-(3,4-dihydroxyphe-
nyl)propionic acid, C9 H11 NO4 , M r 197.17,
L-Citrulline [372-75-8], 2-amino-5-
occurrence: Tyr metabolite, faba bean.
ureidopentanoic acid, C6 H13 N3 O3 , M r 175.19,
25
mp 234–237 ◦ C , 222 ◦ C (decomp.), [α]D +
◦
24.2 (c = 2 in 5 M HCl), solubility 10.3 (20 ◦ C)
g/100 g H2 O, pI 5.92, dissociation constants:
pK 1 2.43, pK 2 9.41; occurrence: urea cycle,
watermelon. D-Glutamic acid [6893-26-1], 2-ami-
noglutaric acid, 2-amino-1,4-pentanedioic acid,
d-Glu, C5 H9 NO4 , M r 147.12, application:
Spiroglumide (CCK-B-antagonist) [30].

L-Homocysteine [6027-13-0], 2-amino-4-

D-Citrulline [13594-51-9], 2-amino-5- mercaptobutyric acid, C4 H9 NO2 S, M r 135.18,
ureidopentanoic acid, d-Cit, C6 H13 N3 O3 , M r occurrence: Met metabolite, mushrooms, appli-
175.19, application: Cetrorelix (antineoplastic) cation: Omapatrilat (ACE inhibitor) [31].
[26].

D-p-Hydroxyphenylglycine [22818-40-
2], amino-(4-hydroxyphenyl)acetic acid, d-
Phg(OH), C8 H9 NO3 , M r 167.15, application:
Amoxicillin (antibiotic) [32].
L-5-Hydroxytryptophan [4350-09-8], 2-
amino-3-(5-hydroxy-1H-indol-3-yl)propionic
acid, C11 H12 N2 O3 , M r 220.22, occurrence:
serotonin precursor.
D,L-Isoleucine [443-79-8], 2-amino-3-
methylvaleric acid, 2-amino-3-methylpentanoic
acid, C6 H13 NO2 , M r 131.18, mp 292 ◦ C (de-
comp.), solubility 2.011 (25 ◦ C) g/100 g H2 O,
pI 6.04, dissociation constants: pK 1 2.32, pK 2
9.76.
D,L-Leucine [328-39-2], 2-amino-4-
methylvaleric acid, 2-amino-4-methylpentanoic
acid, 2-aminoisocaproic acid, C6 H13 NO2 , M r
131.18, mp 293-5, 332 ◦ C (decomp.), solubility
1.00 (25 ◦ C) g/100 g H2 O, pI 6.04, dissociation
constants: pK 1 2.33, pK 2 9.74.
L-tert-Leucine [20859-02-3], 2-amino-3,3-
dimethylbutyric acid, l-Tle, C6 H13 NO2 , M r
131.18, application: div. pharmaceuticals [33].
D,L-Lysine hydrochloride [70-53-1],
C6 H15 ClN2 O2 , M r 182.65, mp 264 ◦ C (de-
comp.), solubility 35.98 (20 ◦ C) g/100 g H2 O.
D,L-Methionine [59-51-8], 2-amino-4-
(methylthio)butyric acid, 2-amino-4-(methyl-
thio)butanoic acid, C5 H11 NO2 S, M r 149.21,
Amino Acids 11
mp 281 ◦ C (decomp.), solubility 3.35 (25 ◦ C)
g/100 g H2 O, pI 5.74, dissociation constants:
pK 1 2.28, pK 2 9.21.
S-Methyl-L-cysteine [1187-84-4],2-ami-
no-3-methylthiopropionic acid l-Cys(Me),
C4 H9 NO2 S, Mr 135.18, application:
Kynostatin-272 (antiviral) [34].
L-S-Methylmethionine (vitamin U) [4727-
40-6], 3-amino-3-carboxy-propyl)-dimethyl
sulfonium, C6 H13 NO2 S, M r 163.24, occur-
rence: cabbage, asparagus.
D-3-(2
-Naphthyl)-alanine [76985-09-6],
2-amino-3-naphthalen-2-ylpropionic acid, d-
Nal, C13 H13 NO2 , M r 215.25, application:
LHRH-antagonists, e.g., Abarelix (antineoplas-
tic) [35].
D-Ornithine [70-26-8], 2,5-diaminovaleric
acid, C5 H12 N2 O2 , M r 132.16, mp 140 ◦ C (de-
25
comp.), [α]D + 16.5◦ (c = 4.6 in H2 O), sol-
ubility, pI 9.7, dissociation constants: pK 1
1.94, pK 2 8.65 (a-NH), pK 3 10.76; occur-
rence: urea cycle, shark liver, application: Atosi-
ban (tocolytic) [36]. Hydrochloride: [3184-13-
2], C5 H13 ClN2 O2 , M r 168.6, mp 215 ◦ C (de-
25
comp.), [α]D + 28.3◦ (calculated as ornithine)
(c = 2 in 5 M HCl), solubility 54.36 (20 ◦ C)
g/100 g H2 O.
D-Penicillamine [52-67-5], 2-amino-3-
mercapto-3-methylbutyric acid, C5 H11 NO2 S,
M r 149.21, occurrence: hydrolysis product of
penicillin.
12 Amino Acids
D-Phenylalanine [673-06-3], 2-amino-
3-phenylpropionic acid, α-aminobenzenepro-
panoic acid, d-Phe, C9 H11 NO2 , M r 165.19, ap-
plication: Nateglinide (antidiabetic) [38].
D,L-Phenylalanine [150-30-1], 2-ami-
no-3-phenylpropionic acid, α-aminobenzene-
propanoic acid, C9 H11 NO2 , M r 165.19, mp
284–288 ◦ C, 320 ◦ C (decomp.), solubility 1.29
(25 ◦ C) g/100 g H2 O, pI 5.91, dissociation con-
stants: pK 1 2.58, pK 2 9.24
D-p-Cl-Phenylalanine [14091-08-8], 2-
amino-3-(4-chlorophenyl)propionic acid, d-
Phe(Cl), C9 ClH10 NO2 , M r 199.63, application:
LHRH antagonists, e.g., Abarelix (antineoplas-
tic) [39].
L-p-NO2 -Phenylalanine [949-99-5], 2-
amino-3-(4-nitrophenyl)propionic acid, l-
Phe(NO2 ), C9 H10 N2 O4 , M r 210.17, applica-
tion: Zolmitriptan (antimigraine) [40].
S-Phenyl-L-cysteine [34317-61-8], 2-
amino-3-phenylthiopropionic acid, l-Cys(Ph),
C9 H11 NO2 S, M r 197.61, application: Nelfi-
navir (antiviral) [41].
D-Phenylglycine [875-74-1], 2-amino-
phenyl-acetic acid, d-Phg, C8 H9 NO2 , M r
151.16, application: Ampicillin (antibiotic)
[42].
L-Pipecolic acid [3105-95-1], piperidine-
2-carboxylic acid, l-Pec, C6 H11 NO2 , M r
129.16, occurrence: legumes, metabolite of Lys,
application: Ropivacaine (local anesthetic) [43].
L-Piperazinecarboxylic acid [147650-70-
2], piperazine-2-carboxylic acid, C5 H10 N2 O2 ,
M r 130.15, application: Indinavir (antiviral)
[44].
D-Piperidine-3-carboxylic acid [25137-00-
2], nipecotic acid, C6 H11 NO2 , M r 129.7, appli-
cation: Tiagabine (anticonvulsant) [45].
D,l-Proline [609-36-9], 2-pyrrolidinecar-
boxylic acid, 2-carboxypyrrolidine, C5 H9 NO2 ,
M r 115.13, mp 205 ◦ C (decomp.), pI 6.3, disso-
ciation constants: pK 1 2.0, pK 2 10.6.
D-Proline [344-25-2], 2-pyrrolidinecar-
boxylic acid, 2-carboxypyrrolidine, d-Pro,
C5 H9 NO2 , M r 115.13, application: Eletriptan
(antimigraine) [46].
D-3-(3
-Pyridyl)-alanine [70702-47-5],
2-amino-3-pyridin-3-ylpropionic acid, d-Pal,
C8 H10 N2 O2 , M r 166.18, application: LHRH
antagonist, e.g. Abarelix (antineoplastic) [47].
L-Pyrrolysine [448235-52-7], N 6 -[(3R)-
1,5-didehydro-3-methyl-d-prolyl]- l-lysine,
C12 H21 N3 O3 , M r 255.31, the 22nd natural ami-
no acid to be discovered, occurrence: archaea as
part of methane-producing enzymes [37].

L-Saccharopine [997-68-2], 2-(5 -ami-

no-5 -carboxypentylamino)pentanedioic acid,
C11 H20 N2 O6 , M r 276.27, occurrence: baker s
and brewer s yeast.
L-Selenocysteine [10236-58-5], 3-selenyl-
l-alanine, Sec, U, C3 H7 NO2 Se, M r 168.05, the
21st natural amino acid to be discovered [48].
Often found in the form of its dimer, R,R-
selenocystine [29621-88-3], C6 H12 N2 O4 Se2 ,
M r 334.09.
L-Selenomethionine [3211-76-5], (S)-
2-amino-4-(methylseleno)butanoic acid, (S)-
2-amino-4-(methylseleno)butanoic acid,
C5 H11 NO2 Se, M r 196.11, occurrence: plant
tissue, application: dietary supplement [49].
D,l-Serine [302-84-1], 2-amino-3-hydroxy-
propionic acid, 2-amino-3-hydroxypropanoic
acid, C3 H7 NO3 , M r 105.09, mp 246 ◦ C (de-
comp., sealed tube), solubility 5.023 (25 ◦ C)
g/100 g H2 O, pI 5.68, dissociation constants:
pK 1 2.21, pK 2 9.15.
D-Serine [312-84-5], 2-amino-3-hydroxy-
propionic acid, 2-amino-3-hydroxypropanoic
acid, d-Ser, C3 H7 NO3 , M r 105.09, application:
d-cycloserine (cognition enhancer) [50].
L-Thiazolidine-4-carboxylic acid [34292-
47-7], l-Tia, C4 H7 NO2 S, M r 133.16, applica-
tion: Kynostatin-272 (antiviral) [34].
D,L-Threonine [80-68-2], 2-amino-3-hy-
droxybutyric acid, 2-amino-3-hydroxybutanoic
acid, C4 H9 NO3 , M r 119.12, mp 234-5 ◦ C (de-
comp.), solubility 20.5 (25 ◦ C) g/100 g H2 O,
Amino Acids 13
pI 6.16, dissociation constants: pK 1 2.71, pK 2
9.62.
L-Thyroxine [51-48-9], 2-amino-3-[4-(4-
hydroxy-3,5-diiodophenoxy)-3,5-diiodophen-
yl]propionic acid, C15 H11 NO4 , M r 269.18, oc-
currence: thyroid gland.
D,L-Tryptophan [54-12-6], 2-amino-3-
(3 -indolyl)propionic acid, α-amino-1H-indole-
3-propanoic acid, C11 H12 N2 O2 , M r 204.23,
mp 285 ◦ C (decomp.), solubility 0.25 (30 ◦ C)
g/100 g H2 O, pI 5.88, dissociation constants:
pK 1 2.38, pK 2 9.39.
D,L-Tyrosine [556-03-6], 2-amino-3-
(4-hydroxyphenyl)propionic acid, [3-(4-hy-
droxyphenyl)]alanine, 2-amino-3-(p-hydroxy-
phenyl)propionic acid, α-amino-4-hydroxy-
benzenepropanoic acid, C9 H11 NO3 , M r 181.19,
mp 340 ◦ C, 318 ◦ C (decomp.), solubility 0.351
(25 ◦ C) g/100 g H2 O, pI 5.63, dissociation con-
stants: pK 1 2.2, pK 2 9.11, pK 3 10.07 (–OH).
D,L-Valine [516-06-3], 2-amino-3-
methylbutyric acid, 2-aminoisovaleric acid,
C5 H11 NO2 , M r 117.15, mp 298 ◦ C (decomp.,
sealed tube), solubility 7.09 (25 ◦ C) g/100 g
H2 O, pI 6.0, dissociation constants: pK 1 2.29,
pK 2 9.72.
D-Valine [640-68-6], 2-amino-3-methylbu-
tyric acid, d-Val, C5 H10 NO2 , M r 116.14, appli-
cation: Fluvalinate (insecticide) [51].
3. Industrial Production of Amino
Acids
Amino acids can be classified as natural, in-
cluding the proteinogenic (occurring in proteins)
amino acids, or as nonnatural. Originally, only
14 Amino Acids

Figure 2. Routes for production of amino acids

20 amino acids were believed to be genetically
coded, but the recent discoveries of selenocys-
teine and pyrrolysine have demonstrated that ad-
ditional amino acids could be coded by subvert-
ing a stop codon. The following sections will
focus on production methods for the principal
proteinogenic amino acids, which are of major
importance in nutrition and as feed additives.
This process is now generally being replaced by
other methods, because of customer preference
for materials not of animal origin. Some amino
acids, such as l-tyrosine, can be economically
isolated from plant residues, for example sugar
beet molasses.

3.1. General Methods

Four basic processes (see Fig. 2) are suitable for
the production of amino acids: chemical syn-
thesis (including asymmetric synthesis), extrac-
tion, fermentation, and enzymatic routes. The
classical chemical synthesis is applied to pro-
duce either the achiral amino acid glycine or
racemic amino acids, for instance d,l-methio-
nine. If l-amino acids are to be manufactured
by this route, the chemical synthesis has to be
followed by a resolution step. In some cases l-
amino acids are directly produced from prochiral
precursors by means of enzymes as chiral cata-
lysts. For l-cysteine, transformation of another
amino acid (cystine) is an important method.
Catalytic asymmetric syntheses have been de-
veloped for many l-amino acids and their d-
antipodes, but few have been applied on an in-
dustrial scale. The extraction process has the
advantage of offering access to nearly all the
proteinogenic l-amino acids by isolation from
protein hydrolysates. The starting materials are
protein-rich products, such as keratin, feathers,
blood meal, or technical gelatin (see Fig. 3).
Figure 3. Extraction of amino acids
 Amino Acids 15
Table 2. Production of amino acids, methods, and production
volume
are resistant to toxic compounds, i.e., analogues
Type Amino acid Preferred production or derivatives of amino acids. In such cases high
method amino acid concentrations do not inhibit the en-
I l-Alanine enzymatic catalysis, zymatic key in the pathway of biosynthesis and
fermentation
I l-Asparagine extraction, chemical thus result in high production rates of the de-
synthesis sired amino acid. The fourth method, enzymatic
I l-Glutamine fermentation, extraction catalysis, uses whole cells or active cell com-
I l-Histidine fermentation, extraction
I l-Hydroxyproline fermentation, extraction ponents (enzymes) as such or, if possible, as
I l-Isoleucine fermentation, extraction immobilized biocatalysts in continuously oper-
I l-Leucine fermentation, extraction ated reactors. The competitiveness of enzymatic
I l-Methionine enzymatic resolution
I l-Proline fermentation, extraction
processes depends on the availability and price
I l-Serine fermentation, extraction of substrate, the activity and stability of the in-
I l-Tyrosine extraction volved enzyme(s) and on the simplicity of prod-
I l-Valine enzymatic catalysis, uct recovery. For some general reviews on ami-
fermentation
II l-Arginine fermentation, extraction
no acid synthesis, emphasizing the chemical and
II l-Cysteine fermentation, enzymatic biocatalytic methods, see [52–56].
catalysis (via reduction of In Table 2 the amino acids and their preferred
l-cystine)
II l-Tryptophan fermentation
production methods are listed, classified by the
III Glycine chemical synthesis
size of production volume. Table 3 specifies the
III l-Aspartic acid enzymatic catalysis amino acids produced by direct fermentation of
III l-Phenylalanine fermentation carbohydrates.
III l-Threonine fermentation
IV d,l-Methionine chemical synthesis and
enzymatic resolution
IV l-Glutamic acid fermentation 3.2. Production of Specific Amino Acids
IV l-Lysine fermentation
Type I 100–1000 t/a 3.2.1. L-Alanine
Type II 1000–8000 t/a
Type III 8000–100 000 t/a
Type IV 100 000–800 000 t/a
l-Alanine is industrially produced from l-
aspartic acid by means of immobilized Pseu-
domonas dacunhae cells in a pressurized biore-
The use of overproducing microbial strains actor [69]. In direct fermentation microorgan-
(→ Biotechnology; → Enzymes, Chap. 6.5) in isms usually accumulate d,l-alanine because
fermentation processes with sucrose or glucose of alanine racemase also present. With a d-cy-
as carbon source is currently the most economic closerine resistant mutant selected from Bre-
production method for bulk amino acids such vibacterium lactofermentum, it is possible to ob-
as monosodium l-glutamate, l-lysine hydro- tain 46 g/L d-alanine with an enantiomeric ex-
chloride, and l-threonine. In the last ten years, cess (e.e.) of 95 % [58]. An alanine racemase-
this has become the method of choice for pro- deficient mutant of Arthrobacter oxydans was
duction of other amino acids, such as l-phenylal- reported, that produces 75 g/L l-alanine from
anine, l-tryptophan, and l-cysteine. Classical glucose with a yield of 52 % and 95 % e.e. [57].
breeding and mutagenesis as well as recombi- A small amount of l-alanine is still isolated from
nant DNA techniques are generally employed protein hydrolysates.
to modify the biosynthetic pathway and force
the bacteria to overproduce the corresponding
metabolites. Industrially used mutants are usu- 3.2.2. l-Arginine
ally characterized by genetic markers introduced
by methods of deregulation, e.g., auxotrophic L-Arginine is today produced mainly by fer-
mutants, in order to release key enzymes from mentation. Suitable strains for fermentation are
strict regulation by metabolites (feedback inhi- deregulated mutants derived from Corynebac-
bition and repression). A different way to obtain terium glutamicum and Bacillus subtilis ac-
overproducing strains consists of screening of cumulating 25–35 g/L l-arginine from glucose
regulatory mutants, in that important enzymes [70]. Very potent strains of Serratia marcescens
16 Amino Acids
Table 3. Amino acids produced by direct fermentation from carbohydrates
Amino acid Type of mutant Amino acid yield, g/L Reference
l-Alanine (95 % e.e.) Athrobacter oxydans 75 Hashimoto and Katsumata (1994)
[57]
d-Alanine (95 % e.e.) Brevibacterium lactofermentum 46 Yahata et al. (1993) [58]
l-Arginine Serratia marcescens AUr -1 100 Chibata et al. (1983) [59]
l-Glutamine Brevibacterium flavum AJ 3409 57 Yoshihara et al. (1992) [60]
l-Histidine Serratia marcescens 40 Sugiura et al. (1987) [61]
l-Isoleucine Escherichia coli H-8461 30 Kino et al. (1993) [62]
l-Leucine Brevibacterium flavum AJ 3686 19.5 Yoshihara et al. (1992) [63]
l-Lysine Corynebacterium glutamicum >120 Oh et al. (1990) [64]
l-Methionine Pseudomonas putida VKPM V-4167 3.5 University Odessa (1992) [65]
l-Phenylalanine Escherichia coli MWPWJ304/pMW16 51 Ikeda et al. (2003) [56]
l-Proline Brevibacterium flavum AP113 97.5 Kocarian et al. (1986) [66]
l-Serine Methylobacterium sp. MN43 65 Ikeda et al. (2003) [56]
l-Threonine Escherichia coli BKIIM B-3996 85 Debabov et al. (1990) [67]
l-Tryptophan Corynebacterium glutamicum 50 Ikeda et al. (1994) [68]
KY9218/pKW9901
l-Valine Corynebacterium glutamicum VR 3 99 Ikeda et al. (2003) [56]

derived from mutants obtained by transduc-
tion, having feedback-insensitive and derepres-
sive enzymes of arginine biosynthesis and 6-
azauracil resistance, are able to produce 60–
100 g/L l-arginine [59]. A small amount of l-
arginine is isolated from protein hydrolysates.
3.2.3. l-Aspartic Acid and Asparagine
L-Aspartic acid is industrially manufac-
tured by an enzymatic process in which as-
partase (l-aspartate ammonia lyase, EC 4.3.1.1)
catalyzes the addition of ammonia to fumaric
acid [71]. Advantages of the enzymatic produc-
tion method are higher product concentration
and productivity and the formation of fewer by-
products. Thus l-aspartic acid can be easily sep-
arated from the reaction mixture by crystalliza-
tion.
A process involving continuous production
of l-aspartic acid by means of carrier-fixed as-
partase isolated from Escherichia coli was first
commercialized in Japan [72]. An economically
attractive process for l-aspartic acid uses rest-
ing or dried cells with a high aspartase content,
e.g., a 4.5 % suspension of aspartase-containing
Brevibacterium flavum cells could be recycled in
seven repeated batches and concentrations up to
166 g/L l-aspartate could be achieved [73]. In
1973, an immobilized cell system based on E.
coli cells entrapped in polyacrylamide gel lat-
tice was introduced for large-scale production
[74]. This example represents the first industrial
application of immobilized microbial cells in a
fixed-bed reactor. Further improvements, immo-
bilization of the cells in κ-carrageenan, provided
remarkably increased operational stability, re-
sulting in biocatalyst half-lives of almost two
years [75]. A column packed with the κ-carra-
geenan-immobilized system allows a theoretical
productivity of 140 g L−1 h−1 l-aspartate.
As l-aspartic acid gained importance as in-
termediate for the manufacture of the dipep-
tide sweetener aspartame (methyl ester of l-
aspartyl-l-phenylalanine) improved processes
were developed. For continuous production
of l-aspartic acid Escherichia coli strains
immobilized with polyurethane [76] or with
polyethylenimine and glass fiber support [77]
and κ-carrageenan-immobilized Pseudomonas
putida [78] have been reported and protected,
respectively. Less successful was the search for
strains that are overproducing l-aspartate by fer-
mentation of sugar. A pyruvate kinase-deficient
mutant of Brevibacterium flavum accumulates
up to 22.6 g/L l-aspartic acid in glucose-con-
taining medium [79], not enough to be compet-
itive with the enzymatic processes. On the other
hand, genetic recombination techniques have
helped to improve aspartase-containing strains.
An aspA gene bearing plasmid (pBR322:aspA-
par) was able to elevate aspartase formation in
Escherichia coli K 12 about 30-fold [80].
L-Asparagine can be isolated as a byproduct
from the production of potato starch. A simple
synthesis of l-asparagine starts from l-aspartic
acid which is esterified to the β-methyl ester fol-
lowed by treatment with ammonia [81].

3.2.4. L-Cystine and L-Cysteine
l-Cysteine used to be produced almost exclu-
sively by hydrolysis of hair or other keratins. The
amino acid isolated was l-cystine, which was re-
duced electrolytically to l-cysteine. l-Cysteine
has also been prepared from β-chloro-d,l-ala-
nine and sodium sulfide with cysteine desulfhy-
drase, an enzyme obtained from, e.g., Citrobac-
terium freundii [82].
Today, however, the main processes for cys-
teine production are biological. A direct fer-
mentation process has been developed for the
manufacture of l-cystine, using a modified E.
coli bacterium [83]. The technology has been
extended to prepare other modified l-cysteine
analogues [84]. An enzymatic process for l-
cysteine has been successfully developed using
microorganisms capable to hydrolyze 2-amino-
∆2 -thiazoline 4-carboxylic acid (ATC) which is
readily available from methyl α-chloroacrylate
and thiourea. A mutant of Pseudomonas thi-
azolinophilum converts d,l-ATC to l-cysteine
in 95 % molar yield at product concentrations
higher than 30 g/L [85].
3.2.5. L-Glutamic Acid
In 1957, a soil bacterium was discovered [86]
which was able to excrete considerable amounts
of l-glutamate. Fermentation processes using
strains of this bacterium, later called Corynebac-
terium glutamicum, have been successfully
commercialized not only for L-glutamic acid
but also for the production of other economi-
cally important amino acids [87, 88]. Numer-
ous coryneform microorganisms have been iso-
lated and found to be able to overproduce l-
glutamic acid and other amino acids. Exam-
ples of these microorganisms are Brevibac-
terium flavum, Brevibacterium lactofermentum,
and Microbacterium ammoniaphilum. Because
of minor differences in the character of those
bacteria [89] which are all gram-positive, non-
spore-forming, nonmotile and all require biotin
for growth, the name of genus Corynebacterium
was suggested for these coryneform bacteria
[90].
For industrial production of l-glutamic acid,
molasses (sucrose), starch hydrolysates (glu-
cose) and ammonium sulfate are generally used
Amino Acids 17
as carbon and nitrogen sources, respectively
[91]. Key factors in controlling the fermentation
are the presence of biotin in optimal concentra-
tion — to optimize cell growth and the excretion
of l-glutamate — and sufficient supply of oxy-
gen to reduce the accumulation of byproducts,
such as lactic and succinic acid. In biotin-rich
fermentation media the addition of penicillin or
cephalosporin C favors the overproduction of l-
glutamic acid due to effects on the cell mem-
brane. The supplementation of fatty acids also
results in an increased permeability of the cells
thus enhancing glutamate excretion. In the past
the mechanism of glutamate excretion was sim-
ply explained as a “leakage” or “overflow” phe-
nomenon [92]. In the beginning of the 1990s it
was reported that a specific carrier system exists
[93, 94], which is responsible for active gluta-
mate transport in Corynebacterium glutamicum.
Generally the intracellular accumulation of glu-
tamate does not reach levels sufficient for feed-
back control in glutamate overproducers due to
rapid excretion of glutamate. However, the reg-
ulatory mechanisms of l-glutamic acid biosyn-
thesis have been studied intensively to obtain
mutants with increased productivity. Two en-
zymes have been shown to play key roles in the
biosynthesis of l-glutamic acid [95].
1) Phosphoenolpyruvate carboxylase (PEPC)
catalyzes carboxylation of phosphoenolpyru-
vate to yield oxaloacetate; it is inhibited by l-
aspartic acid and repressed by both l-aspartic
and l-glutamic acids.
2) α-Ketoglutarate dehydrogenase (KDH) con-
verts α-ketoglutarate to succinyl-CoA. In l-
glutamate overproducing strains KDH limits
further oxidation of α-ketoglutarate to carbon
dioxide and succinate, thus favoring forma-
tion of l-glutamic acid.
In l-glutamate overproducing strains the K m
value of KDH for α-ketoglutarate was nearly
two magnitudes lower than that of l-glutamic
acid dehydrogenase (GDH) which catalyzes
the last step, the reductive amination of α-
ketoglutarate to l-glutamate. Consequently,
V max of GDH was proven to be about 150 times
higher than that of KDH. The Corynebacterium
glutamicum gdh gene has been isolated and
characterized [96]. A strain of Microbacterium
ammoniaphilum cultured under biotin-deficient
conditions produced 58 % of l-glutamic acid
18 Amino Acids

formed from glucose via phosphoenolpyruvate, product is started by biomass separation using
citrate, and of α-ketoglutarate and the other 42 % centrifuges or ultrafiltration units, concentration
via the tricarboxylic acid (TCA) or the glyoxy- of the centrifugate or filtrate, followed by crys-
late cycle [97]. In large-scale production the for- tallization, filtration and drying. A scheme of
mation of trehalose very often reduces the prod- production steps is given in Figure 4.
uct yield. Trehalose consists of two α-1,1 bound
glucose molecules and is excreted by the bacte-
ria as a material with protects the cell against
high osmotic pressure (osmoprotectant). A pro-
cess was recently developed and successfully in-
dustrialized in which trehalose formation is con-
trolled and decreased by culturing the overpro-
ducing mutant in media containing invert sugar
from molasses [98, 99].
Today wild type isolates as well as mutants
developed by classical breeding or even strains
constructed by modern techniques, using cell fu-
sion or recombinant DNA methods [100], are
available for industrial l-glutamate production.
In addition a new type of fermentation process
was reported which uses a strain that overpro-
duces l-glutamic acid and l-lysine simultane-
ously. The cultivation of an auxotrophic regula-
tory mutant of Brevibacterium lactofermentum
in a medium, supplemented with polyoxyeth-
ylenesorbitan monopalmitate as surface-active
agent, resulted in the accumulation of 162 g l-
amino acids per liter (105 g l-lysine · HCl + 57 g
l-glutamic acid). This corresponds to a produc-
tion rate of 3.8 g (l-lysine · HCl + l-glutamic
acid) per liter per hour [101].
The success of an economic production ba-
sically depends on the experience in fermenta-
tion technology, in up- and downstream pro-
cessing and on the skill of employees in re-
search, development, and production. A multi-
step inoculation procedure followed by the fed-
batch mode (→ Biotechnology, Chap. 6.2) in the
main fermentation up to 500 m3 scale is still the
preferred technology for the production of l- Figure 4. Flow diagram of fermentation and downstream
processing of l-glutamic acid
glutamic acid.
Critical operations are the steam-forced ster-
ilization of the fermenter and batchwise or
continuous sterilization of the culture medium 3.2.6. L-Glutamine
to prevent contamination by foreign microbes.
During fermentation the temperature, pH, dis- Microbial l-glutamine producers were selected
solved oxygen, and the sugar consumption have from wild type glutamate-producing coryne-
to be controlled as important process parame- form bacteria. A sulfaguanidine resistant mutant
ters. When the fermentation is completed after of Brevibacterium flavum accumulates 41 g/L l-
40–60 h, the production strain may have accu- glutamine in 48 h from 10 % glucose [102]. A
mulated more than 150 g/L l-glutamic acid. Af- yield of 44 % was achieved by the mutant Bre-
ter deactivation of the broth, the recovery of the vibacterium flavum AJ3409 [60].

3.2.7. L-Histidine
Efficient l-histidine fermentation can be per-
formed with strains of Corynebacterium glu-
tamicum and Serratia marcescens. A mutant
of Corynebacterium glutamicum having re-
sistance to 8-azaguanine, 1,2,4-triazole-3-ala-
nine, 6-mercaptoguanine, 6-methylpurine, 5-
methyltryptophan and 2-thiouracil, produces 15
g/L of l-histidine. Only after introduction of
these resistance markers by mutagenesis is the
mutant capable of releasing this amount of l-
histidine into the nutrient solution. A strain
of Serratia marcescens having both characters,
feedback-insensitive and derepressed histidine
enzymes combined with transductional tech-
niques and 6-methylpurine resistance, accumu-
lates 23 g/L l-histidine [103]. By amplification
of the genes hisG, hisD, hisB, hisC in Ser-
ratia marcescens the final concentration of l-
histidine could be elevated from 28 g/L to 40 g/L
[61].
l-Histidine can also be produced by isolation
from blood-meal hydrolysates [104].
3.2.8. L-Hydroxyproline
l-Hydroxyproline (trans-4-hydroxy-l-proline,
2S,4R-hydroxyproline) is today mainly obtained
by enzymatic hydroxylation of l-proline with
genetically modified E. coli [105, 106]. The
proline 4-hydroxylase specifically produces the
4-trans isomer. This method has almost com-
pletely replaced the isolation from hydrolysates
of gelatin or other collagens. An unusual fea-
ture of this amino acid is that it is not incorpo-
rated into collagen during biosynthesis at the ri-
bosome, but is formed from l-proline by a post-
translational hydroxylation reaction.
3.2.9. L-Isoleucine
An advantageous fermentation method for pro-
duction of l-isoleucine is the use of chem-
ically synthesized substrates that only re-
quire a few steps to be converted to l-
isoleucine. Among these the natural precursors
2-ketobutyrate [107] or d,l-2-hydroxybutyrate
[108] have been used for the production with
Amino Acids 19
Corynebacterium glutamicum. A leucine re-
quiring mutant of Corynebacterium glutam-
icum, with increased d-lactate utilization and
consuming d,l-2-hydroxybutyrate, accumulates
13.4 g/L l-isoleucine. However, exploitation of
this process is hampered by formation of by-
products [109]. Sugar based l-isoleucine pro-
cesses have been developed with strains of
Corynebacterium glutamicum [110], Serratia
marcescens [111], and Escherichia coli [112–
114]. The mutant Escherichia coli H-8285, be-
ing resistant to thiaisoleucine, arginine hydrox-
amate, and d,l-ethionine accumulates 26 g/L l-
isoleucine in 45 h in a fed-batch process [115].
Introduction of resistance to 6-dimethylami-
nopurine in strain H-8285 resulted in a mu-
tant Escherichia coli H-8461 that increased
l-isoleucine accumulation to 30.2 g/L [62].
The biosynthesis of l-isoleucine has been in-
vestigated in detail on the level of involved
genes [63], thus recombinant strains are be-
ing constructed with high productivity and se-
lectivity. An appropriate balance of homoser-
ine dehydrogenase and threonine dehydratase
activities in the construct Corynebacterium
glutamicum DR17/pECM3::ilvA(V323A) with
feedback-resistant aspartate kinase create a spe-
cific productivity of 0.052 g l-isoleucine per
gram dry biomass per hour [116]. The recombi-
nant strain Escherichia coli AJ13100 produces
l-isoleucine from glucose with high selectivity
in 30 % yield [117].
3.2.10. L-Leucine
l-Leucine can be manufactured by fermentation,
precursor fermentation, or, less commonly to-
day, by isolation from protein hydrolysates. Us-
ing a fed-batch culture of Corynebacterium glu-
tamicum ATCC 13032 32 g/L 2-ketoisocaproate
can be converted to 24 g/L l-leucine by the
transaminase B reaction [118]. With the method
of direct fermentation l-leucine can be produced
by either α-aminobutyric acid-resistant mutants
of Serratia marcescens or by 2-thiazole-alanine-
resistant coryneform strains [119]. Brevibac-
terium flavum AJ3686 accumulates 19.5 g/L l-
leucine (15 % yield) [120].
20 Amino Acids
3.2.11. L-Lysine
The most potent microorganisms to overproduce
l-lysine are mutants derived from Corynebac-
terium glutamicum, a gram-positive bacterium
first introduced as an l-glutamate producing mi-
crobe; the wild strains themselves are not able
to excrete l-lysine. Mainly auxotrophic and reg-
ulatory mutants of this bacterium have been de-
veloped by classical breeding methods and mu-
tagenesis. The following techniques have been
applied to channel the metabolic pathway in the
biosynthesis of amino acids:
Screening for auxotrophic mutants, in order
to release key enzymes, for instance aspartate
kinase, from strict regulation by metabolites
(feedback inhibition).
Screening for regulatory mutants, in which as-
partate kinase is resistant to toxic analogues of
l-lysine, such as S-(2-aminoethyl)-l-cysteine
(AEC) or O-(2-aminoethyl)-l-serine. In such
strains high lysine concentrations do not in-
hibit the enzymatic key step, the formation
of 4-aspartylphosphate from l-aspartase cat-
alyzed by aspartate kinase.
Screening for mutants having amino acid aux-
otrophy combined with deregulation.
Screening for regulatory mutants having ad-
ditional enzyme defects (reduced pyruvate ki-
nase) and low levels of other enzymes (citrate
synthase).
As a modern technique, cell fusion with the
method of protoplast fusion has been succes-
sively applied for breeding of industrial microor-
ganisms [121, 122]. This technique allows the
combination of positive characteristics of dif-
ferent strains such as high selectivity and high
productivity. In fermentation with media of in-
hibitory osmotic stress (i.e., high substrate con-
centration results in high osmotic pressure on
the cell that inhibits the performance of the mu-
tant) the sugar consumption rate and l-lysine
production rate of some mutants can be stimu-
lated by the addition of glycine [123]. Another
attractive approach is the development of ther-
mophilic strains. A mutant of Corynebacterium
thermoaminogenes was patented which is capa-
ble of growing at a temperature > 40 ◦ C and
accumulating l-lysine in the culture medium
[124].
Meanwhile, regulation of lysine excretion in
overproducing strains is known in detail [125,
126]. Thus more attention should be paid to pro-
cess development in the fields of molecular biol-
ogy, biochemistry, and physiology and to finding
new approaches for developing improved over-
producing mutants [127]. The most specific and
well-directed methods for strain development
are offered by recombinant DNA techniques.
In principle all genes encoding the relevant
enzymes in l-lysine biosynthesis have been iso-
lated, characterized, and amplified in coryne-
form bacteria to enhance l-lysine formation
[128]. As an example, amplification of dapA
gene that codes for dihydrodipicolinate syn-
thase in Corynebacterium glutamicum resulted
in 35 % higher overproduction of l-lysine com-
pared to the parent strain [129]. Another option
for strain improvement is the transformation of
dapA gene together with a lysC gene, coding for
aspartate kinase with decreased feed back inhi-
bition in Corynebacterium glutamicum [130].
In fed-batch culture and under appropriate
conditions the favorable mutants for lysine pro-
duction are able to reach final concentration
of about 120 g/L l-lysine, calculated as hydro-
chloride [64]. Fermentation processes are per-
formed in big tanks up to 500 m3 size. An op-
timized feeding strategy practiced with a com-
puter aided process control system may enable
high conversion yield and productivity in large
scale fed-batch cultivation [131]. Apart from
the specific fermentation know how, inoculation,
sterilization, and feeding strategy the recovery
process and the quality of the product both can
be decisive factors to guarantee competitiveness.
The conventional route of lysine downstream
processing is characterized by:
Removal of the bacterial cells from fermenta-
tion broth by separation or ultrafiltration
Absorbing and then collecting lysine in an ion
exchange step
Crystallizing or spray drying of lysine as l-
lysine hydrochloride
An alternative process consists of biomass sep-
aration, concentration of the fermentation solu-
tion, and filtration of precipitated salts. The liq-
uid product contains up to 50 % l-lysine base,
that is stable enough to be marketed [132]. In the
1990’s, a new concept for lysine production was
introduced. Here the lysine containing fermen-
tation broth is immediately evaporated, spray-

dried, and granulated to yield a feed-grade prod-
uct, which contains lysine sulfate. Its lysine con-
tent correspond to that of a material which con-
tains to at least 60 % of l-lysine hydrochloride
[133, 134]. The process avoids any waste prod-
ucts usually present in the conventional l-lysine
hydrochloride manufacture.
3.2.12. D,L-Methionine and L-Methionine
l-Methionine and its antipode d-methionine are
of equal nutritive value, thus the racemate can
directly be used as feed additive.
The most economic way for production of
d,l-methionine is the chemical process based on
acrolein, methyl mercaptan, hydrogen cyanide,
and ammonium carbonate (see Fig. 5). β-
Methylthiopropionaldehyde, formed by addi-
tion of methyl mercaptan to acrolein, is the in-
termediate that reacts with hydrogen cyanide
to give α-hydroxy-γ-methylthiobutyronitrile.
Treatment with ammonium carbonate leads to
5-(β-methylthioethyl)hydantoin that is saponi-
fied by potassium carbonate giving d,l-methio-
nine in up to 95 % yield, calculated on acrolein
[135].
Figure 5. Degussa process for production of l-methionine
Amino Acids 21
The production method of choice for L-
methionine is still the enzymatic resolution
of racemic N-acetyl-methionine using acylase
from Aspergillus oryzae. The production is car-
ried out in a continuously operated fixed-bed or
enzyme membrane reactor [136].
Alternatively, l-methionine may be produced
by microbial conversion of the correspond-
ing 5-substituted hydantoin. With growing cells
of Pseudomonas sp. strain NS671, d,l-5-(2-
methylthioethyl)hydantoin was converted to l-
methionine; a final concentration of 34 g/L and
a molar yield of 93 % have been obtained [137].
Biosynthesis of l-methionine and its regula-
tion in bacteria is well known. Although some
promising concepts, for example utilization of
sulfate, sulfite, or thiosulfate as sulfur sources
for microbes have been suggested [138], it was
not possible so far to develop strains that are able
to excrete remarkable amounts of l-methionine
into the culture medium.
3.2.13. L-Phenylalanine
Several chemical, biocatalytic, and fermentation
methods for producing l-phenylalanine have
been developed, some of which are of industrial
significance (see Fig. 6).
In previous large-scale production processes
for l-phenylalanine two enzymatic methods
were applied:
1) Resolution of N-acetyl-d,l-phenylalanine by
carrier-fixed microbial acylase: This process
provided pharmaceutical-grade l-phenylala-
nine, but suffered from the disadvantage that
the d-enantiomer had to be racemized and re-
cycled.
2) Stereoselective and enantioselective addition
of ammonia to trans-cinnamic acid, catalyzed
by l-phenylalanine ammonia lyase (PAL,
EC 4.3.1.5): PAL-containing Rhodotorula
rubra was used in an industrial process [139]
to supply l-phenylalanine for the first pro-
duction campaign of the sweetener aspar-
tame. When continuously operated in an im-
mobilized whole cell reactor, the biocon-
version reached concentration up to 50 g/L
l-phenylalanine at a conversion of about
83 % [140]. Other processes started from
phenylpyruvate with l-aspartic acid as amine
donor using immobilized cells of Escherichia
22 Amino Acids

Figure 6. Asymmetric syntheses for production of l-phenylalanine

coli [141] or from α-acetamidocinnamic acid a medium containing 13 % glucose. Similar re-
and immobilized cells of a Corynebacterium sults can be obtained by tyrosine auxotrophic
equi strain [142]. In both cases l-phenylal- regulatory mutants of E. coli [145]. With re-
anine concentrations up to 30 g/L and more combinant DNA techniques it was possible to
(molar yields as high as at least 98 %) were improve overproducing strains of coryneform
reached. bacteria as well as of E. coli. Amplification
of a deregulated DAHPS gene was achieved
However, fermentation processes based on
in a phenylalanine producer of Brevibacterium
glucose-consuming l-phenylalanine overpro-
lactofermentum [146]. For optimal production
ducing mutants of E. coli and coryneform strains
of l-phenylalanine in fed-batch cultivation the
turned out to be more economical. The biosyn-
critical specific glucose uptake rate has to be
thetic pathway for aromatic amino acids in bac-
controlled. The specific feed rate during fer-
teria is strictly regulated [143]. l-Phenylalanine
mentation has to be adjusted below a critical
is formed in ten enzymatic steps starting from
limit, since otherwise the E. coli producer will be
erythrose-4-phosphate and phosphoenolpyru-
forced to excrete acetate [147]. A suitable profile
vate. The biosynthesis is governed by the first
of the specific glucose feed rate prevents acetate
key enzyme 3-deoxy-d-arabinoheptulosonate-
formation and leads to improved l-phenylala-
7-phosphate synthase (DAHPS) which is in-
nine production with a final concentration up
hibited by both l-phenylalanine and l-tyrosine
to 46 g/L and a corresponding yield of 18 %. l-
(by acting on the enzyme itself) and repressed
Phenylalanine is recovered from the fermenta-
by l-tyrosine (by acting on the according gene
tion broth either by two-step crystallization or
of the DNA). The other important enzyme is
by an ion-exchange resin process. The preferred
prephenate dehydratase (PDT) also inhibited by
cell separation technique is ultrafiltration; and
l-phenylalanine, but stimulated by l-tyrosine.
the filtrates may be treated with activated carbon
To overcome these regulatory mechanisms ei-
for further purification. Instead of ion-exchange
ther auxotrophs of Corynebacterium glutam-
resins nonpolar, highly porous synthetic adsor-
icum have been constructed or l-phenylalanine
bents are recommended to remove impurities
analogues, e.g., 4-aminophenylalanine and 4-
[148, 149]. An alternative process in which a
fluorophenylalanine, have been applied. The lat-
cell separator is integrated in the fermentation
ter variant leads to resistant mutants of Brevibac-
pa rt, thus allowing cell recycling, was suggested
terium flavum or lactofermentum [144]. These
for l-phenylalanine production and may lead to
auxotrophic and regulatory mutants are able to
prospective developments [150].
produce more than 20 g/L of l-phenylalanine in

3.2.14. L-Proline
l-Proline is still produced to a small ex-
tent by isolation from protein hydrolysates,
but today direct fermentation using analogue-
resistant mutants of coryneform bacteria or
Serratia marcescens is an economic alterna-
tive production method [151]. An isoleucine
auxotrophic mutant of Brevibacterium flavum
having resistance to sulfaguanidine and d,l-
3,4-dehydroproline (DP) is able to accumulate
40 g/L l-proline. Brevibacterium flavum AP113
is claimed to produce 97.5 g/L l-proline; this
mutant is characterized by isoleucine auxotro-
phy, resistance to DP, and osmotic pressure
and incapable to degrade l-proline [66]. A pro-
line oxidase-less strain of Serratia marcescens,
having resistance to DP, thiazoline-4-carboxyl-
ate and azetidine-2-carboxylate, overproduces
58.5 g/L l-proline into the culture medium
[152]. By amplification of the genes proA and
proB in this type of regulatory mutant, a con-
struct was obtained which yields 75 g/L l-
proline [153].
3.2.15. L-Serine
l-Serine is obtained by microbial/ enzymatic
conversion of glycine using immobilized rest-
ing cells or crude cell extracts. Hyphomicro-
bium strains possess the serine pathway and
are able to produce l-serine from methanol
and glycine. Methanol is oxidized by methanol
dehydrogenase to formaldehyde which in turn is
converted in an aldol-like reaction with glycine
to l-serine. The reaction is catalyzed by ser-
ine hydroxymethyltransferase (SHMT) [154].
Hyphomicrobium sp. NCIB10099 was found to
produce 45 g/L l-serine from 100 g/L glycine
and 88 g/L methanol in three days [155]. In
an enzyme bioreactor with a feedback control
system a crude extract from Klebsiella aero-
genes containing SHMT has been used to syn-
thesize l-serine from glycine and formaldehyde
in the presence of tetrahydrofolic acid and pyri-
doxal phosphate. In this bioreactor a serine con-
centration of 450 g/L with an 88 % molar con-
version of glycine at a volumetric productiv-
ity of 8.9 g L−1 h−1 could be achieved under
optimized conditions [156]. With whole cells
of Escherichia coli MT-10350 l-serine is pro-
duced by treatment of an oxygenated aqueous
Amino Acids 23
glycine solution (485 g/L) with aqueous form-
aldehyde for 35 h at 50 ◦ C in a molar yield of
89 % based on glycine [157]. Extraction of pro-
tein hydrolysates is used to a smaller extent.
3.2.16. L-Threonine
Up to the end of the 1980s, l-threonine was
mainly used for medical purposes, in amino acid
infusion solutions and nutrients. It was manu-
factured by extraction of protein hydrolysates
or by fermentation using mutants of coryneform
bacteria in amounts of several hundred tons per
year worldwide. The production strains were de-
veloped by classical breeding. They were aux-
otrophic and resistant to threonine analogues
such as α-amino-β-hydroxyvalerate (AHV),
and reached product concentrations up to 20 g/L.
These strains possessed deregulated l-threonine
pathways with feedback inhibition-insensitive
aspartate kinase and homoserine dehydrogenase
[158, 159]. In the 1990s strain developments, us-
ing both conventional methods and recombinant
DNA techniques, have been very successful.
Potent classically selected mutants suggested
for industrial production are the species Bre-
vibacterium flavum, Providentia rettgeri, Serra-
tia marcescens, and Escherichia coli. However,
in the competition between the favorable can-
didates, strains of Escherichia coli proved to be
superior to other bacteria. Although the pathway
of l-threonine biosynthesis in Escherichia coli
is much more regulated than that in Corynebac-
terium glutamicum, new Escherichia coli strains
with excellent yields and productivity in threo-
nine formation could be constructed by genetic
engineering. l-Threonine is successfully mar-
keted as feed additive with a worldwide demand
of more than 10 000 t/a. Production strains are
based on Escherichia coli K-12 constructs har-
boring plasmids containing the thr operon that
consists of the genes thrA, thrB, and thrC [160].
Further improvements resulted in strains capa-
ble to accumulate more than 80 g/L in about
30 h with a conversion yield of more than 40 %
[161]. The strain stability could be further im-
proved, for example by integrating the threonine
operon into the chromosome [162]. The recov-
ery of feed-grade l-threonine is rather simple.
After fermentation is completed, cell mass is re-
moved by centrifugation or ultrafiltration, the
24 Amino Acids
filtrate is concentrated, depigmentated and l-
threonine isolated by crystallization [163].
3.2.17. L-Tryptophan
l-Tryptophan is one of the limiting essential
amino acids required in the diet of pig and poul-
try. A mature and growing market for l-trypto-
phan as feed additive is in development,based on
improved microbiological processes, and pro-
duction has increased to several thousand tons
per year. The most attractive production pro-
cesses for tryptophan are based on microorgan-
isms used as enzyme sources or as overproduc-
ers:
Enzymatic production from various precur-
sors
Fermentative production from precursors
Direct fermentative production from carbohy-
drates by auxotrophic and analogue resistant
regulatory mutants
l-tryptophan is synthesized from indole, pyru-
vate, and ammonia by the enzyme tryptophanase
[164] or from indole and l-serine/d,l-serine
by tryptophan synthase [165, 166]. Although
production in enzyme bioreactors is quite effi-
cient and concentrations of l-tryptophan up to
200 g/L could be achieved by condensation of
indole and l-serine [167], these process vari-
ants were not economic due to the high costs
of the starting materials. The microbial con-
version of biosynthetic intermediates such as
indole or anthranilic acid to l-tryptophan has
also been considered as alternative for pro-
duction. Whereas indole consuming mutants of
Corynebacterium glutamicum produced about
10 g/L l-tryptophan [168], strains of Bacillus
subtilis and Bacillus amyloliquefaciens reached
final concentrations > 40 g/L l-tryptophan with
anthranilic acid as carbon source [168, 169].
The process with anthranilic acid as precursor
has been commercialized in Japan. However, the
manufacturer using genetically modified strains
derived from Bacillus amyloliquefaciens IAM
1521 was forced to stop l-tryptophan produc-
tion. l-Tryptophan produced by this process was
stigmatized because of side products found in
the product causing a new severe disease termed
eosinophilia-myalgia syndrome (EMS) [170].
One of the problematic impurities, “Peak E”,
was identified as 1,1 -ethylidene-bis-(l-trypto-
phan), a product formed by condensation of one
molecule acetaldehyde with two molecules of
tryptophan [171].
In other processes, i. e., direct fermenta-
tion using overproducing mutants and carbo-
hydrates as carbon sources, formation of such
impurities does not occur. In the 1990s strik-
ing progress has been made in the development
of auxotrophic and deregulated mutants of Bre-
vibacterium flavum, Corynebacterium glutam-
icum, and Bacillus subtilis. The biosynthesis of
L-tryptophan and its regulation have been re-
viewed in detail for the different species [172–
174]. The precise knowledge about the struc-
ture of the trp operon in Escherichia coli com-
prising the trp promoter and the genes trpE,
trpD, trpC, trpB, and trpA which are coding
for the enzymes anthranilate synthase (AS),
phosphoribosyl anthranilate transferase (PRT),
indole glycerol phosphate synthase (IGP) and
tryptophan synthase (TS), respectively, was the
benefit for further strain improvements.
Thus recombinant DNA techniques have
been used to increase the capability of overpro-
duction especially in strains of Corynebacterium
glutamicum and Escherichia coli. One concept
was realized successfully by amplification of trp
operon genes together with serA which codes
for phosphoglycerate dehydrogenase. This key
enzyme in l-serine biosynthesis should provide
enough l-serine in the last step of l-trypto-
phan formation. Production strains are able to
accumulate 30–50 g/L l-tryptophan with yields
higher than 20 % based on carbohydrate. Isola-
tion of l-tryptophan is still a major issue, be-
cause the amino acid is sensitive to oxygen and
heat [167]. However, at a typical production con-
centration of 50 g/L, more than half of the l-tryp-
tophan product crystallizes from the medium
[68, 175].
3.2.18. L-Tyrosine
l-Tyrosine was, until recently, produced exclu-
sively from protein hydrolysates. Its low solubil-
ity in water enables a quite simple isolation of
the amino acid. Enzymatic catalysis using a b-
tyrosinase (tyrosine-phenol lyase) from Erwinia
herbicola gives tyrosine by a three-component
synthesis from phenol, pyruvate, and ammonia.

This process has been commercialized for the
enzymatic synthesis of the drug l-DOPA, where
phenol is replaced by catechol [54]. l-Tyrosine
is formed in nature as part of the shikimic acid
pathway, as are l-phenylalanine and l-trypto-
phan. Recombinant DNA technology has been
used to develop strains of Corynebacterium glu-
tamicum that are specific for l-tyrosine, and pro-
duction yields of 20–25 g/L have been obtained
[175].
3.2.19. L-Valine
l-Valine is produced industrially in pharma-
ceutical quality by enzymatic resolution of
N-acetyl-d,l-valine. Using direct fermenta-
tion the branched-chained amino l-valine can
be produced by either α-aminobutyric acid-
resistant mutants of Serratia marcescens or by
2-thiazolealanine-resistant coryneform strains
[176]. Brevibacterium lactofermentum AJ12341
produces 39 g/L l-valine (28 % yield) [177].
4. Biochemical and Physiological
Significance
The biosynthesis of amino acids begins with at-
mospheric nitrogen, which is reduced to ammo-
nia by bacteria and plants. Ammonia is used by
plants, by bacteria, and, to a limited extent, by
ruminants as a raw material for amino acids.
Amino acids, in turn, serve as starting materi-
als for the synthesis of proteins and a variety
of other nitrogen-containing compounds, such
as the purine and pyrimidine bases in nucleic
acids. Bacterial degradation leads, once again,
to ammonia and nitrogen [178].
Figure 7. The transamination of amino acids
Amino Acids 25
α-Ketoglutaric acid plays the central role in
the assimilation of ammonia. Its transamination
product, glutamic acid, in turn, can provide its
amino group for the synthesis of other amino
acids, e.g., alanine (Fig. 7).
Humans, animals, and some bacteria are in-
capable of synthesizing all the necessary ami-
no acids in their own intermediary metabolism;
i.e., they are heterotrophs and are therefore de-
pendent on the biosynthetic capability of plants.
Proteins that are consumed as foods by humans
and animals are hydrolyzed to amino acids by
the digestive enzymes. The amino acids are re-
sorbed in the upper part of the intestine and enter
the liver by way of the portal vein. The liver is the
central organ for metabolism and homoeostasis
of the plasma amino-acid level. The body’s var-
ious requirements are met from the pool of free
amino acids (Fig. 8), ca. 50 g in adult humans.
Figure 8. The amino acid pool and functions of amino
acids in the intermediary metabolism
The lion’s share of the amino acids (≈ 300 g/d
for adults) is required for synthesis of proteins
[179]: structural proteins, enzymes, transport
proteins, and immune proteins. Additionally,
amino acids are required for the synthesis of
oligopeptides and polypeptides that fulfill reg-
ulatory functions in the body, i.e., hormones.
26 Amino Acids
Some amino acids or their metabolites are di-
rectly active as hormones or facilitate the trans-
mission of nerve impulses (neurotransmitters),
e.g., serotonin. Furthermore, there are amino
acids that serve special functions, such as methi-
onine, which is a methyl group donor. Finally,
a series of amino acids serves as precursors for
the biosynthesis of other structures. For exam-
ple, glycine is used in the construction of the por-
phyrin skeleton. Amino acids are metabolized to
produce energy in the case of a protein-deficient
or a protein-excess diet.
The free amino acids in the amino acid pool
undergo numerous transformations (Fig. 9), in-
volving transamination and oxidative deamina-
tion, by which other amino acids can be syn-
thesized. The α-keto acids are intermediates
and in addition allow amino acids entrance into
the carbohydrate (through pyruvate) and fatty
acid (through acetylcoenzyme A) metabolisms.
A distinction is therefore drawn between gluco-
genic and ketogenic amino acids.
Figure 9. Intermediary metabolism of amino acids
(simplified)
d-Amino acids occur in the cell pool of
plants and gram-positive bacteria and as build-
ing blocks in peptide antibiotics and bacterial
cell walls [10, 14]. They do not occur in human
or animal metabolism; proteins are made exclu-
sively of l-amino acids. The traces of d-ami-
no acids detected in metabolically inert protein
(teeth, eye lenses) are believed to originate from
racemization. Orally ingested d-amino acids are
resorbed from the intestinal lumen more slowly
than the l-form. The d-enantiomers cannot be
utilized or can be utilized only to a slight ex-
tent as essential amino acids [180]. The one
important exception is d-methionine. Animals
and adult humans convert d-methionine into l-
methionine by transamination. The α-keto acid
of methionine is an intermediate. Otherwise, d-
amino acids are degraded with the help of d-ami-
no acid oxidases [181] to be used as an energy
source [182].
The major end products of amino acid
metabolism are urea, uric acid, ammonium salts,
creatinine, and allantoin. The loss of nitrogen via
these metabolites stabilizes at about 22 g protein
per day after a few days on a protein-free diet.
Inborn disorders in amino acid metabolism
[183] can lead to marked alterations in the ex-
cretion profile. These disorders usually take the
form of an enzyme or transport deficiency [178,
184]. The most common example is phenylke-
tonuria, a disruption of the normal metabolic
pathway from phenylalanine to tyrosine caused
by severe limitation in the activity of the
phenylalanine hydroxylase [185].
Humans and animals are not capable of pro-
ducing all the required l-amino acids in their in-
termediary metabolism. Therefore, they are de-
pendent on an external source of these essential
amino acids (Table 4). In situations of increased
requirements (rapid growth, stress, trauma), his-
tidine and arginine also become essential for hu-
mans. Cysteine and tyrosine may be essential for
infants during their first few weeks, because their
intermediary metabolism does not yet function
well enough to produce these from methionine
and phenylalanine in sufficient quantities.
5. Uses
The uses of amino acids have been treated in
review articles [19, 186–189].
5.1. Human Nutrition
In addition to their nutritive value, amino acids
are important flavor precursors and taste en-
hancers. In foods for humans, the flavor uses
of amino acids represent the dominant factor in
 Amino Acids 27
Table 4. Essential (+) and semiessential (±) amino acids
Baby Adult Rat Chicken Hen Cat Salmon
l-Arginine ± ± ± + ± + +
l-Cysteine + (?)
Glycine +
l-Histidine + ± + + ± ± +
l-Isoleucine + + + + + + +
l-Leucine + + + + + + +
l-Lysine + + + + + + +
l-Methionine + + + + + + +
l-Phenylalanine + + + + + + +
l-Threonine + + + + + + +
l-Tryptophan + + + + + + +
l-Tyrosine + (?)
l-Valine + + + + + + +

Table 5. Average amino acid content of some foodstuffs (mg/100 g) a [190]

Food Ile Leu Lys Cyss Met Phe Tyr Thr Trp Val Arg His Protein, Moisture
% content,
%
Maize, grain 350 1190 254 147 182 464 363 342 67 M 461 398 258 9.5 12.0
Rice, husked 300 648 299 84 183 406 275 307 98 M 433 650 197 7.5 13.0
Wheat, 426 871 374 332 196 589 391 382 142 M 577 602 299 12.2 12.0
whole grain
Wheat, flour, 435 840 248 304 174 581 277 321 128 M 493 422 248 10.9 12.0
70–80 %
extr. rate
Potato 76 121 96 12 26 80 55 75 33 M 93 100 30 2.0 78.0
(Solanum
tuberosum)
Bean 927 1685 1593 188 234 1154 559 878 223 M 1016 1257 627 22.1 11.0
(Phaseolus
vulgaris)
Soybean, 171 278 195 57 50 175 133 128 48 M 165 253 84 3.2 92.0
milk
Soy protein, 4147 7119 5777 1008 1092 4644 3458 3211 1080 4210 6767 2378 75.7 4.7
isolate [191]
M
Lettuce, 50 83 50 24 – 67 35 54 10 71 59 21 1.3 94.8
leaves
(Lactuca
sativa)
Tomato 20 30 32 7 7 20 14 25 9M 24 24 17 1.1 93.8
(Solanum ly-
copersicum)
Apple 13 23 22 5 3 10 6 14 3 15 10 7 0.4 84.0
(Malus
silvestris)
Orange 23 22 43 10 12 30 17 12 6 31 52 12 0.8 87.4
(Citrus
sinensis)
Beef, veal, 852 1435 1573 226 478 778 637 812 198 M 886 1118 603 17.7 61.0
edible flesh
M
Fish, fresh, 900 1445 1713 220 539 737 689 861 211 1150 1066 665 18.8 74.1
all types
Milk, cows, 162 328 268 28 86 185 163 153 48 M 199 113 92 3.5 87.3
untreated
Milk, human 48 104 81 16 19 41 39 53 20 M 54 46 30 1.2 87.6
Cheese, all 956 1864 1559 76 530 950 973 725 217 M 1393 651 556 18.0 51.0
types
Egg, whole 778 1091 863 301 416 709 515 634 184 M 847 754 301 12.4 74.0
a
Chemical determination.
M
Microbiological determination.
28 Amino Acids
Table 6. Essential amino acid requirements of humans
Amino acid Suggested patterns of requirements [194], Adult requirement, mg kg−1 d−1
g/100 g protein
Infant School-age Adult FAO/WHO NAS/NCR Rosea Hegstedb
child 1973 1980 (144)
His 1.4 – – – – – –
Ile 3.5 3.7 1.8 10 12 10 10
Leu 8.0 5.6 2.5 14 16 11 13
Lys 5.2 7.5 2.2 12 12 9 10
Met + Cys c 2.9 3.4 2.4 13 10 14 13
d
Phe + Tyr 6.3 3.4 2.5 14 16 14 13
Thr 4.4 4.4 1.3 7 8 6 7
Trp 0.85 0.46 0.65 3.5 3 3 3
Val 4.7 4.1 1.8 10 14 14 11
Total 37.3 32.6 15.2 83.5 91 81 80
a
For men.
b
For women.
c
Cys can partly cover the total S-amino acid requirement.
d
Tyr can partly cover the total aromatic amino acid requirement.

total market value. In animal nutrition, amino
acids are used almost exclusively for their nutri-
tive value.
Addition of small amounts of amino acids to
improve the nutritive value of proteins is known
as supplementation. Both supplementation and
the combination of proteins with complemen-
tary amino acids are used to increase the bio-
logic value of proteins. Usually the supply of at
least one of the essential amino acids lies be-
low the requirement. This, the limiting amino
acid, determines what percentage of the protein
(or, more precisely, its amino acids) can be used
to meet the body’s amino acid requirements. In
most cases, methionine is the first limiting ami-
no acid. Sometimes it is lysine; now and then it
is both together.
The contents of essential amino acids found
in several animal and vegetable foodstuffs are
compiled in Table 5. Considerable variations
may be present in the amino acid contents of
a given foodstuff.
The published requirements for the individ-
ual essential amino acids differ. The values (Ta-
ble 6) usually contain safety factors and there-
fore are higher than the minimum requirement.
Requirement values were first determined by
Rose [192]; those published by Hegsted [193]
are considered the most reliable at present. The
amino acid requirement pattern suggested by the
FAO/WHO [194] is considered optimal for the
greatest part of the population.
The “average safe level of daily protein in-
take for men and women,” based on these amino
acid requirement figures, is given as 0.55 g/kg
body weight. The acute daily protein require-
ment, however, varies between 0.5 and 2.5 g/kg
with age and constitution [195]. The Deutsche
Gesellschaft für Ernährung (DGE) recommends
a daily consumption of 0.9 g/kg body weight
[196, 197]. The Committee on Dietary Al-
lowances, Food and Nutrition Board of the Na-
tional Academy of Sciences (NAS), USA, cites
0.8 g/kg as a desirable level of daily protein con-
sumption [198, 199].
5.1.1. Supplementation
In general, animal protein contains the essential
amino acids in larger quantities and in a more
favorable ratio than vegetable protein, which is
often deficient in essential amino acids. Lysine
is the first limiting amino acid in wheat, rye, bar-
ley, oats, maize, and millet, whereas methionine
is the first limiting amino acid in meat, milk,
soybeans, and other beans. The second limiting
amino acids are usually threonine (wheat, rice)
and tryptophan (maize, rice, casein). The limit-
ing amino acids for several foodstuffs are listed
in Table 7.
Improving the biologic value of vegetable
protein in human nutrition is practiced for eco-
nomic and dietary reasons. Combining differ-
ent protein types is not always practical. Often
a complementary protein is unavailable, too ex-
pensive, or not of acceptable taste. In these cases
supplementation with amino acids is the sim-
plest method of increasing the biologic value of
 Amino Acids 29

proteins. There is a monumental volume of lit- tein levels, and the protein efficiency is then
erature on the subject of amino acid supplemen- determined using regressional analysis.
tation [200–207]. The net protein retention (NPR) method and
The biologic value is an important criterion the net protein utilization (NPU) method are
for the evaluation of proteins or amino acid mix- more accurate than the PER method because
tures. It can be determined experimentally [208, they consider the protein (NPR) or total nitro-
209]. In principle, all methods measure the abil- gen (NPU) requirement for maintenance. The
ity of the nutritional protein to replace body pro- “chemical score,” in which the availability of
tein. Table 8 lists the biologic values of nutri- the amino acids is not considered, is suitable
tional proteins as determined by the minimum for a gross estimation of the biologic protein
requirement [195]. Whole egg protein is the ref- quality. In this method the amino acid content
erence in this scale. is determined analytically and compared with
the amino acid pattern of a reference protein,
Table 7. Limiting amino acids in foodstuffs e.g., the FAO/WHO provisional scoring pattern
Proteins First limiting Second limiting
(Table 9). This method provides an immediate
amino acid amino acid(s) picture of the size of amino acid gaps and the
Peanut Thr Lys and Met sequence of limiting amino acids.
Fish Met Lys
Casein Met Trp Content of amino acid in test protein
Score = ×100
Torula yeast Met Content of amino acid in reference protein
Sesame Lys
Skim milk Met
Beans Met Table 9. FAO/WHO provisional scoring pattern 1973 [194]
Sunflower seed Lys Thr Amino acid g/100 g protein
Soy protein Met Lys l-Isoleucine 4.0
Wheat Lys Thr l-Leucine 7.0
Rice Lys Thr and Trp l-Lysine 5.5
Rye Lys Thr and Trp l-Methionine + l-cystine * 3.5
Gelatin Trp l-Phenylalanine + l-tyrosine **
6.0
Maize Lys Trp and Thr l-Threonine 4.0
l-Tryptophan 1.0
Another scale of evaluation is the protein effi- l-Valine 5.0
ciency ratio (PER). This is the daily weight gain Total 36.0
* Cys can partly cover the total S-amino acid requirement.
of young animals, usually rats, under standard ** Tyr can partly cover the total aromatic amino acid requirement.
feeding conditions (see Table 8). In an improved
method, the animals are fed diets of various pro- As can be seen in Table 10, addition of ca.
0.1–0.5 % of the limiting amino acid to such ba-
sic foodstuffs as wheat, rice, maize, and soy-
Table 8. Protein quality of food and minimum requirements (human)
Food Biologic Minimum Protein
value requirement * , efficiency
g kg−1 d−1 ratio (rat)
[195] [195] [190]
Whole egg 100 35 3.92
Beef 92 39 2.30
Cow’s milk 88 40 3.09
Potato 86 [210] 41 [210] ** 3.0 [211]
Fish 3.55
Casein 2.50
Soybean 84 42 2.32
Rice 81 44 2.18
Rye flour 76 46
Maize 72 49 1.18
Beans 72 49 1.48
Wheat flour 56 63 0.6
* Of the protein part.
** Calculated.
30 Amino Acids
Table 10. Increase of the protein efficiency ratio (PER) by supplementation with amino acids [190, 203]
Food (protein content) l-Lys·HCl, l-Thr,
% %
Wheat flour (10 %)
0.2
0.4
0.4 0.15
Rice (7.8 %)
0.2 0.1
Maize (8.75 %)
0.4
Soybean milk (10 %)
Extruded soy protein (10%)
* Reference protein: casein, PER=2.50.
beans raises the protein efficiency in rat growth
tests impressively.
Clinical studies [212–214] and field trials
have solidified the evidence of the benefits to
human nutrition of amino acid supplementation
[215–218]. However, the general supplementa-
tion of basic foodstuffs, such as bread and rice,
is not yet practiced extensively. In dietary nutri-
tion, however, supplementation already plays an
important role (Table 11).
Some infants exhibit lactose or cow’s milk
protein incompatibility. The formulas marketed
for this condition often are based on isolated
soybean protein and are supplemented with l-
methionine to increase the biologic value. The
advantageous effects of l-methionine supple-
mentation on the physical development of in-
fants has been demonstrated in a series of clin-
ical studies [219, 220]. Additionally, the food
for pregnant and nursing women, seniors, over-
weight persons, and athletes can also be supple-
mented. Extruded soy protein, which is used in
large quantities as a meat extender and vegetar-
ian meat substitute, can be supplemented with
N-acetyl-l-methionine [221].
5.1.2. Flavorings, Taste Enhancers, and
Sweeteners
Free amino acids occur in almost all protein-
based foods. In some foods their concentration
is several percent. Foodstuffs having a relatively
high concentration of free amino acids include
fruit juices [222], cheese [223, 224], beer [225],
and seafood [226]. Approximately 85 % of the
free amino acids in orange juice is proline, argi-
d,l-Trp, d,l-Met, PER *
% %
0.65
1.56
1.63
2.67
1.50
2.61
1.41
0.07 2.33
2.12
0.3 3.01
1.99
0.23 2.62
nine, asparagine, γ-aminobutyric acid, aspartic
acid, serine, and alanine [227].
Amino acids are relatively tasteless.
Nonetheless, they contribute to the flavor of
foods. They have characteristic synergistic
flavor-enhancing and flavor-modifying prop-
erties, and they are precursors of natural aro-
mas [228–230]. Amino acids and protein hy-
drolysates are therefore useful additives in the
food industry. The sodium salt of l-glutamic
acid (MSG) exhibits a particularly pronounced
flavor-enhancing effect, leading to the introduc-
tion of a fifth human taste concept of umami
[231], and has been recognized as a flavoring
factor for seaweed, sake, miso, and soy sauce
since 1908. The substance is used in concen-
trations of 0.1–0.4 % as an additive for spices,
soups, sauces, meat, and fish, usually in com-
bination with nucleotides [230]. In 2000, the
existence of a specific receptor for umami was
confirmed [232].
L-Cysteine especially enhances the aroma of
onion [233] and is therefore used to rearomatize
dried onions. Glycine, which has a refreshing,
sweetish flavor, occurs abundantly in mussels
and prawns. It is considered to be an important
flavor component of these products. When used
as an additive for vinegar, pickles, and mayon-
naise, it attenuates the sour taste and lends a note
of sweetness to their aroma. D,L-Alanine is used
for the same purpose in the Far East [234]. Gly-
cine is used to mask the aftertaste of the sweet-
ener saccharin [235, 236].
The l- and d-amino acids usually exhibit pro-
nounced flavor differences. Many l-enantiomers
taste weakly bitter, whereas their optical an-
tipodes, the d-amino acids, taste sweet [237–
 Amino Acids 31
Table 11. Amino acids in dietetic products
Protein/protein hydrolysate Supplemented amino acid Use Indication
Cow’s milk, casein, whey protein l-Cyss and/or l-Lys · HCL infant nutrition adapted nutrition
Soy protein l-Met infant nutrition lactose incompatibility and
milk protein allergy
Casein/yeast l-Lys · HCl meal supplement protein malnutrition, in place
of conventional nutrition

239] (Table 12). For the most part, dipep- pyridines, imidazoles, pyrazines, quinoxalines,
tides and oligopeptides have bitter flavors. One thiophenes, thiolanes, trithianes, thiazoles, and
of the few exceptions is the methyl ester of oxazoles.
the dipeptide l-aspartyl-l-phenylalanine (As- It is often possible to assign certain aro-
partame) [240, 241], which is 150–200 times mas to specific amino acids [252]. For exam-
sweeter than sucrose ( → Sweeteners). ple, the sulfur-containing amino acid cysteine is
primarily responsible for the formation of meat
Table 12. Tastes of l- and d-amino acids* [239] flavor. Proline seems to be important for the
l-Amino acid d-Amino acid aroma of bread crust. Phenylalanine, as well as
Alanine sweet (12–18) sweet (12–18) the branched-chain amino acids leucine and va-
Arginine bitter neutral line, is important for the characteristic flavor of
Asparagine neutral sweet (3–6)
Aspartic acid acidic/neutral acidic/neutral chocolate. Valine and leucine are also involved
Cysteine sulfurous sulfurous in the aroma of roasted nuts. Methionine plays
Glutamine neutral sweet (8–12) a key role in the aroma of French fries. The fla-
Glutamic acid acidic/“glutamate-like”acidic/neutral
Glycine sweet (25–35) sweet (25–35)
vors of such products as precooked foods, snack
Histidine bitter sweet (2–4) articles, and spices may be improved by addi-
Isoleucine bitter sweet (8–12) tion of the proper Maillard aromas. One vari-
Leucine bitter sweet (2–5) ation is adding the precursors of the Maillard
Lysine sweet/bitter sweet
Methionine sulfurous sweet/sulfurous
aromas, i.e., amino acid plus sugar, to the food-
(4–7) stuff and allowing the fragrance to form in situ.
Phenylalanine bitter sweet (1–3) Some aroma profiles that can be prepared from
Proline sweet/bitter (25–40) neutral
amino acids are compiled in Table 13.
Serine sweet (25–35) sweet (30–40)
Threonine sweet (35–45) sweet (40–50)
Tryptophan bitter sweet (0.2–0.4) Table 13. Amino acids for Maillard flavors *
Tyrosine bitter sweet (1–3) Meat, poultry [253] Cys, Cyss, Gly, Glu, Ala, Met,
Valine bitter sweet (10–14) His, Ser, Asp, Pro
* Threshold values for sweet taste in parenthesis, µmol/mL; the Bread, cracker, biscuit [247, Pro, Lys, Arg, Val, His, Leu, Glu,
threshold value for sucrose is 10–12 µmol/mL. 254] Phe, Asp, Gly, Gln
Chocolate, cocoa [255] Leu, Phe, Val, Glu, Ala
The free amino acids are used widely in food- Honey [256–258] Phe
stuff technology as precursors for aromas and Cream, butter [259] Pro, Lys, Ala, Gly, His
Nut, peanut [260] Leu, Val, Ile, Pro, Glu, Gln, His,
brown food colors [242]. The flavors are formed Phe, Asp, Asn
during foodstuff production, e.g., during the Potato [261] Met
ripening of cheese [243, 244], the fermentation Tobacco [257, 262] Asn, Arg, GABA, Gln, Ala, Gly,
Orn, Glu, Asp, Leu, Val, Thr,
of alcoholic beverages [245, 246], or the leav- Pro, Tyr, Phe
ening of dough [247, 248], or foodstuff cook- * Key amino acids underlined.
ing, e.g., frying, roasting, boiling, by the Mail-
lard reaction between amino acids and reduc-
ing sugars (nonenzymatic browning) [228, 249,
250]. The Strecker degradation of amino acids 5.1.3. Other Uses in Foodstuff Technology
plays a central role in this process. A broad spec-
Amino acids are used in the foodstuff industry
trum of aroma-intensive, volatile compounds
for purposes other than supplementation and fla-
forms [251, 252]. The most important classes are
voring. l-Cysteine, for example, is used by the
aliphatic carbonyl compounds and heterocycles,
baked goods and pasta industry as a flour addi-
such as furans, pyrones, pyrroles, pyrrolidines,
tive [263, 264]. As a reducing agent, it relaxes
32 Amino Acids
wheat gluten proteins (by cleavage of the disul-
fide linkages), homogenizes the dough, acceler-
ates dough development, and improves the struc-
ture of the baked product, while allowing shorter
kneading times.
Because they are capable of forming com-
plexes with metals, amino acids act as an-
tioxidants for fats and fat-containing foodstuffs
[265]. This effect is strengthened by primary an-
tioxidants, such asα-tocopherol. Melanoidines,
which are formed during the Maillard reaction,
are stronger antioxidants than the amino acids
themselves [266]. Maillard products are also re-
ported to be preservatives [267]. Glycine appar-
ently exhibits a special preservative effect [268].
There has been a recent upsurge in the sale of
sports drinks and “energy bars”, foodstuffs con-
taining supplements with selected amino acid.
These drinks were originally designed for per-
formance athletes, but are now in widespread
use, and the market is growing rapidly [269].
Amino acids used in functional beverages and
foods include creatine, which is intended to help
build muscle; the branched-chain amino acids
l-isoleucine, l-leucine, and l-valine, which are
important components of cell membrane pro-
teins and assist synthesis of body protein and
reduce central fatigue; and l-arginine, which is
a regulatory agent for the circulation (produc-
tion of NO). These amino acids have a higher
rate of uptake in the body than protein, and im-
prove muscle strength and recovery after exer-
cise [270].
5.2. Animal Nutrition
The use of amino acids for the nutrition of mono-
gastric animals is based on the same foundation
as the supplementation of human foodstuffs and
the clinical experience with humans. In practice,
the enrichment of animal feeds and formulated
feeds with amino acids, especially methionine
and lysine, represents far greater quantities than
does human nutrition. By supplementing feeds
or formulated feeds with the first limiting amino
acid an obvious cost reduction can be achieved,
while maintaining the quality of the ration [271].
Of the ca. 20 amino acids found in feed pro-
tein, about one half are essential for monogastric
animals (see Table 4). Most natural feeds are rel-
atively poor in methionine and lysine (Table 14).
The requirement of our livestock, however, is
comparatively high [272].
When formulating a feed mix for a given an-
imal type, the manufacturer has two choices for
meeting the requirement of a particular amino
acid. He may use either an excess of feed pro-
tein that contains large amounts of this amino
acid or a minimum of natural protein and sup-
plement it with synthetic amino acid. Because
methionine, lysine, and threonine are commer-
cially available and inexpensive, they are often
used in formulated feed. l-Tryptophan, which in
many cases is the third or fourth limiting amino
acid, is becoming more popular as an ingredient
for feed supplements, particularly for pigs.
Amino Acids Content of Feedstuffs. Effec-
tive supplementation requires an exact knowl-
edge of the natural amino acid content of both
the individual feedstuffs and the formulated feed
mix: the desired rates of supplementation must
always be capable for being measured analyt-
ically. Ion-exchange and high-pressure liquid
chromatography are reliable and proven meth-
ods for this.
The amino acid contents of individual feed-
stuffs are published internationally in a large se-
ries of tabulations (see, e.g., Table 14). However,
such data must be current and reliable.
Amino Acid Requirements of Livestock.
Determining amino acid requirement of animals
requires difficult, time-consuming experiments.
The values derived from these experiments are
not constants valid for all times but vary depend-
ing on environmental (heat stress, disease), ge-
netic (sex, breed), and dietary factors (protein
level, energy level, feed intake). There are es-
sentially three methods for determining these re-
quirements: carcass or milk analysis, synthetic
rations, and semisynthetic rations.
For the first method, the amino acid content of
a carcass is taken as a first-order approximation
to the amino acid requirement of an animal. The
same method can be applied for young suckling
mammals by analyzing the amino acid content
of the milk.
In the second method, the test animal is fed a
synthetic mixture of all amino acids, along with
an otherwise balanced ration (control group).
The requirement for a single amino acid is deter-
mined by reducing its content in the diet to zero
 Amino Acids 33
Table 14. Amino acid composition of feedstuffs, wt % [272, 273]
Feedstuff Dry Crude Met Met+Cys Lys Thr Trp Arg Leu Ile Val His
matter protein
Alfalfa 88 17.7 0.27 0.50 0.86 0.76 0.25 0.81 0.72 1.27 0.91 0.36
Barley 88 10.6 0.18 0.42 0.38 0.36 0.12 0.53 0.37 0.73 0.52 0.24
Beans, field 88 25.4 0.20 0.52 1.63 0.90 0.22 2.29 1.03 1.89 1.15 0.67
Blood meal 91 88.8 1.03 2.17 7.96 3.85 1.42 3.94 1.17 11.39 7.69 5.65
Corn 88 8.5 0.18 0.37 0.25 0.31 0.06 0.40 0.29 1.05 0.41 0.26
Corn gluten 88 19.0 0.32 0.71 0.58 0.68 0.11 0.85 0.59 1.69 0.89 0.59
feed
Corn gluten 88 60.6 1.43 2.52 1.02 2.08 0.31 1.93 2.48 10.19 2.79 1.28
meal
Feather meal 91 81.1 0.61 4.74 2.08 3.82 0.54 5.62 3.86 6.79 5.88 0.93
Fish meal 91 62.9 1.77 2.34 4.81 2.64 0.66 3.66 2.57 4.54 3.03 1.78
Meat and bone 91 49.1 0.68 1.18 2.51 1.59 0.28 3.45 1.34 2.98 2.04 0.91
meal
Meat meal 91 48.8 0.68 1.24 2.44 1.63 0.30 3.42 1.40 2.99 2.13 0.93
Oat 88 12.6 0.22 0.57 0.53 0.44 0.14 0.87 0.48 0.92 0.66 0.31
Rapeseed 88 34.8 0.70 1.59 1.95 1.53 0.45 2.15 1.37 2.47 1.77 0.97
meal,
Rice 88 7.3 0.20 0.37 0.26 0.26 0.09 0.60 0.29 0.59 0.40 0.18
Sesame meal 88 41.1 1.15 1.97 1.01 1.44 0.54 4.86 1.47 2.74 1.85 0.98
Sorghum 88 9.3 0.17 0.34 0.22 0.31 0.10 0.38 0.37 1.21 0.46 0.23
Soybean meal, 88 44.0 0.64 1.31 2.75 1.76 0.57 3.28 2.01 3.44 2.09 1.21
44 % CP*
Soybean meal, 88 47.6 0.69 1.41 2.98 1.89 0.61 3.52 2.16 3.71 2.23 1.31
48 % CP
Sunflower 88 33.5 0.77 1.36 1.19 1.25 0.40 2.75 1.37 2.15 1.66 0.88
meal
Tapioca 88 3.3 0.04 0.09 0.12 0.11 0.04 0.18 0.11 0.19 0.14 0.08
Triticale 88 11.6 0.21 0.49 0.42 0.39 0.12 0.61 0.42 0.79 0.55 0.29
Wheat 88 13.3 0.21 0.50 0.38 0.38 0.15 0.64 0.44 0.87 0.56 0.32
Wheat bran 88 15.7 0.25 0.58 0.64 0.52 0.22 1.07 0.49 0.98 0.72 0.44
Wheat gluten 88 14.4 0.22 0.52 0.46 0.46 0.19 0.86 0.45 0.89 0.67 0.39
feed
Wheat gluten 88 74.3 1.17 2.79 1.24 1.89 0.68 2.59 2.65 5.20 2.88 1.54
meal
* Crude Protein

and from there on supplementing it stepwise up
to an amount where the animal performs as well
as the control group.
The third and most commonly used method
is that a basal diet consisting of typically used
feeds is formulated to be deficient in one ami-
no acid but adequate in all other nutrients. To
this basal diet graded levels of the amino acid in
question are added until the performance of the
animal approaches a maximum. The response
in growth to stepwise increasing of amino acid
supplementation follows the law of diminishing
returns and can be described best by an expo-
nential function.
Amino acid recommendations and require-
ments vary between species and for each species
with age. There are several physiological rea-
sons for that:
The various proteins deposited in the ani-
mal’s body differ considerably in their amino
acid composition. In poultry, for example, the
proportion of lysine is much higher in mus-
cle protein than in feather protein. In contrast,
methionine and cystine are required in a higher
percentage for the formation of feather protein
and for maintaining metabolic functions than for
muscle protein.
Not only the amino acid composition of the
different proteins varies but also the quantity of
each protein deposited in the animal changes
with age. The daily accretion of muscle protein
increases up to a certain age depending on the
species and declines thereafter. The daily pro-
tein requirement for metabolic functions rises
continually as the animal grows. Another rea-
son for amino acid recommendations changing
with age is the fact that young animals have a
high potential for growth but at the same time
a relatively small feed intake capacity which re-
quires a high nutrient density in the diet.
34 Amino Acids
Table 15. Amino acid recommendations, wt %, for poultry [273] and pigs [274, 275]
Species Metabolizable Crude Met Met+ Lys Thr Trp
energy, MJ/kg protein Cys
Broiler
Starter 12.7 21 0.57 0.92 1.27 0.80 0.20
Grower 13.2 18 0.44 0.76 1.00 0.65 0.19
Finisher 13.2 17 0.41 0.74 0.95 0.63 0.11
Laying hen (105 g feed intake per 12.0 16 0.38 0.71 0.80 0.52 0.15
day)
Turkey (weeks of age)
0–4 11.7 28 0.66 1.15 1.80 1.06 0.31
5–8 12.1 25 0.59 1.04 1.60 0.95 0.27
9–12 12.6 22 0.53 0.92 1.40 0.84 0.24
13–16 13.0 19 0.46 0.80 1.20 0.73 0.20
> 16 13.4 16 0.43 0.75 1.10 0.67 0.19
Pig (kg live weight)
10–19 10.4 18 0.48 0.87 1.40 0.94 0.25
20–30 10.2 17 0.40 0.74 1.15 0.79 0.23
40–70 9.8 14.5 0.35 0.64 0.95 0.68 0.19
70–105 9.6 13.5 0.30 0.55 0.82 0.59 0.16
Sow
Lactating 9.8 16.5 0.36 0.65 1.00 0.70 0.20
Pregnant 8.9 12.5 0.23 0.42 0.70 0.46 0.14

The amino acid recommendations for domes-
tic monogastric animals are listed in Table 15.
Economics of Amino Acid Supplementa-
tion. The purpose of modern, formulated feed
mixes is to meet all nutritional requirements of
the animal at a minimum cost, and amino acids
and proteins are usually among the most expen-
sive components of a feed mix. The performance
of a feed mix is measured primarily by feed uti-
lization and weight gain. Both feed utilization
and weight gain, as well as other factors such as
laying performance or feather or hair growth, are
directly dependent on a sufficient supply of ami-
no acids. Methionine and lysine play the major
roles. Methionine in the form of d,l-methionine
and lysine in the form of l-lysine · HCl are used
in place of or as a supplement to natural methi-
onine and lysine sources [276].
Linear programming is the method of choice
for simultaneously optimizing a ration and min-
imizing cost [277]. This method allows simul-
taneous consideration of all demands that are
made on the ration. A prerequisite is exact data
on the nutrient content of all available feedstuffs
and additives as well as their prices and avail-
abilities. Additionally, the restrictions, i.e., the
dietary requirements that the ration has to fulfill,
must be known. Table 16 shows two examples
of feed mixes formulated by the linear program-
ming method commonly used.
Protein and Amino Acid Digestibilities of
Feed Ingredients. The nutritive value of pro-
tein for monogastric animals is determined not
only by the amino acid composition of the diet
but also by digestibility of the individual amino
acids, in particular the amino acids likely to be
limiting. Over the last decades, considerable re-
search has been carried out that demonstrates
large differences in amino acid digestibilities
between feeds.
Amino acid digestibilities can be determined
according to the ileal or fecal analysis method.
The fecal analysis method measures the amount
of each amino acid consumed and excreted in
feces. The amino acid digestibilities determined
according to the ileal analysis method are cal-
culated based on the intake and amount of each
amino acid passing at the end of the distal ileum.
The ileal analysis method should be considered
as an improvement on the fecal analysis method,
since the protein or amino acid absorbed in the
large intestine make little or no contribution to
the protein status of the animal. The ileal anal-
ysis method is also very sensitive to differences
in amino acid digestibilities, as these result from
processing conditions or from inherent differ-
ences between samples of the same feedstuff.
Digestibilities measured over the entire diges-
tive tract may be altered by resident bacteria.
The bacteria may break down some of the ami-
no acids, convert them to other amino acids, or
even produce new amino acids. Measuring ami-

Table 16. Two examples of formulated feeds
Broiler feed composition, wt %
Yellow corn 28.00
Wheat 20.00
Soybean meal (48 % crude protein) 30.00
Tapioca meal 10.00
Fat 7.00
Meat and bone meal (45 % crude 3.00
protein)
Mineral premix 1.25
Vitamin-trace element premix 0.50
d,l-Methionine 0.25
100.00
no acids excreted in the feces will not reflect
unabsorbed amino acid, but rather unabsorbed
amino acids after possible alteration by the bac-
teria.
Amino acid digestibility values from the liter-
ature, determined with the ileal analysis method
(Table 17) show large differences in amino acid
digestibilities between feeds and among differ-
ent samples of the same feed.
The differences in amino acid digestibility of
feeds can be attributed to various factors. These
include, e.g., fiber levels, heat damage during
processing and the presence of antinutritional
factors that interfere with nutrient digestion and
utilization.
Studies have clearly shown improvements in
diet formulation practices, when diets are for-
mulated on the basis of ileal digestible rather
than on supply of limiting amino acids. This
holds especially true when a good quality pro-
tein supplement is replaced by protein supple-
ment(s) of lower quality combined with sup-
plementary amino acids. In consequence, amino
acid requirements should be expressed as ileal
digestible, rather than as total amino acid re-
quirements.
Amino Acids as a Measure to Reduce the
N-Output from Livestock Production. Ani-
mal production accounts for a significant portion
of nitrogen containing compounds released into
the environment. In areas with intensive live-
stock production this might result in environ-
mental problems, especially if N-requirements
for crop fertilization and N-output from live-
stock production get out of balance. In this case
nitrogen containing compounds are released into
Amino Acids 35
Pig fattening feed composition, wt %
Feed grain (barley, wheat, corn) 35.00
Soybean meal (44 % crude protein) 19.00
Tapioca meal 20.00
Corn gluten feed 15.00
Meat and bone meal (45 % crude 3.00
protein)
Fat 3.00
Beet molasses 2.00
Mineral premix 2.43
Vitamin-trace element premix 0.50
d,l-Methionine 0.07
100.00
the surface and ground water where they accu-
mulate.
The total amount of nitrogen produced is de-
pendent on both the average number of animals
per unit available on and the efficiency of con-
version of feed protein into body protein. This
efficiency is often impaired due to diets that are
not balanced for the specific amino acid require-
ments of the animal fed. The portion of protein
that is fed without supplying an adequate mix
of amino acids is thus excreted without being
utilized by the animals. Meanwhile the animal’s
requirements for amino acids are well known
which allows a reduction in the total protein
content of diets as long as the diet is supple-
mented with amino acids in accordance with the
animal’s specific requirements. As a result, feed
protein utilization is maximized and also water
intake as a means to excrete excess nitrogen via
the kidney is reduced.
In consequence the N-output from livestock
production can be reduced down to 65 % when
supplemental amino acids are used together with
a reduction of the protein content of the diet.
5.3. Pharmaceuticals
The pharmaceutical industry requires amino
acids at a rate between 2000 and 3000 t/a world-
wide. More than half of this is used for infusion
solutions. During the last few years the poten-
tial of amino acids and their derivatives as active
ingredients for pharmaceuticals has been recog-
nized clearly, and considerable growth can be
predicted.
36 Amino Acids
Table 17. Range of ileal digestibilities (%) of lysine, methionine, and threonine in different feeds for pigs modified from [274]
Amino acids Lysine
Cereal grains
Barley 65–79
Wheat 62–81
Corn 71–82
Protein supplements
Soybean meal (44 % CP* ) 85–89
Canola meal 74–76
Meat and bone meal 58–67
Cottonseed meal 53–70
* Crude Protein
5.3.1. Nutritive Agents
Infusion Solutions. Parenteral nutrition
with l-amino acid infusion solutions is a well-
established component of clinical nutrition
therapy. A standard infusion solution contains
the eight classical essential amino acids, the
semiessential amino acids l-arginine and l-
histidine, and several nonessential amino acids,
generally glycine, l-alanine, l-proline, l-serine,
and l-glutamic acid.
Also available are special infusion solutions
tailored to the requirements of particular groups,
such as newborn infants, seniors, or patients
with an extreme negative nitrogen balance. So-
lutions rich in the branched-chained amino acids
leucine, isoleucine, and valine and poor in methi-
onine and aromatic amino acids are available for
liver-disease patients. Solutions containing only
essential amino acids are available for kidney
patients. Enzymatic protein hydrolysates, which
were used as infusion solutions until the 1980s
have disappeared almost completely from the
market. They were not available in the optimal
composition, and there were often compatibility
problems. Only pure, crystalline l-amino acids
are used in modern infusion solutions. The so-
lutions (up to 10 %), which also contain elec-
trolytes in addition to amino acids, are sterile
and pyrogen-free.
The simultaneous administration of carbo-
hydrates is necessary for optimal utilization of
the amino acids. Glucose is normally a separate
infusion. Some commercially available amino
acid infusion solutions contain an energy source
in the form of sugar alcohols (sorbitol, xylitol),
which do not enter into a Maillard reaction with
the amino acids.
Normally, parenteral nutrition is only prac-
ticed over a limited time. In principle, however,
Methionine Threonine
72–88 64–76
79–92 51–78
88–92 74–79
77–90 73–81
81–87 66–67
72–79 53–62
65–82 55–69
total parenteral nutrition over many years is pos-
sible. In such a case, all essential nutrients (un-
saturated fatty acids, vitamins, and trace ele-
ments) must be provided.
Elemental Diets. Enteral nutrition is also a
means of providing the essential nutrients [278].
Elemental diets, which were developed origi-
nally for the astronauts [389], contain chemi-
cally defined nutritive components. In addition
to free amino acids the mixtures generally con-
tain carbohydrates, fats, minerals, and vitamins
in a combination adapted to the requirements.
In many cases, elemental diets are used as an al-
ternative and supplement to parenteral nutrition.
They have high nutritional value and are totally
resorbable. They are largely independent of the
digestive function of the pancreas and reduce the
intestinal bacteria flora. Amino acid elemental
diets generally are used in cases of anatomic,
functional, or enzymatic defects [279].
Formula diets based on peptides are gaining
ground as an alternative to elemental diets based
on l-amino acids. According to [280], short-
chained peptides are resorbed rapidly via a pep-
tide transport system in the gut, therefore in a
process that is independent of amino acid trans-
port. Compositions of nitrogen-free amino acid
analogues (keto acids and hydroxy acids) have
come into use for the special case of kidney in-
sufficiency (chronic renal failure).
Elemental diets or formula diets are admin-
istered orally or via a nasogastric tube directly
into the gastrointestinal tract.
5.3.2. Therapeutic Agents
Many therapeutic agents are derivatives of
natural or nonnatural amino acids. Exam-
ples are benserazide, captopril, and dextrothy-
roxine. They are described under keywords

such as → Spasmolytics, → Antihypertensives,
→ Antihypotensives, or → Thyrotherapeutic
Agents. Only therapeutically useful amino acids
and simple derivatives are treated here.
Amino Acids and Salts. The amino acids
and their simple salts that are important thera-
peutic agents are compiled in Table 18. The pro-
prietary names listed represent only a selection.
N-Acetylcysteine [616-91-1], C5 H9 NO3 S,
20
M r 163.2, mp 109–110 ◦ C, [α]D + 5◦ (c = 3,
H2 O), is a mucolytic and secretolytic agent. It is
also indicated for the treatment of paracetamol
poisoning.
N-Acetylcysteine is prepared by reaction of
cysteine hydrochloride monohydrate with acetic
anhydride in the presence of sodium acetate
[281, 282].
Trade names: Exomouc (Bouchara),
Fluimucil (Zambon) and numerous others [283].
Carbocisteine (carbocysteine) [638-23-3],
S-carboxymethyl-l-cysteine, C5 H9 NO4 S, M r
20
179.2, mp 204–207 ◦ C (decomp.), [α]D −34.0
◦
to −36.0 (c = 10, H2 O), is used to treat disor-
ders of the respiratory tract associated with ex-
cessive mucus.
Synthesis involves S-alkylation of l-cysteine
with chloroacetic acid in the presence of sodium
hydroxide [284, 285].
Trade names: Mucodyne (Sanofi-Aventis)
and numerous others [283].
Fudosteine [13189-99-5], S-(3-hydroxypro-
pyl)- l-cysteine, C6 H13 NO3 S, M r 179.24, mp
198–202 ◦ C, [α]20
D −22
◦
(c = 1, H2 O), was
launched by Mitsubishi Pharma in 2001 for the
treatment of bronchial hypersecretion.
Amino Acids 37
Fudosteine is synthesized by condensation of
l-cysteine with 3-bromopropyl alcohol in aque-
ous NaOH, or by condensation of l-cysteine
with allyl alcohol by means of aqueous potas-
sium persulfate [286].
Trade names: Cleanal, Spelear (Mitsubishi
Pharma).
Levodopa [59-92-7], (S)−3-(3,4-dihy-
droxyphenyl)alanine, C9 H11 NO4 , M r 197, mp
20
285.5 ◦ C (decomp.), [α]D −12.15◦ (c = 4 in
1 M HCl).
Levodopa is widely used for treatment of
Parkinson’s disease, most often in combination
with peripheral decarboxylase inhibitors such as
benserazide and carbidopa. For manufacture and
trade names → Parkinsonism Treatment.
Trade names: Larodopa (Roche) and numer-
ous others [283].
Mecysteine hydrochloride [18598-63-5],
cysteine methyl ester hydrochloride, methyl l-
2-amino-3-mercaptopropionate hydrochloride,
C4 H10 ClNO2 S, M r 171.66, mp 140–141 ◦ C, is
used in the treatment of disorders of the respi-
ratory tract associated with excessive mucus. It
is prepared by esterification of l-cysteine hy-
drochloride monohydrate with methanol in the
presence of hydrogen chloride [287].
Trade names: Acthiol (Joullié, France), Ac-
tiol (Lirca, Italy), Visclair (Sinclair, UK), Aslos-
C (Nissin, J), Epectan (Seiko, Japan), Moltanine
(Tohok.-Tokyo Tanabe, Japan), Radcol (Nip-
pon Universal, Japan), Sekinin (Tokyo Hosei,
Japan).
Methiosulfonium chloride (iodide) [1115-
84-0], L-methylmethionine sulfonium chlo-
ride, vitamin U, C6 H14 ClNO2 S, M r 199.7;
mp 134 ◦ C (decomp.). Iodide [3493-11-6],
C6 H14 INO2 S, M r 291.1.
38 Amino Acids
Table 18. Amino acids and their salts as drugs
Compound Formula Mr Medical use Trade names
l-Alanine [56-41-7] C3 H7 NO2 89.09 parenteral nutrition, stimulant of glucagon
secretion
l-Arginine [74-79-3] C6 H14 N4 O2 174.20 parenteral nutrition, stimulation of pituitary
release of growth hormone and prolactin,
stimulation of pancreatic release of glucagon
and insulin
l-aspartate [7675-83-4] C10 H21 N5 O6 307.31 treatment of hyperammonemia Argihepar, Laevil,
Potentiator, Sargenor,
Sorbenor
l-glutamate [4320-30-3] C11 H23 N5 O6 321.34 treatment of hyperammonemia Modumate
hydrochloride [1119-34-2] C6 H15 ClN4 O2 210.66 treatment of hyperammonemia, electrolyte Argivene, R-Gene,
concentrate for i.v. solutions Polilevo
l-pyroglutamate C11 H21 N5 O5 303.27
2-oxoglutarate [16856-18-1] C11 H20 N4 O7 320.3 treatment of hepatic disorders Leberam, Anetil,
Argiceto, Eucol
l-Asparagine [70-47-3] C4 H8 N2 O3 132.13 parenteral nutrition
monohydrate [5794-13-8] C4 H10 N2 O4 150.13
d,l-Aspartic acid [617-45-8] C4 H7 NO4 133.10
magnesium, [52101-01-6] C8 H20 N2 O12 Mg 360.54 cardiac agent, management of fatigue, mineral Magnesium Verla,
tetrahydrate supplement
potassium, [923-09-1] C4 H6 NO4 K · 1/2 H21O80.20 cardiac agent, management of fatigue, mineral Trommcardin,
semihydrate (anhydrous) supplement Trophicard-Köhler
sodium, C4 H8 NO5 Na 173.10
monohydrate
l-Aspartic Acid [56-84-8] C4 H7 NO4 133.10 parenteral nutrition
ferrous, C8 H2 ON2 O12 Fe 392.1 treatment of iron deficiency Sideryl, Spartocine
tetrahydrate
magnesium, [2068-80-6] C8 H16 N2 O10 Mg 324.52 management of fatigue and heart conditions
dihydrate
potassium, [1115-63-5] C4 H6 NO4 K · 1/2 H21O80.20 management of fatigue and heart conditions Magnesiocard,
semihydrate
(anhydrous) Corroverlan
sodium, [3792-50-5] C4 H8 NO5 Na 173.10 parenteral nutrition
monohydrate
(anhydrous)
Betaine [107-43-7] C5 H11 NO2 117.15
citrate [17671-50-0] C11 H19 NO9 309.27 lipotropic Flacar, Panstabil
hydrochloride [590-46-5] C5 H12 ClNO2 153.61 lipotropic, gastric acidifier Aciventral, Acidol,
Pesimuriat
monohydrate [17146-86-0] C5 H13 NO3 135.15 lipotropic Hepaderichol
l-Citrulline [372-75-8] C6 H13 N3 O3 175.19 treatment of hepatic disorders Polilevo
l-Cysteine [52-90-4] C3 H7 NO2 S 121.16 treatment of damaged skin, topically in Reducdyn,
ophthalmology, Hepa-Loges,
detoxicant Irradian, Cicatrex,
Felacomp
hydrochloride [52-89-1] C3 H8 ClNO2 S 157.62 Cheihepar, Choldestal
hydrochloride [7048-04-6] C3 H10 ClNO3 S 175.64
monohydrate
l-Cystine [56-89-3] C6 H12 N2 O4 S2 240.30 parenteral nutrition, lipotropic agent, treatment Cystin “Brunner”,
of hair and nail damage Gerontamin,
Pantovipar, Priorin
l-Glutamic acid [56-86-0] C5 H9 NO4 147.13 parenteral nutrition, dietary supplement, Aciglut,
treatment of hyperammonemia Glutamin-Verla
calcium, [5996-22-5] C10 H20 N2 O10 Ca 368.2 Vivacalcium
dihydrate
hydrochloride [138-15-8] C5 H10 ClNO4 183.54 gastric acidifier Bioprotein-Holzinger,
Pansan, Pepsalara
monosodium, [142-47-2] C5 H10 NO5 Na 187.13 flavoring and seasoning of food
monohydrate
(anhydrous)
magnesium, [19238-50-7] C10 H24 N2 O12 Mg 388.62 tonics, mineral supplements Magnesium Verla,
tetrahydrate Glutergen
potassium, [19473-49-5] C5 H10 NO5 K 203.24 tonics, mineral supplements
monohydrate
(anhydrous)
 Amino Acids 39
Table 18. (Continued)
Compound Formula Mr Medical use Trade names
magnesium, C10 H17 BrN2 O8 Mg 397.5 tranquilizer Psicosoma,
hydrobromide Psychoverlan
l-Glutamine [56-85-9] C5 H10 N2 O3 146.15 parenteral nutrition, treatment of mental
disorders and alcoholism
Glycine [56-40-6] C2 H5 NO2 75.07 parenteral nutrition, antacid in conjunction with Cicatrex,
calcium carbonate Felacomp,
Gastripan-K,
Mirogastrin
aluminum, hydrate [13682-92-3] C2 H6 NO4 Al 135.1 antacid Acidrine,
( + x H2 O) Al-Glycin
l-Histidine [71-00-1] C6 H 9 N 3 O 2 155.16 parenteral nutrition, essential amino acid for
infants
acetate C8 H13 N3 O4 215.21
hydrochloride [5934-29-2] C6 H12 ClN3 O3 209.63 Anti-rheuma,
monohydrate
Rollkur-
Ankermann,
Laristine,
Plexamine
l-Isoleucine [73-32-5] C6 H13 NO2 131.18 parenteral nutrition, treatment of hepatic coma,
dietary supplement
l-Leucine [61-90-5] C6 H13 NO2 131.18 parenteral nutrition, treatment of hepatic coma, numerous
dietary supplement combinations
d,l-Lysine [70-54-2] C6 H14 N2 O2 146.19 formation of salts with acidic drugs (to enhance
solubility)
monohydrochloride [70-53-1] C6 H15 ClN2 O2 182.65 geriatric Jestrosemin
acetylsalicylate [34220-70-7] C15 H22 N2 O6 326.34 soluble form of acetylsalicylic acid (aspirin) for Aspisol, Delgesic
injection
l-Lysine [56-87-1] C6 H14 N2 O2 146.19 parenteral nutrition, dietary supplement, numerous
prophylaxis of combinations
acetate [57282-49-2] C8 H18 N2 O4 206.24 herpes simplex infection (?)
l-aspartate [27348-32-9] C10 H11 N3 O6 279.30 dietary supplement
l-glutamate [5408-52-6] C11 H23 N3 O6 293.32 dietary supplement
l-malate [71555-10-7] C10 H20 N2 O7 280.28 dietary supplement
monohydrate [39665-12-8] C6 H16 N2 O3 164.21 dietary supplement
monohydrochloride [657-27-2] C6 H15 ClN2 O2 182.65 treatment of hypochloremia, alkaloses Aktivanad,
Athensa,
Omnival, Vivioptal
d,l-Methionine [59-51-8] C5 H11 NO2 S 149.21 lipotropic and choleretic agent numerous
combinations
l-Methionine [63-68-3] C5 H11 NO2 S 149.21 parenteral nutrition, dietary supplement,
lipotropic agent,
treatment of paracetamol poisoning
l-Ornithine
acetate [60259-81-6] C7 H16 N2 O4 192.22 parenteral nutrition
l-aspartate [3230-94-2] C9 H19 N3 O6 265.27 treatment of hepatic disorders Hepa-Merz
monohydrochloride [3184-13-2] C5 H13 ClN2 O2 168.62 parenteral nutrition Ornitaine, Polilevo
2-oxoglutarate [5191-97-9] C10 H18 N2 O7 278.14 treatment of hepatic disorders Ornicetil
(hyperammonemia)
d-Phenylalanine [673-06-3] C9 H11 NO2 165.19 antidepressant
l-Phenylalanine [63-91-2] C9 H11 NO2 165.19 parenteral nutrition
l-Proline [147-85-3] C5 H9 NO2 115.13 parenteral nutrition, dietary supplement,
starting material
for captopril and enalapril
l-Pyroglutamic acid [98-79-3] C5 H7 NO3 129.07 formation of salts with basic drugs
d,l-Serine [302-84-1] C3 H7 NO3 105.09 starting material for benserazide Aktiferrin
l-Serine [56-45-1] C3 H7 NO3 105.09 parenteral nutrition, dietary supplement Sulfolitruw
l-Threonine [72-19-5] C4 H9 NO3 119.12 parenteral nutrition, dietary supplement
l-Tryptophan [73-22-3] C11 H12 N2 O2 204.23 parenteral nutrition, antidepressant, sleep Optimax, Kalma,
inducer, dietary supplement Pacitron
l-Tyrosine [60-18-4] C9 H11 NO3 181.19 parenteral nutrition, dietary supplement
l-Valine [72-18-4] C5 H11 NO2 117.15 parenteral nutrition, dietary supplement,
treatment of hepatic coma
40 Amino Acids
Methiosulfonium chloride is used for its protec-
tive effect on the liver and gastrointestinal mu-
cosa, whereas the iodide finds use for rheumatic
disorders. The compounds are made by heating
l-methionine with methyl chloride or methyl io-
dide [288].
Trade names: Chloride: Ardesyl (Beytout,
France), withdrawn from the market; Cabagin
(Kowa, Japan). Iodide: Lobarthrose (Opodex,
France), withdrawn from the market.
Oxitriptan [4350-09-8], (S)-5-hydroxytryp-
tophan, C11 H12 N2 O3 , M r 220, mp 273 ◦ C (de-
22 22
comp.), [α]D −32.5◦ (c = 1 in H2 O), [α]D +
◦
16.0 (c = 1 in 4 M HCl).
This intermediate in mammalian biosynthesis
of serotonin is used as an antidepressant. It is
produced either by total synthesis (analogous
to l-tryptophan via the 5-benzyloxy derivative)
[289–291] or by fermentation with Chromobac-
terium violaceum [292].
Trade names: Levothym (Promonta Lund-
beck, Germany), Lévotonine (Panpharma,
France), Oxyfan (Coli, Italy), Tript-OH (Sigma-
Tai, Italy).
D-Penicillamine [52-67-5], D-3-mercapto-
valine, d-β,β-dimethylcysteine, C5 H11 NO2 S,
M r 149.21, mp 198.5 ◦ C, [α]D −63◦
25
(c = 0.1, pyridine). Hydrochloride [2219-30-9],
C5 H12 CINO2 S, M r 185.7, mp 177.5 ◦ C (de-
25
comp.) [α]D −63◦ (c = 1 in 1 M NaOH).
d-Penicillamine is a chelating agent that aids
the elimination of toxic metal ions, e.g., cop-
per in Wilson’s disease. It is used, as an alter-
native to gold preparations, in the treatment of
severe rheumatoid arthritis. It is useful in treat-
ing cystinuria because it reacts with cystine to
form cysteine-penicillamine disulfide, which is
much more soluble than cystine.
d-Penicillamine is produced by hydrolysis
of benzylpenicillin (→ Antibiotics, Chap. 4)
via its Hg(II) complex [293] or by total syn-
thesis. In the synthesis, isobutyraldehyde, sul-
fur, and ammonia are condensed to 5,5-di-
methyl-2-isopropyl-∆3 -thiazoline, which, on
reaction with hydrogen cyanide, gives 5,5-di-
methyl-2-isopropyl-thiazolidine-4-carbonitrile.
Hydrolysis with boiling hydrochloric acid yields
d,l-penicillamine hydrochloride. Cyclization
with acetone and formylation leads to d,l-3-
formyl-2,2,5,5-tetramethylthiazolidine-4-car-
boxylic acid, which can be resolved with (−)-
phenylpropanolamine via the diastereomeric
salts. Hydrolysis with hydrochloric acid leads
to d-penicillamine hydrochloride [294].
Trade names: Cuprimine (Merck Sharp &
Dohme, USA), Depen (Carter-Wallace, USA),
Distamine (Dista, UK), Metalcaptase (Heyl,
Germany), Trolovol (ASTA Medica AWD, Ger-
many), Trisorcin (Merdele, Germany), Trolovol
(Bayer, France), Pendramine (ASTA Medica,
UK), Pemine (Lilly, Italy).
5.4. Cosmetics
Amino acids are a major component of the “nat-
ural moisturizing factor” (NMF) that protects
the skin surface from dryness, brittleness, and
a deleterious environment [295]. The epidermis
of the skin contains about 15 % water, which, in
the presence of amino acids, principally serine,
citrulline, and alanine, forms a stable water-in-
oil emulsion with the skin lipids in the form of a
thin layer. The amino acids simultaneously sta-
bilize the pH of the skin (the acidic layer), and
can be absorbed into the skin and hair. Because
of these properties, amino acids [296, 297], pro-
tein hydrolysates [298], and proteins [296] are
widely utilized in skin and hair cosmetics, e.g.,
in mild skin creams, skin cleansing lotions, and
hair shampoos. It has been claimed that 60 %
of hair strength is accomplished by three ami-
no acids, i.e., histidine, lysine, and tyrosine, and
supplementing these in hair shampoos can re-
duce loss of hair strength and improve shine
[299]. Methionine is absorbed into the scalp and
absorbed by the hair fibers as cysteine, which

is responsible for the cross-linking of keratin
[300]. A report published in 2004 claims that
amino acids comprise the fastest growing area of
new ingredients in the moisturizers and humec-
tants market [301]. Amino acids manufactured
by fermentation rather than extracted from an-
imal protein have been increasingly promoted
as being more “natural”, and this has helped to
increase the market for these products.
The sodium salts of the reaction products
of fatty acids with amino acids, such as glu-
tamic acid [302–305], or short-chain peptides
(protein hydrolysates) [306, 307] are surfactants.
They are effective, skin-compatible cleaners and
emulsifiers, which are used in shampoos, shower
gels, baby baths, medicinal skin cleansers, cold-
wave preparations, etc. [303, 308, 309]. Arg-
inine has been used as the cation in soaps to
improve the foam volume, and the free ami-
no acid has been used in conditioners to im-
prove sensory properties [310]. The sulfur-con-
taining amino acids exhibit a special normaliz-
ing effect on skin metabolism, e.g., in cases of
excess skin lipid production (seborrhea), dan-
druff, or acne. Substances utilized for this pur-
pose include derivatives of cysteine (e.g., S-car-
boxymethylcysteine), homocysteine (2-amino-
4-mercaptobutyric acid), and methionine [311].
Amino acids are also used in hair lotions, where
they are reported to have a nutritive effect [312].
Cysteine, which acts as a reducing agent, is gain-
ing importance, especially in Japan, as a sub-
stitute for thioglycolic acid in permanent wave
preparations that are less damaging to hair [313].
Use of aluminum, tin, and zirconium complexes
of amino acids, especially glycine, as deodorants
[314] and antiperspirants [314, 315] has been re-
ported.
The use of peptides as mimics of human
protein fragments in cosmetics has been devel-
oped since 2000. One such product is Matrixyl
(Sederma), a pentapeptide (Palmitoyl-Lys-Thr-
Thr-Lys-Ser), which is a mimic of a precolla-
gen fragment [316]. Matrixyl is applied in anti-
wrinkle creams. Another example is the dipep-
tide N-acetyl-Tyr-Arg-cetyl ester, registered as
Calmosensine (Sederma), which stimulates en-
dorphin precursor release in skin cells and acts
as a mild analgesic [317].
Amino Acids 41
5.5. Agrochemicals
An increasing number of pesticides [318] are
derivatives of natural or nonnatural amino acids.
Important amino acids like Glyphosate or Glu-
fosinate are major products in the agrochemical
market. The synthesis of the active ingredient
may start with an amino acid, but very often the
amino acid moiety is formed during the chemi-
cal synthesis.
5.5.1. Herbicides (→ Weed Control)
All pesticide segments (i.e., herbicides, fungi-
cides, insecticides) contain compounds with
amino acid substructures, but the major applica-
tion of pesticidal amino acids is in the herbicide
area [319].
Glyphosate [1071-83-6] [320], Roundup,
C3 H8 NO5 P, M r 169.1, mp 200 ◦ C, is the iso-
propylamine salt of a phosphorus containing
glycine derivative with a hybrid structure [321].
Its mode of action is unique; it inhibits dif-
ferent enzyme systems that leads to the block-
ing of different amino acid biosyntheses. The
substance is used as a nonselective contact her-
bicide to control deep rooted weeds. The sales
[322] in 1997 grew to about $2180 × 106 and
reaching this figure it is the worlds top selling
pesticide. The growth rates were double digits
in the 1990s (17 %) and for 2002 an annual pro-
duction of 180 000 t was estimated. Additional
driving impact on the sales of glyphosate has
been achieved by the introduction of glyphosate
resistant crops (1996: soy beans, 1997: canola
and cotton, 1998: maize) and the consequent
distribution of genetically engineered seeds in
combination with this herbicide.
The classical synthesis starts with iminodi-
acetic acid, phosphorous trichloride and formal-
dehyde, but also sequences using glycine and
dimethyl phosphate have been applied. There
are no confirmed reports of resistance. The ex-
piration of the original patent and the advent of
generic producers guarantee an increase of pro-
duction of the substance becoming a bulk chem-
ical.
42 Amino Acids

Sulfosate [81591-81-3] [323], Touchdown, to an imidazolinone with an ortho-carboxy sub-

C6 H16 NO5 PS, M r 245.2, is the trimeth-
ylsulfonium salt of Glyphosate also applied as
a nonselective herbicide that was introduced in
1989. As in the case of Glyphosate, sales are
growing well.
Glufosinate [324], Basta, Phosphinothricine
[53369-07-6], C5 H12 NO4 P, M r 181.1, mp
215 ◦ C (for the ammonium salt). This phos-
phorous analogue of glutamic acid was intro-
duced in 1984 as a nonselective herbicide in
the speciality market, but meanwhile also resis-
tant crops have been developed, which are dis-
tributed in combination with the herbicide (Lib-
erty Link). In 1997 the sales were at $170 ×
106 .
stituted aromatic or heteroaromatic ring system.
The sales of the imidazolinone herbicides grew
with 6.8 % per annum from 1992 to 1997. The
sales for Imazethapyr, the star product against
mono- and dicotyl weeds in soya were about
$540 × 106 in 1997.
Imazaquin [81335-37-7], 2-(4-isopropyl-
4-methyl-5-oxo-4,5-dihydro-1H-imidazol-2-
yl)-3-quinoline carboxylic acid, Scepter,
C17 H17 N3 O3 , M r 311.3, mp 219–224 ◦ C, was
launched in 1984.
Imazapyr [81334-34-1], 2-(4-isopropyl-4-
methyl-5-oxo-4,5-dihydro-1H-imidazol-2-yl)-
nicotinic acid, Arsenal, C13 H15 N3 O3 , M r 261.3,
mp 169–173 ◦ C, was launched in 1985.

The contact herbicide inhibits the plant spe-

cific enzyme glutamine synthetase. The chemi-
cal synthesis requires acrolein, hydrocyanic acid
and methyl phosphinous ester, resulting in a
racemic product [325]. As the l-enantiomer is
Imazamethabenz [81405-85-8], 2-(4-isopro-
40 times more active than the d-compound,
pyl-4-methyl-5-oxo-4,5-dihydro-1H-imidazol-
many asymmetric synthetic routes are under in-
2-yl)-5-methylbenzoic acid methyl ester, As-
vestigation.
sert, C15 H18 N2 O3 , M r 274.3, mp 113–153 ◦ C
(of methyl ester), was launched in 1986.
Bialofos [35597-43-4] [326], Bilanafos,
C11 H22 N3 O6 P, M r 323.3, mp 160 ◦ C (decomp.),
is a natural occurring tripeptide of the sequence
l-phosphinothricine-l-alanine-l-alanine iso-
lated from Streptomyces viridochromogenes,
Streptomyces hygroscopicus and others. The
compound was launched as a nonselective con-
tact herbicide in 1984. It has the same mode of Imazethapyr [81335-77-5], 5-ethyl-2-(4-iso-
action as Glufosinate, but has much lower sales. propyl-4-methyl-5-oxo-4,5-dihydro-1H-imid-
azol-2-yl)nicotinic acid, Pursuit, C15 H19 N3 O3 ,
Herbicides Based on 2-Methylvaline. In M r 289.3, mp 169–173 ◦ C, was launched in
1985, the first member of a new and very im- 1987.
portant class of selective herbicides based on 2-
methylvaline was introduced. These imidazoli-
none herbicides are acetolactate synthase (ALS)
inhibitors [327] meaning that the synthesis of
branched amino acids is blocked [328].
The synthesis of the racemic active ingredi- Imazapic [104098-48-8], 5-methyl-2-(4-iso-
ents starts from methyl isopropyl ketone which is propyl-4-methyl-5-oxo-4,5-dihydro-1H-imid-
converted to 2-methylvaline and finally cyclized azol-2-yl)-nicotinic acid, Cadre, C14 H17 N3 O3 ,

M r 275.3, mp 204–206 ◦ C, was launched in
1997.
Imazamox [114311-32-9], 2-(4-isopro-
pyl-4-methyl-5-oxo-4,5-dihydro-1H-imidazol-
2-yl)-5-methoxymethylnicotinic acid, Raptor,
C15 H19 N3 O4 , M r 305.3, mp, was launched in
1997.
Herbicides derived from Acylphenyl
Amino Acids. Acylphenyl amino acids are a
group of herbicides with decreasing importance.
The development of enantiomerically pure com-
pounds are new concepts for improving the sub-
stance performance.
Benzoylprop [22212-56-2] [329], Suffix,
C16 H13 C12 NO3 (for ethyl ester), has an al-
anine subunit, which is formed by react-
ing 3,4-dichloroaniline with racemic ethyl 2-
chloropropionate and benzoyl chloride. This
herbicide is mainly used in wheat. The racemic
active ingredient and the analogue with an 3-
chloro-2-fluoroaniline aromatic are more and
more replaced by the d-alanine derivative
(racemic switch).
Flamprop-M [63782-90-1] [330], Mataven
L, Suffix BW, C19 H19 CIFNO3 , M r 363.8, mp
72.5–74.5 ◦ C (data for isopropyl ester), is syn-
thesized from 3-chloro-4-fluoroaniline, benzoyl
chloride and methyl S-2-chloro propionate. This
compound is used as selective grass herbicide in
wheat and barley.
Amino Acids 43
Amino Acids in Protox Herbicides. In-
hibitors of the protoporphyrinogene oxidase
[331], also known as Protox, are important her-
bicides blocking the biosynthesis of chlorophyll
in plants. Phenyl substituted heterocycles have a
strong impact on that enzyme [332]. These her-
bicides can contain amino acids in two ways.
On the one side, amino acids like proline, 4-
oxaproline [333] or pipecolinic acid [334] could
be part of the heterocycle forming a hydantoin,
on the other side amino acids like alanine can
form the substituent at 5-position of the aromatic
system. The preemergence activity and selectiv-
ity is increased using the R-enantiomer instead
of the racemic alanine.
Herbicides Based on Glycine. Two herbi-
cides based on a glycine residue are the
chloroacetanilide herbicide Diethatyl [38725-
95-0], Antor, C14 H18 ClNO3 M r 283.75, pro-
duced by Hercules Agrochemicals [318]
and the triazine herbicide Eglinazine [68228-19-
3], C7 H10 ClN5 O2 , M r 231.64, introduced by
Nitrokémia Ipartelepek [318]
5.5.2. Fungicides (→ Fungicides,
Agricultural)
Valinamide Fungicides. A lot of acylvaline
anilides are claimed to be active against a broad
spectrum of fungi, but especially the control of
Plasmopara and Phytophthora [335] is pointed
out. The literature indicates [336] that the S-
compound is the more active enantiomer. This
44 Amino Acids
group of compounds may be divided into valine
derivatives and nonvaline derivatives. The va-
line is usually acylated to form an urethane or a
maleimide. The carboxylic group is condensed
with a substituted aniline, an alkyl benzylamine
or an alkyl homobenzylamine [337, 338].
Two valinamide fungicides have been intro-
duced into the market.
Iprovalicarb [140923-17-7], Melody,
C18 H28 N2 O3 , M r 320.43, has been introduced
by Bayer CropScience for the prevention of
downy mildew and Phomopsis viticola in vines
[339]. The valine moiety is in the l-form, but
the stereochemistry of the second asymmetric
center is not defined.
Benthiavalicarb [413615-35-7],
C15 H18 FN3 O3 S, M r 339.38, has been devel-
oped by Kumiai Chemicals Industry for treat-
ment of tomatoes and potato late blight [340].
Anilide Fungicides. A number of anilide
fungicides with a market volume of approx.
$610 × 106 in 1997 have been established since
the 1990s. Three amino acid derivatives are sold
as fungicides with systemic activity.
Metalaxyl [57837-19-1] [341], Ridomil,
C15 H21 NO4 , M r 279.3, mp 71.8–72.3 ◦ C (data
for isopropyl ester). This most important fungi-
cide of its class is synthesized by condensa-
tion of 2,6-dimethylaniline with methoxyacetyl
chloride and subsequent coupling with racemic
methyl 2-chloropropionate.
This fungicide is mainly used for the seed
treatment business, in wine and in vegetables.
The compound is sold as a stand-alone prod-
uct or in a variety of mixtures with other fungi-
cides. It was introduced in the market in 1978.
The (R)-isomer, based on D-alanine, is known
as Metalaxyl-M [70630-17-0] or Ridomil Gold,
and was introduced by Syngenta in the USA in
1996.
Furalaxyl [57646-30-7], Fongarid,
C17 H19 NO4 , M r 301.3, mp 70 and 84 ◦ C
(dimorphic) (data for isopropyl ester). Fu-
ralaxyl is a condensation product of furan-2-
carboxylic acid and 2,6-dimethylaniline which
is subsequently coupled with methyl 2-
chloropropionate. The compound was intro-
duced in 1984.
Benalaxyl [71626-11-4] [342], Galben,
C20 H23 NO3 , M r 325.4, mp 78–80 ◦ C. In the
commercial synthesis again 2,6-dimethylaniline
is coupled with phenylacetyl chloride and subse-
quently methyl 2-bromopropionate. The single
enantiomer product Benalaxyl-M [98243-83-5],
based on d-alanine, has also been registered.
Hydantoins. Some amino acids being mar-
keted as fungicides are sold as their hydantoins.
One established product is iprodione.
Iprodione [36734-19-7], Rovral,
C13 H13 Cl2 N3 O3 , M r 330.2, mp 134 ◦ C. The
synthesis uses 3,5-dichloroaniline which is cou-
pled with glycine and phosgene. The urea sub-
structure is formed by reaction with isopro-
pylamine and phosgene. Many fungicide mix-
tures with Iprodione are on the market.

Perfuazoate [101903-30-4], Healthied,
C18 H23 N3 O4 , M r 345.4. This rice fungicide has
been introduced by Showa Denko for the treat-
ment of rice seedlings in Japan [318]. It is based
on a racemic homoalanine pentenyl ester back-
bone.
5.5.3. Insecticides (→ Insect Control)
Only a few amino acid based insecticides are
distributed in the market.
tau-Fluvalinate [102851-06-9] [343], Klar-
tan, Mavrik, C26 H22 CIF3 N2 O3 , M r 502.9, bp
164 ◦ C (9.33 Pa) is the most important com-
pound. This substance, preferably used in cot-
ton fields, but also in corn, rape tomatoes and
vegetables, is a pyrethroid with contact and
stomach action. The market of that class had
a volume of $1530 × 106 in 1997. The chem-
ical synthesis starts with 3,4-dichlorotrifluoro-
methylbenzene and d-valine. The carboxylic
acid is esterified with the cyanohydrin of 3-phen-
oxybenzaldehyde. The compound is marketed
with the defined stereochemistry at the valine
moiety, but the cyanohydrin subunit is a R/S mix-
ture.
Imiprothrin, Pralle [72693-72-5],
C17 H22 N2 O4 , M r 318.4, has been introduced
by Sumitomo as a household insecticide against
cockroaches and other crawling insects, and
is a pyrethroid based on glycine hydantoin. It
Amino Acids 45
is sold as a mixture of cis- and trans-isomers
[318]. The long synthetic pathway leads to
the vinyl substituted cyclopropane carboxylate,
which forms an ester with 3-hydroxymethyl hy-
dantoin. The resulting substance is N-alkylated
with 3-bromopropyne.
5.5.4. Plant Growth Regulators (→ Plant
Growth Regulators)
Two compounds with amino acid structure are
highlighted because of their direct chemical re-
lationship to Glyphosate.
Glyphosine [2439-99-8] [344], Polaris,
C4 H11 NO8 P2 was introduced in 1973 and is
preferably used in sugar beets. The synthesis
starting with glycine uses two equivalents of
formaldehyde and of phosphorous trichloride
under oxidative conditions.
A second derivative of Glyphosate is a cyclic
phosphaoxazole being formed from Glyphosate
and formaldehyde. This compound is claimed
to control the growth of some grasses without
discoloration or phytotoxicity.
Aviglycine [49669-74-1], aminoethoxyvin-
ylglycine, ReTain, C6 H12 N2 O3 , M r 160.2, is
produced by Streptomyces spp. and was isolated
and purified by Abbott. It is marketed by Va-
lent Biosciences for control of fruit ripening as
it inhibits the production of endogenous ethyl-
ene [318].
5.6. Industrial Uses
Polyfunctional compounds, such as amino acids
and their derivatives, have been applied in a
46 Amino Acids
wide variety of industrial areas. This is partic-
ularly because of their physicochemical proper-
ties, such as high thermal stability, low volatil-
ity, amphoterism, buffering capacity, and the
ability to form complexes. However, properties
of amino acids such as environmental accept-
ability and low toxicity, are becoming increas-
ingly important. A few of the recorded uses
have been listed below, and include acylamino
acid monomers for epoxy resins [345]; amino
acid dispersing agents for pigments in coloring
polyester fibers [346]; N-acylamino acid dis-
persants for polyurethanes in water [347]; ami-
no acid setting retarders for cement [348]; zinc
salts of N-acyl-derivatives of basic amino acids
and N-acylamino acids for the thermal stabiliza-
tion of PVC [349]; polyglutamic acid esters and
polyaspartic acid esters coatings for natural and
synthetic leather [350]; amino acid hardening
agents for methacrylate resins [351]; N-acylami-
no acid, amino acid ester, and amino acid amide
gel-forming agents for oils [352]; basic amino
acid vulcanization accelerators for natural rub-
ber [353]; amino acid [354] and N-acylamino
acid [355] corrosion inhibitors for metals; ami-
no acids to stabilize the latent image of photo-
graphic emulsions [356]; and amino acid bright-
eners in galvanic baths [357, 358].
6. Chemical Analysis
Amino acids do not have defined melting points
but decompose over a broad range between
250 and 300 ◦ C. Therefore, they must be trans-
formed into derivatives before melting points
are useful for identification. Phenylisothiocy-
anate is used to yield the phenylthiohydan-
toin amino acid (PTH amino acid) [8], or 2,4-
dinitrofluorobenzene (Sanger’s reagent) is used
to yield the dinitrophenyl amino acid [8].
Spectroscopic methods for the identification
of amino acids include infrared [359], Raman,
1
H-NMR, and 13 C-NMR spectroscopy [360] of
free amino acids or PTH derivatives and mass
spectrometry of PTH derivatives [361]. Ultravi-
olet spectroscopy is important only for aromatic
amino acids. The different methods for quali-
tative and quantitative analysis of amino acids,
particularly of mixtures from protein hydrolysis
and biological fluids, have been reviewed [362].
Separation methods are described in [363].
Various staining methods may be used for the
qualitative identification of α-amino acids [364].
Some of these dye-forming reactions are suitable
for quantitative analysis within the validity range
of the Lambert–Beer law. By far the most impor-
tant is the reaction with ninhydrin, which yields
a red-violet to blue-violet dye (λmax = 570 nm).
The imino acids proline and hydroxyproline
form a structurally different dye, with an absorp-
tion maximum at 440 nm.
Fluorescent reagents also have been used
successfully for quantitative analysis. The ami-
no acid is converted into a strongly fluores-
cent derivative, which increases the sensitiv-
ity by orders of magnitude. Typical fluorescent
reagents are o-phthalaldehyde–2-mercaptoetha-
nol [365], 1-dimethylaminonaphthalene-5-sul-
fonyl chloride (dansyl chloride) [366], and
4-phenylspiro [furan-2(3H)-1 -phthalan]-3,3 -
dione (fluorescamine) [367].
The separation of amino acid mixtures is pos-
sible by electrophoresis or chromatography. The
latter is especially useful, techniques including
paper, thin layer, ion-exchange, high-pressure
liquid, and gas chromatography. The different
techniques have been reviewed and compared in
[368]. Paper chromatography is generally car-
ried out in two dimensions. A number of elu-
ents are available for this purpose. Quite often
the amino acid is converted to its dinitrophenyl
derivative.
More common than paper chromatography
is thin layer chromatography (TLC) of either
the free amino acids [369] or their PTH deriva-
tives [370]. Silica gel, aluminum oxide, cellulose
powder, or polyacrylamide may be used as car-
rier, but silica is preferred. An indicator reagent,
most often ninhydrin, is used for detection [371].
For detection of very small quantities the amino

acids separated on an TLC plate can be converted
to the fluorescamine derivatives [372]. The de-
tection limit is 10 pmol. The time required for
the analysis of dinitrophenyl amino acids can be
reduced by using high-pressure thin layer chro-
matography [373]. TLC is often specified as an
analytical test to show the levels of foreign ami-
no acids in a sample, particularly when the ami-
no acid is from natural sources.
Ion-exchange chromatography [374, 375] on
organic resins (Dowex, Amberlite, etc.) has
proven to be the most exact and reliable method
for the separation and quantitative analysis of
amino acids. Before the automatic analysis tech-
nique was introduced [376], complete analysis
of an amino acid mixture required 24 h. To-
day 2 h is the rule. Sodium citrate or lithium
citrate buffer solutions are the eluents. The
eluate is reacted with ninhydrin [374, 377]
or o-phthalaldehyde [378]. With ninhydrin, 1–
20 nmol amino acid can be measured with an
accuracy of ± 1–5 %. The detection limit with
o-phthalaldehyde is in the picomole range. Ion-
pair chromatography on a porous graphitic car-
bon stationary phase has also been applied. De-
tection is by evaporative light scattering, or by
electrospray mass spectrometry [379].
Ion-exchange chromatography is the method
of choice for analyzing amino acids in feeds,
foodstuffs, and biologic fluids [380]. In general,
analysis is preceded by a hydrolysis, which de-
grades the proteins and peptides to their com-
ponent amino acids. However, acidic hydroly-
sis can destroy Cys, Met, and Trp residues, and
special techniques may be necessary to analyze
for these amino acids. Figure 10 shows a sample
aminogram of broiler feed.
Utilization of high-pressure liquid chro-
matography (HPLC) allows a further reduction
in analysis time. Originally, the amino acids
were converted to derivatives, e.g., to dansyl
amino acids [381], PTH amino acids [382],
or dinitrophenyl derivatives [383], before anal-
ysis. Reversed phases are the preferred sta-
tionary phases. Ninhydrin, o-phthalaldehyde–
mercaptoethanol [384], or fluorescamine [385]
are the usual reagents for detection. An HPLC
method with direct UV detection has also been
described [386] for analysis of infusion solu-
tions. Modern automated amino acid analyz-
ers are now available, that use quaternary gra-
dient elution to separate all the common ami-
Amino Acids 47
no acids efficiently without prior derivatization.
These machines generally incorporate postcol-
umn visualization with ninhydrin and photomet-
ric quantification. Today this method rivals ion-
exchange chromatography as the preferred ana-
lytical method.
Gas chromatographic methods [387] are use-
ful for the analysis of complex amino acid
mixtures. However, the amino acids must be
converted into volatile derivatives, e.g., PTH
amino acids [388], methyl esters of N-tri-
fluoroacetylamino acids [389], n-butyl esters
of N-trifluoroacetylamino acids [390], N-tri-
methylsilylamino acids [391], or N,O-bis-tri-
methylsilylamino acids [392]. Gas chromatog-
raphy coupled with electron-capture negative-
ionization mass spectrometry (GC–ECNI–MS)
has been used to determine amino acids at the
femtomole scale, using 2 H or 13 C labeled amino
acids as reference standards [393].
Electrophoresis [394], which employs the
differing rates of migration in an electric field, is
becoming more important [395]. The technique
is already well-established for protein analysis,
and has been coupled with electrospray ioniza-
tion mass spectrometry (CE–ESI–MS) to de-
termine underivatized amino acids at micromo-
lar level [396]. Capillary isotachophoresis is a
new high-resolution electrophoresis technique
for amino acids.
The chromatographic separation of amino
acid enantiomers is the subject of intensive in-
vestigation. Separation is currently possible by
gas chromatography [397] and high-pressure
liquid chromatography [398], using optically ac-
tive phases or chiral solvents [399]. A wide
range of chiral stationary phases is now avail-
able for analytical or preparative separations.
Another method is to use a chiral precolumn
derivatizing agent. Marfey’s reagent (1-fluoro-
2,4-dinitrophenyl-5-l-alanine amide) has been
successfully applied to separate complex mix-
tures of amino acids into their separate enan-
tiomers, and is effective at the nanomolar scale
[400, 401]. Marfey’s reagent forms diastereo-
meric salt pairs with the amino acids, that are
particularly well separated by HPLC.
The microbiological analysis of amino acids
is based on the fact that several l-amino acids are
essential for certain bacteria strains. The growth
of the bacteria cultures under standard condi-
tions can be quantitatively evaluated (acidime-
48 Amino Acids
Figure 10. Amino acid chromatogram of a broiler feed
Internal standard: norleucine. Solid curve: UV detection at λ = 570 nm. Dotted curve: UV detection at λ = 440 nm for Pro (and
Hyp).
try or turbidimetry) and related to the amino acid
concentration. Lactic acid bacteria [402] (Lacto-
bacteriaceae) can be used to analyze 19 l-amino
acids. Typical test microbes include Leuconos-
toc mesenteroides (ATCC 8042), Lactobacillus
arabinosus 17−5 (ATCC 8014), and Streptococ-
cus faecalis R (ATCC 9790, 8043). The conven-
tional microbiological methods are quite com-
plicated but can be simplified by automation
[403].
A series of l- or d-amino acids can be ana-
lyzed by enzymatic methods. l-Amino acids, or
the enantiomeric purity of d-amino acids, can
be determined with bacteria decarboxylases by
measurement of CO2 formed. d-Amino acids, or
the enantiomeric purity of l-amino acids, can
be determined with kidney d-amino acid oxi-
dases by measurement of the O2 consumption.
Enzymes that react only with a single amino
acid allow determination of that amino acid, e.g.,
arginine using liver arginase. An improved en-
zymatic method consists of the use of enzyme
electrodes that contain the enzymes [404], or
microorganisms having a special enzyme in a
fixed form. Enzyme electrodes, however, are rel-
atively unstable [405].
The quantitative determination of crystal-
lized amino acids is carried out by acidimet-
ric titration in nonaqueous medium [406, 407].
Glacial acetic acid is a suitable solvent. Formic
acid may be added to improve solubility. The
titrant is perchloric acid. Formol titration by the
method of Sörensen can be used for aqueous so-
lutions but is less accurate.
Standards of purity for individual l- and d,l-
amino acids that are used in drugs or as food
additives are published in pharmacopeias [406,
408] and food codices [409].
7. Economic Significance
The 2005 world market for amino acids is esti-
mated at more than 3.3 × 106 t/a (Table 19). The
“big three” amino acids, sodium l-glutamate,
d,l-methionine, and l-lysine · HCl, account for
approximately 95 % of the volume (Table 20).
The other amino acids play only a small role.
The dominant amino acid, sodium glutamate, is
used almost exclusively as a taste enhancer. d,l-
Methionine and l-lysine · HCl are used almost
exclusively to improve the nutritive value of an-
imal feeds. l-Threonine is also used mainly in
animal feeds. l-Aspartic acid and l-phenylal-
anine are used principally for the manufacture
of the intense sweetener aspartame. The other
amino acids have diversified applications. With
the exception of glycine, they are more expen-
sive than the big three amino acids. In terms of
 Amino Acids 49

volume, feed additives used about 1 500 000 t China, with established manufacturers setting up
of amino acids, pharmaceuticals 17 000 t and plants there.
sweeteners around 17 000 t (2005) [410]. Ta-
ble 21 shows the market value for amino acids
in 2004 broken down by field of use [52]. The 8. Toxicology
total value of the global amino acids market in
2004 was estimated at $ 4.5 × 109 [52]. Excess amino acids are rapidly disposed of by in-
creased metabolic degradation. Should the ami-
Table 19. Amino acid production, 2005 no acid dose be suddenly increased, e.g., by ex-
Amino acid Quantity, t/a [410–413] tremely high protein consumption, within about
l-Alanine 1500 two days the liver adaptively increases the levels
l-Arginine * 3000 of amino acid-catabolizing enzymes — transam-
l-Asparagine 200
inases, enzymes of the urea cycle, cystathionase,
l-Aspartic acid 15 000
l-Cysteine * 7000 tryptophan pyrrolase, etc. The excess amino
l-Glutamic acid * 1 690 000 acids are to a large extent used to provide energy.
l-Glutamine 2000 The nitrogen is eliminated as urea. A smaller
Glycine 16 000
l-Histidine 300
portion is used in protein synthesis, mainly liver
l-Hydroxyproline 100 protein and plasma albumin.
l-Isoleucine 500 When too little protein or no protein is con-
l-Leucine 800 sumed or when the component amino acids are
l-Lysine * 850 000
d,l-Methionine 600 000 imbalanced, alteration of the ribosome profile
l-Methionine 400 occurs in the liver, and ribonucleic acids are ca-
l-Phenylalanine 14 000 tabolized. The manifestation of chronic protein
l-Proline 800
l-Serine 300
deficiency is known as marasmus (slight defi-
l-Threonine 85 000 ciency) and kwashiorkor (extreme deficiency).
l-Tryptophan 2000 Protein deficiency is usually coupled with calo-
l-Tyrosine 150 rie deficiency (protein–calorie malnutrition).
l-Valine 1100
d-Phenylglycine + 9000
This manifests itself on the biochemical level
d-p-hydroxyphenylglycine as a negative nitrogen balance, indicating a re-
Others 5000 duction in protein inventory. Initially, the labile
Total 3 300 000
enzyme and plasma proteins are consumed, the
* Free amino acid and salts.
* Free amino acid and derivatives. greatest losses occurring first in the liver, then in
the musculature. Brain, heart, and kidneys suf-
Table 20. Percentage of individual amino acids as a part of the fer only minimal protein loss. Symptoms of pro-
total market, 2005 tein synthesis disorders include disturbances in
Amino acid Quantity, % wound healing and bone growth, lowered resis-
l-Glutamic acid (Na) 51
d,l-Methionine 18
tance to infection and stress, loss of fertility and
l-Lysine (HCl) 26 appetite, and anorexia. The urinary excretion of
Other amino acids 5 3-methylhistidine is a common indicator of the
catabolism of muscle protein.
Table 21. Market value by field of use, 2004 The total absence of an essential amino acid
Use Value, % in the diet is more serious than protein defi-
Animal nutrition 56 ciency. In this case the proteins or amino acids
Human nutrition 32 in the diet are totally worthless because protein
Specialty * 12
synthesis can occur only by degradation of body
* Pharmaceuticals, cosmetics, agrochemicals, and industrial uses.
protein. A general interruption of the protein
The main manufacturers of amino acids synthesis results. This manifests itself rapidly
are located in Japan (e.g., Ajinomoto, Ky- as a drop in enzyme activity and an impoverish-
owa Hakko, Tanabe) and in Europe (Degussa, ment of the plasma proteins. Noticeable symp-
Wacker, Amino, Flamma). Recently there has toms are loss of appetite and weight, alteration
been an increasing trend towards production in of cornea and lens, anatomic organ alterations,
and an increased rate of mortality. In addition
50 Amino Acids
there appear specific deficiency symptoms char-
acteristic of the missing amino acid or acids.
The metabolic disturbances brought about by
gross divergences from the optimal amino acid
pattern have three different causes [414, 415]:
imbalance, antagonism, and toxicity.
Amino acid imbalance manifests as an ap-
pearance of deficiency symptoms for the first
limiting amino acid when the other amino acids
are consumed in great excess. The symptoms of
imbalance are eliminated by administration of
the first limiting amino acid in sufficient quan-
tities. The main symptom is a severe reduction
of food or feed assimilation and depression of
growth. The depression of growth in rats has
been investigated intensively by adding individ-
ual l- and d,l-amino acids to basal diets of var-
ious protein levels [416, 417].
Amino acid antagonismis caused by compe-
tition for common transport systems. An exam-
ple is antagonism of the branched-chain ami-
no acids isoleucine, leucine, and valine. The
symptoms are reversible. In a study with young
rats, addition of 5 % l-leucine to a low-protein
diet (9 % casein) reduced the plasma levels of
isoleucine and valine, depressed growth, and re-
duced feed consumption [418]. These effects
were eliminated after a latent period of three
days by small doses of l-isoleucine (0.16 %) and
l-valine (0.15 %).
Amino acid toxicity occurs when very large
quantities of one or more amino acids are con-
sumed and is characterized by total failure of the
adaptive mechanisms. The toxic level has been
studied by adding increasing quantities of indi-
vidual amino acids to a protein basal diet [415].
The toxicity of individual amino acids depends
on the total protein consumption. Imbalance, an-
tagonism, and toxicity are less pronounced when
overall protein consumption is sufficient but be-
come more severe at lower levels of protein con-
sumption. The consumption of toxic amounts of
amino acids increases their concentration in the
plasma and brain. Because of the blood–brain
barrier, however, the increase in the brain is not
as great [419]. Failure of the adaptation mecha-
nisms during consumption of large excess of an
amino acid can lead to accumulation of the ami-
no acids or certain metabolites in the organism,
leading directly or indirectly (e.g., by influenc-
ing hormone secretion) to anatomic or functional
damage.
The toxicity of amino acids has been re-
viewed [415, 418]. The acute toxicities of most
l-amino acids and some derivatives have been
determined [420–422]. The toxicology of d-
amino acids is discussed in review articles [415]
and other publications [421, 422]. There is no
evidence to date that the d-enantiomers of the
α-amino acids found in proteins exhibit specific
toxic effects. Their LD50 values are generally
higher than those of the l-amino acids.
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