Enzyme and Microbial Technology 36 (2005) 725728

Production of medium chain glycerides from coconut and palm kernel

fatty acid distillates by lipase-catalyzed reactions
S. Nandi, S. Gangopadhyay, S. Ghosh
Department of Chemical Technology, Oil Technology Division, University of Calcutta, 92, APC Road, Kolkata-700009, West Bengal, India

Received 7 May 2004; accepted 16 December 2004

Abstract

Medium chain glycerides (MCGs) containing C8:0 and C10:0 fatty acids is very much important for medicinal and nutritional applications.
Coconut and palm kernel fatty acid distillates (FADs) can be utilized to produce MCGs by a combination of lipase-catalyzed hydrolysis
and esterification reactions. The neutral glycerides present in coconut and palm kernel FADs are hydrolyzed by Candida rugosa lipase.
The hydrolysates were then subjected to steam distillation under vacuum (at 120140 C) to get fractions rich in medium chain fatty acids
(MCFAs). The fractions, from coconut and palm kernel FADs (75.2 and 76.2% MCFAs, respectively), were esterified with Rhizomucor
miehei (Lipozyme RM IM) lipase to produce MCGs. Products from coconut FAD contained 64.767.5% diacylglycerol (DG), followed by
18.822.9% monoacylglycerol (MG) and 9.89.3% triacylglycerol (TG). Similarly, products from palm kernel FAD contained 63.566.7%
DG, 19.123.6% MG and 9.510.1% TG.
2004 Elsevier Inc. All rights reserved.

Keywords: Fatty acid distillates; Medium chain glycerides; Rhizomucor miehei; Candida rugosa

1. Introduction fective solvent for dissolving cholesterol gallstones in hu-

Medium chain glycerides (MCGs) are mainly utilized as
a nutritional supplement for patients suffering from mal ab-
sorption caused by intestinal resection and also as an infant
feeding formulation [13]. MCGs are useful in treating a
number of medicinal disorders that involve impaired or dam-
aged lipid metabolism, which include obstructive jaundice,
billiary cirrhosis, pancreatitis, cystic fibrosis, celiac disease,
etc. It is also reported to be useful for feeding newborn in-
fants, both to assist their initial growth and contribute to their
physiological development [4].
Medium chain glycerides (MCGs), which are mainly a
mixture of mono-, di- and triglycerides, have specific appli-
cation in the field of foods, pharmaceuticals and cosmetics.
Medium chain monoglycerides are ideal solvent for aromat-
ics, steroids, dyes and perfume bases. A mixture of medium
chain monoglyceride and diglyceride was found to be an ef-
Corresponding author. Tel.: +91 33 2350 8386/6387;
fax: +91 33 2351 9755.
E-mail address: santinathghosh@yahoo.com.hk (S. Ghosh).
mans [5].
The application of lipases for synthetic purpose is now
well documented. Kim and Rhee [5] studied the enzy-
matic synthesis of MCGs by using capric acid and glyc-
erol as substrates in presence of immobilized lipase (Rhi-
zomucor miehei) without any solvent or surfactant. The per-
centage conversion was close to 90% on the basis of capric
acid consumption within 10 h. Ghosh and Bhattacharyya
[6] incorporated 22.125% medium chain fatty acids (MC-
FAs) in coconut oil by lipase-catalyzed polyestermonoester
interchange reaction. Recently Kim et al. [7] incorpo-
rated caprylic acid in perilla oil by two types of en-
zymes, Lipozyme RM-IM and Lipozyme TL-IM at 4851%
level.
During physical refining of fats and oils, fatty acid dis-
tillate is produced as a by-product, which is mainly utilized
in soap manufacturing process. The aim of the present work
is to utilize coconut and palm kernel fatty acid distillates to
produce MCFAs and subsequent production of MCGs by an
enzymatic esterification reaction with glycerol in a solvent
free system.

0141-0229/$ see front matter 2004 Elsevier Inc. All rights reserved.
doi:10.1016/j.enzmictec.2004.12.016
726 S. Nandi et al. / Enzyme and Microbial Technology 36 (2005) 725728

2. Materials and methods HP make. The oven temperature was programmed from 100
to 190 C at 5 /min. The injector, and detector block tem-
2.1. Materials peratures were maintained at 230 and 240 C, respectively.
IOLAR-2 nitrogen was used as the carrier gas (flow rate
Coconut and palm kernel fatty acid distillates were ob- 30 ml/min). The fatty acid ester peaks were identified and
tained from Edible Products (India) Pvt. Ltd., Kolkata. Glyc- calibrated with standard methyl esters. Data are averages of
erol (A.R.) was purchased from E. Merck (India) Pvt. Lim- three determinations.
ited. Amano 30 (Candida rugosa) was a kind gift of
AMANO ENZYME Inc., Nagoya, Japan. The lipase NS
40013 (Candida antartica, a non-specific, immobilized li- 3. Results and discussion
pase) is also a kind gift of Novozymes, South Asia Pvt. Ltd.,
Table 1 shows the total fatty acid composition, unsaponifi-
Bangalore, India.
able matter and neutral glycerides present in coconut and
2.2. Enzymatic hydrolysis palm kernel fatty acid distillates (FADs). The neutral glyc-
erides present in the FADs are monoacylglycerol (MG), di-
Each fatty acid distillate (100 g) was taken in a 250 ml acylglycerol (DG) and triacylglycerol (TG) of which TG is
stoppered Erlenmeyer flask, and water (60% by weight of the major component. The total fatty acid composition of the
neutral glycerides) containing 1.0% Amano 30 lipase pow- coconut and palm kernel FADs is represented in Table 2. Co-
der. The reaction mixture was magnetically stirred with a 1 in. conut and palm kernel FADs contain total (sum of C8:0 and
Teflon coated stir bar at 35 2 C in a controlled water bath. C10:0 ) of 13.1 and 21.9% MCFAs, respectively.
The degree of hydrolysis was determined by the content of The neutral glycerides present in FADs are hydrolyzed to
free fatty acid in the sample periodically withdrawn. After free fatty acids (FFA) by Candida rugosa (Amano 30) li-
complete reaction, the fatty layer and water layer containing pase powder. The hydrolysis reaction was investigated as a
enzyme and glycerol were separated by centrifugation. function of time as shown in Fig. 1. Within 6 h the neutral
glycerides were more or less completely hydrolyzed. The hy-
2.3. Fractional distillation drolyzed FADs are then fractionally distilled under vacuum to
get a fraction rich in MCFAs. Table 3 shows the yield and fatty
The hydrolyzed fatty acids were subjected to fractional acid composition of the different fractions and residues. The
distillation in a Claisen-Vigroux (250 ml) flask. The flask was fraction collected at 120140 C both from coconut (Fraction
connected to a vacuum pump through a distillation head, an I) and palm kernel (Fraction III) FADs is rich in MCFAs. The
air condenser and a previously weighed 100 ml round bottom yield of Fraction I from coconut FAD is 15.4% which con-
collecting flask. The fractions were collected at 120140 C tains 75.2% MCFA (29.2% caprylic, C8:0 and 46.0% capric
and 140160 C at 4 mmHg pressure for about 30 min. The C10:0 ). The fraction collected from palm kernel FAD as Frac-
feed, distillate and residue fractions were weighed and ana- tion III (yield 14.2%) contains 76.2% MCFAs (47.2% C8:0
lyzed. and 29.0% C10:0 ).
Fractions I and III containing a significant amount of MC-
2.4. Enzymatic esterication reaction FAs are then esterified with glycerol using non-specific NS
The distilled fatty acid and glycerol in an appropriate pro-
Table 1
portion were taken in a round bottom flask and stirred by a Analytical characteristics of coconut and palm kernel fatty acid distillates
magnetic stirrer at 60 2 C for 8 h using 10% (by weight (FADs)
of substrates) NS 40013 lipase. The stopper of the flask was Properties Coconut FAD Palm kernel
kept open to eliminate the water formed during esterification (%, w/w) FAD (%, w/w)
reaction. The esterification reaction was monitored by esti- Free fatty acid (as lauric acid) 71.3 61.2
mating the free fatty acid content in the samples periodically Unsaponifiable matter 0.5 1.2
withdrawn. After complete reaction, the product mixture was Neutral glycerides
isolated and mono-, di- and triglycerides, and their amounts Monoacylglycerol (MG) 2.4 1.4
were determined by column chromatographic method. Diacylglycerol (DG) 2.5 1.9
Triacylglycerol (TG) 23.3 34.3
2.5. Gas chromatographic analysis
Table 2
Fatty acid composition was determined by a gasliquid Total fatty acid composition of coconut and palm kernel fatty acid distillates
chromatographic (GLC) method after converting the fatty Fatty acids (%, w/w)
acids into methyl esters. The HP-5890A GLC was connected
8:0 10:0 12:0 14:0 16:0 18:0 18:1 18:2
with a HP-3390A data integrator. The GLC was fitted with
a glass column (1.83 m 3.175 mm i.d.), packed with 10% Coconut 4.5 8.6 45.5 17.2 9.9 1.4 10.8 2.1
Palm kernel 11.2 10.7 38.6 24.1 7.7 1.6 4.3 1.8
DEGS supported on Chromosorb-WHP (100/200 mesh), of
 S. Nandi et al. / Enzyme and Microbial Technology 36 (2005) 725728 727

Table 3
Yield and fatty acid composition of the fractions by fractional distillation of hydrolyzed coconut and palm kernel FAD
Yield (%) % Fatty acid composition
8:0 10:0 12:0 14:0 16:0 18:0 18:1 18:2 MCFA
Coconut
Fraction I 15.4 29.2 46.0 22.7 1.7 0.4 75.2
Fraction II 34.2 0.0 3.4 66.8 14.6 5.5 0.6 7.5 1.6 3.4
Residue 50.4 0.0 1.2 36.6 23.6 16.2 2.5 2.6 17.3 1.2
Palm kernel
Fraction III 14.2 47.2 29.0 22.4 0.7 0.2 0.4 0.1 0.0 76.2
Fraction IV 39.8 1.8 6.9 63.2 20.0 3.1 0.5 3.9 0.6 8.7
Residue 46 0.4 0.5 19.1 21.9 24.9 9.4 9.1 0.9
MCFA: medium chain fatty acids (8:0, 10:0).

Fig. 1. Biohydrolysis of coconut and palm kernel fatty acid distillates

(FADs). Enzyme: crude lipase: Amano 30 (Candida rugosa); tempera-
ture: 35 2 C.
40013 lipase to get MCGs. Figs. 2 and 3 demonstrate the time
course of enzymatic esterification reaction with MCFAs pro-
duced from coconut and palm kernel FADs, respectively. The
esterification reactions almost come to an equilibrium after
8 h. Fatty acid and glycerol concentration was maintained at
2:1 and 2:1.2 molar ratio just to produce more and more MG
and DG in the finished product and also to reduce the free
fatty acid content at lower level within a reasonable time pe-
riod. The composition of the esterified products is shown in
Table 4. Both products I and II from coconut FAD contained
a significant amount of DG (about 64.767.0%), followed
by MG (18.822.9%) and TG (9.89.3%). Similarly, prod-
ucts III and IV from palm kernel FAD contained 63.566.7%
Fig. 3. Enzymatic glycerolysis of the distilled fatty acids from palm kernel
FAD. Enzyme used: non-specific lipase NS 40013 (Candida antartica); ratio
of fatty acid to glycerol was 2:1 and 2:1.2 for products III and IV, respectively;
temperature: 60 2 C.
DG, 19.123.6% MG and 9.510.1% TG. With the increase
of glycerol content in the reaction mixture the equilibrium
was little bit shifted towards the MG. The products can be
used as MCGs for food and pharmaceutical purpose.
In conclusion, the valuable MCFAs can be produced from
coconut and palm kernel FADs. MCFAs are readily converted
into MCGs by lipase-catalyzed esterification reaction. Mi-
crobial lipase technology may be suitable in producing better
quality of the concerned products from the relatively inferior
grade raw material.

Table 4
Composition of the medium chain glycerides (MCGs) from coconut and
palm kernel FAD
Composition (%, w/w)

FFA% MG DG TG
Coconut
Product I 3.9 18.8 67.5 9.8
Product II 3.1 22.9 64.7 9.3
Fig. 2. Enzymatic glycerolysis of the distilled fatty acids from coconut FAD. Palm kernel
Enzyme used: non-specific lipase NS 40013 (Candida antartica); ratio of Product III 4.1 19.1 66.7 10.1
fatty acid to glycerol was 2:1 and 2:1.2 for products I and II, respectively; Product IV 3.4 23.6 63.5 9.5
temperature: 60 2 C.
728 S. Nandi et al. / Enzyme and Microbial Technology 36 (2005) 725728
Acknowledgements
The research was financially supported by a research grant
from Department of Biotechnology, Government of India.
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