Available online at www.sciencedirect.
com
Traditional biotechnology for new foods and beverages
Jeroen Hugenholtz
The food and beverage industry is re-discovering fermentation
as a crucial step in product innovation. Fermentation can
provide various benefits such as unique flavor, health and
nutrition, texture and safety (shelf life), while maintaining a
100% natural label. In this review several examples are
presented on how fermentation is used to replace, modify or
improve current, artificially produced, foods and beverages
and how also fermentation can be used for completely novel
consumer products.
Address
Swammerdam Institute for Life Sciences, University of Amsterdam and
Coca-Cola Corporate Research, Mainburger Strasse 19, 84072 Au/
Hallertau, Germany
Corresponding author: Hugenholtz, Jeroen (jhugenholtz@coca-
cola.com)
Current Opinion in Biotechnology 2013, 24:155–159
This review comes from a themed issue on Food biotechnology
Edited by Elaine E Vaughan and Jeroen Hugenholtz
For a complete overview see the Issue and the Editorial
Available online 8th February 2013
0958-1669/$ – see front matter, # 2013 Elsevier Ltd. All rights
reserved.
http://dx.doi.org/10.1016/j.copbio.2013.01.001
Introduction
The process of fermentation was traditionally leveraged
by the human population to increase the shelf life of
perishable agricultural produce such as milk, vegetables
and meat. This has resulted in a vast variety of fermented
foods and beverages that is still the main part of the
human diet in many under-developed countries and in
most countries in South-East Asia. In North America and
Europe which have extremely efficient and rapid distri-
bution systems and an overall availability of cooling and
freezing systems, most of the traditional fermented pro-
ducts, with the exception of fermented dairy (yoghurt and
cheese) and meat (sausages) have been replaced by fresh
agricultural produce, making the process of fermentation
obsolete. This trend away from fermentation seems to
have come to a recent stop and is now gradually reversing.
More and more people (and food companies) are regain-
ing interest in traditional and more natural foods and
there is a growing dislike for the processing and energy-
input that is needed to maintain freshness of agricultural
crops. In addition, the food and beverage industry is
continuously trying to innovate within the constraints
of sustainability and naturalness. All these recent devel-
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opments are leading to increased interest and activity in
fermentation technology by all consumer goods indus-
tries, big and small. This trend is not really visible in the
scientific literature and (not yet) in the patent literature,
so some referencing will be made to company-websites as
support of statements made in this contribution.
Fermentation for all-natural foods or
beverages
Fermentation, nowadays, is all about bringing unique
signature flavors and other benefits to consumer products
in a 100% natural way. This is done using, mainly, two
different approaches. The ‘easiest’ is to turn a traditional,
home-grown, product into a large scale process. Examples
are the dairy product, kefir [1], which is a traditional liquid
fermented dairy product using a mixture of lactic acid
bacteria, yeast and fungi for the fermentation of milk,
resulting in a yoghurt-like, slightly alcoholic product
which has traditionally been consumed by millions of
people in, especially, Eastern Europe and supposedly
conveys spectacular health benefits to the consumer
[2]. The fermentation process is, typically, conducted
at home on, or close to, the stove and using small left-
overs of previously produced kefir or a small kefir granule
purchased at the local grocery store, as inoculum. Only
recently, the larger dairy companies have managed to
upscale this process to an industrial scale using active and
stable starter cultures just as is the daily practice for
mainstream dairy products such as yoghurt and cheese.
Another example is the traditional drink of Kvass (http://
rbth.ru/articles/2012/06/22/kvass_russias_real_national_-
drink_15952.html), which is a result of fermentation of
kitchen left-overs mainly consisting of bread bits. This
fermented product is also traditionally made at home with
a final composition depending on the type of left-overs
used and the nature of the locally evolved microbial
culture. This traditional beverage is now also marketed
by larger companies, also outside the originating country,
Russia. The main challenge of these companies is to
prepare this family drink with a constant quality and,
especially, without alcohol. This is basically done by
using controlled fermentations with standardized starter
cultures and similar fermentation conditions (short, low
temperature) as used for non-alcoholic beer production.
Several more examples can be found of food and beverage
companies launching new or improved fermented
products based on traditional biotechnology [3]
(http://bycristinaybarra.com/2012/10/27/burnarj-the-first-
sparkling-drink-with-andalusian-taste/, http://www.calpis.
net/features/story/index.html, www.the-spirit-of-georgia.
de, http://www.doehler.com/en/our_products/beverage_
and_dairy_bases/fermented_products) (Figure 1).
Current Opinion in Biotechnology 2013, 24:155–159
156 Food biotechnology
Figure 1
Kombucha
o d n t al
pr me tion
uc ed
fe adi
ts
Tr
r
Lassi
Kvass
Example of traditional fermented beverages from across the world.
A second approach followed by many consumer goods
companies is to transfer a successful, large-scale, fermen-
tation process to other substrates. One typical example is,
again, kefir, which traditionally is a dairy product, using
milk as substrate. The newest development is to use the
traditional kefir culture for fermentation of other sub-
strates, such as fruit juice, resulting in products called
‘fruit kefir’ or something similar. Other examples are
biological soft drinks such as Bionade (http://dizzyfrinks.
com/drink/bionade-holunder-elderberry/) (in Germany,
Switzerland, Austria) and Tumult (http://news.softpedia.
com/news/Coca-Cola-Comes-Out-with-Tumult-the-Soft-
Drink-that-Tastes-like-Beer-216150.shtml) (in France)
where the microorganisms of the traditional, fermented
tea drink Kombucha [3] have been used for fermentation of
malt, leading to a non-alcoholic soft drink with unique
flavor for the adult market.
In addition to offering unique sensory sensations, as the
label on the package or the advertisements will most likely
tell us, several other benefits can be offered using the
process of fermentation. In this overview only some recent
developments in the health arena will be discussed, such as
natural fortification with vitamins, benefits addressing
obesity and increase of anti-oxidant content.
Fermentation for natural fortification
Over the past 10 years, the process of fermentation has
become recognized as a relative easy and one hundred
Current Opinion in Biotechnology 2013, 24:155–159
Kefiy
Boza Amazake
Amasi Fruit vinegars
Current Opinion in Biotechnology
percent natural technology to raise vitamin levels in foods
and beverages. Numerous examples, especially involving
dairy applications, have been presented showing natural
enrichment with riboflavin [4], folate [5], vitamin B12 [6],
vitamin K2 [7] and sometimes several of these vitamins,
simultaneously [8,9]. The latest developments in this area
are discussed in another chapter of this Special issue [10].
On the basis of the successful applications in dairy products
[4–6], vitamin enrichment in many other fermented food
products should be possible. Using the high folate-produ-
cing Lactobacillus plantarum strain described by Hugen-
schmidt and co-workers [11], many novel fermented
consumer products seem possible since many traditional
fermented products involve the use of Lb. plantarum and
this lactic acid bacterium can be found on almost any
agricultural crop including animal produce. Vitamin for-
tification has been described for a large number of
traditional fermented foods such as soy sauce [12], kimchi
[13], and several others [14]. The work by Santos and co-
workers, just show that this process could also be used for
novel fermented consumer products, in this case fermen-
ted melon, where both the lactic acid bacteria Lactobacillus
reuteri and Lb. plantarum showed much higher folate pro-
duction on melon than on other natural substrates [9].
Fermentation for addressing obesity
The developed world is facing a major threat in the
overall health of its population. In many countries,
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 Traditional biotechnology for new foods and beverages Hugenholtz 157
already more than half of the population is overweight
and the number is growing at an alarming rate. These
overweight people do not just lack discipline in that they
eat too much and have too little exercise, but often have
reached a serious state of illness (obesity) that needs
medical attention. Food and beverage industries (are
trying to) show their responsibility in these health issues
and are putting major R&D efforts in developing con-
sumer products that lead to reduced overall food and
energy intake. One focus of this R&D is to reduce
carbohydrate calories in food and beverages. Fermenta-
tion, almost by definition, will contribute to calorie
reduction by converting sugars into organic acids or
ethanol, although the maximum calorie reduction that
can be reached in such fermentation processes cannot be
more than 25%, since organic acids contain 3 kcal/g versus
4 kcal/g for sugar. Another approach of the food and
beverage industry is to develop consumer products or
ingredients that lead to early satiation of the consumer.
The satiation factors can convey their activity at different
moments in the sequence of eating and digestion, from
sensing the product before consumption, to physically
bulking the stomach or by release of signals at a later
location in the digestive tract. One of the signals that have
been shown to play a role in this satiety cascade, is organic
acids such as propionic acid, butyric and acetic acid
[15,16,17]. These studies indicate that short chain fatty
acids regulate food intake through control of gut hormone
expression such as peptide PYY and glucagon-like pep-
tide. For acetic acid (28 mmol in a serving of vinegar)
[18,19] and propionic acid (2.45% in bread) [20] actual
dosages have been described that lead to satiety response
in humans through delay of the gastric emptying rate and
lowered blood glucose and insulin reponses. As these
acids are typically produced by fermentation, satiety-
inducing fermented foods or beverages could be pro-
duced using a propionic, acetic and/or butyric acid bac-
terium as a starter. In South-East Asia, and especially
Japan, products from acetic acid fermentation, such as
vinegar, have a traditional healthy image, and several
other health benefits, such as protection against inflam-
matory bowel diseases and colon cancer, seem to be
associated with the fermentative production of short
chain fatty acids [21,22].
Fermentation for increasing anti-oxidant
activity
Many fruits and vegetables are more-and-more appreci-
ated for their natural content of valuable anti-oxidants,
such as lycopene (tomatoes), equol (soy), resveratrol (red
grapes, berries, pomegranate), hesperidin and naringin
(oranges), and quercetin (fruits, vegetables). Besides pro-
tecting the fruit or fruit juice directly from oxidation and
undesirable browning and flavor changes, these anti-
oxidants are also, sometimes, claimed to convey anti-
oxidant activity to the consumer and protect the human
cells from destruction by oxidative radicals. The efficacy
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of these anti-oxidants is mostly determined by how they
are taken-up by the human body. Recent studies have
shown that enzymatic and microbial activity (fermenta-
tion) can lead to conversions of several polyphenolic anti-
oxidants such as hesperidin [23,24] and equol [25,26],
leading to more efficient uptake and thus higher bioavail-
ability of the plant polyphenols. As such, fermentation
could also contribute to improved health of the consumer
on the level of increased anti-oxidants and there are
clinical studies indicating that this is indeed the case
for the examples of orange (hesperidin) and soy (equol)
[24,26]. For a more detailed overview on polyphenols in
foods and their supposed health aspects, the reader is
referred to another chapter in this Food Biotechnology
issue [27].
Fermentation for even longer shelf-lives
Traditionally fermented consumer products have rela-
tively long shelf-lives ranging from about 4 weeks for
liquid fermented dairy products to a year or longer for
some dry fermented sausages and alcoholic beverages.
Still, there is always a need for longer and more complete
protection. The fermented foods and the fermented, non-
alcoholic, beverages usually still contain large residual
levels of sugar and of various amino acids, vitamins and
minerals allowing growth of a large number of acid-
tolerant microorganisms which are mainly yeast or fungi.
To address this problem, food microbiology research has
devoted much attention to finding microorganisms, and
especially lactic acid bacteria, that produce antifungal
compounds. A number of antifungal-producers have been
identified [28–30] and also some unique antifungal com-
ponents have been elucidated [31,32] such as various
cyclic peptides and metabolites such as 3-phenyllactic
acid. A recent study by Crowley and co-workers show that
such antifungal-producers can also be used for longer
shelf-lives of fresh fruits such as pear and grapes [33].
When these antifungal-producers were applied by us in
various fruit juices, the (fermented) products, clearly,
showed longer protection against fungal growth
(Figure 2). The challenge for the food industry is, now,
to make sure that these antifungal fermentations still
have the right flavor attributes, that they combine well
with the fermentations that deliver other benefits and that
the right strategy is chosen with respect to labeling
legislation (f.i. in situ fermentation for flavor and ot-
her — antimicrobial — benefits will lead to other labeling
than addition of fermented ingredients with the direct
purpose of controlling fungal growth).
Conclusions and perspectives
Fermentation can deliver many benefits to foods and
beverages. Besides unique flavors and textures as exem-
plified in traditional fermented products such as yoghurt,
cheese, soy sauce and kimchi, many novel benefits,
especially with respect to health, can be conveyed via
fermentation technology. All this can be achieved via
Current Opinion in Biotechnology 2013, 24:155–159
158 Food biotechnology
Figure 2
(a) (b) (c) (d)
Current Opinion in Biotechnology
Effect of lactic acid bacterial fermentation on fungal growth in pear juice.
The pear juice fermentations with Weillonella confuse strain 7 (a),
Lactobacillus plantarum strain 62 (b) and Lactobacillus plantarum strain
16 (c) were conducted at 308C for 48 hours before the, forced, fungal
infection was applied. A spontaneous fungal-infected pear juice was
used for infection. A non-fermented pear juice (d) was also infected with
the same fungi, as a control Subsequently, incubation was continued for
3 more days. The fermentation with strain Lp16 shows clear anti-fungal
protection. The three lactic acid bacteria were selected from the study of
Crowley et al. [33].
complete sustainable and 100% natural processes either
by up-scaling existing, traditional kitchen-style processes
or by using successful fermentation processes/microor-
ganisms on novel substrates.
With the use of novel genomic technologies
more detailed insight has been given in some
traditional fermentations such as sourdough [34], kefir
[1], kimchi [35,36], cacao [37], yoghurt [38] and sau-
sage [39] fermentations. Also several lesser-known fer-
mented products have been characterized using these
genomics tools [40,41]. It has clarified for instance
which microorganisms are most active in these fermen-
tations and how they are adapted to their specific
environments. This type of experimental data is crucial
when considering to up-scale these processes or when
applying these microorganisms on other substrates.
Such data will also be used to build comprehensive
models of each fermentation process with the ultimate
goal to accurately predict the outcome of (novel) fer-
mentations. The struggles and challenges faced by the
modelers are discussed in another chapter of this
special Food Biotechnology issue [42]. However, some
early successes of the Systems Biology approach
are also reported [43,44,45] providing possible
solutions for process improvement and also providing
the essential insight for much more extensive use of the
fermentation technology in the food and beverage
industry.
Current Opinion in Biotechnology 2013, 24:155–159
References and recommended reading
Papers of particular interest, published within the period of review,
have been highlighted as:
 of special interest
 of outstanding interest
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A very nice example of how genomics is used for discovery of novel
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Current Opinion in Biotechnology 2013, 24:155–159