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Food fermentations for improved digestibility of plant

foods – an essential ex situ digestion step in agricultural
societies?
Michael G Ga¨ nzle
The fermentation of plant foods detoxifies and eliminates compounds that are inherently present in grains and legumes and have

antinutritive properties, including cyanogenic glycosides, vicine and convicine, phytate, phenolic compounds, immune-reactive

proteins and fermentable oligosaccharides, disaccharides, monosaccharides, and polyols (FODMAPs). Contemporary food

production has partially replaced fermentation of plant foods by alternative processes for production of nutritious plant foods. This

communication explores the question whether the conversion of noxious components in plants by food fermentations remains relevant in

contemporary food production, or, more pointedly worded, whether food fermentations are an essential ex situ digestion step for

agricultural societies. Noxious compounds in grains and legume seeds contribute to irritable bowel syndrome, non-celiac wheat

intolerance, and food allergies. Food fermentations provide an effective unit operation to improve tolerance of plant foods to

sensitive individuals. In addition, they are a source of viable and active microorganisms that may provide additional health benefits.

Address
University of Alberta, Dept. of Agricultural, Food and Nutritional Science, Edmonton, Canada

Corresponding author: Ga¨ nzle, Michael G (mgaenzle@ualberta.ca)

This review comes from a themed issue on Functional foods and

nutrition
Edited by Andreas Schieber

https://doi.org/10.1016/j.cofs.2020.04.002
2214-7993/ã 2020 Elsevier Ltd. All rights reserved.

Introduction
The transition of groups of hunterer-gatherers to agricul- tural societies in the Neolithic Revolution about 14
000 years ago paved the way to modern civilization [1] and substantially impacted the human diet. Hunterer-
gatherer diets included a high proportion of animal protein and diverse plants including fruits, vegetables and tubers. In
contrast, the diet of agricultural societies relied on only few plant species, particularly cereals, legumes, and oil-
seeds. Nutritional consequences of this transition include a reduced intake of protein and dietary fiber, a reduced
supply of minerals, and increased exposure to anti-nutritive factors in seeds [2,3]. The origin of food
fermentations, which are defined as the preparation of foods or beverages by controlled microbial growth and
.. 5.. 6.
enzymatic conversions [4 ], predates the origin of agriculture [ , ]. The case was made that cereals were
..
domesticated to enable fermentation of alcoholic beverages for rituals or festivities about 14 000 years ago [5 ].
While this hypothesis is not undisputed, the knowledge on how to ferment cereals and legumes to improve the
digestibility of grains and legumes, and to remove anti-nutritive compounds likely was a prerequisite for the
Neolithic Revolution. Because fermented foods have unique sensory properties and are often deeply rooted in local
culture, they have remained a staple in the human diet after the shift from artisanal to industrial food
production in the last 150 years.
However, alternative preservation methods including thermal processing or refrigeration, alternative separation
and processing meth- ods including extrusion or isolation of protein fractions, and simplified and accelerated
Current Opinion in Food Science 2020, 32:124–132 www.sciencedirect.com
fermentation methods, for example in production of bread or soy sauce, have reduced the intake of fermented
foods as well as the intake of live and active microbes. This communication explores whether the removal of
anti-nutritive, pro-inflammatory or toxic compounds in cereals grains and legumes by traditional fermentation
processes remains relevant in contemporary human nutrition by improving the tolerance of sensitive individuals to
legumes or cereal products.

Are seeds meant to be eaten?

Toxic, conditionally toxic or anti-nutritive components in grains and legumes
Plants are sessile organisms that cannot escape pathogens or predators and, therefore, employ chemical defenses
against microbial pathogens and herbivores. These chem- ical defenses are termed phytoalexins or phytoanticipins
depending on whether they are pre-formed in the plant (phytoanticipins) or produced after tissue damage
by pathogen or herbivores attack (phytoalexins) [7,8]. Phy- toalexins and phytoanticipins include phenolic
com- pounds and antifungal agents that receive attention as antioxidative food components, food preservatives
9.
and biological fungicides [ ] but also include compounds that have antinutritive properties, or are outright
toxic to humans. Compounds that protect plants against abiotic stress, for example, raffinose-family
oligosaccharides In legumes and fructans in cereals [10,11], also result in adverse digestive symptoms. Toxic,
conditionally toxic or antinutritive food components in plants (Table 1) were identified in the last century and
are described only briefly [12–14].

Cyanogenic glucosides
Cyanogenic glucosides release cyanide after hydrolysis of the bglucosidic bond [14]. Among edible parts of plants,
the content of cyanogenic glucosides is high (0.5– 4 g HCN/kg) in bitter tubers of cassava, lima beans, bitter
almonds, and flaxseed. Chronic cyanide intoxication, leading to a disease termed konzo, is a consequence of
improperly processed cassava and has repeatedly been observed in Africa in times of famine [15]. Comparable
diseases are not reported from South American countries, where cassava originates; this may relate to the long
tradition of safe preparation of cassava roots in South America, which reduces the likelihood of improper prep-
aration even at times when food is scarce.

Vicine, convicine, and favism

Faba beans contain the pyrimidine glucosides vicine and convicine, which constitute up to 2% of the dry weight of
the beans. The aglycones are redox-reactive and have antifungal properties. During digestion, the aglycones are
released through b-glucosidases from faba beans or intes- tinal microorganisms. The aglycones oxidize glutathione in
vivo and cause hemolytic anemia termed favism in individuals with glucose-6-phosphate dehydrogenase
deficiency [16]. Favism is more prevalent in countries where malaria occurs because the resistance of individuals
with low glucose-6-phosphate dehydrogenase activity to malaria provides selective pressure for favism [16].

Phytic acid
Phytic acid is the main storage compound for phospho- rous and minerals in cereal and legume seeds; their
phytate content ranges from 5 to 20 g/kg [13,17]. Phytates are not hydrolyzed in the monogastric digestive tract
2+
until digesta reach the large intestine. The complexes formed by phytic acid and divalent minerals including Ca ,
2+
Zn and iron are insoluble; hence, phytates reduce the bio- availability of minerals. In affluent countries, an
2+ 2+ 2+ 2+
adequate supply of Ca and Zn is warranted by animal products in the diet [18]. Ca and Zn deficiencies
are more commonly observed in developing countries and com- plexation of dietary minerals by phytates in
plant foods contributes to the mineral deficiency [18]. Iron-deficiency anemia is observed in developed as well
as developing countries, particularly in young women [19]. Iron uptake from plant foods is impeded not only by
complexation with phytate but also by complexation with tannins [19,20].
Phenolic compounds
Proanthocyanidins, gallotannins and ellagitannins, com- monly referred to as tannins, are phenolic compounds that
occur in a wide variety of plant foods. Their presence in cereals and legumes is dependent on the plant
species and the cultivar [21]. For example, the content of proanthocyanidins and 3-deoxyanthocyanins in pea
and sorghum varieties, respectively, is highly variable [22,23]. Tannins impart bitter taste, reduce protein and
starch digestibility by inhibition of pancreatic enzymes, and reduce iron uptake [21,23]. In affluent
countries, the presence of tannins reduces the caloric content and the glycemic index of foods [24] while in
countries with low food security, their presence reduces the supply of macro- nutrients and micronutrients.
Enzyme inhibitors
Specific inhibitors of digestive enzymes further reduce the digestibility of starch and proteins in legumes and
cereals. Wheat and other cereals contain amylase-trypsin inhibitors, which account for up to 4% of the total protein
in the grain endosperm [25]. Protease and amylase inhi- bitors are also present in seeds of legumes and oilseeds
[12,21].

www.sciencedirect.com Current Opinion in Food Science 2020, 32:124–132

 Table 1

Toxic, conditionally toxic, antinutritive or noxious compounds in plant foods and milk
Adverse
Adverse compound
effects Food involved
Release
Cyanogenic
of cyanideglycosides
after ingestion; chronic disease
Cassava,
(konzo)flaxseed,
leading to
lima
motor
beans,
neuron
others
damage or acute intoxication [] Favism (hemolytic anaemia) in susceptible individuals with gluco
Bitter taste, inhibition of digestive enzymes [] Faba beans
Inhibition of digestive enzymes, inflammatory effects; potential contribution to non-celiac wheat sensitivity and irritable bowel syndrome []
Vicine, convicine
Allergic reactions, potential anaphylactic shock []
Flatulence, bloating, osmotic diarrhea; contribution to non-celiac wheat sensitivity and irritable bowel syndrome [,]
Lactose intolerance (Flatulence, bloating, osmotic diarrhea) []
Phytate Cereals, legumes
Tannins Sorghum, legumes Wheat, rye, legumes
Amylase trypsin inhibitors, lectins

Allergens Wheat, legumes, eggs, milk

Raffinose-family oligosaccharides; fructans Legumes; wheat, rye
Lactose
Milk

Fermentable oligosaccharides, disaccharides, monosaccharides, and polyols

The content of raffinose-family oligosaccharides in pulses ranges from 1% to 6% with stachyose as most
abundant compound. In cereals, the content ranges from 0.5 to 1.5% and raffinose is the sole or the most
.. 27..
abundant compound [26,27 ]. Cereals, in turn, contain 1–5% of fructans with a DP of 3–6 [ ]. Ingestion of
non-digestible oligosaccharides results in adverse digestive symptoms when a threshold of about 15 g/person
and day is exceeded [28], a threshold that is readily exceeded in cereal-based or legume-based diets unless the
content of these oligosaccharides is reduced by fermentation or germination.

Lactose is not present in plant foods; however, adverse effects of lactose ingestion relate to those caused by
other indigestible oligosaccharides and are thus also briefly discussed in this communication. Infants digest
lactose through the activity of brush-border b-galactosidase (lac- tase); generally, the expression of brush border
..
lactase is reduced after weaning and most human adults do not digest lactose [29 ]. About 25–30% of human
adults are
29..
lactase persistent; the current high prevalence in Euro- pean populations evolved in less than 5000 years [ ].
The ability of humans to digest lactose is thus substan- tially predated by the consumption of fermented dairy
30.
products, which dates back to about 5000 BCE [ ].
Are noxious components of plants and animal foods degraded in food fermentations?
Cyanogenic glycosides
Cyanogenic glycosides are degraded by substrate-derived
b-glucosidase after injury of plant tissue by milling (Figure 1). b-Glucosidase activity of Lactiplantibacillus
..
plantarum (previously Lactobacillus plantarum [31 ]) and other fermentation organisms contributes to the
degradation of cyanogenic glycosides to glucose and the volatile cyanide during fermentation of cassava [32].

Vicine and convicine in beans

Comparable to cyanogenic glycosides, the b-glucosides of vicine and convicine are degraded during fermentation
of
33.
faba beans by substrate-derived or microbial b-glucosi- dases (Figure 1) [ ]. The aglycones divicine and iso-
.
uramil are highly reactive and unstable, and are rapidly removed during fermentation [33 ].

Degradation of phytate
The phytase activity of cereals [13] is sufficient to
35..
degrade phytates if the insoluble salts of phytate and divalent cations are solubilized by acidification [34, ].
Phytate degradation in sourdough thus occurs indepen- dent of microbial phytases. Cereal phytases are optimally
active pH 5.5 but remain active at the lower pH-values that are achieved in sourdough and legume fermentations
35..
[ ]. The use of sourdough in bread production, and lactic fermentations for production of cereal porridges or
beverages thus increase the bioavailability of minerals [36]. The phytase activity in legumes is lower when
compared to cereals [13,37], however, processing of legumes by soaking, milling and lactic fermentation before
cooking substantially reduces phytate levels. One example for such a product is idli, which is produced by
fermentation of rice and legumes in South Asia [38]. Fermented soy products in South East Asia are produced
39.
by steaming or cooking of raw materials before fermentation [ ]. In these products, substrate-derived phytases
are inacti- vated and phytate degradation is achieved by fermenta- tion with bacilli or fungal cultures, for
example, Rhizopus stolonifer or Aspergillus oryzae, which hydrolyze phytate with extracellular enzymes [40,41].
Examples of these products include tempe (or tempeh) produced in Indonesia, stinky tofu produced in China
and natto pro- duced in Japan.
Phenolic compounds
The content of tannins is reduced in fermentations of plant foods [42]. In particular, red and black sorghum
varieties, which are cultivated in sub-Saharan Africa because of their superior drought and pest resistance, are
considered essen- tially inedible unless they are processed by germination and/or fermentation. Only few studies,
however, document
43..
the biochemical conversions of phenolic compounds including tannins at the molecular level [ ,44,45]. Lactic
acid bacteria metabolize tannins by tannases and related phenolic acid esterases (Figure 1) [46]. Tannase
releases gallicacidfromtannins, thusreducingtheir interaction with proteins as well as the affinity to iron. In cereal
fermentations, phenolic acid esterasesof lactobacilli release phenolic acids that are esterified with plant cell wall
polysaccharides [46,47]. Glycosides of phytochemicals including flavonoids are metabolized by diverse glycosyl
hydrolases, releasing the corresponding aglycons [44]. Phenolic acids are metabolized by phenolic acid reductases,
48.. 49..
phenolic acid decarbox- ylases, and vinyl phenol reductases [ , ]. Phenolic acid metabolism by lactic acid
bacteria is strain specific and frequently observed in organisms of the genera Lactiplanti- bacillus, Levilactobacillus and
31..
Furfurilactobacillus (previously L. plantarum, Lactobacillus brevis and Lactobacillus rossiae groups, respectively [ ])
31.. 48..
as well as Limosilactobacillus fermentum (previously Lactobacillus fermentum [ , ], which are dominant fermentation
39.
organisms inspontaneous cereal, legume and vegetable fermentations [ ]. The products of phenolic acid
metabolism include dihydroderivatives of hydroxycinnamic acids, and decarboxylated volatiles which contribute to
the food flavor [46]. Lactic fermentation also generates pyranoanthocyanidins or pyrano-3-deoxyanthocyanidins,
which are formed by conden- sation of vinylphenols, products of decarboxylation of hydroxycinnamic acids by
lactobacilli, and anthocyanidins or 3-deoxyanthocyanidins [45]. The biological activities of the metabolites that are
formed from phenolic compounds
 Nutritional Benefits of Food Fermentations Ga¨ nzle 127

Figure 1

Cyanogenic glucosides

Raffinose-family OS Vicine, convicine

GOS,
Tannins, phenolic acid esters
Flavonoid glucosides

Glu
Fructo-OS Lactose, GOS
Tannases,
Fructans esterase
GtfA

FruA Gal Glu

SucP
Gal Est.
FosE SacA PadR
Phenolic acids or
GshR phenolic acid derivatives
Pad
VprA

Lactate, acetate,
CO2 PtrP
2RSH RSSR

HCN
divicine, isouramil

Flavonoid aglycones
Ptr

Current Opinion in Food Science

Overview on metabolic activities of lactic acid bacteria that contribute to the conversion of anti-nutritive, noxious or toxic compounds in food fermentation.
Fructans. High DP inulin type and levan type fructans are degraded by the cell-wall bound enzymes FosE (Liquorilactobacillus spp. and few strains of L. casei
and L. paracasei) and FruA (oral streptococci and swine-associated Lactobacillus spp.). Fructo-oligosaccharides with a DP of 2 – 4 are metabolized by the
intracellular fructanases SacA and sucrose phosphorylase SucP. Raffinose-family oligosaccharides and galacto-oligosaccharides. Raffinose-family oligosaccharides are
converted to aGOS by extracellular levansucrases (Limosilactobacillus spp.,
Liquorilactobacillus spp. and few Lactobacillus spp.) or metabolized by intracellular a-galactosidase and sucrose-phosphorylase. aGOS and bGOS including lactose
with DP 2 – 4 are metabolized by intracellular a-galactosidase and b-galactosidase, respectively. Cyanogenic glycosides, vicine and convicine are converted by substrate-
derived or intracellular microbial b-glucosidases; the resulting aglycones are rapidly detoxified or
volatile. Phenolic compounds. Flavonoid glucosides are converted by substrate-derived or intracellular microbial b-glucosidases glycosyl hydrolases. Tannins and
esters of phenolic acids are hydrolyzed by extracellular tannases and extracellular or intracellular phenolic acid esterases; phenolic acids are converted by phenolic acid
reductases (HcrB, HcrF or PadR1), decarboxylases (Pad) and vinyl reductases (VprA). Protein modification and hydrolysis. Glutathione reductase activity of F.
sanfranciscensis or other thiol-accumulating enzymes in Limosilactobacillus spp. reduce intramolecular and intermolecular disulfide bonds, increasing the susceptibility
of proteins including ovotransferrin and gluten to hydrolysis. Cell-wall bound proteases of lactic acid bacteria – mainly found in Lactobacillus spp. and lactococci, or
substrate-derived and fungal proteases 27.. 33. 48.. 49..
hydrolyze proteins to peptides and amino acids. Drawn with information from [ ,32, ,44,45, , ,53,82,83,88].

43..
including their nutritional properties, however, remain to be explored [ ].
Degradation of fermentable oligosaccharides, disaccharides, monosaccharides, and polyols FODMAPs including lactose, raffinose-
family oligosac- charides and fructans are partially or completely degraded by fungal or bacterial enzymes during food
fermentations(Figure 1). Fermentation of yoghurt and related fermented dairy products removes only 10–20% of the lactose in
milk[50], however, b-galactosidases of fermentation organisms remain active throughout gastrointestinal transit, hydrolyze lactose, and
thus alleviate lactose intolerance [51]. Raffinose family oligosaccharides are hydrolyzed through the activity of a-galactosidases,
levansucrase and sucrose- phosphorylase activities of lactic acid bacteria [52,53] or corresponding enzymes of fungal cultures; their
removal in legume fermentations has been amply documented (Figure 1) [54]. The removal of fructans in cereal flours has been
explored only recently. In fermentations for bread production, extracellular yeast invertase and intracellular fructanases of lactic
acid bacteria hydrolyze sucrose and low molecu- lar weight oligosaccharides but fructans with a higher degree of polymerization
27..
are not degraded [ ].
The use of lactobacilli or yeasts expressing extracellular fruc- tanases, which is currently employed only in few com- mercial
 27.. 55.
applications, achieves hydrolysis of all fructans for production of low-FODMAP bread [ , ]. Because the adverse effects of
FODMAPs are dose dependent, even their partial reduction in food fermentations alle- viates or even eliminates adverse
symptoms.

Degradation of patulin and other mycotoxins

Patulin is a mycotoxin that is produced by Penicillium expansum and other fungi growing on fruits [56]. Its toxicity relates
to the reactivity with thiols [57], which depletes cellular glutathione levels; accordingly, genera- tion of thiols by yeasts or
heterofermentative lactobacilli during alcoholic or lactic fermentation of fruit juices inactivates patulin. L. plantarum also
converts patulin by uncharacterized esterase and reductase activities [58].
59.
Aflatoxin levels were reduced in cereal and legume fer- mentations [ ], however, pure cultures of lactobacilli do not convert
59.
aflatoxin [ ] and enzyme activities that are known to degrade aflatoxin are not expressed by food- fermenting lactobacilli [60].
59.
It thus remains unknown whether the apparent reduction of aflatoxin levels in food fermentation [ ] is attributable to
absorption of the toxin to bacterial biomass [61], or to the co-operative activity of bacterial and substrate-derived enzymes. It
is thus uncertain whether the reduction of mycotoxins in food fermentations is achieved only by specific combina- tions of raw
materials and fermentation cultures, or relate to a principle that is more generally applicable to fer- mented foods.

Is the fermentative degradation of noxious compounds in plant foods relevant today?

In developed countries with high food security, offending plant foods can be replaced by other plant or animal foods, as is the
case for example, in gluten-free or low-FOD- MAP diets, and food processing provides alternative technologies for
detoxification of plant materials. More- over, antinutritive compounds may be health beneficial in affluent societies, where diets
are often characterized by an excess of rapidly digestible carbohydrates and a low intake of dietary fiber. For example, inhibition
of starch digestion by phenolic compounds decreases the glycemic index and the risk of diabetes and the metabolic syn- drome
[24,28]. Likewise, raffinose-family oligosacchar- ides cause digestive discomfort when consumed in large amounts but have
beneficial prebiotic properties when consumed at an adequate dose [28]. Nutritional benefits of food fermentations, however,
remain relevant even in affluent societies. As outlined further in this section, these nutritional benefits relate to the intake of
viable and non- pathogenic microbes, the reduction of the content of FODMAPs, and modification of immune-reactive pro-
teins in food.

Live and active dietary microbes

Although not all fermented foods contain viable fermen- tation organisms at the time of consumption, fermented
4..
foods are a major contributor to the dietary intake of viable and non-pathogenic microbes [ ,62]. Species that
comprise strains with well documented probiotic proper- ties include Lactobacillus acidophilus, Lactobacillus johnso- nii,
31.. 31..
Lacticaseibacillus casei (previously Lactobacillus casei [ ]), Lacticaseibacillus rhamnosus (previously Lactobacil- lus rhamnosus [ ]),
31..
L. plantarum, Limosilactobacillus reuteri (previously Lactobacillus reuteri [ ]) and L. fermentum are also present in high cell counts
in specific fermented foods [39.,63]. Lactobacilli that are part of fermentation microbiota in fermented foods were shown to exhibit
probiotic properties [4..,64.]. Whether or not probiotic properties should be attributed to fermentation cultures in fermented foods is
discussed controversially [4..,65]; however, the presence of viable and active microbes in fermented foods may address the reduced
intestinal microbial diversity in developed countries.

Fermentation, FODMAPs and irritable bowel syndrome The link of fructans, food fermentations and the irritable bowel syndrome or
non-celiac wheat intolerance has been established relatively recently. Non-celiac wheat intoler- ance is poorly defined and is
66.
diagnosed by elimination of wheat allergies and celiac disease [ ]. Non-celiac wheat intolerance, which affects approximately 6%
of the North American population, largely overlaps with irritable bowel syndrome, which has a prevalence of about 11–
66.
15% [ ]. A majority of individuals with irritable bowel
66.
syndrome or non-wheat intolerance are fructose malab- sorbers [ ]; that is, fructose, which is a digestible sugar
in most individuals, particularly when consumed in asso- ciation with glucose, is non-digestible. A low FODMAP diet, which
necessitates avoidance of wheat and onions, not only improves symptoms of the IBS [67] but also reduces the intake of
dietary fiber and thus reduces the diversity of the intestinal microbiota, particularly by depleting the relative abundance of
bifidobacteria [68]. Health benefits that are associated with ingestion of dietary fiber including non-digestible
69..
oligosaccharides are well documented [28, ]. Low-FODMAP or gluten
free diets may thus have long-term adverse health effects if the ‘fiber gap’ is not addressed.

Does sourdough fermentation improve the tolerance of wheat products for individuals with non-celiac wheat intol-
eranceortheirritablebowelsyndrome? Anecdotalevidence for improved tolerance of sourdough bread is widely shared by sourdough
bakers and social networks, however, scientific evidence for this claim is scarce. FruA mediates degradation of fructans in
27..
sourdough [ ]; this enzyme is generally present inoralstreptococci but present only in few swine-associated lactobacillithatals
27.. 70..
ooccurinone industrial sourdough fermentation [ ,63, ]. FosE, the second extracellular fructanase in lactobacilli, is present
 31..
only in Lactocaseibacillus and Liquorilactobacillus (previously L. casei group and part of the L. salivarius group [ ]), organisms which do
not commonly occur in commercial sourdough
fermentations. Even conventional sourdough fermentation, which is typically carried out with Fructilactobacillus sanfranciscensis
(previously Lactobacillus sanfranciscensis) in combination with Kazachstania humilis (previously Candida humilis) in type I sourdoughs or
Lactobacillus and Limosi- lactobacillus species
31.. in type II sourdoughs (previously L.
delbrueckii group and L. reuteri group [ ,71,72]), reduces the FODMAP content by about 50% through elimination of raffinose family
oligosacchari des and fructans with a low degree of polymerization (Figure 1) [27..,73.]. The improved tolerance of low-FODMAP bread
produced with FruA-expressing lactobacilliin individuals with theirritable
74.
bowel syndrome was demonstrated in clinical trials [ ].Because adverse effects are dose-dependent, however, it is likely that even
conventional sourdough fermentation improves tolerance of wheat and rye bread for a significant portion of susceptible
individuals.

Modification of immune-reactive dietary proteins

The prevalence of allergies in the population of affluent countries is increasing, in parallel with an increased prevalence of
other auto-immune disorders [75]. Fermen- ted foods were associated with a reduced allergenicity when compared to the
corresponding non-fermented counterparts [76]. Fermented foods are not generally hypo-allergenic but specific allergens are
degraded by fermentation of milk [77], soy [78], wheat [79] and eggs [80].
What distinguishes food fermentations from other unit operations in food processing? Fermentation with lactic acid bacteria,
particularly heterofermentative lactic acid bacteria, modifies protein in the fermentation substrate by proteolysis and by
reduction of disulfide bonds (Fig- ure 1). Proteolysis is readily achieved by alternative (enzymatic) processes but sustained
reducing power for modification of disulfide-linked immune-reactive pro- teins requires metabolism by living cells and is achieved
only by germination of seeds or by fermentation. Prote- olysis is a key selection criterion for cultures in dairy fermentations
while substrate-derived or fungal proteases are relevant in cereal and legume fermentations [81,82]. Low-molecular weight thiol
compounds are accumulated particularly by the glutathione reductase and cystathio- nine-g-lyase activity of heterofermentative
lactobacilli including F. sanfranciscensis and L. reuteri [83], key organ- isms in cereal fermentations [71,72]. A specific contribu-
tion of glutathione dehydrogenase of F. sanfranciscensis to protein modification was demonstrated for the hydrolysis of gluten
proteins in wheat sourdoughs [82] and for reduced allergenicity of ovotransferrin [80].

The relevance of protein modifications during food fer- mentation also relates to proteins that are not allergenic but exhibit
pro-inflammatory properties and may exacer- bate or trigger auto-inflammatory diseases. Lectins including the wheat germ
agglutinin are highly disul- fide-bonded and immune-reactive proteins, however, their contribution to adverse health effects in
humans is disputed [84] and their fate in food fermentations is unknown. Pro-inflammatory properties are more clearly
established for the wheat amylase-trypsin inhibitor, which not only inhibits pancreatic enzymes but also induces intestinal
inflammation through activation of Toll-like receptors [85]. The wheat amylase trypsin inhibitor is a hetero-tetramer which
inhibits insect and mammalian amylases [25]. Owing to its pro-inflammatory activity, the wheat amylase-trypsin inhibitor was also
66.
hypothesized to contribute to non-celiac wheat intolerance and the irritable bowel syndrome [ ]. Germination
in combination with sourdough fermentation of wheat flour decreased the trypsin inhibitory activity [86], likely through
degradation of amylase trypsin inhibitors. Sour- dough fermentation of flour from resting grains also reduced ATI levels, and
74.
converted the dominant and biologically active multimeric form to the monomeric form [ ]. However, detailed studies
on the role of proteolysis, reduction of disulfide bonds, heat inactivation during baking, or other factors on the fate of ATI in
(sourdough)-baking are currently lacking.

Food fermentations – an essential ex vivo digestion step for agricultural societies?

Humans have mastered the skill of conversion of agricultural crops by fermentation to improve their nutritional value since the
Neolithic Revolution. Whether the ability to control food fermentations was a prerequisite for this landmark transition in human
history cannot be answered with currently available data; however, several beneficial nutritional aspects of fermented foods
remain relevant at present. Can food fermentations be considered an essen- tial ex situ digestion step to achieve improved
digestibility of plant foods in agricultural societies? This question is partially inspired by the observation that many omnivorous
or herbivorous monogastric animals including swine, poultry and rodents harbor lactobacilli as dominant members of the microbiota
in their upper intestine, that is, the esophagus, the crop and the forestomach [87]. The composition and the metabolic
functions of these animal microbiota substantially overlap with the species level composition and metabolic activity of
lactobacilli in food fermentations [63]. The case can be made that humans compensated the lack of an organ for in vivo
lactic fermentation by using the cognitive function of another organ, the brain, to employ food fermentation as an ex situ
digestion step to improve the nutritional value of plant crops.
Is the case convincing? Probably not. Most humans can digest most plant foods or, in regions with high food
security, are able to substitute offending foods with more appropriate choices.

Is the analogy relevant from a public health perspective? Probably yes. Gastrointestinal disorders including irrita-
ble bowel syndrome, non-celiac wheat intolerance and auto-immune disorders impact a substantial part of the
population in developed countries. Even though an increased consumption of fermented foods may make
only a small and incremental change in these disorders, an increased proportion of fermented foods in the diet, or
a reversion to including more diverse fermentation microbiota, for example, by reinstating sourdough fer-
mentations in bread production, may increase the health and the quality of life in a significant proportion of
the population.

Conflict of interest statement

Nothing declared.

Acknowledgements
The Canada Research Chairs program is acknowledged for financial support. Qing Li and Gautam Gaur are acknowledged for helpful discussions
during preparation of the manuscript.