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Natto, a traditional soy food fermented by Bacillus subtilis (natto), is made by steaming or cooking soaked soybean seeds, inoculating them with the bacteria, and then letting them sit for an incubation period. Natto soya has grown in popularity because of its nutritional importance and health advantages. As a result, farmers have more opportunities thanks to the natto soybean market. For the natto soybean market to remain stable and grow, improved soybean cultivars with enhanced natto quality traits are essential. Natto's high-quality attributes are influenced by the bacteria strain, processing parameters, and soybean variety. Natto has a specific flavour and aroma with a slimy, sticky consistency. Natto contains a range of essential nutrients and bioactive compounds, i.e. nattokinase, soybean isoflavone, γ-polyglutamic acid, vitamin K2 and biogenic amines. Natto possesses various therapeutics. Bacterial species, processing conditions, and cultivars of soybean determine the quality characteristics of natto. Natto food is higher in menaquinone-7 and contains 100 times more menaquinone-7 than most cheeses. The present review highlights the production technology, microbiology, nutritional composition, and therapeutic potential of natto.
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Review Article
Nutritional Health Perspective of Natto: A Critical Review
Muhammad Afzaal ,
Farhan Saeed ,
Fakhar Islam ,
Huda Ateeq ,
Aasma Asghar ,
Yasir Abbas Shah ,
Chigozie E. Ofoedu ,
and James S. Chacha
Department of Food Science, Government College University, Faisalabad, Pakistan
Department of Home Economics, Government College University, Faisalabad, Pakistan
Department of Food Science and Technology, School of Engineering and Engineering Technology,
Federal University of Technology, Owerri, Imo State, Nigeria
Department of Food Science and Agroprocessing, Sokoine University of Agriculture, P.O. Box 3006, Chuo Kikuu,
Morogoro, Tanzania
Correspondence should be addressed to Muhammad Afzaal; and James S. Chacha;
Received 1 June 2022; Revised 25 July 2022; Accepted 8 September 2022; Published 21 October 2022
Academic Editor: Saleh Ahmed Mohamed
Copyright ©2022 Muhammad Afzaal et al. is is an open access article distributed under the Creative Commons Attribution
License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is
properly cited.
Natto, a traditional soy food fermented by Bacillus subtilis, is made by steaming or cooking soaked soybean seeds, inoculating
them with the bacteria, and then letting them sit for an incubation period. Natto soya has grown popular because of its nutritional
importance and health advantages. As a result, farmers have more opportunities, thanks to the natto soybean market. For the natto
soybean market to remain stable and grow, improved soybean cultivars with enhanced natto quality traits are essential. Natto’s
high-quality attributes are influenced by the bacteria strain, processing parameters, and soybean variety. Natto has a specific flavor
and aroma with a slimy, sticky consistency. Natto possesses various therapeutic potentials and contains a range of essential
nutrients and bioactive compounds, i.e., nattokinase, soybean isoflavone, c-polyglutamic acid, vitamin K
, and biogenic amines.
Bacterial species, processing conditions, and cultivars of soybean determine the quality characteristics of natto. Natto food is
higher in menaquinone-7 and contains 100 times more menaquinone-7 than most cheeses. e present review highlights the
production technology, microbiology, nutritional composition, and therapeutic potentials of natto.
1. Introduction
Natto is a fermented soybean food that was introduced
thousands of years ago in North Japan [1]. ere are three
classes of natto, such as hamanatto, itohiki, and daitokuji
natto. Itohiki natto is a kind of natto that has been inoculated
with bacteria and cultured for 24 hours without the addition
of salt. Daitokuji, or hamanatto, is prepared by injecting it
with mold, raising it for 4–6 months, and adding salt to it [2].
Many states in Asia even have an equivalent product to
natto, which includes the Philippines, Korea, ailand, and
China. Natto has 59% moisture, 16% protein, and 10% lipid
in common; natto was produced from bristled soybeans and
infused with Grass bacillus [3]. Soybean fermentation by
Grass bacillus will create a mildewed flavor, an oily
appearance, and a specific odor produced by sticky and
viscous polymers [1]. Several compounds, including gluta-
mic acid, amino acid, and fructan, are present in the sticky
polymer. In Japan, natto was prepared with mustard sea-
weed, finely sliced onion, and a minor quantity of soy, so it
was always presented with steaming rice [3]. Besides being a
low-priced healthy food consumed with a pair of chopsticks,
natto can also find application as a potential food mix in
various food products. ey should be coated with a white-
colored slimy material that has a distinct flavor, a light
yellow hue, and is able to produce a silky and sticky mass
with a palatably soft texture. Furthermore, natto is used to
flavor fish, meat, and vegetables [4]. Denaturation of soy
proteins through heat, trypsin inhibitors, and bacterial-
enzymatic protein degradation into simply digestible
Biochemistry Research International
Volume 2022, Article ID 5863887, 9 pages
peptides boosts the nutritional value during the production
process. After enough fermentation, the undamaged soy-
beans are covered in a white-colored sticky fluid and have a
softer texture, a slimy apperance, and a distinct flavor [5].
Fermented natto products are also eaten without cooking
and can be stored in cooler places or freezers, even within the
supermarket, for selling and purchasing. Natto can be served
with a couple of other food ingredients; for example, in
Japanese homes, traditional natto is presented with seaweed,
thinly chopped onion, mustard, and a small amount of
condiment, and together with steamed rice, it is served as an
entremet [6]. Natto can also be used for the preparation of
meat, vegetable, and seafood dishes as a flavoring agent as
well as an ingredient for the production of sauce [7]. Soy-
bean fermentation produces proteolysis activity, which
improves the taste and nutritional value by removing un-
wanted flavors [8]. Owing to the increase of isoflavone
aglycone, the fermentation of soybean has an antidiabetic
effect. To check the effects of fermentation time on total
bioactive content and antioxidant activity, different time
practices were made [9]. Additionally, different ratio
practices have been done previosuly by incorporating Sac-
charomyces cerevisiaewhich affected the sensory character-
istics and pH of black soybean natto. However, during the
Taisho Period (1912–1926), scientists developed a method to
synthesize Bacillus natto in the laboratory without the use of
a straw. When placed in pots of cooked soybeans, the new
laboratory-grown bacteria act as a dependable starter cul-
ture, allowing the production of natto [10]. Natto could
subsequently be produced effectively by utilizing industrial-
scale equipment such as big steaming/boiling caldrons. After
adding the appropriate number of bacteria, the steamed
beans might develop in any clean, non-corrosive container.
Several biological events were reported during soybean
fermentation by Hayashi et al. [11]. e amount and rate of
the reactions and material formed are based on the situation
of steaming, soaking, bacteria straining, and fermentation.
Natto has an unusual flavor, smell, consistency, thickness,
and quantity of mucus under different fusions of processing.
Within this study, there are not even any small-level research
laboratory techniques available to assess the soybean cul-
tivar's acceptability for natto processing. Nowadays, black
beans are solely used to make soy sauce, but black soybeans
have been proven to be medicinally useful, with extracts used
as an anti-inflammatory agent [12]. According to the most
recent data, more than 700,000 tons of natto are produced
each year. e present review highlights the therapeutic
potential, production method, nutritional composition, and
microbiology of natto in detail.
2. Production Technology of Natto
Natto is made from soybeans by the action ofBacillus subtilis
var. natto (also known as Bacillus natto). Soybeans are a
catch-all term for both black and green soybeans. Apart from
the main components that separate soy proteins, fats, car-
bohydrates, cellulose, ash content, and moisture, there are
also various trace elements and vitamins. Bacillus natto is a
bacterium separated from traditional Japanese food, and its
first type is similar to Bacillus subtilis, a subspecies of Bacillus
subtilis [13]. Traditionally, it is wrapped in soybeans and
stored in a warm area for 1–2 days. Computer network
control provides a range of effective controls that may in-
crease stable product quality, minimize utility consumption,
and reduce production costs (Figure 1). ey may also be
used to manually participate in and monitor CR production,
reduce quality risk, and reduce production costs.
3. Microflora and Microbiology
Natto production includes the following steps: washing and
soaking whole grain soybeans overnight in hot water. e
seeds are then cooked for 20–30 minutes at 0.98–1.47 Bar
vapor pressure. Cooked beans are refrigerated to 45°C before
being innoculated with a probiotic bacterial strain, and then
natto is fermented for 18–20 hours at 40–45°C [15].
B. subtilis is a Gram-positive, fast-growing, aerobic bacte-
rium with rod-shaped cells usually 2–6 μm long and im-
partial below 1 μm wide. e optimal temperature is at
30–35°C, which allows for twice as much time as 20 minutes.
Under certain growth conditions, the cells form long chains
that connect to unspecified septal wall components. Under
starving conditions, cells can undergo a complicated two-cell
division that results in endospore creation; this formation is
discharged by lysis of the covering mother cell [16]. In other
words, they can produce biofilms and “fruit bodies” con-
taining grains. Isolated in the 1950s, tryptophan auxotroph
was the most popular and studied type of B. subtilis 168. It
was the first known gene to complete a genetic sequence,
displaying a 4.2 Mbp chromosome containing 4100 genes.
With a series of updates, the B. subtilis genome still has one
of the best annotations. However, recently, “SubtiWiki” (a
complete database) provides an easy-to-use and authentic
configuration for the latest data (Table 1). Nicolas’ work has
resulted in a comprehensive data set for writing values,
facilitators, and RNA controls on the website [18]. A
complete list of key genes has been found in a number of
global projects, recently identifying genes (the 257 genes)
needed for Lactobacillus growth at 37°C(total genetics) of
approximately 6250 genes and the “core genome” (genetic
unit) of approximately 2500 genes. Considerable genetics
involves a number of genes, about 300 genes were needed to
build the endospore, as well as many prophages or fossils of
the phage. e conclusions from the appearance of genetic
matter are reconcilable with the idea that B. subtilis is
modified for life in plants and the rhizosphere by Nicolas
et al. [17].
4. Nutritional Composition
Natto provides 211 calories per 100 g. One serving contains
19 g of protein, 11 g of fat, and 13 g of sugar. e leftover
carbohydrate complex contains 5.4 g of fiber and 4.9 g of
sugar. Natto includes 1.6 g of saturated fat and provides
13.0 mg of vitamin C, 8.60 mg of iron, 729 mg of potassium,
and 217 mg of calcium per 100 g. erefore, natto is a food
that falls under the category of “legumes and legume
products.” Natto has a variety of carbohydrates. Each kind
2Biochemistry Research International
has its own range of advantages. One cup of natto contains
around 6 grams of natural sugar. Another type of carbo-
hydrate present in natto is fiber, Anderson et al. [19]. When
you utilize a full cup of cooked food, you will obtain more
than 9 grams of fiber. Adults should ingest 28 g of fiber per
day, according to the USDA. Eating fiber not only helps
digestion and exercise but it also has other health advan-
tages, such as a lower risk of several cancers, obesity, heart
disease, and diabetes [20]. One cup of natto has more than
19 g of fat. e majority of these lipids are polyunsaturated.
Polyunsaturated fats can decrease LDL cholesterol and may
reduce the risk of heart disease and stroke. Natto is a protein-
rich food, so if we increase our plant-based protein intake by
34 grams while utilizing a big cup, natto contains an
abundant source of micronutrients. e natto supplement
gives 2.7 mg of manganese, a total of 130 to 134% of your
recommended daily diet. It provides about 1 mg of copper
(58% of your daily needs), 15 grams of iron (84% of your
daily needs), 1276 mg of potassium (36%), 201mg of
magnesium (50%), 305 mg of phosphorus (30%), 5.3 mg of
zinc (35%), and 15.4 mcg of selenium (22%). Food is high in
ascorbic acid, which provides about 23 mg or about 38% of
your daily requirements. e byproduct of the millet pro-
cessing industry, millet bran, is rich in nutrients, particularly
dietary fiber [21].
Natto contains vitamins and other
important compounds that helps to boost the immune
system [22]. e food industry concentrates on using bio-
active compounds because of the growing interest in doing
so to maintain product quality and safety, as well as the
benefits they have for human health and the environment
[23]. Next, a discussion of applications and functionalization
strategies for the administration of therapeutics via various
delivery methods [24]. Natto has numerous health benefits,
including the ability to control blood cholesterol levels,
Soaking 10 – 12 h
1.5 kg/cm2, 20 min
103 spores/g soybean
40°C, 16 – 18 h
Below 10°C, 24 h
Spore inoculation
Cooling and Aging
Figure 1: e mechanism of natto production is depicted in the flow sheet. Source: [14].
Table 1: Important microorganisms and enzymes used in food fermentation.
Microorganisms Species Active enzymes References
Yeast Saccharomyces cerevisiae Amylase, proteases, maltase, dehydrogenase βglucosidase, and alcohol Nicolas et al. [17]
Bacteria Bacillus subtilis Peptidase, hydrolases, proteases, amylase, and cellulase Nicolas et al. [17]
Biochemistry Research International 3
prevent arterial sclerosis, heart disease, and hypertension,
promote bone growth, control the bacterial balance in the
intestines, prevent diarrhea, enteritis, and constipation,
improve immunity, fat reduction, beauty treatment, eye
relief, and so on[25]. Finally, natto is frequently mentioned
as one of the greatest sources of vitamin K, particularly
vitamin K
. Vitamin K is used by the body to create bone and
prevent blood clottingaccording to Anderson et al. [19].
Several studies on the health benefits of natto have been
conducted; furthermore, in vivo and in vitro studies have
demonstrated that serine proteases such as subtilisin and
nattokinase have a profibrinolytic effect [26–28]. Mamiya
and Nishimura [29] found that rats fed with natto had
enhanced locomotor activity. Natto has some amazing
benefits for bone development in menopausal women and
postmenopausal bone loss prevention, which is most likely
due to the presence of menaquinones or non-nato flavones
in natto [30, 31].
5. Therapeutic Potential
5.1. Anticarcinogenic Activity. Natto has anticancer prop-
erties. A good example is miso soup, a well-known tra-
ditional Japanese dish that is prepared with soybeans as a
major ingredient. is soup is basically made from the
paste of soybeans that have been cooked with mold, yeast,
and bacteria before being blended with water and salt. In
order to prepare 200 ml of miso soup, commercially
available natto in a quantity of about 50 g was added and
cooked for 1 minute. All the volunteers ate miso soup daily
at mealtime [32]. Human gastric adenocarcinoma cells
were used to study the anticancer properties of chung-
kukjang (a Korean short-term fermented soy paste), and
Bacillus strains from chungkukjang were isolated and
identified. K-Chungkukjang (87%) demonstrated the
strongest growth inhibitory effect at a concentration of
1 mg/mL, followed by H-chungkukjang (85%) and MC-
chungkukjang (69%) (P<0.05) reported by Seo et al. [33].
As per epidemiological studies, high levels of isoflavonoids
are particularly related to a decreased colon cancer risk,
while miso soup intake is linked to a lower risk of stomach
cancer. Beans have been reported to hold large amounts of
carcino-preventive agents. Bowel cancer was not that
common in Japan, but it has now become more common
among Japanese people due to the high-fat consumption in
modern Japanese food, Adlercreutz [34].
5.2. Antibacterial Spectrum. e microbial cytotoxicity of
polyphenols may result from nonspecific interactions with
polysaccharides, inhibition of proteolytic enzymes (pepti-
dases), as well as other interactions that render bacterial
adhesins deactivated [35]. Natto (B. subtilis) possesses an-
tibacterial properties in terms of Escherichia coli O157.
Growth of E. coli O157 in culture and natto (B. subtilis)
growth were both reduced. Natto promotes the growth of
Lactobacillus,Bacillus, and Streptococcus while inhibiting the
growth of E. coli in rat caeca reported by Sumi [36]. Natto’s
commercial diet contained a kind of Bacillus that might be
beneficial as a biocontrol agent (Figure 2). Under optimal
circumstances, natto displayed a substantially stronger in-
hibitory activity against S. aureus. erefore, Bacillus subtilis
has antibacterial action. Under well-designed circumstances,
B. subtilis natto had the strongest activity in preventing
S. aureus. Bactericidal action against Helicobacter pylori has
been found in Bacillus subtilis natto. Because of the presence
of dipicolinic acid, a small test of inhibitory concentration
indicated that B. subtilis natto possessed antiplatelet ag-
gregation and anti-H. pylori properties [39].
5.3. Effect on the Immune System. ere has recently been a
lot of excitement concerning the potential of immuno-
therapeutics. Tumor vaccines and immune checkpoint in-
hibitors are two immunotherapy strategies that have
demonstrated a great promise in both clinical and preclinical
trials [40]. Rivera-Patron et al. [41] investigated the effects of
Bacillus subtilis natto on body function in dairy calves during
the feeding period. Bacillus subtilis natto was mixed with
milk and administered orally to calves. When the calves’
initial diet reaches 2% of their body weight, they are fed.
Bacillus subtilis natto enhanced working conditions by
improving daily gain and nutritional efficiency, as well as
increasing the weaning age of calves. In Bacillus subtilis,
there were no increases in serum IgM, IgA, or IgE, but serum
IgG was greater in natto-supplemented calves than in
control calves studied by Rivera-Patron et al. [41].
5.4. Anti-Inflammatory and Hypocholesterolemic Effects.
Soy protein with an eye or an eye alone has been demon-
strated to decrease oxidative stress and have anti-inflam-
matory characteristics by decreasing nuclear factor-kappa B
(NF–B) and preventing the release of chemical cytokines.
e anticancer effect of lunasin (a peptide present in soy-
beans) comes from natto, which has antioxidative and anti-
inflammatory properties. Lunasin, an antioxidant, was
shown to slow down 2,20-azino-bis (3-ethylbenzothiazoline-
6-sulfonic acid) diammonium salts and inhibit inflamma-
tory cytokines (TNF- and IL-6) in RAW mouse 264.7
macrophages, Smithson et al. [42]. In general, soy iso-
flavones inhibit mTORC1 via the AKT pathway, which
causes a decrease in lipogenesis and adipogenesis and an
increase in lipolysis and oxidation in DIO male rats. is
study shows soy isoflavones have amazing impacts on body
weight and adiposity [43].
5.5. B-Galactosidase Activity. Subtilis bacterial cells (con-
taining pNC61AV10 or pNZ2) were deposited in trimeth-
oprim 10 milliliters of LactobacillusLactobacillus medium,
and by the absorption rate of 600 nm, growth was examined.
All the samples were mixed in, and the pellets were im-
mersed in Z buffer (7 ml solution) (pH 7, 40 mM
0, 60 mM Na
0, 10 mM KCl,
4Biochemistry Research International
1 mM phenyl methyl fluoro sulfate, 1 mM MgSO
and 2-mercaptoethanol). e cell suspension of 2.1 millili-
ters has three drops of toluene, and the solution was
forcefully extracted for 9–10s. en, to evaporate the tolu-
ene, the samples were stirred for 1 h at 37°C and collected at
28°C for 5 minutes. e reaction mixture was immediately
stirred for 1 minute at 28°C, after adding 0.6 ml of ortho-
nitrophenyl galactoside solution (pH 7, 4 mg of orthoni-
trophenyl galactoside per ml in 0.1 M sodium phosphate
buffer). e reaction was stopped by adding 1 M Na
(1.5ml) to the mixture. After the cells were discharged for 2
minutes, the absorption of clear supernatant (at 420 nm) was
observed by Wang and Doi [44].
5.6. Gastrointestinal Proliferation. e use of extracts from
certain Bacillus strains in natto, such as targeted microbial
(direct-fed microbial), has been proven to have the ability to
germinate and enter the digestive systems of diverse animals,
such as poultry [45]. As a result, they are active and give a
variety of nutritional advantages, such as the creation of
exogenous enzymes such as cellulose, protease, phytase,
lipase, keratinase, and xylanase, as well as other chemical
compounds that improve the regulation of the body. In vitro,
Bacillus spp. Xylanase was selected by using the in vitro
digestive model. Cellulase production as DFM was then
tested for digestion viscosity and C. perfringens to increase
the diversity of chicken feed. e results of this study
revealed that the use of less expensive grains (fermented
soybean products) for poultry feed improved the digestion
and activity level of birds found by Hendricks et al. [46].
5.7. Antidiabetic Effect. Hyperlipidemia and obesity are
frequently linked with type II diabetes and insulin resistance,
both of which lead to metabolic illness. Remarkably, fer-
mented soybeans have antidiabetic and hypolipidemic
properties in animals [47]. Soybean products that are
fermented and contain soy protein, such as Bacillus subtilis
natto, have been shown to be particularly helpful in reducing
type 2 diabetes in humans. For six weeks, women aged 19–39
years were given a diet that included soy protein (20–30%
other plant protein, 30–35% animal protein, and 30–35% soy
protein). Jiang et al. [48] did an experiment and fed the rat
soy protein-supplemented sucrose-rich foods. e results
showed that the rat reduced cholesterol and hepatic tri-
glyceride storage, steatosis, normal glucose-6-phosphate,
and glycogen levels, and glucose transporter GLUT4
transplant. When this supplement is given for 4 weeks to
diabetic Wister rats, it increases insulin sensitivity, insulin
signaling, and pancreatic function studied by Kwon et al.
5.8. Antiallergic Properties. An immune hypersensitivity
disorder called allergy is caused by an allergy that enters the
body through skin contact, injection, ingestion, and/or
smell. ese immune responses can develop into allergies,
including inflammation such as atopic dermatitis, asthma,
anaphylaxis, food allergies, and allergen rhinitis [50]. Lee
et al. [51] found that natto has antiallergic effects on epi-
dermis pigmentation, ear thinning, internal lymph nodes,
and mast cell infiltration, among other things.
5.9. Antioxidative Properties. Many lifestyle-related disor-
ders are induced by free radical oxidative damage to the
living body. Reactive oxygen species and antioxidant
mechanisms are out of balance, which leads to oxidative
stress [52]. e study found that fermented-soy-products
boosted antioxidant profile, total phenolic content, total
flavonoid content, and isoflavones content. e variations in
isoflavones might be attributed to -glycosidase activity [53].
Natto, or cooked beans, is a traditional Japanese meal that
has been consumed for many years. Antioxidant activity has
been demonstrated in the soluble components of natto
Natto food
glutamic acid
K2(MK7) Nattokinase
B. subtilis
Figure 2: Potential effects of Bacillus subtilis natto. Source: [37, 38].
Biochemistry Research International 5
water. Iwai et al. [54] have also reported that natto fractions
suppress plasma low-density lipoproteins. LDL oxidation is
recognized to play a role in the genesis and progression of
arteriosclerosis. In this study, hypercholesterolemic mice fed
a meal containing one or two natto components, a low
molecular weight viscous or a soybean water extract, were
found to have a considerable effect on LDL oxidation in
vitro. To explore the inhibitory impact of natto fractions on
LDL oxidation in vivo, lipid peroxidation in plasma and LDL
were assessed following natto-treated mouse’s plasma oxi-
dation [54].
5.10. Effect on the Blood Pressure Level. Natto is a soybean
derivative that is a popular traditional meal in Japan and is
also used as a health supplement. Omura et al. [55] dis-
covered that NKCP®, a natto supplement derived from the
enzyme bacillopeptidase F, has antithrombotic, fibrinolytic,
and antihypertensive properties. e utilization of dietary
supplements present in traditional Japanese food provides
further advantages in removing the independent symptoms
of patients getting health treatment who have life-threat-
ening diseases [56].
5.11. Protection against Apoptosis. Apoptosis is a hereditary
cell death formation that plays an important function in cell
number control. e diminished capacity to trigger apoptosis,
which is coupled with changes in cell growth control systems,
has a significant pathogenic feature in many forms of cancer
[57]. e results from this study showed that L. acidophilus
improved apoptosis in treated mice and reduced the severity
of colorectal carcinogenesis. One of the most dangerous
malignant epidermal cancers is melanoma. e natto, or
soybeans fermented by Bacillus subtilis natto, was used to
isolate natto freeze-drying extract (NFDE) and natto water
extract (NWE), which were evaluated as potential anti-
melanoma agents (Table 2). Cell cytotoxicity tests showed that
NFDE and NWE had significant, dose-dependent anti-
melanoma effects while having little effect on normal skin
cells such as Hs68, HaCaT, and adipose tissue-derived stem
cells (ADSCs) reported by Chou et al. [58].
6. Future Perspective
e development of food science in the near future probably
depends on the continuation of active food science, an idea
that was first introduced in Japan about 15 years ago. Japan,
however, followed a unique path of progress in a product-
driven environment rather than a science-driven science. In
fact, the number of substances and products that have the
potential to decrease the risk of disease rather than simply
for health care has been investigated by their body-changing
functions [59]. Some of them have been used in the man-
ufacture of processed foods in accordance with the “defined
health food use” officially defined by the new law. Probiotics
have GRAS certification and are widely ingested across the
world without any issues with safety. It has been demon-
strated in a number of in vivo and in vitro studies. However,
recent studies have revealed concerns about their safety and
their capacity for immunocompetent individuals, Redman
et al. [60]. However, probiotics are very healthy to promote
health, especially in the prevention and treatment of diar-
rhea as well as H. pylori infection and maintaining intestinal
homeostasis [61]. eir use with certain antibodies, espe-
cially for those who are severely ill, newborns, and the el-
derly, should be carefully monitored since reports of
bacteremia in immune-prone patients treated with spores
and other probiotics [62]. On the other hand, the impor-
tance of identification at the stage of severity is also sig-
nificant to detect and eliminate any fundamental
communication between probiotics and problems separated
by allergies involved in the immune system. erefore, it is
essential to remember that clinical experiments involving
these building materials should include a sufficient number
of targeted individuals, including people with low immune
systems. However, there is a dire need to conduct more
clinical studies to investigate the therapeutic potential of
7. Conclusion
Natto is a fermented soybean product that has a unique
microbial profile and abundant bioactive compounds. e
results of various studies conducted on natto suggest that
natto has high probiotic potential. Significant research has
been conducted to improve B. subtilis natto strains, with a
focus on boosting the synthesis of useful chemicals such as
nattokinase, PGA, and isoflavones, among others, hence
improving the health benefits of natto soybeans. As a
result, combining breeding soybean varieties with
B. subtilis natto strains will allow for greater natto health
promotion. ere is strong evidence in the literature that
B. subtilis natto has a high potential for producing pro-
biotics for use in human food, and natto intake has been
linked to health advantages such as a decreased incidence
of certain illnesses and a lower risk of death.
Data Availability
e data that support the findings of this study are available
from the corresponding author upon request.
Table 2: Lactic acid bacterial strains and their function.
erapeutic property Bacterial strains Regulatory chemicals References
Antioxidant damage DNA L. acidophilus and L. Casei 5-fluorouracil Elmore and Susan, [57].
Immune boosting L. acidophilus DCs cell Elmore and Susan, [57].
cytokines IL-12 and IL-10
Epigenetics Lipoteichoic acid and L. acidophilus expression of tumor suppressor genes Elmore and Susan, [57].
6Biochemistry Research International
Conflicts of Interest
e authors declare that they have no conflicts of interest.
e authors are thankful to the Government College Uni-
versity, Faisalabad, for the literature collection facilities.
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... To address this gap and simultaneously facilitate the production of validated 'fermented foods' by manufacturers and the consumption and diffusion of fermented foods in the public, we establish a model approach for assessing and augmenting the microbial content of common fermented foods. The goal is to provide food manufacturers with a cost-effective and powerful tool to integrate the already positive characteristics of most fermented foods [30,36,[46][47][48][49] with the specific health-promoting attributes of certified probiotic strains. Not only will this allow manufacturers to better design experiments and clinical trials to leverage 'probiotic' labels, but it will likely lead to more health benefits for consumers. ...
... The ability to identify multiple probiotic strains that can be used in a given fermented food can facilitate the production of multi-strain probiotic fermented products that also fulfill the health goal of maintaining and increasing gut microbial diversity [4,[56][57][58]. For example, the cardiovascular health benefit associated with Bacillus subtilis in natto [49,59,60] can be combined with the cholesterol-lowering effect of Lactobacillus acidophilus, Bifidobacterium lactis, Lactobacillus plantarum or Lactobacillus reuteri [61,62] by screening for their ability to survive and grow together in the specific conditions dictated by natto ingredients and fermentation conditions. Using the approach demonstrated here, our overall goal is to stimulate the application of this model in the large-scale production of probiotic fermented products characterized by different thermal treatments, sources of raw material, and aging conditions typical of many commercial fermented foods already largely known and widely consumed (Kefir, cheeses, labne, feta, sauerkraut, pickles, olives, kimchi, gochujang, ssamjang, doenjang, natto, miso, etc.). ...
Full-text available
The past few decades have demonstrated how important the human gut microbiota is for human health. Because of this, the use of microbiota-modulating dietary interventions such as probiotics and prebiotics is growing in popularity amongst consumers, food manufacturers, healthcare professionals, and regulators. In particular, there is interest in making a wider variety of foods with probiotic properties. However, as a solution for food manufacturers to produce fermented foods compatible with the “probiotic foods” label definition, we used an impedometric analysis to identify the survival and growth capacity of microbial strains in specific environmental contexts. Using this approach, manufacturers can more effectively select the strains with the highest growth rate for use in probiotic fermented food production trials. To provide a proof of concept, we tested three Lacticaseibacillus rhamnosus probiotic strains growing in milk at different temperatures. We quantified the probiotic’s growth using species-specific primers and quantitative real-time PCR. Overall, our results demonstrate the feasibility of this type of model in facilitating the production of probiotic fermented foods by allowing manufacturers to select strains able to grow under specific conditions. Our model can be used to develop, increase, and target the beneficial health properties of a multitude of fermented foods produced worldwide.
... Previous studies on spontaneously fermented soybean varieties of southeast Asian countries (Cheonggukjang, Natto, Thua nao and Pe poke) and India (Kinema) revealed a colossal diversity of Bacillus sp. (Afzaal et al. 2022;Kharnaior and Tamang 2022;Tamang et al. 2021Tamang et al. , 2022Wongsurawat et al. 2023 ...
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Preparation of traditionally fermented soybeans varies across ethnicities with distinct tastes, flavour, and nutritional values. The fermented soybean varieties Hawaijar, Bekang, and Akhone of north-east India are associated with diverse ethnic groups from Manipur, Mizoram, and Nagaland, respectively. These varieties differ in substrate and traditional practice that exerts differential bacterial-metabolite profile, which needs an in-depth analysis i. Culture-dependent and independent techniques investigated the bacterial diversity of the fermented soybean varieties. Gas chromatography and mass spectroscopy (GC-MS) studied these varieties’ metabolite profiles. The common dominant bacterial genera detected in Hawaijar, Bekang, and Akhone were Bacillus, Ignatzschinaria, and Corynebacterium, with the presence of Brevibacillus and Staphylococcus exclusively in Hawaijar and Oceanobacillus in Bekang and Akhone. The metabolite analysis identified a higher abundance of essential amino acids, amino and nucleotide sugars, and vitamins in Hawaijar, short-chain fatty acids in Bekang, polyunsaturated fatty acids in Akhone and Hawaijar, and prebiotics in Akhone. The bacteria-metabolite correlation analysis predicted four distinct bacterial clusters associated with the differential synthesis of the functional metabolites. While B. subtilis is ubiquitous, cluster-1 comprised B. thermoamylovorans/B. amyloliquefaciens, cluster-2 comprised B. tropicus, cluster-3 comprised B. megaterium/B. borstelensis, and cluster-4 comprised B. rugosus. To the best of our knowledge, this is the first comparative study on traditional fermented soybean varieties of north-east India linking bacterial-metabolite profiles which may help in designing starters for desired functionalities in the future.
... (Isnes-Gembloux, Belgium). The selection of the probiotic bacteria used in the cocktails was mainly based on their potential health benefits widely reported in the literature [30][31][32][33][34][35]. In addition, genomic sequencing and identification of metabolites was conducted by the suppliers. ...
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Bacteriotherapy represents an attractive approach for both prophylaxis and treatment of human diseases. However, combining probiotic bacteria in “cocktails” is underexplored, despite its potential as an alternative multi-target therapy. Herein, three-strain probiotic mixtures containing different combinations of Bacillus (Bc.) coagulans [ATB-BCS-042], Levilactobacillus (Lv.) brevis [THT 0303101], Lacticaseibacillus (Lc.) paracasei [THT 031901], Bacillus subtilis subsp. natto [ATB-BSN-049], Enterococcus faecium [ATB-EFM-030], and Bifidobacterium (Bf.) animalis subsp. lactis [THT 010802] were prepared. Four cocktails (PA: Bc. coagulans + Lv. brevis + Lc. paracasei, PB: Bc. subtilis subsp. natto + Lv. brevis + Lc. paracasei, PC: E. faecium + Lv. brevis + Lc. paracasei, PD: Bc. coagulans + Lv. brevis + Bf. animalis subsp. lactis) were tested using a short-term (72 h) simulation of the human colonic microbiota in a final dose of 6 × 10⁹ CFU. All these probiotic mixtures significantly increased butyrate production compared to the parallel control experiment. PA and PB promoted a bifidogenic effect and facilitated lactobacilli colonization. Furthermore, reporter gene assays using the AhR_HT29-Lucia cell line revealed that fermentation supernatants from PA and PB notably induced AhR transactivity. Subsequent examination of the metabolic outputs of PA and PB in intestinal epithelial models using cell culture inserts suggested no significant impact on the transepithelial electrical resistance (TEER). Assessment of the expression of proinflammatory and anti-inflammatory cytokines, as well as AhR-related target genes in the Caco-2 cell monolayers indicated that PB’s metabolic output upregulated most of the measured endpoints. This in vitro investigation evaluated the potential impact of four multispecies probiotic mixtures in the human colonic microbiota and identified a promising formulation comprising a combination of Bc. subtilis subsp. natto, Lv. brevis, and Lc. paracasei as a promising formulation for further study.
... Animal studies and human trials demonstrate that natto possesses anti-thrombotic and anti-coagulant properties. Furthermore, natto exhibits anticarcinogenic activity, antibacterial spectrum, antiinflammatory and hypocholesterolemic effects, b-galactosidase activity, anti-diabetic effect, antiallergic properties, antioxidative properties, apoptosis protection, and many more (Afzaal et al., 2022;Chan et al., 2021). ...
... An innovative technology (IT) based on conventional methods (CM) incorporating steam pretreatment and illumination fermentation was applied to produce HJTF of better flavor. Steam pretreatment of raw materials is conducted to prepare several traditional soy foods, such as douchi (Zhang, Li, et al., 2022), meju (Kim, Han, Baek, Chun, & Jeon, 2022), and natto (Afzaal et al., 2022). Raw materials pretreated with heated water and/or high-pressure steam are easier to ferment and have improved flavor. ...
... An innovative technology (IT) based on conventional methods (CM) incorporating steam pretreatment and illumination fermentation was applied to produce HJTF of better flavor. Steam pretreatment of raw materials is conducted to prepare several traditional soy foods, such as douchi (Zhang, Li, et al., 2022), meju (Kim, Han, Baek, Chun, & Jeon, 2022), and natto (Afzaal et al., 2022). Raw materials pretreated with heated water and/or high-pressure steam are easier to ferment and have improved flavor. ...
... Therefore, it is still unclear what mechanism can be associated with the contribution of natto to health . A pack of natto contains about 19 g of protein (Afzaal et al., 2022). Since the general recommended daily allowance (RDA) of protein for adults is 0.8 g/kg/day, so eating natto at three meals a day can be considered to provide the required human protein, suggesting that natto may be a useful food source of protein (Trumbo et al., 2002). ...
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Natto, fermented food produced by Bacillus subtilis natto (B. subtilis natto), is widely consumed around the world, and its potential health functions are attracting attention. Natto has been reported to have a variety of bioactive compounds such as levan, menaquinone‐7, nattokinase, and others. Such bioactive compounds produced by B. subtilis natto are deeply involved in the function of natto, because they are consequently contained in natto. This review summarizes bioactive compounds and its functions of (1) B. subtilis and its subspecies, (2) B. subtilis natto. Overall, natto is a natural source of bioactive compounds and is expected to be applied to functional foods. Natto, produced by fermentation of soybeans by B. subtilis natto (a subspecies of B. subtilis), has been consumed as a traditional fermented food for at least a thousand years. Natto is attracting interest as a potential functional food for health. This review summarizes compounds and its functions of B. subtilis natto.
Natto is a traditional fermented soybean-based food that has been an integral part of Japanese cuisine for several centuries. Although there have been extensive studies on the cognitive benefits of soybeans, only limited studies have examined the effects of natto on cognitive function. This study investigated the potential cognitive benefits of natto in senescence-accelerated mouse-prone 8 (SAMP8) mice. After 12 weeks of oral administering natto fermented for 18 h, the spatial learning and memory performance were improved compared with those in SAMP8 control mice. Furthermore, activation of the brain-derived neurotrophic factor (BDNF)/tropomyosin receptor kinase B (TrkB)/cAMP response element-binding protein (CREB) signaling and N-methyl-D-aspartate receptor (NMDAR)-calcium/calmodulin-dependent protein kinase II (CaMKII) cascade was observed in the hippocampus of SAMP8 mice that were fed natto. Additionally, natto administration upregulated trace amine-associated receptor 1 (TAAR1) as a modulator of NMDAR. These findings suggest that natto ameliorates cognitive decline by activating the TAAR1-mediated CaMKII/CREB/BDNF signaling pathway in the hippocampus of SAMP8 mice.
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The importance of plant secondary metabolites in food and their possible impacts on human health are the subjects of a steadily developing number of studies. Furthermore, as consumers become more conscious of diet-related issues, they desire natural ingredients that are anticipated to be safe and health-promoting; as a result, doum flour might be regarded as an excellent source of functional compounds. Hyphaene thebaica L. is a palm plant species with eatable ovoid fruits and a woody texture that is endemic to upper Egypt. It is one of the world's most significant and valuable plants. Doum flour is considered to contain higher moisture content, crude fiber, ash content, protein, fat, and vitamins, especially niacin, folic acid, pyridoxine, riboflavin, and thiamin. Previous findings showed that doum flour has high functional and nutritional qualities and may be used for a variety of key applications in the food sector. Doum nuts contain antioxidants as well as metabolites like tan-nins, saponins, steroids, glycosides, terpenes, and terpenoids. The results of different studies have indicated that doum flour is effective in various health maladies. This review highlights the anti-inflammatory, antioxidant, antibac-terial, anticancer, nutritional content, fertility activity, the rheological, and pharmacological ability of Hyphaene thebaica L. flour extracts and their significant phyto-constituents such as polyphenol, essential-oil, and flavonoid components. ARTICLE HISTORY
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Unlabelled: Nano-emulsions are receiving great attention in various industries, especially in the food sector. Peculiar properties of nano-sized droplets and high surface area are most suited for the development and delivery of functional ingredients. Nano-emulsions systems are suitable for encapsulation, protection, improving bioavailability, and target release of sensitive functional compounds. Nano-emulsions have promising potential for the delivery of nutraceuticals, probiotics, flavors, and colors. Nano-emulsions with active ingredients (antimicrobials) have a key part in ensuring food safety, nutrition, and quality of food. Nanoemulsions can also be used for biodegradable coating, packaging, antimicrobial coating, and quality and shelf life enhancement of different foods. The current review includes an overview of nanotechnology nano-emulsions, materials, techniques for formulation & production of nano-emulsions for food and nutrition. Furthermore, the analytical approaches used for the characterization of nano-emulsions and finally, the applications and limitations of nano-emulsions in the food industry are discussed in detail. Supplementary information: The online version contains supplementary material available at 10.1007/s13197-022-05387-3.
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Nanotechnology is currently a field of endeavor that has reached a maturation phase beyond the initial hypotheses with an undercurrent challenge to optimize the safety, and scalability for production and clinical trials. Lipid-based nanoparticles (LNP), namely solid lipid nanoparticles (SLN) and nanostructured lipid (NLC), carriers are presently among the most attractive and fast-growing areas of research. SLN and NLC are safe, biocompatible nanotechnology-enabled platforms with ubiquitous applications. This review presents a modern vision that starts with a brief description of characteristics, preparation strategies, and composition ingredients, benefits, and limitations. Next, a discussion of applications and functionalization approaches for the delivery of therapeutics via different routes of delivery. Additionally, the review presents a concise perspective into limitations and future advances. A brief recap on the prospects of molecular dynamics simulations in better understanding NP bio-interface interactions is provided. Finally, the alliance between 3D printing and nanomaterials is presented here as well.
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Celiac disease is a common immune-mediated disease characterized by abnormal T-cell responses to gluten. For many patients, symptoms and intestinal damage can be controlled by a gluten-free diet, but, for some, this approach is not enough, and celiac disease progresses, with serious medical consequences. Multiple therapies are now under development, increasing the need for biomarkers that allow identification of specific patient populations and monitoring of therapeutic activity and durability. The advantage of identifying biomarkers in celiac disease is that the underlying pathways driving disease are well characterized and the histological, cellular, and serological changes with gluten response have been defined in gluten challenge studies. However, there is room for improvement. Biomarkers that measure histological changes require duodenal biopsies and are invasive. Less invasive peripheral blood cell and cytokine biomarkers are transient and dependent upon gluten challenge. Here, we discuss established biomarkers and new approaches for biomarkers that may overcome current limitations.
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Bacillus subtilis is widely used to produce a variety of fermented soybean foods in Asian countries. These foods are classified into two types: salted (e.g., tuong of Vietnam) and unsalted (e.g., natto of Japan). To explore the effect of salt on the fermentation process, cell growth and the extracellular production of poly-γ-glutamic acid (γPGA), which confers the sticky texture to natto, were examined under high salt conditions (0.5-1.5 M NaCl). The expression of the γPGA synthetic gene was monitored using the lacZ-fusion reporter method, and the production of γPGA was detected by immunoelectrophoresis. Although B. subtilis (natto) does not usually produce γPGA in carbon source-limiting media such as LB broth, it was found to synthesize γPGA when excess salt was added to the medium. The need for genes such as degU, degQ, comP and sigB for this salt-induced γPGA production suggests that both the general stress response induced by sigma B and activation of DegU protein through phosphorylation are involved in this process. © 2018, Japan International Research Center for Agricultural Sciences.
The fast progress in nanomedicine and nanoparticles (NP) materials presents unconventional solutions which are expected to revolutionize health care with great potentials including, enhanced efficacy, bioavailability, drug targeting, and safety. This review provides a comprehensive update on widely used organic and inorganic NP with emphasis on recent development, challenges and future prospective for bio applications where, further investigations into innovative synthesis methodologies, properties and applications of NP would possibly reveal new improved biomedical relevance. NP exhibit exceptional physical and chemical properties due to their high surface area to volume ratio and nanoscale size, that led to major breakthroughs in therapeutic, diagnostic and screening techniques where innovative design engineering allowed a paradigmatic shift in their market share. Finally, an update of FDA approved NP is explored where innovative design engineering allowed a paradigmatic shift in their market share. This review would serve as a discerning comprehensive source of information for learners who are seeking a cutting-edge review but have been astounded by the size of publications.
Melanoma is one of the most dangerous malignant epidermal cancers. Natto freeze-drying extract (NFDE) and natto water extract (NWE) were isolated from natto, soybeans fermented by Bacillus subtilis natto, which were assessed as potential anti-melanoma agents. Cell cytotoxicity assays revealed significant anti-melanoma effects of NFDE and NWE in a dose-dependent manner, and exhibited low influences on normal skin cells, including Hs68, HaCaT and adipose tissue-derived stem cells (ADSCs), respectively. Through a flow cytometer assay and autophagy acridine orange staining, the cellular death phenomenon shifted from autophagy to apoptosis with the increased dosages. Reactive oxygen species (ROS) were enhanced using DCFDA (2,7-dichlorodihydrofluorescein diacetate) staining when melanoma cells were treated with the extract. NFDE and NWE treatments increase the oxidative stress of cancer cells and cause apoptosis by inhibiting AMP-activated protein kinase (AMPK). NFDE and NWE were considered to play a critical role in cell death through ROS adjustment, autophagy regulation and apoptosis promotion.
Millet bran was fermented with Bacillus natto and the changes of structural, physicochemical and functional properties of its dietary fiber were investigated. Results showed that B. natto fermentation enhanced soluble DF content from 2.3% to 13.2%, and soluble DF/insoluble DF ratio from 3.1% to 19.9%. SEM and FTIR assay indicated that fermentation led to the degradation of cellulose and hemicellulose, thereby forming more porous and loose structure and polysaccharides. The binding capacities such as water and oil holding capacity, swelling capacity as well as cholesterol, bile salts, nitrite ion and glucose adsorption capacity were improved, while cation exchange capacity was not significantly changed. The total phenolic content and DPPH free radical scavenging capacity increased significantly. Overall, fermentation of millet bran by B. natto improved the structural and functional properties of its DF, which could be applied as a functional ingredient in food products.
Fermented soybean foods made with Bacillus subtilis cells are produced in China, and they are called dou chi (or dauchi). They include salted, sweet, and nonsalted types. The salted type of dou chi (xian-dou chi) contains 10% to 20% salt to inhibit their putrefaction by contaminating bacteria. The most typical sweet dou chi is called tian-dou chi: it is used as a seasoning for Beijing duck. Nonsalted dou chi has been developed into various kinds of natto.¹ (See Chapter 16 for more information on Chinese fermented foods.) Food of this type is called itohiki-natto (hereafter shortened to “natto”) in Japan, kinema in Nepal and Myanmar, tua nao in Thailand, and chungkuk-jang in Korea. Natto is produced only with B. subtilis (natto) (formerly called B. natto, see Section 2.1).
Human-derived dipeptidyl peptidase IV (DPPIV) is a costly material used in the discovery of drugs for diabetes. In this study, we demonstrated the efficacy of DPPIV from Aspergillus oryzae as an alternative to human-derived DPPIV for identifying DPPIV inhibitors. Fermented soybean, also called natto, suppresses blood glucose levels in humans; however, the underlying mechanism remains unknown. This study determined the in vitro DPPIV inhibitory activity of isolated peptides from natto. Amino acid sequences of two peptides isolated from natto were identified by LC/MS/MS as Lys-Leu and Leu-Arg. These isolated peptides inhibited DPPIV in a dose-dependent manner, with IC50 values of 41.40 ± 2.68 and 598.02 ± 18.35 μg/ml, respectively. These results indicate the potential mechanism of blood glucose control by natto and novel roles of Lys-Leu and Leu-Arg as DPPIV inhibitors.