B2B Proposal

Development of Gluten-Degrading Probiotics

Increased Demand for Wheat

Flour foods, such as bread, pasta, and noodles, are popular and loved by almost everyone. Flour, which is chewy, sticky, and crunchy depending on the recipe, is used almost indiscriminately in modern food. Flour is one of the most important raw ingredients in the Korean food industry and its main uses are for making noodles, confectionery, and baking. According to Korea Flour Mills Industrial Association (KOFMIA), the total flour production in 2020 amounted to about 174 million tons.

Flour produced in Korea is divided into strong flour, all-purpose (medium) flour and cake (soft) flour, and there are mixed flour and whole-wheat flour being produced since 1986. While mixed flour belongs to the all-purpose flour for noodle making, whole-wheat flour is pulverized wheat that is used for baking and confectionery purposes. 74.5% of the total flour production in 2020 was all-purpose flour, while 16.2% was strong flour and 9.3% was cake flour.

Strong flour is mainly made from American Dark Northern Spring (DNS) wheat, and Canada Western Red Spring (CWRS) wheat has also been used for baking since 1991. All-purpose flour is produced by mixing Western White wheat and Hard Red Winter wheat in equal proportions (when using American wheat as a raw ingredient), and it is mainly used for making noodles and household multipurpose flour.

In the case of flour, hundreds of products optimized for each food have been produced through systematic research accumulation and industrialization for over 100 years. In Western societies where wheat is the staple food, most wheat-processed foods have developed into bread, cookies, noodles and pizza. In the case of Korea, food culture is increasingly becoming westernized, and as a result, processed products using wheat are diversifying.

Although rice is Korea’s staple food, annual per capita rice consumption has decreased from 99.2kg in 1998 to 74.0kg in 2009. The annual per capita rice consumption in Korea declined to 59.2kg by 2019, while consumption of wheat-processed foods is steadily increasing (Statistics Korea, 2020). The total consumption of processed flour in Korea has been steadily increasing since 1965, reaching a total of 2 million tons in 2019, showing a continuous increase (KOFMIA, 2020).

However, grains such as wheat and barley contain gluten (an insoluble protein) along with starch, which can cause digestive disorders, gluten intolerance, gluten allergy, and inflammatory bowel disease when consumed. According to a survey by the World Allergy Organization, the ranking of allergen-causing foods in Korea is milk > egg > wheat > walnut, almond > peanut. Although wheat accounts for a large proportion of allergens, public awareness of gluten in Korea is currently very low. As the consumption of wheat continues to increase, there is concern that the population suffering from various conditions caused by wheat and gluten will also increase.

Gluten is an insoluble protein complex present in grains such as wheat and barley. Its viscosity and elasticity make it easy to form dough, and it serves to collect the carbon dioxide from the fermented dough so that it does not escape and inflate the bread or cake. Gluten protein is largely composed of two proteins – glutenin (responsible for the elasticity of dough) and gliadin (responsible for the extensibility and viscosity of dough). When wheat flour is mixed with water, gliadin and glutenin combine to form gluten in a web-like structure, and the ratio of these two proteins varies depending on the type of flour.

Composition of gluten protein

On average, strong flour contains more than 12% total protein, all-purpose (medium) flour contains 9-12%, and cake (soft) flour contains less than 9% total protein. In the case of strong flour with the highest protein content, the most gluten is formed during kneading which results in a chewy and elastic dough. Thus, strong flour is mainly used for making bread due to its well-maintained pores and good swelling properties. On the other hand, cake flour, which is relatively soft with gluten and produces a soft or fluffy texture without stickiness, is mainly used in confectionery products such as cakes, cookies, and scones. As such, the ratio of gluten in wheat flour affects not only the volume, color and texture of the dough but also affects various aspects such as the formation of the dough skeleton, the gas retention capacity generated during fermentation, and the presence or absence of pores and swelling of the dough.

By washing off the starch in the wheat dough,
a ball of pure yellowish-beige gluten protein is earned, which can be stretched into a thin film

What is Gluten?

Health Problems Caused by Wheat Gluten

Gluten has excellent water absorption properties due to its structure, and thus when it enters the stomach, it interferes with the digestive juices that help digestion from functioning properly. As a result, it causes various digestive disorders in the gastrointestinal tract. In particular, of the two proteins that make up gluten, glutenin degrades whereas gliadin moves to the large intestine without being broken down properly in the stomach and small intestine. Instead of excreting through feces, most gliadin properties stick to the mucous membrane of the large intestine and accumulate, which leads to decay due to accumulation of stool. Moreover, these gliadin properties that stay in the intestine and generate harmful toxins, which can cause intestinal diseases. Also, the beneficial bacteria in the intestines decrease and the harmful bacteria increase, causing the normal intestinal microbial balance of 85:15 to be broken, a state that is called, “dysbiosis.”

In other words, when gluten enters our body through our favorite wheat flour foods, it can cause the following health problems without us even being aware of it:

(1) Gluten Intolerance
Gluten intolerance is simply a kind of rejection reaction caused by our body's inability to break down gluten. Its symptoms include diarrhea, abdominal pain, constipation, fatty stool (fat appears in feces), bloating, and abdominal distension. Also, mood swings, migraine headaches, fatigue, and anxiety may be present as symptoms accompanying digestive disorders.

(2) Gluten Allergy
The immune system recognizes gluten as a foreign substance and triggers an overreaction, which leads to symptoms of hyperimmune reaction such as atopic dermatitis, skin rash, facial edema, muscle pain, cough and asthma, and allergic rhinitis (sneezing, runny nose, stuffy nose). If the reaction is severe, it may lead to anaphylaxis (hypotensive shock), in which the pulse rises and blood pressure drops sharply following respiratory difficulty due to airway edema.

(3) Gluten Irritable Bowel Disease
To those who are extremely sensitive to gluten, gluten may trigger an inflammatory response in the immune system, which causes atrophy or loss of the villi in the intestinal mucosa, making it impossible to absorb nutrients. This then leads to health problems such as abdominal pain, diarrhea, malabsorption, weight loss, malnutrition, and bloating. As a result, those who have been diagnosed with gluten irritable bowel disease must follow a gluten-free diet for the rest of their lives.

Gluten-induced damage to the villi of the small intestine can lead to gluten irritable bowel disease.

Leaky Gut Syndrome (LGS)
The intestinal mucosal cells, which act as the first line of defense in our body's immune system, are composed of a single cell layer and maintain a certain gap between cells, called tight junctions. This close bond is maintained by zonulin, and when gliadin in gluten promotes the secretion of zonulin and zonulin is oversecreted, the close bond between the villi opens and increases the permeability of the intestinal mucosa, resulting in “leaky gut syndrome.” With increased permeability due to dismantled gaps, foreign substances with large molecular weight that have not been decomposed, viruses, bacteria, antibiotics, heavy metals, and residual pesticides can enter the blood vessels and contaminate the blood. This results in immune system diseases, such as atopic dermatitis, acne, seborrheic dermatitis, psoriasis, eczema, and urticaria.

Various toxins and harmful substances enter through the dismantled tight junction gaps, causing excessive immune response and inflammation

Health Problems Caused by Excessive Consumption of Carbohydrates

The structure of wheat, which is the main raw material for wheat flour, can be largely divided into the bran, aleurone layer, endosperm, and germ. The aleurone layer, just below the outer bran layers, is rich in minerals, vitamins, proteins, antioxidants, and fat, whereas the endosperm contains mostly only starch. When flour is milled, the bran and germ are removed from the endosperm, and the endosperm is then refined and ground into flour particles. Therefore, the intake of refined flour means the intake of high carbohydrates excluding nutrients, which causes nutritional imbalance and excessive carbohydrate intake. Refined simple sugar quickly raises blood sugar and promotes insulin secretion when ingested, which in turn can cause health problems such as obesity, diabetes, non-alcoholic fatty liver, and ateriosclerosis.

Structure of Wheat and Gluten

Excessive Craving for Wheat Flour
Gluten accumulated in the body through wheat produces an amino acid complex called “exorphins,” which stimulates the central nervous system. Exorphins are known to act on the same receptors that morphines act on in the human brain, and thus, eating wheat flour can cause a feeling of joy, happiness, and satisfaction as if endorphins were secreted. This can cause “flour intoxication” by making people want to eat wheat flour over and over again, and excessive consumption of wheat flour products increases the risk of developing blood acidification, obesity, diabetes, arteriosclerosis, and fatty liver.

Is gluten-free healthy food?

Gluten-free foods are foods commercialized to alleviate health problems caused by gluten. In order to eat gluten-free, people must commit to a lifetime restriction on gluten intake and avoiding grains such as wheat, rye, barley and oats. However, strictly restricting gluten is not an easy task. Gluten is a protein also used to thicken products or give food elasticity and a sponge-like feel, and it is commonly found in sauces, chocolate, jam, ice cream, and fish cakes and sausages.

As the number of patients with chronic digestive disorders showing gluten sensitivity increases, bread and confectionery excluding gluten ingredients (referred to as “gluten-free” foods) are on the rise, and their presence is expanding as a kind of functional food. According to a consumer study conducted by Packaged Facts, 18% of adult consumers consume gluten-free foods, and consumer perceptions that gluten-free foods are good for their health are driving their purchase. Also, as awareness of food allergy diseases caused by wheat or other food allergens is increasing recently, consumers' interest in gluten-free products is continuously rising.

The consumption of wheat instead of rice is increasing due to the westernization of dietary life, and more and more people are complaining about health problems, such as digestive disorders, caused by gluten. As a result, interest in gluten-free products is increasing. The preference for gluten-free products is also growing in Korea following this market trend and various domestic gluten-free products are rushing to market.

However, conventional gluten-free foods are manufactured using gluten-free or low-content grains (rice, sorghum, quinoa, crude oats, buckwheat, etc.) to replace wheat starch so that the total gluten content is 20ppm or less, or manufactured by completely replacing protein (gluten), which makes up about 10% of wheat flour, with carbohydrates such as potato or corn starch. This eventually achieves close to 100% carbohydrate content. As a high-carbohydrate diet, a gluten-free diet can ultimately lead to a lack of nutrients such as iron, calcium, thiamine, riboflavin, niacin, and folic acid. Therefore, more and more research studies are being published that emphasize the importance of considering one’s health and nutritional status before committing to a gluten-free diet as it can otherwise lead to adverse health effects.

Gluten-Degrading Probiotic

Microbiome Inc. has developed a unique lactic acid bacteria product to help break down gluten in wheat and enhance digestion, and to alleviate the health problems caused by gluten, such as “gluten intolerance,” “gluten allergy,” and “gluten-sensitive bowel disease.”

Microbiome Inc.’s Gluten-Degrading Probiotic was developed based on De-glu MB0601 and De-glu GLU70 strains, which were isolated from fermented foods and have demonstrated excellent gluten-degrading ability among various lactic acid bacteria strains. De-glu MB0601 received a joint patent for its gluten-degrading ability through joint research and development by Microbiome Inc. and a government-funded research institute. De-glu GLU70 was also recognized for its gluten-degrading ability, and Microbiome Inc. received a patent for its effectiveness in preventing and improving inflammatory bowel disease caused by gliadin.

Upon applying De-glu MB0601 and De-glu GLU70 strains to gluten-containing grains (wheat, rye, barley, oats, etc.) and fermenting them, the results showed that the gluten present in the grains were degraded.

Moreover, in animal model experiments, De-glu MB0601 increased secretion of gastrin hormone, which helps promote gastric juice secretion, and it also increased secretion of pancreatic amylase, an enzyme that breaks down carbohydrates in wheat. De-glu GLU70 was effective in inhibiting intestinal epithelial cell damage as well as improving inflammation caused by gliadin in gluten. By using the gluten-degraded flour produced through fermentation as described above, it is possible to manufacture various foods containing wheat as a main ingredient (such as ramen, noodles, udon, pizza, and bread) without worrying about potential negative health effects caused by gluten. Therefore, Microbiome Inc.’s gluten-degrading probiotic is expected to be applicable to various processed food fields where gluten-free is popularly applied: such as confectionery, cereals, sauces, desserts, energy bars, meat/meat substitutes, snacks, and many more.

Microbiome Inc.’s Gluten-Degrading Probiotic includes 4 different beneficial bacteria:

De-glu MB0601 (gluten-degrading ability, enhances digestion)

De-glu GLU70 (gluten-degrading ability, prevents and improves intestinal

inflammation)

5-Strain Probiotic Complex with confirmed antibacterial and antiviral effects

Digestive Enzyme-Producing Strain that helps break down carbohydrates and

protein

In addition, there are also 7 types of postbiotics, or heat-treated, dried cultures of beneficial lactic acid bacteria:

De-glu MB1 (heat-treated De-glu MB0601 strain)

De-glu MB2 (heat-treated De-glu GLU70 strain

Dipro-rhamnosus heat-treated strain (made using Dipro-rhamnosus strain

effective in preventing and treating obesity)

MB201, MB202, MB203, and MB204 which are heat-treated lactic

acid bacteria; contain 17 types of amino acids, and various organic acids as
well as short-chain fatty acids according to a nationally recognized test analysis.

In total, 11 types of lactic acid bacteria components make up Microbiome Inc.’s Gluten-Degrading Probiotic.

Unlike conventional gluten-free foods, the Gluten-Degrading Probiotic helps prevent and improve indigestion and health problems caused by gluten without modifying the unique nutritional components contained in wheat. Moreover, it can be easily sprinkled on wheat flour foods or consumed with water at any time, and can also be used in various ways, such as kneading directly into flour dough during cooking. Through modern microbiome technology, Microbiome Inc. has secured a safe ingredient that will enable people to enjoy flour food with confidence and worry-free.

For Restaurants/Bakeries (1kg)

Home Use (Large, 100g)

Home Use (Small, 2.5gx20)

Travel Size (2.5gx5)

Microbiome Inc. Gluten-Degrading Probiotics

Gluten-Degrading Probiotic: Lactic acid bacteria

By conducting a joint research and development with a government-funded research institute, Microbiome Inc. was able to develop De-glu MB0601 through isolation from domestic fermented foods. Our lab results confirmed that fermenting wheat flour dough with MB0601 strain led to approximately 70% gluten degradation. De-glu MB0601 was officially patented as “MB0601 strain with gluten-degrading ability.”

Electron microscope image of De-glu MB0601

De-glu MB0601

Gluten Developed

Gluten degraded

We also conducted a lab experiment in which mice were fed meals that were mixed with our gluten-degrading flour (fermented with with De-glu MB0601) or ordinary flour for 8 weeks. The results showed that the blood gastrin content of the mice increased by up to 60% and the secretion of pancreatic amylase increased by up to 80% as the content of gluten-degrading flour increased. Gastrin (a hormone secreted by G cells of the gastric glands) promotes gastric juice secretion and aids digestion, and pancreatic amylase is an enzyme that breaks down carbohydrates in wheat flour. Therefore, the results revealed that consuming food made using the gluten-degrading flour provides better digestion compared to food made using ordinary flour.

Blood gastrin levels according to feeding gluten-degraded flour

Pancreatic amylase levels according to feeding gluten-degraded flour

De-glu GLU70

The official patent for Microbiome Inc.’s De-glu GLU70 acknowledges De-glu GLU70’s gluten-degrading ability and that it is effective in “prevention, improvement or treatment of inflammatory bowel disease caused by gliadin.”

Electron microscope image of De-glu GLU70 (top)

Colonies grown on agar plate (bottom)

Certificate of Patent for De-glu GLU70

In addition to helping break down gluten, it has been confirmed that De-glu GLU70 also inhibits intestinal inflammation and damage to intestinal epithelial cells caused by the gliadin in gluten. Upon conducting an experiment in which De-glu GLU70 was mixed with food containing gliadin and fed to mice for 21 days, the results showed that De-glu GLU70 inhibited damage to the intestinal mucosa caused by gliadin, as the villi and the crypt located in the intestinal tissue of the normal control group (NC) showed no abnormalities. However, the group of mice that were fed the general diet containing gliadin (without De-glu GLU70) displayed significant increase in the inflammatory cell infiltration (black arrows in the figure below) in the villi and severe intestinal epithelial cell degeneration and basal edema. Moreover, moderate intestinal epithelial cell degeneration and basal edema were observed in the group ingested at a low concentration of De-glu GLU70, while only weak intestinal epithelial cell degeneration was observed in the group ingested at a high concentration of De-glu GLU70. As no other clinical findings were observed, it was confirmed that the De-glu GLU70 inhibited intestinal tissue damage caused by gliadin.
Also, the secretion of the inflammatory cytokine TNF-alpha caused by gliadin decreased, and the secretion of the anti-inflammatory cytokine IL-10 increased in the group ingested with De-glu GLU70. Also, the protein expression of inflammatory cytokines decreased compared to the group that did not ingest De-glu GLU70, which showed that De-glu GLU70 can suppress the inflammation induced by gluten.

Confirmed inhibition of gluten(gliadin)-induced degeneration of intestinal epithelial cells by GLU70

Normal intestinal

epithelial cell

Intestinal epithelial cells
damaged by gliadin

Degeneration of damaged intestinal epithelial cells inhibited by De-glu GLU70

Decreased secretion of TNF-α (inflammatory cytokine)

Increased secretion of IL-10
(anti-inflammatory cytokine)

Decreased expression levels of inflammatory
proteins in De-glu GLU70-ingested groups

5-Strain Probiotic Complex

The 5-strain lactic acid bacteria complex included in the gluten-degrading lactic acid bacteria were isolated from domestic fermented foods. A nationally recognized test analysis confirmed that they have antibacterial and antiviral effects, and help to maintain the balance of intestinal microbes, regulate intestinal pH and inhibit the proliferation and activity of harmful intestinal bacteria and viruses.

Digestive Enzyme-Producing Strain

The ‘digestive enzyme-producing strain’ contained in our gluten-degrading lactic acid bacteria produces various enzymes that break down carbohydrates including starch as well as proteins and promotes the secretion of amylase to facilitate the breakdown, absorption, and digestion of flour. Therefore, it also helps to improve digestive problems caused by bloating or gas when eating wheat flour foods.

De-glu MB1 (Heat-treated culture of De-glu MB0601 strain)

De-glu MB1 is a heat-treated, dried culture of the patented strain ‘De-glu MB0601,’ which has gluten-degrading ability. It is a heat-treated, dried culture with the unique gluten-degrading ability of the De-glu MB0601 strain as well as a function to help the digestion of flour such as promoting gastrin secretion and pancreatic amylase secretion. De-glu MB1 also contains beneficial metabolites, such as organic acids and short-chain fatty acids, which help to create a healthy intestinal environment by increasing beneficial intestinal bacteria, inhibiting the growth of harmful bacteria, and maintaining the ideal intestinal microbiome balance of 85:15.

De-glu MB2 (Heat-treated culture of De-glu GLU70 strain)

De-glu MB2 is a heat-treated, dried culture of the patented strain ‘De-glu GLU70,’ with gluten-degrading ability. It is a postbiotic that Microbiome Inc. received patent for its effectiveness in “preventing and improving inflammatory bowel disease caused by gliadin.”

As a result of feeding the heat-treated, dry culture of De-glu GLU70 strain to mice for 21 days, it was found that the degeneration and edema of intestinal epithelial cells caused by gliadin were suppressed in the group that consumed the postbiotic compared to the group that did not.

Moreover, the secretion of the inflammatory cytokine TNF-alpha induced by gliadin decreased and the secretion of the anti-inflammatory cytokine IL-10 increased in the group that ingested the heat-treated dry culture of De-glu GLU70. It was also discovered that the protein expression of inflammatory cytokines decreased compared to the group that did not ingest the heat-treated dry culture of De-glu GLU70. Therefore, it was confirmed that the heat-treated dry culture of De-glu GLU70 can suppress intestinal inflammation caused by gliadin in gluten.

Confirmed inhibition of gluten(gliadin)-induced degeneration of intestinal epithelial cells by De-glu MB2

Normal intestinal

epithelial cell

Intestinal epithelial cells
damaged by gliadin

Degeneration of damaged intestinal epithelial cells inhibited by De-glu MB2

Reduced secretion of inflammatory cytokines, increased secretion of anti-inflammatory cytokines,
and decreased expression of inflammatory proteins when feeding heat-treated, dry De-glu GLU70
strain with feed (containing gliadin)

Dipro-rhamnosus heat-treated strain

Dipro-rhamnosus heat-treated dry culture contained in our gluten-degrading probiotics is a heat-treated, dry culture product of the patented Dipro-rhamnosus strain. This lactobacillus strain was patented for its “preventive or therapeutic effect on obesity.”

As a result of animal model experiments using mouse precursor adipocytes in patent clinical research, it was confirmed that brown fat was induced by treatment with rhamnosus strain, promoting “browning” of the adipocytes. (Browning: white fat cells that store energy consume energy and change into brown fat cells that maintain heat homeostasis and exothermic reaction) It was also confirmed that the gene expression of mouse beige adipocytes and brown adipocytes was significantly increased compared to the control group not treated with the strain.

Also, upon administering Dipro-rhamnosus strain or the culture medium for 12 weeks to obese mice induced by a high-fat diet, they showed a weight gain inhibition effect of 5.7% compared to the negative control.

Results of 12 weeks of administration of rhamnosus strain or culture medium to obese mice induced by a high-fat diet: confirmed the effect of inhibiting weight gain by 5.7% compared to the negative control

Heat-treated lactic acid bacteria MB201, MB202, MB203, MB204

Heat-treated, dried lactobacillus cultures MB201, MB202, MB203, MB204 contained in the gluten-degrading probiotics are heat-treated, dried products of 4 strains of lactic acid bacteria isolated from fermented foods. In addition to the beneficial functions of these lactic acid bacteria themselves, there are 17 kinds of amino acids and various organic acids and short-chain fatty acids (confirmed by a nationally recognized test analysis) that are beneficial to the intestinal health.

Analysis result for organic acids and short chain fatty acid contents of MB201~204 (Left)
Analysis result for amino acid content of MB201~204 (Right)

Microbiome Inc.‘s four lactic acid bacteria strains for MB201, MB202, MB203, MB204 have been deposited and accepted by Korean Collection for Type Cultures, an international depositary authority

Evaluation of gluten degradability of De-glu MB0601 strain

Microbiome Inc.‘s five lactic acid bacteria strains have been deposited and accepted by Korean Collection for Type Cultures, an international depositary authority