KR20240055598A - Novel strain of Bacillus velezensis and uses thereof - Google Patents

Abstract

The present invention relates to a new Bacillus belegensis strain,
The Bacillus velezensis GV1 strain deposited under the deposit number KCTC 15222BP of the present invention can exhibit acid resistance, bile resistance, and gluten-decomposing activity, and can be widely used in food, pharmaceutical, and feed compositions.

Description

Novel strain of Bacillus velezensis and uses thereof}

The present invention relates to a new Bacillus belegensis strain and its use for acid resistance, bile resistance, and gluten decomposition.

Probiotics have generally been defined as live microbial food additives with health benefits (Lilly and Stillwell, Science 147:747-748, 1965), but more recently, Salminen et al. (Int. J. Food Microbiol. 44:93-106, 1998) ) is defined as a live microbial agent contained in food, feed, or dietary additives designed to improve the health of humans or animals. The bacteria most commonly used as probiotics are lactic acid bacteria, including Lactobacillus and Streptococcus.

Dead lactic acid bacteria have various differences compared to live bacteria. Dead cells do not settle in the intestines and become active like live bacteria, but contribute to immune regulation through direct contact with immune cells. Since they are not live bacteria, they are not affected by harsh environments (physical and chemical digestion processes) in the gastrointestinal tract. It is heat stable, maintains consistent functionality during storage and delivery, and can be applied to a variety of products that have undergone a sterilization process. Thanks to these characteristics of material stability, not only live bacteria but also dead bacteria are applied to health functional foods, general foods, medicines, animal feed, and cosmetic raw materials (US Patent No. 8361481). In Japan, the market for dead lactic acid bacteria is the entire lactic acid bacteria market. Dead lactic acid bacteria, or parabiotics, account for more than 25% of the total. Only the genera Lactobacillus, Bifidobacterium, and Enterococcus are used as dead cells. Bacillus forms endospores, so it is very difficult to kill bacteria, and cross-contamination with other bacteria occurs. Due to this problem, the industry is reluctant to cultivate Bacillus strains and kill them, making commercialization difficult.

Against this background, the present inventors made intensive research efforts to develop an agent that can safely and effectively enhance immunity. As a result, they confirmed that a new Bacillus belegensis strain can effectively enhance immunity, and completed the present invention.

One object of the present invention is to provide a Bacillus velezensis GV1 strain deposited with the deposit number KCTC 15222BP.

Another object of the present invention is to provide a food composition containing the above strain.

Another object of the present invention is to provide a pharmaceutical composition containing the above strain.

Another object of the present invention is to provide a feed composition containing the above strain.

Another object of the present invention is to heat treat the Bacillus velezensis GV1 strain deposited under the accession number KCTC 15222BP at 100°C to 150°C for 5 to 30 minutes to kill the strain. To provide a method for producing dead cells.

This is explained in detail as follows. Meanwhile, each description and embodiment disclosed in the present invention may also be applied to each other description and embodiment. That is, all combinations of the various elements disclosed in the present invention fall within the scope of the present invention. Additionally, the scope of the present invention cannot be considered limited by the specific description described below.

Additionally, those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Additionally, such equivalents are intended to be encompassed by this invention.

One aspect of the present invention provides a Bacillus velezensis GV1 strain deposited with deposit number KCTC 15222BP.

In the present invention, the term “Bacillus genus” refers collectively to rod-shaped aerobic or facultative anaerobic Gram-positive bacteria widely distributed in nature, also known as bacilli. Bacillus strains commonly found in nature include free-living types and pathogenic types. It is known that under stressful environmental conditions, egg-shaped endospores can be produced and remain dormant for extended periods.

As used herein, the term “strain” refers to a collection of individuals derived from a single cell, proliferated through asexual reproduction, and having identical genetic characteristics. Within a species, genetic characteristics are not present. Several different genetic variants exist. In the present invention, the strain refers to Bacillus belegensis, and may specifically be Bacillus belegensis GV1 deposited under the accession number KCTC 15222BP, but is not limited thereto.

Bacillus velegensis GV1 of the present invention has a 16s rDNA base sequence represented by SEQ ID NO: 1. The Bacillus belegensis of the present invention may be derived from ginseng vinegar, but is not limited thereto.

The strain of the present invention has i) acid resistance; ii) endobiliary; and iii) gluten decomposition activity.

The term "acid resistance" in the present invention refers to the property of withstanding high acidity. If the strain has acid resistance, it can be prevented from being decomposed or damaged even if it is ingested through various administration routes, including oral, and exposed to strong acidic conditions in the stomach.

In one embodiment of the present invention, it was confirmed that the Bacillus velegensis GV1 strain maintains a very high survival rate (growth rate) in pH 2 to 6, specifically pH 2 to 4, which is a strong acid condition, and more specifically, pH 2 in a stomach acid environment.

As used herein, the term “biliary resistance” refers to resistance to digestive enzymes in bile. Bile is a slightly alkaline greenish-brown liquid that is produced in the liver and stored in the gallbladder. It helps digest fat in the glandular intestine inside the small intestine. It emulsifies fat and aids digestion and absorption. This bile acts on strains ingested through various routes, including orally, and is one of the main causes of reducing the effectiveness of strain administration.

In one embodiment of the present invention, the Bacillus velegensis GV1 strain has bile tolerance to bile salts similar to the intestinal environment, and contains 0.1 to 1.5% (w/v) bile salts, specifically 0.15% to 1% (w). /v) bile salts, more specifically, it was confirmed that excellent tolerance was maintained even at 0.3% to 0.5% (w/v) of bile salts.

It can be seen that Bacillus velegensis GV1 of the present invention has excellent acid resistance and bile resistance and has a high survival rate in the environment leading to the stomach and intestines when ingested.

The term "gluten" of the present invention is an insoluble protein contained in grains such as wheat and barley, and is a mixture of several proteins. It is an immunoreactive peptide that is difficult to digest in the gastrointestinal tract (typically a 26-mer peptide derived from γ-gliadin, There is a 33-mer peptide derived from α-gliadin, which causes various gastrointestinal disorders and at the same time stimulates a negative immune response (T cell activation), thereby inducing an inflammatory response and causing various symptoms (fatigue, skin trouble, reflux esophagitis, irritable bowel syndrome). etc.). In the present invention, “gluten decomposition” may mean decomposing the non-degradable gluten of wheat flour to prevent the gluten or gliadin from being toxic, thereby preventing gastrointestinal disorders or facilitating digestion.

In the present invention, the Bacillus velegensis GV1 strain may exhibit proteolytic activity, and may specifically hydrolyze the recalcitrant gluten of wheat flour, but is not limited thereto.

In one embodiment of the present invention, the Bacillus velegensis GV1 strain was inoculated into a medium containing gluten at a concentration of 1% and the change in turbidity was observed with culturing time. As a result, it was confirmed that the turbidity decreased.

Through this, the Bacillus velegensis GV1 strain of the present invention is a safe lactic acid bacterium that can be used in food, and is expected to be able to produce gluten-free processed foods by fermenting wheat flour. It can be used to prevent cancer and gluten allergies.

The strain of the present invention may be a dead cell, and specifically, the dead cell may be prepared by heat-treating Bacillus belegensis, and more specifically, the Bacillus belegensis GV1 strain deposited under the deposit number KCTC 15222BP or the above strain. It may be prepared by heat treating the culture at 100°C to 150°C for 5 to 30 minutes.

In the present invention, the term "dead cell" refers to a form produced through heat treatment of live cells in which the strain is cultured, and is a form in which the activity of the strain is prevented from occurring, including cytoplasm, antibacterial active substances such as bacteriocin, organic acids, etc. may include. In the present invention, the dead cells are Bacillus velegensis GV1 strain deposited under the deposit number KCTC 15222BP or a culture of the strain at 100°C to 150°C, specifically 105°C to 140°C, more specifically 110°C to 130°C for 5 days. It may be manufactured through heat treatment for minutes to 30 minutes, specifically 7 minutes to 25 minutes, and more specifically 10 minutes to 20 minutes.

“Culture” of the present invention refers to a product obtained by culturing the above strain in a medium. For example, the culture of the present invention may contain components remaining in the culture medium of Bacillus belegensis after harvesting the strain.

The culture may be the entire culture of the Bacillus belegensis strain, culture supernatant, lysate, fractions thereof, etc. At this time, the culture supernatant can be obtained by centrifuging the culture of the strain, the lysate can be obtained by physically or sonicating the strain, and the fraction is the culture, culture supernatant, lysate, etc. It can be obtained by applying methods such as centrifugation and chromatography.

The medium and other culture conditions used for cultivating the strain of the present invention can be any medium used for cultivating ordinary Bacillus genus microorganisms without particular limitation. Specifically, the strain of the present invention can be cultured with an appropriate carbon source, nitrogen source, and human resources. , can be cultured under aerobic or anaerobic conditions in a conventional medium containing inorganic compounds, amino acids, and/or vitamins, while controlling temperature, pH, etc.

In the present invention, the carbon source includes carbohydrates such as glucose, fructose, sucrose, maltose, etc.; Sugar alcohols such as mannitol, sorbitol, etc., organic acids such as pyruvic acid, lactic acid, citric acid, etc.; Amino acids such as glutamate, methionine, lysine, etc. may be included, but are not limited thereto. Additionally, natural organic nutrient sources such as starch hydrolyzate, molasses, blackstrap molasses, rice bran, cassava, bagasse and steep liquor can be used, as well as glucose and pre-sterilized molasses (i.e. molasses converted to reducing sugars). The same carbohydrates can be used, and various other carbon sources of appropriate amounts can be used without limitation. These carbon sources may be used individually or two or more types may be used in combination.

The nitrogen source includes inorganic nitrogen sources such as ammonia, ammonium sulfate, ammonium chloride, ammonium acetate, ammonium phosphate, anmonium carbonate, and ammonium nitrate; Organic nitrogen sources such as amino acids, peptone, NZ-amine, meat extract, yeast extract, malt extract, steep liquor, casein hydrolyzate, fish or its decomposition products, defatted soybean cake or its decomposition products, etc. can be used. These nitrogen sources may be used alone or in combination of two or more types, but are not limited thereto.

The agent may include monopotassium phosphate, dipotassium phosphate, or a corresponding sodium-containing salt. Inorganic compounds include sodium chloride, calcium chloride, iron chloride, magnesium sulfate, iron sulfate, manganese sulfate, and calcium carbonate.

In addition, the medium may contain amino acids, vitamins, and/or appropriate precursors. Specifically, L-amino acids, etc. may be added to the culture medium of the strain. Specifically, glycine, glutamate, and/or cysteine may be added, and if necessary, L-amino acids such as lysine may be added, but are not necessarily limited thereto. No.

The medium or precursor may be added to the culture in a batch or continuous manner, but is not limited thereto.

In the present invention, the pH of the culture can be adjusted by adding compounds such as ammonium hydroxide, potassium hydroxide, ammonia, phosphoric acid, sulfuric acid, etc. to the culture in an appropriate manner during cultivation of the strain. Additionally, during culturing, foam generation can be suppressed by using an antifoaming agent such as fatty acid polyglycol ester. Additionally, to maintain the aerobic state of the culture, oxygen or oxygen-containing gas can be injected into the culture, or to maintain the anaerobic and microaerobic state, nitrogen, hydrogen, or carbon dioxide gas can be injected without the injection of gas.

Another aspect of the present invention provides a food composition containing the above strain.

The “strain” is as described above.

The term "food" in the present invention refers to meat, sausages, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, and alcoholic beverages. , vitamin complexes, health functional foods, and health foods, etc., and include all foods in the conventional sense.

Since the food composition of the present invention can be consumed on a daily basis, it can be expected to have a high immune-boosting effect, so it can be very useful for health promotion purposes.

The above-mentioned health functional food (functional food) is the same term as food for special health use (FoSHU), and is a medicine processed to efficiently exhibit bioregulatory functions in addition to nutritional supply, with high medical effects. It means food. Here, ‘function’ means controlling nutrients for the structure and function of the human body or obtaining useful effects for health purposes, such as physiological effects. The food of the present invention can be manufactured by methods commonly used in the industry, and can be manufactured by adding raw materials and ingredients commonly added in the industry. Additionally, the food formulation can be manufactured without limitation as long as it is a formulation recognized as a food. The food composition of the present invention can be manufactured in a variety of formulations, and unlike general drugs, it is made from food as a raw material and has the advantage of not having side effects that may occur when taking the drug for a long period of time. It is also highly portable, so it can be used as a raw material. The food of the invention can be consumed as an adjuvant to enhance the immune-boosting effect.

The above-mentioned health food refers to food that has a more active health maintenance or promotion effect compared to general food, and health supplement food refers to food for the purpose of health supplementation. In some cases, the terms health functional food, health food, and health supplement are used interchangeably.

Specifically, the health functional food is a food manufactured by adding the compound of the present invention to food materials such as beverages, teas, spices, gum, and confectionery, or by encapsulating, powdering, or suspending it, and consuming it may cause certain health problems. It means bringing about an effect, but unlike regular drugs, it has the advantage of not having any side effects that may occur when taking the drug for a long time since it is made from food.

The food composition may further include a physiologically acceptable carrier. The type of carrier is not particularly limited, and any carrier commonly used in the art can be used.

Additionally, the food composition may contain additional ingredients that are commonly used in food compositions to improve smell, taste, vision, etc. For example, it may include vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, pantothenic acid, etc. Additionally, minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), and chromium (Cr); and amino acids such as lysine, tryptophan, cysteine, and valine.

In addition, the food composition contains preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), disinfectants (bleaching powder, high bleaching powder, sodium hypochlorite, etc.), antioxidants (butylhydroxyanisole (BHA), butylhydroxide) roxitoluene (BHT), etc.), colorants (tar color, etc.), coloring agents (sodium nitrite, sodium nitrite, etc.), bleaching agents (sodium sulfite), seasonings (MSG monosodium glutamate, etc.), sweeteners (dulcine, cyclemate, saccharin) , sodium, etc.), flavorings (vanillin, lactones, etc.), leavening agents (alum, D-potassium hydrogen tartrate, etc.), strengtheners, emulsifiers, thickeners (grease), coating agents, gum base agents, anti-foam agents, solvents, improvers, etc. May contain food additives. The additives can be selected depending on the type of food and used in an appropriate amount.

As an example of the food composition of the present invention, it can be used as a health drink composition, and in this case, it can contain various flavoring agents or natural carbohydrates as additional ingredients, like ordinary drinks. The above-mentioned natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; polysaccharides such as dextrins and cyclodextrins; It may be a sugar alcohol such as xylitol, sorbitol, or erythritol. Sweeteners include natural sweeteners such as thaumatin and stevia extract; Synthetic sweeteners such as saccharin and aspartame can be used. The ratio of the natural carbohydrate may be generally about 0.01 to 0.04 g, specifically about 0.02 to 0.03 g, per 100 mL of the health drink composition of the present invention.

In addition to the above, the health drink composition includes various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, It may contain alcohol or carbonating agent. Additionally, it may contain pulp for the production of natural fruit juice, fruit juice beverage, or vegetable beverage. These ingredients can be used independently or in combination. The ratio of these additives is not very important, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the health drink composition of the present invention.

Another aspect of the present invention provides a pharmaceutical composition containing the above strain.

The “strain” is as described above.

The “pharmaceutical composition” of the present invention may further include pharmaceutically acceptable carriers, excipients, or diluents commonly used in the preparation of pharmaceutical compositions. These pharmaceutically acceptable carriers, excipients, or diluents may be non-naturally occurring. Specifically, the composition is formulated in the form of oral dosage forms such as powders, granules, tablets, solutions, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injection solutions according to conventional methods. can be used In the present invention, carriers, excipients, and diluents that may be included in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, and calcium phosphate. , calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include the extract and its fractions with at least one excipient such as starch, calcium carbonate, It is prepared by mixing sucrose, lactose, and gelatin. In addition to simple excipients, lubricants such as magnesium styrate and talc are also used. Liquid preparations for oral use include suspensions, oral solutions, emulsions, and syrups, and may contain various excipients such as wetting agents, sweeteners, fragrances, and preservatives in addition to the commonly used simple diluents such as water and liquid paraffin. there is. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao, laurin, glycerogenatin, etc. can be used.

The content of the dead cells contained in the pharmaceutical composition of the present invention is not limited as long as the pharmaceutical composition has an immune enhancing effect, but is 0.0001 to 99.9% by weight, more specifically 0.01 to 80% by weight, based on the total weight of the final composition. It may be included in the content.

The pharmaceutical composition of the present invention can be administered in a pharmaceutically effective amount. As used herein, the term “pharmaceutically effective amount” refers to an amount sufficient to treat or prevent a disease with a reasonable benefit/risk ratio applicable to medical treatment or prevention. This refers to the amount, and the effective dose level is the severity of the disease, the activity of the drug, the patient's age, weight, health, gender, the patient's sensitivity to the drug, the administration time, route of administration, and excretion rate of the composition of the present invention used. Treatment period , may be determined according to factors including the composition of the present invention and the drugs used in combination or simultaneous use, and other factors well known in the medical field. The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. And it can be administered single or multiple times. It is important to consider all of the above factors and administer the amount that will achieve the maximum effect with the minimum amount without side effects.

The dosage of the pharmaceutical composition of the present invention is, for example, 0.1 to 500 mg/kg of body weight per day to animals, including humans, but is not limited thereto. The frequency of administration of the composition of the present invention is not particularly limited, but may be administered once a day or divided into multiple doses. The above dosage does not limit the scope of the present invention in any way.

Another aspect of the present invention provides a method for enhancing immunity comprising administering the pharmaceutical composition to an entity other than a human.

The term "individual" in the present invention refers to all animals such as rats, mice, and livestock, including humans, that require immunity enhancement. As a specific example, it may be a mammal, including humans.

As used herein, the term “administration” refers to the act of introducing the pharmaceutical composition into an individual by an appropriate method.

Another aspect of the present invention provides a feed composition containing the above strain.

The “strain” is as described above.

In the present invention, the term "feed" means any natural or artificial diet, meal, etc., or a component of the meal, for or suitable for eating, ingestion, and digestion by animals.

The type of feed is not particularly limited, and feed commonly used in the art can be used. Non-limiting examples of the feed include plant feed such as grains, roots and fruits, food processing by-products, algae, fiber, pharmaceutical by-products, oils and fats, starches, gourds or grain by-products; Examples include animal feeds such as proteins, inorganic substances, fats and oils, minerals, fats and oils, single-cell proteins, zooplanktons or food. These may be used alone or in combination of two or more types.

Another aspect of the present invention includes the step of heat-treating the Bacillus velezensis GV1 strain deposited under deposit number KCTC 15222BP at 100°C to 150°C for 5 to 30 minutes to kill the strain. A method for producing dead cells is provided.

The “Bacillus” and “strain” are as described above.

In the present invention, the dead cells are Bacillus velegensis GV1 strain deposited under the deposit number KCTC 15222BP or a culture of the strain at 100 ℃ to 150 ℃, specifically 105 ℃ to 140 ℃, more specifically 110 ℃ to 130 ℃ for 5 days. It may be manufactured through heat treatment for minutes to 30 minutes, specifically 7 minutes to 25 minutes, and more specifically 10 minutes to 20 minutes.

The Bacillus velezensis GV1 strain deposited under the deposit number KCTC 15222BP of the present invention can exhibit acid resistance, bile resistance, and gluten-decomposing activity, and can be widely used in food, pharmaceutical, and feed compositions.

Figure 1 is a graph showing the growth rate according to the acid resistance and pH of the strain.
Figure 2 is a graph showing the resistance of the strain to 0.5%-Bile salt.
Figure 3 shows the results showing the morphological characteristics and gluten decomposition ability of the strain.

Hereinafter, the present invention will be described in more detail through examples and experimental examples. However, these examples and experimental examples are for illustrative purposes only and the scope of the present invention is not limited to these examples and experimental examples.

Experimental Example 1. Isolation and identification of Bacillus velezensis GV1

It was isolated and identified from ginseng vinegar.

Bacillus velezensis GV1 strain, a lactic acid bacteria derived from ginseng vinegar, was obtained.

The Bacillus velezensis GV1 strain was deposited on December 5, 2022 at the Korea Research Institute of Bioscience and Biotechnology Biological Resources Center, a depository under the Budapest Treaty, and was given the accession number KCTC 15222BP. The strain was confirmed to have the rDNA base sequence of SEQ ID NO: 1.

Experimental Example 2. Preparation of dead cells

Bacillus velezensis GV1 was cultured in MRS (Man Rogosa Sharpe) medium for 24 hours using a shaking incubator (RPM180, 37°C), then centrifuged (8000 x g, at 4°C). After centrifugation for 10 minutes, the supernatant was removed, and the remaining residue was washed twice using PBS. After setting the OD value to 1 at 600 nm, heat treatment was performed at 121°C for 15 minutes, and the freeze-dried sample was suspended at 10 mg/mL and subjected to sonication for 30 minutes to obtain dead cells.

As a comparison group, the Bacillus belegensis kh2-2 strain was used, and kh2-2 was isolated and identified from salted squid, and specifically, dead cells were prepared using the method described in the paper (Food Research International Volume 152).

Example 1. Confirmation of acid resistance activity of Bacillus velegensis GV1 strain

The growth of Bacillus velgensis GV1 and the comparative strain Bacillus velgensis kh2-2 was monitored by time in a culture medium adjusted to pH 1 to pH 12 after 0 and 12 hours, and then the growth rate was measured at OD ₆₀₀ ( Figure 1).

As a result, it was confirmed that the growth ability of Bacillus velegensis GV1 was excellent in pH 2 medium, the number of bacteria increased significantly over time, and it was confirmed that growth was not inhibited under strong acid conditions. On the other hand, it was confirmed that Bacillus belegensis kh2-2 did not proliferate. Through this, it was found that Bacillus velegensis GV1 has acid resistance.

Example 2. Confirmation of bile-resistant activity of Bacillus velegensis GV1 strain

Bile salt of Bacillus velegensis GV1 and the comparative strain Bacillus velegensis kh2-2 were raised from 0.5% to 2% and the growth of the strains was monitored by time after 0 and 50 hours, and then the growth rate was measured at OD ₆₀₀ ( Figure 2).

As a result, the strain showed excellent growth ability in medium supplemented with 0.3% or 0.5% bile. Through this, it was found that Bacillus velegensis GV1 has bile resistance.

Example 3. Confirmation of gluten decomposition activity of Bacillus velegensis GV1 strain (Gluten Hydrolysis Tests)

5 ul of the bacterial culture (10 ⁸ CFU/ml) was plated on a solid medium containing 1% gluten and cultured at 37 degrees for 48 hours. The plate was then stained with Coomassie Blue Brilliant R-250 dye (Sigma-Aldrich) for 1 minute and then removed. It was stored (TR) over night with washing solution (250 mL/L ethanol, 50 mL/L acetic acid). Decomposition of gluten was confirmed by observing a clear zone generated as gluten was decomposed around the colony. Specifically, gliadin was comparatively measured using RIDASCREEN Gliadin competitive (R7021) and RIDASCREEN Gliadin (R7001), analysis kits for measuring the amount of gluten (FIG. 3).

As a result, it was confirmed that the unstained area was the place where the gluten was decomposed, and that a large unstained ring was generated around the Bacillus velegensis GV1 colony. Through this, it was confirmed that the Bacillus velegensis GV1 of the present invention has excellent gluten decomposition ability. Could know.

From the above description, those skilled in the art to which the present invention pertains will understand that the present invention can be implemented in other specific forms without changing its technical idea or essential features. In this regard, the embodiments described above should be understood in all respects as illustrative and not restrictive. The scope of the present invention should be construed as including the meaning and scope of the patent claims described below rather than the detailed description above, and all changes or modified forms derived from the equivalent concept thereof are included in the scope of the present invention.

Korea Institute of Bioscience and Biotechnology Biological Resources Center KCTC15222BP 20221205

Claims (8)

Bacillus velezensis GV1 strain deposited with deposit number KCTC 15222BP.
The method of claim 1, wherein the strain is
i) acid resistance; ii) endobiliary; and iii) a strain characterized by having at least one activity from the group consisting of gluten-decomposing activity.
The strain of claim 1 is a dead cell.
The method of claim 3, wherein the dead cells are prepared by heat-treating the Bacillus velegensis GV1 strain deposited under the accession number KCTC 15222BP or a culture of the strain at 100°C to 150°C for 5 to 30 minutes, strain.
A food composition comprising the strain of claim 1.
A pharmaceutical composition comprising the strain of claim 1.
A feed composition comprising the strain of claim 1.
A method for producing dead cells, comprising the step of killing the Bacillus velezensis GV1 strain deposited under the accession number KCTC 15222BP by heat treating the strain at 100°C to 150°C for 5 to 30 minutes.