KR20190142263A - Lactobacillus plantarum MB-0601 strain having gluten degradation activity from salted seafood and uses thereof - Google Patents

Abstract

The present invention relates to a Lactobacillus plantarum MB-0601 strain having gluten degradation activity isolated from salted seafood, and uses thereof. The Lactobacillus plantarum MB-0601 strain according to the present invention can be used to prepare popular gluten-free processed foods such as noodles, udon, pizza and bread, and thus can be very useful to people who have not consumed grain flour processed foods due to celiac disease or gluten allergy caused by gluten intolerance.

Description

Lactobacillus plantarum MB-0601 strain having gluten degradation activity from salted seafood and uses according to Lactobacillus plantarum MB-0601 strain having gluten resolution isolated from salted fish

The present invention relates to a Lactobacillus plantarum MB-0601 strain having gluten resolution isolated from salted fish and its use.

Flour is one of the most important raw materials in the Korean food industry. The main use is noodle, confectionery, and bakery. In 2001, the flour production amounted to 17.77 million tons, of which 37.7% was used for confectionery and 21.9% for confectionery and bakery. The flour produced in Korea is strong flour, gravity flour, and flour, and there are mixed flour and wholemeal flour produced since 1986. The mixed flour belongs to gravity flour for noodles. The whole wheat flour is a crushed whole wheat and is used for special baking and confectionery. The strong wheat flour is mainly made of Dark Northern Spring (DNS) in the United States, and Canada's CWRS (Canada Western Red Spring) since 1991 It began to be used for baking.

Gravity flour is produced by milling a mixture of white wheat and hard red winter in the same proportions, and is mainly used for noodle and home multipurpose flour. Recently, Australian wheat is used as a noodle flour, and Australian wheat is mainly composed of Australian Standard White, Australian White and Australian Hard. As the preference for ASW, a major Australian wheat species, has increased significantly with the diversification of quality and the quality of quality, it is increasing year by year. In the case of flour, there have been hundreds of years of systematic research and industrialization, producing hundreds of products optimized for each food. In Western society, where wheat is a staple, most wheat products are bread, cookies, noodles, fries and pizza. It has been developed in the back, and most of the processed wheat products in the processing mainly carbohydrates. In recent years, as the food culture has gradually become westernized, rice is a staple food, but the annual consumption of rice per capita has declined by 23% from 992 kg in 1998 to 774 kg in 2009. The trend is increasing. However, grains such as barley and wheat contain gluten, an insoluble protein, which makes it difficult to digest itself and may cause gluten intolerance.

Gluten is a by-product of wheat starch production and is an insoluble protein in water. It is composed mainly of gliadin and glutenin. Gliadin has a molecular weight of 30,000 to 80,000 Da, and glutenin is millions of Da. It has a large molecular weight. In particular, glutamic gliadin is soluble in alcohol, but glutenin is insoluble, but glutenin plays an important role in baking. In other words, when wheat flour is mixed with water, gliadin and glutenine combine to form gluten. Among the gluten proteins, gliadin is viscous and extensible, and glutenin has elasticity. These two proteins occupy different proportions depending on the type of flour, and this is a unique property that only flour has. For example, in the wheat flour used to make bread, 65% of gliadin and 35 of glutenine are used. And wheat flour, which is used to make cakes, contains 75% of gliadin and 25% of glutenine. Its role is related to the volume of the bread and is responsible for the skeletal formation of dough and the gases produced during fermentation. It has the function to hold. Gluten is indigestible on its own, and gluten intolerance can adversely affect your skin, nervous system, immune system, stamina, joints, teeth, behavior and mood, often known as gluten allergy, but the real name is gluten-sensitive bowel disease. 'to be.

Intolerance is a reaction that our body fails to absorb nutrients when it enters the body. A typical example is gluten intolerance symptoms, which may be accompanied by digestive problems such as diarrhea, abdominal pain, constipation, and bloating. Gluten-sensitive bowel disease is caused by the absence or lack of an enzyme that digests gluten.Ingestion of gluten can irritate the intestines and weaken the process of the intestinal mucosa, making it impossible to absorb nutrients. It is a hereditary disease that occurs only in people who do not have gluten digestive enzymes.

As is commonly known, Celiac disease, a gluten-resistant disease, is an allergic disease that occurs in the small intestine and is caused by gluten susceptibility to intestinal nutrient absorption, which usually begins in infancy but rarely appears as an adult. In some cases. At least one out of every 133 Americans have celiac disease, but more than three times more people are suspected of having gluten sensitivity, and have recently developed rapidly among Europeans (especially Northern Europe) and Latin Americans. Blacks and Asians are also on the rise.

Meanwhile, Korean Patent Application Publication No. 2017-0068894 discloses' Bisella Confusa CJE-0201 strain having its gluten-degrading ability isolated from salted fish and its use ', and Korean Patent Application Publication No. 2015-0083899 discloses' within sourdough. Microencapsulated bacterial groups and glutamate sourdough for gluten breakdown of the present invention are disclosed, but as in the present invention, 'Lactobacillus plantarum MB-0601 strain having gluten degradability isolated from salted fish and its use 'Is not known at all.

The present invention is derived from the above requirements, the present inventors search for lactic acid bacteria that can decompose gluten causing celiac disease from domestic fermented foods, and among them, the lactobacillus having a gluten degrading ability to analyze the gluten resolution It was. As a result, the fermentation of gluten-containing grains (wheat, rye, barley, oats, etc.) using the Lactobacillus plantarum MB-0601 strain selected in the present invention confirms that the gluten present in the grain flour can be decomposed. It was confirmed that the Lactobacillus plantarum MB-0601 strain produced a gluten degrading enzyme having a molecular weight of about 67,000 Da. Therefore, the gluten-free processed foods such as noodles, udon, pizza, and bread may be prepared using the gluten-degraded grain powder prepared by fermentation as described above, and thus, the present invention is expected to be very useful in the related food industry. Was completed.

In order to achieve the object of the present invention, the present invention provides a Lactobacillus plantarum MB-0601 strain which is isolated from salted fish and has a gluten resolution of accession number KCTC 13532BP.

The present invention also provides a microbial preparation for gluten decomposition comprising the strain or its culture as an active ingredient.

In another aspect, the present invention provides a method for producing a gluten-free cereal flour fermentation product comprising the step of treating the strain or its culture solution with cereal flour and then fermenting it to break down the gluten present in the cereal flour. .

The present invention also provides a gluten-free grain flour fermented product prepared by the above method.

The present invention also provides a gluten-free processed food prepared from the gluten-free cereal flour fermented product.

Gluten-reduced cotton products fermented with Lactobacillus plantarum MB-0601 strain (KCTC 13532BP) according to the present invention improve gastrointestinal digestion by promoting digestive enzyme-related gastrin secretion through animal experiments. Efficacy was confirmed. Therefore, the Lactobacillus plantarum MB-0601 strain isolated in the present invention has excellent gluten-degrading ability, so that gluten-containing grains are ingested by decomposing gluten present in wheat starch or cereals through grain fermentation using the strain. It can improve gluten-sensitive bowel disease known as allergies.

Therefore, using the Lactobacillus plantarum MB-0601 strain according to the present invention can produce a popular gluten-free processed foods such as noodles, udon, pizza and bread, celiac disease caused by gluten intolerance (Celiac disease) Or it may be very useful to those who could not eat the grain flour processed food due to gluten allergy.

Figure 1 shows a culture photograph of the Lactobacillus plantarum MB-0601 strain isolated in the present invention.
Figure 2 shows the 16s rDNA sequence (SEQ ID NO: 1) for the genetic identification of the Lactobacillus plantarum MB-0601 strain isolated in the present invention.
Figure 3 shows the results of screening proteolytic enzyme activity of the dialysate obtained through the ammonium sulfate precipitation from the culture supernatant of the Lactobacillus plantarum MB-0601 strain isolated in the present invention.
Figure 4 shows the result of purifying the dialysate having protease activity selected by the ammonium sulfate precipitation from the culture supernatant of the Lactobacillus plantarum MB-0601 strain isolated in the present invention using the DEAE column chromatography method.
5 is a result of confirming the gluten degrading activity peak through the first (A) and the second (B) purification by performing affinity column chromatography method on the active fraction obtained by using the DEAE column chromatography method.
Figure 6 is a result of measuring the molecular weight of the gluten degrading enzyme produced by the Lactobacillus plantarum MB-0601 strain isolated in the present invention.
Figure 7 shows the feces of digestive disorder-induced mice by dextran sulfate sodium.
Figure 8 confirms the results on the enhancement of amylase secretion digestion enzyme when ingesting fermented noodles using the Lactobacillus plantarum MB-0601 strain isolated in the present invention.
Figure 9 confirms the results of the gastrin secretion of digestive hormones when the fermented cotton intake using the Lactobacillus plantarum MB-0601 strain isolated in the present invention.

In order to achieve the object of the present invention, the present invention provides a Lactobacillus plantarum MB-0601 strain which is isolated from salted fish and has a gluten resolution of accession number KCTC 13532BP.

Therefore, the present inventors searched for lactic acid bacteria that can decompose gluten from fermented foods of domestic fermented food, and analyzed the gluten decomposition ability by selecting the lactic acid bacteria having excellent gluten resolution among them. The final secured lactic acid bacteria was deposited on May 28, 2018 by the Korea Biotechnology Research Institute Gene Bank (KCTC, Korean Collection for Type Cultures) to complete the present invention by receiving the accession number KCTC 13532BP.

In addition, the Lactobacillus planta MB-0601 strain of the present invention can produce a gluten degrading enzyme having a molecular weight of about 67,000 Da, but is not limited thereto.

In the present invention, celiac disease caused by gluten intolerance is an enterophatic, chronic, immune-mediated type caused by the ingestion of gluten, a "storage" protein naturally contained in some grains. It is an inflammatory disease. These food intolerances include the consumption of wheat gluten, spells (a type of flour), kamut and spellas (a type of flour), barley, rye, rye (a hybrid of wheat and rye) and their derivatives of typical protein fractions. It affects genetically predisposed people with an immune system that responds in an abnormal way. Some individuals are intolerant of oat proteins.

In the present invention, the term "gluten" technically refers to a simple prolaminic (proline-rich) protein called "gliadin" and a glutenic called "glutenin", which is a protein of the aforementioned grains. A combination of (glutelinic) (glutamine-rich) proteins. With respect to celiac disease, "gluten" is used to collectively refer to the proteins included in the grain, including the various protein fractions that form gluten, of which gliadin is considered the most harmful. The possibility of preventing or treating celiac disease has not existed until now, so strictly eating gluten-free throughout life is the only remedy to ensure perfect health for people with celiac disease.

The present invention also provides a microbial agent for gluten decomposition comprising the strain of the present invention or a culture thereof as an active ingredient. The strain is preferably Lactobacillus plantarum , more preferably Lactobacillus plantarum MB-0601 strain (KCTC 13532BP).

In addition, the present invention provides a method for producing a microbial agent for gluten decomposition comprising culturing the strain.

The strain is preferably Lactobacillus plantarum MB-0601 strain (KCTC 13532BP). The method of culturing the strain of the present invention may use a method commonly used in the art.

In addition, the lactic acid bacteria strain and lactic acid bacteria culture solution provided in the present invention, granules, solids and the like can be formulated and supplied according to the type and purpose of use.

In another aspect, the present invention provides a method for producing a gluten-free cereal flour fermentation product comprising the step of treating the strain or its culture solution with cereal flour and then fermenting it to break down the gluten present in the cereal flour. .

In the method according to an embodiment of the present invention, the grains may be one or more selected from the group consisting of wheat, rye, barley, oats, spells and kamut, but is not limited thereto.

The present invention also provides a gluten-free grain flour fermented product prepared by the above method. The "gluten free grain flour fermented product" is interpreted to include grain flour fermented with gluten removed mostly in addition to grain flour fermented with gluten completely removed.

The present invention also provides a gluten-free processed food prepared from the gluten-free cereal flour fermented product. The gluten free processed foods may be, but are not limited to, noodles, udon, confectionery, ramen, pizza, or bread made from gluten free grain flour fermented products.

Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited to the following examples.

Example  1. Securing gluten lactic acid bacteria

In order to separate lactobacillus with gluten degrading ability from various salted fish, various salted fish salts are chopped aseptically, 1g of which is suspended in 9 ml of sterile saline solution (0.85% NaCl) and diluted stepwise to contain 1% of gluten. Each was plated on MRS solid medium. After incubation at 30 ° C. for 48 hours, only microorganisms forming a clear zone generated by decomposing gluten around colonies appeared were isolated. Subsequently, the microorganisms purely separated from the salted fish were plated in an MRS solid medium and incubated at 30 ° C., and the colonies formed were suspended in 20% glycerol solution and then stored in a -80 ° C. freezer, and used to search for gluten degrading lactic acid bacteria.

Example  2. Genetic Identification of Lactic Acid Bacteria Isolated from Salted Fish

For genetic identification of the isolated strain, 2 ml of the culture was centrifuged at 4,000 rpm for 10 minutes by inoculating in MRS liquid medium and then subcultured at 37 ° C. for 18 hours. The cell precipitate obtained by centrifugation was washed three times with 0.85% NaCl. After washing, 0.5 ml of lysozyme (10 mg / ml) was added and treated at 37 ° C. for 1 hour. After adding 20 μl of protease K, 25 μl of protease K and 25 μl of 10% sodium dodecyl sulfate (SDS), the result was treated for 30 minutes in a 60 ° C. water bath, followed by RNA degradation enzyme. 1 μl of phosphorus Rn was added and treated at 37 ° C. for 1 hour. Suspension was added with the same amount of Phenol-Chloroform-Isoamyl alcohol in a ratio of 25: 24: 1, followed by centrifugation at 4 DEG C for 5 minutes at 14,000 rpm, and only the supernatant was taken. Then, this process was repeated twice. One half of the amount of supernatant was added 3M ammonium acetate (pH 4.8) and twice the amount of 100% alcohol (alcohol) and left at -20 ° C for 1 hour. After confirming the cell precipitate by centrifugation at 4 ℃ for 5 minutes at 3,000 rpm, after removing the supernatant completely 1ml of 70% ethanol (ethanol) was added again centrifuged at 4 ℃ for 5 minutes at 3,000 rpm. After removing the supernatant, dried and suspended in TE buffer or distilled water to separate the DNA. Forward primer (27f, 5'-AGA GTT TGA TCM TGG CTC AG-3 ': SEQ ID NO: 2) and reverse primer (1492r, 5'-GGT TAC CTT TGT TAC GAC TT -3' to amplify 16s rRNA of microorganism : SEQ ID NO: 3) was used. PCR was performed after mixing 17 μl of distilled water, 1 μl forward primer, 1 μl reverse primer, and 1 μl DNA to the PCR premix (Bioneer, Cat No. K-2012). PCR conditions were repeated 5 times at 94 ℃ for 5 minutes, 1 minute at 94 ℃, 40 seconds at 62 ℃, 40 times at 72 ℃ 40 times, the reaction was terminated in 7 minutes at 72 ℃. PCR reaction products were confirmed by electrophoresis on a 0.8% agarose gel (agarose gel). The final nucleotide sequence (SEQ ID NO: 1, Figure 2) was analyzed and compared with the database to identify the lactic acid bacteria, which is shown in Figure 2.

As a result, as shown in Figures 1 and 2, the strain isolated by the present invention was identified as Lactobacillus plantarum , Lactobacillus plantarum ( Lactobacillus plantarum ) MB-0601 by the Korea Biotechnology Research Institute It was deposited on 28 May 2018 at BRC (Accession No. KCTC 13532BP).

Example  3. Lactobacillus Planta Room  Gluten Resolution of MB-0601 Strain

Lactobacillus plantarum MB-0601 strain, a gluten fermented lactic acid bacterium, was applied to a process for producing cotton to evaluate gluten degradability in wheat flour. Experimental method was kneaded by adding 500 g of water to 1 kg of gravity flour and adding gluten fermented lactic acid bacteria (prototype). At this time, the experimental group 1, 0.6 g (0.06%) was added to the group added 1.5 g (0.15%) of gluten fermented lactobacillus to the dough and the experimental group 2, 0.3 g (0.03%) was added to the experimental group 3 Recorded as. After kneading the flour, the gluten decomposition ability was evaluated through the aging process at 30 ℃. Gluten content evaluation method was analyzed using the separation of food proteins (separation of gluten from flour) of the National Food Science Institute of Rural Development Administration. The gluten resolution according to the addition of lactic acid bacteria was shown in Table 1 below.

Evaluation of Gluten Degradation Rate of Udon Noodles with Gluten Fermented Lactic Acid Bacteria Lactobacillus added amount Dry Gluten (g) % Decomposition Average(%) Control 1.85 0.00 0 Experiment group 1 0.49 73.51 67.027 0.73 60.54 Experiment group 2 0.69 62.70 69.730 0.43 76.76 Experiment group 3 0.75 59.46 58.108 0.8 56.76

Example 4 Securing Glutenase Enzyme Secreted by Lactobacillus Planta Room MB-0601 Strain

1) Ammonium Sulphate Precipitation

Lactobacillus plantarum MB-0601 strains were inoculated 1% in 1 L of MRS medium adjusted to pH 7.0 and incubated for 24 hours. The supernatant obtained by removing the cells through centrifugation for 20 minutes at 6,000 rpm was added to the culture medium in a 400mL centrifuge tube was used for the experiment. For the isolation and purification of gluten degrading enzymes, ammonium sulfate was first added at saturated concentrations of 0-20%, 20-40%, 40-60%, 60-80% and 80-100% The precipitated protein was dissolved in 20 mM Tris-HCl, pH 7.0 buffer containing 10 mM CaCl ₂ and dialyzed at 4 ° C. for 12 hours with the same buffer. 100 μl of each dialysate was used as a coenzyme, and the activity was measured by agar diffusion method on MRS solid medium containing 1.5% skim milk. As a result, as shown in FIG. 3, the protease activity was measured only in the 40-60% saturation section of ammonium sulfate, and thus the above section was selected for proteolytic enzyme purification. Proteins recovered from ammonium sulfate (40-60%) were dialyzed with 20 mM Tris-HCl (pH 7.0) buffer containing 20 mM CaCl ₂ as an initial buffer of ion exchange chromatography.

2) DEAE column chromatography

Purification experiments were carried out on the coenzyme obtained by the ammonium sulfate precipitation using DEAE-sepharose column chromatography. At this time, NaCl gave a concentration gradient of 0 ~ 1M. 100 μl of each fraction was aliquoted, and the activity of the MRS solid medium containing 1.5% skim milk was measured by agar diffusion method. Also, the enzyme activity and protein content containing 1% casein were analyzed. As shown in FIG. 4, as the NaCl content in the column increases, the amount of the eluted enzyme protein gradually increases. The enzyme activity fraction was eluted in 49-55 fractions and the enzyme activity was maximal in 52 fractions with the highest protein content. Only 49-55 fractions were collected, and the enzyme solution was dialyzed three times at 2 hour intervals in distilled water, and then concentrated in an Amicon concentrator (Amicon) using an Amicon membrane (M.W.1,000). The concentrated sample was dissolved in 20mM Tri-HCl buffer and used as a sample for the next experiment.

3) Affinity column chromatography

The active fractions obtained from the DEAE-sepharose column chromatography experiments were collected to perform affinity column chromatography experiments. 100 μl of each fraction was aliquoted, and the activity of the MRS solid medium containing 1.5% skim milk was measured by agar diffusion method. Also, the enzyme activity and protein content of 1% casein were analyzed. As a result, as shown in FIG. 5A, a peak having gluten degradation activity in the 9-16 fraction was confirmed. 9-16 fractions obtained from different types of protein peaks were collected and subjected to a second affinity column chromatography experiment to collect an active fraction of a single peak (FIG. 5B). The specific activity of the finally purified enzyme was 49.4 unit / mg, yield was 4.9%, and purity was 17.4 times.

4) Determination of activity of gluten degrading enzyme

Enzyme activity was measured using a tyrosine assay (Folin, 1927, J. Biol. Chem., 73, 627-650). 100 μl of sample solution was pre-incubated for 5 minutes in a shaker at 40 ° C., followed by 900 μl of 1% substrate solution for 60 minutes at the same temperature, and 1,000 μl of 10% TCA solution was added to stop the enzyme reaction. . The non-hydrolyzed proteins were left at room temperature for 10 minutes and then removed by centrifugation at 10,000 rpm for 10 minutes. 200 μl of the supernatant and 800 μl of 0.55 M Na ₂ CO ₃ were added, mixed, and left to stand for 15 minutes. 200 μl of 1.0N Folin Ciocalteu's phenol solution was added thereto, mixed, left for 30 minutes, and absorbance was measured at 660 nm. The enzyme activity unit was defined as 1 unit of 1 μM of tyrosine produced by 1.0 mL of sample solution, and the units were calculated from a standard curve prepared using tyrosine.

Enzymatic Activity of Purifying Glutenase Purification steps Unit Total protein (mg) Specific activity ⁽¹⁾
(U / mg) Yield ⁽²⁾
(%) Purification fold Supernatant 38,456 13,564 2.9 100 One Ammonium sulfate 16,877 1,856 9.1 43.9 3.2 DEAE-sepharose 6,587 448 14.7 17.1 5.2 1st Affinity
chromatography 2,566 66 38.8 6.7 13.7 2nd Affinity
chromatography 1,878 38 49.4 4.9 17.4

1) Specific activity = total activity / total protein.

2) Yield = (total activity) / (total activity of supernatant) × 100.

5) Determination of Molecular Weight of Glutenase

The molecular weight of the final purified gluten degrading enzyme was measured according to Laemmli's method (1970, Nature, 227, 680-685) and the molecular weight calculation was determined as the ratio of the elution volume of protease to void volume (Ve / Vo). Standard curves for molecular weight measurements were prepared with standard proteins cytochrome C (MW 12,400 Da), carbonic anhydrase (MW 29,000 Da), albumin (MW 66,000 Da), alcohol dehydrogenase (MW 150,000 Da), and apoferritin (MW 443,000 Da). . The standard curve was created from the ratio of the elution volumes of each standard protein. At this time, the standard curve was prepared by exponential equation, and the standard curve was y = 1E + 07e ^-2.932x , and the reliability was R² = 0.9968. The molecular weight was calculated by introducing the elution volume of the purified gluten degrading enzyme into the exponential formula of the standard curve thus prepared, and the formula for measuring the molecular weight of the gluten degrading enzyme produced by the Lactobacillus plantarum MB-0601 strain is as follows.

One.

2.

3.

Vt = total bed volume, total volume in column

Vo = void volume, spatial volume between gel particles in DEAE column chromatography

Vi = inner volume, volume of gel particles containing water

Ve = elution volume, volume until gluten degrading enzyme is eluted

Vt = Vo + Vi

K = distribution coefficient, volumetric volume / static volume of protein transported inside the gel particle

Kav = mean distribution coefficient

Vm = volume of mobile phase = Vo

Vs = volume of stationary phase = Vi

Vr = volume with gluten degrading enzyme = Ve

Therefore, as a result of measuring the molecular weight of the gluten degrading enzyme produced by the Lactobacillus plantarum MB-0601 strain through the above formula, the elution volume ratio (Ve / Vo) of the gluten degrading enzyme was 1.8 and the molecular weight was about 67,000 Da. (FIG. 6).

Example  5. Analysis of Digestive Effect of Gluten Fermented Noodle Product Using Animal Model

Digestive efficacy in animals was confirmed for cotton products fermented with Lactobacillus plantarum MB-0601 strain. After randomly grouping a total of 60 four-week-old C57BL / 6 male mice, each test group was divided in the manner as shown in Table 3 below to perform the efficacy test.

Design of Animal Experiment group UT Control Gluten-free Low concentration Medium concentration High concentration % kcal from fat 18 18 18 18 18 % kcal from wheat - 60 60 60 60 Gluten Cotton (%, w / w) - 30 27 18 0 Gluten Free Cotton
(% .w / w) - 0 3 12 30 Number of animals (mouse) 12 12 12 12 12 gender cock cock cock cock cock Feeding period (week) 8 8 8 8 8

1) UT (untreated)

2) normal feed + normal wheat flour 30% (w / w) (CTL; control, negative control)

3) General Feed + Normal Flour 27% + Gluten-Free Flour 3% (low concentration)

4) General Feed + Wheat Flour 18% + Gluten-Free Flour 12% (Medium)

5) General Feed + Gluten-Free Flour 30% (High Concentration)

After the end of the period of purification for one week, all groups except the UT group received negative 5% dextran sulfate sodium (DSS) in sterile distilled water for one week. DSS is 2-5% of the negative induction of digestive disorders, which can be seen through weight and bowel movement (Fig. 7).

Amylase, a gluten-related digestive enzyme in vivo, is an enzyme that hydrolyzes glycoside bonds of polysaccharides, and may increase digestion. When pancreatic amylase was measured by decomposing pancreatic tissue, the negative control group showed 0 ± 0.0005 mU / ml. Gluten-free flour low-dose group was 0.004 ± 0.001 mU / ml, medium-dose group was 0.005 ± 0.002 mU / ml and high-dose group was 0.008 ± 0.002 mU / ml. . This increased pancreatic glycolysis activity by gluten-free flour (both p <0.001) (FIG. 8).

In addition, gastrin among the digestive enzymes and related hormones in the blood is a hormone secreted by gastric anterior G cells, which promotes secretion of gastric juice and helps gastrointestinal movement. Blood gastrins were measured using a commercial diagnostic kit. As a result, the negative control group showed 37.33 ± 9.06 pg / ml and the gluten-free flour high concentration was 59.64 ± 18.48 pg / ml, which was 60% higher in the gluten-free flour concentration than the negative control ( p <0.01) . It was confirmed that gluten-free flour supplementation may improve gastrointestinal digestion by promoting gastrin secretion (FIG. 9).

Korea Research Institute of Bioscience and Biotechnology KCTC13532BP 20180528

<110> Korea Research Institute of Bioscience and Biotechnology          Microbiome Co., Ltd. <120> Lactobacillus plantarum MB-0601 strain having gluten degradation          activity from salted seafood and uses <130> PN19215 <160> 3 <170> KopatentIn 2.0 <210> 1 <211> 1017 <212> DNA <213> Lactobacillus plantarum MB-0601 <400> 1 tgcagtcgaa cgaactctgg tattgattgg tgcttgcatc atgatttaca tttgagtgag 60 tggcgaactg gtgagtaaca cgtgggaaac ctgcccagaa gcgggggata acacctggaa 120 acagatgcta ataccgcata acaacttgga ccgcatggtc cgagcttgaa agatggcttc 180 ggctatcact tttggatggt cccgcggcgt attagctaga tggtggggta acggctcacc 240 atggcaatga tacgtagccg acctgagagg gtaatcggcc acattgggac tgagacacgg 300 cccaaactcc tacgggaggc agcagtaggg aatcttccac aatggacgaa agtctgatgg 360 agcaacgccg cgtgagtgaa gaagggtttc ggctcgtaaa actctgttgt taaagaagaa 420 catatctgag agtaactgtt caggtattga cggtatttaa ccagaaagcc acggctaact 480 acgtgccagc agccgcggta atacgtaggt ggcaagcgtt gtccggattt attgggcgta 540 aagcgagcgc aggcggtttt ttaagtctga tgtgaaagcc ttcggctcaa ccgaagaagt 600 gcatcggaaa ctgggaaact tgagtgcaga agaggacagt ggaactccat gtgtagcggt 660 gaaatgcgta gatatatgga agaacaccag tggcgaaggc ggctgtctgg tctgtaactg 720 acgctgaggc tcgaaagtat gggtagcaaa caggattaga taccctggta gtccataccg 780 taaacgatga atgctaagtg ttggagggtt tccgcccttc agtgctgcag ctaacgcatt 840 aagcattccg cctggggagt acggccgcaa ggctgaaact caaaggaatt gacgggggcc 900 cgcacaagcg gtggagcatg tggtttaatt cgaagctacg cgaagaacct taccaggtct 960 tgacatacta tgcaaatcta agagattaga cgttcccttc ggggacatgg atacagg 1017 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 2 agagtttgat cmtggctcag 20 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 3 ggttaccttt gttacgactt 20

Claims (6)

Lactobacillus plantarum MB-0601 strain isolated from salted fish and having a gluten resolution of accession number KCTC 13532BP. Microbial agent for gluten decomposition comprising the strain of claim 1 or a culture thereof as an active ingredient. A method of producing a gluten-free cereal flour fermentation product comprising the step of treating the strain of claim 1 or its culture solution in cereal flour and then fermenting the gluten present in the cereal flour. 4. The gluten-free cereal flour fermentation according to claim 3, wherein the grains are at least one selected from the group consisting of wheat, rye, barley, oats, spells and kamuts. Method of producing water. Gluten-free cereal flour fermentation prepared by the method of claim 3 or 4. A gluten free processed food prepared from the gluten free grain flour fermentation product of claim 5.

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