KR101977255B1 - Leuconostoc mesenteroides MKSR,a health functional food and food additive for preventing or improving diabetes comprising the same - Google Patents

Abstract

The present invention relates to a Leuconostoc mesenteroides MKSR strain deposited with Accession number KCTC18665P, and functional health food and a food additive for preventing or ameliorating diabetes comprising the same.

Description

(Leuconostoc mesenteroides MKSR) deposited with the deposit number KCTC18665P, a health functional food for preventing or improving diabetes comprising the same, and a food additive (Leuconostoc mesenteroides MKSR, a health functional food and food additive for preventing or improving diabetes comprising the same}

The present invention relates to a strain of Leuconostoc mesenteroides MKSR (MKSR) deposited with deposit number KCTC18665P, a health functional food for preventing or improving diabetes and a food additive comprising the same.

Diabetes is the most common disease of endocrine disorders caused by insulin's failure to function normally. Globally, diabetes is estimated to be 347 million people, or about 5% of the world's population, with about 3.4 million people dying of fasting hyperglycemia. Moreover, diabetes is accompanied by long-term complications such as arteriosclerosis, hypertension, cerebrovascular infarction, diabetic nephropathy, diabetic retinopathy, maculopathy, eczema, gangrene and oral disease. WHO reports that the prevalence of diabetes is steadily increasing, Is expected to be the seventh in mortality. According to the American Diabetes Association, annual healthcare costs for diabetics reach $ 132 billion (2002), and type 2 diabetes, which accounts for 90% of diabetic patients, is growing rapidly due to weight gain and lack of exercise . In Korea, due to socioeconomic development, diabetes mellitus is on the rise due to overeating and lack of exercise. It is estimated that about 3.5 million people in 2010 are diabetic. In Korea, insulin-independent diabetes mellitus is now a serious problem in economic and social health. According to the statistics for 2013, diabetes mellitus falls within the top 10 of the elderly people aged 65 and older, and the annual reported healthcare costs reach 219.28 billion won. Diabetes treatment drugs are mainly used for blood sugar control and delayed complications. Oral glucose-lowering agents such as insulin preparations, sulphonylureas, acetaminophen, and alpha-glucosidase inhibitors are used. There are problems such as limitation of treatment. In addition, according to recent studies, it has been suggested that the intake of antioxidant active substances capable of removing active oxygen is becoming important for effective diabetes management because it causes various diabetic complications due to damage of specific cells due to active oxygen. However, most of the diabetic patients have difficulty in practicing even though they are aware of the purpose of dietary therapy. Therefore, there is a high preference for anti - diabetic foods which can be easily ingested. Although the development of products for functional sugar control is progressing actively, it focuses on the single efficacy, despite the diseases caused by various mechanisms.

In this regard, probiotic is a living microorganism that has a health and beneficial effect on the host when an adequate amount is ingested. The probiotic intake helps stabilize intestinal flora, maintains balance among intestinal bacteria, It is also known to help maintain. Probiotics contribute to the immune response, which increases the macrophage activity and the anti-inflammatory cytokine transcriptional regulatory factors by the cell wall components of the probiotic strains, resulting in an immune response to remove foreign substances such as infectious harmful microorganisms Because. However, there have been no reports on strains of plant - derived multi - type probiotics which have an inhibitory activity against dietary carbohydrate digestive enzymes and simultaneously possess antioxidative and probiotic activities.

Therefore, the present inventor has tried to find strains which can be used as probiotics from fermented foods and can solve the problems in products for preventing and / or improving diabetes. As a result, it has been found that, even without the inhibitory effect on alpha amylase, The present invention has been accomplished by confirming a strain which has an excellent inhibitory effect on sidase, shows an antioxidative effect and can be used as a probiotic.

Choi Hwang-yong and others 2012. Lim et al, 2011, Seo and Lee, 2007

The present invention is based on the finding that the leucono-stokemeteroid MKSR strain deposited with accession number KCTC18665P has a high survival rate against a high concentration of artificial intestinal fluid and can function as a probiotic, It has excellent inhibitory activity against alpha glucosidase, an enzyme that converts disaccharide maltose into glucose and has excellent antioxidative effect, antimicrobial activity against food poisoning bacteria, And diabetes prevention and improvement effects.

In addition, the health functional food containing the above-mentioned strain, culture solution thereof or a mixture thereof as an active ingredient provides excellent diabetes prevention and improvement effect, antibacterial effect and antioxidative effect.

Further, a food additive containing the strain, the culture solution thereof, or a mixture thereof as an active ingredient provides an excellent effect of producing a complex carbohydrate or an oligosaccharide.

The present invention provides a Leuconostoc mesenteroides MKSR strain deposited with accession number KCTC18665P.

The present invention also provides a health functional food for preventing or ameliorating diabetes, comprising as an active ingredient, a culture strain of Leuconostoc mesenteroides MKSR (MKSR) deposited with deposit number KCTC18665P, a culture thereof or a mixture thereof.

The present invention also provides a food additive containing as an active ingredient, a culture of Leuconostoc mesenteroides MKSR ( Leuconostoc mesenteroides MKSR) deposited with the deposit number KCTC18665P, a culture thereof or a mixture thereof.

The present invention is based on the finding that the leukoinostomenseoride MKSR strain deposited with accession number KCTC18665P has a high survival rate with respect to a high concentration of artificial intestinal fluid and can function as a probiotic and has a side effect The present invention has no inhibitory effect of alpha amylase in solving abdominal fullness and has an excellent inhibitory activity against alpha glucosidase, an enzyme that converts disaccharide maltose to glucose, and has excellent antioxidative activity, antimicrobial activity against food poisoning bacteria, Thereby preventing and improving diabetes mellitus.

In addition, the health functional food containing the above-mentioned strain, culture solution thereof or a mixture thereof as an active ingredient provides excellent diabetes prevention and improvement effect, antibacterial effect and antioxidative effect.

Further, a food additive containing the strain, the culture solution thereof, or a mixture thereof as an active ingredient provides an excellent effect of producing a complex carbohydrate or an oligosaccharide.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a diagram of the morphology of leucono-stokemeteroide MKSR.
Fig. 2 relates to SEQ ID NO: 1 as a result of 16s rRNA analysis of leucono stokemeteroid MKSR. 16s rRNA identification result Leuconostoc mesenteroides strain ATCC 8293.
FIG. 3 is a graph mesenteroides MKSR (phylogenetic tree).
Figures 4 and 5 illustrate different Leuconostoc the ability to utilize sugars according to the sugars added to the medium of the mesenteroides species, whether oligosaccharide and dextran are produced. Leuconostoc M, 1% maltose on the production of oligosaccharides and dextran using different carbohydrates through fermentation of mesenteroides MKSR; G, 1% glucose, S, 1% Sucrose, F, 1% Fructose; O, 1% Oligosaccharide; LMG, LM medium + 1% glucose; LMM, LM medium + 1% Maltose; LMS, LM medium + 5% Sucrose; LMF, LM medium + 1% Fructose; LMGS, LM medium + 1% Glucose + 5% Sucrose; LMMS, LM medium + 1% Maltose + 5% Sucrose; LMFS, LM medium + 1% Fructose + 5% sucrose.
Figure 6 is a graphical representation of Leuconostoc mesenteroides MKSR inhibitory effect of alpha glucosidase.
FIG. 7 is a photograph related to a hemolysis reaction test result, wherein (A) shows Leuconostoc mesenteroides MKSR, (B) shows Leuconostoc mesenteroides subsp. mesenteroides (hereinafter LM) (C) are Leuconostoc citreum (hereinafter referred to as LC) (D) was developed for Staphylococcus aureus .

The present invention provides a Leuconostoc mesenteroides MKSR strain deposited with accession number KCTC18665P. The strain is a probiotic strain having an alpha-glucosidase inhibitory activity, and it has no inhibitory effect on alpha amylase, and it can improve the problem of abdominal bloating which is a side effect of acabose, which is used as a conventional diabetes medicine. Also, it can be used as a leuko-stokemesteroid MKSR probiotic deposited with the deposit number KCTC18665P, and has an antioxidative effect. In addition, the strain is characterized by having an antibacterial effect against food poisoning bacteria. Probiotics enable the β-glucan of lactic acid bacteria to regulate and inhibit cholesterol by structural and chemical properties, thereby lowering cholesterol absorption. Therefore, when probiotics are consumed for a long period of time, they may have an effect of preventing arteriosclerosis or hyperlipemia. Examples of the strains usable as probiotics include Leuconostoc genus, Enterococcus genus, Streptococcus genus Bacillus , and the like. Leuconostoc genus is more preferably used in the present invention. In relation to the function of the probiotics strain, various effects such as survival ability of digestive juice, antimicrobial effect against pathogenic bacteria, strengthening of immune function, normalization of cholesterol level and improvement of intestinal health can be demonstrated. In addition, it can also improve the whitening by ingesting probiotics. Probiotics can inhibit active oxygen production and have a high antioxidant activity, which can help improve whitening through inhibition of melanocyte formation. Therefore, the strain of the present invention may be a probiotic strain for the prevention or improvement of diabetes.

The present invention also provides a health functional food for preventing or ameliorating diabetes, comprising as an active ingredient, a culture strain of Leuconostoc mesenteroides MKSR (MKSR) deposited with deposit number KCTC18665P, a culture thereof or a mixture thereof. The strain has no alpha-amylase inhibitory activity and has alpha-glucosidase inhibitory activity, thereby improving the problem of abdominal bloating, which is a side effect of acabose, a conventional diabetes treatment agent. Also, since it can be used as a probiotics, it is possible to improve side effects and therapeutic limitations of organs such as alpha-glucosidase inhibitor for the treatment of diabetes, and to prevent diabetes and / An improvement effect can be provided. In addition, the strain of the present invention has an antioxidant effect, thereby mitigating diabetic complications due to cellular damage caused by reactive oxygen species, thereby contributing to the prevention and / or improvement of diabetes.

The present invention also provides a food additive containing as an active ingredient, a culture of Leuconostoc mesenteroides MKSR ( Leuconostoc mesenteroides MKSR) deposited with the deposit number KCTC18665P, a culture thereof or a mixture thereof. The strain can produce at least one selected from the group consisting of mannitol, fructose, oligosaccharide and dextran, and has an antimicrobial activity against food poisoning bacteria and the like, so that it can exert its role as a food additive. In addition, the strain can be used as a probiotics having an antioxidative effect, and a whitening effect can also be obtained.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in more detail with reference to examples and drawings of the present invention, but the scope of the present invention is not limited by the following embodiments.

Example  One. Leuconostoc mesenteroides MKSR  Specification of microbiological characteristics

Leuconostoc mesenteroides MKSR were cultured using MRS medium at 25-40 째 C, more preferably 25-38 째 C. It is possible to cultivate the microorganism in the temperature range described above, and it is difficult to cultivate the microorganism when the temperature is out of the above range. Microscopic observations, Gram staining, endospore formation and catalase activity were measured for the microbiological characteristics of MKSR. API 50CHL and API zym (API bioMerieux, France) were used to investigate carbohydrate availability and enzyme production ability. In addition, the nucleotide sequence of the 16s rRNA of MKSR was assigned to Solgent (Daejeon, Korea). One colony cultured was diluted and genomic DNA was extracted using a genomic DNA prep kit. The genomic DNA of the bacteria was amplified using the forward primer 27F (5'-AGA GTT TGA TCC TGG CTC AB-3 ') and the reverse primer 1492R (5'-GGT TAC CTT GTT ACG ACT T-3' PCR reaction mixture was prepared by mixing 1 μl of template, 10 μl of 2X Taq PCR premix (Solgent), 2 μl of forward primer (10 pmole / μl), and 5 μl of distilled water, and PCR (Biosystems, Life Technologies, Marsiling, Singapore ) Were performed. The PCR reaction was carried out at 96 ° C for 10 seconds, at 50 ° C for 5 seconds, at 60 ° C for 4 minutes, and repeated 30 times in total. After the PCR was completed, the PCR product was cleanly purified using the Solgent PCR purification kit and the PCR results were confirmed using 1% agarose gel. The purified DNA was sequenced using ABI 3730XL DNA analyzer (Applied Biosystems, Foster, USA) and then analyzed using BLAST (Basic Local Alignment Search Tool). The phylogenetic tree was constructed by selecting the species closely related to the 16s rRNA and MKSR of the strain by the MEGA7 program.

As a result, Leuconostoc The mesenteroides MKSR (hereinafter referred to as MKSR) did not form sporozoites with gram positive bacilli, and catalase activity was negative (see FIG. 1).

Example  2. Leuconostoc mesenteroides MKSR The party  reaction

MKSR is 0.5% (w / v) yeast extract, 0.5% (w / v) peptone, 2% (w / v) K 2 HPO 4 0.02% (w / v) MgSO 4 · 7H 2 O, 0.001% (w / v) for NaCl, the medium consisting of 0.001% (w / v) FeSO 4 · 7H 2 O, 0.001% (w / v) MnSO 4 · H 2 O, 0.013% (w / v) CaCl 2 · 2H 2 O In addition, different sugars such as 1% Maltose, 1% Fructose, 1% Glucose, 5% Sucrose, 1% Maltose and 5% Sucrose, 1% Fructose and 5% Sucrose, 1% Glucose and 5% The cells were incubated for 18 hours, centrifuged at 10,000 rpm for 5 minutes, and reacted with each other. The reaction products were muched on TLC silica gel 60 (EMD Millipore Co., USA) plates, developed and developed.

The ability of the sugar to be added to the medium to which the MKSR, LM, LC was added and oligosaccharide and Dextran production were examined (see FIGS. 4 and 5 and Table 1). Dextran, produced by lactic acid bacteria involved in kimchi fermentation, a traditional fermented food, has been approved for use as a food additive. Among the polysaccharides produced by lactic acid bacteria outside the cells, glucose, which is a constituent sugar, is a polysaccharide mainly linked by α- (1 → 6) bond and is actively used in the food industry such as prevention of crystals of sucrose products and control of physical properties of fluid foods. Representative Leuconostoc producing Dextran mesenteroides is a lactic acid fermentation strain and is a lactic acid bacterium that produces dextran using sucrose which is a fermenting sugar as a substrate. MKSR cultured on maltose supplemented media produced dextran with oligosaccharide. The Leuconostoc strain was reported to secrete dextransucrase as an extracellular enzyme, biosynthesizing polymer dextran as a polymer of glucose, and having physicochemical properties such as stickiness, hygroscopicity and thermal stability, and plays an important role in controlling the properties of food. In addition, Leuconostoc lactic acid bacteria produce sugars such as mannitol and fructose, which are sugar alcohols, and play a role in imparting sweetness and refreshing taste to fermented products. Sugar alcohols produced by lactic acid bacteria have advantages over the production method by hydrogenation at high temperature and high pressure under a high temperature and chemically, and many studies have been conducted on the production of mannitol by microorganisms.

Table 1 below shows the effect of different carbohydrates on Leuconostoc mesenteroides And the results of the oligosaccharide produced by the fermentation of MKSR and whether or not dextran is formed are shown.

[Table 1]

^* M, 1% maltose; G, 1% glucose, S, 1% sucrose; F, 1% fructose; O, 1% oligosaccharide; LMG, LM medium + 1% glucose; LMM, LM medium + 1% maltose; LMS, LM medium + 5% sucrose; LMF, LM medium + 1% fructose; LMGS, LM medium + 1% glucose + 5% sucrose; LMMS, LM medium + 1% maltose + 5% sucrose; LMFS, LM medium + 1% fructose + 5% sucrose

Table 2 below shows the results of an experiment on the carbohydrate fermentation ability of MKSR. MKSR is a L. mesenteroides sp . mesenteroides (LM), and there was a difference in glucose utilization such as Xylose, D-lactose and Potassium gluconate.

[Table 2]

¹ MKSR, Leuconostoc mesenteroides MKSR; LM, Leuconostoc mesenteroides ; LC, Leuconostoc citreum

² +, positive reaction; -, negative reaction

Example  3. Leuconostoc mesenteroides MKSR Acid resistance and My bile  And viability

After incubation at 30 ° C for 18 hours, the cells were inoculated into 0.1% pepsin solution (pH 2.0; Sigma-Aldrich Co.) and cultured at 37 ° C for 3 hours. Was inoculated on 3% bile salt (pH 6 ~ 7; Sigma-Aldrich Co.), and the cells were plated on MRS agar for each hour and cultured in a 30 ° C incubator for 24 hours.

Lactic acid bacteria, as a probiotic, are required to survive in an environment in which gastric acid and bile are present in order to exert a formal function and various physiological functions in the human body. Cholanic acid inhibits the growth of microorganisms by affecting the cell membrane of lipid-containing microorganisms. In many species of lactic acid bacteria including Lactobacillus , hydrolysis of bile acids and reduction of bile acid hydrolysis enzymes are inhibited (Lee, Kyunghee, et al. , 2016). To measure the tolerance to acid and biliary properties of MKSR, strains were inoculated and viable counts were measured over time (see Table 3). Table 3 below shows that Leuconostoc mesenteroides The number of MKSRs survived when the MKSR was passed through the conditions of pepsin (pH 2) and 3% bile salt solution is described. MKSR is likely to be available as a probiotic for strains that survive in the intestinal environment and survive 100% when passed for 3 hours in 0.1% pepsin solution and 4 hours in 3% bile salt solution.

Table 3 below shows that when pepsin (pH 2) and 3% bile acid are passed, the viable euconostoc mesenteroides This is a table regarding the number of MKSR cells.

[Table 3]

Example  4. Leuconostoc mesenteroides MKSR  Antimicrobial activity

The supernatant is collected by centrifugation at 10000 rpm for 5 minutes at the culture broth cultured in the MRS liquid medium for 18 hours. Some of the supernatant is adjusted to pH 5.5-6.5 with 1N NaOH and some pH is not adjusted. The pH-adjusted supernatant and the untreated supernatant were filtered through a 0.2 μm filter. (BHI, MB cell Co., Korea), TSA (Tryptic Soy agar, Difco Co., USA) and NA (Nutrient Agar, Difco Co., USA) ). After discs were spread, 20 μl of the bacterial supernatant was inoculated and the size was measured in mm in which bacteria did not grow around the disc at 14 hours, 24 hours, and 48 hours.

The antimicrobial activity of MKSR was determined by the presence of gram-negative bacteria such as E. coli , Pseudomonas aeruginosa , Shigella sonnei, S. flexneri , Klebsiella pneumonia and gram-positive bacteria such as Bacillus cereus, Listeria monocytogenes , L. innocua , Staphylococcus aureus And then exposed for 14, 24, and 48 hours, respectively. As a result, MKSR showed 14 hours of Pseudomonas aeruginosa , Shigella sonnei , S. flexneri , Bacillus cereus , Listeria monocytogenes , L. innocua, Staphylococcus aureus , 24 hours Pseudomonas aeruginosa , Shigella sonnei, S. flexneri , Bacillus cereus , L. innocua When cultured for 48 hours, Pseudomonas aeruginosa , Shigella sonnei , S. flexneri , Klebsiella pneumonia , Bacillus cereus , and L. innocua (Table 6). In a study on the antimicrobial activity of Lactobacillus sp. Isolates from shrimp farms, it was reported that the antimicrobial activity by organic acids produced during culture and the neutralization of culture supernatant did not show antimicrobial activity (Ma et al 2009), Choi et al. (2014) also reported that antimicrobial activity was not observed in the culture supernatant with pH neutral control, but the antimicrobial activity was observed in the supernatant without pH control. MKSR had antimicrobial activity against some strains when the pH was adjusted to neutral (see Table 4). The detailed description of the inhibitory zone against the pathogenic bacteria of the present invention Leuconostoc mesenteroides MKSR is shown in Table 4 below.

[Table 4]

(Unit: mm)

Example  5. Leuconostoc mesenteroides MKSR  Antioxidant activity

DPPH radical scavenging activity

100 μl of cell free supernatant or intact Cell was added to 1.2 ml of 0.2 mM DPPH (1-1 diphenyl-2-picrylhydrazyl) (Sigma-Aldrich Co.). The absorbance at 517 nm was measured using a spectrophotometer (Cary 60 UV-Vis, Agilent Technologies Inc, Santa Clara, CA, USA). (zero methanol)

Ferric reducing antioxidant power (FRAP) -

FRAP reagent [10 mM TPTZ (2,4,6-tripyridyl triazine) (Sigma-Aldrich Co.): 20 mM FeCl ₃ .6H ₂ O: 0.3 M sodium acetate buffer (1: 1: After preheating at 37 ° C, 50 μl of Cell free supernatant or Intact Cell was added and reacted in the dark for 30 min. Absorbance was measured at 593 nm using a spectrophotometer to determine the color change. FeSO ₄ · 7H ₂ O was used as a standard sample.

SOD-like activity

1.3 ml of Tris-HCl buffer (50 mM tris [hydrowymethyl] amino-methane + 10 mM EDTA) (pH 8.0) was added to 0.1 ml of a cell-free supernatant or Intact Cell, and 0.1 ml of 7.2 mM pyrogallol (Sigma Aldrich Co.) Lt; / RTI > The reaction was stopped by adding 50 1 of 1N HCl, and the absorbance was measured at 420 nm using a spectrophotometer.

After experiments according to the above, Leuconostoc Specific results for the antioxidant activity of mesenteroides MKSR are shown in Table 5.

Recent research suggests that certain cell damage caused by active oxygen causes various diabetic complications, thus emphasizing the importance of taking antioxidant substances that can remove active oxygen for effective diabetes management (Dembinska-Kiec et al. 2008). DPPH has a free radical, and when it reacts with a substance having antioxidative activity, the radical is extinguished with giving off electrons, its specific purple color becomes transparent and yellowish, and the ability to reduce radical, that is, the radical scavenging activity, Is high. The DPPH radical scavenging effect was measured and the MKSR scavenging effect was high. In the study of Lee et al. (2016), Lactobacillus rhamnosus GG (7 log CFU / ml) was about 64% lower than that of MKSR. MKSR is also described by Chen et al. (8 ~ 30%) of the 12 strains studied in this study (2014).

Reducing power is an important factor related to antioxidant ability. FRAP assay is a method of measuring the ability to reduce the Fe ^{3 +} -ferricyanide complex to a Fe ^{2 +} form by reducing it to blue (Jeon et al. 2013). The reducing power of MKSR was 3.36-3.44 FeSO _{4 ·} 7H ₂ O eq mM, and the reducing power of 1 mg / ml ascorbic acid used as a control was 4.06 FeSO _{4 ·} 7H ₂ O eq mM.

In addition to the radical elimination of free radicals by converting active oxygen present in the body into hydrogen peroxide, H ₂ O ₂ produced by SOD is converted to water molecules and oxygen molecules by peroxidase or catalase The enzyme plays an important role in protecting the body. Similar activity of MKSR was observed only in the cell free supernatant (see Table 5). Table 5 below shows the antioxidative activity of Leuconostoc mesenteroides MKSR.

[Table 5]

Example  6. Leuconostoc mesenteroides MKSR  Carbohydrate digestive enzyme inhibitory activity

α-glucosidase inhibition -

(Dextrose 2% (w / v), Peptone 1% (w / v), Beef (w / v)) or the like were added to 50 μl of 0.02 M sodium phosphate buffer (pH 6.8) (w / v), 0.5% (w / v) of yeast, 0.1% (w / v) of Polysorbate 80, 0.2% 50 μl of the Intact Cell was cultured in a medium prepared by adding 0.01% (w / v), Manganese phosphate 0.005% (w / v), and Dipotassium phospate 0.2% (w / v) After adding 200 μl of 2 mM pNPG (Sigma-Aldrich Co.), 50 μl of a-glucosidase (Sigma-Aldrich Co.) was added and reacted in a water bath at 37 ° C for 40 minutes. In order to stop the reaction, 1.5 ml of 0.1 M Na ₂ CO ₃ was added, and the absorbance at 405 nm was measured using a spectrophotometer. 0.5mM pNP was used as a standard sample.

α-amylase inhibition -

Add 50 μl of Cell free supernatant or Intact Cell to 50 μl of 0.02M sodium phosphate buffer (pH 6.8). After adding 1% starch, 50 μl of a-amylase (Sigma-Aldrich Co.) is added and reacted in a water bath at 37 ° C for 20 minutes. 250 μl of dinitrosalicylic acid (DNS) was added, reacted at 95 ° C for 5 minutes, and 2 ml of distilled water was added. The color change was measured at 540 nm using a spectrophotometer. 10 mg / ml maltose was used as a standard sample.

Table 6 shows the enzymatic activity of MKSR. MKSR showed acid phosphatase, Naphtaol-AS-BI-phosphohydrolase, α-galactosidase, β-galactosidase, α-glucosidase and β-glucosidase activity. In particular, α-galactosidase is an enzyme that degrades saccharides such as raffinose and stachyose, which are present in many cereals such as soybean and cause gas production in the large intestine, so that a dressing effect can be expected in humans and animals.

[Table 6]

Table 6 above relates to the number of viable MKSR cells when passed with pepsin (pH 2) and 3% bile acid solution. It is known that oral hypoglycemic agents, a method of treating type 2 diabetes, use substances that inhibit enzymes involved in glucose degradation. Among the enzymes involved in the degradation of sugars, α-glucosidase and α-amylase play an important role. Disaccharides as monosaccharides and the latter as polysaccharides into small sugars. These inhibitors of sugar enzymes inhibit the degradation of sugars and ultimately inhibit the absorption of complex carbohydrates, so glucose is absorbed into the entire small intestine, which is advantageous in that postprandial increase in blood glucose levels is gentle. Currently, acarbose is used as an inhibitor of sugar enzymes. However, acarbose, which has a high inhibitory activity against both α-glucosidase and α-amylase, may have stomach-related symptoms such as abdominal distension and diarrhea due to side effects.

The α-glucosidase inhibition of MKSR was high, but there was no α-amylase inhibitory activity (Table 7). In the same experimental conditions, α-glucosidase inhibition of acarbose was about 111%, which was very high, and there was no α-amylase inhibitory activity. Chen et al. (2014) reported that the intact cell and cell-free supernatant of Lactobacillus rhamnosus GG (LGG), one of the probiotic, inhibited α-glucosidase activity by 11.4% and 29.57%, respectively. Although α-glucosidase inhibition of LGG was lower than that of MKSR, inhibition of α-amylase was also observed. The above results suggest that MKSR is a good alternative to acarbose's side effects.

[Table 7]

Table 7 shows that Leuconostoc mesenteroides MKSR inhibitory activity of α-glucosidase and α-amylase (Unit:%)

The following Table 8 relates to a media composition of MKSR for the production of inhibitors for [alpha] -Glucosidase.

[Table 8]

(Unit: g / L)

α-Glucosidase inhibitory activity was higher when cultured in medium # 1, # 2, # 5, # 7, # 9, and # 4 (see FIG. 6). According to Brenda (BRaunschweig Enzyme Database), inhibitor of α-glucosidase it is to say such as Mg ^{2 +,} Mn ^{2 +.} However, the Fig. # 7, 6, a group without addition of Mg ^{2 +,} Mn ^{2 +} In the # 8 Mg ^{2 +,} Mn ^{2 +} in the medium composition is not added when viewed as having α-glucosidase inhibitory activity, MKSR bacteria itself α-glucosidase inhibitory activity.

Example  7. Leuconostoc mesenteroides MKSR  Hemolytic test

The cells were incubated at 37 ° C for 4 h at 37 ° C in a Tryptic soy + 5% sheep blood (Kisan Bio Co., Korea) solid medium. Staphylococcus aureus was used as a positive control.

Hemolysis is an abnormal action that is caused by the normal action of erythrocytes destruction, genetic defects of erythrocytes, toxic substances such as chemicals, snakes, and microorganisms. When haemolysis occurs in cells, oxygen transport function of red blood cells is lost, resulting in fatal results in vivo (Yoon, HJ et al., 2013). To confirm the safety of the selected lactic acid bacteria, the presence or absence of hemolysis was evaluated and the results as shown in Figure 6 were obtained. You can see that handsome ring of red blood cells are destroyed around the transparent cells in the control of S. aureus, and red blood cells are destroyed around the cells in all three types of lactic acid bacteria used in this study hemolytic test was confirmed that the transparent ring does not occur Safety ( 7).

Korea Biotechnology Research Institute KCTC18665P 20180126

Claims (10)

Leuconostoc mesenteroides MKSR strain deposited with accession number KCTC18665P. The strain according to claim 1, wherein the strain is a probiotic strain having an alpha glucosidase inhibitory activity. The strain according to claim 1, wherein the strain is a probiotic strain having no inhibitory action of alpha amylase. The strain according to claim 1, wherein the strain is a probiotic strain for diabetes prevention or improvement. The strain according to claim 1, wherein the strain is a probiotic strain having an antioxidative activity. A health functional food for preventing or ameliorating diabetes, comprising as an active ingredient, a culture of Leuconostoc mesenteroides MKSR (MKS) deposited with the deposit number KCTC18665P, a culture thereof or a mixture thereof. The dietary supplement according to claim 6, wherein the strain is a probiotic strain having an alpha glucosidase inhibitory activity. The health functional food for preventing or ameliorating diabetes according to claim 6, wherein the strain is a probiotic strain having no inhibitory action of alpha amylase. A food additive containing a strain of Leuconostoc mesenteroides MKSR (MKSR) deposited with the deposit number KCTC18665P, a culture thereof or a mixture thereof as an active ingredient. The food additive of claim 9, wherein the strain produces at least one selected from the group consisting of mannitol, fructose, oligosaccharide and dextran.

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