KR101234582B1 - Composition for α-glucosidase inhibitory activity - Google Patents

Abstract

The present invention relates to a composition for inhibiting alpha glucosidase activity, and more particularly, to a composition containing a Pediococcus spissis JNU534 strain or a culture thereof having alpha glucosidase inhibitory activity. Since the composition is excellent in inhibiting the activity of acid and bile resistance and alpha-glucosidase, when included in dairy products, health foods and food additives, there is no side effect, and the amount of low sugar absorbed into the body is fundamentally reduced. Not only can the effect be obtained, but also has the advantage that it can be usefully used in the preparation of anti-diabetic prevention or treatment, including high value-added hypoglycemic agents.

Description

Composition for inhibiting alpha glucosidase activity {Composition for α-glucosidase inhibitory activity}

The present invention relates to a composition for inhibiting alpha glucosidase activity, and more particularly, to a composition containing a pediococcus spissis JNU534 strain or a culture thereof having alpha glucosidase inhibitory activity.

Diabetes is a disease caused by the body's inadequate use of carbohydrates in food, resulting in too high a concentration of glucose in the blood. The hypoglycemic agent used to treat diabetes is to control postprandial blood sugar and not cause hypoglycemia. In the case of administration of insulin preparations, sulfonylurea preparations, and biguanide preparations, which are commonly used as oral hypoglycemic agents, blood glucose levels increase rapidly after eating, while insulin blood My appearance is relatively slow, so postprandial hyperglycemia, hyperinsulinemia and hypoglycemia between meals.

Therefore, in order to overcome the problems of the conventional method of controlling the glucose concentration in the blood, a new method of preventing the absorption of glucose in the blood itself has been recently attempted. One of the new methods has been attempted to develop alpha-glucosidase inhibitors as a diabetes treatment. Since the alpha-glucosidase inhibitor inhibits glucose absorption in the blood itself, the alpha-glucosidase inhibitor may be used for the treatment and prevention of obesity and hyperlipidemia in addition to diabetes, and the representative therapeutic agents include acarbose (Acarbose, Glucobay, Bayer). .

Meanwhile, more than 100 trillion intestinal bacteria live in our bodies. If the intestinal bacteria have a large force, they can maintain their health by defeating the disease-causing bacteria. However, if the forces of harmful bacteria are large, the immunity is lowered, helping the activity of pathogens, causing various diseases such as stomach cancer and colon cancer. Lactic acid bacteria, which is the best among the beneficial bacteria, are widely distributed in the natural world such as human or mammal's digestive tract, oral cavity, vagina, various fermented foods and soils. Able to know.

The action of the lactic acid bacterium enhances the immune function of the host, inhibits the absorption of cholesterol in the intestine, contributes to the prevention of adult diseases, inactivates the carcinogens in the intestine, and inhibits the enzyme activity involved in carcinogenesis. It is known that there is an effect of suppressing the occurrence of the back. In addition, it acts in various ranges such as inhibiting intestinal harmful bacteria and intestinal action, anti-aging, skin cosmetic effect, lactose intolerance, mutation suppression effect.

As described above, attempts have been actively made to develop lactic acid bacteria having excellent physiological efficacy as functional foods or therapeutic agents, and the present applicant has also applied for Pediococcus spissis JNU534 in Korean Patent Registration No. 10-0890462. However, it is not disclosed that the Pediococcus spissis JNU534 strain has a hypoglycemic effect by inhibiting the action of alpha-glucosidase.

Accordingly, the present inventors, while overcoming the problems of the side effects of the conventional therapeutics, such as microorganisms, herbal medicines, food ingredients to search for an activity inhibitory agent of alpha-glucosidase, by the present applicant The activity of inhibiting activity against alpha-glucosidase of the Pediococcus spissis JNU534 strain (Accession No. KCCM-10869P) disclosed in the first issue was first discovered and the present invention was completed.

It is an object of the present invention to provide a composition for inhibiting alpha glucosidase activity, comprising a Pediococcus spissis JNU534 strain or a culture thereof.

It is yet another object of the present invention to provide pharmaceuticals, dairy products, health functional foods and food additives which contain the composition and which can reduce the amount of low saccharides absorbed into the body by inhibiting the activity of alpha glucosidase. will be.

In order to achieve the above object, the present invention provides a composition for inhibiting alpha glucosidase activity, comprising a Pediococcus spissis JNU534 strain or a culture thereof.

The present invention provides pharmaceuticals, dairy products, dietary supplements and food additives that contain the composition and which can reduce the amount of low saccharides absorbed into the body by inhibiting the activity of alpha glucosidase.

Hereinafter, the present invention will be described in detail.

In the present invention, "culture" means that a specific microorganism is cultured in a culture medium or a culture medium, and the culture may or may not include the specific microorganism. The culture is not limited in its formulation, and may be, for example, liquid or solid.

As used herein, the term "composition" is a concept that includes not only conventional foods, functional foods or dietary supplements, but also those approved as pharmaceuticals.

The present invention comprises a culture of Pediococcus sp. JNU534 (Accession No. KCCM-10869P) strain or Pediococcus sp. JNU534 (Accession No. KCCM-10869P) strain It provides a composition for inhibiting alpha glucosidase activity.

More specifically, Pediococcus sp. (Pdiococcus sp.) JNU534 (Accession No. KCCM-10869P) strain of the present invention can be isolated from kimchi, and inhibits the growth of harmful microorganisms or microorganisms that cause food decay. Have.

Inhibition of the activity of alpha-glucosidase was investigated in various lactic acid bacteria including the strain. As a result, we found for the first time the activity of Pediococcus spissis JNU534, which is lower than acarbose which shows 98% of inhibitory activity as a control, but the highest inhibitory activity among the lactic acid bacteria used in the experiment. Was done. This is a result of confirming that the pediococcus spissis JNU534 strain according to the present invention has a hypoglycemic effect by inhibiting the action of alpha-glucosidase.

The present invention provides a health functional food, a food additive, and a fermented dairy product containing the composition for inhibiting alpha glucosidase activity.

The composition includes not only health functional foods, but also general foods that humans consume in general and daily. When used as a nutraceutical, it may be formulated into a formulation of conventional nutraceuticals known in the art, together with a pharmaceutically acceptable carrier or additive. The health functional food may be prepared, for example, as a powder, granules, tablets, capsules, suspensions, emulsions, syrups, liquids, extracts, teas, jellies, beverages, and the like. As a food acceptable carrier or additive, any carrier or additive known to be available in the art for the preparation of the formulation to be prepared may be used.

Preparation of the health functional foods, food additives and fermented dairy products containing the Pediococcus spissis JNU534 strain according to the present invention and their contents can be easily determined from conventional methods known in the art, It may also contain other substances for maintaining the lactic acid bacteria viability.

In the case of taking the composition of the present invention periodically, the microorganism of the present invention in the intestine forms a bacterium and inhibits the activity of alpha glucosidase, thereby reducing the amount of low saccharides absorbed into the body, resulting in the composition of the present invention. This prevents and treats diabetes.

In addition, Pediococcus spissis JNU534 strain according to the present invention can be used in the preparation for the prevention or treatment of anti-diabetic. In such cases, they may be formulated in conventional pharmaceutical formulations known in the art. The medicament is preferably formulated into oral formulations and may be formulated into oral dosage forms such as, for example, solutions, suspensions, powders, granules, tablets, capsules, pills or expectorates.

When formulated into each of the above formulations, it may be prepared by the addition of a pharmaceutically acceptable carrier or additive necessary for the preparation of each formulation. Representatively, when formulated into a dosage form for oral administration, one or more of a diluent, a lubricant, a binder, a disintegrant, a sweetener, a stabilizer, and a preservative may be used as the carrier, and as an additive, flavors, vitamins, and antioxidants may be used. One or more types can be selected and used out of them.

The carrier and the additive may be any pharmaceutically acceptable, and specifically, lactose, corn starch, soybean oil, microcrystalline cellulose, or mannitol as a diluent, magnesium stearate or talc as a lubricant, and polyvinylpyrrolidone as a binder. Or hydroxypropyl cellulose is preferred. In addition, as a disintegrant, carboxymethyl cellulose calcium, sodium starch glycolate, potassium polyacrylic acid, or crospovidone, sweetener as white sugar, fructose, sorbitol, or aspartame, stabilizer as carboxymethyl cellulose sodium, beta-cyclodextrin, As lead, or xanthan gum, and preservative, methyl paraoxybenzoate, propyl paraoxybenzoate, or potassium sorbate is preferable.

In addition to the above components, in order to enhance the taste as a known additive, natural flavors such as plum flavor, lemon flavor, pineapple flavor, herbal flavor, natural pigments such as natural fruit juice, chlorophyllin, flavonoid, fructose, honey, sugar alcohol, sugar Sweetening ingredients such as, or may be used by mixing an acidulant such as citric acid, sodium citrate.

Such formulation methods and carriers and additives required for formulation are described in detail in Remington ' s Pharmaceutical Sciences (19th ed., 1995).

The composition for inhibiting alpha glucosidase activity according to the present invention is excellent in inhibiting the activity of acid, bile and alpha-glucosidase, and when included in dairy products, health functional foods and food additives, there are no side effects and The effect of reducing the amount of low saccharides absorbed can be obtained at source.

The composition for inhibiting alpha glucosidase activity according to the present invention has an advantage that it can be usefully used for the preparation of an anti-diabetic prophylactic or therapeutic agent including a high value-added hypoglycemic agent as an inhibitory activity of the excellent alpha glucosidase.

Hereinafter, the present invention will be described in more detail with reference to specific examples. However, the present invention is not limited by the following examples, and it is apparent to those skilled in the art that various changes or modifications can be made within the idea and scope of the present invention.

Here, unless otherwise defined in the technical terms and scientific terms used, those having ordinary skill in the art to which the present invention belongs have a generally understood meaning.

Repeated descriptions of the same technical constitution and operation as those of the conventional art will be omitted.

[ Example  1] alpha Glucosidase (α- glucosidase ) Inhibition activity measurement

Pediococcus spissis JNU534 strain (Accession No. KCCM-10869P) of the present embodiment is pediococcus spissis JNU534 strain (deposited institution: Korea microbial preservation affiliated with the Korean spawn association) in the Republic of Korea Patent No. 10-0890462 Center, Accession No .: KCCM-10869P, Accession Date: May 2, 2007), the following experiment using the strain to inhibit the activity of alpha-glucosidase (α-glucosidase) The method was investigated.

20 µl of coenzyme solution and 50 µl of 6 mM p-nitrophenyl α-D-glucopyranoside (PNPG) solution were added to 60 µl of 20 mM Sodium phosphate (pH7.0) buffer, followed by reaction at 37 ° C for 30 minutes. Thereafter, 100 µl of 1M Glycine buffer (pH 9.0) was added, and then the absorbance was measured at 405 nm. One unit of enzyme activity was defined as the amount of enzyme that degrades 1 μmole of substrate in one minute.

Alpha-glucosidase inhibitory activity was determined by 40 μl of 20 mM sodium phosphate (pH7.0) buffer, 20 μl of coenzyme solution, 20 μl of sample, and 50 μl of 6 mM p-nitrophenyl α-D-glucopyranoside (PNPG) solution. After addition, it was made to react at 37 degreeC for 30 minutes. Thereafter, 100 μl of 1 M Glycine buffer (pH 9.0) was added, and the absorbance was measured at 405 nm.

As a result, as can be seen in Table 1, the active pediococcus spissis JNU534 showed an inhibition rate of 41.2% against alpha-glucosidase, which was lower than that of acarbose, which shows 98% inhibitory activity. It was confirmed that the highest inhibitory activity among the lactic acid bacteria. Pediococcus pentosaceus KCCM 40464 ( Pediococcus pentosaceus KCCM 40464) was found to exhibit inhibitory activity of 31.5% and Lactobacillus helveticus 1042 (28.4%).

From the above results, it can be confirmed that the diococcus spissis JNU534 strain of the present invention is less active against alpha-glucosidase and inhibits the activity of alpha-glucosidase compared to other lactic acid bacteria. It is also confirmed that it can be used as a material for diabetes.

Example 2 Evaluation of Acid Resistance of Pediococcus spissis JNU534 Strain

Pediococcus spissis JNU534 strains must be alive and passed through the gastrointestinal tract to be used as health functional foods, food additives, fermented dairy products or therapeutic agents. The artificial gastric fluid for evaluating resistance to gastric acid was sterilized after adjusting the pH of MRS broth to 2.5, 3.0, and 3.5 using 1 N-HCl, and then adding pepsin to 1000 unit / ml. It was used as a measuring gastric juice.

Evaluation of acid resistance of the Pediococcus spissis JNU534 strain was carried out as follows. After incubation at 37 ° C. in MRS broth, 10 ml of the solution was taken and centrifuged at 4,000 g for 20 minutes to recover the cells. The recovered cells were washed twice with sterile distilled water and diluted with physiological saline (0.85% NaCl) so that the number of viable cells was 10 ⁸ / ml, and 100 µl of the cell suspension was inoculated to 10 ⁶ / ml in the gastric juice. . Samples were taken immediately after inoculation, after 1 hour and after 2 hours while maintaining in a 37 ° C constant temperature water bath, diluted in 0.05% L-cysteine-added phosphate buffer (pH 7.2, 0.2 M), and then placed on MRS agar medium. After inoculating a certain amount of diluting solution of the colonies grown by incubating for 48 hours at 37 ℃ was shown in Table 2 below.

As can be seen from the results in Table 2, as a result of evaluating the acid resistance of the Pediococcus spissis JNU534 strain, it was confirmed that the initial viable count was maintained even at a low pH, which is pediococcus spissis JNU534 high acid resistance It is also the result of confirming probiotic lactic acid bacteria.

[ Example  3] Bile acid  Resistance evaluation

The resistance to bile acids against the Pediococcus spissis JNU534 strain was investigated. The resistance of bile acids was recovered in the Pediococcus spissis JNU534 strain by the same method as the acid resistance test of Example 2, and then initially added to the MRS liquid medium containing 0.25%, 0.5%, 0.75%, and 1.0% of oxgall. Viable cell count was added to 10 ⁵ / mL. After incubating at 37 ° C. for 24 hours, the number of viable cells when bile was added was examined and shown in Table 3 below.

As can be seen in Table 3, it was confirmed that lactic acid bacteria isolated from kimchi, but also resistant to bile acids.

The above results confirm that pediococcus spissis JNU534 strain and its cultures can function through the gastrointestinal tract and live in the intestine even when manufacturing healthy functional foods, food additives, fermented dairy products or therapeutic preparations. It is also.

As described above, the Pediococcus spissis JNU534 strain of the present invention is excellent in inhibiting the activity of acid resistance, bile resistance and alpha-glucosidase, drugs, dairy products, health functional foods to prevent or treat anti-diabetic And when included in food additives there is no side effect and can achieve the effect of reducing the amount of low saccharides absorbed into the body at the source.

[Formulation example 1 of the preparation for the prevention or treatment of antidiabetic diabetes] Tablets

80 mg of Pediococcus spissis JNU534 strain, 200 mg of galactooligosaccharide, 60 mg of lactose and 140 mg of maltose were mixed, granulated with a fluidized bed dryer, and then 6 mg of sugar ester was added. It was tableted with a tableting machine. The final weight of the content is 600 mg.

[Health of functional food Formulation Example  2] Drinks

Pediococcus spissis JNU534 strain culture 80 mg, glucose 10 mg, citric acid 0.6 g and liquid oligosaccharide 25 g was mixed and 300 ml of purified water was added to each bottle 200 ml. After filling the bottle sterilized for 4 to 5 seconds at 130 ℃ to prepare a beverage.

Claims (6)

Alpha Glucocontaining Cultures of Pediococcus sp. JNU534 (Accession No. KCCM-10869P) Strains or Pediococcus sp. JNU534 (Accession No. KCCM-10869P) Strains A composition for the prevention or treatment of antidiabetic, having a sidase inhibitory activity. A health functional food containing the composition for inhibiting alpha glucosidase activity according to claim 1. The method of claim 2,
The food is a health functional food, characterized in that one unit dosage form selected from powders, granules, tablets, capsules and drinks. Food additive containing a composition for inhibiting alpha glucosidase activity according to claim 1. A fermented dairy product containing the composition for inhibiting alpha glucosidase activity according to claim 1. An antidiabetic prophylactic or therapeutic agent containing the composition for inhibiting alpha glucosidase activity according to claim 1 as an active ingredient.