KR101466869B1 - Fermented soybean containing bacillus methylotrophicus tpp 0025 enhanced functionality of anti-diabetes and method of thereof - Google Patents

Abstract

The present invention relates to a soybean fermented product having enhanced antidiabetic effect and a method for producing the fermented soybean product, and more particularly, to a soybean fermented soybean fermented with an antidiabetic effect, And a step of fermenting soybeans inoculated with the strain, and a soybean fermentation product produced by the method. The present invention provides a fermented soybean having an antidiabetic activity due to a strain having high alpha glucosidase inhibitory activity.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a soybean fermented product having enhanced antidiabetic effect including Bacillus methylotrophicus TPP 0025,

The present invention relates to a soybean fermented product having enhanced antidiabetic effect and a process for producing the same.

As economic development accelerates and the food culture becomes more westernized and simplified, the incidence of metabolic syndrome related to this is gradually increasing. One of the representative diseases is diabetes. According to the Korean Diabetes Epidemiology Survey conducted by the Korean Diabetes Association and the HIRA, 2.7 million people (7.7% of the total population) suffered from domestic diabetes in 2005, 4.55 million (8.97% of the total population) by 2020, It is estimated to reach 5.45 million (10.85% of the total population). In addition, a nationwide preventive management system for diabetes is required, including 10% of new patients every year. Diabetes mellitus is divided into type 1 diabetes and type 2 diabetes according to the etiology of the disease. Type 2 diabetes, which accounts for 90% of all diabetes, is caused by insulin action due to various factors in muscle, liver, It is caused when the beta cells do not secrete insulin enough to overcome insulin resistance and the blood glucose concentration continues to be high. If diabetes occurs, the sugar utilization rate is lowered and the sugar is lost in the urine. As a result, the sugar can not be used as an energy source and thus the energy can not be produced. As a result, the protein in the body is used as an energy source, Feel and lose weight. In addition, patients with diabetic retinopathy and diabetic retinopathy, which are complications of diabetes in 2007, will experience 60% or more of a year-on-year increase in complications, such as decreased renal function, arteriosclerosis, decreased visual acuity due to retinal hemorrhage, And 36%, respectively. As such, the incidence of diabetes itself is increasing, and the accompanying complication rate is high. Therefore, the cost of medical care (one-fifth of total health care costs for diabetes patients) . Nevertheless, it is not yet possible to develop a drug that can cure diabetes.

Since insulin or oral hypoglycemic agent is administered to prevent or treat the above-mentioned diabetes, since it is accompanied by an economic burden and a risk of side effects, recently, dietary and natural therapies using pharmacological substances such as natural plants This is being attempted. As the natural products reported to be effective for the treatment of diabetes, there have been reported the extracts of Cucurbitaceae (Korean Patent No. 464815), the phthalic acid derivatives (Korean Patent No. 457690) and the extracts of Hovenia dulcifera (Korean Patent No. 417287) , Polysaccharides isolated from Angelica gigas Nakai (International Publication No. 2001-60386), and ruminant bile (US Patent No. 6451355).

On the other hand, Chungkookjang is Korea's representative fermented food produced by short fermentation for 2-3 days using Bacillus subtilis in soybean, and it is known that it has immunity enhancement, anticancer activity and is effective for prevention and inhibition of metabolic diseases such as diabetes and hyperlipidemia . However, since these various physiological activities of chongkukjang exhibit different activities depending on the kinds of seeds used in production, fermentation should be carried out using seeds having excellent functionality in order to produce chungkukjang having high physiological activity.

Thus, the present inventors isolated a strain having high alpha glucosidase inhibitory activity from a conventional fermented food, and produced a soybean fermented product exhibiting an excellent effect on postprandial blood glucose control through the inhibition of alpha glucosidase using the strain, .

Thus, it is an object of the present invention to provide a method of isolating a strain having high alpha glucosidase inhibitory activity from a conventional fermented food and using the fermented soybean fermented product to produce a soybean fermented product.

In order to achieve the above object, there is provided Bacillus methylotrophicus TPP 0025 (KCCM11279P), which has an alpha glucosidase inhibitory activity in one embodiment of the present invention.

In another embodiment of the present invention, the step of inoculating soybeans with a strain of Bacillus subtilis grown; And fermenting soybeans inoculated with the strain. The present invention also provides a method for producing soybean fermented product having enhanced antidiabetic effect.

In the present invention, the strain is selected from the group consisting of Bacillus methylotrophicus TPP 0025, KCCM11279P). The strain may be inoculated with 0.05 to 5% by weight of the amount of soybean added, and the soybean inoculated with the strain may be fermented under conditions of 60 to 80% of humidity, Can be fermented at a temperature of 30 to 45 DEG C, and the soybean inoculated with the strain can be fermented for 1 to 5 days.

In another embodiment of the present invention, there is provided a food comprising the fermented soybean having an improved antidiabetic effect according to the above method as an active ingredient.

In another embodiment of the present invention, there is provided a soybean fermentation product enhanced in an antidiabetic effect comprising a strain of Bacillus subtilis and a soybean grown therein.

In the present invention, the strain may be Bacillus methylotrophicus TPP0025 (KCCM11279P), and the strain may be 0.05 to 5% by weight of the amount of soybean added, and the soybean inoculated with the strain is The soybean inoculated with the strain may be fermented at a temperature of 30 to 45 ° C and the soybean inoculated with the strain may be fermented for 1 to 5 days .

As used herein, the term "soybean fermented product" means a soybean fermented product produced by utilizing the fermentation action of microorganisms using soybean as a raw material. Examples of soybean fermented products include, but are not limited to, miso, kochujang, meju, .

In the present invention, diabetes mellitus is a disease in which insulin secretion or action of insulin on cells is decreased and absorption of glucose into the body does not normally occur. It involves hyperglycemia. It causes not only carbohydrate metabolism but also lipid and protein metabolism, Atherosclerosis, retinal hemorrhage, hypertension, and the like. Diabetes therapy and exercise therapy as well as pharmacotherapy are used in diabetes therapy. Drugs commonly used in clinical practice are insulin preparations, sulphonylurea preparations, biguanide preparations, troglitazone preparations, and alpha glucosidase inhibitors. Among them, the alpha glucosidase inhibitor is used for the prevention and treatment of diabetes because it decreases the decomposition rate of the small saccharides and disaccharides in the food to reduce the postprandial increase in blood glucose and the insulin reaction in the blood, This is an exemplary alpha glucosidase inhibitor.

In the present invention, 'Chungkukjang' refers to a fermented food in which soybean is fermented at a high temperature (40 ° C) after being called and steamed, and a bacterium belonging to the genus Bacillus is propagated to produce a sticky viscous substance. The Chungkukjang contains Bacillus spp. As a major microorganism.

As described above, the present invention provides a fermented soybean having antidiabetic activity due to a strain having high alpha glucosidase inhibitory activity.

Hereinafter, the present invention will be described in more detail with reference to Examples and Test Examples. It is to be understood, however, that the scope of the present invention is not limited to these embodiments.

Example

Example  1. Isolation and Identification of Strain

The strains were isolated from conventionally prepared kochujang using a tryptic case soybean agar (Difco, USA) and identified by 16S rDNA sequencing. As a result of identification, Bacillus methylotrophicus TPP 0025 ( Bacillus methylotrophicus TPP 0025, KCCM11279P) is trophy carcass (Bacillus in conventional Bacillus methyl methylotrophicus ), and the strain used in the present invention was deposited with KCCM on June 14, 2012.

Example  2. Manufacture of Chungkukjang

The soybean was washed for the preparation of Chongkukjang, and the soybean was soaked overnight, then the water was removed and the soybean was heated at 114 ° C for 50 minutes. After the addition, the strain was inoculated to 200 g of chilled soybeans so that the strain became 1% of the soybean yield and fermented in a thermostat at a temperature of 37 ° C and a humidity of 70% for 3 days to prepare chungkukjang. It was freeze-dried, pulverized, stored at -20 ° C, and the activity of alpha glucosidase inhibition was measured.

Example  3. Alpha-glucosidase  Measurement of inhibitory activity

10 g of the chungkukjang prepared according to Example 2 was added with 10 times of distilled water, followed by mixing at 200 rpm for 60 minutes. After heating for 10 minutes and centrifuging at 10,000 rpm for 10 minutes, the activity of alpha glucosidase inhibition of the supernatant was measured. To the 96 well plate was added 147 μL of the supernatant, 30 μL of 0.1 M phosphate buffer (pH 7.2), 30 μL of PNPG (1.42 mM) and 5 μL of alpha glucosidase (1.8 U / mL) . After 10 minutes, the reaction was stopped by adding 100 uL of 0.5 M Na ₂ CO ₃ and absorbance was measured at 405 nm. The inhibition rate was calculated by the following equation.

Inhibition rate (%) = [1- (A ₄₀₅ of sample-A ₄₀₅ of blank) / A ₄₀₅ of control] × 100

Strain Alpha Glucosidase Inhibitory Activity of Chungkukjang (%) Alpha glucosidase inhibitory activity (%) of the strain culture solution Production Example 1 Bacillus methylotrophicus TPP  . 77.5 40.3 Comparative Example 1 Bacillus subtilis subsp . subtilis 210 2.6 4.3 Comparative Example 2 Bacillus lichenifomis 248 5.0 6 Comparative Example 3 Bacillus amyloliquefaciens subsp. plantrum 21 11.4 2.7 Comparative Example 4 Bacillus methylotrophicus 386 30 0 Comparative Example 5 Bacillus subtilis subsp . inaquosorum 387 1.0 0 Control 1 Soybean soybeans 0.23 - Control group 2 Marketed Cheonggukjang 1 0.19 - Control group 3 Available Cheonggukjang 2 0.06 - Control group 4 Marketed Cheonggukjang 3 0.02 - Control group 5 Available Cheonggukjang 4 2.77 -

On the other hand, the commercial soybean, Chungkukjang 1, 2, 3, 4 and the soybeans not inoculated with the strain showed almost no inhibitory activity against alpha glucosidase, whereas Bacillus methylotrophicus TPP 0025 methylotrophicus TPP 0025, KCCM11279P) showed a high inhibition rate of alpha glucosidase of 77.5%. Bacillus subtilis 210, another strain isolated from traditional fermented foods such as soy sauce, meju, and salted fish, subsp . subtilis 210), Bacillus Kenny Li Po Miss 248 (Bacillus lichenifomis 248), Bacillus amyl Lori query Pacifico Enschede sub-Made Systems Planta Rum 21 (Bacillus amyloliquefaciens subsp . plantrum 21), Bacillus methylotrophicus 386) and Bacillus subtilis subspecies inactivosum 387 ( Bacillus < RTI ID = 0.0 > subtilis subsp . inaquosorum 387), the inhibitory activity of alpha glucosidase was found to be low. As a result, Bacillus methylotrophicus TPP 0025 methylotrophicus TPP 0025, KCCM11279P), the inhibitory activity of alpha glucosidase was remarkably increased.

Further, as a result of the measurement done methyl Bacillus alpha-glucosidase inhibitory activity of a strain culture solution itself trophy coarse TPP 0025 (Bacillus methylotrophicus TPP 0025, KCCM11279P) showed 40.3% inhibition activity of alpha glucosidase in the culture medium itself. Thus, it was confirmed that the strain itself has an activity of inhibiting alpha glucosidase and that its activity is further enhanced by preparing it with Chungkookjang.

Alpha glucosidase in the small intestinal mucosa is able to inhibit postprandial hyperglycemia because the saccharides degraded by alpha amylase are finally converted into monosaccharides. Therefore, a substance having an alpha glucosidase inhibitory activity can be used for prevention and treatment of diabetes. Thus, Bacillus methylotrophicus TPP 0025 ( Bacillus < RTI ID = 0.0 > methylotrophicus TPP 0025, KCCM11279P) and chungkukjang prepared with this strain have high alpha-glucosidase inhibitory activity and can effectively inhibit the formation of monosaccharides in the digestion process of carbohydrate, so that it can be used for prevention and treatment of diabetes through postprandial glucose control .

While the present invention has been described with reference to exemplary embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof without departing from the scope of the invention. You will know. In addition, many modifications may be made to adapt a particular situation and material to the teachings of the invention without departing from the essential scope thereof. Accordingly, it is intended that the invention not be limited to the particular embodiment disclosed as the best mode contemplated for carrying out this invention, but that the invention be construed as including all embodiments falling within the scope of the appended claims.

Korea Microorganism Conservation Center (overseas) KCCM11279P 20120614

Claims (14)

Bacillus < / RTI > methylotrophicyst TPP 0025 ( Bacillus < RTI ID = 0.0 > methylotrophicus TPP 0025, KCCM11279P). Inoculating soybeans supplemented with Bacillus methylotrophicus TPP 0025 (KCCM11279P) strain; And
And fermenting the soybeans inoculated with the strain to produce soybean fermented product having enhanced antidiabetic effect. delete 3. The method of claim 2,
Wherein the strain is inoculated with 0.05 to 5% by weight of the amount of soybeans added. 3. The method of claim 2,
Wherein the soybean inoculated with the strain is fermented under a condition of a humidity of 60 to 80%. 3. The method of claim 2,
Wherein the soybean inoculated with the strain is fermented at a temperature of 30 to 45 캜. 3. The method of claim 2,
Wherein the soybean inoculated with the strain is fermented for 1 to 5 days. A soybean fermented product having enhanced antidiabetic effect including a strain of Bacillus methylotrophicus TPP 0025 (KCCM11279P) and a soybean grown therein. delete 9. The method of claim 8,
Wherein the strain is 0.05 to 5% by weight of the amount of soybeans added. 9. The method of claim 8,
Wherein the soybean inoculated with the strain is fermented under a condition of a humidity of 60 to 80%. 9. The method of claim 8,
Wherein the soybean inoculated with the strain is fermented at a temperature of 30 to 45 占 폚. 9. The method of claim 8,
Wherein the soybean inoculated with the strain is fermented for 1 to 5 days. A food comprising soybean fermented product produced according to the method of any one of claims 2 and 4 to 7, wherein the soybean fermented product has been improved.