KR102253265B1 - Method for producing powder of fermented Protaetia brevitarsis larva with increased poly gamma glutamic acid and powder of fermented Protaetia brevitarsis larva with increased poly gamma glutamic acid produced by the same method - Google Patents

Abstract

The present invention relates to a method for producing a powder of fermented white spotted radish larva with an enhanced content of poly-gamma-glutamic acid, and a powder of fermented white spotted radish larva with an enhanced content of poly-gamma-glutamic acid prepared by the method. Compared to the white spotted radish larva powder, the powder of plain larvae fermentation not only contains a large amount of polygamma-glutamic acid, but also has excellent reducing effects of ALT, AST, and ALP, and does not cause side effects because it uses substances obtained from natural products. It is expected to be very useful in functional food and pharmaceutical industries for functional improvement.

Description

Method for producing powder of fermented white-spotted radish larvae with enhanced poly-gamma-glutamic acid content, and powder of fermented white-spotted radish larvae with increased poly-gamma-glutamic acid content prepared by the above method. increased poly gamma glutamic acid and powder of fermented Protaetia brevitarsis larva with increased poly gamma glutamic acid produced by the same method}

The present invention relates to a method for producing a powder of fermented white-spotted radish larvae with enhanced poly-gamma-glutamic acid content and a powder of fermented white-spotted radish larvae with increased poly-gamma-glutamic acid content prepared by the method.

Protaetia brevitarsis is known as'manufacturing'or'slug' in traditional oriental medicine books such as private and Donguibogam (Coleoptera), Scarabaeidae, and flower. It is a phytophagous insect with a size of 17 to 24 cm belonging to the Cetoniinae family. It is known to live in China, Japan, and eastern Siberia, including Korea. The white-spotted flower radish winters in the state of 3 instar mature larvae, and adults are active during the day and feed on mature fruits such as peaches and pears, or juices such as corn and oak trees, and are always clustered. In addition, their larvae live in corrosive soils rich in organic matter such as compost and haystacks.

White spotted radish has long been used as a herbal medicine for the treatment of liver disease and the like. In addition, recently, it has attracted great attention as an insect resource for the search and development of useful biologically active substances, and the production of antibiotics and the recovery effect of interlipid metabolism damaged by excessive alcohol intake in experiments using mice. It is known, and its usefulness has been confirmed, including studies on thrombolytic enzymes and liver protection against hepatotoxicity induced by the administration of carbon tetrachloride in rats.

Meanwhile, poly gamma glutamic acid (PGA) is a polymer in which D,L-glutamic acid is combined with gamma-glutamyl, and is a mucous substance. Polygamma-glutamic acid is produced from Bacillus strains isolated from Korean traditional fermented soybean food using rice straw, Cheonggukjang, Japanese traditional fermented soybean food, Natto, and Nepalese traditional fermented soybean food, Kinema. Poly-gamma-glutamic acid produced from the strains of the genus Bacillus is an edible, water-soluble, anionic, biodegradable polymer material that can be used as a moisture absorbent, moisturizer and raw material for cosmetics.

On the other hand, Korean Patent No. 1667403 discloses a'food stabilizer composition containing poly-gamma-glutamic acid', and Korean Patent No. 0475406 discloses'a composition for enhancing immunity containing poly-gamma-glutamic acid', but the present invention There is no description of the powder manufacturing method of the fermented white spotted radish larva with the enhanced content of poly-gamma glutamic acid and the powder of the fermented white spotted radish larva with the enhanced poly-gamma-glutamic acid content prepared by the above method.

The present invention was derived from the above requirements, and the present inventors prepared a soybean fermented product by mixing a Bacillus subtilis strain with soybeans, and then mixed the soybean fermented product with white spotted radish larva powder to maintain moisture. To prepare a fermented product of white spotted radish larva by adding sterile water for solid phase fermentation, the fermented product was dried and pulverized to prepare a powder of fermented white spotted radish larvae. As a result of quantitative analysis of the prepared white spotted radish larvae fermented product using an automatic amino acid analyzer, the content of poly-gamma-glutamic acid in the powder of white-spotted radish larvae fermented product was about as compared to the non-fermented white-spotted radish larvae powder. It was confirmed that it increased by 38%. In addition, when the powder of fermented white spotted radish larvae was fed to the animal model of non-alcoholic fatty liver, the content of ALT, AST, and ALP in the serum decreased compared to the group fed with the non-alcoholic fatty liver-causing group and the white spotted radish larva powder. By confirming that the powder of fermented white spotted radish larvae is effective in improving liver function, the present invention was completed.

In order to solve the above problems, the present invention comprises the steps of: (1) inoculating and culturing a strain of Bacillus subtilis in sterile water containing soybeans to prepare a soybean fermentation product; (2) The soybean fermented product of step (1) was mixed with white spotted flower radish ( Protaetia). brevitarsis ) preparing a mixture by mixing with larva powder, and then adding water to the mixture to ferment in a solid phase to prepare a fermented product of white spotted radish larva; And (3) drying and pulverizing the fermented product of white spotted radish larvae of step (2) after high pressure sterilization; fermenting white spotted radish larvae with enhanced content of poly gamma glutamic acid. It provides a method for producing a powder of water.

In addition, the present invention provides a powder of fermented white spotted radish larva with an enhanced content of poly-gamma-glutamic acid prepared by the above manufacturing method.

In addition, the present invention provides a health functional food and pharmaceutical composition for improving liver function comprising the powder of the fermented white spotted flower radish larvae as an active ingredient.

The powder of the fermented white spotted radish larvae of the present invention not only contains a large amount of polygamma-glutamic acid, but also reduces the concentration of ALT (alanine aminotransferase), AST (aspartate aminotransferase) or ALP (lactate dehydrogenase), thereby improving liver function. Since it is excellent, it will be useful as a health functional food and pharmaceutical composition for improving liver function.

1 is a schematic diagram of a powder manufacturing process of fermented white spotted radish larvae.
Figure 2 is a graph analyzing the content of poly-gamma-glutamic acid in the powder of non-fermented white spotted radish larvae powder and white spotted radish larvae fermented product.

In order to achieve the object of the present invention, the present invention

(1) preparing a soybean fermented product by inoculating and culturing a Bacillus s ubtilis strain in sterile water containing soybeans;

(2) After mixing the soybean fermented product of step (1) with the white spotted radish (Protaetia brevitarsis) larva powder to prepare a mixture, water was added to the mixture to solidify fermentation to prepare the white spotted radish larvae fermented product. Manufacturing steps; And

(3) the step of autoclaving the fermented product of white spotted radish larvae of step (2), drying and pulverizing; It provides a method for producing a powder.

In the method according to an embodiment of the present invention, the method of producing a powder of fermented white spotted radish larva with an enhanced poly-gamma-glutamic acid content is more specifically,

(1) preparing a soybean fermented product by inoculating and culturing a Bacillus subtilis strain in sterile water containing 3-7% (w/v) soybeans;

(2) The soybean fermented product of step (1) was added to the white spotted radish larva powder and 5 to 15% (v/w) based on the weight of the powder was added and mixed to prepare a mixture, and then water based on the weight of the mixture was added to the mixture. Adding 50-60% and performing solid-state fermentation at 28-32° C. for 70-74 hours to prepare a fermented product of white spotted radish larvae; And

(3) drying and pulverizing the white spotted radish fermented product of step (2) at 115 to 125° C. for 30 to 50 minutes, followed by drying and grinding; may include, and

More specifically,

(1) preparing a soybean fermented product by inoculating and culturing a Bacillus subtilis strain in sterile water containing 5% (w/v) soybeans;

(2) The soybean fermented product of step (1) was added to the white spotted radish larva powder and 10% (v/w) based on the weight of the powder was added and mixed to prepare a mixture, and then water based on the weight of the mixture was added to the mixture. Adding 60% and performing solid-state fermentation at 30° C. for 72 hours to prepare a fermented product of white spotted radish larva; And

(3) drying and pulverizing the white spotted radish fermented product of step (2) at 121° C. for 30 to 50 minutes, followed by drying, but is not limited thereto.

The present invention also provides a powder of fermented white spotted radish larva with an enhanced content of poly-gamma-glutamic acid prepared by the above manufacturing method.

The powder of the white spotted radish larvae fermented product increased the poly-gamma-glutamic acid content in the white-spotted radish larva powder by about 35-40% compared to the non-fermented white-spotted radish powder.

The present invention also provides a health functional food composition for improving liver function comprising the powder of fermented white spotted radish larva with an increased PGA content as an active ingredient.

The health functional food composition of the present invention is a fermented product of white spotted radish larvae having excellent effect of reducing the content of ALT (alanine aminotransferase), AST (aspartate aminotransferase) and ALP (alkaline phosphatase) in serum through a fermentation process by Bacillus subtilis. Since the powder is included as an active ingredient, there is an effect of improving liver function.

The health functional food composition for improving liver function may be prepared in any one formulation selected from powders, granules, pills, tablets, capsules, candy, syrup, effervescent tablets, and beverages, but is not limited thereto. The health functional food composition is not particularly limited as long as it can be ingested to prevent or relieve a hangover.

When the powder of the fermented white spotted radish larva of the present invention is used as a food additive, the powder of the fermented white spotted radish larvae may be added as it is or may be used with other foods or food ingredients, and appropriately according to a conventional method. Can be used. The active ingredient may be appropriately used depending on the purpose of use (prevention or improvement). In general, when preparing food or beverage, the fermented product of the aged sprout ginseng extract of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less based on the raw material. However, in the case of long-term intake for the purpose of health control, it can be used in an amount within a range without any problems in terms of safety.

There is no particular limitation on the type of the food. Examples of foods to which the health functional food composition can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea There are drinks, alcoholic beverages, and vitamin complexes, and all health foods in the usual sense are included. The beverage includes carbonated beverages, functional ionic beverages, juices (eg, apples, pears, grapes, aloe, tangerines, peaches, carrots, tomato juice, etc.), sikhye, and the like.

Functional foods of the present invention include ingredients that are commonly added during food production, and include, for example, proteins, carbohydrates, fats, nutrients, and seasonings. For example, when prepared as a drink, natural carbohydrates or flavoring agents may be included as an additional ingredient in addition to the active ingredient. The natural carbohydrates are monosaccharides (e.g., glucose, fructose, etc.), disaccharides (e.g., maltose, sucrose, etc.), oligosaccharides, polysaccharides (e.g., dextrins, cyclodextrins, etc.) or sugar alcohols (e.g. , Xylitol, sorbitol, erythritol, etc.). As the flavoring agent, natural flavoring agents (eg, taumatin, stevia extract, etc.) and synthetic flavoring agents (eg, saccharin, aspartame, etc.) may be used. In addition to the health functional food composition, various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonic acid It may further contain a carbonation agent used in beverages.

Hereinafter, the present invention will be described in detail by examples. However, the following examples are merely illustrative of the present invention, and the contents of the present invention are not limited to the following examples.

Materials and methods

One. White spotted flower U insect Fermented Powder manufacturing

Bacillus subtilis (Bacillus subtilis KACC 91157) was inoculated in sterile water containing 5% (w/v) of soybeans and cultured at 30° C. and 100 rpm for 24 hours to prepare a soybean fermented product. White spotted radish larva powder is sterilized with a high pressure sterilizer at 121°C for 15 to 30 minutes and naturally cooled, and then the soybean fermented product was added to 1 kg of the naturally cooled white spotted radish larva powder by 10% (v/ w) Add and mix, add 50-60% of sterilized water based on the weight of the mixture to the mixture, ferment in solid phase at 30℃ for 72 hours, and then autoclave at 121℃ for 30-50 minutes to terminate the fermentation. Plain fermented larvae were prepared. The prepared white spotted radish larva fermented product was dried in a dry oven and pulverized to prepare a powder of white spotted radish larvae fermented product (FIG. 1). It was confirmed with the naked eye that polygamma-glutamic acid, a sticky viscous substance, was produced on the powder surface of the fermented white spotted radish larvae.

2. Experimental animals

SD (Sprague-Dawley) rats (6 weeks old, male) were classified using a randomized block design (randomized block design) so that the average weight was the same, and 6 to 8 rats were classified for each experimental group. Temperature 22±2℃, humidity 50±5% And it was reared in an animal breeding room controlled at 12 hours of light and dark cycles. It was adapted to the environment by feeding a commercial solid feed for a week.

3. Serum analysis of experimental animals

The rats were fed a formula (Table 1) for 10 days to induce a normal animal model and a non-alcoholic fatty liver animal model, and the rats fed a fatty liver-induced diet were fermented with white spotted radish larva powder or white spotted radish larva. The water powder was mixed and fed (Table 2). On the last day of the experiment, rats were fasted for 12 hours and then dissected by anesthetizing with diethyl ether. Blood was collected from the abdominal aorta after laparotomy, allowed to stand at room temperature for about 30 minutes, and then centrifuged at 3,000 rpm for 20 minutes to obtain serum. The content of ALT (alanine aminotransferase), AST (aspartate aminotransferase), and ALP (alkaline phosphatase) in serum was analyzed by Dongnam Medical Chemical Research Institute, a medical examination agency.

Formulated diet scheme ingredient Normal diet (%) Fatty liver induction diet (%) Casein 20.0 20.0 Corn starch 15.0 14.0 Corn oil 10.0 10.0 Cellulose 5.0 5.0 Mineral mixture 4.0 4.0 Vitamin mixture 1.0 1.0 L-methionine 0.3 0.3 Sucrose 44.5 44.5 Choline 0.2 0.2 Orotic acid - 1.0 Sum 100 100

Experimental sample dose Experimental group ingredient N Normal diet OA Normal diet + fatty liver induction diet OA+NPb Fatty Liver Induction Diet + NPb 100 mg/kg of b.w./day OA+FPb Fatty liver induced diet + FPb 100 mg/kg of b.w./day

N; Normal.

OA; Non-alcoholic fatty liver induced group.

OA+NPb; Experimental group in which non-alcoholic fatty liver was induced and white spotted radish larva powder was fed.

OA+FPb; Experimental group in which non-alcoholic fatty liver was induced and powder of fermented white spotted radish larva was fed.

Example One. White spotted flower larva Fermented Of powder Polygammaglutamic acid Quantitative analysis of content

After setting poly-gamma-glutamic acid as an indicator of the powder of the fermented white-spotted larva through the mucus formed on the powder surface of the fermented white-spotted larva, an automatic amino acid analyzer was used to quantitatively analyze it. The analysis method used the poly-gamma-glutamic acid test method notified by the Ministry of Food and Drug Safety.

As a result, the content of poly-gamma-glutamic acid in the non-fermented white spotted radish larva powder was 47.17 mg/g, and the poly-gamma-glutamic acid content of the powder of white spotted radish larvae fermented was 50.31 mg/g on the first day of fermentation, and fermentation 2 It was confirmed that it was 64.74mg/g on the first day and 65.50mg/g on the third day of fermentation. In particular, the content of poly-gamma-glutamic acid was increased from the second day of fermentation of the powder of the fermented white spotted radish larvae, and it was found that on the third day of fermentation, the content of polygamma-glutamic acid increased to about 38% compared to before fermentation (FIG. 2).

From the above results, it was considered that Bacillus subtilis, which has the property of producing protease, proliferates on the second day of fermentation using the white-spotted radish larva containing 50% or more of protein as a substrate and enters a stable phase on the third day.

Example 2. White spotted flower larva Fermented The effect of improving liver function of powder

In rats with non-alcoholic fatty liver, ALT, AST and ALP contents in mouse serum were analyzed to confirm the effect of improving liver function of the powder of fermented white spotted radish larvae.

As a result, ALT, AST, and ALP contents were all increased in the non-alcoholic fatty liver inducer group (OA) compared to the normal group (N), and the white spotted radish larva powder or fermented white-spotted radish larva in the non-alcoholic fatty liver-induced group The ALT, AST and ALP contents were all decreased in the group fed with the powder compared to the non-alcoholic fatty liver-induced group. In particular, it was confirmed that the group fed the powder of the white spotted radish larvae fermented product decreased ALT, AST, and ALP contents more than the group fed the white spotted radish larvae powder, which was similar to that of the normal group. Through this, it was found that fermenting the white spotted radish larva powder can further increase the effect of improving liver function (Table 3).

Measurement of ALT, AST and ALP content in serum ALT(U/L) AST(U/L) ALP(g/dL) N 33.4 120.2 157.0 OA 41.0 134.7 193.0 OA+NPb 36.9 110.2 172.0 OA+FPb 34.9 99.1 156.7

Claims (5)

delete delete delete As a health functional food composition for improving non-alcoholic fatty liver, comprising the powder of fermented white spotted radish larvae with an enhanced content of poly gamma glutamic acid as an active ingredient,
The powder of the fermented white spotted radish larva with an enhanced content of poly gamma glutamic acid,
(1) preparing a soybean fermented product by inoculating and culturing a Bacillus subtilis strain in water containing 3-7% (w/v) soybeans;
(2) The soybean fermented product of step (1) was added to the white spotted flower radish (Protaetia brevitarsis ) larva powder and 5-15% (v/w) based on the weight of the powder was added and mixed to prepare a mixture, and then the mixture was mixed with the mixture. Preparing a fermented product of white spotted radish larvae by adding 50-60% of water by weight and fermenting in a solid state at 28-32°C for 70-74 hours; And
(3) the step of autoclaving the fermented white spotted flower radish larvae of step (2) at 115 to 125° C. for 30 to 50 minutes, followed by drying and pulverizing; and
A health functional food composition for improving non-alcoholic fatty liver, characterized in that reducing the content of ALT (alanine aminotransferase), AST (aspartate aminotransferase) and ALP (alkaline phosphatase). delete

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