JP2001213773A - Medicine for improving hyperetension and diabetes and production of gamma aminobutyric acid - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method of producing γ-aminobutyric acid(GABA) with several times productivity compared with the method used previously buy cultivating embryos and Ascomycetes, and also to provide a medicine for improving hypertension and diabetes having γ-aminobutyric acid as an effective main component. SOLUTION: This medicine contains γ-aminobutyric acid produced by Basidiomycetes belonging to the genus Agaricus of the hampignon family as the main component.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a physiologically active substance produced by agaricaceae basidiomycetes, which is particularly used as a health food, and more particularly to an agent for improving hypertension and diabetes and a method for producing γ-aminobutyric acid. .

[0002]

BACKGROUND ART Agaricus genus Basidiomycetes is currently
It is used in health foods and the like as an immunostimulant, an anticancer agent, and the like. Its active ingredient is presumed to be derived from carbohydrates in bacterial cells in immunostimulants and anticancer drugs.

[0003] However, there have been no reports on the effects of improving hypertension and the effect of improving diabetes derived from amino acids produced by the genus Agaricus.

On the other hand, it is known that some ascomycetes produce γ-aminobutyric acid, which is known as a neurotransmitter, and are commercially used as health foods.
Basidiomycetes have not been reported to produce significant amounts of γ-aminobutyric acid.

It is known that cereal germ and tea leaf enzymes convert glutamic acid to γ-aminobutyric acid, but some basidiomycetes perform similar conversion to produce a significant amount of γ-aminobutyric acid.
It is not known to produce aminobutyric acid.

[0006]

However, during the various studies on the bioactive substances of basidiomycetes, the present inventors have found that the basidiomycete Agaricus sp. Has a higher concentration of γ-amino acid than these ascomycetes. The inventors have found that it produces butyric acid and have invented a method for significantly increasing the production of γ-aminobutyric acid.

[0007] γ-aminobutyric acid is known as a neurotransmitter, has an effect of improving hypertension, and it is widely known that deficiency in γ-aminobutyric acid production is associated with the development of certain types of diabetes. Although it has been shown that the production of γ-aminobutyric acid without chemical synthesis is possible by germ and ascomycete cultivation, it is commercially only performed on a small scale because of its low productivity. Absent.

[0008] The present invention provides a method for producing γ-aminobutyric acid (GABA) with a productivity several times higher than that of a conventional method using embryo and ascomycete culture, and also provides γ-aminobutyric acid (GABA).
-To provide an agent for improving hypertension and diabetes containing aminobutyric acid as an active ingredient.

[0009]

The basidiomycetes used in the present invention are Agaricus blazei Murill and Agaricus bisporus.

[0010] The medium used for liquid culture is a carbon source.
Barley, wheat, rice, corn and other rhizomes, potatoes, sweet potatoes, cassava and other rhizomes, or starches derived from them, or their decomposed products, glucose, sucrose, fructose and other monosaccharides, small sugars, nitrogen As a source, inorganic nitrogen such as ammonium sulfate and organic nitrogen such as shochu residue, corn steep liquor, yeast extract, protein decomposed product, amino acid, cyanide and the like may be used alone or in combination. Minerals and vitamins may be added according to the proportion of these natural products.

As a device used for the culture, a usual aeration-stirred culture tank or an air-lift type culture tank is used.

The culture medium is sterilized in a culture tank or sterilized or disinfected outside the tank, and is subjected to batch culture. The culture is added to the medium sterilized or disinfected outside the tank, and the culture is performed while removing the product. To perform continuous culture.

Further, in order to increase the production of γ-aminobutyric acid, 1 to 50 g / l of sodium glutamate is added to the medium. This may be from the beginning of the culture, or during or after the end of the culture, and before the solid content separation of the culture.

[0014]

Embodiments of the present invention will be described below.

Agaricus blazei M. was used as a test plot, Aspergillus oryzae was cultured as a control plot 1, and Monascus sp. As a control plot 2.

The components of the medium used for the culture are as shown in Table 1.

The shochu residue used in the medium was obtained by removing most of the solid content from the distillation residue of barley shochu using a centrifugal decanter. The components are shown in Table 2. Mineral is K ₂
HPO ₄ : 80%, MgSO ₄ .7H ₂ O: 20%.

The medium is mixed with tap water, sterilized at 120 ° C. for 20 minutes in a 10-liter fermenter, allowed to cool, and inoculated with an inoculum.
The cells were cultured at 25 ° C. for 11 days. The initial pH was 5.0. The amount of γ-aminobutyric acid produced is as shown in Table 1. Table 1 also shows the residual amount of glutamic acid, which is a precursor of γ-aminobutyric acid, and the ratio of γ-aminobutyric acid / glutamic acid.

[0019]

[Table 1] Test plot Control plot 1 Control plot 2 Components Shochu distillation residue (ml / l) 300 300 300 Glucose (g / l) 50 50 50 Mineral (g / l) 3 3 3 Thiamine hydrochloride (mg / l) ) 10 10 10 Amount of γ-aminobutyric acid produced (mg / l) 360.7 243.9 146.6 Residual glutamic acid (mg / l) 60.1 557.5 646.0 Ratio 6.00 0.44 0.23 As shown in Table 1, Agaricus blazei M. produced γ-aminobutyric acid. Glutamic acid utilization is high. The liquid classification of the shochu distillation residue used above is shown in Table 2 below.

[0020]

[Table 2] Solid content 8.8% Crude protein (Protein coefficient: 6.14) 4.18% Total sugar 2.45% Straight sugar 0.54% Crude fat 0.32% pH 4.0 Total polyphenol 0.40% Dietary fiber 2.46% Amount required for the growth of Agaricus blazei M. Glutamic acid was supplied in excess to culture.

The components of the medium used for the culture are as shown in Table 3.

The mineral composition in Table 3 is K ₂ HPO ₄ : 3
_{7.7%, KH 2 PO 4:} 37.7%, MgSO 4 · 7H 2
0: 18.9%, CaCl ₂ .2H ₂ O: 5.0%, Fe
_{SO 4 · 7H 2 0: 0.4} %, ZnSO 4 · 7H20:
_{0.1%, MnCl 2 · 4H 2} O: was 0.2%.

The medium was mixed with tap water, sterilized at 120 ° C. for 20 minutes in a 10-liter fermenter, allowed to cool, and inoculated with a seed bacterium.
The cells were cultured at 25 ° C. for 11 days. The initial pH was 5.0. The production amount of γ-aminobutyric acid is as shown in Table 3.

[0024]

Table 3 Test plot 1 Test plot 2 Component Yeast extract (g / l) 55 (Oriental yeast) Glucose (g / l) 50 50 Mineral (g / l) 33 Thiamine hydrochloride (mg / l) 10 10 Sodium glutamate (g / l) 0 10 Production amount of γ-aminobutyric acid in culture solution (mg / l) 149 2081 Production amount of γ-aminobutyric acid in bacterial cells (mg / 100 g. Dried product) 196 2164 As shown in Table The addition of sodium glutamate significantly increased the γ-aminobutyric acid content inside and outside the cells. Agaricu obtained from the test plot 2 of the culture indicated in 2
Table 4 shows the analytical values of the γ-aminobutyric acid content of the dried product obtained by drying and diluting the s blazei M. culture and the commercially available γ-aminobutyric acid-containing natural product.

For the culture, dextrin was dispersed so that the content of γ-aminobutyric acid in the culture solution was 1%, followed by drying under reduced pressure, and the cells were directly dried under reduced pressure.

[0026]

[Table 4] Content of γ-aminobutyric acid (mg / 100g) ORYZA GABA GERM P (Oriza Yuka Co., Ltd.) 279 (from rice germ) ORYZA GABA EXTRACT C (ORYZA YUKA CORPORATION) 407 (from rice germ) Agaricus blazei M. Dried culture solution (after addition of dextrin) 1,000 Agaricus blazei dried cells 2,164

[0027]

EXAMPLES Examples of the present invention will be described below in more detail.

Example 1 Agaricus blazei M. was cultured by donating glutamic acid in excess of the amount required for growth.

The components of the medium used in the culture are as shown in the table.

The shochu residue used in the culture medium was obtained by removing most of the solid content from the distillation residue of barley shochu by a centrifugal decanter. The components are shown in Table 5. Mineral is K ₂
HPO ₄ : 80%, MgSO ₄ .7H ₂ O: 20%.

The medium was mixed with tap water, sterilized in a 10-liter fermenter at 120 ° C. for 20 minutes, allowed to cool, and inoculated with a seed bacterium.
The cells were cultured at 25 ° C. for 11 days. The initial pH was 5.0.

[0032]

Table 5 Ingredients Shochu distillation residue (ml / l) 300 Glucose (g / l) 50 Mineral (g / l) 3 Thiamine hydrochloride (mg / l) 10 Sodium glutamate (g / l) 10

[0033]

[Table 6] Solid content 8.8% Crude protein (Protein coefficient: 6.14) 4.18% Total sugar 2.45% Straight sugar 0.54% Crude fat 0.32% pH 4.0 Total polyphenol 0.40% Dietary fiber 2.46% After cultivation, No.2 filter paper and suction Perform solid-liquid separation with a filter,
The filtrate was concentrated under reduced pressure using a rotary evaporator,
Freeze vacuum drying was performed as it was. The filtration residue was directly dried under reduced pressure. The obtained dried solid content was 40 g / l from the culture solution, and the yield of cells was 10 g / l.

The content of γ-aminobutyric acid in the dry solid content was 5 g / 100 g from the filtrate, and 2 g / 100 g from the filtration residue.
Met.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) C12P 13/00 C12P 13/00 // (C12N 1/14 (C12N 1/14 E C12R 1: 645) C12R 1: 645) (C12P 13/00 (C12P 13/00 C12R 1: 645) C12R 1: 645) (72) Inventor Tapan Kumar Mazmuda 681, Rokutaya, Hayashida-cho, Himeji City, Hyogo Prefecture In-house F-term (reference) 4B018 MD18 ME03 ME04 4B064 AE01 CA07 CD13 CD22 CD24 DA01 DA10 4B065 AA71X AC14 BB12 BB26 BB29 CA16 CA41 4C206 AA01 AA02 AA04 FA45 MA01 MA04 NA14 ZA42 ZC35

Claims (6)

[Claims] 1. An agent for improving hypertension and diabetes, comprising as a main component γ-aminobutyric acid produced by Agaricus basidiomycete, Agaricaceae. 2. The agaric basidiomycete is Agaricus blaz.
The hypertensive and diabetes improving agent according to claim 1, which is ei Murill. 3. A method for producing γ-aminobutyric acid, wherein γ-aminobutyric acid is produced by subjecting a basidiomycete belonging to the genus Agaricus belonging to the genus Agaricus to liquid culture in a liquid medium containing glutamic acid or glutamate. . 4. The liquid medium includes, as a carbon source, cereals such as barley, wheat, rice, and corn; rhizomes such as potato, sweet potato, and cassava; starches made from these as raw materials; decomposed products thereof; glucose; and sucrose. , Monosaccharides such as fructose, and small sugars, and inorganic nitrogen such as ammonium sulfate as a nitrogen source;
The method for producing γ-aminobutyric acid according to claim 3, wherein organic nitrogen such as shochu residue, corn steep liquor, yeast extract, protein decomposed product, amino acid, and cyanide is used alone or in combination. 5. The method for producing γ-aminobutyric acid according to claim 3, wherein the liquid medium contains glutamic acid or glutamate in a total amount of 6 to 300 mmol / l. 6. The γ-protein according to claim 3, wherein 1 to 50 g / l of sodium glutamate is added to the liquid medium.
A method for producing aminobutyric acid.