JPH0634741B2 - How to make nosyheptide - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to a method for producing nosiheptide using microorganisms. Nociheptide is a sulfur-containing polypeptide antibiotic, and is used as a feed additive for the purpose of promoting animal growth, improving feed efficiency, and the like.

[Conventional technology]

For the production method of nosiheptide, for example, Streptomyces actuosus 400
37, NRRL-2954) culture method (Japanese Patent Publication No. 40-
No. 880) is known. Examples of microorganisms that produce nociheptide include Streptomyces antibioticus 8446-CC1 (Streptomyces antibioticus) and Streptomyces glaucoglyceus NRRL12514 (Strept
omyces glaucogriseus, NRRL12514), Actinoplanes SP NO.C-21740 (Actinoplanes SP NO.C-21740,
IFO-14018) is also known.

When accumulating nosiheptide in the medium by culturing these nosiheptide-producing microorganisms, in addition to ordinary carbon sources and nitrogen sources, Mg, Mn, Fe, Cu, Co, etc. should be performed in a medium containing a trace amount of metal. Have been reported (Japanese Patent Publication No. 40-8)
80, Experientia 36 , 414 (1980)).

[Problems to be solved by the invention]

However, it has been difficult to achieve sufficient accumulation of nosiheptide in the conventional medium as described above.

As a result of various studies on a method for increasing the productivity of nociheptide, we found that productivity was improved by culturing a nociheptide-producing bacterium in a medium containing a protein extracted and purified from soybean, and to complete the present invention. It arrived.

[Means for solving problems]

The gist of the present invention is to cultivate a microorganism belonging to the genus Streptomyces and / or Actinoplanes having the ability to produce nociheptide in a medium containing a protein extracted and purified from soybean, and produced / accumulated in the medium. And a method for producing a nociheptide characterized by collecting nosiheptide.

The present invention will be described in detail below.

First, the microorganism used in the present invention is a microorganism that belongs to actinomycetes and has the ability to produce nociheptide. Streptomyces actuosus 40037, N is a microorganism that produces nosiheptide.
RRL-2954), Streptomyces antibiotics 844
6-CC1 (Streptomyces antibioticus), Streptomyces glaucus NRRL12514 (Streptomyces glauc
ogriseus NRRL12514), Actinoplanes SP NO.C-21740
(Actinoplanes SP NO.C-21740, IFO-14018) and the like.

As a growth medium for the microorganism used in the present invention, any medium containing a nutrient source necessary for growth can be used. Carbon sources include glucose, fructose, maltose, sucrose, lactose, molasses, starch,
Examples thereof include sugars of dextrin sugar, sugar alcohols, carbohydrates such as glycerin and mannitol, organic acids such as lactic acid, citric acid or tartaric acid, and animal / vegetable oils such as lard oil and soybean oil. As the nitrogen source, soybean powder, cottonseed meal, dried yeast, soybean protein, peptone, corn stay liquor,
Organic and inorganic substances such as D. Taylors solve, fish meal, ammonium sulfate, ammonium salt, ammonium phosphate, sodium nitrate and urea can be used.

In the method of the present invention, it is necessary to further add a protein extracted and purified from soybean (hereinafter sometimes referred to as purified soybean protein) to a medium containing these. The refined soybean protein used is obtained by extracting oils and fats from soybeans, removing fibrin, carbohydrates, ash, etc., and effectively extracting only the protein.
Many products such as "AL" (made by Fuji Piulina Protein Co., Ltd.), "ESSAN PROTEIN F", "Adipron M-2" (made by Ajinomoto Co., Ltd.) are available on the market. It is processed into various forms such as granular and fibrous, but any form can be used.Preferably, when these purified soybean proteins are added to the medium at an appropriate ratio together with the above-mentioned other nitrogen sources, Nociheptide production is significantly improved.The amount added is determined by the protein content of the purified soybean protein, converted to protein, and 0.2 to 4% is added to the medium. (Purified soy protein /
It is desirable to add other nitrogen sources within the range of 0.1 to 3. Soybean flour is suitable as another nitrogen source.

If necessary, phosphates and salt may be added to the medium. If necessary, yeast extract, vitamins, nucleic acids, amino acids and the like may be added as micronutrients. As the sulfur source, sulfates, thiosulfates, cysteine, methionine, sulfocysteine, etc. are used.

It is desirable to carry out the culture under aerobic conditions, and it is generally advantageous to carry out aeration stirring culture and shaking culture. The culturing temperature is usually within the range of 20 to 40 ° C., but preferably 2
It is in the range of 5 to 33 ° C. The pH of the medium can be in the range of 4.5 to 9.0, but is preferably in the range of 6.0 to 8.0.

When an antifoaming agent is required, a commonly used defoaming agent can be added.

After completion of the culture, the desired product, nosiheptide, can be collected from the culture medium by using a solvent in accordance with a conventional method, for example, the method described in Japanese Patent Publication No. 40-880. In addition, for qualitative and quantitative determination of nosiheptide, automatic colorimetric analysis, bioassay by Staphylococcus aureus FDA 209P, thin layer chromatography, high performance liquid chromatography, infrared absorption spectrum, nuclear magnetic resonance, mass spectrometry, etc. Can be used.

〔Example〕

Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited to the following Examples unless the gist thereof is exceeded. In addition, "%" shows "weight%."

Defatted soybean powder 8.36%, Day Taylor Sorble 0.5
%, 2% soybean _{oil, 0.5% NaCl, MgSO 4 ·} 7H 2 O 0.05%, MnSO 4 ·
H ₂ O 0.03%, FeSO ₄ / 7H ₂ O 0.02%, CuSO ₄ / 5H ₂ O 0.002%, CoCl ₂
35 ml of a medium containing 0.002% of 6H ₂ O and a protein extracted and purified from soybean in the amount shown in Table 1 was added to a 200 ml flask and sterilized at 121 ° C. for 20 minutes, and Streptomyces actusosus ( Streptomyces actuosus ATCC
25421) was inoculated with 1 platinum loop and cultured at 27 ° C. and 210 RPM for 5 days. After culturing, the pH was adjusted to 8.5, diluted 2-fold with water, and then extracted by adding an equal amount of ethanol. After the extract was analyzed by bioassay, nosiheptide shown in Table 1 was produced.

[Comparative example]

Nociheptide was produced in the same manner as in Example 1 except that defatted soybean flour was added in the amount shown in Table 1 (in addition to the above 8.36%) instead of the protein extracted and purified from soybean. The results are shown in Table 1.

[Effect of the Invention] According to the method of the present invention, by using the protein extracted and purified from soybean, the productivity of nociheptide can be improved as compared with the case of using other nitrogen sources having the same protein amount.

Claims (1)

[Claims] 1. A nociheptide produced and accumulated in a culture medium containing a protein extracted and purified from soybeans, which is a microorganism belonging to the genus Streptomyces and / or Actinoplanes having the ability to produce nociheptide. A method for producing a nociheptide characterized in that: