Culture medium for culturing fusicoccum amygdali del - producer of fusicoccin

FIELD: chemistry.

SUBSTANCE: invention relates to microbological industry, particularly production of biologically active substances and can be used in producing fusicoccins for different purposes, for both agricultural and medical purposes. The culturing medium contains molasses, raw sugar, potassium nitrate (KNO3), potassium phosphate (KH2PO4), magnesium sulphate (MgSO4 x 7H2O) and tap water in a given ratio.

EFFECT: invention increases the range of fusicoccins.

4 ex

 

The invention relates to the microbiological industry, in particular the production of biologically active substances, and can be used to obtain fusicoccin for various purposes as for agricultural production, and for medical purposes.

Currently being actively studied properties and biological activity of fusicoccin in medicine. 1. Sassa T. et al. Fusicoccins P and Q, and 3-epifusicoccins H and Q, new polar fusicoccins isolate from Niigata 2-A of a peach Fusicoccum canker fungus. Biosci. Biotechnol Biochem. 2002, Nov; 66(11), 2356-61. 2. Ingrid J. de Vries - van Leeuwen et al. Fusicoccin A - selectively dosage apoptosis in tumor cells after interferon-alpha priming. Cancer Lett. 2010, 293(2), 198-206.

Known nutrient medium for culturing producer fusicoccin (copyright certificate №1172515, CL A01N 63/00, 07.07.1980), including glucose, soy flour, disubstituted potassium phosphate, potassium sulfate and magnesium sulfate and water. The lack of nutrient - receiving mainly fusicoccin A.

Also known nutrient medium for culturing producer fusicoccin (copyright certificate №1378364, CL C12N 1/14, A01N 63/00, 04.12.1984), including molasses, glucose, potassium nitrate one-deputizing, magnesium phosphate, magnesium sulfate and water. The lack of this nutrient medium - high levels in the spectrum of fusicoccin of fusicoccin A.

The task of the invention is to obtain the greatest SP the CTE of fusicoccin to use them for medical purposes.

The technical result is to obtain in addition to fusicoccin And fusicoccin C, D, H, J.

This object is achieved due to the fact that nutrient media for cultivation of Fusicoccum amygdali Del. producer fusicoccin, including, wt.%: molasses 1.0-2.0; KNO30.02-0.03; KH2PO40.1-0.3; water - up to 100, further comprises raw sugar 0.8-1.5; MgSO4×7H2O 0.02-0.03.

The specified interval, the content of components in the environment are sufficient to achieve the objectives. The increase in the interval leads to unnecessary consumption of raw materials. Reducing the quantitative values of the components does not allow to achieve the project objectives due to the fact that does not provide the required level of biosynthesis of fusicoccin.

Examples of specific performance.

Example 1. Prepare the medium composition, wt.%: molasses 2.0; raw sugar 1.5; KNO30.03; KH2PO40.3; MgSO4×7H2O 0.03; tap water to 100; pH 6.0-6.5. Biosynthesis is carried out on two-phase system. The specified medium inoculated with the fungus Fusicoccum amygdali Del. and grow the seed mycelium within 48 hours. Then take 10-15 ml of seed mycelium and inoculated in flasks with 150 ml of culture medium of the same composition and cultured at a temperature of 25-26°C for 72 hours. Get the culture fluid with the concentration of fusicoccin 500 mg/L. Using the combined method of gas chromatography - mA is with spectrometry (GC-MS) has allowed to establish, what in the culture fluid contents of fusicoccin varies within the following limits: fusicoccin C 28-21%, D is 2,5-1,5%, H 8,0-6,0%, J 20-15%, A 40-49%, and 16-O-dimethylpiperazine J, directantiglobulin and separate penderitaan fusicoccin metabolites.

Example 2. Prepare the medium composition, wt.%: molasses 1.0; raw sugar 0.8; KNO30.02; KH2PO40.1; MgSO4×7H2O 0.02; tap water to 100; pH 6.0-6.5. The environment is seeded with a culture of the fungus as described in example 1, were cultured under the same conditions. The concentration of fusicoccin in culture medium 500 mg/l In the culture fluid contents of fusicoccin varies within the following limits: fusicoccin C 28-21%, D 2.5-1.5%, H 8.0-6.0%, J 20-15%, A 40-49%, a 16-O-dimethylpiperazine J, directantiglobulin and separate penderitaan fusicoccin metabolites.

Example 3. Prepare the environment, including, wt.%: molasses 1.5; raw sugar 1.2; KNO30.025; KH2PO40.2; MgSO4×7H2O 0.025; tap water to 100; pH 6.0-6.5. The environment is seeded with a culture of the fungus as described in example 1, were cultured under the same conditions. The concentration of fusicoccin in culture medium 500 mg/l In the culture fluid contents of fusicoccin varies within the following limits: fusicoccin C 28-21%, D 2.5-1.5%, H 8.0-6.0%, J 20-15%, A 40-49%, a 16-O-dimethylpiperazine J, directantiglobulin and pindaric avannah fusicoccin metabolites.

Example 4. Prepare the environment, including, wt.%: molasses 0.5; raw sugar 0.4; KNO30.01; KH2PO40.05; MgSO4×7H2O 0,01; tap water to 100; pH 6.0-6.5. The environment is seeded with a culture of the fungus as described in example 1, were cultured under the same conditions. The concentration of fusicoccin in culture liquid is reduced to 280 mg/l In the culture fluid content increases fusicoccin but the content of other fusicoccin decreases - C 11%D 1%H 2%, J 6%, A 68%, and 16-O-dimethylpiperazine J, directantiglobulin and separate penderitaan fusicoccin metabolites.

Conclusion. The alleged invention allows to increase sufficiently the range of fusicoccin for medical purposes.

Nutrient media for cultivation of Fusicoccum amygdali Del. producer fusicoccin, including carbohydrates, potassium phosphate, magnesium sulfate and water, characterized in that the medium additionally contains potassium nitrate and carbohydrate - raw sugar and molasses in the following ratio, wt.%:

molasses1,0-2,0
raw sugar0,8-1,5
KNO30,02-0,03
KH2PO4 of 0.1-0.3
MgSO4·7H2O0,02-0,03
water water100



 

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