CN110317841B

CN110317841B - Fermentation production culture medium for griseofulvin

Info

Publication number: CN110317841B
Application number: CN201910648868.5A
Authority: CN
Inventors: 胡志远; 杨春艳; 于彩岩; 安丽艳
Original assignee: Inner Mongolia Glint Pharmaceutical Co ltd
Current assignee: Inner Mongolia Glint Pharmaceutical Co ltd
Priority date: 2019-07-10
Filing date: 2019-07-10
Publication date: 2022-11-22
Anticipated expiration: 2039-07-10
Also published as: CN110317841A

Abstract

The invention discloses a culture medium for fermentation production of griseofulvin, which is prepared from the following raw materials in percentage by weight: 6% of indica rice flour, 5% of corn flour, 3% of sweet potato flour, 0.6% of potassium dihydrogen phosphate, 0.4% of calcium carbonate, 0.1% of sodium nitrate, 0.1% of ammonium sulfate, 0.1% of ferrous sulfate, 0.8% of sodium chloride, 0.7% of potassium chloride, 0.1% of magnesium sulfate, 0.05% of a GPE (meglumine) type, and the balance of water. The viscosity of the fermentation liquor is reduced, the fermentation unit is stabilized at 28000-30000 mu g/ml, the fermentation period is shortened by about 45 hours, the fermentation energy consumption is reduced, the cost is reduced, and the yield is high.

Description

Fermentation production culture medium for griseofulvin

The invention relates to a preparation method of bulk drugs, in particular to a culture medium for fermentation production of griseofulvin.

Griseofulvin is an antibiotic raw material drug with the English name Griseofulvin, the Chinese name Griseofulvin, the microcrystalline Griseofulvin, the English name Fulcin, fulcine, fulvicin, fungivin, grifukulvin, grisivin, griseofulvin, [ CAS ] 126-07-8, and the product is white or white-like fine powder; the product has good antibacterial effect on superficial fungi such as trichophyton, microsporum, epidermophyton, etc.

Griseofulvin is suitable for treating various tinea diseases including tinea capitis, tinea barbae, tinea corporis, tinea cruris, tinea pedis and tinea unguium.

In the existing method for preparing griseofulvin, a, fermentation culture of griseofulvin is carried out for a certain period of heating inactivation, solid-liquid separation of a plate frame, granulation and drying of filter cake mycelia. b. Extracting with acetone to obtain griseofulvin acetone extractive solution; c. decoloring the griseofulvin acetone extract by using activated carbon, and concentrating and crystallizing the decolored solution to obtain a griseofulvin crude product; d. dissolving the griseofulvin crude product by using acetone, decoloring the griseofulvin acetone dissolved solution by using activated carbon, and concentrating and crystallizing the decolored solution to obtain a griseofulvin recrystallization; e. washing griseofulvin recrystallization with butanol, washing with ethanol, drying, and micronizing to obtain griseofulvin raw material.

The invention aims to provide a griseofulvin fermentation production culture medium, which has the advantages of low energy consumption, cost reduction, short fermentation period, fermentation unit improvement and production efficiency improvement in the production process.

The technical scheme adopted by the invention is as follows:

a culture medium for fermentation production of griseofulvin is prepared from the following raw materials in percentage by weight: 6% of long-shaped rice flour, 5% of corn flour, 3% of sweet potato flour, 0.6% of potassium dihydrogen phosphate, 0.4% of calcium carbonate, 0.1% of sodium nitrate, 0.1% of ammonium sulfate, 0.1% of ferrous sulfate, 0.8% of sodium chloride, 0.7% of potassium chloride, 0.1% of magnesium sulfate, 0.05% of kyoto GPE (glutathione) type, and the balance of water.

The invention has the advantages that: 1. the carbon source components and the mixture ratio in the fermentation medium are adjusted, and the production process monitoring data show that the mixture ratio of the carbon source is 6% of long-shaped rice flour, 5% of corn flour and 3% of sweet potato flour, so that the viscosity of the fermentation liquid can be reduced, the fermentation unit is stabilized at 28000-30000 mu g/ml, the fermentation period is shortened by about 45 hours, the fermentation energy consumption is reduced, and the cost is reduced;

2. compared with a comparison formula, the potassium dihydrogen phosphate is adjusted to be 0.6 percent, the calcium carbonate is adjusted to be 0.4 percent, the pH value of the fermentation liquor is reduced from 6.8 to 5.8, and the growth speed of the bacterial strain in the fermentation liquor is obviously increased compared with that of the bacterial strain in the comparison test formula by sampling and observing under a microscope;

3. the addition of a proper amount of GPE (glutathione E) type foam can improve the gas-liquid mixing effect in the fermentation tank, increase the oxygen rate in the fermentation tank, and facilitate the growth of strains and increase the yield;

4. the test formula is applied to industrial mass production, and the yield is about 16 percent higher than that of a reference test formula.

The present invention will be further described below.

The method for producing griseofulvin by using the culture medium comprises the following steps:

a. the culture medium is configured and then is put into a fermentation tank at 121-126 ℃, the sterilization is maintained for 30 minutes, when the temperature of the tank is reduced to 30 ℃, the inoculation and the fermentation are carried out, and the culture conditions are as follows: initial medium pH =5.8, culture temperature: 29 ℃. + -. 1 ℃, air flow: 2500-3600 m ³ H, pot pressure: 0.01-0.05 MPa, supplementing material in the middle stage of fermentation, culturing for 340 h, heating and inactivating, and filtering the strain with plate-and-frame filter pressSeparating mycelium from fermentation broth, adding calcium oxide into mycelium, granulating, drying, and extracting with acetone;

b. extracting with acetone to obtain griseofulvin acetone extractive solution;

c. decoloring the griseofulvin acetone extract by using activated carbon, and concentrating and crystallizing the decolored solution to obtain a griseofulvin crude product;

d. dissolving the griseofulvin crude product with acetone, decoloring the griseofulvin acetone solution with activated carbon, and concentrating and crystallizing the decolored solution to obtain griseofulvin recrystallization;

e. washing griseofulvin recrystallization with butanol, washing with ethanol, drying, and micronizing to obtain griseofulvin raw material.

Comparative experiment:

test formulations (culture Medium used in the invention)

6% of indica rice flour, 5% of corn flour, 3% of sweet potato flour, 0.6% of potassium dihydrogen phosphate, 0.4% of calcium carbonate, 0.1% of sodium nitrate, 0.1% of ammonium sulfate, 0.1% of ferrous sulfate, 0.8% of sodium chloride, 0.7% of potassium chloride, 0.1% of magnesium sulfate, 0.05% of a GPE (meglumine) type, and the balance of water.

Comparative formulation (Medium previously used by this company)

15% of indica rice flour, 0.4% of potassium dihydrogen phosphate, 0.6% of calcium carbonate, 0.1% of sodium nitrate, 0.1% of ammonium sulfate, 0.1% of ferrous sulfate, 0.8% of sodium chloride, 0.7% of potassium chloride, 0.1% of magnesium sulfate and the balance of water.

Mixing the above two formulas, sterilizing, culturing seed culture fluid in a fermentation tank, culturing after middle-stage feeding, inactivating, separating mycelium from fermentation broth, granulating, extracting, decolorizing, crystallizing, washing, drying, and micronizing to obtain griseofulvin raw material. The results are compared by three tests as shown in the table:

table 1 comparative test table

And (4) test conclusion: compared with a comparative formula, the experimental formula has the advantages that the filtration rate of the fermentation liquor in the experimental formula is obviously improved, the filtration yield is about 16 percent higher than that of the comparative formula, the pH value after the fermentation is reduced from 6.8 to 5.8, the fermentation titer is stabilized at 28000-30000 mu g/ml, the fermentation period is shortened by about 45 hours, 150 ten thousand yuan is saved every year, the efficiency and the yield of an enterprise are improved, and the economic benefit is about 1000 ten thousand.

Claims

1. The fermentation production culture medium for griseofulvin is characterized by being prepared from the following raw materials in percentage by weight: 6% of long-shaped rice flour, 5% of corn flour, 3% of sweet potato flour, 0.6% of potassium dihydrogen phosphate, 0.4% of calcium carbonate, 0.1% of sodium nitrate, 0.1% of ammonium sulfate, 0.1% of ferrous sulfate, 0.8% of sodium chloride, 0.7% of potassium chloride, 0.1% of magnesium sulfate, 0.05% of kyoto GPE (glutathione) type, and the balance of water.