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Streptomycete culture medium and its optimization process and application method

A culture medium, Streptomyces technology, applied in the directions of biochemical equipment and methods, microorganism-based methods, bacteria, etc., can solve the problems of high cost, many by-products, low yield, etc., and achieve lower production costs, reduced by-products, The effect of increased production of antibiotics

Inactive Publication Date: 2006-10-11
EAST CHINA UNIV OF SCI & TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This formula not only has low output, but also has many by-products, and the cost is very high

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] 1. According to glucose 2%, glycerol 1.389%, yeast powder 0.3%, peptone 0.101%, NaCl 1%, FeSO 4 ·7H 2 O 0.05%, CuSO 4 A ratio of 0.00428% is weighed to take an appropriate amount of each component of the medium (except starch and industrial amino acid powder), and add an appropriate amount of deionized water to dissolve.

[0022] 2. Weigh an appropriate amount of starch according to the ratio of 0.2%, add a small amount of amylase, heat to about 80 degrees Celsius, let it stand for 15 minutes, and then heat to boiling. At this time, the solution becomes transparent and slightly fluorescent.

[0023] 3. Weigh an appropriate amount of industrial amino acid powder according to the ratio of 0.5%, and heat it to a solution state. At this time, the color will be slightly darker, but it contains insoluble solids. Vacuum filtration 3 to 4 times to remove impurities.

[0024] 4. Put the above ingredients in the same beaker, add water to make up the volume, and put it into the...

Embodiment 2

[0027] Using the above medium, ferment and culture the following strains: 1) glycosyltransferase gene fscMI interruption mutant strain CS101; 2) cytochrome P450 monooxygenase gene mutant strain CS103; 3) antibiotic FR-008 / Candicidin (candicidin) .

[0028] Medium composition:

[0029] Glucose 2% Starch 1% Glycerin 1.389% Yeast powder 0.3% Industrial amino acid powder 0.5% Peptone 0.101% NaCl 1% FeSO 4 ·7H 2 O 0.05% CuSO 4 0.00428%

[0030] Before sterilization, the pH was adjusted to 6.90 with 5 mol / l NaOH solution.

[0031] Culture method: the inoculation time is 30 hours, the inoculum size is 10%, 28 degree shaker, 200rpm culture, the fermentation level of antibiotics CS101, CS103 and FR-008 / Candicidin are detected in bottles are 216mg / l, 40mg / l and 252mg / l respectively l.

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PUM

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Abstract

This invention relates to a culture media of streptomycete, the process optimization and application thereof. The formula comprises: glucose 1-3%, starch 1-3%, glycerol 1-3%, yeast extract 0.1-0.5%, industrial amino acids powder 0.2-1%, peptone 0.05-0.2%, NaCl 0.5-2%, FeSO4 7H2O 0.01-0.1%, CuSO4 0.002-0.01%, and surplus water. Said culture media can be used in genetic engineering antibiotics fermentation production. It can also satisfy the need of shake flask test, and can be successfully scaled up to fermentator production. Besides, it has few side by-products, which benefits the downstream purification. Compared to the traditional culture media, the yield of the antibiotic FR-008 / Candicidin and its genetic engineering derivatives using this formula is increased by 3-10 times, and the production cost is largely reduced. Furthermore, it is the first time to discover that CuSO4 can dramatically improve the synthesis of antibiotic FR-008 / Candicidin and its genetic engineering derivatives CS101, CS103 and so on.

Description

technical field [0001] The invention relates to a streptomyces culture medium and its optimization process and application method. Background technique [0002] Streptomyces FR-008 is a new strain of antibiotic-producing Streptomyces discovered by Liang Rongfang et al. in the process of studying the protoplast fusion breeding of Nongkang 5192-producing bacteria. Secondly, as for the metabolite FR-008, preliminary chemical structure analysis shows that the chemical structures of FR-008 and Candicidin are the same. The research group led by academician Deng Zixin of Shanghai Jiaotong University has achieved remarkable results in combinatorial biosynthesis, and synthesized a series of genetic engineering derivative compounds of antibiotic FR-008 / Candicidin. These include glycosyltransferase gene fscMI interruption mutant strain CS101, ketose aminotransferase gene fscMII interruption mutant strain CS102 and cytochrome P450 monooxygenase gene mutant strain CS103, which are genet...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P1/04C12R1/465
Inventor 沈亚领魏东芝毛相朝杨亮邓子新陈实
Owner EAST CHINA UNIV OF SCI & TECH