Streptomyces aureus mutant strain with high aureomycin yield and low impurity proportion and application thereof

A technology of Streptomyces aureus and chlortetracycline, which is applied in the biological field, can solve the problems of saturation effect of mutagenesis, difficulty in increasing yield, low mutagenesis efficiency, etc., and achieves the goal of promoting the synthesis of chlortetracycline, reducing enterprise cost, and strengthening seed culture. Effect

Active Publication Date: 2021-10-26
金河生物科技股份有限公司 +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the industrial strains of Streptomyces aureus generally undergo multiple traditional mutagenesis, often appearing in the phenomenon of low mutagenesis efficiency,

Method used

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  • Streptomyces aureus mutant strain with high aureomycin yield and low impurity proportion and application thereof
  • Streptomyces aureus mutant strain with high aureomycin yield and low impurity proportion and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] The breeding of embodiment 1 Streptomyces aureus FJC-505

[0069] On the basis of the starting strain Streptomyces aureus D29 strain, the method of protoplast compound mutagenesis is used to improve the output of aureomycin, reduce impurities, and select and breed excellent strains with stable genetics. The specific methods are as follows:

[0070] 1. Collection of starting strain mycelium

[0071] Inoculate the glycerol bacteria of the starting bacterial strain Streptomyces aureus D29 bacterial strain to solid slant medium (18 * 180mm test tube slant, wherein the preparation method of slant and solid plate medium is as follows: bran 40.0g, potassium dihydrogen phosphate 0.2g, magnesium sulfate 0.2g, 0.5g diammonium hydrogen phosphate, 20.0g agar, distilled water to 1000mL; natural pH, 121°C high-pressure steam sterilization for 20-30min), after inoculation, the bacteria were cultured at 34°C and humidity 45% for 4 days , and then use 5mL sterile normal saline to elute...

Embodiment 2

[0097] Embodiment 2: Breeding of Streptomyces aureus FJW-507

[0098] On the basis of the starting strain Streptomyces aureus D29 strain, the method of protoplast compound mutagenesis is used to improve the output of aureomycin, reduce impurities, and select and breed excellent strains with stable genetics. The specific methods are as follows:

[0099] Steps 1, 2, 3, 4, 5, 6 are the same as the breeding steps 1, 2, 3, 4, 5, 6 of Streptomyces aureus FJC-505 in Example 1.

[0100] Wherein, bacterial strain Streptomyces aureus FJW-507 passage fermentation experiment result is as follows:

[0101] The impact of table 2 passage on Streptomyces aureus FJW-507 aureomycin content and impurity

[0102]

[0103] The results showed that the five generations of Streptomyces aureus FJW-507 had no significant effect on the fermentation level, the contents of aureomycin, tetracycline and desmethylaureomycin in the fermentation liquid did not change significantly during the subculture pro...

Embodiment 3

[0106] Embodiment 3: the method for Streptomyces aureus FJC-505 and Streptomyces aureus FJW-507 fermentation strengthening

[0107] First, prepare a seed strengthening medium (spore medium): weigh 30 g of bran, mix it with 30 ml of nutrient solution, and steam for 30 minutes to obtain a bran nutrient solution medium. Weigh 30 g of bran nutrient solution and culture it in an eggplant bottle, and perform high-temperature and high-pressure sterilization according to conventional methods, at 121° C. for 20-30 min. The nutrient solution preparation method is as follows: 30.0g cornstarch, 2.0g peanut cake powder, 5.0g yeast powder, 3.0g peptone, 2.0g ammonium chloride, 2.0g sodium chloride, 0.1g potassium dihydrogen phosphate, 0.15g magnesium chloride, Magnesium sulfate 0.15g, distilled water to 1000ml. This is the bran seed strengthening medium (spore medium).

[0108] Then, prepare Streptomyces aureus FJC-505 and Streptomyces aureus FJW-507 spore suspension, and get 2.5mL spore ...

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Abstract

The invention discloses a streptomyces aureus mutant strain, a breeding method thereof and an optimized culture medium. The mutant strain can efficiently synthesize and accumulate aureomycin, the content of impurity tetracycline and demeclocycline is remarkably reduced, the genetic stability is good, the form, color, aureomycin production level and main impurity content of the mutant strain are basically stable after five generations of continuous passage, and the mutant strain can be applied to industrial fermentation production and can be used as a production strain for further research and development. The optimized culture medium is beneficial to promoting synthesis and accumulation of the aureomycin, and the proportion of the impurity tetracycline and demeclocycline is further reduced. A high-yield strain breeding model is constructed based on multiple compound mutagenesis and precursor/product tolerance, the diversity of a mutation library is higher, simpleness, convenience and quickness are achieved, the screening efficiency is improved, the screening workload is reduced, and certain application value and guiding significance for reducing the production cost and improving the productivity of the aureomycin and other streptomycete source antibiotics are achieved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a Streptomyces aureus mutant strain with high aureomycin yield and low impurity ratio and application thereof. Background technique [0002] Chlortetracycline is a tetracycline broad-spectrum antibiotic produced by the fermentation of Streptomyces aureofaciens. It has the characteristics of antibacterial, growth-promoting, high feed utilization rate, low residue in the body, and its production technology is mature and The advantage of low production cost has become the most widely used antibacterial and growth-promoting agent in the feed industry. As an important medicinal microorganism, Streptomyces aureus is a highly aerobic, Gram-positive actinomycete. However, currently, the titer of the bacterial strains fermenting and producing chlortetracycline is not high, and the production stability is not good, which seriously affects the fermentation index and production effi...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/38C12N15/01C12N13/00C12P29/00C12R1/49
CPCC12N1/20C12N1/38C12N15/01C12N13/00C12P29/00
Inventor 谢昌贤王月清蔡玉凤梁玲黄钦耿赵燕玉翁雪清陈健孙岩宁陈红梅王鹏飞谢必峰黄建忠
Owner 金河生物科技股份有限公司
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