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Method for producing aureomycin by using streptomyces aureus

A technology of Streptomyces aureus and aureomycin, which is applied in the biological field and can solve the problems of aging mycelia and affecting the production of aureomycin

Active Publication Date: 2021-09-14
金河生物科技股份有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If the culture conditions are not suitable, it will often cause the mycelium to become old, which will affect the production of aureomycin to a large extent.

Method used

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  • Method for producing aureomycin by using streptomyces aureus
  • Method for producing aureomycin by using streptomyces aureus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] The breeding of embodiment 1 Streptomyces aureus FJC-505

[0081] 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:

[0082] 1. Collection of starting strain mycelium

[0083] Inoculate the glycerol bacteria of the starting bacterial strain Streptomyces aureus D29 strain to a solid slant medium (18 × 180mm test tube slant, wherein the preparation method of the 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℃ high pressure steam sterilization for 20-30min), after inoculation the bacteria were cultured at 34℃, humidity 45% for 4 days , and then use 5mL sterile normal saline to elute the spore...

Embodiment 2

[0109] Embodiment 2: Breeding of Streptomyces aureus FJW-507

[0110] 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:

[0111] Steps 1, 2, 3, 4, 5, and 6 are the same as the breeding steps 1, 2, 3, 4, 5, and 6 of Streptomyces aureus FJC-505 in Example 1.

[0112] Wherein, bacterial strain Streptomyces aureus FJW-507 passage fermentation experiment result is as follows:

[0113] The impact of table 2 passage on Streptomyces aureus FJW-507 aureomycin content and impurity

[0114]

[0115] 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 subcul...

Embodiment 3

[0118] Embodiment 3: the method for Streptomyces aureus FJC-505 and Streptomyces aureus FJW-507 fermentation strengthening

[0119] 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).

[0120] Then, prepare Streptomyces aureus FJC-505 and Streptomyces aureus FJW-507 spore suspensions, and transfer 2.5 mL...

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Abstract

The invention discloses a method for producing aureomycin by using streptomyces aureus. According to the method, a culture medium and a culture method are combined and optimized, so that the synthesis and accumulation of aureomycin are promoted, and the proportion of impurity tetracycline and demethyl aureomycin is further reduced. A high-yield strain breeding model is constructed based on multiple composite mutagenesis and precursor / product tolerance, so that the diversity of a mutation library is higher, the method is simple, convenient and rapid, the screening efficiency is improved, the screening workload is reduced, and the method has certain application value and guiding significance for the reduction of the production cost and the improvement of the productivity of aureomycin andother streptomyces-derived antibiotics. The mutant strain can efficiently synthesize and accumulate aureomycin, the content of the impurity tetracycline and demethyl aureomycin is remarkably reduced, the genetic stability is good, the form, color, aureomycin production level and the content of main impurities are basically stable after continuous passage for five generations, and the mutant strain can be applied to industrial fermentation production and can be used as a production strain for further research and development.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for producing aureomycin by using Streptomycesaureofaciens. Background technique [0002] Chlortetracycline is a broad-spectrum tetracycline antibiotic produced by the fermentation of Streptomyces aureofaciens. As an important medicinal microorganism, Streptomyces aureus is a highly aerobic, Gram-positive actinomycete. If the culture conditions are not suitable, the hyphae will often become old, which will affect the output of aureomycin to a large extent. In order to establish an optimized, economical and competitive biological production process of chlortetracycline, a series of issues such as production level, fermentation method, post-processing and refining must be fully considered. [0003] Microbial fermentation is a complex biological process, which is affected by many factors, among which the composition and ratio of the medium is one of the key factor...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P29/00C12R1/49
CPCC12N1/20C12P29/00
Inventor 谢昌贤王月清赵燕玉蔡玉凤梁玲黄钦耿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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