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Streptomyces roseospora l31 with high daptomycin production and its construction method

A technology of Streptomyces roseospora and daptomycin, which is applied in the field of Streptomyces roseospora L31 with high yield of daptomycin and its construction field, which can solve the problems of unsatisfactory industrial production and clinical application, low fermentation unit of daptomycin, etc. problems, to achieve the effect of increasing supply, increasing production and speeding up

Active Publication Date: 2021-03-26
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, at present, the fermentation unit of daptomycin is low, which cannot meet the needs of industrial production and clinical application.

Method used

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  • Streptomyces roseospora l31 with high daptomycin production and its construction method

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Experimental program
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Effect test

Embodiment 1

[0014] The culture medium used in this embodiment:

[0015] 1. R5 solid medium: 10.3% sucrose, 1% glucose, 0.5% yeast extract, 0.01% casein hydrolyzate, K 2 SO 4 0.025%, MgCl 2 ·6H 2 O 1.012%, trace element solution 0.2%, trimethylolmethylaminoethanesulfonic acid 0.573%, proline 0.03%, agar powder 2.2%. After sterilization, add 5% KH to every 1L medium 2 PO 4 1mL, 5MCaCl 2 4mL, 700μL of 10M NaOH, the rest is water, the stated percentages are mass percentages, and the pH is natural

[0016] Trace element solution: CuCl 2 2H 2 O 0.1g, Na 2 B 4 o 7 10H 2 O 0.1g, MnCl 2 4H 2 O 0.1g, ZnCl 2 0.4g, (NH4)Mo 7 o 24 4H 2 O 0.1g, FeCl 3 ·6H 2 O 2g, add water to make up to 1L

[0017] 2. TSB medium: 2% of tryptone soybean broth, 5% of polyethylene glycol (PEG600), and the rest is water, the percentages are all by mass, and the pH is natural.

[0018] 3. YEME fermentation medium: 0.3% of yeast extract, 0.5% of peptone, 0.3% of malt extract, 4% of glucose, and the r...

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Abstract

The present invention relates to a Streptomyces roseosporus (Streptomyces roseosporus) L31 with high yield of daptomycin, which fuses two enzymes (DptE and DptF) in the initial reaction of the synthesis pathway of daptomycin, and directional transforms tryptophan The production of daptomycin in this strain was increased by 140% compared with the original strain (WT).

Description

technical field [0001] The invention relates to the field of microbial pharmacy, in particular to a high-daptomycin-producing Streptomyces roseospora L31 and a construction method thereof. Background technique [0002] Due to the widespread use of antibiotics in clinical practice, the drug resistance of pathogenic bacteria has become more and more serious. Humans urgently need new antibiotics to combat the emergence of drug-resistant bacteria. Daptomycin is a cyclolipopeptide antibiotic extracted from the fermentation broth of Streptomyces roseosporus, which has strong antibacterial activity against Gram-positive bacteria and is considered to be the best substitute for vancomycin . Daptomycin binds to the cell membrane of pathogenic bacteria, causing ion outflow and causing their death. Because of its unique cyclolipopeptide structure and mechanism of action, and its lack of cross-resistance with other antibiotics, daptomycin has broad application prospects. [0003] In S...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/21C12P21/04C12R1/465
CPCC07K7/08C12P21/02C12N1/205C12R2001/465
Inventor 李永泉毛旭明袁鹏辉
Owner ZHEJIANG UNIV
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