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Method for increasing yield of antibacterial peptide of bacillus subtilis through overexpression of yerP gene

A Bacillus subtilis, overexpression technology for use in microorganism-based methods, biochemical equipment and methods, bacteria, etc.

Inactive Publication Date: 2011-10-19
NANJING AGRICULTURAL UNIVERSITY
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Problems solved by technology

[0004] At present, in the world, the methods to increase its production are mainly to screen excellent strains and optimize fermentation conditions, but few use molecular biology techniques to improve strains method to increase the production of antimicrobial substances

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  • Method for increasing yield of antibacterial peptide of bacillus subtilis through overexpression of yerP gene
  • Method for increasing yield of antibacterial peptide of bacillus subtilis through overexpression of yerP gene
  • Method for increasing yield of antibacterial peptide of bacillus subtilis through overexpression of yerP gene

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specific Embodiment approach

[0038] The present invention first constructs yerP Gene overexpression vector pHCMC-YerP, after electroporation, the pHCMC-YerP vector was transformed into Bacillus subtilis ( Bacillus subtilis ) ATCC 9943 (purchased from The Global Bioresource Center), which was transformed into Bacillus subtilis ATCC 9943 by electroporation to achieve overexpression yerP The purpose of this gene is to construct a Bacillus subtilis strain FMB 45 with high production of antimicrobial peptides. The mutant strains were identified by extracting plasmids; the yields of surfactin and fengycin in the improved strains were analyzed by high performance liquid chromatography (HPLC). The specific implementation is as follows:

[0039] (1) yerP Construction of Gene Overexpression Vector pHCMC-YerP

[0040] Bacillus subtilis P43 Promoter gene design primers P43-F and P43-R to Bacillus subtilis ( Bacillus subtilis ) ATCC 9943 genomic DNA was used as a template to amplify the P43 promoter gene by ...

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Abstract

The invention relates to a method for increasing the yield of antibacterial peptide of bacillus subtilis through the overexpression of yerP gene, and belongs to the field of biotechnology. The method comprises the following steps of: constructing a gene overexpression vector pHCMC-YerP by adopting bacillus subtilis ATCC9943 as initial bacterial strain and escherichia coli cloning vector pHCMC04 as a framework, delivering the gene overexpression vector pHCMC-YerP into bacillus subtilis ATCC9943 through electrotransformation to achieve the overexpression of yerP gene, thereby constructing a high-yield antibacterial-peptide bacterial strain FMB45. Verified by high-performance liquid chromatography, the surfactin and fengycin producing capability of the improved bacterial strain is greatly improved.

Description

1. Technical field [0001] The present invention relates to the overexpression of yerP The invention relates to a method for improving the production of antimicrobial peptides of Bacillus subtilis by gene, which belongs to the field of biotechnology. 2. Background technology [0002] Bacillus subtilis ( Bacillus subtilis ) is currently recognized as one of the safe microorganisms, and its antibacterial metabolites are rich and diverse, especially lipopeptide antibacterial substances, which not only have a wide antibacterial spectrum but also have the function of biosurfactant, so they can not only be used for biological control in agriculture , and also has a wide range of applications in industry and environmental protection. [0003] Bacillus subtilis ( Bacillus subtilis ) ATCC 9943 is a Bacillus subtilis that can secrete surfactin, fengycin and other antimicrobial peptide substances (Valérie Leclère, Romain Marti, Max Béchet. [0004] At present, in the world, the me...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/75C12N1/21C12P21/02C12R1/125
Inventor 陆兆新曹国强吕凤霞别小妹张充钟蕾
Owner NANJING AGRICULTURAL UNIVERSITY
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