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Enhancement of fengycin production in Bacillus amyloliquefaciens by genome shuffling and relative gene expression analysis using RT-PCR.

Authors
  • Zhao, Junfeng1
  • Zhang, Chong2
  • Lu, Jing2
  • Lu, Zhaoxin2
  • 1 a College of Food Science and Engineering, Henan University of Science and Technology, Tianjing Road, Luoyang 471003, People's Republic of China.
  • 2 b College of Food Science and Technology, Nanjing Agricultural University, Jiangsu, Nanjing 210095, People's Republic of China.
Type
Published Article
Journal
Canadian Journal of Microbiology
Publisher
Canadian Science Publishing
Publication Date
May 2016
Volume
62
Issue
5
Pages
431–436
Identifiers
DOI: 10.1139/cjm-2015-0734
PMID: 27035066
Source
Medline
Keywords
License
Unknown

Abstract

Genome shuffling is an efficient approach for the rapid engineering of microbial strains with desirable industrial phenotypes. In this study, we used genome shuffling in an attempt to improve fengycin production of the wild-type strain Bacillus amyloliquefaciens ES-2-4. After 2 rounds of genome shuffling, a high-yield recombinant F2-72 (FMB72) strain that exhibited 8.30-fold increases in fengycin production was obtained. Comparative analysis of synthetase gene (fenA) expression was conducted between the initial and shuffled strains using fluorescent quantitation RT-PCR. Delta CT (threshold cycle) relative quantitation analysis revealed that fengycin synthetase gene (fenA) expression at the transcriptional level in the FMB72 strain was 12.77-fold greater than in the ES-2-4 wild type. The shuffled strain has a potential application in food and pharmaceutical industries. At the same time, the analysis of improved phenotypes will provide more valuable data for inverse metabolic engineering.

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