title: Enhancement of fengycin production in Bacillus amyloliquefaciens by genome shuffling and relative gene expression analysis using RT-PCR.
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.
authors.affiliation: a College of Food Science and Engineering, Henan University of Science and Technology, Tianjing Road, Luoyang 471003, People's Republic of China.
authors.forename: Junfeng
authors.initials: J
authors.lastname: Zhao
authors.affiliation: b College of Food Science and Technology, Nanjing Agricultural University, Jiangsu, Nanjing 210095, People's Republic of China.
authors.forename: Chong
authors.initials: C
authors.lastname: Zhang
authors.affiliation: b College of Food Science and Technology, Nanjing Agricultural University, Jiangsu, Nanjing 210095, People's Republic of China.
authors.forename: Jing
authors.initials: J
authors.lastname: Lu
authors.affiliation: b College of Food Science and Technology, Nanjing Agricultural University, Jiangsu, Nanjing 210095, People's Republic of China.
authors.forename: Zhaoxin
authors.initials: Z
authors.lastname: Lu
mesh_terms: D000069977:Bacillus amyloliquefaciens / Q000235:genetics / Q000378:metabolism*; D032621:DNA Shuffling; D004269:DNA, Bacterial; D020869:Gene Expression Profiling; D016680:Genome, Bacterial; D055666:Lipopeptides / Q000096:biosynthesis*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D004269:DNA, Bacterial; D055666:Lipopeptides; C049972:fengycin
keywords: Bacillus amyloliquefaciens; CT (cycle-seuil); CT (threshold cycle); RT-PCR; brassage génomique; fengycin; fengycine; genome shuffling