Genome shuffling of Bacillus amyloliquefaciens for improving antimicrobial lipopeptide production and an analysis of relative gene expression using FQ RT-PCR.

Journal of Industrial Microbiology & Biotechnology
Junfeng ZhaoZhaoxin Lu

Abstract

Genome shuffling is an efficient approach for the rapid improvement of the yield of secondary metabolites. This study was undertaken to enhance the yield of surfactin produced by Bacillus amyloliquefaciens ES-2-4 using genome shuffling and to examine changes in SrfA expression of the improved phenotype at the transcriptional level. Six strains with subtle improvements in lipopeptide yield were obtained from populations generated by ultraviolet irradiation, nitrosoguanidine, and ion beam mutagenesis. These strains were then subjected to recursive protoplast fusion. A strain library that was likely to yield positive colonies was created by fusing the lethal protoplasts obtained from both ultraviolet irradiation and heat treatments. After two rounds of genome shuffling, a high-yield recombinant F2-38 strain that exhibited 3.5- and 10.3-fold increases in surfactin production in shake flask and fermenter respectively, was obtained. Comparative analysis of synthetase gene expression was conducted between the initial and shuffled strains using FQ (fluorescent quantitation) RT-PCR. Delta CT (threshold cycle) relative quantitation analysis revealed that surfactin synthetase gene (srfA) expression at the transcriptional level in the F2-3...Continue Reading

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Citations

May 10, 2013·Journal of Industrial Microbiology & Biotechnology·Ji ZhangWensheng Xiang
May 17, 2013·Journal of Industrial Microbiology & Biotechnology·Pu ZhengZhihao Sun
Jan 16, 2014·Applied Biochemistry and Biotechnology·Guanghai YuJiang Zhao
Feb 18, 2014·International Journal of Biological Macromolecules·Bo YuanJi-Hong Jiang
Mar 7, 2014·Applied Microbiology and Biotechnology·Damien Biot-Pelletier, Vincent J J Martin
Aug 8, 2014·Applied Biochemistry and Biotechnology·Yuping ZhaoJingzhu Yang
Jul 21, 2015·Peptides·Mnif Inès, Ghribi Dhouha
Jun 5, 2015·World Journal of Microbiology & Biotechnology·Seema PatelJ Satya Eswari
Jul 6, 2014·Current Opinion in Biotechnology·Robin GeysInge Van Bogaert
Oct 16, 2016·Bioprocess and Biosystems Engineering·Philippe JacquesRénato Froidevaux
Jun 15, 2018·Brazilian Journal of Microbiology : [publication of the Brazilian Society for Microbiology]·Junfeng ZhaoZhaoxin Lu
Feb 28, 2020·International Journal of Environmental Research and Public Health·Deepti MalviyaG P Brahmaprakash

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