Enhancement of fengycin production in Bacillus amyloliquefaciens by genome shuffling and relative gene expression analysis using RT-PCR
Enhancement of fengycin production in Bacillus amyloliquefaciens by genome shuffling and relative gene expression analysis using RT-PCR
复制标题
通过基因组改组和 RT-PCR 相关基因表达分析增强解淀粉芽孢杆菌中的风霉素产量
DOI：
10.1139/cjm-2015-0734
复制
发表时间：
2016-05-01
影响因子：
2.8
通讯作者：
Lu, Zhaoxin
中科院分区：
文献类型：
--
作者：
Zhao, Junfeng;Zhang, Chong;Lu, Zhaoxin
基因组改组是一种快速工程改造具有理想工业表型的微生物菌株的有效方法。在这项研究中，我们使用基因组改组，试图提高丰霉素生产的野生型菌株解淀粉芽孢杆菌ES-2-4。经过2轮基因组改组，获得了一株高产的重组F2-72（FMB 72）菌株，其产量提高了8.30倍。利用荧光定量RT-PCR对合成酶基因（fenA）的表达进行了比较分析。Δ C-T（阈值循环）相对定量分析显示，FMB 72菌株中芬枯草菌素合成酶基因（fenA）在转录水平上的表达是ES-2-4野生型中的12.77倍。重组菌在食品和医药工业中具有潜在的应用价值。同时，对改良表型的分析将为逆代谢工程提供更有价值的数据。
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 C-T (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.