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Enhanced production of iturin A in Bacillus amyloliquefaciens by genetic engineering and medium optimization
Process Biochemistry ( IF 4.4 ) Pub Date : 2020-03-01 , DOI: 10.1016/j.procbio.2019.11.017
Yuxiang Xu , Dongbo Cai , Hong Zhang , Lin Gao , Yong Yang , Jiaming Gao , Yanyan Li , Chunlei Yang , Zhixia Ji , Jun Yu , Shouwen Chen

Abstract Brown spot disease, caused by Alternaria alternate, is one of the most destructive leaf spot diseases that made a major impact on tobacco growing. Biocontrol has attracted increasing attentions as its features of environmental friendship, promoting plant growth, etc. Iturin A, with broad spectrum antifungal activity, is a kind of biosurfactant that mainly produced by Bacillus. In this study, the promoter of iturin A synthetase cluster of Bacillus amyloliquefaciens HZ-12 was respectively replaced by promoters P43, PbacA, PsrfA and Pylb, our results implied that transcriptional level of gene ituD and iturin A titer showed consistent change trends, and PbacA was proven as the most efficient promoter, iturin A titer of which reached 950.08 ± 19.43 mg/L. Furthermore, regulator gene abrB was deleted to release the repression effect of AbrB on PbacA, ituD transcriptional level and iturin A titer were increased by 133.25 % and 20.88 %, respectively. Then, the maximum iturin A titer reached 2013.43 ± 32.86 mg/L by optimizing fermentation medium, increased by 392.15 % compared to the original (408.97 ± 21.35 mg/L). Finally, our results demonstrated that enhancing iturin A synthetic capability benefited the suppression of A. alternate. Collectively, this study provided a promising strain with an efficient fermentation medium for large-scale industrial production of iturin A.

中文翻译:

通过基因工程和培养基优化提高解淀粉芽孢杆菌中 iturin A 的产量

摘要 褐斑病是由交替链格孢引起的,是最具破坏性的叶斑病之一,对烟草种植产生了重大影响。生物防治因其对环境友好、促进植物生长等特点越来越受到人们的关注。Iturin A具有广谱抗真菌活性,是一种主要由芽孢杆菌产生的生物表面活性剂。本研究中解淀粉芽孢杆菌HZ-12的iturin A合成酶簇的启动子分别被启动子P43、PbacA、PsrfA和Pylb取代,结果表明ituD基因的转录水平和iturin A的滴度呈现一致的变化趋势,PbacA被证明是最有效的启动子,其中 iturin A 滴度达到 950.08 ± 19.43 mg/L。此外,删除调节基因 abrB 以释放 AbrB 对 PbacA 的抑制作用,ituD 转录水平和 iturin A 滴度分别增加了 133.25% 和 20.88%。然后,通过优化发酵培养基,最大iturin A滴度达到2013.43±32.86mg/L,比原来的(408.97±21.35mg/L)提高了392.15%。最后,我们的结果表明,增强 iturin A 的合成能力有利于抑制 A. alternation。总的来说,这项研究为伊图林 A 的大规模工业生产提供了一种具有高效发酵培养基的有前景的菌株。与原始浓度 (408.97 ± 21.35 mg/L) 相比增加了 15 %。最后,我们的结果表明,增强 iturin A 的合成能力有利于抑制 A. alternation。总的来说,这项研究为伊图林 A 的大规模工业生产提供了一种具有高效发酵培养基的有前景的菌株。与原始浓度 (408.97 ± 21.35 mg/L) 相比增加了 15 %。最后,我们的结果表明,增强 iturin A 的合成能力有利于抑制 A. alternation。总的来说,这项研究为伊图林 A 的大规模工业生产提供了一种具有高效发酵培养基的有前景的菌株。
更新日期:2020-03-01
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