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Identification of the Biosynthetic Gene Cluster for the anti-MRSA Lysocins through Gene Cluster Activation Using Strong Promoters of Housekeeping Genes and Production of New Analogs in Lysobacter sp. 3655

文献类型： 外文期刊

作者： Yu, Lingjun 1 ; Du, Fengyu 2 ; Chen, Xusheng 2 ; Zheng, Yongbiao 2 ; Morton, Martha 2 ; Liu, Fengquan 1 ; Du, Liangche 1 ;

作者机构： 1.Jiangsu Acad Agr Sci, Inst Plant Protect, Nanjing 210014, Peoples R China

2.Univ Nebraska, Dept Chem, Lincoln, NE 68588 USA

3.Qingdao Agr Univ, Coll Chem & Pharm, Qingdao 266109, Peoples R China

4.Jiangnan Univ, Sch Biotechnol, Wuxi 214122, Jiangsu, Peoples R China

5.Fujian Normal Univ, Coll Life Sci, Fuzhou 350117, Peoples R China

关键词： Lysobacter; natural products; antibiotics; lysocins; promoter engineering

期刊名称：ACS SYNTHETIC BIOLOGY （ 影响因子：5.11； 五年影响因子：5.239 ）

ISSN： 2161-5063

年卷期： 2020 年 9 卷 8 期

页码：

收录情况： SCI

摘要： The Gram-negative gliding bacteria Lysobacter represent a new and rich source for bioactive natural products. In an effort to discover new antibiotics, we found a cryptic biosynthetic gene cluster (BGC) in Lysobacter sp. 3655 that shared a high similarity with the putative lysocin BGC identified in silico previously from Lysobacter sp. RH2180-S. Lysocins are cyclic lipodepsipeptides with potent activity against MRSA (methicillinresistant Staphylococcus aureus) using a novel mode of action, but the lysocin BGC had not been experimentally verified so far. Using an activity-guided screening, we isolated the main antibiotic compound and confirmed it to be lysocin E. However, the putative lysocin BGC was barely transcribed in the wild type, in which Time lysocins were produced only in specific conditions and in a negligible amount. To activate the putative lysocin BGC, we screened for strongly transcribed housekeeping genes in strain 3655 and found several powerful promoters. Upon engineering the promoters into the BGC, the lysocin gene transcription was significantly enhanced and the lysocin yield was markedly increased. With readily detectable lysocins production in the engineered strains, we showed that lysocin production was abolished in the gene deletion mutant and then restored in the complementary strain, even when grown in conditions that did not support the wild type for lysocin production. Moreover, the engineered strain produced multiple new lysocin congeners. The determination of the lysocin BGC and the Lysobacter promoters will facilitate the ongoing efforts for yield improvement and new antibiotic biosynthesis using synthetic biology strategies.

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