文献类型：期刊文献

英文题名：Complete genome sequence of high-yield strain S. lincolnensis B48 and identification of crucial mutations contributing to lincomycin overproduction

作者：Wang, Ruida[1];Kong, Fanjing[1];Wu, Haizhen[1,2];Hou, Bingbing[1,2];Kang, Yajing[1];Cao, Yuan[1];Duan, Shiwei[3];Ye, Jiang[1,2];Zhang, Huizhan[1,2]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]East China Univ Sci & Technol, Dept Appl Biol, Shanghai 200237, Peoples R China;[3]Ningbo Univ, Sch Med, Ctr Med Genet, Ningbo 315211, Peoples R China

年份：2020

卷号：5

期号：2

起止页码：37

外文期刊名：SYNTHETIC AND SYSTEMS BIOTECHNOLOGY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000544104800001)】；

基金：This study was supported by the National Natural Science Foundation of China (NSFC) (31900059), the China Postdoctoral Science Foundation (2019M650079), and the Research Program of State Key Laboratory of Bioreactor Engineering. We thank Dr. Weihong Jiang (Institute of Plant Physiology and Ecology, Chinese Academy of Sciences) for kindly providing the plasmid pKCcas9dO.

语种：英文

外文关键词：Lincomycin; Comparative genomics; Regulation; Overproduction

摘要：The lincosamide family antibiotic lincomycin is a widely used antibacterial pharmaceutical generated by Streptomyces lincolnensis, and the high-yield strain B48 produces 2.5 g/L lincomycin, approximately 30-fold as the wild-type strain NRRL 2936. Here, the genome of S. lincolnensis B48 was completely sequenced, revealing a similar to 10.0 Mb single chromosome with 71.03% G + C content. Based on the genomic information, lincomycin-related primary metabolism network was constructed and the secondary metabolic potential was analyzed. In order to dissect the overproduction mechanism, a comparative genomic analysis with NRRL 2936 was performed. Three large deletions (LDI-III), one large inverted duplication (LID), one long inversion and 80 small variations (including 50 single nucleotide variations, 13 insertions and 17 deletions) were found in B48 genome. Then several crucial mutants contributing to higher production phenotype were validated. Deleting of a MarR-type regulator-encoding gene slinc377 from LDI, and the whole 24.7 kb LDII in NRRL 2936 enhanced lincomycin titer by 244% and 284%, respectively. Besides, lincomycin production of NRRL 2936 was increased to 7.7-fold when a 71 kb supercluster BGC33 from LDIII was eliminated. As for the duplication region, overexpression of the cluster situated genes lmbB2 and lmbU, as well as two novel transcriptional regulator-encoding genes (sIinc191 and slinc348) elevated lincomycin titer by 77%, 75%, 114% and 702%, respectively. Furthermore, three negative correlation genes (slinc6156, slinc4481 and sIinc6011) on lincomycin biosynthesis, participating in regulation were found out. And surprisingly, inactivation of RNase J-encoding gene sIinc6156 and TPR (tetratricopeptide repeat) domain-containing protein-encoding gene sIinc4481 achieved lincomycin titer equivalent to 83% and 68% of B48, respectively, to 22.4 and 18.4-fold compared to NRRL 2936. Therefore, the comparative genomics approach combined with confirmatory experiments identified that large fragment deletion, long sequence duplication, along with several mutations of genes, especially regulator genes, are crucial for lincomycin over-production.