title: DepR1, a TetR Family Transcriptional Regulator, Positively Regulates Daptomycin Production in an Industrial Producer, Streptomyces roseosporus SW0702.
abstract: Daptomycin is a potent cyclic lipopeptide antibiotic. It is widely used against various Gram-positive bacterial pathogens. Historically, a poor understanding of the transcriptional regulation of daptomycin biosynthesis has limited the options for targeted genetic engineering toward titer improvement. Here, we isolated a TetR family transcriptional regulator, DepR1, from the industrial producer Streptomyces roseosporus SW0702 using a biotinylated dptE promoter (dptEp) as a probe. The direct interaction between DepR1 and dptEp then was confirmed by electrophoretic mobility shift assays and DNase I footprinting assays. The deletion of depR1 led to a reduction in dptEp activity and the cessation of daptomycin production. The ΔdepR1 mutant produced less red pigment and failed to sporulate on R5 medium. This suggests that DepR1 plays a positive role in the control of morphological differentiation. Moreover, DepR1 was positively autoregulated by directly binding to its own promoter. This might account for the positive feedback regulation of daptomycin production. Based on these positive effects, genetic engineering by overexpression of depR1 raised daptomycin production and shortened the fermentation period both in flask and in fermentor.
authors.affiliation: Institute of Biochemistry, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, China.|Zhejiang Provincial Key Laboratory for Microbial Biochemistry and Metabolic Engineering, Hangzhou, China.
authors.forename: Peng-Hui
authors.initials: PH
authors.lastname: Yuan
authors.affiliation: Institute of Biochemistry, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, China.|Zhejiang Provincial Key Laboratory for Microbial Biochemistry and Metabolic Engineering, Hangzhou, China.
authors.forename: Ri-Cheng
authors.initials: RC
authors.lastname: Zhou
authors.affiliation: Department of Orthodontics, Hospital of Stomatology, Zhejiang University, Hangzhou, China.
authors.forename: Xuepeng
authors.initials: X
authors.lastname: Chen
authors.affiliation: Institute of Biochemistry, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, China.|Zhejiang Provincial Key Laboratory for Microbial Biochemistry and Metabolic Engineering, Hangzhou, China.
authors.forename: Shuai
authors.initials: S
authors.lastname: Luo
authors.affiliation: Institute of Biochemistry, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, China.|Zhejiang Provincial Key Laboratory for Microbial Biochemistry and Metabolic Engineering, Hangzhou, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Wang
authors.affiliation: Institute of Biochemistry, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, China xmmao@zju.edu.cn lyq@zju.edu.cn.|Zhejiang Provincial Key Laboratory for Microbial Biochemistry and Metabolic Engineering, Hangzhou, China.
authors.forename: Xu-Ming
authors.initials: XM
authors.lastname: Mao
authors.affiliation: Institute of Biochemistry, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, China xmmao@zju.edu.cn lyq@zju.edu.cn.|Zhejiang Provincial Key Laboratory for Microbial Biochemistry and Metabolic Engineering, Hangzhou, China.
authors.forename: Yong-Quan
authors.initials: YQ
authors.lastname: Li
mesh_terms: D000900:Anti-Bacterial Agents / Q000096:biosynthesis*; D018983:DNA Footprinting; D004269:DNA, Bacterial / Q000378:metabolism; D017576:Daptomycin / Q000096:biosynthesis*; D024202:Electrophoretic Mobility Shift Assay; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial; D010860:Pigments, Biological / Q000096:biosynthesis; D011485:Protein Binding; D013171:Spores, Bacterial / Q000254:growth & development; D013302:Streptomyces / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics*; D014158:Transcription, Genetic
chemical_list: D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D010860:Pigments, Biological; D014157:Transcription Factors; D017576:Daptomycin