DptR2, a DeoR-type auto-regulator, is required for daptomycin production in Streptomyces roseosporus.
Daptomycin, a novel cyclic lipopeptide antibiotic against Gram-positive bacteria, is produced by Streptomyces roseosporus. Though its biosynthetic mechanism, structural shuffling and fermentation optimization have been extensively studied, little is understood about its production regulation at the transcriptional levels. Here we reported that dptR2, encoding a DeoR-type regulator located close to the daptomycin biosynthesis gene cluster in S. roseosporus SW0702, is required for daptomycin production, but not for the expression of daptomycin gene cluster, suggesting that DptR2 was not a pathway-specific regulator. Furthermore, EMSA and qRT-PCR analysis suggested that DptR2 was positively auto-regulated by binding to its own promoter. Meanwhile, the binding sites on the dptR2 promoter were determined by a DNase I footprinting assay, and the essentiality of the inverted complementary sequences in the protected region for DptR2 binding was assessed. Our results for the first time reported the regulation of daptomycin production at the transcriptional level in S. roseosporus.
Zhejiang University, College of Life Sciences, Hangzhou 310058, China; Key Laboratory of Microbial Biochemistry and Metabolism Engineering of Zhejiang Province, Hangzhou 310058, China.
Feng
F
Wang
Zhejiang University, College of Life Sciences, Hangzhou 310058, China; Key Laboratory of Microbial Biochemistry and Metabolism Engineering of Zhejiang Province, Hangzhou 310058, China.
Ni-Ni
NN
Ren
Zhejiang University, College of Life Sciences, Hangzhou 310058, China; Key Laboratory of Microbial Biochemistry and Metabolism Engineering of Zhejiang Province, Hangzhou 310058, China.
Shuai
S
Luo
Hangzhou Huadong Medicine Group Biotechnology Institute Co., Ltd., Hangzhou 310011, China; Key Laboratory of Microbial Biochemistry and Metabolism Engineering of Zhejiang Province, Hangzhou 310058, China.
Xiao-Xia
XX
Chen
Zhejiang University, College of Life Sciences, Hangzhou 310058, China; Key Laboratory of Microbial Biochemistry and Metabolism Engineering of Zhejiang Province, Hangzhou 310058, China. Electronic address: xmmao@zju.edu.cn.
Xu-Ming
XM
Mao
Zhejiang University, College of Life Sciences, Hangzhou 310058, China; Key Laboratory of Microbial Biochemistry and Metabolism Engineering of Zhejiang Province, Hangzhou 310058, China. Electronic address: lyq@zju.edu.cn.
Yong-Quan
YQ
Li
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D017576:Daptomycin / Q000096:biosynthesis*; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011485:Protein Binding / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D012097:Repressor Proteins; D017576:Daptomycin
Auto-regulation; Daptomycin production; DptR2; Streptomyces; Transcriptional regulator