title: SACE_3986, a TetR family transcriptional regulator, negatively controls erythromycin biosynthesis in Saccharopolyspora erythraea.
abstract: Erythromycin, a medically important antibiotic, is produced by Saccharopolyspora erythraea. Unusually, the erythromycin biosynthetic gene cluster lacks a regulatory gene, and the regulation of its biosynthesis remains largely unknown. In this study, through gene deletion, complementation and overexpression experiments, we identified a novel TetR family transcriptional regulator SACE_3986 negatively regulating erythromycin biosynthesis in S. erythraea A226. When SACE_3986 was further inactivated in an industrial strain WB, erythromycin A yield of the mutant was increased by 54.2 % in average compared with that of its parent strain, displaying the universality of SACE_3986 as a repressor for erythromycin production in S. erythraea. qRT-PCR analysis indicated that SACE_3986 repressed the transcription of its adjacent gene SACE_3985 (which encodes a short-chain dehydrogenase/reductase), erythromycin biosynthetic gene eryAI and the resistance gene ermE. As determined by EMSA analysis, purified SACE_3986 protein specifically bound to the intergenic region between SACE_3985 and SACE_3986, whereas it did not bind to the promoter regions of eryAI and ermE. Furthermore, overexpression of SACE_3985 in A226 led to enhanced erythromycin A yield by at least 32.6 %. These findings indicate that SACE_3986 is a negative regulator of erythromycin biosynthesis, and the adjacent gene SACE_3985 is one of its target genes. The present study provides a basis to increase erythromycin production by engineering of SACE_3986 and SACE_3985 in S. erythraea.
authors.affiliation: Institute of Health Sciences, School of Life Sciences, Anhui University, Hefei, 230601, China.
authors.forename: Panpan
authors.initials: P
authors.lastname: Wu
authors.forename: Hui
authors.initials: H
authors.lastname: Pan
authors.forename: Congming
authors.initials: C
authors.lastname: Zhang
authors.forename: Hang
authors.initials: H
authors.lastname: Wu
authors.forename: Li
authors.initials: L
authors.lastname: Yuan
authors.forename: Xunduan
authors.initials: X
authors.lastname: Huang
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhou
authors.forename: Bang-ce
authors.initials: BC
authors.lastname: Ye
authors.forename: David T
authors.initials: DT
authors.lastname: Weaver
authors.forename: Lixin
authors.initials: L
authors.lastname: Zhang
authors.forename: Buchang
authors.initials: B
authors.lastname: Zhang
mesh_terms: D000900:Anti-Bacterial Agents / Q000096:biosynthesis; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D021901:DNA, Intergenic / Q000235:genetics / Q000378:metabolism; D004917:Erythromycin / Q000096:biosynthesis*; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics*; D005798:Genes, Bacterial / Q000235:genetics; D005810:Multigene Family / Q000235:genetics; D012097:Repressor Proteins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D016930:Saccharopolyspora / Q000145:classification / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D014157:Transcription Factors / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D021901:DNA, Intergenic; D012097:Repressor Proteins; D014157:Transcription Factors; D004917:Erythromycin