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Inactivation of SACE_3446, a tetr family transcriptional regulator, stimulates erythromycin production in Saccharopolyspora erythraea

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Synthetic and Systems Biotechnology 2016年第1期1卷 39-46页

作者： Hang Wu Yansheng Wang Li Yuan Yongrong Mao Weiwei Wang Lin Zhu Panpan Wu Chengzhang Fu Rolf Muller David T.Weaver Lixin Zhang Buchang Zhang Institute of Health Sciences School of Life SciencesAnhui UniversityHefei 230601China CAS Key Laboratory of Pathogenic Microbiology&Immunology Institute of MicrobiologyChinese Academy of SciencesBeijing 100101China Helmholtz Institute for Pharmaceutical Research Helmholtz Centre for Infection Research and Department of Pharmaceutical BiotechnologySaarland UniversityP.O.Box 1511566041 SaarbruckenGermany

Erythromycin A is a widely used antibiotic produced by Saccharopolyspora erythraea;however,its biosynthetic cluster lacks a regulatory gene,limiting the yield enhancement via regulation engineering of ***,six tetr family transcriptional regulators(TFRs)belonging to three genomic context types were individually inactivated in *** A226,and one of them,SACE_3446,was proved to play a negative role in regulating erythromycin *** and qRT-PCR analysis revealed that SACE_3446 covering intact N-terminal DNA binding domain specifically bound to the promoter regions of erythromycin biosynthetic gene eryAI,the resistant gene ermE and the adjacent gene SACE_3447(encoding a longchain fatty-acid CoA ligase),and repressed their ***,we explored the interaction relationships of SACE_3446 and previously identified TFRs(SACE_3986 and SACE_7301)associated with erythromycin *** demonstrated relatively independent regulation mode of SACE_3446 and SACE_3986 in erythromycin biosynthesis,we individually and concomitantly inactivated them in an industrial *** *** with WB,the WBΔ3446 and WBΔ3446Δ3986 mutants respectively displayed 36%and 65%yield enhancement of erythromycin A,following significantly elevated transcription of eryAI and *** cultured in a 5 L fermentor,erythromycin A ofWBΔ3446 and WBΔ3446Δ3986 successively reached 4095 mg/L and 4670 mg/L with 23%and 41%production improvement relative to *** strategy reported here will be useful to improve antibiotics production in other industrial actinomycete.

关键词： Saccharopolyspora erythraea Erythromycin tetr family SACE_3446 Regulatory network

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