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

被引：22

作者：

Wu, Hang ^{[1
]}

Wang, Yansheng ^{[1
]}

Yuan, Li ^{[1
]}

Mao, Yongrong ^{[1
]}

Wang, Weiwei ^{[1
]}

Zhu, Lin ^{[1
]}

Wu, Panpan ^{[1
]}

Fu, Chengzhang ^{[2
,3
,4
]}

Mueller, Rolf ^{[3
,4
]}

Weaver, David T. ^{[1
]}

Zhang, Lixin ^{[1
,2
]}

Zhang, Buchang ^{[1
]}

机构：

[1] Anhui Univ, Inst Hlth Sci, Sch Life Sci, Hefei 230601, Anhui, Peoples R China

[2] Chinese Acad Sci, Inst Microbiol, CAS Key Lab Pathogen Microbiol & Immunol, 1 Beichen West Rd, Beijing 100101, Peoples R China

[3] Saarland Univ, Helmholtz Inst Pharmaceut Res, Helmholtz Ctr Infect Res, POB 15115, D-66041 Saarbrucken, Germany

[4] Saarland Univ, Dept Pharmaceut Biotechnol, POB 15115, D-66041 Saarbrucken, Germany

来源：

SYNTHETIC AND SYSTEMS BIOTECHNOLOGY | 2016年 / 1卷 / 01期

基金：

中国国家自然科学基金;

关键词：

Saccharopolyspora erythraea; Erythromycin; TetR family; SACE_3446; Regulatory network;

D O I：

10.1016/j.synbio.2016.01.004

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

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 S. erythraea. Herein, six TetR family transcriptional regulators (TFRs) belonging to three genomic context types were individually inactivated in S. erythraea A226, and one of them, SACE_3446, was proved to play a negative role in regulating erythromycin biosynthesis. EMSA 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 transcription. Furthermore, we explored the interaction relationships of SACE_ 3446 and previously identified TFRs (SACE_3986 and SACE_7301) associated with erythromycin production. Given demonstrated relatively independent regulation mode of SACE_3446 and SACE_3986 in erythromycin biosynthesis, we individually and concomitantly inactivated them in an industrial S. erythraea WB. Compared with WB, the WB.3446 and WB.3446.3986 mutants respectively displayed 36% and 65% yield enhancement of erythromycin A, following significantly elevated transcription of eryAI and ermE. When cultured in a 5 L fermentor, erythromycin A ofWB.3446 and WB.3446.3986 successively reached 4095 mg/L and 4670 mg/L with 23% and 41% production improvement relative to WB. The strategy reported here will be useful to improve antibiotics production in other industrial actinomycete. (C) 2016 Authors. Production and hosting by Elsevier B.V. on behalf of KeAi Communications Co., Ltd.

引用

页码：39 / 46

页数：8