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ACS Synth Biol. 2018 Feb 16;7(2):522-530. doi: 10.1021/acssynbio.7b00318. Epub 2017 Nov 9.

An Autoregulated Fine-Tuning Strategy for Titer Improvement of Secondary Metabolites Using Native Promoters in Streptomyces.

Author information

1
State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences , No.1 West Beichen Road, Chaoyang District, Beijing 100101, China.
2
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences , No. 2 Yuanmingyuan West Road, Haidian District, Beijing 100193, China.
3
University of Chinese Academy of Sciences , No.19A Yuquan Road, Beijing 100049, China.

Abstract

Streptomycetes are well-known producers of biologically active secondary metabolites. Various efforts have been made to increase productions of these metabolites, while few approaches could well coordinate the biosynthesis of secondary metabolites and other physiological events of their hosts. Here we develop a universal autoregulated strategy for fine-tuning the expression of secondary metabolites biosynthetic gene clusters (BGCs) in Streptomyces species. First, inducible promoters were used to control the expression of secondary metabolites BGCs. Then, the optimal induction condition was determined by response surface model in both dimensions of time and strength. Finally, native promoters with similar transcription profile to the inducible promoter under the optimal condition were identified based on time-course transcriptome analyses, and used to replace the inducible promoter following an elaborate replacement approach. The expression of actinorhodin (Act) and heterogeneous oxytetracycline (OTC) BGCs were optimized in Streptomyces coelicolor using this strategy. Compared to modulating the expression via constitutive promoters, our strategy could dramatically improve the titers of Act and OTC by 1.3- and 9.1-fold, respectively. The autoregulated fine-tuning strategy developed here opens a novel route for titer improvement of desired secondary metabolites in Streptomyces.

KEYWORDS:

Streptomyces; autoregulation; biosynthetic gene cluster; fine-tuning; inducible promoter; native promoter

PMID:
29087698
DOI:
10.1021/acssynbio.7b00318
[Indexed for MEDLINE]

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