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Applied Microbiology and Biotechnology

, Volume 103, Issue 16, pp 6645–6655 | Cite as

Heterologous production of chlortetracycline in an industrial grade Streptomyces rimosus host

  • Xuefeng Wang
  • Shouliang Yin
  • Jing Bai
  • Yang Liu
  • Keqiang Fan
  • Huizhuan Wang
  • Fang Yuan
  • Baohua ZhaoEmail author
  • Zilong LiEmail author
  • Weishan WangEmail author
Applied genetics and molecular biotechnology

Abstract

High-yielding industrial Streptomyces producer is usually obtained by multiple rounds of random mutagenesis and screening. These strains have great potential to be developed as the versatile chassis for the discovery and titer improvement of desired heterologous products. Here, the industrial strain Streptomyces rimosus 461, which is a high producer of oxytetracycline, has been engineered as a robust host for heterologous expression of chlortetracycline (CTC) biosynthetic gene cluster. First, the industrial chassis strain SR0 was constructed by deleting the whole oxytetracycline gene cluster of S. rimosus 461. Then, the biosynthetic gene cluster ctc of Streptomyces aureofaciens ATCC 10762 was integrated into the chromosome of SR0. With an additional constitutively expressed cluster-situated activator gene ctcB, the CTC titer of the engineering strain SRC1 immediately reached 1.51 g/L in shaking flask. Then, the CTC titers were upgraded to 2.15 and 3.27 g/L, respectively, in the engineering strains SRC2 and SRC3 with the enhanced ctcB expression. Further, two cluster-situated resistance genes were co-overexpressed with ctcB. The resultant strain produced CTC up to 3.80 g/L in shaking flask fermentation, which represents 38 times increase in comparison with that of the original producer. Overall, SR0 presented in this study have great potential to be used for heterologous production of tetracyclines and other type II polyketides.

Keywords

Streptomyces rimosus Tetracyclines Chlortetracycline High production Chassis 

Notes

Acknowledgments

We kindly thank Prof. Chunbo Lou (Institute of Microbiology of the Chinese Academy of Sciences) for providing the plasmid pSET153-ctc. We dedicate this article to the memory of our friend and mentor Keqian Yang, who made important contribution to improve the functionality of industrial microorganisms.

Funding information

This work was supported by the Science and Technology Program of Hebei (No. 18222916).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

253_2019_9970_MOESM1_ESM.pdf (522 kb)
ESM 1 (PDF 522 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.College of Life ScienceHebei Normal UniversityShijiazhuangPeople’s Republic of China
  2. 2.Hebei Shengxue Dacheng Pharmaceutical Co., Ltd.ShijiazhuangPeople’s Republic of China
  3. 3.School of Life SciencesNorth China University of Science and TechnologyTangshanPeople’s Republic of China
  4. 4.State Key Laboratory of Microbial ResourcesInstitute of Microbiology, Chinese Academy of SciencesBeijingPeople’s Republic of China

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