Ribosome engineering and fermentation optimization leads to overproduction of tiancimycin A, a new enediyne natural product from Streptomyces sp. CB03234

  • J Ind Microbiol Biotechnol. 2018 Mar;45(3):141-151. doi: 10.1007/s10295-018-2014-8.
Ling Liu  1 ,  Jian Pan  1 ,  Zilong Wang  1 ,  Xiaohui Yan  2 ,  Dong Yang  2 ,  Xiangcheng Zhu  1  3  4 ,  Ben Shen  2  5  6 ,  Yanwen Duan  7  8  9  10 ,  Yong Huang  11  12  13
Affiliations
  • 1. Xiangya International Academy of Translational Medicine, Central South University, Changsha, 410013, Hunan, China.
  • 2. Department of Chemistry, The Scripps Research Institute, Jupiter, FL, 33458, USA.
  • 3. Xiangya Hospital, Central South University, Changsha, 410008, Hunan, China.
  • 4. Hunan Engineering Research Center of Combinatorial Biosynthesis and Natural Product Drug Discovery, Changsha, 410011, Hunan, China.
  • 5. Department of Molecular Medicine, The Scripps Research Institute, Jupiter, FL, 33458, USA.
  • 6. Natural Products Library Initiative, The Scripps Research Institute, Jupiter, FL, 33458, USA.
  • 7. Xiangya International Academy of Translational Medicine, Central South University, Changsha, 410013, Hunan, China. [email protected].
  • 8. Xiangya Hospital, Central South University, Changsha, 410008, Hunan, China. [email protected].
  • 9. Hunan Engineering Research Center of Combinatorial Biosynthesis and Natural Product Drug Discovery, Changsha, 410011, Hunan, China. [email protected].
  • 10. National Engineering Research Center of Combinatorial Biosynthesis for Drug Discovery, Changsha, 410011, Hunan, China. [email protected].
  • 11. Xiangya International Academy of Translational Medicine, Central South University, Changsha, 410013, Hunan, China. [email protected].
  • 12. Xiangya Hospital, Central South University, Changsha, 410008, Hunan, China. [email protected].
  • 13. National Engineering Research Center of Combinatorial Biosynthesis for Drug Discovery, Changsha, 410011, Hunan, China. [email protected].
PMID: 29396746 DOI: 10.1007/s10295-018-2014-8
Abstract

Tiancimycin (TNM) A, a recently discovered enediyne natural product from Streptomyces sp. CB03234, showed rapid and complete killing of Cancer cells and could be used as a payload in antibody drug conjugates. The low yield of TNM A in the wild-type strain promoted us to use ribosome engineering and fermentation optimization for its yield improvement. The Streptomyces sp. CB03234-R-16 mutant strain with a L422P mutation in RpoB, the RNA polymerase β-subunit, was obtained from the rifamycin-resistant screening. After fermentation optimization, the titers of TNM A in Streptomyces sp. CB03234-R-16 reached to 22.5 ± 3.1 mg L-1 in shaking flasks, and 13 ± 1 mg L-1 in 15 L fermentors, which were at least 40-fold higher than that in the wild-type strain (~ 0.3 mg L-1). Quantitative real-time RT-PCR revealed markedly enhanced expression of key genes encoding TNM A biosynthetic Enzymes and regulators in Streptomyces sp. CB03234-R-16. Our study should greatly facilitate the future efforts to develop TNM A into a clinical Anticancer drug.

Keywords
Enediyne natural product; In situ resin adsorption; Ribosome engineering; RpoB; Tiancimycin A.
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