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

Ling Liu; Jian Pan; Zilong Wang; Xiaohui Yan; Dong Yang; Xiangcheng Zhu; Ben Shen; Yanwen Duan; Yong Huang

doi:10.1007/s10295-018-2014-8

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. Epub 2018 Feb 2.

Authors

Ling Liu¹, Jian Pan¹, Zilong Wang¹, Xiaohui Yan², Dong Yang², 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. ywduan66@sina.com.
⁸ Xiangya Hospital, Central South University, Changsha, 410008, Hunan, China. ywduan66@sina.com.
⁹ Hunan Engineering Research Center of Combinatorial Biosynthesis and Natural Product Drug Discovery, Changsha, 410011, Hunan, China. ywduan66@sina.com.
¹⁰ National Engineering Research Center of Combinatorial Biosynthesis for Drug Discovery, Changsha, 410011, Hunan, China. ywduan66@sina.com.
¹¹ Xiangya International Academy of Translational Medicine, Central South University, Changsha, 410013, Hunan, China. jonghuang@csu.edu.cn.
¹² Xiangya Hospital, Central South University, Changsha, 410008, Hunan, China. jonghuang@csu.edu.cn.
¹³ National Engineering Research Center of Combinatorial Biosynthesis for Drug Discovery, Changsha, 410011, Hunan, China. jonghuang@csu.edu.cn.

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.

MeSH terms

Adsorption
Antineoplastic Agents / metabolism
Biological Products / metabolism*
Chemistry, Pharmaceutical / methods
DNA-Directed RNA Polymerases / metabolism
Drug Design
Enediynes / metabolism*
Fermentation*
Industrial Microbiology / methods
Mutation
Real-Time Polymerase Chain Reaction
Ribosomes / chemistry*
Rifamycins / chemistry*
Streptomyces / genetics*

Substances

Antineoplastic Agents
Biological Products
Enediynes
Rifamycins
tiancimycin A
rifamycin SV
DNA-Directed RNA Polymerases
RNA polymerase beta subunit

Abstract

MeSH terms

Substances

Grants and funding