Adaptive evolution and addition of betaine to promote ansamitocin production

XU Wen-Long1 HU Feng-Xian1 LIU Ting1 WEI Liu-Jing1 HUA Qiang1

(1.State Key Laboratory of Bioreactor Engineering, School of Biotechnology, East China University of Science and Technology, Shanghai, China 200237)

【Abstract】[Background] Ansamitocins, macrocyclic lactam compounds related to maytansine, are synthesized by Actinosynnema pretiosum ssp. auranticum. Based on the difference of ester side chain moieties in C-3 position, ansamitocins contain a series of derivatives. Currently, a novel anticancer drug named Kadcyla (T-DM1), formed from ansamitocin P-3 (AP-3), enters the market. T-DM1 has been approved by FDA as an antibody-drug for treating breast cancer. However, the low yield of ansamitocins limits its commercial application. [Objective] To improve AP-3 yield from A. pretiosum ssp. auranticum ATCC 31565, we applied adaptive evolution and addition of betaine. [Methods] The wild-type A. pretiosum ssp. auranticum was used as original strain to the adverse stress streptomycin and paromomycin. The evolved mutants were screened with better production of AP-3. After 0.1% betaine was added into the fermentation broth of the evolved mutants, the production of AP-3 was further enhanced. Then, gene transcription levels related to AP-3 biosynthesis were analyzed in evolved mutants and original strain, to investigate the reasons for enhancement of ansamitocin. [Results] Two evolved strains Str16–4-4 and Par16–2-1 have been obtained. After seven days of fermentation, their AP-3 content was increased by 33.4% and 31.7%, respectively. Addition of 0.1% betaine ultimately resulted in the increase of 54.6% and 47.4% AP-3 production compared with the wild strain. [Conclusion] Through the adaptive evolution strategy, AP-3 production was improved in A. pretiosum ssp. auranticum. This study provides a new strategy to improve the production of AP-3, and also gives a new example for adaptive evolutionary strategy to promote the production of target products.

【Keywords】 Adaptive evolution; Actinosynnema pretiosum; Ansamitocin P-3; Betaine;

【Funds】 Research Fund for the Doctoral Program of Higher Education of China (20130074110002)

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(Translated by ZHAO B)

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This Article

ISSN:0253-2654

CN: 11-1996/Q

Vol 46, No. 07, Pages 1561-1570

July 2019

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Article Outline

Abstract

  • 1 Materials and methods
  • 2 Results and analysis
  • 3 Discussion
  • 4 Conclusions
  • References