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Updated: May 6, 2026
A Method for Screening and Validation of Resistant Mutations Against Kinase Inhibitors
Published on: December 7, 2014
[Rational screen of high kelimycin-producing strain by combined conventional mutagenesis and high-throughput screen
Xinxing Liu1, Ping Li, Xiaofeng Zhao
1Shanghai Tonglian Pharmaceutical Technology Company Limited, Shanghai 200231, China. 446595315@qq.com
Unlabelled:
Kelimycin, is a new macrolide antibiotic drug obtained through genetic engineering approaches. With 4"-O-isovalerylspiramycins as the major components, was produced by genetically engineered Streptomyces spiramyceticus transformed with 4"-O-acyltransferase gene from S. mycarofaciens.
Objective:
Improve the efficiency of strain fermentation, to meet the needs of industrial production.
Methods:
The enhanced kelimycin-producing strain was obtained by applying various conventional mutagenesis approaches, and high-throughput screen methods, including protoplast mutagenesis by ultraviolet, mutagenesis by diethyl sulfate and UV-reactivation, valine content resistance screen and enrichment of improved strains. A strategy for positive mutant enrichment was developed after mutagenesis and before high-throughput screen.
Results:
Finally, the high-producing strain WSJ-1-7-49-133-82-18-43 was obtained and its potency in shake flask increased by 56% compared to the original strain. The potency in 500 L pilot fermenter increased by 61%.
Conclusion:
This study shows that the screening industrial production strains can be enhanced effectively by combining multiple conventional mutagenesis and high-throughput screen methods.
