Wei sheng wu xue bao = Acta microbiologica Sinica · 2013
[Rational screen of high kelimycin-producing strain by combined conventional mutagenesis and high-throughput screen method].
Оценочное исследованиеСтатьяПоддержка исследования: вне правительства СШАНазвание и аннотация: Перевод пока не подготовлен
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Liu Xinxing, Li Ping, Zhao Xiaofeng, Wang Yonghong, Zhang Siliang
Аннотация
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.
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.