title: [Rational screen of high kelimycin-producing strain by combined conventional mutagenesis and high-throughput screen method].
abstract: 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%.
  
  
  CONCLUSIONS
  This study shows that the screening industrial production strains can be enhanced effectively by combining multiple conventional mutagenesis and high-throughput screen methods.
authors.affiliation: Shanghai Tonglian Pharmaceutical Technology Company Limited, Shanghai 200231, China. 446595315@qq.com
authors.forename: Xinxing
authors.initials: X
authors.lastname: Liu
authors.forename: Ping
authors.initials: P
authors.lastname: Li
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Zhao
authors.forename: Yonghong
authors.initials: Y
authors.lastname: Wang
authors.forename: Siliang
authors.initials: S
authors.lastname: Zhang
mesh_terms: D000900:Anti-Bacterial Agents / Q000096:biosynthesis*; D005285:Fermentation; D005821:Genetic Techniques; D016296:Mutagenesis; D015572:Spiramycin / Q000096:biosynthesis*; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism* / Q000528:radiation effects; D014466:Ultraviolet Rays
chemical_list: D000900:Anti-Bacterial Agents; D015572:Spiramycin