title: Strain construction for enhanced production of spinosad via intergeneric protoplast fusion.
abstract: Spinosad is a new class of insecticides produced by Saccharopolyspora spinosa. The aim of this study was to construct a starch-utilizing strain that overproduced spinosad by intergeneric fusion between S. spinosa and Streptomyces avermitilis. Protoplast fusion is an important technique for engineering microbial strains, especially for microorganisms with few available molecular genetic tools. Protoplast fusion was conducted with UV-irradiated protoplasts of S. spinosa and S. avermitilis. Among 76 recombinants screened by ESI-MS and HPLC, a starch-utilizing strain F17, identified as S. spinosa, was obtained. The yield of spinosad in F17 was increased by 447.22%, compared with the yield of the wild-type strain. This is the first report of intergeneric protoplast fusion between S. spinosa and S. avermilitis, which shows great potential for industrial applications.
authors.affiliation: Department of Microbiology, China Agricultural University, Beijing, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Wang
authors.forename: XiaoLin
authors.initials: X
authors.lastname: Zhang
authors.forename: Zhi
authors.initials: Z
authors.lastname: Chen
authors.forename: Ying
authors.initials: Y
authors.lastname: Wen
authors.forename: Yuan
authors.initials: Y
authors.lastname: Song
mesh_terms: D002459:Cell Fusion; D004338:Drug Combinations; D005818:Genetic Engineering; D007306:Insecticides / Q000378:metabolism*; D018942:Macrolides / Q000378:metabolism*; D011523:Protoplasts / Q000378:metabolism*; D011995:Recombination, Genetic; D016930:Saccharopolyspora / Q000235:genetics* / Q000378:metabolism / Q000528:radiation effects; D013213:Starch / Q000378:metabolism; D013302:Streptomyces / Q000235:genetics* / Q000528:radiation effects; D014466:Ultraviolet Rays
chemical_list: D004338:Drug Combinations; D007306:Insecticides; D018942:Macrolides; D013213:Starch; C415329:spinosad