Strain construction for enhanced production of spinosad via intergeneric protoplast fusion

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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.

Le spinosade fait partie d’une nouvelle classe d’insecticides produits par Saccharopolyspora spinosa. Le but de cette étude était de construire une souche utilisatrice l’amidon qui produit de grandes quantités de spinosade en réalisant une fusion intergénétique entre S. spinosa et Streptomyces avermitilis. La fusion de protoplastes est une technique importante dans l’ingénierie des souches microbiennes, notamment pour les microorganismes pour lesquels il n’existe que peu d’outils moléculaires disponibles. La fusion de protoplastes a été réalisée avec les protoplastes de S. spinosa et S. avermitilis irradiés aux UV. Parmi les 76 recombinants criblés par ESI-MS et par HPLC, la souche utilisatrice d’amidon F17, identifiée en tant que S. spinosa, a été obtenue. Le rendement en production de spinosade chez F17 était augmenté de 447.22 % comparativement à la souche sauvage. Ceci constitue le premier rapport sur la fusion de protoplastes intergénétique entre S. spinosa et S. avermitilis, qui révèle de grandes applications industrielles potentielles.

Document Type: Research Article

Publication date: September 1, 2009

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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