Cloning and expression of Lipomyces starkeyi dextranase-encoding gene in yeasts

  • Kang, Hee-Kyoung (Engineering Research Institute, Chonnam National University) ;
  • Park, Ji-Young (Department of Materials and Biochemical Engineering and Institute of Bioindustrial Technology, Chonnam National University) ;
  • An, Joon-Seob (Department of Molecular Biotechnology, Chonnam National University) ;
  • Kim, Seung-Heuk (Lifenza Co. Ltd) ;
  • Kim, Do-Man (School of Biological Sciences and Technology, and The Research Institute for Catalysis, Chonnam National University, Biology Research Center for Industrial Accelerator, Dongshin University)
  • Published : 2005.04.15

Abstract

Lipomyces starkeyi produces a novel glucanhydrolase containing endo-dextranase and ${\alpha}-amylase$ activities. A cDNA from L. starkeyi encoding a dextranase was isolated and characterized. The 2,052 kb cDNA fragment (lsd1) carrying dextranase gene showed one open reading frame (ORF) composed of 1,824 bp flanked by a 41 bp 5'-UTR and a 184 bp 3'-UTR including a poly(A) tail of 27 bp. The ORF encodes for a 608 amino acid with a predicted molecular mass of 67.6 kDa. There was 77% deduced amino acid sequence identity between the LSD1 dextranase and the dextranase from Penicillium minioluteum. The primary structure of the dextranase from L. starkeyi has distant similarity with enzymes belonging to glycosyl hydrolase family 49. The lsd1 protein was expressed in the Saccharmyces cerevisiae under control of GAL1 promoter and active dextranase was produced.

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