Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Cloning of a Gene Encoding Dextranase from Lipomyces starkeyi and its Expression in Pichia pastoris

  • Kang, Hee-Kyoung (School of Biological Sciences and Technology, Chonnam National University) ;
  • Park, Ji-Young (Jeonbuk Branch Institute, Korea Research Institute of Bioscience and Biotechnology) ;
  • Ahn, Joon-Seob (Gang-Jin City Agricultural Technology Extension Center) ;
  • Kim, Seung-Heuk (Lifenza Co. Ltd.) ;
  • Kim, Do-Man (School of Biological Sciences and Technology, Chonnam National University)
  • Published : 2009.02.28
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Abstract

A gene(lsd1) encoding dextranase from Lipomyces starkeyi KSM22 has been previously cloned, sequenced, and expressed in Saccharomyces cerevisiae. The gene consisting of 1,824 base pairs and encoding a protein of 608 amino acids was then cloned into and secretively expressed in Pichia pastoris under the control of the AOX1 promoter. The dextranase productivity of the P. pastoris transformant(pPIC9K-LSD1, 134,000 U/I) was approximately 4.2-fold higher than that of the S. cerevisiae transformant(pYLSD1, 32,000 U/I) cultured in an 8-1 fermentor. Over 0.63 g/l of active dextranase was secreted into the medium after methanol induction. The dextranase of the P. pastoris transformant, as analyzed by SDS-PAGE and Western blotting, showed only one homogeneous band. This dextranase of the P. pastoris transformant showed a broad band near 73 kDa. Rabbit monoclonal antibodies against a synthetic LSD1 peptide mix also recognized approximately 73 kDa.

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References

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