Biotechnology and Bioprocess Engineering:BBE

  • 제9권6호
  • /
  • Pages.523-527
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  • 2004
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  • 1226-8372(pISSN)
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  • 1976-3816(eISSN)
한국생물공학회 (The Korean Society for Biotechnology and Bioengineering)
권호 표지

Selection of Optimum Expression System for Production of Kringle Fragment of Human Apolipoprotein(a) in Saccharomyces cerevisiae

  • Cha Kwang Hyun (Department of Agricultural Biotechnology and Center for Agricultural Biotechnology, Seoul National University) ;
  • Kim Myoung Dong (Department of Agricultural Biotechnology and Center for Agricultural Biotechnology, Seoul National University) ;
  • Lee Tae Hee (Department of Agricultural Biotechnology and Center for Agricultural Biotechnology, Seoul National University) ;
  • Lim Hyung Kweon (Mogam Biotechnology Research Institute) ;
  • Jung Kyung Hwan (Department of Food and Biotechnology, Chungju National University) ;
  • Seo Jin Ho (Department of Agricultural Biotechnology and Center for Agricultural Biotechnology, Seoul National University)
  • 발행 : 2004.11.01
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초록

Recombinant Saccharomyces cerevisiae expression systems were developed to pro­duce a novel human anti-angiogenic protein called LK8, an 86 amino-acid kringle fragment pro­tein with three disulfide linkages. Galactose-inducible LK8 expression plasmid was constructed, and LK8 production levels by four S. cerevisiae strains were compared in order to select an op­timal host strain. S. cerevisiae 2805 was the most efficient among the strains tested. Elevating the LK8 gene copy number through multiple integration using 8-sequences as target sites re­sulted in more than a two-fold increase in the LK8 production level compared with the plasmid­based expression system. The maximum LK8 protein concentration of 25 mg/L was obtained from batch cultivation of the yeast transformant that harbors 16 copies of the LK8 gene. In con­clusion, the strain integrated with the multiple LK8 gene secreted the protein with relatively high yield, although, the increased LK8 gene dosage over 11 copies did not lead to further en­hancement in batch cultivations.

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