Biotechnology and Bioprocess Engineering:BBE

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Effect of a PMR1 Disruption on the Processing of Heterologous Glycoproteins Secreted in the Yeast Saccharomyces cerevisiae

  • Kim, Moo-Woong (Korea Research Institute of Bioscience and Biotechnology, PO Box 115, Yusong, Taejon 305-600) ;
  • Ko, Su-Min (Korea Research Institute of Bioscience and Biotechnology, PO Box 115, Yusong, Taejon 305-600) ;
  • Kim, Jeong-Yoon (Department of Microbiology, Chungnam National University, Taejon 305-764) ;
  • Sohn, Jung-Hoon (Korea Research Institute of Bioscience and Biotechnology, PO Box 115, Yusong, Taejon 305-600) ;
  • Park, Eui-Sung (Korea Research Institute of Bioscience and Biotechnology, PO Box 115, Yusong, Taejon 305-600) ;
  • Kang, Hyun-Ah (Korea Research Institute of Bioscience and Biotechnology, PO Box 115, Yusong, Taejon 305-600) ;
  • Rhee, Sang-Ki (Korea Research Institute of Bioscience and Biotechnology, PO Box 115, Yusong, Taejon 305-600)
  • Published : 2000.07.01
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Abstract

The Saccharomyces cerevisiae PMR1 gene encodes a Ca2+-ATPase localized in the Golgi. We have investigated the effects of PMR1 disruption in S. cerevisiae on the glycosylation and secretion of three heterologous glycoproteins, human ${\alpha}$1-antitrypsin (${\alpha}$1-AT), human antithrombin III (ATHIII), and Aspergillus niger glucose oxidase (GOD). The pmr1 null mutant strain secreted larger amounts of ATHIII and GOD proteins per a unit cell mass than the wild type strain. Despite a lower growth rate of the pmr1 mutant, two-fold higher level of human ATHIII was detected in the culture supernatant from the pmr1 mutant compared to that of the wild-type strain. The pmr1 mutant strain secreted ${\alpha}$1-AT and the GOD proteins mostly as core-glycosylated forms, in contrast to the hyperglycosylated proteins secreted in the wild-type strain. Furthermore, the core-glycosylated forms secreted in the pmr1 mutant migrated slightly faster on SDS-PAGE than those secreted in the mnn9 deletion mutant and the wild type strains. Analysis of the recombinant GOD with anti-${\alpha}$1,3-mannose antibody revealed that GOD secreted in the pmr1 mutant did not have terminal ${\alpha}$1,3-linked mannose unlike those secreted in the mnn9 mutant and the wild type strains. The present results indicate that the pmr1 mutant, with the super-secretion phenotype, is useful as a host system to produce recombinant glycoproteins lacking high-mannose outer chains.

Keywords

References

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