Biotechnology and Bioprocess Engineering:BBE

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Flow Cytometric Analysis of Human Lysozyme Production in Recombinant Saccharomyces cerevisiae

  • Peterson Marvin S. (Signal Pharmaceuticals Inc., San Diego) ;
  • Kim Myoung-Dong (Department of Food Science and Technology and Research Center for New Bio-Materials in Agriculture, Seoul National University) ;
  • Han Ki-Cheol (Department of Food Science and Technology and Research Center for New Bio-Materials in Agriculture, Seoul National University) ;
  • Kim Ji-Hyun (Ministry of Commerce, Industry and Energy) ;
  • Seo Jin-Ho (Department of Food Science and Technology and Research Center for New Bio-Materials in Agriculture, Seoul National University)
  • Published : 2002.01.01
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Abstract

Flow cytometric techniques were used to investigate cell size, protein content and cell cycle behavior of recombinant Saccharomyces cerevisiae strains producing human lysozyme (HLZ). Two different signal sequences, the native yeast $MF\alpha1$ signal sequence and the rat $\alpha-amylase$ signal sequence, were used for secretion of HLZ. The strain containing the rat $\alpha-amylase$ signal sequence showed a higher level of internal lysozyme and lower specific growth rates. Flow cytometric analysis of the total protein content and cell size showed the strain harboring the native yeast signal sequence had a higher total protein content than the strain containing the rat $\alpha-amylase$ signal sequence. Cell cycle analysis indicated that the two lysozyme producing recombinant strains had an increased number of cells in the $G_2+M$ phase of the yeast cell cycle compared with the host strain SEY2102.

Keywords

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