Analysis of secretion behavior of human lysozyme from recombinant Saccharomyces cerevisiae

Mark R. Marten, Nam Soo Han, Jin Byung Park, Jin Ho Seo

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Effects of signal sequences, protein sizes and dissolved oxygen on the secretion of human lysozyme from a recombinant yeast were experimentally characterized. The systems consisted of Saccharomyces cerevisiae host SEY2102 that was transformed with two different plasmids. These plasmids were identical with an exception to the plasmid pMC614, which contained the native yeast MFα1 sequence and the plasmid pMC632 with the non-native rat α- amylase signal sequence. The expression of human lysozyme was controlled by the ADHI promoter. The native yeast MFα1 signal sequence was more efficient than the non-native rat α-amylase signal sequence in directing the secretion of human lysozyme. Lysozyme secreted with the α-amylase signal was retained inside the cells and released to the medium very slowly, thereby causing a lower cell growth rate and a decreased product secretion rate. Lysozyme was secreted more efficiently than invertase, which is an order of magnitude bigger in molecular size compared to lysozyme, which was under the direction of the MFα1 signal sequence, suggesting that protein sizes may affect the secretion efficiency. When expressed in anaerobic conditions in the medium where the ADHI promoter was derepressed, the amount of lysozyme secreted was about twice higher than that of the aerobic culture. However, the secretion rates were identical. This result showed that the dissolved oxygen level may affect the efficiency of protein secretion only, and not the secretion rate of the product protein.

Original languageEnglish
Pages (from-to)576-581
Number of pages6
JournalJournal of Microbiology and Biotechnology
Volume9
Issue number5
StatePublished - Oct 1999

Keywords

  • Human lysozyme
  • Saccharomyces cerevisiae
  • Secretion

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