Expression of an alpha-galactosidase gene under control of the homologous inulinase promoter in Kluyveromyces marxianus

Appl Microbiol Biotechnol. 1993 Nov;40(2-3):309-17. doi: 10.1007/BF00170386.

Abstract

For expression of the alpha-galactosidase gene from Cyamopsis tetragonoloba in Kluyveromyces marxianus CBS 6556 we have used the promoter of the homologous inulinase-encoding gene (INU1). The INU1 gene has been cloned and sequenced and the coding region shows an identity of 59% with the Saccharomyces cerevisiae invertase gene (SUC2). In the 5'-flanking region of INU1 we found a sequence (TAAATCCGGGG) that perfectly matches to the MIG1 binding consensus sequence (WWWWTSYGGGG) of the S. cerevisiae GAL1, GAL4 and SUC2 genes. Using the K. marxianus INU1 promoter and prepro-signal sequence, we obtained a high alpha-galactosidase production level (153 mg/l) and a secretion efficiency of 99%. Both the production level and the secretion efficiency were significantly reduced when the INU1 pro-peptide was deleted. With either the S. cerevisiae PGK or GAL7 promoter we could obtain only low alpha-galactosidase production levels (2 mg/l).

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cloning, Molecular
  • Gene Expression / physiology
  • Genes, Fungal / genetics
  • Genes, Fungal / physiology*
  • Glycoside Hydrolases / genetics*
  • Kluyveromyces / enzymology*
  • Kluyveromyces / genetics
  • Molecular Sequence Data
  • Promoter Regions, Genetic / physiology*
  • Saccharomyces cerevisiae / enzymology
  • Saccharomyces cerevisiae / genetics
  • alpha-Galactosidase / biosynthesis*
  • alpha-Galactosidase / genetics
  • alpha-Galactosidase / metabolism

Substances

  • Glycoside Hydrolases
  • alpha-Galactosidase
  • inulinase

Associated data

  • GENBANK/X68479