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  • Title: Expression of an alpha-galactosidase gene under control of the homologous inulinase promoter in Kluyveromyces marxianus.
    Author: Bergkamp RJ, Bootsman TC, Toschka HY, Mooren AT, Kox L, Verbakel JM, Geerse RH, Planta RJ.
    Journal: Appl Microbiol Biotechnol; 1993 Nov; 40(2-3):309-17. PubMed ID: 7764385.
    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).
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