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

  • Applied Genetics and Regulation
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Abstract

For expression of the α-galactosidase gene from Cyamopsistetragonoloba 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 identify 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 α-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 α-galactosidase production levels (2 mg/l).

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Authors and Affiliations

  1. Laboratorium voor Biochemie en Moleculaire Biologie, Vrije Universiteit, de Boelelaan 1083, 1081 HV, Amsterdam, The Netherlands

    Ronald J. M. Bergkamp, Theo C. Bootsman, Ruud H. Geerse & Rudi J. Planta

  2. Unilever Research Laboratorium, Olivier Noortaan 120, 3133, AT Vlaardingen, The Netherlands

    Holger Y. Toschka, Arno T. A. Mooren, Linda Kox & John M. A. Verbakel

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  1. Ronald J. M. Bergkamp
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  2. Theo C. Bootsman
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  3. Holger Y. Toschka
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  4. Arno T. A. Mooren
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  5. Linda Kox
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  6. John M. A. Verbakel
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  7. Ruud H. Geerse
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  8. Rudi J. Planta
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Correspondence to: R. J. Planta

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Bergkamp, R.J.M., Bootsman, T.C., Toschka, H.Y. et al. Expression of an α-galactosidase gene under control of the homologous inulinase promoter in Kluyveromyces marxianus . Appl Microbiol Biotechnol 40, 309–317 (1993). https://doi.org/10.1007/BF00170386

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  • DOI: https://doi.org/10.1007/BF00170386

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