Cloning and sequencing of the inulinase gene of Kluyveromyces marxianus var. marxianus ATCC 12424

FEBS Letters

Laloux, Olivier²  Cassart, Jean-Pol²  Van Beeumen, Jozef¹  Delcour, Jean²  Vandenhaute, Jean² 

[1] Lab. Microbiologie, Rijksuniversiteit, Ledeganckstraat 35, B-9000 Gent, Belgium;Lab. Génétique Moléculaire, Facultés Universitaires Notre-Dame de la Paix, rue de Bruxelles 61, 8-5000 Namur, Belgium

关键词: Inulinase;    Invertase;    Kluyveromyces marxianus var. marxianus;    Kluyveromyces fragilis;   

DOI  :  10.1016/0014-5793(91)80909-M

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Cell wall inulinase (EC 3.2.1.7) was pirified from Kluyveromyces marxianus var. marxianus (formerly K. fragilis) and its N-terminal 33-amino acid sequence was established. PCR amplification of cDNA with 2 sets of degenerate primers yielded a genomic probe which was then used to screen a genomic library established in the YEp351 yeast shuttle vector. One of the selected recombinant plasmids allowed an invertase-negative Saccharomyces cerevisiae mutant to grow on inulin. It was shown to contain an inulinase gene (INU I) encoding a 555-amino acid precursor protein with a typical N-terminal signal peptide. The sequence of inulinase displays a high similarity (67%) to S. cerevisiae invertase, suggesting a common evolutionary origin for yeast ß-fructosidases with different substrate preferences.

【 授权许可】

Unknown