title: Cloning and sequencing of the inulinase gene of Kluyveromyces marxianus var. marxianus ATCC 12424.
abstract: Cell wall inulinase (EC 3.2.1.7) was purified 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 1) 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 beta-fructosidases with different substrate preferences.
authors.affiliation: Lab. Génétique Moléculaire, Facultés Universitaires Notre-Dame de la Paix, Namur, Belgium.
authors.forename: O
authors.initials: O
authors.lastname: Laloux
authors.forename: J P
authors.initials: JP
authors.lastname: Cassart
authors.forename: J
authors.initials: J
authors.lastname: Delcour
authors.forename: J
authors.initials: J
authors.lastname: Van Beeumen
authors.forename: J
authors.initials: J
authors.lastname: Vandenhaute
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002473:Cell Wall / Q000201:enzymology; D003001:Cloning, Molecular; D004271:DNA, Fungal; D006026:Glycoside Hydrolases / Q000235:genetics* / Q000302:isolation & purification; D007716:Kluyveromyces / Q000201:enzymology*; D008969:Molecular Sequence Data; D012689:Sequence Homology, Nucleic Acid
chemical_list: D004271:DNA, Fungal; D006026:Glycoside Hydrolases; C023086:inulinase