Cell wall inulinase (EC 188.8.131.52) 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. © 1991.
Laloux, O., Cassart, J. P., Delcour, J., Van Beeumen, J., & Vandenhaute, J. (1991). Cloning and sequencing of the inulinase gene of Kluyveromyces marxianus var. marxianus ATCC 12424. FEBS Letters, 289(1), 64–68. https://doi.org/10.1016/0014-5793(91)80909-M