Molecular cloning and enzymatic characterization of a Trichoderma reesei 1,2-alfa-D-mannosidase

Marleen Maras, Nico Callewaert, Kathleen Piens, Marc Claeyssens, Wim Martinet, Sylviane Dewaele, Hans Contreras, Isabelle Dewerte, Merja Penttilä, Roland Contreras

    Research output: Contribution to journalArticleScientificpeer-review


    A cDNA encoding 1,2-alpha-D-mannosidase mds 1 from Trichoderma reesei was cloned. The largest open reading frame occupied 1571 bp. The predicted sequence contains 523 amino acid residues for a calculated molecular mass of 56,266 Da and shows high similarity to the amino acid sequences of 1,2-alpha-D-mannosidases from Aspergillus saitoi and Penicillium citrinum (51.6 and 51.0% identity, respectively). T. reesei mannosidase was produced as a recombinant enzyme in the yeast Pichia pastoris. Replacement of the N-terminal part with the prepro-signal peptide of the Saccharomyces cerevisiae alpha-mating factor resulted in high amounts of secreted enzyme. A three-step purification protocol was designed and the enzymatic properties were analyzed. The enzyme was characterized as a class-I mannosidase.
    Original languageEnglish
    Pages (from-to)255-263
    JournalJournal of Biotechnology
    Issue number2-3
    Publication statusPublished - 2000
    MoE publication typeA1 Journal article-refereed


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