| Title: | "Molecular cloning and enzymatic characterization of a Trichoderma reesei 1,2-alpha-D-mannosidase" |
| Author(s): | Maras M; Callewaert N; Piens K; Claeyssens M; Martinet W; Dewaele S; Contreras H; Dewerte I; Penttila M; Contreras R; |
| Address: | "Department of Molecular Biology, Flanders Interuniversity Institute for Biotechnology, Ghent, Belgium" |
| DOI: | 10.1016/s0168-1656(99)00222-9 |
| ISSN/ISBN: | 0168-1656 (Print) 0168-1656 (Linking) |
| Abstract: | "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" |
| Keywords: | "Amino Acid Sequence Aspergillus/enzymology *Cloning, Molecular DNA, Complementary Mannosidases/chemistry/*genetics/*metabolism Mating Factor Molecular Sequence Data Penicillium/enzymology Peptides/genetics Pichia/enzymology/genetics Protein Sorting Signal;" |
| Notes: | "MedlineMaras, M Callewaert, N Piens, K Claeyssens, M Martinet, W Dewaele, S Contreras, H Dewerte, I Penttila, M Contreras, R eng Research Support, Non-U.S. Gov't Netherlands 2000/02/22 J Biotechnol. 2000 Feb 17; 77(2-3):255-63. doi: 10.1016/s0168-1656(99)00222-9" |
