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J Biotechnol


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"
Journal Title:J Biotechnol
Year:2000
Volume:77
Issue:2-Mar
Page Number:255 - 263
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"

 
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