title: Production of Aspergillus niger β-mannosidase in Pichia pastoris.
abstract: β-Mannosidase (EC 3.2.1.25) is an exoglycosidase specific for the hydrolysis of terminal β-linked mannoside in various sugar chains. cDNA corresponding to the β-mannosidase gene was cloned from Aspergillus niger, sequenced, and expressed in the yeast Pichia pastoris. The β-mannosidase gene contains an open reading frame which encodes the protein with 933 amino acid residues. The wild type and recombinant proteins were purified to apparent homogeneity and biochemically characterized (K(M) 0.28 and 0.44 mmol/l for p-nitrophenyl β-d-mannopyranoside, pI 4.2 and 4.0, and their pH optima were at pH 4.5 and 5.5 and 65°C, respectively).
authors.affiliation: Laboratory of Biotransformation, Institute of Microbiology, Academy of Sciences of the Czech Republic, Vídeňská 1083, Prague, Czech Republic.
authors.forename: Barbora
authors.initials: B
authors.lastname: Fliedrová
authors.forename: Daniela
authors.initials: D
authors.lastname: Gerstorferová
authors.forename: Vladimír
authors.initials: V
authors.lastname: Křen
authors.forename: Lenka
authors.initials: L
authors.lastname: Weignerová
mesh_terms: D001234:Aspergillus niger / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D005656:Fungal Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D010843:Pichia / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D013696:Temperature; D044902:beta-Mannosidase / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification
chemical_list: D003470:Culture Media; D005656:Fungal Proteins; D011994:Recombinant Proteins; D044902:beta-Mannosidase