Production of Aspergillus niger β-mannosidase in Pichia pastoris.

Barbora Fliedrová, Daniela Gerstorferová, Vladimír Křen, Lenka Weignerová

Laboratory of Biotransformation, Institute of Microbiology, Academy of Sciences of the Czech Republic, Vídeňská 1083, Prague, Czech Republic.

Protein expression and purification 2012 Oct

filter terms:

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). Copyright © 2012 Elsevier Inc. All rights reserved.

Citation

Barbora Fliedrová, Daniela Gerstorferová, Vladimír Křen, Lenka Weignerová. Production of Aspergillus niger β-mannosidase in Pichia pastoris. Protein expression and purification. 2012 Oct;85(2):159-64