- Production of Aspergillus niger β-mannosidase in Pichia pastoris.
Production of Aspergillus niger β-mannosidase in Pichia pastoris.
Protein expression and purification (2012-08-14)
Barbora Fliedrová, Daniela Gerstorferová, Vladimír Křen, Lenka Weignerová
PMID22884703
摘要
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).
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Sigma-Aldrich
β-Mannosidase from Helix pomatia, 5-30 units/mL, ammonium sulfate suspension, crude extract