Article

Production of Aspergillus niger β-mannosidase in Pichia pastoris.

Laboratory of Biotransformation, Institute of Microbiology, Academy of Sciences of the Czech Republic, Vídeňská 1083, 142 20 Prague, Czech Republic; Department of Biochemistry, Faculty of Science, Charles University in Prague, Hlavova 2030, 128 40 Prague, Czech Republic.
Protein Expression and Purification (impact factor: 1.59). 08/2012; 85(2):159-64. DOI:10.1016/j.pep.2012.07.012 pp.159-64
Source: PubMed

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 0.44mmol/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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Keywords

933 amino acid residues
 
Aspergillus niger
 
cDNA corresponding
 
exoglycosidase specific
 
open reading frame
 
p-nitrophenyl β-d-mannopyranoside
 
recombinant proteins
 
terminal β-linked mannoside
 
yeast Pichia pastoris
 
β-Mannosidase
 
β-mannosidase gene
 

Barbora Fliedrová