Facile production of Aspergillus niger α-N-acetylgalactosaminidase in yeast

0377419 - MBÚ 2013 RIV US eng J - Journal Article
Mrázek, Hynek - Benada, Oldřich - Man, Petr - Vaněk, O. - Křen, Vladimír - Bezouška, Karel - Weignerová, Lenka
Facile production of Aspergillus niger α-N-acetylgalactosaminidase in yeast.
Protein Expression and Purification. Roč. 81, č. 1 (2012), s. 106-114. ISSN 1046-5928. E-ISSN 1096-0279
R&D Projects: GA MŠMT 1M0505; GA ČR GAP207/10/0321; GA ČR GD305/09/H008; GA ČR GA303/09/0477
Institutional research plan: CEZ:AV0Z50200510
Keywords : Aspergillus niger * alpha-N-Acetylgalactosaminidase * Expression
Subject RIV: CE - Biochemistry
Impact factor: 1.429, year: 2012

a-N-Acetylgalactosaminidase (a-GalNAc-ase; EC.3.2.1.49) is an exoglycosidase specific for the hydrolysis of terminal α-linked N-acetylgalactosamine in various sugar chains. The cDNA corresponding to the α-GalNAc-ase gene was cloned from Aspergillus niger, sequenced, and expressed in the yeast Saccharomyces cerevisiae. The α-GalNAc-ase gene contains an open reading frame which encodes a protein of 487 amino acid residues. The molecular mass of the mature protein deduced from the amino acid sequence of this reading frame is 54 kDa. The recombinant protein was purified to apparent homogeneity and biochemically characterized (pI 4.4, KM 0.56 mmol/l for 2-nitrophenyl 2-acetamido-2-deoxy-α-D-galactopyranoside, and optimum enzyme activity was achieved at pH 2.0–2.4 and 50–55oC). Its molecular weight was determined by analytical ultracentrifuge measurement and dynamic light scattering
Permanent Link: http://hdl.handle.net/11104/0209578