Hydrolytic properties of a β-mannosidase purified from Aspergillus niger
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Hydrolytic properties of a β-mannosidase purified from Aspergillus niger. / Ademark, Pia; Lundqvist, Jon; Hägglund, Per; Tenkanen, Maija; Torto, Nelson; Tjerneld, Folke; Stålbrand, Henrik.
I: Journal of Biotechnology, Bind 75, Nr. 2-3, 08.10.1999, s. 281-289.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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T1 - Hydrolytic properties of a β-mannosidase purified from Aspergillus niger
AU - Ademark, Pia
AU - Lundqvist, Jon
AU - Hägglund, Per
AU - Tenkanen, Maija
AU - Torto, Nelson
AU - Tjerneld, Folke
AU - Stålbrand, Henrik
PY - 1999/10/8
Y1 - 1999/10/8
N2 - A β-mannosidase was purified to homogeneity from the culture filtrate of Aspergillus niger. A specific activity of 500 nkat mg-1 and a 53-fold purification was achieved using ammonium sulfate precipitation, anion- exchange chromatography, and gel filtration. The isolated enzyme has an isoelectric point of 5.0 and appears to be a dimer composed of two 135-kDa subunits. It is a glycoprotein and contains 17% N-linked carbohydrate by weight. Maximal activity was observed at pH 2.4-5.0 and at 70°C. The β- mannosidase hydrolyzed β-1,4-linked manno-oligosaccharides of degree of polymerization (DP) 2-6 and also released mannose from polymeric ivory nut mannan and galactomannan. The K(m) and V(max) values for p-nitrophenyl-β-D- mannopyranoside were 0.30 mM and 500 nkat mg-1, respectively. Hydrolysis of D-galactose substituted manno-oligosaccharides showed that the β-mannosidase was able to cleave up to, but not beyond, a side group. An internal peptide sequence of 15 amino acids was highly similar to that of an Aspergillus aculeatus β-mannosidase belonging to family 2 of glycosyl hydrolases.
AB - A β-mannosidase was purified to homogeneity from the culture filtrate of Aspergillus niger. A specific activity of 500 nkat mg-1 and a 53-fold purification was achieved using ammonium sulfate precipitation, anion- exchange chromatography, and gel filtration. The isolated enzyme has an isoelectric point of 5.0 and appears to be a dimer composed of two 135-kDa subunits. It is a glycoprotein and contains 17% N-linked carbohydrate by weight. Maximal activity was observed at pH 2.4-5.0 and at 70°C. The β- mannosidase hydrolyzed β-1,4-linked manno-oligosaccharides of degree of polymerization (DP) 2-6 and also released mannose from polymeric ivory nut mannan and galactomannan. The K(m) and V(max) values for p-nitrophenyl-β-D- mannopyranoside were 0.30 mM and 500 nkat mg-1, respectively. Hydrolysis of D-galactose substituted manno-oligosaccharides showed that the β-mannosidase was able to cleave up to, but not beyond, a side group. An internal peptide sequence of 15 amino acids was highly similar to that of an Aspergillus aculeatus β-mannosidase belonging to family 2 of glycosyl hydrolases.
KW - β-Mannosidase
KW - Aspergillus niger
KW - Hydrolysis
KW - Manno-oligosaccharides
UR - http://www.scopus.com/inward/record.url?scp=0032845641&partnerID=8YFLogxK
U2 - 10.1016/S0168-1656(99)00172-8
DO - 10.1016/S0168-1656(99)00172-8
M3 - Journal article
C2 - 10553664
AN - SCOPUS:0032845641
VL - 75
SP - 281
EP - 289
JO - Journal of Biotechnology
JF - Journal of Biotechnology
SN - 0168-1656
IS - 2-3
ER -
ID: 240162541