Purification and properties of an alpha-D-xylosidase from Aspergillus niger.
Two components of alpha-D-xylosidase (alpha-D-xylosidase I and II) were detected in the culture filtrate of Aspergillus nigher grown in a medium containing Sanzyme 1000-treated Glyloid 2A. The major component (alpha-D-xylosidase I) was purified to an electrophoretically pure state. The purified enzyme showed approximately 540-fold increase in specific activity over the original culture filtrate. The purified enzyme was shown to be an oligomeric protein consisting of four subunits, each of which had a molecular weight of 123,000. The enzyme showed the highest activity at pH 2.5-3.0 and 45 degrees C, and was stable in the pH range from 3.0 to 7.0 and at the temperatures up to 60 degrees C. The isoelectric point of this enzyme was pH 5.6. The purified enzyme was highly specific for p-nitrophenyl alpha-D-xylopyranoside and isoprimeverose (6-O-alpha-D-xylopyranosyl-D-glucopyranose). The apparent Km and Vmax values of the enzyme for p-nitrophenyl alpha-D-xylopyranoside and isoprimeverose were 10.5 mM and 40.8 mumol/min/mg protein, and 2.2 mM and 30 mumol/min/mg protein, respectively. The purified enzyme could also split off the alpha-D-xylopyranosyl residue on the non-reducing terminal of the backbone of oligoxyloglucans such as alpha-D-xylopyranosyl-(1----6)-beta-D-glucopyranosyl- (1----4)-[(alpha-D-xylopyranosyl-(1----6)-]-beta-D-glucopyranosyl- (1----4)-] 2-D-glucopyranose.
D000596:Amino Acids / Q000032:analysis; D001234:Aspergillus niger / Q000201:enzymology*; D006026:Glycoside Hydrolases / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D008970:Molecular Weight; D009844:Oligosaccharides / Q000032:analysis; D013379:Substrate Specificity; D013816:Thermodynamics; D014995:Xylosidases / Q000302:isolation & purification* / Q000378:metabolism
D000596:Amino Acids; D007527:Isoenzymes; D009844:Oligosaccharides; D006026:Glycoside Hydrolases; D014995:Xylosidases; C047831:alpha-D-xylosidase