Purification and some properties of intracellular alpha-L-arabinofuranosidase from Aspergillus niger 5-16.
alpha-L-Arabinofuranosidase was purified from a cell-free extract of Aspergillus niger 5-16 by chromatographies on DEAE-Toyopearl, SP-Toyopearl, Ultro-gel AcA 44, Mono P, and TSK-Gel G3000SW. The final preparation thus obtained showed a single band on SDS-polyacrylamide gel electrophoresis. The molecular weight and isoelectric point were 67,000 by SDS-polyacrylamide gel electrophoresis and pH 3.5 by isoelectric focusing. The alpha-L-arabinofuranosidase contained amino acids in the order of Asx > Gly > Ala > Thr > Glx = Ser. The enzyme had maximum activity at pH 4.0 and 60 degrees C, and was stable from pH 4 to 7 and at temperatures up to 30 degrees C. The enzyme activity was not affected considerably by either metal ions or chemical reagents. The enzyme released arabinose from p-nitrophenyl-alpha-L-arabinofuranoside, O-alpha-L-arabinofuranosyl-(1-->3)-O-beta-D-xylopyranosyl-(1-->4)-D- xylopyranose, and arabinan, but not from O-beta-D-xylopyranosyl-(1-->4)-O-[alpha-L-arabinofuranosyl- (1-->3)]-O-beta-D-xylopyranosyl-(1-->4)-D-xylopyranose, O-beta-D-xylopyranosyl-(1-->2)-O-alpha-L-arabinofuranosyl-(1-->3)-O-beta -D- xylopyranosyl-(1-->4)-O-beta-D-xylopyranosyl-(1-->4)-D-xylopyranose, gum arabic, or arabinoxylan. The limit of hydrolysis of arabinan was about 58% even when the enzyme was sufficiently in excess.
Kaneko
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S
Institute of Applied Biochemistry, University of Tsukuba, Ibaraki, Japan.
Shimasaki
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T
Kusakabe
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I
D000596:Amino Acids / Q000032:analysis; D001089:Arabinose / Q000378:metabolism; D001234:Aspergillus niger / Q000201:enzymology*; D002240:Carbohydrate Sequence; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007526:Isoelectric Point; D008670:Metals / Q000494:pharmacology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009844:Oligosaccharides / Q000378:metabolism*; D011134:Polysaccharides / Q000378:metabolism*; D013379:Substrate Specificity; D013696:Temperature
D000596:Amino Acids; D008670:Metals; D009844:Oligosaccharides; D011134:Polysaccharides; D001089:Arabinose; D006026:Glycoside Hydrolases; C019713:alpha-N-arabinofuranosidase