Purification and substrate specificities of two alpha-L-arabinofuranosidases from Aspergillus awamori IFO 4033.
alpha-L-Arabinofuranosidases I and II were purified from the culture filtrate of Aspergillus awamori IFO 4033 and had molecular weights of 81,000 and 62,000 and pIs of 3.3 and 3.6, respectively. Both enzymes had an optimum pH of 4.0 and an optimum temperature of 60 degreesC and exhibited stability at pH values from 3 to 7 and at temperatures up to 60 degrees C. The enzymes released arabinose from p-nitrophenyl-alpha-L-arabinofuranoside, O-alpha-L-arabinofuranosyl-(1-->3)-O-beta-D-xylopyranosyl-(1-->4)-D-x ylopyranose, and arabinose-containing polysaccharides but not from O-beta-D-xylopyranosyl-(1-->2)-O-alpha-L-arabinofuranosyl-(1-->3)-O-b eta-D-xylopyranosyl-(1-->4)-O-beta-D-xylopyranosyl-(1-->4)-D-xylopyra nose. alpha-L-Arabinofuranosidase I also released arabinose from O-beta-D-xylopy-ranosyl-(1-->4)-[O-alpha-L-arabinofuranosyl- (1-->3)]- O-beta-D-xylopyranosyl-(1-->4)-D-xylopyranose. However, alpha-L-arabinofuranosidase II did not readily catalyze this hydrolysis reaction. alpha-L-Arabinofuranosidase I hydrolyzed all linkages that can occur between two alpha-L-arabinofuranosyl residues in the following order: (1-->5) linkage > (1-->3) linkage > (1-->2) linkage. alpha-L-Arabinofuranosidase II hydrolyzed the linkages in the following order: (1-->5) linkage > (1-->2) linkage > (1-->3) linkage. alpha-L-Arabinofuranosidase I preferentially hydrolyzed the (1-->5) linkage of branched arabinotrisaccharide. On the other hand, alpha-L-arabinofuranosidase II preferentially hydrolyzed the (1-->3) linkage in the same substrate. alpha-L-Arabinofuranosidase I released arabinose from the nonreducing terminus of arabinan, whereas alpha-L-arabinofuranosidase II preferentially hydrolyzed the arabinosyl side chain linkage of arabinan.
Kaneko
S
S
Institute of Applied Biochemistry, University of Tsukuba, 1-1-1 Tennoodai, Tsukuba, Ibaraki 305, Japan.
Arimoto
M
M
Ohba
M
M
Kobayashi
H
H
Ishii
T
T
Kusakabe
I
I
D000595:Amino Acid Sequence; D001230:Aspergillus / Q000201:enzymology*; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D013816:Thermodynamics
D007527:Isoenzymes; D009844:Oligosaccharides; D006026:Glycoside Hydrolases; C019713:alpha-N-arabinofuranosidase