S. Kaneko et al., PURIFICATION AND SOME PROPERTIES OF INTRACELLULAR ALPHA-L-ARABINOFURANOSIDASE FROM ASPERGILLUS-NIGER 5-16, Bioscience, biotechnology, and biochemistry, 57(7), 1993, pp. 1161-1165
Alpha-L-arabinofuranosidase was purified from a cell-free extract of A
spergillus niger 5-16 by chromatographies on DEAE-Toyopearl, SP-Toyope
arl, Ultro-gel AcA 44, Mono P, and TSK-Gel G3000SW. The final preparat
ion thus obtained showed a single band on SDS-polyacrylamide gel elect
rophoresis. The molecular weight and isoelectric point were 67,000 by
SDS-polyacrylamide gel electrophoresis and pH 3.5 by isoelectric focus
ing. The alpha-L-arabinofuranosidase contained amino acids in the orde
r of Asx > Gly > Ala > Thr > Glx = Ser. The enzyme had maximum activit
y at pH 4.0 and 60-degrees-C, and was stable from pH 4 to 7 and at tem
peratures up to 30-degrees-C. The enzyme activity was not affected con
siderably by either metal ions or chemical reagents. The enzyme releas
ed arabinose from p-nitrophenyl-alpha-L-arabinofuranoside, >3)-O-beta-
D-xylopyranosyl-(1-->4)-D-xylopyranose, and arabinan, but not from L-a
rabinofuranosyl-(1-->3)!-O-beta-D-xylopyranosyl (1-->4)-D-xylopyranose
, -L-arabinofuranosyl-(1-->3)-O-beta-D-xylopyranosyl >4)-O-beta-D-xylo
pyranosyl-(1-->4)-D-xylopyranose, gum arabic, or arabinoxylan. The lim
it of hydrolysis of arabinan was about 58% even when the enzyme was su
fficiently in excess.