Purification and properties of two type-B alpha-L-arabinofuranosidases produced by Penicillium chrysogenum.
Two distinct extracellular alpha-L-arabinofuranosidases (AFases; EC 3.2.1.55) were purified from the culture filtrate of Penicillium chrysogenum 31B. The molecular masses of the enzymes were estimated to be 79 kDa (AFQ1) and 52 kDa (AFS1) by SDS-PAGE. Both enzymes had their highest activities at 50 degrees C and were stable up to 50 degrees C. Enzyme activities of AFQ1 and AFS1 were highest at pH 4.0 to 6.5 and pH 3.3 to 5.0, respectively. Addition of 10 mg/ml arabinose to the reaction mixture decreased the AFS1 activity but hardly affected AFQ1. Both enzymes displayed broad substrate specificities; they released arabinose from branched arabinan, debranched arabinan, arabinoxylan, arabinogalactan, and arabino-oligosaccharides. AFS1 also showed low activity towards p-nitrophenyl-beta-D-xylopyranoside. An exo-arabinanase, which catalyzes the release of arabinobiose from linear arabinan at the nonreducing terminus, acted synergistically with both enzymes to produce L-arabinose from branched arabinan.
Sakamoto
T
T
Division of Applied Biochemistry, Graduate School of Agriculture and Biological Sciences, Osaka Prefecture University, Osaka 599-8531, Japan. sakamoto@biochem.osakafu-u.ac.jp
Kawasaki
H
H
D000595:Amino Acid Sequence; D005656:Fungal Proteins / Q000302:isolation & purification*; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D007527:Isoenzymes / Q000302:isolation & purification; D008969:Molecular Sequence Data; D010408:Penicillium chrysogenum / Q000201:enzymology*; D013379:Substrate Specificity
D005656:Fungal Proteins; D007527:Isoenzymes; D006026:Glycoside Hydrolases; C019713:alpha-N-arabinofuranosidase