Debranching of arabinoxylan: properties of the thermoactive recombinant α-L-arabinofuranosidase fromClostridium stercorarium (ArfB)
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The genearfB encoding α-L-arabinofuranosidase B of the cellulolytic thermophileClostridium stercorarium was expressed inEscherichia coli from a 2.2-kbEcoRI DNA fragment. The recombinant gene product ArfB was purified by fast-performance liquid chromatography. It has a tetrameric structure with a monomeric relative molecular mass of 52 00. The optima for temperature and pH are 70°C and 5.0 respectively. The enzyme appears to have no metal cofactor requirement and is sensitive to sulfhydryl reagents. It hydrolyzes aryl and alkyl α-L-arabinofuranosides and cleaves arabinosyl side-chains from arabinoxylan (oat-spelt xylan) and from xylooligosaccharides produced by recombinant endoxylanase XynA from the same organism. The identity of the N-terminal amino acid sequences indicates that ArfB corresponds to the major α-arabinosidase activity present in the culture supernatant ofC. stecorarium.
KeywordsXylose Arabinose Xylobiose Potassium Acetate Xylotriose
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