Purification and properties of a thermostable extracellular beta-D-xylosidase produced by a thermotolerant Aspergillus phoenicis

A beta -D-xylosidase was purified from cultures of a thermotolerant strain of Aspergillus phoenicis grown on xylan at 45 degreesC. The enzyme was puri fied to homogeneity by chromatography on DEAE-cellulose and Sephadex G-100. The purified enzyme was a monomer of molecular mass 132 kDa by gel filtrat ion and SDS-PAGE. Treatment with endoglycosidase H resulted in a protein wi th a molecular mass of 104 kDa, The enzyme was a glycoprotein with 43.5% ca rbohydrate content and exhibited a pi of 3.7. Optima of temperature and pH were 75 degreesC and 4.0-4.5, respectively. The activity was stable at 60 d egreesC and had a K-m of 2.36 mM for p-nitrophenyl-beta -D-xylopiranoside. The enzyme did not exhibit xylanase, cellulase, galactosidase or arabinosid ase activities. The purified enzyme was active against natural substrates, such as xylobiose and xylotriose.