An alpha-L-arabinofuranosidase from Penicillium purpurogenum: production, purification and properties.
Penicillium purpurogenum secretes arabinofuranosidase to the growth medium. Highest levels of enzyme (1.0 U ml(-1)) are obtained when L-arabitol is used as carbon source, while 0.85 and 0.7 U ml(-1) are produced with sugar beet pulp and oat spelts xylan, respectively. By means of a zymogram, three bands with arabinofuranosidase activity have been detected in the supernatant of a culture grown in oat spelts xylan. One of the enzymes was purified to homogeneity from this supernatant using gel filtration (BioGel P-100), cation exchange chromatography (CM-Sephadex C-50), hydrophobic interaction chromatography (phenyl agarose) and a second BioGel P-100 column. The enzyme is a monomer of 58 kDa with a pI of 6.5. Optimum pH is 4.0 and optimal temperature 50 degrees C. The arabinofuranosidase is highly specific for alpha-L-arabinofuranosides and liberates arabinose from arabinoxylan. The enzyme shows hyperbolic kinetics towards p-nitrophenyl-alpha-L-arabinofuranoside with a K(M) of 1.23 mM. A 36-residue N-terminal sequence is over 70% identical to that of fungal arabinofuranosidases belonging to family 54 of the glycosyl hydrolases. Based on the sequence similarity and other biochemical properties it is proposed that the purified enzyme from P. purpurogenum belongs to family 54.
De Ioannes
P
P
Laboratorio de Bioquímica, Departamento de Genética Molecular y Microbiología, Pontificia Universidad Católica de Chile, Santiago.
Peirano
A
A
Steiner
J
J
Eyzaguirre
J
J
D000595:Amino Acid Sequence; D002244:Carbon / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D007525:Isoelectric Focusing; D007527:Isoenzymes / Q000302:isolation & purification; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010407:Penicillium / Q000201:enzymology*; D020539:Sequence Analysis, Protein; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
D007527:Isoenzymes; D002244:Carbon; D006026:Glycoside Hydrolases; C019713:alpha-N-arabinofuranosidase