Purification and characterization of two acetyl xylan esterases from Penicillium purpurogenum.
Penicillium purpurogenum produces several enzymes active in xylan hydrolysis, of there, the acetyl xylan esterase (AXE) activity secreted by the fungus has now been studied. The amount of activity obtained in the culture is related to the degree of acetylation of the carbon source used, the best being chemically acetylated xylan. AXE was concentrated from culture supernatants by ultrafiltration and (NH4)2SO4 precipitation and fractionated by gel filtration in Bio-Gel P-300. Two peaks of activity (AXE I and AXE II) were obtained. These two enzymes were further purified separately to homogeneity by chromatography in CM-Sephadex C-50 and chromatofocusing. AXE I (M(r) 48,000) has a pl of 7.5, while AXE II (M(r) 23,000) has a pl of 7.8. Optimal enzyme activity was at pH 5.3 and 50 degrees C for AXE I and pH 6.0 and 60 degrees C for AXE II. Both enzymes are active towards several acetylated substrates. Antisera against the two enzymes do not cross-react, and the N-terminal sequences of AXE I and II do not show similarities. These results suggest that AXE I and AXE II are the products of different genes.
Egaña
L
L
Laboratorio de Bioquímica, Pontificia Universidad Católica de Chile, Santiago, Chile.
Gutiérrez
R
R
Caputo
V
V
Peirano
A
A
Steiner
J
J
Eyzaguirre
J
J
D000115:Acetylesterase / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D000595:Amino Acid Sequence; D001709:Biotechnology; D005800:Genes, Fungal; D006863:Hydrogen-Ion Concentration; D007120:Immunochemistry; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010407:Penicillium / Q000201:enzymology* / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D013696:Temperature
D000115:Acetylesterase; C081851:acetylxylan esterase