title: Purification and characterization of extracellular phytase from Aspergillus niger ATCC 9142.
abstract: Extracellular phytase produced by Aspergillus niger ATCC 9142 was purified to homogeneity by employing an initial ultrafiltration step, followed by chromatography using ion exchange, gel filtration and chromatofocusing steps. The purified enzyme was an 84 kDa, monomeric protein. It possessed a temperature optimum of 65 degrees C, and a pH optimum of 5.0. Km and Vmax values of 100 microM and 7 nmol/s, respectively, were recorded and these values fall well within the range of those previously reported for microbial phytases. Substrate specificity studies indicated that, while the enzyme could hydrolyse a range of non-phytate-based phosphorylated substrates, its preferred substrate was phytate. Phytase activity was moderately stimulated in the presence of Mg2+, Mn2+, Cu2+, Cd2+, Hg2+, Zn2+ and F- ions. Activity was not significantly affected by Fe2- or Fe3- and was moderately inhibited by Ca2+. The enzyme displayed higher thermostability at 80 degrees C than did two commercial phytase products. Initial characterisation of the purified enzyme suggested that it could be a potential candidate for use as an animal feed supplement.
authors.affiliation: Industrial Biochemistry Programme, Department of Chemical & Environmental Sciences, University of Limerick, Limerick city, Ireland.
authors.forename: Anne
authors.initials: A
authors.lastname: Casey
authors.forename: Gary
authors.initials: G
authors.lastname: Walsh
mesh_terms: D010832:6-Phytase / Q000302:isolation & purification* / Q000494:pharmacology; D000821:Animal Feed; D001234:Aspergillus niger / Q000201:enzymology*; D002852:Chromatography, Ion Exchange; D005374:Filtration; D019216:Metals, Heavy
chemical_list: D019216:Metals, Heavy; D010832:6-Phytase