Identification and characterization of a phytase of potential commercial interest.
Phytases catalyse the hydrolytic degradation of phytic acid and its salts and are added to monogastric animal feed to ameliorate the negative environmental and nutritional consequences of dietary phytate. Screening of 58 microbial strains identified a phytase produced by Rhizopus oligosporus ATCC 22959 that displayed physicochemical characteristics likely to render it of potential industrial interest. The 124 kDa enzyme was purified to homogeneity by anion exchange chromatography, gel filtration and chromatofocusing. The monomeric glycosylated enzyme (30.5% total carbohydrate) displayed maximum activity at 65 degrees C and pH 5.0. It displayed a Km of 10.4 microM, a Vmax of 1.32 nmols(-1) and a Kcat of 51 s(-1). It is acid tolerant, retaining full activity after incubation at pH 2.0 for 6h. HPLC analysis indicated the enzyme's ability to almost completely degrade phytate. Substrate specificity studies showed its ability to dephosphorylate several additional phosphorylated molecules. Activity was unaffected or moderately stimulated by a range of metal ions with only Ca2+ exerting a modest (13%) inhibitory effect. The enzyme is significantly more thermostable at 80 degrees C and retains a significantly greater proportion of maximal activity at physiological temperatures than do two commercial phytases tested for comparative purposes. This may render it of industrial interest.
Casey
Anne
A
Industrial Biochemistry Program, Department of Chemical & Environmental Sciences, University of Limerick, Limerick City, Ireland.
Walsh
Gary
G
D010832:6-Phytase / Q000737:chemistry* / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism; D000821:Animal Feed; D001426:Bacterial Proteins / Q000737:chemistry* / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism; D002118:Calcium / Q000494:pharmacology; D002241:Carbohydrates / Q000737:chemistry; D002384:Catalysis; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004795:Enzyme Stability; D005374:Filtration; D006031:Glycosylation; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D010766:Phosphorylation; D012233:Rhizopus / Q000201:enzymology; D013379:Substrate Specificity; D013696:Temperature; D013997:Time Factors
D001426:Bacterial Proteins; D002241:Carbohydrates; D004791:Enzyme Inhibitors; D010832:6-Phytase; D002118:Calcium