Purification and characterisation of an acid phosphatase with phytase activity from Mucor hiemalis Wehmer.
An acid phosphatase with phytase activity, produced by Mucor hiemalis Wehmer, was purified to homogeneity by a combination of anion exchange, gel filtration and hydrophobic interaction chromatography. The monomeric, glycosylated enzyme displayed maximum activity at 55 degrees C and pH 5.0-5.5. When compared to commercialised products, the enzyme is more thermostable (80 degrees C, 5min), displays a broader pH versus activity profile and greater stability under simulated digestive tract conditions. Unlike commercial phytases, the Mucor enzyme should retain some activity in the small intestine as well as in the stomach, facilitating a longer duration of action and hence more extensive substrate hydrolysis. Substrate specificity studies and protein database similarity searching using mass spectrometry-derived sequence data indicate that the enzyme is an acid phosphatase with activity on phytate. Cocktails containing acid phosphatases in combination with true phytases have been shown to promote more extensive phytate degradation than do true phytases alone. This, coupled to the enzyme's functionally relevant physicochemical characteristics, suggests its likely suitability for inclusion in second generation phytase cocktails for application in animal feed.
Boyce
A
A
Department of Chemical and Environmental Sciences, University of Limerick, Limerick City, Ireland.
Walsh
G
G
D010832:6-Phytase / Q000008:administration & dosage / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D000135:Acid Phosphatase / Q000008:administration & dosage / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D000595:Amino Acid Sequence; D000821:Animal Feed; D000818:Animals; D001709:Biotechnology; D004064:Digestive System / Q000201:enzymology; D004795:Enzyme Stability; D005503:Food Additives; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009089:Mucor / Q000201:enzymology* / Q000235:genetics; D013379:Substrate Specificity; D034421:Sus scrofa; D013696:Temperature
D005503:Food Additives; D000135:Acid Phosphatase; D010832:6-Phytase