Isolation and characterization of a novel phytase from Penicillium simplicissimum.
Eighty-three isolates from different soil samples exhibited the potential for producing active extracellular phytase. The most active fungal isolate with phytase activity was identified as Penicillium simplicissimum. In shaking culture with enrichment medium, the highest extracellular phytase activity of the producing strain was 3.8 U/mL. The crude enzyme filtrate was purified to homogeneity using ultrafiltration. IEC and gel filtration chromatography. The molar mass of the purified enzyme was estimated to be 65 kDa on SDS-PAGE. The saccharide identification with periodic acid-Schiff reagent (PAS) and activity recognition by 1-naphthyl phosphate was all positive. The isoelectric point of the enzyme, as deduced by isoelectric focusing, was pH 5.8, the optimum pH and temperature being pH 4.0 and 55 degrees C, respectively. The purified enzyme revealed broad substrate specificity and was strongly inhibited by Fe2+, Fe3+ and Zn2+; however, no inhibition was found by EDTA and PMSF. Phytase activity was inhibited when 2 mmol/L of dodecasodium phytate was added and the Km for it was determined to be 813 mmol/L.
Department of Food Science, National Chung Hsing University, Taichung, Taiwan 40227, Republic of China.
Y H
YH
Tseng
T J
TJ
Fang
S M
SM
Tseng
D010832:6-Phytase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007477:Ions / Q000494:pharmacology; D008670:Metals / Q000494:pharmacology; D008970:Molecular Weight; D010407:Penicillium / Q000201:enzymology*; D012988:Soil Microbiology; D013379:Substrate Specificity; D013696:Temperature; D014462:Ultrafiltration
D007477:Ions; D008670:Metals; D010832:6-Phytase