Phytase is enzyme which catalyze the hydrolysis of phytic acid into myo-inositol and inorganic phosphate. This study was carried out to isolate the phytase producing microorganisms from natural sources. The isolate CF 92 which was isolated from cattle feces showed the highest phytase activity. It wa...
Phytase is enzyme which catalyze the hydrolysis of phytic acid into myo-inositol and inorganic phosphate. This study was carried out to isolate the phytase producing microorganisms from natural sources. The isolate CF 92 which was isolated from cattle feces showed the highest phytase activity. It was identified as Bacillus subtilis, and designated as Bacillus subtilis CF 92. The production of phytase from Bacillus subtilis CF 92 reached the high level when grown at 37 ℃ for 48 h in PSL medium. The optimal medium compositions for enzyme production were 10 %(w/v) rice bran extract, 0.05 %(w/v) whey protein powder, 0.02 %(w/v) calcium chloride dihydrate and 0.02 %(w/v) potassium dihydrogenphosphate. The phytase was purified by ethanol precipitation, ion-exchange chromatography, and gel filtration. The purified phytase showed the optimum temperature of 60 ℃ and 40 % of its original activity remained when it treated at 80 ℃ for 30 min. Its optimum pH was 7.0 and fairly stable from pH 4.0 - 8.0. The enzyme activity was activated in the presence of EDTA, even though it was inhibited in the presence of metal-ions. As regards substrate specificity, the phytase had specificity for polyphosphate compounds such as adenosine triphosphate, sodium tripolyphosphate, and sodium phytate. The Km and Vmax value for the sodium phytate was 0.42 mM and 4.35 μmole/min, respectively.
Phytase is enzyme which catalyze the hydrolysis of phytic acid into myo-inositol and inorganic phosphate. This study was carried out to isolate the phytase producing microorganisms from natural sources. The isolate CF 92 which was isolated from cattle feces showed the highest phytase activity. It was identified as Bacillus subtilis, and designated as Bacillus subtilis CF 92. The production of phytase from Bacillus subtilis CF 92 reached the high level when grown at 37 ℃ for 48 h in PSL medium. The optimal medium compositions for enzyme production were 10 %(w/v) rice bran extract, 0.05 %(w/v) whey protein powder, 0.02 %(w/v) calcium chloride dihydrate and 0.02 %(w/v) potassium dihydrogenphosphate. The phytase was purified by ethanol precipitation, ion-exchange chromatography, and gel filtration. The purified phytase showed the optimum temperature of 60 ℃ and 40 % of its original activity remained when it treated at 80 ℃ for 30 min. Its optimum pH was 7.0 and fairly stable from pH 4.0 - 8.0. The enzyme activity was activated in the presence of EDTA, even though it was inhibited in the presence of metal-ions. As regards substrate specificity, the phytase had specificity for polyphosphate compounds such as adenosine triphosphate, sodium tripolyphosphate, and sodium phytate. The Km and Vmax value for the sodium phytate was 0.42 mM and 4.35 μmole/min, respectively.
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