Purification and kinetics of a protease-resistant,neutral, and thermostable phytase from Bacillus subtilis subsp. subtilis JJBS250 ameliorating food nutrition |
| |
Authors: | Jinender Jain Anil Kumar Davender Singh |
| |
Affiliation: | 1. Department of Microbiology, Laboratory of Bioprocess Technology, Maharshi Dayanand University, Rohtak, Haryana, India;2. Department of Botany, Pt. Neki Ram Sharma Government College, Rohtak, Haryana, India;3. Department of Physics, RPS Degree College, Balana, Haryana, India |
| |
Abstract: | AbstractA novel protease-resistant and thermostable phytase from Bacillus subtilis subsp. subtilis JJBS250 was purified 36-fold to homogeneity with a combination of ammonium sulfate precipitation followed by Q-Sepharose and Sephadex G-50 chromatographic techniques. The estimated molecular mass of the purified phytase was 46?kDa by electrophoresis with optimal activity at pH 7.0 and 70?°C. About 19% of original activity was maintained at 80?°C for 10?min. Phytase activity was stimulated in presence of surfactants like Tween-20, Tween-80, and Triton X-100 and metal ions like Ca+2, K+, and Co+2 and it was inhibited by SDS and Mg+2, Al+2, and Fe+2. Purified enzyme showed specificity to different salts of phytic acid and values of Km and Vmax were 0.293?mM and 11.49 nmoles s?1, respectively for sodium phytate. The purified enzyme was resistant to proteases (trypsin and pepsin) that resulted in amelioration of food nutrition with simultaneous release of inorganic phosphate, reducing sugars, and soluble protein. |
| |
Keywords: | Bacillus subtilis subsp. subtilis JJBS250 dephytinization phytase purification protease-resistant |
|
|