Abstract
Bacillus species producing a thermostable phytase was isolated from soil, boiled rice, and mezu (Korean traditinal koji). The activity of phytase increased markedly at the late stationary phase. An extracellular phytase from Bacillus sp. KHU-10 was purified to homogeneity by acetone precipitation and DEAE-Sepharose and phenyl-Sepharose column chromatographies. Its molecular weight was estimated to be 46 kDa on gel filtration and 44 kDa on SDS-polyacrylamide gel elctrophoresis. Its optimum pH and temperature for phytase activity were pH 6.5-8.5 and 40°C without 10 mM CaCl2 and pH 6.0-9.5 and 60°C with 10 mM CaCl2. About 50% of its original activity remained after incubation at 80°C or 10 min in the presence of 10 mM CaCl2. The enzyme activity was fairly stable from pH 6.5 to 10.0. The enzyme had an isoelectric point of 6.8. As for substrate specificity, it was very specific for sodium phytate and showed no activity on other phosphate esters. The K m value for sodium phytate was 50 μM. Its activity was inhibited by EDTA and metal ions such as Ba2+, Cd2+, Co2+, Cr3+, Cu2+, Hg2+, and Mn2+ ions.
Similar content being viewed by others
REFERENCES
Ahmad, T., Rasool, S., Sarwar, M., Haq, A. U., and Hasan, Z. U. (2000). Anim.Feed Sci.Tech. 83, 103–114.
Choi, Y. M., Noh, D. o., Cho, S. H., Lee, H. K., Suh, H. J., and Chung, S. H. (1999). J.Microbiol.Biotech. 9, 223–226.
Dvorakova, J., Volfova, O., and Kopecky, J. (1997). Folia Microbiol. 42, 349–352.
Gibson, D. M. and Ullah, A. H. J. (1988). Arch.Biochem.Biophys. 260, 503–513.
Greiner, R., Konietzny, U., and Jany, K. D. (1993). Arch.Biochem.Biophys. 303, 107–113.
Greiner, R., Haller, E., Konietzny, U., and Jany, K. D. (1997). Arch.Biochem.Biophy. 341, 201–206.
Han, Y. W. and Gallagher, D. J. (1987). J.Ind.Microbiol. 2, 295–301.
Irving, G. C. J. (1980). In Inositol Phosphates: Their Chemistry, Biochemistry, and Physiology (Cosgrove, D. J., ed.), Elsevier Press, Amsterdam, pp. 85–98.
Kerovuo, J., Lauraeus, M., Nurminen, P., Kalkkinen, N., and Apajalahti, J. (1998). Appl.Environ.Microbiol. 64, 2079–2085.
Kim, D. S., Godber, J. S., and Kim, H. R. (1999). Biotechnol.Lett. 21, 1077–1081.
Kim, Y. O., Kim, H. K., Bae, K. S., Yu, J. H., and T. K. (1998). Enzyme Microbiol.Technol. 22, 2–7.
Konietzny, U., Greiner, R., and Jany, K. (1995). J.Food Biochem. 18, 165–183.
Martinez, C., Ros, G., Periago, M. J., Lopez, G., Ortuno, J., and Rincon, G. (1996). Food Sci.Technol.Int. 2, 201–209.
Mayer, A. F., Hellmuth, K., Schlieker, H., Lopez-Ulibarri, R., Oertel, S., Dahlems, U., Strasser, A. W. M., and van Loon, A. P. G. M. (1999). Biotechnol.Bioeng. 63, 373–381.
Nayini, N. R. and Markakis, P. (1986). In Phytic acid; Chemistry and Applications (Graf, E., ed.), Pilatus Press, Mineapolis, MN, pp. 101–118.
Pandey, A., Szakacs, G., Soccol, C. R., Rodriguez-Leon, J. A., and Soccol, V. T. (2001). Bioresource Technol. 77, 203–214.
Powar, V. K. and Jagannathan, V. (1982). J.Bacteriol. 151, 1102–1108.
Shimizu, M. (1992). Biosci.Biotechnol.Biochem. 56, 1266–1269.
Sreeramulu, G., Srinivasa, D. S., Nand, K., and Joseph, R. (1996). Lett.Appl.Microbiol. 23, 385–388.
Sztajer, H., Lnsdorf, H., Erdmann, H., Menge, U., and Schmid, R. (1992). Biochim.Biophys.Acta 1124, 253–261.
Ullah, A. H. J. (1988). Prep.Biochem. 18, 459–471.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Choi, Y.M., Suh, H.J. & Kim, J.M. Purification and Properties of Extracellular Phytase from Bacillus sp. KHU-10. J Protein Chem 20, 287–292 (2001). https://doi.org/10.1023/A:1010945416862
Published:
Issue Date:
DOI: https://doi.org/10.1023/A:1010945416862