Abstract
A lactate oxidase was purified about 36-fold from a newly screened strain KY6 of gram negative bacterium from soil to yield a homogeneous protein. The native enzyme had a molecular mass of 204 kDa measured by Sephadex G-200 and that of subunit on the SDS-PAGE was found to be 45 kDa. The enzyme was optimally active at pH 7.7 and showed stability at pH range of 5.7 to 9.5 for 24 h at 4‡C. The optimum temperature was 70‡C and the enzyme activity was stable for 10 min up to 45‡C. The half-life of the enzyme activity was about 10 min at 55‡C. The best substrate of the enzyme was D-lactate and Km value for D-lactate was 0.14 mM. The Km value for DL-lactate was 0.20 mM. Substrate inhibition of the enzyme was observed at higher concentrations than 20 mM of DL-lactate and 10 mM of D-lactate.
Similar content being viewed by others
References
Yanase, H., Mori, N., Masuda, M., Kita, K., Shimao, M., and Kato, N. (1992),J. Ferment. Bioeng. 73, 287–291.
Yamamura, Y., Kusunose, M., and Kusunose, E. (1952),Nature 170, 207–208.
Xu, P., Yano, T., Yamamoto, K., Suzuki, H., and Kumagai, H. (1996),J. Ferment. Bioeng. (in press).
Robinson, J. C., Keay, L., Molinari, R., and Sizer, I. W. (1962),J. Biol. Chem. 237, 2001–2010.
Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. (1951),J. Biol. Chem. 193, 265–275.
Laemmli, U. K. (1970),Nature 227, 680–685.
Horecker, B. L. (1950),J. Biol. Chem. 183, 593–605.
Thanos, I., Bader, J., Gunther, H., Neumann, S., Krauss, F., and Simon, H. (1987),Methods in Enzymology 136, 302–317.
Sutton, W. B. (1954),J. Biol. Chem. 210, 210–320.
Sneath, P. H. A., Mair, N. S., Sharpe, M. E., and Polt, J. G. (1986), “Bergey’s Manual of Systematic Bacteriology”2, 1436.
Tokushige, M. and Sizer, W. (1967),J. Biochem. 62, 719–725.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Xu, P., Yano, T., Yamamoto, K. et al. Characterization of a lactate oxidase from a strain of gram negative bacterium from soil. Appl Biochem Biotechnol 56, 277–288 (1996). https://doi.org/10.1007/BF02786958
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02786958