Abstract
Expression of nitrile hydratase enzymes utilized in a new “green” process for acrylamide production has proven difficult in Escherichia coli owing to lack of a cobalt transport system to introduce the required cobaltion into this host. We describe the expression of a thermostable nitrile hydratase from a moderatethermophile Bacillus sp. BR449 in E. coli in which the cobaltrequired for enzyme activation is introduced by incubation, of the apoenzyme in the presence of Co++ ion at 50°C, yielding active and thermostable, enzyme.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ashina, Y. and Suto, M. (1993), in Industrial Applications of Immobilized Biocatalysts, Tanaka, A., Tosa, T., and Kobayashi, T., eds., Marcel Dekker, New York, pp. 91–107.
Yamada, H. and Kobayashi, M. (1996), Biosci. Biotech. Biochem. 60, 1391–1400.
Kobayashi, M. and Shimizu, S. (1998), Nat. Biotechnol. 16, 733–736.
Ikehata, O., Nishiyama, M., Horinoushi, S., and Beppu, T. (1989), J. Biochem. 181, 563–570.
Kobayashi, M., Snishiyama, M., Nagasawa, T., Horinoushi, S., Beppu, T., and Yamaday, H. (1991), Biochim. Biophys. Acta 1129, 23–33.
Wu, S., Fallon, R. D., and Payne, M. S., (1997), Appl. Microbiol. Biotechnol. 48, 704–708.
Nojiri, M., Yohda, M., Masafumi, O., et al. (1999), J. Biochem. 125, 696–704.
Kim, S. H. and Oriel, P. (2000), Enz. Microb. Technol. 27, 492–501.
Padmakumar, R. and Oriel, P., (1999), Appl. Biochem. Biotechnol. 77, 671–680.
Payne, M. S., Wu, S., Fallon, R. D., Tudor, G., Stieglitz, B., Turner, I. M., Jr., and Nelson, M. J. (1997), Biochemistry 36, 5447–5454.
Nagasawa, T., Takeuchi, K., and Yamada, H., (1991), Eur. J. Biochem. 196, 581–589.
Brennan, B. A., Alms, G., Nelson, M. J., Durney, L. T., and Scarrow, R. C., (1996), J. Am. Chem. Soc. 118, 9194–9195.
Nojiri, M., Nakayama, H., Odaka, M., Yohda, M., Takio, K., and Endo, I., (2000), FEBS Lett. 465, 173–177.
Friedman, M., Calvins, J. F., and Wall, J. S., (1965), J. Am. Chem. Soc. 87, 3672–3682.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kim, SH., Padmakumar, R. & Oriel, P. Cobalt activation of Bacillus BR449 thermostable nitrile hydratase expressed in Escherichia coli . Appl Biochem Biotechnol 91, 597–603 (2001). https://doi.org/10.1385/ABAB:91-93:1-9:597
Issue Date:
DOI: https://doi.org/10.1385/ABAB:91-93:1-9:597