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Engineering Enzymes for Enhanced Thermostability

Published online by Cambridge University Press:  15 February 2011

Phoebe Shih ,
Bruce A. Malcolm  and
Jack F. Kirsch
Phoebe Shih
Affiliation:
Department of Molecular and Cellular Biology, Barker Hall, University of California, Berkeley, CA 94720 and Center for Advanced Materials, Lawrence Berkeley Laboratory, Berkeley, CA 94720
Bruce A. Malcolm
Affiliation:
Department of Molecular and Cellular Biology, Barker Hall, University of California, Berkeley, CA 94720 and Center for Advanced Materials, Lawrence Berkeley Laboratory, Berkeley, CA 94720
Jack F. Kirsch
Affiliation:
Department of Molecular and Cellular Biology, Barker Hall, University of California, Berkeley, CA 94720 and Center for Advanced Materials, Lawrence Berkeley Laboratory, Berkeley, CA 94720
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Abstract

Chicken egg-white lysozyme (CEWL) is used as a model to attempt to engineer proteins for enhanced thermostability. Site-directed mutagenesis is employed for selective amino acid substitution to probe the contribution of an individual amino acid in a given sequence to thermostability. A linear correlation is found between the side-chain volume of a triplet of amino acid residues located at the interior core of the protein and its thermostability. Additional mutant constructs at the core position reveal that hyperpacking can disrupt other intramolecular contacts and offset the hydrophobic stabilization due to denser packing. Multiple substitutions at different loci of the protein are combined to analyze the additivity of thermostability mutations.

Type
Research Article
Information
MRS Online Proceedings Library (OPL) , Volume 218: Symposium V – Materials Synthesis Based on Biological Processes , 1990 , 37
Copyright
Copyright © Materials Research Society 1991

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References

REFERENCES

1. Stearman, R. S., Frankel, A. D., Freire, E., Liu, B., and Pabo, C. O., Biochemistry, 27, 7571 (1988).Google Scholar
2. Matthews, B. W., Nicholson, H., and Becktel, W. J., Proc. Nad. Acad. Sci. USA, 84, 6663 (1987).Google Scholar
4. Perry, L. J. and Wetzel, R., Science, 226, 555 (1984).Google Scholar
5. Foss, J., Biochim. Biophys. Acta, 47, 569 (1961).Google Scholar
6. O'Reilly, J. R. and Karasz, F. E., Biopolymers, 9, 1429 (1970).Google Scholar
7. Malcolm, B. A., Ph. D. Dissertation, University of California Berkeley, (1988).Google Scholar
8. Malcolm, B. A., Wilson, K. P., Matthews, B. W., Kirsch, J. F., and Wilson, A. C., Nature, 345, 86 (1990).Google Scholar
9. Malcolm, B. A., Rosenberg, S., Corey, M. J., Allen, J. S., Baetselier, A. de, and Kirsch, J. F., Proc. Natl. Acad. Sci. USA, 35, 1533 (1988).Google Scholar
10. Chothia, C., Nature, 254, 304 (1975).Google Scholar
11. Matsumura, M., Becktel, W. J., and Matthews, B. W., Nature, 334, 406 (1988).Google Scholar
12. Karpusas, M., Baase, W. A., and Matthews, B. W., Proc. Natl. Acad. Sci. U.S.A., 86, 8237 (1989).CrossRefGoogle Scholar
13. Sandberg, W. S. and Terwilliger, T. C., Science, 245, 54 (1989).Google Scholar
14. Kirsch, J. F., Malcolm, B. A., Corey, M. J., Zhang, L., and Wilson, A. in The Immune Response to Structurally Defined Proteins: The Lysozyme Model, edited by Smith-Gill, S. and Sercarz, E. (Adenine Press, Schenectady, New York, 1989), pp. 59–63.Google Scholar
16. Wells, J. A., Biochemistry, 29, 8509 (1990).Google Scholar