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Science 6 May 2005:
Vol. 308. no. 5723, pp. 857 - 860
DOI: 10.1126/science.1107387
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Reports

Computational Thermostabilization of an Enzyme

Aaron Korkegian,1,2 Margaret E. Black,4 David Baker,3 Barry L. Stoddard1*

Thermostabilizing an enzyme while maintaining its activity for industrial or biomedical applications can be difficult with traditional selection methods. We describe a rapid computational approach that identified three mutations within a model enzyme that produced a 10°C increase in apparent melting temperature Tm and a 30-fold increase in half-life at 50°C, with no reduction in catalytic efficiency. The effects of the mutations were synergistic, giving an increase in excess of the sum of their individual effects. The redesigned enzyme induced an increased, temperature-dependent bacterial growth rate under conditions that required its activity, thereby coupling molecular and metabolic engineering.

1 Division of Basic Sciences, Fred Hutchinson Cancer Research Center (FHCRC), 1100 Fairview Avenue North, Seattle, WA 98109, USA.
2 Graduate Program in Molecular and Cellular Biology, University of Washington, Seattle, WA 98195, USA.
3 Department of Biochemistry and Howard Hughes Medical Institute, University of Washington, Seattle, WA 98195, USA.
4 Department of Pharmaceutical Sciences, College of Pharmacy, Washington State University, Pullman, WA 99164–6534, USA.

* To whom correspondence should be addressed. E-mail: bstoddar{at}fhcrc.org

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