Computational thermostabilization of an enzyme
|Title||Computational thermostabilization of an enzyme|
|Publication Type||Journal Article|
|Year of Publication||2005|
|Authors||Korkegian, A., Black M. E., Baker D., & Stoddard B. L.|
|Date Published||2005 May 6|
|Keywords||Amino Acid Sequence, Binding Sites, Catalysis, Circular Dichroism, Collaborative Publication, Computer Simulation, Crystallography, X-Ray, Cytosine Deaminase, Enzyme Stability, Escherichia coli, Kinetics, Models, Molecular, Molecular Sequence Data, Monte Carlo Method, Mutagenesis, Site-Directed, Point Mutation, Protein Conformation, Protein Denaturation, Protein Engineering, Protein Folding, Protein Structure, Secondary, Software, Temperature, Thermodynamics, Transformation, Genetic, Yeasts|
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 degrees C increase in apparent melting temperature T(m) and a 30-fold increase in half-life at 50 degrees 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.