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US 5,830,696 AGranted
Directed evolution of thermophilic enzymes
AssigneeDiversa Corporation
InventorJay M. Short
Life of this patent
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Abstract
Thermostable enzyme are subjected to mutagenesis to produce a thermophilic enzyme which is stable at thermophilic temperature and which has increased activities at least two-fold higher than the activity of the wild-type thermostable enzyme at lower temperatures, which are temperatures of 50.degree. C. or lower.
Claims2
A method for providing a thermostable enzyme having improved enzyme activities as compared to a corresponding wild-type enzyme at lower temperatures, comprising:
- (a) subjecting at least one polynucleotide encoding an enzyme which is stable at a temperature of at least 60.degree. C. to a random mutagenesis;
- and
- (b) screening mutant enzymes produced in (a) for a mutated enzyme or for a polynucleotide encoding a mutated enzyme, wherein the mutated enzyme is stable at a temperature of at least 60.degree. C.;
- has an enzyme activity at a temperature at least 10.degree. C. below its optimal temperature range;
- and has activity greater than the enzyme encoded by the polynucleotide of step (a) prior to mutagenesis.
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