Altered thermostability of enzymes
AssigneeDiversa Corporation
InventorJay M. Short
Life of this patent
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Abstract
Provided is a method of screening gene libraries derived from a mixed population of organisms for a bioactivity or biomolecule of interest. The mixed population of organisms can be a cultured population or an uncultured population from, for example, the environment. Also provided are methods of screening isolates or enriched populations of organisms, which isolates include a population that is spatially, temporally, or hierarchical, for example, of a particular species, genus, family, or class of organisms. Identified clones containing a biomolecule or bioactivity of interest can be further variegated or the DNA contained in the clone can be variegated to create novel biomolecules or bioactivities of interest.
Claims61
A method for identifying a protein or polypeptide having an altered thermostability compared with a thermostability of a wild-type protein or polypeptide, the method comprising:
- a) constructing a library of clones, wherein the clones are constructed from nucleic acids obtained from an unselected mixed population of organisms from an environmental sample;
- (b) determining thermostability of one or more wild-type proteins or polypeptides expressed from one or more clones of the library;
- (c) subjecting one or more of the clones of (b) to mutagenesis;
- (d) determining thermostability of modified proteins or polypeptides expressed from the clones of (c);
- (e) comparing the thermostability of the one or more wild-type proteins or polypeptides of (b) with the thermostability of at least one corresponding modified protein or polypeptide of (d), thereby identifying one or more clones expressing a modified proteins or polypeptides having an altered thermostability, and;
- (f) selecting at least one protein or polypeptide having an altered thermostability.
Dependencies are inferred from the claim text.