Mutant hydrolase proteins with enhanced kinetics and functional expression
AssigneePromega Corporation
InventorsAldis Darzins, Lance P. Encell, Rachel Friedman Ohana, Paul Otto, Michael R. Slater, Gediminas Vidugiris, Keith V. Wood, Monika G. Wood, Kate Qin Zhao, Kris Zimmerman
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Abstract
The invention provides a mutant hydrolase protein with enhanced kinetics and functional expression, as well as polynucleotides encoding the mutant proteins and methods of using the polynucleotides and mutant proteins.
Claims6
A polypeptide comprising a mutant dehalogenase that is capable of catalyzing hydrolytic dehalogenation of an alkylhalide substrate and forming a covalent ester intermediate with the substrate, said mutant dehalogenase having at least 85% sequence identity with the polypeptide of SEQ ID NO:
- 1;
- wherein said mutant dehalogenase comprises substitutions at:
- (a) one or more positions corresponding to positions in SEQ ID NO:
- 1 selected from the group consisting of positions 106 and 272;
- and
- (b) positions corresponding to positions 58, 78, 155, 167, 172, 224, and 291 of SEQ ID NO:
- 1.
Dependencies are inferred from the claim text.