Patent ID: 7425436

Claim:
A polynucleotide comprising a nucleic acid sequence encoding a mutant of the dehalogenase of SEQ ID NO: 82, the mutant having at least 85% sequence identity to SEQ ID NO: 82, and the mutant comprising at least two amino acid substitutions, wherein the mutant dehalogenase forms a bond with a dehalogenase substrate which comprises one or more functional groups, which bond is more stable than the bond formed between the dehalogenase of SEQ ID NO: 82 and the substrate, wherein at least one amino acid substitution in the mutant dehalogenase is a substitution at an amino acid residue corresponding to residue 272 of SEQ ID NO:82, a residue that is associated with activating a water molecule which cleaves the bond formed between the corresponding wild-type dehalogenase and the substrate, and wherein a second substitution is at an amino acid residue corresponding to residue 175 in the Rhodococcus dehalogenase having the amino acid sequence of SEQ ID NO:82.