Deciphering reductive dehalogenase specificity through targeted mutagenesis of chloroalkane reductases.

ABSTRACT: Reductive dehalogenases (RDases) are essential in the anaerobic degradation of various organohalide contaminants. This family of enzymes has broad sequence diversity, but high structural conservation. There have been few studies assessing how RDase amino acid sequences affect their substrate selectivity. Here, we focus on two chloroalkane RDases, CfrA and DcrA, which have 95% protein sequence identity but have diverged to have opposite substrate preferences. CfrA dechlorinates chloroform (CF) and 1,1,1-trichloroethane (TCA) but not 1,1-dichloroethane (DCA), while DcrA will dechlorinate 1,1-DCA but not CF or 1,1,1-TCA. We mutated several residues in the active site of CfrA to investigate a change in substrate preference and to identify which wild-type residues contribute the most to substra

SUBMITTER: Picott KJ 

PROVIDER: S-EPMC11921367 | biostudies-literature | 2025 Mar

REPOSITORIES: biostudies-literature