The BhbA Enzyme Assay

Abstract

Reductive dehalogenation has been found primarily in anaerobic communities and is originally thought to rarely occur in aerobes. A reductive dehalogenase (BhbA) was characterized from an aerobic strain of Comamonas sp. 7D-2, which was isolated from a bromoxynil octanoate-contaminated soil sample collected in Jiangsu, China. BhbA catalyzes the reductive dehalogenation of bromoxynil and its derivative 3,5-dibromo-4-hydroxybenzoate under aerobic conditions. BhbA is membrane-associated and found to have the key features of anaerobic respiratory reductive dehalogenases. This protocol describes the method for enzyme analysis of the aerobic reductive dehalogenase (BhbA) in the membrane fraction.