Diversity of alpha-halocarboxylic acid dehalogenases in bacteria isolated from a pristine soil after enrichment and selection on the herbicide 2,2-dichloropropionic acid (Dalapon)

Five pure cultures of bacteria (strains DA1–5) able to degrade 2,2-dichloropropionic acid (22DCPA) were isolated for the first time from pristine bulk soil samples. From 16S rDNA analysis, it was concluded that strains DA2, DA3 and DA4 were members of the Bradyrhizobium subgroup (α-Proteobacteria), strain DA5 clustered in the Brucella assemblage (α-Proteobacteria) and strain DA1 clustered in the β-Proteobacteria. Biochemical and molecular analysis of the dehalogenases from the isolates showed that these enzymes were quite diverse. Several dehalogenases were closely related to group I and II α-halocarboxylic acid dehalogenases, and partial polymerase chain reaction (PCR) products were obtained from isolates DA1, 2, 3 and 4 using degenerate dehalogenase primers. However, no PCR products were obtained from isolate DA5 using either of the group I or II α-halocarboxylic acid dehalogenase primers. Isolates DA2 and DA4 contained putative silent dehalogenases. The investigation highlighted the endemic nature of these genes in pristine environments and how diverse these were even from spatially close samples