Mechanistic Dichotomy in Bacterial Trichloroethene Dechlorination Revealed by Carbon and Chlorine Isotope Effects

Tetrachloroethene (PCE) and trichloroethene (TCE) are significant groundwater contaminants. Microbial reductive dehalogenation at contaminated sites can produce nontoxic ethene but often stops at toxic cis-1,2-dichloroethene (cis-DCE) or vinyl chloride (VC). The magnitude of carbon relative to chlorine isotope effects (as expressed by Λ_C/Cl, the slope of δ¹³C versus δ³⁷Cl regressions) was recently recognized to reveal different reduction mechanisms with vitamin B₁₂ as a model reactant for reductive dehalogenase activity. Large Λ_C/Cl values for cis-DCE reflected cob­(I)­alamin addition followed by protonation, whereas smaller Λ_C/Cl values for PCE evidenced cob­(I)­alamin addition followed by Cl^– elimination. This study addressed dehalogenation in actual microorganisms and observed identical large Λ_C/Cl values for cis-DCE (Λ_C/Cl = 10.0 to 17.8) that contrasted with identical smaller Λ_C/Cl for TCE and PCE (Λ_C/Cl = 2.3 to 3.8). For TCE, the trend of small Λ_C/Cl could even be reversed when mixed cultures were precultivated on VC or DCEs and subsequently confronted with TCE (Λ_C/Cl = 9.0 to 18.2). This observation provides explicit evidence that substrate adaptation must have selected for reductive dehalogenases with different mechanistic motifs. The patterns of Λ_C/Cl are consistent with practically all studies published to date, while the difference in reaction mechanisms offers a potential answer to the long-standing question of why bioremediation frequently stalls at cis-DCE.