Investigating mechanisms of chlorinated ethane biotransformation using compound specific carbon isotope analysis.

摘要

Stable carbon isotope fractionation during biotransformation of the chlorinated ethanes 1,2-dichloroethane (1,2-DCA), 1,1,1-trichloroethane (1,1,1-TCA) and 1,1-dichloroethane (1,1-DCA) was investigated. Isotopic fractionation during aerobic 1,2-DCA biotransformation in microcosms, enrichment cultures and pure microbial cultures was measured and was found to be pathway dependent. Biodegradation of 1,2-DCA under aerobic conditions produced a consistent bimodal distribution of enrichment factors (s) with one mean c centered on --3.9 +/- 0.6‰ and the other on --29.2 +/- 1.9‰. Reevaluation of epsilon in terms of kinetic isotope effects 12k/ 13k, gave values of 12k/13k = 1.01 and 1.06, which are typical of oxidation and hydrolytic dehalogenation (S N2) reactions, respectively. The relationship between degradation pathway and measured carbon isotope fractionation was applied to constrain the degradation pathway of 1,2-DCA in a microbial enrichment culture capable of degrading 1,2-DCA under both O2 and NO3-reducing conditions, but where the degradation pathway was previously uncharacterized. delta 13C values indicated biodegradation in the enrichment culture under both O2 and NO3-reducing conditions likely proceeded via a hydrolytic dehalogenase enzyme. Stable carbon isotope analysis during biotransformation of other chlorinated ethanes was then investigated. Isotopic enrichment factors of -1.9‰ and -10.4‰ were measured during reductive dechlorination of 1,1,1-trichloroethane (1,1,1-TCA) and 1,1-dichloroethane (1,1-DCA), respectively. These are the first reported isotopic enrichment factors for microbial biotransformation of these compounds, which can now potentially be applied to investigate and quantify biodegradation of 1,1,1-TCA and 1,1-DCA in the field. Stable carbon isotope analysis was used to provide a conservative estimate of the extent of 1,2-DCA and trichloroethene (TCE) biodegradation in a biostimulation field study. Isotope analysis was able to confirm that 1,2-DCA degradation, rather than degradation of vinyl chloride, was the primary mechanism of ethene production at the site. This thesis not only advances the application of compound specific carbon isotope fractionation to identify and quantify biodegradation of chlorinated ethanes in groundwater, but in particular progresses our understanding of the relationship between the enzymatic mechanisms of contaminant degradation and isotopic fractionation, and the ability to use that knowledge to predict degradation mechanisms for previously unconstrained pathways of contaminant remediation.