Field and in vitro evidence for in-situ bioremediation using compound-specific 13C/12C ratio monitoring

Stehmeier, L.G.; Francis, Mike McD.; Jack, Thomas; Diegor, E. J. M.; Winsor, Linda; Abrajano, T.A.

doi:10.1016/s0146-6380(99)00065-0

Cited by 55 publications

(40 citation statements)

References 13 publications

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“…and Burkholderia sp. Others observed a small amount of 13 C/ 12 C isotope fractionation during benzene degradation by an aerobic mixed culture that was enriched from groundwater of a petrochemical site (27). The small isotope fractionation factors and the increase in ␦ 13 C for the initial mono-or dioxygenase reactions at the benzene ring that were documented in these papers are in agreement with our observations that oxygenases acting on ⌸-electron system of the aromatic ring produce only minor isotope effects.…”

supporting

confidence: 91%

Carbon and Hydrogen Stable Isotope Fractionation during Aerobic Bacterial Degradation of Aromatic Hydrocarbons

Morasch

Richnow

Schink

et al. 2002

Appl Environ Microbiol

121

109

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13 C/ 12 C and D/H stable isotope fractionation during aerobic degradation was determined for Pseudomonas putida strain mt-2, Pseudomonas putida strain F1, Ralstonia pickettii strain PKO1, and Pseudomonas putida strain NCIB 9816 grown with toluene, xylenes, and naphthalene. Different types of initial reactions used by the respective bacterial strains could be linked with certain extents of stable isotope fractionation during substrate degradation.Intrinsic microbial degradation is an important process in elimination of contaminants in polluted aquifers, which can be used for the sustainable cleanup of contaminated sites. However, cost-effective remediation strategies such as natural attenuation require a profound knowledge of the microbial degradation processes in the subsurface. Although biodegradation of aromatic hydrocarbons by aerobic and anaerobic bacteria has been investigated in detail in laboratory systems (11,32), assessment at field sites remains difficult. Stable carbon isotope analysis is one approach to quantify microbial activities in situ. For laboratory cultures, isotope fractionation has been shown to occur during degradation of aromatic hydrocarbons, such as toluene (1,19,20), or chlorinated hydrocarbons, such as trichloroethene (3,7,26). In addition, in contaminated field sites, carbon isotope fractionation could be observed and was interpreted to be indicative of microbial degradation in situ (15,24). For toluene as a model compound, it has been demonstrated that isotope fractionation is caused mainly by the first enzyme reaction in the degradation pathway, whereas transport to and into the cell appears not to be relevant for fractionation. The extent of isotope fractionation is considered to be independent of differences in the growth kinetics of the bacteria (20). Isotope fractionation during anaerobic degradation of toluene was on the same order of magnitude for denitrifying, iron (III)-reducing, sulfate-reducing, and fermenting bacteria (1, 19), probably because, in these cases, degradation was initiated by benzylsuccinate synthase. This finding suggests that, in anoxic environments, isotope fractionation could be applied to assess biological degradation quantitatively, as has been worked out recently for several aquifers (23, 25).The objective of this study was to examine whether carbon and hydrogen isotope fractionation could be used to quantify intrinsic biodegradation as well in oxic environments. Previous studies with the aerobic bacterium Pseudomonas putida strain mt-2 showed an extent of isotope fractionation similar to that of anaerobic toluene-degrading strains (19), whereas isotope fractionation during toluene degradation by undefined aerobic microbial communities was not detected (26). Therefore, we started a systematic investigation of the effects of different oxygenase enzymes and stable isotope fractionation.P. putida strain mt-2 (20), Ralstonia pickettii strain PKO1 (J. J. Kukor, Rutgers University, New Brunswick, N.J.), and P. putida strain F1 (A. M. Cook, Konstanz, German...

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supporting

confidence: 91%

Carbon and Hydrogen Stable Isotope Fractionation during Aerobic Bacterial Degradation of Aromatic Hydrocarbons

Morasch

Richnow

Schink

et al. 2002

Appl Environ Microbiol

121

109

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“…While no other study has, to date, reported values for isotopic enrichment factors for anaerobic biodegradation of benzene, carbon (26,53) and hydrogen (26) isotopic enrichment factors have been published for aerobic biodegradation of benzene. For aerobic biodegradation by two bacterial isolates, Hunkeler et al (26) reported a range in carbon enrichment factors (Ϫ1.46 to Ϫ3.53‰) similar to that obtained for anaerobic biodegradation of benzene in this study (Ϫ1.9 to Ϫ3.6‰).…”

Section: Discussionmentioning

confidence: 99%

“…In a study examining hydrogen isotopic fractionation during biodegradation of toluene in pure cultures with various terminal-electronaccepting processes, a small range of hydrogen enrichment factors was reported (40). For aerobic biodegradation of benzene, carbon (26,53) and hydrogen (26) isotopic fractionation has been documented in the literature. However, to date there have been no studies of carbon and hydrogen isotopic fractionation during anaerobic benzene biodegradation.…”

mentioning

confidence: 99%

Carbon and Hydrogen Isotopic Fractionation during Anaerobic Biodegradation of Benzene

Mancini

Ulrich²,

Lacrampe-Couloume

et al. 2003

Appl Environ Microbiol

162

143

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Compound-specific isotope analysis has the potential to distinguish physical from biological attenuation processes in the subsurface. In this study, carbon and hydrogen isotopic fractionation effects during biodegradation of benzene under anaerobic conditions with different terminal-electron-accepting processes are reported for the first time. Different enrichment factors () for carbon (range of ؊1.9 to ؊3.6‰) and hydrogen (range of ؊29 to ؊79‰) fractionation were observed during biodegradation of benzene under nitratereducing, sulfate-reducing, and methanogenic conditions. These differences are not related to differences in initial biomass or in rates of biodegradation. Carbon isotopic enrichment factors for anaerobic benzene biodegradation in this study are comparable to those previously published for aerobic benzene biodegradation. In contrast, hydrogen enrichment factors determined for anaerobic benzene biodegradation are significantly larger than those previously published for benzene biodegradation under aerobic conditions. A fundamental difference in the previously proposed initial step of aerobic versus proposed anaerobic biodegradation pathways may account for these differences in hydrogen isotopic fractionation. Potentially, C-H bond breakage in the initial step of the anaerobic benzene biodegradation pathway may account for the large fractionation observed compared to that in aerobic benzene biodegradation. Despite some differences in reported enrichment factors between cultures with different terminal-electron-accepting processes, carbon and hydrogen isotope analysis has the potential to provide direct evidence of anaerobic biodegradation of benzene in the field.

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“…In groundwater studies, compound-specific isotope analysis (CSIA) has been used increasingly to demonstrate in situ biodegradation of various types of organic contaminants (Kelley et al, 1997;Sturchio et al, 1998;Hunkeler et al, 1999;Sherwood Lollar et al, 2001;Kolhatkar et al, 2002;Mancini et al, 2002;Meckenstock et al, 2002;Song et al, 2002;Kirtland et al, 2003;Richnow et al, 2003a,b;Chu et al, 2004;Griebler et al, 2004;Steinbach et al, 2004;Hunkeler et al, 2005;Morrill et al, 2005). In contrast, only a few studies have investigated the use of CSIA in the unsaturated zone (Stehmeier et al, 1999;Kirtland et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

Carbon isotope fractionation during aerobic biodegradation of n-alkanes and aromatic compounds in unsaturated sand

Bouchard

Hunkeler

Höhener

2008

Organic Geochemistry

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Microcosm experiments were conducted to quantify carbon isotope fractionation during aerobic biodegradation of n-alkanes (from C 3 to C 10 ) and monoaromatic hydrocarbons in unsaturated alluvial sand. In single compound experiments with n-alkanes, the largest enrichment factor was obtained for propane (À10.8 ± 0.7‰). The magnitude of the enrichment factor decreased with increasing number of carbon atoms from propane to n-decane (À0.2 ± 0.1‰). This trend can partly be explained by the decreasing probability that a 13 C is located at the reacting site in the molecule with increasing chain length. After correcting for the presence of non-reacting positions, a chain length dependence of the calculated apparent isotope effect persisted. This observation suggests that transport and binding steps before the actual reaction step become increasingly rate limiting with increasing chain length. For aromatic compounds tested individually, the enrichment factor was the largest (À1.4 ± 0.1‰) for benzene (B), followed by toluene (T) (À0.8 ± 0.1‰) and m-xylene (X) (À0.6 ± 0.1‰). Enrichment factors for BTX were systematically smaller than for n-alkanes with equivalent number of carbons, which is likely related to different biodegradation mechanisms. The study demonstrates that significant carbon isotope fractionation occurs during aerobic biodegradation of n-alkanes and aromatic compounds under unsaturated conditions and that the magnitude of isotope enrichment is linked to molecule size and molecule structure.

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Field and in vitro evidence for in-situ bioremediation using compound-specific 13C/12C ratio monitoring

Cited by 55 publications

References 13 publications

Carbon and Hydrogen Stable Isotope Fractionation during Aerobic Bacterial Degradation of Aromatic Hydrocarbons

Carbon and Hydrogen Stable Isotope Fractionation during Aerobic Bacterial Degradation of Aromatic Hydrocarbons

Carbon and Hydrogen Isotopic Fractionation during Anaerobic Biodegradation of Benzene

Carbon isotope fractionation during aerobic biodegradation of n-alkanes and aromatic compounds in unsaturated sand

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