Abstract
A series of single-well push–pull tests (SWPPTs) were performed to investigate the efficacy of isobutane (2-methylpropane) as a primary substrate for in situ stimulation of microorganisms able to cometabolically transform common groundwater contaminants, such as chlorinated aliphatic hydrocarbons and 1,4-dioxane (1,4-D). In biostimulation tests, the disappearance of isobutane relative to a nonreactive bromide tracer indicated an isobutane-utilizing microbial community rapidly developed in the aquifer around the test well. SWPPTs were performed as natural drift tests with first-order rates of isobutane consumption ranging from 0.4 to 1.4 day−1. Because groundwater contaminants were not present at the demonstration site, isobutene (2-methylpropene) was used as a nontoxic surrogate to demonstrate cometabolic activity in the subsurface after biostimulation. The transformation of isobutene to isobutene epoxide (2-methyl-1,2-epoxypropane) illustrates the epoxidation process previously shown for common groundwater contaminants after cometabolic transformation by alkane-utilizing bacteria. The rate and extent of isobutene consumption and the formation and transformation of isobutene epoxide were greater in the presence of isobutane, with no evidence of primary substrate inhibition. Modeled concentrations of isobutane-utilizing biomass in microcosms constructed with groundwater collected before and after each SWPPT offered additional evidence that the isobutane-utilizing microbial community was stimulated in the aquifer. Experiments in groundwater microcosms also demonstrated that the isobutane-utilizing bacteria stimulated in the subsurface could cometabolically transform a mixture of co-substrates including isobutene, 1,1-dichloroethene, cis-1,2-dichloroethene, and 1,4-D with the same co-substrate preferences as the bacterium Rhodococcus rhodochrous ATCC strain 21198 after growth on isobutane. This study demonstrated the effectiveness of isobutane as primary substrate for stimulating in situ cometabolic activity and the use of isobutene as surrogate to investigate in situ cometabolic reactions catalyzed by isobutane-stimulated bacteria.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Agency for Toxic Substances and Disease Registry (1996) Toxicological profile for 1,2-dichloroethene. Agency for Toxic Substances and Disease Registry, Atlanta
Agency for Toxic Substances and Disease Registry (2006) Toxicological profile for 1,1,1-trichloroethane. Agency for Toxic Substances and Disease Registry, Atlanta
Agency for Toxic Substances and Disease Registry (2019) Toxicological profile for 1,1-dichloroethene. Agency for Toxic Substances and Disease Registry, Atlanta
Alvarez-Cohen L, Speitel GE (2001) Kinetics of aerobic cometabolism of chlorinated solvents. Biodegradation 12:105–126. https://doi.org/10.1023/A:1012075322466
Anderson JE, McCarty PL (1996) Effect of three chlorinated ethenes on growth rates for a methanotrophic mixed culture. Environ Sci Technol 30:3517–3524. https://doi.org/10.1021/es960187n
Azizian MF, Istok JD, Semprini L (2007) Evaluation of the in-situ aerobic cometabolism of chlorinated ethenes by toluene-utilizing microorganisms using push–pull tests. J Contam Hydrol 90:105–124. https://doi.org/10.1016/j.jconhyd.2006.09.015
Bedient PB, Rifai HS, Newell CJ (2000) Ground water contamination: transport and remediation, 2nd edn. Pearson, London
Bekins BA, Warren E, Godsy EM (1998) A comparison of zero-order, first-order, and Monod biotransformation models. Ground Water 36:261–268. https://doi.org/10.1111/j.1745-6584.1998.tb01091.x
Bennett P, Hyman M, Smith C, El Mugammar H, Chu M-Y, Nickelsen M, Aravena R (2018) Enrichment with carbon-13 and deuterium during monooxygenase-mediated biodegradation of 1,4-dioxane. Environ Sci Technol Lett 5:148–153. https://doi.org/10.1021/acs.estlett.7b00565
Boisson A, de Anna P, Bour O, Le Borgne T, Labasque T, Aquilina L (2013) Reaction chain modeling of denitrification reactions during a push-pull test. J Contam Hydrol 148:1–11. https://doi.org/10.1016/j.jconhyd.2013.02.006
Chang HL, Alvarez-Cohen L (1996) Biodegradation of individual and multiple chlorinated aliphatic hydrocarbons by methane-oxidizing cultures. Appl Env Microbiol 62:3371–3377
Chen W, Faulkner N, Smith C, Fruchte M, Hyman M (2021) Draft genome sequences of four aerobic isobutane-metabolizing bacteria. Microbiol Resour Announc 10:e01381-e1420. https://doi.org/10.1128/MRA.01381-20
Chiang S-YD, Mora R, Diguiseppi WH, Davis G, Sublette K, Gedalanga P, Mahendra S (2012) Characterizing the intrinsic bioremediation potential of 1,4-dioxane and trichloroethene using innovative environmental diagnostic tools. J Environ Monit 14:2317–2326. https://doi.org/10.1039/C2EM30358B
Dalton H, Stirling DI (1982) Co-metabolism. Philos Trans R Soc Lond B Biol Sci 297:481–496. https://doi.org/10.1098/rstb.1982.0056
Deng D, Li F, Wu C, Li M (2018) Synchronic biotransformation of 1,4-dioxane and 1,1-dichloroethylene by a gram-negative propanotroph Azoarcus sp. DD4. Sci. Technol. Lett, Environ. https://doi.org/10.1021/acs.estlett.8b00312
Dolan ME, McCarty PL (1995) Methanotrophic chloroethene transformation capacities and 1,1-dichloroethene transformation product toxicity. Environ Sci Technol 29:2741–2747. https://doi.org/10.1021/es00011a007
Doughty DM, Sayavedra-Soto LA, Arp DJ, Bottomley PJ (2005) Effects of dichloroethene isomers on the induction and activity of butane monooxygenase in the alkane-oxidizing bacterium “Pseudomonas butanovora”. Appl Environ Microbiol 71:6054–6059. https://doi.org/10.1128/AEM.71.10.6054-6059.2005
Draper WM, Dhoot JS, Remoy JW, Perera SK (2000) Trace-level determination of 1,4-dioxane in water by isotopic dilution GC and GC-MS. Analyst 125:1403–1408
Dupin HJ, McCarty PL (2000) Impact of colony morphologies and disinfection on biological clogging in porous media. Environ Sci Technol 34:1513–1520. https://doi.org/10.1021/es990452f
Ely RL, Williamson KJ, Hyman MR, Arp DJ (1997) Cometabolism of chlorinated solvents by nitrifying bacteria: kinetics, substrate interactions, toxicity effects, and bacterial response. Biotechnol Bioeng 54:520–534. https://doi.org/10.1002/(SICI)1097-0290(19970620)54:6%3c520::AID-BIT3%3e3.0.CO;2-L
Ennis E, Reed R, Dolan M, Semprini L, Istok J, Field J (2005) Reductive dechlorination of the vinyl chloride surrogate chlorofluoroethene in TCE-contaminated groundwater. Environ Sci Technol 39:6777–6785. https://doi.org/10.1021/es048640f
Frascari D, Zanaroli G, Danko AS (2015) In situ aerobic cometabolism of chlorinated solvents: a review. J Hazard Mater 283:382–399. https://doi.org/10.1016/j.jhazmat.2014.09.041
Hageman KJ, Istok JD, Field JA, Buscheck TE, Semprini L (2001) In situ anaerobic transformation of trichlorofluoroethene in trichloroethene-contaminated groundwater. Environ Sci Technol 35:1729–1735
Hageman KJ, Field JA, Istok JD, Schroth MH (2003) “Forced mass balance” technique for estimating in situ transformation rates of sorbing solutes in groundwater. Environ Sci Technol 37:3920–3925. https://doi.org/10.1021/es0342042
Haggerty R, Schroth MH, Istok JD (1998) Simplified method of “push-pull” test data analysis for determining in situ reaction rate coefficients. Groundwater 36:314–324. https://doi.org/10.1111/j.1745-6584.1998.tb01097.x
Halsey KH, Doughty DM, Sayavedra-Soto LA, Bottomley PJ, Arp DJ (2007) Evidence for modified mechanisms of chloroethene oxidation in Pseudomonas butanovora mutants containing single amino acid substitutions in the hydroxylase alpha-subunit of butane monooxygenase. J Bacteriol 189:5068–5074. https://doi.org/10.1128/JB.00189-07
Hatzinger PB, Banerjee R, Rezes R, Streger SH, McClay K, Schaefer CE (2017) Potential for cometabolic biodegradation of 1,4-dioxane in aquifers with methane or ethane as primary substrates. Biodegradation 28:453–468. https://doi.org/10.1007/s10532-017-9808-7
Hazen TC (2010) Cometabolic bioremediation. In: Timmis KN (ed) Handbook of hydrocarbon and lipid microbiology. Springer, Berlin, pp 2505–2514. https://doi.org/10.1007/978-3-540-77587-4_185
Hazen TC, Jiménez L, de Victoria GL, Fliermans CB (1991) Comparison of bacteria from deep subsurface sediment and adjacent groundwater. Microb Ecol 22:293–304
Hopkins GD, McCarty PL (1995) Field evaluation of in situ aerobic cometabolism of trichloroethylene and three dichloroethylene isomers using phenol and toluene as the primary substrates. Environ Sci Technol 29:1628–1637. https://doi.org/10.1021/es00006a029
Huling SG, Bledsoe BE (1990) Enhanced bioremediation utilizng hydrogen peroxide as a supplemental source of oxygen: a laboratory and field study (No. EPA/600/2-90/006). Environmental Protection Agency, Robert S. Kerr Environmental Research Laboratory, Ada, OK
Huntscha S, Rodriguez Velosa DM, Schroth MH, Hollender J (2013) Degradation of polar organic micropollutants during riverbank filtration: complementary results from spatiotemporal sampling and push–pull tests. Environ Sci Technol 47:11512–11521. https://doi.org/10.1021/es401802z
Hyman MR, Wood PM (1985) Suicidal inactivation and labelling of ammonia mono-oxygenase by acetylene. Biochem J 227:719–725
Hyman M, Taylor C, O’Reilly KT (2000) Cometabolic degradation of MTBE by iso-alkane-utilizing bacteria from gasoline-impacted soils, in: Bioremediation and Phytoremediation of Chlorinated and Recalcitrant Compounds. Presented at the international conference on remediation of chlorinated and recalcitrant compounds, Battelle Press, Monterey, California, pp 149–155
Inoue D, Tsunoda T, Sawada K, Yamamoto N, Saito Y, Sei K, Ike M (2016) 1,4-Dioxane degradation potential of members of the genera Pseudonocardia and Rhodococcus. Biodegradation. https://doi.org/10.1007/s10532-016-9772-7
International Programme on Chemical Safety (1998) ICSC 0901—ISOBUTANE [WWW Document]. INCHEM. http://www.inchem.org/documents/icsc/icsc/eics0901.htm. Accessed 7 June 2021
International Programme on Chemical Safety (2000) ICSC 1027 - ISOBUTENE [WWW Document]. INCHEM. URL http://www.inchem.org/documents/icsc/icsc/eics1027.htm. Accessed 23 June 2021
International Programme on Chemical Safety (2003) ICSC 0319—PROPANE [WWW Document]. https://inchem.org/documents/icsc/icsc/eics0319.htm. Accessed 10 June 2021
Istok JD (2013) Push–pull tests for site characterization. Lecture notes in Earth system sciences. Springer, Berlin. https://doi.org/10.1007/978-3-642-13920-8
Istok JD, Humphrey MD, Schroth MH, Hyman MR, O’Reilly KT (1997) Single-well, “push–pull” test for in situ determination of microbial activities. Groundwater 35:619–631. https://doi.org/10.1111/j.1745-6584.1997.tb00127.x
Jesus J, Frascari D, Pozdniakova T, Danko AS (2016) Kinetics of aerobic cometabolic biodegradation of chlorinated and brominated aliphatic hydrocarbons: a review. J Hazard Mater 309:37–52. https://doi.org/10.1016/j.jhazmat.2016.01.065
Johnson NW, Gedalanga PB, Zhao L, Gu B, Mahendra S (2020) Cometabolic biotransformation of 1,4-dioxane in mixtures with hexavalent chromium using attached and planktonic bacteria. Sci Tot Environ 706:135734. https://doi.org/10.1016/j.scitotenv.2019.135734
Kim Y, Semprini L (2005) Cometabolic transformation of cis-1,2-dichloroethylene and cis-1,2-dichloroethylene epoxide by a butane-grown mixed culture. Water Sci Technol J Int Assoc Water Pollut Res 52:125–131
Kim Y, Arp DJ, Semprini L (2002) A combined method for determining inhibition type, kinetic parameters, and inhibition coefficients for aerobic cometabolism of 1,1,1-trichloroethane by a butane-grown mixed culture. Biotechnol Bioeng 77:564–576
Kim Y, Istok JD, Semprini L (2004) Push–pull tests for assessing in situ aerobic cometabolism. Ground Water 42:329–337
Kim Y, Azizian MF, Istok J, Semprini L (2005) Field push-pull test protocol for aerobic cometabolism of chlorinated aliphatic hydrocarbons. Environmental Security Technology Certification Program, Arlington
Kim Y, Istok JD, Semprini L (2006) Push–pull tests evaluating in situ aerobic cometabolism of ethylene, propylene, and cis-1,2-dichloroethylene. J Contam Hydrol 82:165–181. https://doi.org/10.1016/j.jconhyd.2005.10.003
Kim Y, Istok JD, Semprini L (2008) Single-well, gas-sparging tests for evaluating the in situ aerobic cometabolism of cis-1,2-dichloroethene and trichloroethene. Chemosphere 71:1654–1664. https://doi.org/10.1016/j.chemosphere.2008.01.021
Kottegoda S, Waligora E, Hyman M (2015) Metabolism of 2-methylpropene (isobutylene) by the aerobic bacterium Mycobacterium sp. strain ELW1. Appl Environ Microbiol 81:1966–1976. https://doi.org/10.1128/AEM.03103-14
Krippaehne KJ (2018) Cometabolism of 1,4-dioxane and chlorinated aliphatic hydrocarbons by pure cultures of Rhodococcus rhodochrous 21198 and Mycobacterium ELW1 and in groundwater microcosms fed isobutane and isobutene as growth substrates. Oregon State University, Corvallis
Laurance JC (2019) Bioaugmentation of pure and enriched cultures fed multiple primary substrates to aerobically cometabolize a mixture of chlorinated solvents and 1,4-dioxane in microcosms containing groundwater from a department of defense site. Oregon State University, Corvallis
Lehman RM (2007) Understanding of aquifer microbiology is tightly linked to sampling approaches. Geomicrobiol J 24:331–341. https://doi.org/10.1080/01490450701456941
Li F, Deng D, Zeng L, Abrams S, Li M (2021) Sequential anaerobic and aerobic bioaugmentation for commingled groundwater contamination of trichloroethene and 1,4-dioxane. Sci Tot Environ 774:145118. https://doi.org/10.1016/j.scitotenv.2021.145118
Mahendra S, Alvarez-Cohen L (2006) Kinetics of 1,4-dioxane biodegradation by monooxygenase-expressing bacteria. Environ Sci Technol 40:5435–5442. https://doi.org/10.1021/es060714v
Michalsen MM, Weiss R, King A, Gent D, Medina VF, Istok JD (2013) Push-pull tests for estimating RDX and TNT degradation rates in groundwater. Groundw Monit Remediat 33:61–68. https://doi.org/10.1111/gwmr.12016
Mohr TKG, Stickney JA, DiGuiseppi WH (2010) Environmental investigation and remediation: 1,4-dioxane and other solvent stabilizers. CRC Press, Boca Raton
Moreno R, Rojo F (2019) Enzymes for aerobic degradation of alkanes in bacteria. In: Aerobic utilization of hydrocarbons, oils, and lipids, handbook of hydrocarbon and lipid microbiology. Springer, Berlin
Mujica-Alarcon JF, Thornton SF, Rolfe SA (2021) Long-term dynamic changes in attached and planktonic microbial communities in a contaminated aquifer. Environ Pollut 277:116765. https://doi.org/10.1016/j.envpol.2021.116765
Murnane RA, Chen W, Hyman M, Semprini L (2021) Long-term cometabolic transformation of 1,1,1-trichloroethane and 1,4-dioxane by Rhodococcus rhodochrous ATCC 21198 grown on alcohols slowly produced by orthosilicates. J Contam Hydrol 240:103796. https://doi.org/10.1016/j.jconhyd.2021.103796
Nicholls HCG, Rolfe SA, Mallinson HEH, Hjort M, Spence MJ, Bonte M, Thornton SF (2021) Distribution of ETBE-degrading microorganisms and functional capability in groundwater, and implications for characterising aquifer ETBE biodegradation potential. Environ Sci Pollut Res. https://doi.org/10.1007/s11356-021-15606-7
Phanikumar MS, McGuire JT (2010) A multi-species reactive transport model to estimate biogeochemical rates based on single-well push–pull test data. Comput Geosci 36:997–1004. https://doi.org/10.1016/j.cageo.2010.04.001
Prince RC, Parkerton TF, Lee C (2007) The primary aerobic biodegradation of gasoline hydrocarbons. Environ Sci Technol 41:3316–3321. https://doi.org/10.1021/es062884d
Reusser DE, Istok JD, Beller HR, Field JA (2002) In situ transformation of deuterated toluene and xylene to benzylsuccinic acid analogues in BTEX-contaminated aquifers. Environ Sci Technol 36:4127–4134. https://doi.org/10.1021/es0257366
Rolston HM, Hyman MR, Semprini L (2019) Aerobic cometabolism of 1,4-dioxane by isobutane-utilizing microorganisms including Rhodococcus rhodochrous strain 21198 in aquifer microcosms: Experimental and modeling study. Sci Tot Environ. https://doi.org/10.1016/j.scitotenv.2019.133688
Sander R (2015) Compilation of Henry’s law constants (version 4.0) for water as solvent. Atmos Chem Phys 15:4399–4981. https://doi.org/10.5194/acp-15-4399-2015
Schrlau JE, Kramer AL, Chlebowski A, Truong L, Tanguay RL, Simonich SLM, Semprini L (2017) Formation of developmentally toxic phenanthrene metabolite mixtures by Mycobacterium sp. ELW1. Environ Sci Technol 51:8569–8578. https://doi.org/10.1021/acs.est.7b01377
Schroth M, Istok J, Hyman M, O’Reilly K, Battelle Memorial Institute (1997) Field-scale measurements of in situ microbial metabolic activities. In: In situ and on-site bioremediation, vol 2. Battelle Press, New Orleans, pp 387–392
Semprini L (1997) Strategies for the aerobic co-metabolism of chlorinated solvents. Curr Opin Biotechnol 8:296–308
Semprini L, Dolan ME, Mathias MA, Hopkins GD, McCarty PL (2007) Laboratory, field, and modeling studies of bioaugmentation of butane-utilizing microorganisms for the in situ cometabolic treatment of 1,1-dichloroethene, 1,1-dichloroethane, and 1,1,1-trichloroethane. Adv Water Resour 30:1528–1546. https://doi.org/10.1016/j.advwatres.2006.05.017
Shennan JL (2006) Utilisation of C2–C4 gaseous hydrocarbons and isoprene by microorganisms. J Chem Technol Biotechnol 81:237–256. https://doi.org/10.1002/jctb.1388
Shi W, Wang Q, Zhan H (2020) New simplified models of single-well push-pull tests with mixing effect. Water Resour Res. https://doi.org/10.1029/2019WR026802
Shields-Menard SA, Brown SD, Klingeman DM, Indest K, Hancock D, Wewalwela JJ, French WT, Donaldson JR (2014) Draft genome sequence of Rhodococcus rhodochrous strain ATCC 21198. Genome Announc. https://doi.org/10.1128/genomeA.00054-14
Smith HJ, Zelaya AJ, De León KB, Chakraborty R, Elias DA, Hazen TC, Arkin AP, Cunningham AB, Fields MW (2018) Impact of hydrologic boundaries on microbial planktonic and biofilm communities in shallow terrestrial subsurface environments. FEMS Microbiol Ecol. https://doi.org/10.1093/femsec/fiy191
Tovanabootr A, Semprini L (1998) Comparison of long-term TCE transformation ability of methane and propane-utilizing microorganisms stimulated from the McClellan AFB subsurface. Bioremediat J 2:105–124
Weijers CA, de Haan A, de Bont JA (1988) Microbial production and metabolism of epoxides. Microbiol Sci 5:156–159
Woods NR, Murrell JC (1990) Epoxidation of gaseous alkenes by a Rhodococcus sp. Biotechnol Lett 12:409–414. https://doi.org/10.1007/BF01024394
Wu W-M, Watson DB, Luo J, Carley J, Mehlhorn T, Kitanidis PK, Jardine PM, Criddle CS (2013) Surge block method for controlling well clogging and sampling sediment during bioremediation. Water Res 47:6566–6573. https://doi.org/10.1016/j.watres.2013.08.033
Xiong Y, Mason OU, Lowe A, Zhang Z, Zhou C, Chen G, Villalonga MJ, Tang Y (2020) Investigating promising substrates for promoting 1,4-dioxane biodegradation: effects of ethane and tetrahydrofuran on microbial consortia. Biodegradation 31:171–182. https://doi.org/10.1007/s10532-020-09901-2
Zhang S, Gedalanga PB, Mahendra S (2016) Biodegradation kinetics of 1,4-dioxane in chlorinated solvent mixtures. Environ Sci Technol 50:9599–9607. https://doi.org/10.1021/acs.est.6b02797
Zhang S, Gedalanga PB, Mahendra S (2017) Advances in bioremediation of 1,4-dioxane-contaminated waters. J Environ Manage 204:765–774. https://doi.org/10.1016/j.jenvman.2017.05.033
Acknowledgements
The authors gratefully acknowledge Dr. Mohammad Azizian for sharing his expertise in the execution of push-pull tests, Jon Laurance for his assistance with data collection, and Justin Fleming and other staff at the Oregon State University Motor Pool for providing access to the test well.
Funding
This study was funded by the Department of Defense Strategic Environmental Research Development Program (Grant ER-2303), the National Defense Science and Engineering Graduate Fellowship Program, and the DeVaan Fellowship for Clean and Sustainable Water Technology.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no competing or financial interests.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Rolston, H., Hyman, M. & Semprini, L. Single-well push–pull tests evaluating isobutane as a primary substrate for promoting in situ cometabolic biotransformation reactions. Biodegradation 33, 349–371 (2022). https://doi.org/10.1007/s10532-022-09987-w
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10532-022-09987-w