Anaerobic methane oxidation coupled to chromate reduction in a methane-based membrane biofilm batch reactor

  • Qiu-Yi Dong
  • Zhen Wang
  • Ling-Dong Shi
  • Chun-Yu LaiEmail author
  • He-Ping ZhaoEmail author
Research Article


Chromate can be reduced by methanotrophs in a membrane biofilm reactor (MBfR). In this study, we cultivated a Cr(VI)-reducing biofilm in a methane (CH4)-based membrane biofilm batch reactor (MBBR) under anaerobic conditions. The Cr(VI) reduction rate increased to 0.28 mg/L day when the chromate concentration was ≤ 2.2 mg/L but declined sharply to 0.01 mg/L day when the Cr(VI) concentration increased to 6 mg/L. Isotope tracing experiments showed that part of the 13C-labeled CH4 was transformed to 13CO2, suggesting that the biofilm may reduce Cr(VI) by anaerobic methane oxidation (AnMO). Microbial community analysis showed that a methanogen, i.e., Methanobacterium, dominated in the biofilm, suggesting that this genus is probably capable of carrying out AnMO. The abundance of Methylomonas, an aerobic methanotroph, decreased significantly, while Meiothermus, a potential chromate-reducing bacterium, was enriched in the biofilm. Overall, the results showed that the anaerobic environment inhibited the activity of aerobic methanotrophs while promoting AnMO bacterial enrichment, and high Cr(VI) loading reduced Cr(VI) flux by inhibiting the methane oxidation process.


Chromate Methane AnMO Remediation 


Funding information

The authors greatly thank the National Natural Science Foundation of China (grant nos. 21577123, 51878596) and Natural Science Funds for Distinguished Young Scholar of Zhejiang Province (LR17B070001) for their financial support.

Supplementary material

11356_2019_5709_MOESM1_ESM.docx (198 kb)
ESM 1 (DOCX 197 kb)


  1. Ackerley DF, Gonzalez CF, Keyhan M, Blake R II, Matin A (2010) Mechanism of chromate reduction by the Escherichia coli protein, NfsA, and the role of different chromate reductases in minimizing oxidative stress during chromate reduction. Environ Microbiol 6:851–860CrossRefGoogle Scholar
  2. Barnhart J (1997) Occurrences, uses, and properties of chromium. Regul Toxicol Pharmacol 26:3–7CrossRefGoogle Scholar
  3. Cakir FY, Stenstrom MK (2005) Greenhouse gas production: a comparison between aerobic and anaerobic wastewater treatment technology. Water Res 39(17):4197–4203Google Scholar
  4. Diederik JO, Lizelle AP, Esta VH (2008) A novel chromate reductase from Thermus scotoductus SA-01 related to old yellow enzyme. J Bacteriol 190:3076–3082CrossRefGoogle Scholar
  5. Dragun J (1988) The soil chemistry of hazardous materials. Hazardous Materials Control Research Institute, Silver SpringGoogle Scholar
  6. Ettwig KF, Butler MK, Paslier DL, Pelletier E, Mangenot S, Kuypers MMM, Schreiber F, Dutilh BE, Zedelius J, De Beer D (2010) Nitrite-driven anaerobic methane oxidation by oxygenic bacteria. Nature 464:543–548CrossRefGoogle Scholar
  7. Garcia EA, Gomis DB (1997) Speciation analysis of chromium using crypt and ethers. Analyst 122:899–902CrossRefGoogle Scholar
  8. Haroon MF, Hu S, Shi Y, Imelfort M, Keller J, Hugenholtz P, Yuan Z, Tyson GW (2013) Anaerobic oxidation of methane coupled to nitrate reduction in a novel archaeal lineage. Nature 500:567–570CrossRefGoogle Scholar
  9. Hu BL, Shen LD, Lian X, Zhu Q, Liu S, Huang Q, He ZF, Geng S, Cheng DQ, Lou LP, Xu XY, Zheng P, He YF (2014) Evidence for nitrite-dependent anaerobic methane oxidation as a previously overlooked microbial methane sink in wetlands. Proc Natl Acad Sci U S A 111:4495–4500CrossRefGoogle Scholar
  10. Kantar C, Cetin Z, Demiray H (2008) In situ stabilization of chromium(VI) in polluted soils using organic ligands: the role of galacturonic, glucuronic and alginic acids. J Hazard Mater 159:287–293CrossRefGoogle Scholar
  11. Kathiravan MN, Karthick R, Muthukumar K (2011) Ex situ bioremediation of Cr(VI) contaminated soil by Bacillus sp.: batch and continuous studies. Chem Eng J 169:107–115CrossRefGoogle Scholar
  12. Knittel K, Losekann T, Boetius A, Kort R, Amann R (2005) Diversity and distribution of methanotrophic archaea at cold seeps. Appl Environ Microbiol 71:467–479CrossRefGoogle Scholar
  13. Lai CY, Yang X, Tang YN, Rittmann BE, Zhao HP (2014) Nitrate shaped the selenate-reducing microbial community in a hydrogen-based biofilm reactor. Environ Sci Technol 48:3395–3402CrossRefGoogle Scholar
  14. Lai CY, Wen LL, Zhang Y, Luo SS, Wang QY, Luo YH, Chen R, Yang X, Rittmann BE, Zhao HP (2016a) Autotrophic antimonate bio-reduction using hydrogen as the electron donor. Water Res 88:467–474CrossRefGoogle Scholar
  15. Lai CY, Zhong L, Zhang Y, Chen JX, Wen LL, Shi LD, Sun YP, Ma F, Rittmann BE, Zhou C, Tang YN, Zheng P, Zhao HP (2016b) Bioreduction of chromate in a methane-based membrane biofilm reactor. Environ Sci Technol 50:5832–5839CrossRefGoogle Scholar
  16. Lai CY, Dong QY, Chen JX, Zhu QS, Yang X, Chen WD, Zhao HP, Zhu L (2018a) Role of extracellular polymeric substances in a methane based membrane biofilm reactor reducing vanadate. Environ Sci Technol 52:10680–10688CrossRefGoogle Scholar
  17. Lai CY, Dong QY, Rittmann BE, Zhao HP (2018b) Bioreduction of antimonate by anaerobic methane oxidation in a membrane biofilm batch reactor. Environ Sci Technol 52:8693–8700CrossRefGoogle Scholar
  18. Lai CY, Lv PL, Dong QY, Yeo SL, Rittmann B, Zhao HP (2018c) Bromate and nitrate bioreduction coupled with poly-β-hydroxybutyrate production in a methane-based membrane biofilm reactor. Environ Sci Technol 52:7024–7031CrossRefGoogle Scholar
  19. Lovley DR, Coates JD (1997) Bioremediation of metal contamination. Curr Opin Biotechnol 8:285–289CrossRefGoogle Scholar
  20. Lu YZ, Fu L, Ding J, Ding ZW, Li N, Zeng RJ (2016) Cr(VI) reduction coupled with anaerobic oxidation of methane in a laboratory reactor. Water Res 102:445–452CrossRefGoogle Scholar
  21. Luo J, Chen H, Hu S, Cai C, Yuan ZG, Guo JH (2018) Microbial selenate reduction driven by a denitrifying anaerobic methane oxidation biofilm. Environ Sci Technol 52:4006–4012CrossRefGoogle Scholar
  22. Lv PL, Zhong L, Dong QY, Yang SL, Shen WW, Zhu QS, Lai CY, Luo AC, Tang Y, Zhao HP (2018) The effect of electron competition on chromate reduction using methane as electron donor. Environ Sci Pollut Res 25(7):6609–6618CrossRefGoogle Scholar
  23. Lv PL, Shi LD, Wang Z, Rittmann BE, Zhao HP (2019) Methane oxidation coupled to perchlorate reduction in a membrane biofilm batch reactor. Sci Total Environ 667:9–15CrossRefGoogle Scholar
  24. Maeda H, Fujimoto C, Haruki Y, Maeda T, Kokeguchi S, Petelin M, Arai H, Tanimoto I, Nishimura F and Takashiba S, (2003) Quantitative real-time PCR using TaqMan and SYBR Green for Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, tetQ gene and total bacteria. FEMS Immunology & Medical Microbiology, 39(1):81–86Google Scholar
  25. Martin KJ, Nerenberg R (2012) The membrane biofilm reactor (MBfR) for water and wastewater treatment: principles, applications, and recent developments. Bioresour Technol 122:83–94CrossRefGoogle Scholar
  26. Meyer KJ, Swaim P D, Bellamy WD, Rittmann BE, Tang YN, Scott R (2010) Biological and ion exchange nitrate removal: performance and sustainability evaluation, Final Project Report, Water Research Foundation: Denver, COGoogle Scholar
  27. Modin O, Fukushi K, Yamamoto K (2007) Denitrification with methane as external carbon source. Water Res 41:2726–2738CrossRefGoogle Scholar
  28. Moran JJ, House CH, Freeman KH, Ferry JG (2014) Trace methane oxidation studied in several euryarchaeota under diverse conditions. Archaea 5:303Google Scholar
  29. Orphan VJ, House CH, Hinrichs KU, Mckeegan KD, Delong EF (2001) Methane-consuming archaea revealed by directly coupled isotopic and phylogenetic analysis. Science 293:484–487CrossRefGoogle Scholar
  30. Rittmann, McCarty (2002) Environmental biotechnology: principles and applications. McGraw-HillGoogle Scholar
  31. Sahinkaya E, Kilic A, Calimlioglu B, Toker Y (2013) Simultaneous bioreduction of nitrate and chromate using sulfur-based mixotrophic denitrification process. J Hazard Mater 262:234–239CrossRefGoogle Scholar
  32. Smith WA, Apel WA, Petersen JN, Peyton BM (2002) Effect of carbon and energy source on bacterial chromate reduction. Bioremediat J 6:205–215CrossRefGoogle Scholar
  33. Sun Y, Wolcott RD, Dowd SE (2011) Tag-encoded FLX amplicon pyrosequencing for the elucidation of microbial and functional gene diversity in any environment. In: Kwon Y, Ricke S (eds) High-throughput next generation sequencing. Methods in molecular biology (Methods and Protocols) 733. Humana Press, TotowaGoogle Scholar
  34. Viamajala S, Peyton BM, Sani RK, Apel WA, Petersen JN (2008) Toxic effects of chromium(VI) on anaerobic and aerobic growth of Shewanella oneidensis MR-1. Biotechnol Prog 20:87–95CrossRefGoogle Scholar
  35. Wang DB, Wang YL, Liu YW, Ngo HH, Lian Y, Zhao JW, Chen F, Yang Q, Zeng GM, Li XM (2017a) Is denitrifying anaerobic methane oxidation-centered technologies a solution for the sustainable operation of wastewater treatment Plants? Bioresour Technol 234:456–465CrossRefGoogle Scholar
  36. Wang GY, Zhang BG, Li S, Yang M, Yin CC (2017b) Simultaneous microbial reduction of vanadium (V) and chromium (VI) by Shewanella loihica PV-4. Bioresour Technol 227:353–358CrossRefGoogle Scholar
  37. Wang S, Zhang BG, Diao MH, Shi JX, Jiang YF, Cheng YT, Liu H (2018) Enhancement of synchronous bio-reductions of vanadium (V) and chromium (VI) by mixed anaerobic culture. Environ Pollut 242:249–256CrossRefGoogle Scholar
  38. World Health Organization (2011) Guidelines for drinking-water quality, 4th edn. WHO Press, GenevaGoogle Scholar
  39. Zahoor A, Rehman A (2009) Isolation of Cr(VI) reducing bacteria from industrial effluents and their potential use in bioremediation of chromium containing wastewater. J Environ Sci China 21:814–820CrossRefGoogle Scholar
  40. Zehnder AJB, Brock TD (1979) Methane formation and methane oxidation by methanogenic bacteria. J Bacteriol 137:420–432Google Scholar
  41. Zhang BG, Feng CP, Ni JR, Zhang J, Huang WL (2013) Simultaneous reduction of vanadium (V) and chromium (VI) with enhanced energy recovery based on microbial fuel cell technology. J Power Sources 204:34–39CrossRefGoogle Scholar
  42. Zhong L, Lai CY, Shi LD, Wang KD, Dai YJ, Liu YW, Ma F, Rittmann BE, Zheng P, Zhao HP (2017) Nitrate effects on chromate reduction in a methane-based biofilm. Water Res 115:130–137CrossRefGoogle Scholar
  43. Zhou Y, Guo H, Lu H, Mao R, Zheng H, Wang J (2015) Analytical methods and application of stable isotopes in dissolved organic carbon and inorganic carbon in groundwater. Rapid Commun Mass Spectrom 29:1827–1835CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.MOE Key Lab of Environmental Remediation and Ecosystem Health, College of Environmental and Resource ScienceZhejiang UniversityHangzhouChina
  2. 2.Zhejiang Prov Key Lab Water Pollut Control & EnviZhejiang UniversityHangzhouChina
  3. 3.Advanced Water Management CentreThe University of QueenslandSt. LuciaAustralia

Personalised recommendations