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. 1993 Nov;59(11):3790–3797. doi: 10.1128/aem.59.11.3790-3797.1993

Dichloromethane as the sole carbon source for an acetogenic mixed culture and isolation of a fermentative, dichloromethane-degrading bacterium.

S A Braus-Stromeyer 1, R Hermann 1, A M Cook 1, T Leisinger 1
PMCID: PMC182533  PMID: 8285685

Abstract

Dichloromethane (DCM) is utilized by the strictly anaerobic, acetogenic mixed culture DM as a sole source of carbon and energy for growth. Growth with DCM was linear, and cell suspensions of the culture degraded DCM with a specific activity of 0.47 mkat/kg of protein. A mass balance of 2 mol of chloride and 0.42 mol of acetate per mol of DCM was observed. The dehalogenation reaction showed similar specific activities under both anaerobic and aerobic conditions. Radioactivity from [14C]DCM in cell suspensions was recovered largely as 14CO2 (58%), [14C]acetate (23%), and [14C]formate (11%), which subsequently disappeared. This suggested that formate is a major intermediate in the pathway from DCM to acetate. Efforts to isolate from culture DM a pure culture capable of anaerobic growth with DCM were unsuccessful, although overall acetogenesis and the partial reactions are thermodynamically favorable. We then isolated bacterial strains DMA, a strictly anaerobic, gram-positive, endospore-forming rod, and DMB, a strictly anaerobic, gram-negative, endospore-forming homoacetogen, from culture DM. Both strain DMB and Methanospirillum hungatei utilized formate as a source of carbon and energy. Coculture of strain DMA with either M. hungatei or strain DMB in solid medium with DCM as the sole added source of carbon and energy was observed. These data support a tentative scheme for the acetogenic fermentation of DCM involving interspecies formate transfer from strain DMA to the acetogenic bacterium DMB or to the methanogen M. hungatei.

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Selected References

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  1. Balch W. E., Fox G. E., Magrum L. J., Woese C. R., Wolfe R. S. Methanogens: reevaluation of a unique biological group. Microbiol Rev. 1979 Jun;43(2):260–296. doi: 10.1128/mr.43.2.260-296.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brunner W., Staub D., Leisinger T. Bacterial degradation of dichloromethane. Appl Environ Microbiol. 1980 Nov;40(5):950–958. doi: 10.1128/aem.40.5.950-958.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Egli C., Tschan T., Scholtz R., Cook A. M., Leisinger T. Transformation of tetrachloromethane to dichloromethane and carbon dioxide by Acetobacterium woodii. Appl Environ Microbiol. 1988 Nov;54(11):2819–2824. doi: 10.1128/aem.54.11.2819-2824.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fahey R. C., Sundquist A. R. Evolution of glutathione metabolism. Adv Enzymol Relat Areas Mol Biol. 1991;64:1–53. doi: 10.1002/9780470123102.ch1. [DOI] [PubMed] [Google Scholar]
  5. Freedman D. L., Gossett J. M. Biodegradation of dichloromethane and its utilization as a growth substrate under methanogenic conditions. Appl Environ Microbiol. 1991 Oct;57(10):2847–2857. doi: 10.1128/aem.57.10.2847-2857.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kennedy S. I., Fewson C. A. Enzymes of the mandelate pathway in Bacterium N.C.I.B. 8250. Biochem J. 1968 Apr;107(4):497–506. doi: 10.1042/bj1070497. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. LaPat-Polasko L. T., McCarty P. L., Zehnder A. J. Secondary substrate utilization of methylene chloride by an isolated strain of Pseudomonas sp. Appl Environ Microbiol. 1984 Apr;47(4):825–830. doi: 10.1128/aem.47.4.825-830.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Müller M., Meister N., Moor H. Freezing in a propane jet and its application in freeze-fracturing. Mikroskopie. 1980 Sep;36(5-6):129–140. [PubMed] [Google Scholar]
  9. Scholtz R., Wackett L. P., Egli C., Cook A. M., Leisinger T. Dichloromethane dehalogenase with improved catalytic activity isolated from a fast-growing dichloromethane-utilizing bacterium. J Bacteriol. 1988 Dec;170(12):5698–5704. doi: 10.1128/jb.170.12.5698-5704.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Schönheit P., Moll J., Thauer R. K. Nickel, cobalt, and molybdenum requirement for growth of Methanobacterium thermoautotrophicum. Arch Microbiol. 1979 Oct;123(1):105–107. doi: 10.1007/BF00403508. [DOI] [PubMed] [Google Scholar]
  11. Stromeyer S. A., Winkelbauer W., Kohler H., Cook A. M., Leisinger T. Dichloromethane utilized by an anaerobic mixed culture: acetogenesis and methanogenesis. Biodegradation. 1991;2(2):129–137. doi: 10.1007/BF00114603. [DOI] [PubMed] [Google Scholar]
  12. Studer D., Michel M., Müller M. High pressure freezing comes of age. Scanning Microsc Suppl. 1989;3:253–269. [PubMed] [Google Scholar]
  13. Thauer R. K., Jungermann K., Decker K. Energy conservation in chemotrophic anaerobic bacteria. Bacteriol Rev. 1977 Mar;41(1):100–180. doi: 10.1128/br.41.1.100-180.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Woese C. R. Bacterial evolution. Microbiol Rev. 1987 Jun;51(2):221–271. doi: 10.1128/mr.51.2.221-271.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]

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