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. 1997 Jan;63(1):296–300. doi: 10.1128/aem.63.1.296-300.1997

Enumeration of Acetogens by a Colorimetric Most-Probable-Number Assay

O T Harriott, A C Frazer
PMCID: PMC1389107  PMID: 16535493

Abstract

Anaerobic O demethylation by acetogenic bacteria often is the first step in the mineralization of methoxylated aromatic compounds in anoxic environments. In this reaction, an ether bond is cleaved and the resulting methyl group is metabolized via the acetyl coenzyme A pathway (acetogenesis). Anaerobic O demethylation was used to assess acetogen populations. Environmental samples were diluted in anaerobic medium containing a methoxylated aromatic substrate (vanillate) and titanium(III), and acetogen titers were estimated by the most-probable-number (MPN) method. Complex formation between Ti(III) and vicinal hydroxyl groups of the aromatic products of anaerobic O demethylation results in the development of a yellow color in the medium, which can be detected by eye and monitored spectrophotometrically. High-performance liquid chromatography analysis of the yellow MPN tubes showed that they contained the product of anaerobic O demethylation of vanillate (protocatechuate). This assay was used to enumerate O-demethylating acetogen populations in environmental samples.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Braun M., Schoberth S., Gottschalk G. Enumeration of bacteria forming acetate from H2 and CO2 in anaerobic habitats. Arch Microbiol. 1979 Mar 12;120(3):201–204. doi: 10.1007/BF00423066. [DOI] [PubMed] [Google Scholar]
  2. COCHRAN W. G. Estimation of bacterial densities by means of the "most probable number". Biometrics. 1950 Jun;6(2):105–116. [PubMed] [Google Scholar]
  3. Doré J., Morvan B., Rieu-Lesme F., Goderel I., Gouet P., Pochart P. Most probable number enumeration of H2-utilizing acetogenic bacteria from the digestive tract of animals and man. FEMS Microbiol Lett. 1995 Jul 15;130(1):7–12. doi: 10.1016/0378-1097(95)00176-6. [DOI] [PubMed] [Google Scholar]
  4. Kuever J., Kulmer J., Jannsen S., Fischer U., Blotevogel K. H. Isolation and characterization of a new spore-forming sulfate-reducing bacterium growing by complete oxidation of catechol. Arch Microbiol. 1993;159(3):282–288. doi: 10.1007/BF00248485. [DOI] [PubMed] [Google Scholar]
  5. Rowe R., Todd R., Waide J. Microtechnique for most-probable-number analysis. Appl Environ Microbiol. 1977 Mar;33(3):675–680. doi: 10.1128/aem.33.3.675-680.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Tasaki M., Kamagata Y., Nakamura K., Mikami E. Utilization of methoxylated benzoates and formation of intermediates by Desulfotomaculum thermobenzoicum in the presence or absence of sulfate. Arch Microbiol. 1992;157(3):209–212. doi: 10.1007/BF00245151. [DOI] [PubMed] [Google Scholar]
  7. Taylor B. F. Aerobic and Anaerobic Catabolism of Vanillic Acid and Some Other Methoxy-Aromatic Compounds by Pseudomonas sp. Strain PN-1. Appl Environ Microbiol. 1983 Dec;46(6):1286–1292. doi: 10.1128/aem.46.6.1286-1292.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Wagner C., Griesshammer A., Drake H. L. Acetogenic capacities and the anaerobic turnover of carbon in a kansas prairie soil. Appl Environ Microbiol. 1996 Feb;62(2):494–500. doi: 10.1128/aem.62.2.494-500.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]

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