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Applied and Environmental Microbiology, October 2008, p. 6187-6193, Vol. 74, No. 20
0099-2240/08/$08.00+0     doi:10.1128/AEM.01142-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Biodegradation of 2-Ethylhexyl Nitrate by Mycobacterium austroafricanum IFP 2173{triangledown}

Elodie Nicolau,1 Lucien Kerhoas,2 Martine Lettere,2 Yves Jouanneau,3 and Rémy Marchal1*

IFP, Département de Biotechnologie et Chimie de la Biomasse, 1-4 avenue du Bois Préau, 92852 Rueil-Malmaison Cedex, France,1 INRA, Unité de Phytopharmacie et Médiateurs Chimiques, Route de St-Cyr, 78026 Versailles Cedex, France,2 CEA, DSV, iRTSV, Laboratoire de Chimie et Biologie des Métaux, 17 rue des Martyrs, F-38054 Grenoble, CNRS, UMR5249, F-38054 Grenoble, and Université Joseph Fourrier, F-38000 Grenoble, France3

Received 21 May 2008/ Accepted 12 August 2008

2-Ethyhexyl nitrate (2-EHN) is a major additive of fuel that is used to increase the cetane number of diesel. Because of its wide use and possible accidental release, 2-EHN is a potential pollutant of the environment. In this study, Mycobacterium austroafricanum IFP 2173 was selected from among several strains as the best 2-EHN degrader. The 2-EHN biodegradation rate was increased in biphasic cultures where the hydrocarbon was dissolved in an inert non-aqueous-phase liquid, suggesting that the transfer of the hydrophobic substrate to the cells was a growth-limiting factor. Carbon balance calculation, as well as organic-carbon measurement, indicated a release of metabolites in the culture medium. Further analysis by gas chromatography revealed that a single metabolite accumulated during growth. This metabolite had a molecular mass of 114 Da as determined by gas chromatography/mass spectrometry and was provisionally identified as 4-ethyldihydrofuran-2(3H)-one by liquid chromatography-tandem mass spectrometry analysis. Identification was confirmed by analysis of the chemically synthesized lactone. Based on these results, a plausible catabolic pathway is proposed whereby 2-EHN is converted to 4-ethyldihydrofuran-2(3H)-one, which cannot be metabolized further by strain IFP 2173. This putative pathway provides an explanation for the low energetic efficiency of 2-EHN degradation and its poor biodegradability.

* Corresponding author. Mailing address: IFP, Département de Biotechnologie, 1-4 avenue Bois-Préau, 92852 Rueil-Malmaison Cedex, France. Phone: 33 (0)1 47 52 69 24. Fax: 33 (0)1 47 52 70 01. E-mail: remy.marchal{at}ifp.fr

{triangledown} Published ahead of print on 22 August 2008.

Applied and Environmental Microbiology, October 2008, p. 6187-6193, Vol. 74, No. 20
0099-2240/08/$08.00+0     doi:10.1128/AEM.01142-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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