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Applied and Environmental Microbiology, September 2005, p. 5171-5176, Vol. 71, No. 9
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.9.5171-5176.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Survey of Extreme Solvent Tolerance in Gram-Positive Cocci: Membrane Fatty Acid Changes in Staphylococcus haemolyticus Grown in Toluene

Lindsey E. Nielsen,1,{dagger} Dana R. Kadavy,1,{ddagger} Soumitra Rajagopal,1,§ Rhae Drijber,2 and Kenneth W. Nickerson1*

School of Biological Sciences, University of Nebraska, Lincoln, Nebraska 68588-0666,1 Department of Agronomy and Horticulture, University of Nebraska, Lincoln, Nebraska 68583-09152

Received 4 December 2004/ Accepted 23 March 2005

We exploited the unique ecological niche of oil fly larval guts to isolate a strain of Staphylococcus haemolyticus which may be the most solvent-tolerant gram-positive bacterium yet described. This organism is able to tolerate 100% toluene, benzene, and p-xylene on plate overlays and saturating levels of these solvents in monophasic liquid cultures. A comparison of membrane fatty acids by gas chromatography after growth in liquid media with and without toluene showed that in cells continuously exposed to solvent the proportion of anteiso fatty acids increased from 25.8 to 33.7% while the proportion of 20:0 straight-chain fatty acids decreased from 19.3 to 10.1%. No changes in the membrane phospholipid composition were noted. Thus, S. haemolyticus alters its membrane fluidity via fatty acid composition to become more fluid when it is exposed to solvent. This response is opposite that commonly found in gram-negative bacteria, which change their fatty acids so that the cytoplasmic membrane is less fluid. Extreme solvent tolerance in S. haemolyticus is not accompanied by abnormal resistance to anionic or cationic detergents. Finally, six strains of Staphylococcus aureus and five strains of Staphylococcus epidermidis, which were not obtained by solvent selection, also exhibited exceptional solvent tolerance.

* Corresponding author. Mailing address: School of Biological Sciences, University of Nebraska, Lincoln, NE 68588-0666. Phone: (402) 472-2253. Fax: (402) 472-8722. E-mail: knickerson1{at}unl.edu.

{dagger} Present address: Interdepartmental Microbiology Program, Iowa State University, Ames, IA 50011.

{ddagger} Present address: Signature Science Inc., Austin, TX 78759.

§ Present address: Department of Molecular Biology & Microbiology, Tufts University, Boston, MA 02111.

Applied and Environmental Microbiology, September 2005, p. 5171-5176, Vol. 71, No. 9
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.9.5171-5176.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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