Measurement of Hydrocarbon Transport in Bacteria
Hydrocarbon uptake by bacteria has not been extensively studied, and strong evidence for active transport of hydrocarbons is lacking. The volatile nature of hydrocarbons, their hydrophobicity, and their relatively low aqueous solubilities can complicate t
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Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4214
2 2.1 2.1.1 2.1.2 2.1.3 2.1.4 2.1.5 2.1.6
Experimental Approach . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4215 Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4215 Preparation of Transport Assay Mixture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4215 Growth and Preparation of Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4217 Transport Assay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4218 Calculations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4219 Determination of Energy Requirement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4220 Specialized Materials Required . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4221
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Research Outlook . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4221
Mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture K. N. Timmis (ed.), Handbook of Hydrocarbon and Lipid Microbiology, DOI 10.1007/978-3-540-77587-4_329, # Springer-Verlag Berlin Heidelberg, 2010
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Measurement of Hydrocarbon Transport in Bacteria
Abstract: Hydrocarbon uptake by bacteria has not been extensively studied, and strong evidence for active transport of hydrocarbons is lacking. The volatile nature of hydrocarbons, their hydrophobicity, and their relatively low aqueous solubilities can complicate the execution of transport assays and the interpretation of the resulting data. Here we present a detailed protocol for measuring hydrocarbon uptake in bacteria, and suggest key control experiments necessary to obtain conclusive results.
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Introduction
The mechanism for intracellular accumulation of hydrocarbons by bacteria that utilize these potentially toxic compounds as carbon and energy sources is not well-established. Because hydrocarbons are naturally hydrophobic, they can freely diffuse through the cytoplasmic membrane of cells, and if unregulated can cause intracellular toxicity (reviewed in Sikkema et al., 1995) (> Chapter 52, Vol. 2, Part 9). Therefore, bacteria that use toxic molecules as sources of carbon and energy are presented with an interesting problem; cells must acquire sufficient amounts of hydrocarbons to allow growth but also manage t
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