Enrichment and Isolation of Hydrocarbon Degraders
Hydrocarbons are organic compounds that contain only carbon and hydrogen. They constitute the main component of crude oil, and can be divided into two classes, the saturates and the aromatics. Saturated hydrocarbons have two forms: the cyclic alkanes and
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Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3778
2 2.1 2.1.1 2.1.2 2.1.3 2.2 2.2.1 2.2.2 2.3 2.4
Experimental Approach . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3779 Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3779 Enrichment of Marine HDB . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3779 Isolation of HDB by Plate Purification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3781 Isolate Characterization and Preservation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3782 Solutions and Materials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3782 Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3782 Stock Solutions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3783 Time Considerations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3783 Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3784
K. N. Timmis (ed.), Handbook of Hydrocarbon and Lipid Microbiology, DOI 10.1007/978-3-540-77587-4_297, # Springer-Verlag Berlin Heidelberg, 2010
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Enrichment and Isolation of Hydrocarbon Degraders
Introduction
Hydrocarbons are organic compounds that contain only carbon and hydrogen. They constitute the main component of crude oil, and can be divided into two classes, the saturates and the aromatics. Saturated hydrocarbons have two forms: the cyclic alkanes and straight chain n-alkanes. The aromatic hydrocarbons have two groups varying in benzene ring number: the single-ring and more than one ring. The first group is represented by benzene, toluene, ethylbenzene and xylene (BTEX); the latter is designated as polycyclic aromatic hydrocarbon (PAH), and is represented by, e.g., naphthalene, phenanthrene, pyrene and benzo[a]pyrene, with ring numbers increasing from 2 to 5. Hydrocarbons can serve as carbon and energy sources for bacterial growth (Head et al., 2006). Bacterial degradation of various hydrocarbons has been intensively studied, and previous reviews describe this in great detail (Atlas, 1981; Leahy and Colwell, 1990; Van Hamme et al., 2003). As hydrocarbons in the environment have a wide range of distributions both spatially and temporally, it is not surprising to find hydrocarbon-degrading bacteria (HDB) to be ubiquitous over the g
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