Analysis of respiratory quinones
For this method 200 mg freeze dried cells are needed. If the analysis of polar lipids is ordered in parallel we need 200 mg freeze dried cells in total for both methods.
Extraction of respiratory lipoquinones and polar lipids
Respiratory lipoquinones are extracted from 100 mg of freeze dried cell material using the two stage method described by Tindall (1990a; 1990b). Respiratory quinones are extracted using methanol:hexane (Tindall, 1990a, 1990b), followed by phase separation into hexane.
Analysis of respiratory lipoquinones
Respiratory lipoquinones are separated into their different classes (menaquinones, ubiquinones, etc.) by thin layer chromatography on silica gel (Macherey-Nagel Art. No. 805 023), using hexane:tert¬butylmethylether (9:1 v/v) as solvent. UV absorbing bands corresponding to the different quinone clases (e.g. menaquinones or ubiquinones) were removed from the plate and further analysed by HPLC. This step is carried out on a LDC Analytical (Thermo Separation Products) HPLC fitted with a reverse phase column (Macherey-Nagel, 2 mm x 125 mm, 3 µm, RP18) using methanol:heptane 9:1 (v/v) as the eluant. Respiratory lipoquinones are detected at 269 nm.
- Tindall, B.J. (1990a). A comparative study of the lipid composition of Halobacterium saccharovorum from various sources. Syst. Appl. Microbiol. 13, 128-130
- Tindall, B.J. (1990b). Lipid composition of Halobacterium lacusprofundi. FEMS Microbiol. Letts. 66, 199-202
Coordination of the identification services. Identification of aerobic and facultative anaerobic Gram positive and Gram negative bacteria, physiological, biochemical and morphological characterization. Curator for filamentous bacteria belonging to different lines of descent (e.g. Beggiatoa, Thiothrix, Flexibacter, Sphaerotilus)