Gas chromatography of mycobacterial fatty acids and alcohols: Diagnostic applications

Abstract

Jantzen, E., Tangen, T. & Eng. J. Gas chromatography of mycobacterial fatty acids and alcohols: Diagnostic applications. APMIS 5'7: [1037][1038][1039][1040][1041][1042][1043][1044][1045] 1989.Capillary gas chromatography of cellular fatty acids and alcohols has been used as a routine method for a period of two years in the mycobacterial diagnostic laboratory of Statens institutt for folkehelse, Oslo, Norway. All mycobacteria (165 isolates) other than Mycobacterium tuberculosis (MOTT) and 24 randomly selected M . tzib~~rczilosis isolates were studied. Twelve characteristic lipid constituents allowed the construction of a diagnostic scheme. Without exceptions. all 36 examined isolates belonging to the M. ttthc,r~.zf,uk)sis-complex were characterized by a relatively high concentration level of hexacosanoic acid (mean: 4%. range: I-l3%), low level of tetracosanoic acid (mean: 1%. range: 0.1-3%), lack of methylbranched acids other than tuberculostearic acid, and lack of fatty alcohols. Members of the MAIS-complex (73 isolates) were all characterized by the general presence of the fatty alcohols 2-octadecanol (mean: 2%, range: 0.1-5%) and 2-eicosanol (mean: 7%, range: 2-2 1 %), relatively high levels of tetracosanoic acid (mean: 5%, range: 1-15%) and lack (or trace) of hexacosanoic acid and methylbranched acids other than tuberculostearic acid. All 16 isolates of M . gordonae were easily recognized by their unique lack of tuberculostearic acid and their content of 2-methyl-tetradecanoic acid (mean: 5%, range: 2-1 2%), and the M . xenopi isolates were the only examined strains containing the fatty alcohol 2-docosanol (mean: 9%, range: 2-13%). The six M . mulmoense strains contained the two unique constituents 2-methyl eicosanoic acid (mean: 39/0, range: 1-4%) and 2,4,6-trimethyl tetracosanoic acid (mean: 3%, range: 2-4%). The ten strains of M . kunsusii were characterized by 2.4-dimethyl tetradecanoic acid (mean: 5%, range: 1-1 I%), whereas the seven strains of M . rriarinum shared 2,4-dimethyl hexadecanoic acid (mean: 4%, range 0.2-12%) as a specific marker.

LosA, a Key Glycosyltransferase Involved in the Biosynthesis of a Novel Family of Glycosylated Acyltrehalose Lipooligosaccharides from Mycobacterium marinum

“…This is in line with the recent demonstration that strains of M. tuberculosis deficient in the production of phenolphthiocerol derivatives (which are precursors of PGLs) are natural mutants with a frameshift mutation in pks15/1 (39). A single open reading frame for pks15/1 is found in Mycobacterium bovis BCG, M. leprae, and the Canetti strains of M. tuberculosis, which produce PGLs (36). The presence of LOS in the Canetti strain could be explained by the assumption that M. tuberculosis Canetti is either the common ancestor for the M. tuberculosis strains or it diverged before the appearance of other members of the M. tuberculosis complex (40).…”

“…The absolute stereochemistry of these branched chain acids also remains to be determined; the availability of synthetic routes to such 2,4-dimethyl branched acids (34) will facilitate this. The 2,4-dimethylhexadecanoate was found previously (35,36) to be a characteristic biomarker fatty acid for M. marinum and this study shows the natural location of this acid, for the first time.…”

Applications of cellular fatty acid analysis

“…Because of the abundance of mycolic acids in mycobacterial species, several studies have addressed the separation of compounds of both types, i.e., those with chains of >20 carbons (mycolic acid cleavage products) and those with chains of <20 carbons (constitutive fatty acids) (58,87,150). Quantitative analysis of fatty acids belonging to these two groups results in accurate identification to the species level (58,69,82,150,151 Vibrionaceae. The Vibrionaceae may be distinguished from the Enterobacteriaceae by the presence of significant amounts of branched-chain CFAs, especially iso-C16.0 and iso-C18:0, in addition to greater amounts of C16:1 (9).…”

Characterization of Mycobacterium bohemicum Isolated from Human, Veterinary, and Environmental Sources

“…The isolates were cultured on mycobacteria 7H11 agar supplemented with Middlebrook OADC enrichment (Difco, Detroit, Mich.) at 30°C (environmental isolates) or 36°C (clinical isolates). Fatty acid methyl esters were produced by acid methanolysis (5). Analyses of lipid composition, including fatty acids, fatty alcohols, and mycolic acid cleavage products, were performed using a Perkin-Elmer (Norwalk, Conn.) AutoSystem gas chromatograph.…”