Search

The methyl esters of free mycolic acids from representative strains of Nocardia asteroides, N. brasiliensis, N. caviae and the 'rhodochrous' complex were subjected to detailed mass spectral analysis. The anhydromycolic esters of the Nocardia strains consisted of homologous series containing from zero to three double bonds, with the main components of the parent mycolic acids centred on C52 to C54 (range C46 to C58). The anhydromycolates from one rhodochrous strain, Nocardia opaca, had a molecular weight range similar to the nocardiae (C46 to C57) but the remaining rhodochrous strains gave an homologous series of anhydromycolates containing from zero to two double bonds, with the main components of the parent mycolic acids centred on C38, C42, C44 or C46 (total range from C34 to C50). The mycolic acids from the rhodochrous strains with chain lengths centred around C40 form a group intermediate in size between corynomycolic acids (centred around C32) and nocardomycolic acids (centred around C50). These data weaken the case for retaining the 'rhodochrous' complex in the genus Mycobacterium, and also show that many rhodochrous strains can be distinguished from true nocardiae and corynebacteria. These results confirm the value of lipid characters in the classification of these organisms.

Nocobactin NA, a lipid-soluble iron-chelating product with an unusual and characteristic u.v.-absorption spectrum, was isolated from Nocardia asteroides grown under conditions of iron deficiency. Its structure was determined by physical methods and by synthesis of one of its degradation products. Nocobactin NA was obtained as a homologous mixture of compounds with side chains of differing length, and resembles mycobactin M in structure except that it has an oxazole ring in place of an oxazoline ring, and the side chains in the cobactin fragment are considerably shorter.

The nocardomycolic acid compositions of extractable and the cell wall-bound lipids from five strains of Nocardia asteroides (A-23007, A-23094, B-23006, B-23095, and IFO 3384) were compared by using gas chromatography-mass spectrometry. The molecular species composition of mycolic acid differed significantly among the strains of N. asteroides. The A-23007 strain possessed the shortest species, centering at C(44(46)), and the A-23094 and IFO-3384 strains followed, each centering at C(52). The B-23006 and B-23095 strains possessed the longest species, centering at C(56) or C(54), thus indicating that N. asteroides strains accommodate a heterogeneous group in respect to carbon numbers of mycolic acids. The doublebond isomers of mycolic acids from the representative strain IFO 3384 were fully separated and analyzed by argentation thin-layer chromatography, followed by gas chromatography-mass spectrometry. The reference strain (IFO 3384) possessed up to four double bonds on the straight chain of mycolic acids ranging from C(46) to C(60). All of the species possessed a C(14) alkyl branch at C-2. The more highly unsaturated subclasses consisted of the longer-chain mycolic acids. Marked changes in mycolic acid composition were induced by altering the growth temperature of strain IFO 3384. The cells grown at the higher temperature (50 degrees C) contained more saturated mycolic acids, whereas those grown at the lower temperature (17 degrees C) had more polyunsaturated (up to tetraenoic) mycolic acids, although a significant difference in carbon chain length was not detected. These changes in the degree of unsaturation of mycolic acids occurred shortly after shifting the growth temperature from 17 to 50 degrees C at logarithmic stages of the bacterial growth, thus indicating that N. asteroides can adapt to changes in the environmental temperature by altering the structure of mycolic acids of the cell walls.

The chemical composition of the cell walls of several L-form revertants derived from Nocardia asteroides 10905 was determined at different stages of growth. It was observed that each L-form revertant had a cell well that differed from that of the parental strain when grown under identical conditions. In some strains the peptidolipid and mycolic acid components were affected the most, whereas in other strains the fatty acid, sugar, and mycolic acid moieties were altered. Shifts in mycolic acid size were prominent, whereas the basic peptidoglycan structure appeared to be affected the least. Both the method used to induce the L-form of N. asteroides 10905 and the length of time these organisms were maintained in the wall-less state affected the degree of cell wall modification during the reversion process. Thus, removal of the cell wall appeared to potentiate and select for mutational alterations within the cell envelope of N. asteroides, and these changes resulted in altered cellular and colonial morphology.

Studies were conducted on the oxidation and assimilation of n-alkyl-substituted cycloalkane substrates by several hydrocarbon-utilizing microorganisms. These microorganisms utilized heptadecylcyclohexane and dodecylcyclohexane as the sole source of carbon and energy. Neither methylcyclohexane nor ethylcyclohexane was utilized as a growth substrate by any organisms tested. Gas-liquid chromatographic analyses of fatty acids present in cells after growth on dodecylcyclohexane confirm direct incorporation of both alpha- and beta-oxidation products. Growth patterns of these organisms on n-alkyl-substituted cyclohexane fatty acids of varying chain lengths suggest a greater probability of ring cleavage when the side chain contains an odd number of carbons.