Two-dimensional thin-layer chromatography of whole-organism acid methanolysates of Mycobacterium chelonei gives a characteristic pattern of two non-polar mycolic acid methyl esters which allows the organism to be distinguished from all other mycobacteria including Mycobacterium fortuitum. The mycolic acids from Mycobacterium chelonei were composed of approximately equal amounts of a diunsaturated alpha-mycolate and a lower molecular weight alpha'-mycolate, though minor amounts of a different alpha-mycolate were also detected. These mycolic acids are the first examples of natural mixtures from mycobacteria lacking major amounts of acids having oxygen functions in addition to the 3-hydroxy acid unit.
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