Mycolic acid structure determines the fluidity of the mycobacterial cell wall

The low permeability of the mycobacterial cell wall is thought to contribute to the well known resistance of mycobacteria to antibiotics and chemotherapeutic agents. We have used differential scanning calorimetry to demonstrate that the high temperature phase transition observed in purified cell walls, usually in the 6070 °C range, suggestive of a lipid environment of extremely low fluidity, can also be observed in whole organisms and in cell walls from which much of the free lipids was removed by extraction with Triton X-114. A survey of seven mycobacterial species demonstrated that this high temperature transition was a general property of these organisms. Cell walls isolated from two Corynebacterium species, which contain much shorter corynemycolic acids, displayed a much lower temperature transition, suggesting that the transition temperature was directly correlated to the length of mycolic acid. Methyl esters of mycolic acids were found to have a phase transition temperature that was linearly related to the amount of trans-mycolate. Both Mycobacterium avium and M. smegmatis responded to increasing growth temperature by increasing the proportion of trans-mycolate and displaying a correspondingly higher melting temperature. Whole cells of M. smegmatis grown at higher temperature allowed a less rapid influx of two lipophilic agents, norfloxacin and chenodeoxycholate. These results provide strong evidence that the nature of mycolic acid plays a crucial role in determining the fluidity and permeability of mycobacterial cell wall.