Extracellular and surface-exposed polysaccharides of non-tuberculous mycobacteria

We studied the outermost constituents of the cell envelopes, which are involved in the interaction between the bacilli and the host cells, of five pathogenic and non-pathogenic mycobacterial species for comparison with those we have previously characterized from M. tuberculosis. The extracellular materials (ECMs) were isolated by ethanol precipitation and compared to the surface-exposed materials (SXMs) extracted by mechanical means. The materials from both sources were composed almost exclusively of polysaccharides and proteins. Two groups of mycobacteria were clearly distinguishable. The first group comprised the pathogenic species M. kansasii which produced large amounts of ECM, the glycosyl composition of which was similar to that of the SXM. The second group comprised M. avium and the nonpathogenic strains of M. gastri, M. phlei and M. smegmatis which produced small amounts of ECM. This latter group could be subdivided into those which produced carbohydrate-rich ECM (M. avium and M. gastri) and those forming protein-rich ECM (M. phlei and M. smegmatis), a classification that correlated with the difference in the growth rate of the two subgroups. The glycosyl composition of the ECM of a given species was qualitatively similar to that of the SXM, except for M. avium and M. phlei whose SXM were devoid of arabinose. In addition to glucose, mannose and arabinose, xylose was detected in the hydrolysis products of the ECM and SXM of M. smegmatis, the SXM of M. phlei and the ECM of some batches of M. avium. The polysaccharide constituents of the ECM and SXM of the different mycobacteria were purified by anion-exchange and gel-filtration chromatography; all were found to be neutral compounds devoid of acyl substituents. The extracellular polysaccharides consisted of high-molecular-mass glycogen-like glucans, arabinomannans and mannans, structurally similar to the corresponding substances previously characterized from the capsule of M. tuberculosis. The same types of polysaccharides were characterized from the SXM of all the strains, except M. avium and M. phlei which were devoid of arabinomannans. This study questions the unique and universal representation of the mycobacterial cell envelope and the existence of the so-called acidic polysaccharide-rich outer layer.