Novel O-Methylated Terminal Glucuronic Acid Characterizes the Polar Glycopeptidolipids of Mycobacterium habana Strain TMC 5135

Abstract

Mycobacterium "habana" strain TMC 5135, which has been proposed as a vaccine against both leprosy and tuberculosis, is considered to be a strain of serotype I of the recognized species Mycobacterium simiae. We have now shown that each of these strains possesses characteristic polar glycopeptidolipids (GPL) which are sufficiently different to allow unequivocal strain identification. Thin layer chromatographic analysis demonstrated that M. habana synthesizes a family of apolar GPLs and three distinct polar GPLs (pGPL-I to -III) which exhibited migration patterns different from those of M. simiae serotype I (pGPL-Sim). Using a combination of chemical, mass spectrometric, and proton-NMR analyses, the GPLs from M. habana were determined to be based on the same generic structure as those from the M.

Analysis of the structure of mycolic acids of Mycobacterium simiae reveals a particular composition of α-mycolates in strain ‘habana’ TMC 5135, considered as immunogenic in tuberculosis and leprosy

“…Dried cells (0.5-1.0 g) were extracted overnight at room temperature with ethanol/ diethyl ether (1 : 1, v/v), filtered, and re-extracted with chloroform/ methanol (1 : 1, v/v) under the same conditions (Khoo et al, 1996). Extractable lipids were employed for polar glycopeptidolipid analysis (data not shown).…”

“…habana' TMC 5135 has been reported (Khoo et al, 1996), as well as the general composition of lipids of several strains within the 'habana' group (Mederos et al, 1998), and this has taxonomic potential. However, the analysis of the fine structure of the mycolic acids of M. simiae is still pending.…”

Synthesis of an unusual polar glycopeptidolipid in glucose-limited culture of Mycobacterium smegmatis

“…MALDI-TOF (matrix assisted laser desorption ionization-time of flight ; Kratos) was used to obtain the mass of each GPL species of M. smegmatis in positive ion mode using gentisic acid as the matrix. For further analysis of structure, GPLs were permethylated as described earlier (Khoo et al 1996). Briefly, 0n5 ml DMSO was mixed with one pellet NaOH and the slurry was added to a vial containing 500 µg GPL.…”

Identification and Characterization of Two Novel Methyltransferase Genes That Determine the Serotype 12-Specific Structure of Glycopeptidolipids of Mycobacterium intracellulare

“…GPL 7 and GPL 12 were purified from M. intracellulare NF 027 and NF 103, respectively. The preparation of GPLs was performed as described previously (10,15,17). Briefly, each strain was grown in Middlebrook 7H9 broth (Difco Laboratories, Detroit, MI) with 0.5% glycerol and 10% Middlebrook oleic acid-albumin-dextrose-catalase enrichment (Difco Laboratories) at 37°C for 2 to 3 weeks.…”