title: Comprehensive analysis of mycolic acid subclass and molecular species composition of Mycobacterium bovis BCG Tokyo 172 cell wall skeleton (SMP-105).
abstract: The mycobacterial cell envelope consists of a characteristic cell wall skeleton (CWS), a mycoloyl arabinogalactan peptidoglycan complex, and related hydrophobic components that contribute to the cell surface properties. Since mycolic acids have recently been reported to play crucial roles in host immune response, detailed molecular characterization of mycolic acid subclasses and sub-subclasses of CWS from Mycobacterium bovis BCG Tokyo 172 (SMP-105) was performed. Mycolic acids were liberated by alkali hydrolysis from SMP-105, and their methyl esters were separated by silica gel TLC into three subclasses: alpha-, methoxy-, and keto-mycolates. Each mycolate subclass was further separated by silver nitrate (AgNO(3))-coated silica gel TLC into sub-subclasses. Molecular weights of individual mycolic acid were determined by MALDI-TOF mass spectrometry. alpha-Mycolates were sub-grouped into cis, cis-dicyclopropanoic (alpha1), and cis-monocyclopropanoic-cis-monoenoic (alpha2) series; methoxy-mycolates were sub-grouped into cis-monocyclopropanoic (m1), trans-monocyclopropanoic (m2), trans-monoenoic (m3), cis-monocyclopropanoic-trans-monoenoic (m4), cis-monoenoic (m5), and cis-monocyclopropanoic-cis-monoenoic (m6) series; and keto-mycolates were sub-grouped into cis-monocyclopropanoic (k1), trans-monocyclopropanoic (k2), trans-monoenoic (k3), cis-monoenoic (k4), and cis-monocyclopropanoic-cis-monoenoic (k5) series. The position of each functional group, including cyclopropane rings and methoxy and keto groups, was determined by analysis of the meromycolates with fast atom bombardment (FAB) mass spectrometry and FAB mass-mass spectrometry, and the cis/trans ratio of cyclopropane rings and double bonds were determined by NMR analysis of methyl mycolates. Mycolic acid subclass and molecular species composition of SMP-105 showed characteristic features including newly-identified cis-monocyclopropanoic-trans-monoenoic mycolic acid (m4).
authors.affiliation: Technology Research & Development Center, Dainippon Sumitomo Pharma Co. Ltd., 1-3-45, Kurakakiuchi, Ibaraki-shi, Osaka, Japan. yuko-uenishi@ds-pharma.co.jp
authors.forename: Yuko
authors.initials: Y
authors.lastname: Uenishi
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Fujita
authors.forename: Naoto
authors.initials: N
authors.lastname: Kusunose
authors.forename: Ikuya
authors.initials: I
authors.lastname: Yano
authors.forename: Makoto
authors.initials: M
authors.lastname: Sunagawa
mesh_terms: D017358:Cell Wall Skeleton / Q000737:chemistry* / Q000302:isolation & purification; D002855:Chromatography, Thin Layer; D006859:Hydrogen / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D015394:Molecular Structure; D009163:Mycobacterium bovis / Q000737:chemistry*; D009171:Mycolic Acids / Q000737:chemistry* / Q000302:isolation & purification; D058428:Silica Gel; D012822:Silicon Dioxide / Q000032:analysis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D017358:Cell Wall Skeleton; D009171:Mycolic Acids; D058428:Silica Gel; D012822:Silicon Dioxide; D006859:Hydrogen