Characterization of Mycolic Acids in Total Fatty Acid Methyl Ester Fractions from &lt;i&gt;Mycobacterium&lt;/i&gt; Species by High Resolution MALDI-TOFMS

Teramoto, Kanae; Suga, Mitsuo; Sato, Takafumi; Wada, Takayuki; Yamamoto, A.; Fujiwara, Nagatoshi

doi:10.5702/massspectrometry.a0035

Cited by 14 publications

(11 citation statements)

References 32 publications

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“…S1C), so the mycolic acid representation of the MWE is consistent with the overall mycolic acid composition of the bacterium. The mycolic acid composition of our biofilm-grown bacteria is consistent with what others have reported recently for H37Rv cultured on Ogawa agar, although we acknowledge that different growth conditions can change the mycolic acid profiles of mycobacteria (29).…”

Section: Resultssupporting

confidence: 78%

The Mycobacterium tuberculosis MmpL11 Cell Wall Lipid Transporter Is Important for Biofilm Formation, Intracellular Growth, and Nonreplicating Persistence

et al. 2017

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The mycobacterial cell wall is crucial to the host-pathogen interface, because it provides a barrier against antibiotics and the host immune response. In addition, cell wall lipids are mycobacterial virulence factors. The mycobacterial membrane protein large (MmpL) proteins are cell wall lipid transporters that are important for basic mycobacterial physiology and Mycobacterium tuberculosis pathogenesis. MmpL3 and MmpL11 are conserved across pathogenic and nonpathogenic mycobacteria, a feature consistent with an important role in the basic physiology of the bacterium. MmpL3 is essential and transports trehalose monomycolate to the mycobacterial surface. In this report, we characterize the role of MmpL11 in M. tuberculosis. M. tuberculosis mmpL11 mutants have altered biofilms associated with lower levels of mycolic acid wax ester and long-chain triacylglycerols than those for wild-type bacteria. While the growth rate of the mmpL11 mutant is similar to that of wild-type M. tuberculosis in macrophages, the mutant exhibits impaired survival in an in vitro granuloma model. Finally, we show that the survival or recovery of the mmpL11 mutant is impaired when it is incubated under conditions of nutrient and oxygen starvation. Our results suggest that MmpL11 and its cell wall lipid substrates are important for survival in the context of adaptive immune pressure and for nonreplicating persistence, both of which are critically important aspects of M. tuberculosis pathogenicity.

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Section: Resultssupporting

confidence: 78%

The Mycobacterium tuberculosis MmpL11 Cell Wall Lipid Transporter Is Important for Biofilm Formation, Intracellular Growth, and Nonreplicating Persistence

et al. 2017

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“…We further analysed total mycolates extracted from cultures of Δ MsdesA1 grown in the presence or absence of acetamide, using MALDI-TOF MS with a spiral trajectory [ 23 , 24 ]. We chose this method over conventional MALDI-TOS MS as it gives high mass resolution and accuracy, and enables the separation of adjacent peaks with a 0.036 Da mass difference.…”

Section: Resultsmentioning

confidence: 99%

Identification of a Desaturase Involved in Mycolic Acid Biosynthesis in Mycobacterium smegmatis

et al. 2016

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Mycolic acids are unique long chain fatty acids found in the cell walls of mycobacteria including the tubercle bacillus, Mycobacterium tuberculosis. The introduction of double bonds in mycolic acids remains poorly understood, however, genes encoding two potential aerobic desaturases have been proposed to be involved in this process. Here we show that one of these genes, desA1, is essential for growth of the saprophytic Mycobacterium smegmatis. Depletion of desA1 in a M. smegmatis conditional mutant led to reduction of mycolic acid biosynthesis and loss of viability. The DesA1-depleted cells exhibited two other phenotypes: using 14[C]-labelling, we detected the accumulation of minor mycolic acid-related species that migrated faster in a silver TLC plate. Spiral Time of Flight Mass Spectroscopic analysis suggested the presence of species with sizes corresponding to what were likely monoenoic derivatives of α-mycolic acids. Additionally, conditional depletion led to the presence of free fatty acyl species of lengths ~C26-C48 in the lysing cells. Cell viability could be rescued in the conditional mutant by Mycobacterium tuberculosis desA1, highlighting the potential of desA1 as a new drug target in pathogenic mycobacteria.

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“…Visualized spots of total lipids, including mycolic acids, showed almost identical patterns in TLC analysis, indicating the conservation of the lipid layer in the cell membrane. The extracted fatty acid methylesters were further analyzed by MALDI TOF mass spectrometry with a spiral ion trajectory (MALDI spiral TOF-MS) [ 45 , 46 ]. Mycolic acid fractions showed identical patterns between formaldehyde-fixed and non-fixed cells ( Fig 1D ).…”

Section: Resultsmentioning

confidence: 99%

Killing of Mycolic Acid-Containing Bacteria Aborted Induction of Antibiotic Production by Streptomyces in Combined-Culture

et al. 2015

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Co-culture of Streptomyces with mycolic acid-containing bacteria (MACB), which we termed “combined-culture,” alters the secondary metabolism pattern in Streptomyces and has been a useful method for the discovery of bioactive natural products. In the course of our investigation to identify the inducing factor(s) of MACB, we previously observed that production of pigments in Streptomyces lividans was not induced by factors such as culture extracts or mycolic acids. Although dynamic changes occurred in culture conditions because of MACB, the activation of pigment production by S. lividans was observed in a limited area where both colonies were in direct contact. This suggested that direct attachment of cells is a requirement and that components on the MACB cell membrane may play an important role in the response by S. lividans. Here we examined whether this response was influenced by dead MACB that possess intact mycolic acids assembled on the outer cell membrane. Formaldehyde fixation and γ-irradiation were used to prepare dead cells that retain their shape and mycolic acids of three MACB species: Tsukamurella pulmonis, Rhodococcus erythropolis, and Rhodococcus opacus. Culturing tests verified that S. lividans does not respond to the intact dead cells of three MACB. Observation of combined-culture by scanning electron microscopy (SEM) indicated that adhesion of live MACB to S. lividans mycelia were a significant interaction that resulted in formation of co-aggregation. In contrast, in the SEM analysis, dead cells were not observed to adhere. Therefore, direct attachment by live MACB cells is proposed as one of the possible factors that causes Streptomyces to alter its specialized metabolism in combined-culture.

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Characterization of Mycolic Acids in Total Fatty Acid Methyl Ester Fractions from <i>Mycobacterium</i> Species by High Resolution MALDI-TOFMS

Cited by 14 publications

References 32 publications

The Mycobacterium tuberculosis MmpL11 Cell Wall Lipid Transporter Is Important for Biofilm Formation, Intracellular Growth, and Nonreplicating Persistence

The Mycobacterium tuberculosis MmpL11 Cell Wall Lipid Transporter Is Important for Biofilm Formation, Intracellular Growth, and Nonreplicating Persistence

Identification of a Desaturase Involved in Mycolic Acid Biosynthesis in Mycobacterium smegmatis

Killing of Mycolic Acid-Containing Bacteria Aborted Induction of Antibiotic Production by Streptomyces in Combined-Culture

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