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

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 propos...

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Autores principales: Singh, Albel, Varela, Cristian, Bhatt, Kiranmai, Veerapen, Natacha, Lee, Oona Y. C., Wu, Houdini H. T., Besra, Gurdyal S., Minnikin, David E., Fujiwara, Nagatoshi, Teramoto, Kanae, Bhatt, Apoorva
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5065219/
https://www.ncbi.nlm.nih.gov/pubmed/27741286
http://dx.doi.org/10.1371/journal.pone.0164253
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author Singh, Albel
Varela, Cristian
Bhatt, Kiranmai
Veerapen, Natacha
Lee, Oona Y. C.
Wu, Houdini H. T.
Besra, Gurdyal S.
Minnikin, David E.
Fujiwara, Nagatoshi
Teramoto, Kanae
Bhatt, Apoorva
author_facet Singh, Albel
Varela, Cristian
Bhatt, Kiranmai
Veerapen, Natacha
Lee, Oona Y. C.
Wu, Houdini H. T.
Besra, Gurdyal S.
Minnikin, David E.
Fujiwara, Nagatoshi
Teramoto, Kanae
Bhatt, Apoorva
author_sort Singh, Albel
collection PubMed
description 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 ~C(26)-C(48) 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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spelling pubmed-50652192016-10-27 Identification of a Desaturase Involved in Mycolic Acid Biosynthesis in Mycobacterium smegmatis Singh, Albel Varela, Cristian Bhatt, Kiranmai Veerapen, Natacha Lee, Oona Y. C. Wu, Houdini H. T. Besra, Gurdyal S. Minnikin, David E. Fujiwara, Nagatoshi Teramoto, Kanae Bhatt, Apoorva PLoS One Research Article 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 ~C(26)-C(48) 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. Public Library of Science 2016-10-14 /pmc/articles/PMC5065219/ /pubmed/27741286 http://dx.doi.org/10.1371/journal.pone.0164253 Text en © 2016 Singh et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Singh, Albel
Varela, Cristian
Bhatt, Kiranmai
Veerapen, Natacha
Lee, Oona Y. C.
Wu, Houdini H. T.
Besra, Gurdyal S.
Minnikin, David E.
Fujiwara, Nagatoshi
Teramoto, Kanae
Bhatt, Apoorva
Identification of a Desaturase Involved in Mycolic Acid Biosynthesis in Mycobacterium smegmatis
title Identification of a Desaturase Involved in Mycolic Acid Biosynthesis in Mycobacterium smegmatis
title_full Identification of a Desaturase Involved in Mycolic Acid Biosynthesis in Mycobacterium smegmatis
title_fullStr Identification of a Desaturase Involved in Mycolic Acid Biosynthesis in Mycobacterium smegmatis
title_full_unstemmed Identification of a Desaturase Involved in Mycolic Acid Biosynthesis in Mycobacterium smegmatis
title_short Identification of a Desaturase Involved in Mycolic Acid Biosynthesis in Mycobacterium smegmatis
title_sort identification of a desaturase involved in mycolic acid biosynthesis in mycobacterium smegmatis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5065219/
https://www.ncbi.nlm.nih.gov/pubmed/27741286
http://dx.doi.org/10.1371/journal.pone.0164253
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