Role of lipid components in formation and reactivation of Mycobacterium smegmatis “nonculturable” cells

We have found that transition of actively dividing Mycobacterium smegmatis cells into the dormant “nonculturable” state is accompanied by increase in the protein/lipid ratio and disappearance of one of the main lipid components of the mycobacterial cells, trehalose monomycolate. In this case, oleic acid is accumulated in the culture medium due to its secretion by the mycobacterial cells. Addition of lipids of different classes to “nonculturable” M. smegmatis cells induces their resuscitation. The lipid reactivating effect is evidently caused by the presence of fatty acids in their composition, because free fatty acids also exhibited reactivation effect. Oleic acid in concentration of 0.05–3 μg/ml exhibited maximal effect, and that allows us to draw a conclusion concerning its signal role in the transition of dormant cells into active state.

[1]  A. Kaprelyants,et al.  Biochemical and morphological changes in dormant (“Nonculturable”) Mycobacterium smegmatis cells , 2010, Biochemistry (Moscow).

[2]  R. Cotter,et al.  Resuscitation of dormant Mycobacterium tuberculosis by phospholipids or specific peptides. , 2001, Biochemical and biophysical research communications.

[3]  J. Man The probability of most probable numbers , 1975, European journal of applied microbiology and biotechnology.

[4]  S. Bhaduri,et al.  Simple and rapid method for disruption of bacteria for protein studies , 1983, Applied and environmental microbiology.

[5]  H. Nikaido,et al.  The envelope of mycobacteria. , 1995, Annual review of biochemistry.

[6]  Christopher Dye,et al.  Global Burden of Tuberculosis: Estimated Incidence, Prevalence, and Mortality by Country , 1999 .

[7]  D. Kell,et al.  Adoption of the transiently non-culturable state--a bacterial survival strategy? , 2003, Advances in microbial physiology.

[8]  A. Kaprelyants,et al.  The effect of liposomes on the growth and sensitivity of Mycobacterium smegmatis to isoniazide , 2007, Applied Biochemistry and Microbiology.

[9]  J. M. Dow,et al.  A novel regulatory system required for pathogenicity of Xanthomonas campestris is mediated by a small diffusible signal molecule , 1997, Molecular microbiology.

[10]  C. Dye,et al.  Consensus statement. Global burden of tuberculosis: estimated incidence, prevalence, and mortality by country. WHO Global Surveillance and Monitoring Project. , 1999, JAMA.

[11]  Markus R. Wenk,et al.  Triacylglycerol Utilization Is Required for Regrowth of In Vitro Hypoxic Nonreplicating Mycobacterium bovis Bacillus Calmette-Guerin , 2009, Journal of bacteriology.

[12]  C. Senner,et al.  Cytological and Transcript Analyses Reveal Fat and Lazy Persister-Like Bacilli in Tuberculous Sputum , 2008, PLoS medicine.

[13]  F. R. Cox,et al.  Rapid Tween 80 hydrolysis test for mycobacteria , 1978, Journal of clinical microbiology.

[14]  B. Abomoelak,et al.  A Novel In Vitro Multiple-Stress Dormancy Model for Mycobacterium tuberculosis Generates a Lipid-Loaded, Drug-Tolerant, Dormant Pathogen , 2009, PloS one.

[15]  M. Young,et al.  Formation of 'non-culturable' cells of Mycobacterium smegmatis in stationary phase in response to growth under suboptimal conditions and their Rpf-mediated resuscitation. , 2004, Microbiology.

[16]  Pontus Larsson,et al.  Sporulation in mycobacteria , 2009, Proceedings of the National Academy of Sciences.

[17]  J. M. Dow,et al.  Diffusible signals and interspecies communication in bacteria. , 2008, Microbiology.

[18]  N. Connell Mycobacterium: isolation, maintenance, transformation, and mutant selection. , 1994 .

[19]  Daniel F Lusche,et al.  Arachidonic acid is a chemoattractant for Dictyostelium discoideum cells , 2007, Journal of Biosciences.

[20]  J. Maeda,et al.  Isolation and Biochemical Activities of Trehalose-6-Monomycolate of Mycobacterium tuberculosis , 1974, Infection and immunity.

[21]  E. L. Armstrong,et al.  Synthesis of trehalose dimycolate (cord factor) by a cell-free system of Mycobacterium smegmatis. , 1982, Biochemical and biophysical research communications.

[22]  T. Parish Mycobacterium molecular microbiology , 2005 .

[23]  L. G. Wayne Dormancy ofMycobacterium tuberculosis and latency of disease , 1994, European Journal of Clinical Microbiology and Infectious Diseases.