1887

Abstract

The lipid-rich , to which and species belong, produce both fatty acids and mycolic acids. Compared with most other bacteria, possesses two fatty acid synthases, encoded by (8907 kb; FAS-IA) and (8988 kb; FAS-IB). Here, it was shown by mutational analyses that is essential but is not. However, in a background, the mutation results in a slightly reduced growth yield, -glutamate production is increased, and comparative lipid analysis suggests that FAS-IB is active primarily to supply palmitate. Transcript quantifications revealed that the transcript contributes 32 % to both transcripts during growth on glucose, affirmative for expression, and that is subordinate to . The transcript is downregulated by 8·3-fold during growth on acetate as compared with glucose. The lipid analyses also demonstrate that cells grown on propionate produce a number of uneven fatty acids (e.g. 15 : 0, 17 : 0, 17 : 1), which are not present in cells grown on glucose or acetate, suggesting that fatty acid synthase may also use propionyl-CoA as the priming unit in fatty acid synthesis. The fatty acid auxotrophic double mutant was used to determine the suggested incorporation of fatty acids into mycolic acids. Supplementation of this mutant with uniformly labelled [C]oleate and analysis of isolated mycolic acids confirmed that mature mycolic acids in the mutant consist exclusively of two fused [C]oleate molecules. In addition to an altered phospholipid profile, the mutant also exhibits differences in its mycolic acid profile. Taken together, the results show that although FAS-IA is the most relevant fatty acid synthase of and FAS-IB is supplementary, both synthases are necessary to produce the characteristic lipid environment of this organism.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.28012-0
2005-07-01
2024-05-04
Loading full text...

Full text loading...

/deliver/fulltext/micro/151/7/mic1512421.html?itemId=/content/journal/micro/10.1099/mic.0.28012-0&mimeType=html&fmt=ahah

References

  1. Claes W. A., Pühler A., Kalinowski J. 2002; Identification of two prpDBC gene clusters in Corynebacterium glutamicum and their involvement in propionate degradation via the 2-methylcitrate cycle. J Bacteriol 184:2728–2739 [CrossRef]
    [Google Scholar]
  2. Cole S. T., Brosch R., Parkhill J. 39 other authors 1998; Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence. Nature 393:537–544 [CrossRef]
    [Google Scholar]
  3. Collins M. D., Goodfellow M., Minnikin D. E. 1982; Fatty acid composition of some mycolic acid-containing coryneform bacteria. J Gen Microbiol 128:2503–2509
    [Google Scholar]
  4. Cronan J. E. Jr, Rock C. O. 1996; Biosynthesis of membrane lipids. In Escherichia coli and Salmonella: Cellular and Molecular Biology pp 612–636 Edited by Neidhardt F. C. others Washington, DC: American Society for Microbiology;
    [Google Scholar]
  5. Eggeling L., Bott M. 2005 Handbook of Corynebacterium glutamicum Boca Raton, FL: CRC Press;
    [Google Scholar]
  6. Eggeling L., Sahm H. 1999; l-glutamate and l-lysine: traditional products with impetuous developments. Appl Microbiol Biotechnol 52:146–153 [CrossRef]
    [Google Scholar]
  7. Eggeling L., Sahm H. 2001; The cell wall barrier of Corynebacterium glutamicum and amino acid efflux. J Biosci Bioeng 92:201–213 [CrossRef]
    [Google Scholar]
  8. Gande R., Gibson K. J. C., Brown A. K. 7 other authors 2004; Acyl-CoA carboxylases ( accD2 and accD3 ), together with a unique polyketide synthase (Cg- pks ), are key to mycolic acid biosynthesis in Corynebacterianeae such as Corynebacterium glutamicum and Mycobacterium tuberculosis . J Biol Chem 279:44847–44857 [CrossRef]
    [Google Scholar]
  9. Hoischen C., Krämer R. 1990; Membrane alteration is necessary but not sufficient for effective glutamate secretion in Corynebacterium glutamicum . J Bacteriol 172:3409–3416
    [Google Scholar]
  10. Jarlier V., Nikaido H. 1994; Mycobacterial cell wall: structure and role in natural resistance to antibiotics. FEMS Microbiol Lett 123:11–18 [CrossRef]
    [Google Scholar]
  11. Kalinowski J., Bathe B., Bartels D. 24 other authors 2003; The complete Corynebacterium glutamicum ATCC 13032 genome sequence and its impact on the production of l-aspartate-derived amino acids and vitamins. J Biotechnol 104:5–25 [CrossRef]
    [Google Scholar]
  12. Kawaguchi A., Seyama Y., Yamakawa T., Okuda S. 1981; Fatty acid synthesis from Brevibacterium ammoniagenes . Methods Enzymol 71:120–127
    [Google Scholar]
  13. Klatte S., Kroppenstedt R. M., Rainey F. A. 1994; Rhodococcus opacus sp-nov, an unusual nutritionally versatile Rhodococcus species. Syst Appl Microbiol 17:355–360 [CrossRef]
    [Google Scholar]
  14. Liu J., Nikaido H. 1999; A mutant of Mycobacterium smegmatis defective in the biosynthesis of mycolic acids accumulates meromycolates. Proc Natl Acad Sci U S A 96:4011–4016 [CrossRef]
    [Google Scholar]
  15. Mailaender C., Reiling N., Engelhardt H., Bossmann S., Ehlers S., Niederweis M. 2004; The MspA porin promotes growth and increases antibiotic susceptibility of both Mycobacterium bovis BCG and Mycobacterium tuberculosis . Microbiology 150:853–864 [CrossRef]
    [Google Scholar]
  16. Nampoothiri K. M., Hoischen C., Bathe B., Krumbach K., Sahm H., Eggeling L, Möckel B., Pfefferle W. 2002; Expression of genes of lipid synthesis and altered lipid composition modulates l-glutamate efflux of Corynebacterium glutamicum . Appl Microbiol Biotechnol 58:89–96 [CrossRef]
    [Google Scholar]
  17. Okazaki H., Kanzaki T., Doi M., Sumino Y., Fukuda H. 1967; l-Glutamic acid fermentation. II. The production of l-glutamic acid by an oleic-acid requiring mutant. Agric Biol Chem 31:1314–1317 [CrossRef]
    [Google Scholar]
  18. Portevin D., De Sousa-D'Auria C., Houssin C., Grimaldi C., Chami M., Daffe M., Guilhot C. 2004; A polyketide synthase catalyzes the last condensation step of mycolic acid biosynthesis in mycobacteria and related organisms. Proc Natl Acad Sci U S A 101:314–319 [CrossRef]
    [Google Scholar]
  19. Schäfer A., Tauch A., Jäger W., Kalinowski J., Thierbach G., Pühler A. 1994; Small mobilizable multi-purpose cloning vectors derived from the Escherichia coli plasmids pK18 and pK19: selection of defined deletions in the chromosome of Corynebacterium glutamicum . Gene 145:69–73 [CrossRef]
    [Google Scholar]
  20. Stackebrandt E., Rainey F. A., Ward-Rainey N. L. 1997; Proposal for a new hierachic classification system, Actinobacteria classis nov. Int J Syst Bacteriol 47:479–491 [CrossRef]
    [Google Scholar]
  21. Stuible H., Wagner C., Andreou I., Huter G., Haselmann J., Schweizer E. 1996; Identification and functional differentiation of two type I fatty acid synthases in Brevibacterium ammoniagenes . J Bacteriol 178:4787–4793
    [Google Scholar]
  22. Stuible H. P., Meurer G., Schweizer E. 1997; Heterologous expression and biochemical characterization of two functionally different type I fatty acid synthases from Brevibacterium ammoniagenes . Eur J Biochem 247:268–273 [CrossRef]
    [Google Scholar]
  23. Takayama K., Wang C., Besra G. S. 2005; Pathway to synthesis and processing of mycolic acids in Mycobacterium tuberculosis . Clin Microbiol Rev 18:81–101 [CrossRef]
    [Google Scholar]
  24. Wendisch V. F., Spies M., Reinscheid D. J., Schnicke S., Sahm H., Eikmanns B. J. 1997; Regulation of acetate metabolism in Corynebacterium glutamicum : transcriptional control of the isocitrate lyase and malate synthase genes. Arch Microbiol 168:262–269 [CrossRef]
    [Google Scholar]
  25. Wendisch V. F., Zimmer D. P., Khodursky A., Peter B., Cozzarelli N., Kustu S. 2001; Isolation of Escherichia coli mRNA and comparison of expression using mRNA and total RNA on DNA microarrays. Anal Biochem 290:205–213 [CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.28012-0
Loading
/content/journal/micro/10.1099/mic.0.28012-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error