Optimization of Coprogen Production in Neurospora crassa

Abstract

Coprogen production of Neurospora crassa was dependent on glucose, aspartate and iron contents as well as on initial pH of the culture media. Surplus iron and acidic pH hindered the production of coprogen as well as the transcription of the sid1 gene (NCU07117) encoding putative L-ornithine-N5-monooxygenase, the first enzyme in the coprogen biosynthetic pathway. High glucose (40 g/l) and aspartate (21 g/l) concentrations were beneficial for coprogen synthesis, but neither glucose nor aspartate affected the sid1 transcription. Moreover, efficient coprogen production was observed after glucose had been consumed, which suggested that N. crassa accumulated iron even in non-growing, carbon-starving cultures.

References

  1. 1.

    Balla, J., Vercelotti, G. M., Jeney, V., Yachie, A., Varga, Z., Eaton, J. W., Balla, G. (2005) Heme, heme oxygenase and ferritin. Mol. Nutr. Food Res. 49, 1030–1043.

    CAS  Article  Google Scholar 

  2. 2.

    Charlang, G., Ng, B., Horowitz, N. H., Horowitz, R. M. (1981) Cellular and extracellular siderophores of Aspergillus nidulans and Penicillium chrysogenum. Mol. Cell Biol. 1, 94–100.

    CAS  Article  Google Scholar 

  3. 3.

    Enyedy, É. A., Pócsi, I., Farkas, E. (2004) Complexation of desferricoprogen with trivalent Fe, A., Ga, In and divalent Fe, N., Cu, Zn metal ions: effects of the linking chain structure on the metal binding ability of hydroxamate based siderophores. J. Inorg. Biochem. 98, 1957–1966.

    CAS  Article  Google Scholar 

  4. 4.

    Haas, H. (2003) Molecular genetics of fungal siderophore biosynthesis and uptake: the role of siderophores in iron uptake and storage. Appl. Microbiol. Biotechnol. 62, 316–330.

    CAS  Article  Google Scholar 

  5. 5.

    Hof, C., Eisfeld, K., Welzel, K., Antelo, L., Foster, A. J., Anke, H. (2007) Ferricrocin synthesis in Magnaporthe grisea and its role in pathogenicity in rice. Mol. Plant. Pathol. 8, 163–172.

    CAS  Article  Google Scholar 

  6. 6.

    Hördt, W., Römheld, V., Winkelmann, G. (2000) Fusarinines and dimerum acid, mono- and dihydroxamate siderophores from Penicillium chrysogenum, improve iron utilization by strategy I and strategy II plants. BioMetals 13, 37–47.

    Article  Google Scholar 

  7. 7.

    Horowitz, N. H., Charlang, G., Horn, G., Williams, N. P. (1976) Isolation and identification of the conidial germination factor of Neurospora crassa. J. Bacteriol. 127, 135–140.

    CAS  PubMed  PubMed Central  Google Scholar 

  8. 8.

    Howard, D. H. (1999) Acquisition, transport, and storage of iron by pathogenic fungi. Clin. Microbiol. Rev. 12, 394–404.

    CAS  Article  Google Scholar 

  9. 9.

    Huschka, H., Naegeli, H. U., Leuenberger-Ryf, H., Keller-Schierlein, W., Winkelmann, G. (1985) Evidence for a common siderophore transport system but different siderophore receptors in Neurospora crassa. J. Bacteriol. 162, 715–721.

    CAS  PubMed  PubMed Central  Google Scholar 

  10. 10.

    John, S. G., Ruggiero, C. E., Hersman, L. E., Tung, C. S., Neu, M. P. (2001) Siderophore mediated plutonium accumulation by Microbacterium flavescens (JG-9). Environ. Sci. Technol. 35, 2942–2948.

    CAS  Article  Google Scholar 

  11. 11.

    Konetschny-Rapp, S., Huschka, H. G., Winkelmann, G., Jung, G. (1988) High-performance liquid chromatography of siderophores from fungi. Biol. Met. 1, 9–17.

    CAS  Article  Google Scholar 

  12. 12.

    Leary, N. O., Pembroke, A., Duggan, P. F. (1992) Improving accuracy of glucose oxidase procedure for glucose determinations on discrete analyzers. Clin. Chem. 38, 298–302.

    CAS  PubMed  Google Scholar 

  13. 13.

    Lee, S. L., Chen, W. C. (1997) Optimization of medium composition for the production of glucosyltransferase by Aspergillus niger with response surface methodology. Enzyme Microbiol. Technol. 21, 436–440.

    CAS  Article  Google Scholar 

  14. 14.

    Matzanke, B. F., Bill, E., Trautwein, A. X., Winkelmann, G. (1988) Ferricrocin functions as the main intracellular iron-storage compound in mycelia of Neurospora crassa. Biol. Metals 1, 18–25.

    CAS  Article  Google Scholar 

  15. 15.

    Pócsi, I., Jeney, V., Kertai, P., Pócsi, I., Emri, T., Gyémánt, Gy., Fésüs, L., Balla, J., Balla, Gy. (2008) Fungal siderophores function as protective agents of LDL oxidation and are promising anti-atherosclerotic metabolites in functional food. Mol. Nutr. Food Res. 52, 1434–1447.

    Article  Google Scholar 

  16. 16.

    Richardson, D. R. (2002) Iron chelators as therapeutic agents for the treatment of cancer. Crit. Rev. Oncol/Hematol. 42, 267–281.

    CAS  Article  Google Scholar 

  17. 17.

    Skromne, I., Sanchez, O., Aguierre, J. (1995) Starvation stress modulates the expression of the Aspergillus nidulans brlA regulatory gene. Microbiology 141, 21–28.

    CAS  Article  Google Scholar 

Download references

Acknowledgements

This project was supported financially by the Hungarian National Office for Research and Technology (grant reference number OMFB 01501/2006), the GENOMNANOTECH-DEBRET (RET-06/2004) and by the SOLE-MEAT Ltd., Szolnok, Hungary.

Author information

Affiliations

Authors

Corresponding author

Correspondence to T. Emri.

Rights and permissions

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

Reprints and Permissions

About this article

Cite this article

Tóth, V., Antal, K., Gyémánt, G. et al. Optimization of Coprogen Production in Neurospora crassa. BIOLOGIA FUTURA 60, 321–328 (2009). https://doi.org/10.1556/ABiol.60.2009.3.9

Download citation

Keywords

  • Siderophore
  • coprogen
  • Neurospora crassa
  • response surface methodology
  • L-ornithine-N5-monooxygenase