Skip to main content
SpringerLink
Log in

Production of drug metabolites by immobilised Cunninghamella elegans: from screening to scale up

  • Short Communication
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Cunninghamella elegans is a fungus that has been used extensively as a microbial model of mammalian drug metabolism, whilst its potential as a biocatalyst for the preparative production of human drug metabolites has been often proposed, little effort has been made to enable this. Here, we describe a workflow for the application of C. elegans for the production of drug metabolites, starting from well-plate screening assays leading to the preparative production of drug metabolites using fungus immobilised either in alginate or as a biofilm. Using 12- and 96-well plates, the simultaneous screening of several drug biotransformations was achieved. To scale up the biotransformation, both modes of immobilisation enabled semi-continuous production of hydroxylated drug metabolites through repeated addition of drug and rejuvenation of the fungus. It was possible to improve the productivity in the biofilm culture for the production of 4′-hydroxydiclofenac from 1 mg/l h to over 4 mg/l h by reducing the incubation time for biotransformation and the number of rejuvenation steps.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1

References

  1. Amadio J, Casey E, Murphy C (2013) Filamentous fungal biofilm for production of human drug metabolites. Appl Microbiol Biotechnol 97:5955–5963

    Article  CAS  PubMed  Google Scholar 

  2. Amadio J, Gordon K, Murphy CD (2010) Biotransformation of flurbiprofen by Cunninghamella species. Appl Environ Microbiol 76:6299–6303

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  3. Amadio J, Murphy CD (2011) Production of human metabolites of the anti-cancer drug flutamide via biotransformation in Cunninghamella species. Biotechnol Lett 33:321–326

    Article  CAS  PubMed  Google Scholar 

  4. Asha S, Vidyavathi M (2009) Cunninghamella—a microbial model for drug metabolism studies—a review. Biotechnol Adv 27:16–29

    Article  CAS  PubMed  Google Scholar 

  5. Bucke C (1987) Cell immobilization in calcium alginate. Method Enzymol 135:175–189

    Article  CAS  Google Scholar 

  6. Corcoran O, Lindon JC, Hall R, Ismail IM, Nicholson JK (2001) The potential of F-19 NMR spectroscopy for rapid screening of cell cultures for models of mammalian drug metabolism. Analyst 126:2103–2106

    Article  CAS  PubMed  Google Scholar 

  7. Davis PJ, Campos J, Muthukumar V (1992) Hydroxylation of ibuprofen by Cunninghamella elegans. In: Abstr Pap Am Chem Soc 203: 125-MEDI

  8. Dlugonski J, Paraszkiewicz K, Sedlaczek L (1997) Maintenance of steroid 11-hydroxylation activity in immobilized Cunninghamella elegans protoplasts. World J Microbiol Biotechnol 13:469–473

    Article  CAS  Google Scholar 

  9. Dragan CA, Peters FT, Bour P, Schwaninger AE, Schaan SM, Neunzig I, Widjaja M, Zapp J, Kraemer T, Maurer HH, Bureik M (2011) Convenient gram-scale metabolite synthesis by engineered fission yeast strains expressing functional human P450 systems. Appl Biochem Biotechnol 163:965–980

    Article  CAS  PubMed  Google Scholar 

  10. Griffiths DA, Best DJ, Jezequel SG (1991) The screening of selected microorganisms for use as models of mammalian drug-metabolism. Appl Microbiol Biotechnol 35:373–381

    CAS  PubMed  Google Scholar 

  11. Guengerich FP (2009) Introduction: human metabolites in safety testing (MIST) issue. Chem Res Toxicol 22:237–238

    Article  CAS  PubMed  Google Scholar 

  12. Julsing MK, Cornelissen S, Buhler B, Schmid A (2008) Heme-iron oxygenases: powerful industrial biocatalysts? Curr Opin Chem Biol 12:177–186

    Article  CAS  PubMed  Google Scholar 

  13. Kang S, Kang SY, Lee EJ, Lim YH, Liu KH, Hur HG (2007) 4 ‘-Hydroxydiclofenac is the major metabolite of diclofenac by Cunninghamella elegans ATCC 9245 and Mucor ramannianus r-56. Drug Metab Rev 39:356

    Google Scholar 

  14. Maguire T, Novik E, Schloss R, Yarmush M (2006) Alginate-PLL microencapsulation: Effect on the differentiation of embryonic stem cells into hepatocytes. Biotechnol Bioengin 93:581–591

    Article  CAS  Google Scholar 

  15. Osorio-Lozada A, Surapaneni S, Skiles GL, Subramanian R (2008) Biosynthesis of drug metabolites using microbes in hollow fiber cartridge reactors: Case study of diclofenac metabolism by Actinoplanes species. Drug Metab Dispos 36:234–240

    Article  CAS  PubMed  Google Scholar 

  16. Peart PC, Chen ARM, Reynolds WF, Reese PB (2012) Entrapment of mycelial fragments in calcium alginate: a general technique for the use of immobilized filamentous fungi in biocatalysis. Steroids 77:85–90

    Article  CAS  PubMed  Google Scholar 

  17. Shanmugam B, Luckman S, Summers M, Bernan V, Greenstein M (2003) New approaches to augment fungal biotransformation. J Indust Microbiol Biotechnol 30:308–314

    Article  CAS  Google Scholar 

  18. Sutherland JB, Freeman JP, Heinze TM, Moody JD, Parshikov IA, Williams AJ, Zhang D (2001) Oxidation of phenothiazine and phenoxazine by Cunninghamella elegans. Xenobiotica 31:799–809

    Article  CAS  PubMed  Google Scholar 

  19. Webster R, Pacey M, Winchester T, Johnson P, Jezequel S (1998) Microbial oxidative metabolism of diclofenac: production of 4’-hydroxydiclofenac using Epicoccum nigrum IMI354292. Appl Microbiol Biotechnol 49:371–376

    Article  CAS  PubMed  Google Scholar 

  20. Williams AJ, Deck J, Freeman JP, Chiarelli MP, Adjei MD, Heinze TM, Sutherland JB (2007) Transformation of flumequine by the fungus Cunninghamella elegans. Chemosphere 67:240–243

    Article  CAS  PubMed  Google Scholar 

  21. Winn M, Foulkes JM, Perni S, Simmons MJH, Overton TW, Goss RJM (2012) Biofilms and their engineered counterparts: A new generation of immobilised biocatalysts. Catal Sci Technol 2:1544–1547

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We acknowledge financial assistance from the Enterprise Ireland Commercialisation Fund (CF/2011/1618) and the Environmental Protection Agency (2012-WRM-PhD-5).

Author information

Authors and Affiliations

  1. School of Biomolecular and Biomedical Science, University College Dublin, Belfield, Dublin 4, Ireland

    Laura Quinn, Rita Dempsey, Ayla Kane & Cormac D. Murphy

  2. School of Chemical and Bioprocess Engineering, University College Dublin, Belfield, Dublin 4, Ireland

    Eoin Casey

Authors
  1. Laura Quinn
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Rita Dempsey
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Eoin Casey
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ayla Kane
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Cormac D. Murphy
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Cormac D. Murphy.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 1432 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Quinn, L., Dempsey, R., Casey, E. et al. Production of drug metabolites by immobilised Cunninghamella elegans: from screening to scale up. J Ind Microbiol Biotechnol 42, 799–806 (2015). https://doi.org/10.1007/s10295-015-1594-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10295-015-1594-9

Keywords

Search

Navigation