A New Exploration of Extracting and Purifying the Coenzyme F420 from Natural Sludge

A New Exploration of Extracting and Purifying the Coenzyme F420 from Natural Sludge

Loading document ...
Loading page ...


Author(s): ZHU Xiao-wen, LI Rong-gui

Download Full PDF Read Complete Article

DOI: 10.18483/ijSci.1213 294 639 62-67 Volume 6 - Mar 2017


Coenzyme F420 is one kind of special flavin cofactors which exist in some Archaea and bacteria, it is a low potential electron transfer carrier in methanogens. In this study, we designed three specific primers of F420-dependent glucose-6-phos-phate dehydrogenase gene to determine which environment contains the more abundant coenzyme F420, and methods for separating, extracting and purifying the coenzyme F420 from this environmental samples were carried out, the pure coenzyme F420 was obtained by some optimizing methods from the natural sludge , and the purity and concentration of coenzyme F420 we got is just as good as the FMN standard sample which confirmed by their fluorescence emission spectrum. These optimizing methods would save more energy and time to get the good quality coenzyme F420.


Coenzyme F420, Separation and Purification, Fluorescence Spectra


  1. Delafontaine. Biotechnol. 1979,1:71-74
  2. Eberhard Warkentin, Bjorn Mamat,et.al. ,Structures of F420H2:NADP+ oxidoreductase with and without its substrates bound. EMBO Journal 2001,6561-6569
  3. Retno S. Sudibyo, Umar A.Jenie. , Biomimetic experiment of enoyl-reduction process by F420-dependent enzyme obtained from Saccharopolyspora erythraea and the biosynthetic implication. Indonesian Journal of Biotechnology,1997,133-139
  4. Luke Bown, Mead S. Altowairish, et.al., Production of the Streptomyces scabies coronafacoyl phytotoxins involves a novel biosynthetic pathway with an F420-dependent oxidoreductase and a short-chain dehydrogenase/reductase, Molecular Microbiology. 2016, 00-00l
  5. B. Nocek, E. Evdokimova, M. Proudfoot, Structure of an Amide Bond Forming F420:γ-glutamyl Ligase from Archaeoglobus Fulgidus-A Member of a New Family of Nonribosomal Peptide Synthases, J Mol Biol. 2007, 456–469
  6. Cuong Quang Le, Ebenezer Joseph, oan Nguyen, Optimization of Expression and Purification of Recombinant Archeoglobus fulgidus F420H2:NADP+ Oxidoreductase, an F420 Cofactor Dependent Enzyme. Protein J , 2015,34:391-397
  7. Jeremy D ,Selengut, Daniel H. Haft. Unexpected Abundance of Coenzyme F420 -Dependent Enzymes in Mycobacterium tuberculosis and Other Actinobacteria, Journal of bacteriology, 2010, 5788–5798
  8. Gorren ACF, Mayer B, Tetrahydrobiopterin in nitric oxide synthesis: a novel biological role for pteridines. Curr. Drug Metab 2002;3:133–157
  9. Begley TP, Cofactor biosynthesis: an organic chemist's treasure trove. Nature Prod. Rep 2006;23:15–25
  10. Mercy A. Oyugi, Ghader Bashiri, Investigating the Reaction Mechanism of F420 -Dependent Glucose-6-phosphate Dehydrogenase from Mycobacterium tuberculosis: Kinetic Analysis of the Wild-Type and Mutant Enzymes, Biochemistry, 2016, 55, 5566−5577
  11. Ghader Bashiri, Christopher J. Squire, CrystalStructuresof F420-dependent Glucose-6-phosphateDehydrogenase FGD1 Involved in the Activation of the Anti-tuberculosis Drug Candidate PA-824 Reveal the Basis of Coenzyme and Substrate Binding ,The journal of biological chemistry, 2008,17531–17541
  12. Matthew Taylor, Colin Scott, Gideon Grogan, F420 -dependent enzymes – potential for applications in biotechnology.Trends in Biotechnology February 2013,63-65
  13. White RH, Biosynthesis of the methanogenic cofactors. Vitam. Horm 2001;61:299–337
  14. Delafontaine.M.J., H. P. Naveau, E.-J. Nyns, luorimetric monitoring of methanogenesis in anaerobic digestors. Biotechnol. Lett. 1979,1:71-74
  15. van Beelen, P., A. C. DiJkstra, G. D. Vogels, Quantitation of coenzyme F420 in methanogenic sludge by the use of reversed-phase high-performance liquid chromatography and a fluorescence detector. Eur. J. Microbiol. Biotechnol. 1983-18:67-69
  16. LEON G. GORRIS, THEO M. DE KOK, Relationship between Methanogenic Cofactor Content and Maximum Specific Methanogenic Activity of Anaerobic Granular Sludges Applied and environmental microbiology, 1988, 1126-1130
  17. Whitmore, T. N., S. P. Etheridge, D. A. Stafford, The evaluation of anaerobic digester performance by coenzyme F420 analysis. Biomass 1986-9:29-35
  18. Van Beelen, P., W. J. Geerts, A. Pol, Quantification of coenzymes and related compounds from methanogenic bacteria by high-performance liquid chromatography. Anal. Biochem. 1983, 131:285-290.
  19. Bashiri, G., C. J. Squire, N. J, Crystal structures of F420-dependent glucose-6-phosphate dehydrogenase FGD1 involved in the activation of the anti-tuberculosis drug candidate PA-824 reveal the basis of coenzyme and substrate binding. J. Biol. Chem. 2008, 283:17531–17541.
  20. Ney, Blair, Ahmed, F Hafna, The methanogenic redox cofactor F420 is widely synthesized by aerobic soil bacteria. The ISME journal,03 Mar 2017 11:125-137
  21. ENDANG PURWANTINI. Molecular Analysis of the Gene Encoding F420 -Dependent Glucose-6-Phosphate Dehydrogenase from Mycobacterium smegmatis. J OURNAL OF BACTERIOLOGY ,Apr. 1998, p. 2212–2219
  22. A. Elaaf Mohamed, F. Hafna Ahmed, Sundaram Arulmozhiraja, et. Protonation state of F 420 H 2 in the prodrug-activating deazaflavin dependent nitroreductase (Ddn) from Mycobacterium tuberculosis; Electronic Supplementary Material (ESI) for Molecular BioSystems. 2016
  23. P. CHEESEMAN, A. TOMS-WOOD, Isolation and Properties of a Fluorescent Compound, Factor420, from Methanobacterium Strain M.o.H. JOURNAL OF BACTERIOLOGY, 1972, 527-531

Cite this Article:

International Journal of Sciences is Open Access Journal.
This article is licensed under a Creative Commons Attribution 4.0 International (CC BY 4.0) License.
Author(s) retain the copyrights of this article, though, publication rights are with Alkhaer Publications.

Search Articles

Issue June 2023

Volume 12, June 2023

Table of Contents

World-wide Delivery is FREE

Share this Issue with Friends:

Submit your Paper