1887

Abstract

, a strictly anaerobic micro-organism and inhabitant of the human intestine, has been shown to produce the azoreductase enzyme AzoC, an NAD(P)H-dependent flavin oxidoreductase. This enzyme reduces azo dyes to aromatic amines, which are carcinogenic in nature. A significant amount of work has been completed that focuses on the activity of this enzyme; however, few studies have been completed that focus on the physiology of azo dye reduction. Dye reduction studies coupled with growth studies in the presence of ten different azo dyes and in media of varying complexities were completed to compare the growth rates and dye-reducing activity of WT cells, a Δ knockout, and , a non-azoreductase-producing control bacterium. The presence of azo dyes significantly increased the generation time of in rich medium, an effect that was not seen in minimal medium. In addition, azo dye reduction studies with the Δ knockout suggested the presence of additional functional azoreductases in this medically important bacterium. Overall, this study addresses a major gap in the literature by providing the first look, to our knowledge, at the complex physiology of upon azo dye exposure and the effect that both azo dyes and the azoreductase enzyme have on growth.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.000212
2016-02-01
2020-09-26
Loading full text...

Full text loading...

/deliver/fulltext/micro/162/2/330.html?itemId=/content/journal/micro/10.1099/mic.0.000212&mimeType=html&fmt=ahah

References

  1. Ahmad N., Drew W. L., Plorde J. J.. 2010; Sherris Medical Microbiology, 5th edn. New York: McGraw Hill;
    [Google Scholar]
  2. Bradford M. M.. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem72:248–254 [CrossRef][PubMed]
    [Google Scholar]
  3. Brown M. A., Devito S. C.. 1993; Predicting azo dye toxicity. Crit Rev Environ Sci Technol23:249–324 [CrossRef]
    [Google Scholar]
  4. Chen H.. 2006; Recent advances in azo dye degrading enzyme research. Curr Protein Pept Sci7:101–111 [CrossRef][PubMed]
    [Google Scholar]
  5. Chen H., Wang R. F., Cerniglia C. E.. 2004; Molecular cloning, overexpression, purification, and characterization of an aerobic FMN-dependent azoreductase from Enterococcus faecalis. Protein Expr Purif34:302–310 [CrossRef][PubMed]
    [Google Scholar]
  6. Chung K. T., Cerniglia C. E.. 1992; Mutagenicity of azo dyes: structure–activity relationships. Mutat Res277:201–220 [CrossRef][PubMed]
    [Google Scholar]
  7. Encinas-Yocupicio A. A., Razo-Flores E., Sánchez-Diaz F., dos Santos A. B., Field J. A., Cervantes F. J.. 2006; Catalytic effects of different redox mediators on the reductive decolorization of azo dyes. Water Sci Technol54:165–170 [CrossRef][PubMed]
    [Google Scholar]
  8. Fuchs A. R., Bonde G. J.. 1957; The nutritional requirements of Clostridium perfringens. J Gen Microbiol16:317–329 [CrossRef][PubMed]
    [Google Scholar]
  9. Levine W. G.. 1991; Metabolism of azo dyes: implication for detoxication and activation. Drug Metab Rev23:253–309 [CrossRef][PubMed]
    [Google Scholar]
  10. Morrison J. M., John G. H.. 2013; The non-enzymatic reduction of azo dyes by flavin and nicotinamide cofactors under varying conditions. Anaerobe23:87–96 [CrossRef][PubMed]
    [Google Scholar]
  11. Morrison J. M., John G. H.. 2015; Non-classical azoreductase secretion in Clostridium perfringens in response to sulfonated azo dye exposure. Anaerobe34:34–43 [CrossRef][PubMed]
    [Google Scholar]
  12. Morrison J. M., Wright C. M., John G. H.. 2012; Identification, isolation and characterization of a novel azoreductase from Clostridium perfringens. Anaerobe18:229–234 [CrossRef][PubMed]
    [Google Scholar]
  13. Morrison J., Dai S., Ren J., Taylor A., Wilkerson M., John G., Xie A.. 2014; Structure and stability of an azoreductase with an FAD cofactor from the strict anaerobe Clostridium perfringens. Protein Pept Lett21:523–534 [CrossRef][PubMed]
    [Google Scholar]
  14. Nakanishi M., Yatome C., Ishida N., Kitade Y.. 2001; Putative ACP phosphodiesterase gene (acpD) encodes an azoreductase. J Biol Chem276:46394–46399 [CrossRef][PubMed]
    [Google Scholar]
  15. Punj S., John G. H.. 2008; Physiological characterization of Enterococcus faecalis during azoreductase activity. Microb Ecol Health Dis20:65–73 [CrossRef]
    [Google Scholar]
  16. Rafii F., Coleman T.. 1999; Cloning and expression in Escherichia coli of an azoreductase gene from Clostridium perfringens and comparison with azoreductase genes from other bacteria. J Basic Microbiol39:29–35 [CrossRef][PubMed]
    [Google Scholar]
  17. Stolz A.. 2001; Basic and applied aspects in the microbial degradation of azo dyes. Appl Microbiol Biotechnol56:69–80 [CrossRef][PubMed]
    [Google Scholar]
  18. van der Zee F. P., Lettinga G., Field J. A.. 2000; The role of (auto)catalysis in the mechanism of an anaerobic azo reduction. Water Sci Technol42:301–308
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.000212
Loading
/content/journal/micro/10.1099/mic.0.000212
Loading

Data & Media loading...

Supplements

Supplementary Data

PDF
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