APPLICATION OF 6-NITROCOUMARIN AS A SUBSTRATE FOR THE FLUORESCENT DETECTION OF NITROREDUCTASE ACTIVITY IN Sporothrix schenckii

Authors

  • Cheila Denise Ottonelli Stopiglia Universidade Federal do Pampa
  • Mariana Carissimi Municipal Department of Environmental Protection
  • Tatiane Caroline Daboit Universidade Federal do Rio Grande do Sul
  • Valter Stefani Universidade Federal do Rio Grande do Sul; Department of Organic Chemistry
  • Valeriano Antonio Corbellini Universidade de Santa Cruz do Sul; Department of Chemistry and Physics
  • Maria Lucia Scroferneker Universidade Federal do Rio Grande do Sul; Institute of Basic Health Sciences; Department of Microbiology

Abstract

Introduction Sporothrix schenckii is a thermal dimorphic pathogenic fungus causing a subcutaneous mycosis, sporotrichosis. Nitrocoumarin represents a fluorogenic substrate class where the microbial nitroreductase activity produces several derivatives, already used in several other enzyme assays. The objective of this study was the analysis of 6-nitrocoumarin (6-NC) as a substrate to study the nitroreductase activity in Sporothrix schenckii. Methods Thirty-five samples of S. schenckii were cultivated for seven, 14 and 21 days at 35 °C in a microculture containing 6-nitrocoumarin or 6-aminocoumarin (6-AC) dissolved in dimethyl sulfoxide or dimethyl sulfoxide as a negative control, for posterior examination under an epifluorescence microscope. The organic layer of the seven, 14 and 21-day cultures was analyzed by means of direct illumination with 365 nm UV light and by means of elution on G silica gel plate with hexane:ethyl acetate 1:4 unveiled with UV light. Results All of the strains showed the presence of 6-AC (yellow fluorescence) and 6-hydroxylaminocoumarin (blue fluorescence) in thin layer chromatography, which explains the green fluorescence observed in the fungus structure. Conclusion The nitroreductase activity is widely distributed in the S. schenckii complex and 6-NC is a fluorogenic substrate of easy access and applicability for the nitroreductase activity detection.

Downloads

Download data is not yet available.

Downloads

Published

2013-09-01

How to Cite

Stopiglia, C. D. O., Carissimi, M., Daboit, T. C., Stefani, V., Corbellini, V. A., & Scroferneker, M. L. (2013). APPLICATION OF 6-NITROCOUMARIN AS A SUBSTRATE FOR THE FLUORESCENT DETECTION OF NITROREDUCTASE ACTIVITY IN Sporothrix schenckii . Revista Do Instituto De Medicina Tropical De São Paulo, 55(5), 353-356. Retrieved from http://www.revistas.usp.br/rimtsp/article/view/78670

Issue

Section

Mycology