You are here:
POSSIBLE ERRORS IN ASSAY FOR B-GLYCOSIDASE ACTIVITY
Citation:
Chadwick, R., S. George, J. Allison, AND D. Talley. POSSIBLE ERRORS IN ASSAY FOR B-GLYCOSIDASE ACTIVITY. U.S. Environmental Protection Agency, Washington, D.C., EPA/600/J-95/150.
Description:
Because intestinal B-glycosidase enzymes activate the toxicity and/or carcinogenicity of many environmental chemicals, accurate analysis of their activities is toxicologically important. owever, previous work in this lab indicated that widespread use of the glycosides of p-nitrophenol, as assay substrates, may have yielded low estimates of B-glycosidase activities. ata suggested that, in the presence of nitroreductase(e), the hydrolysis product, D-nitrophenol would be reduced to 2-aminophenol, which would not be detected in the assay. his study was designed to investigate the influence of nitroreductase and O2 on the determination of B-glycosidase activities by spectroscopic and gas chromatographic analyses The results demonstrated that in the presence of intestinal nitroreductase(s) or O2, B-glycosidase activities were significantly reduced. naerobic incubation and addition of excess 3,4-dichloronitrobenzene (DCNB), a substrate for nitroreductase, significantly increased the apparent activity of the principle glycosidase enzymes, B-glucosidase (BG1), B-glucuronidase (BGr), and B-galactosidase (BGa) in cecal incubation mixtures from adult male Fischer 344 rate.