I want to do an assay to see, if an ordered enzyme is active or not. It's a glucosyltransferase from S. mutans which hydrolyses sucrose into fructose and glucose. It then connects the glucose-monomers to dextran. Our idea is to detect the free fructose. There are many tests for this, for example the Fehling-Test or the Tollen-Test. The trouble is, that they have to be heated on a waterbath - I think that's very suitable for microwell-plates. Maybe someone here has a better idea how to measure the enzyme's activity?

  • $\begingroup$ If you've access to a PCR machine or a heating plate with a microplate with a heating block you can use that $\endgroup$ – Michael_A Aug 9 '17 at 6:58
  • $\begingroup$ There are indirect enzymatic assays to detect fructose based on NADH production by glycolytic enzymes. $\endgroup$ – canadianer Aug 9 '17 at 17:04
  • $\begingroup$ Yes, true, but they are very expensive (about 400 Euro). Thanks for the answers, but we now sticked in to a new method with 3,5-dinitrosalicylic acid (photometrically), which also detects free fructose. $\endgroup$ – Simon Pflug Aug 10 '17 at 8:18
  • $\begingroup$ What about this kit? sigmaaldrich.com/catalog/product/sigma/fa20 $\endgroup$ – alec_djinn Aug 10 '17 at 9:39

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