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Direct detection of rutin-degrading isozymes with polyacrylamide gel electrophoresis

Authors
Journal
Analytical Biochemistry
0003-2697
Publisher
Elsevier
Publication Date
Volume
443
Issue
2
Identifiers
DOI: 10.1016/j.ab.2013.09.010
Keywords
  • Rutin-Degrading Enzyme
  • Isozyme
  • Page
  • Western Blotting
Disciplines
  • Biology
  • Physics

Abstract

Abstract Rutin-degrading enzymes (RDEs) specifically hydrolyze the glycosidic linkages of rutin, producing quercetin and rutinose. Here we report a reliable and sensitive polyacrylamide gel electrophoresis and staining method for the detection of RDE isozymes, which is based on the aqueous solubility difference between rutin and quercetin, as well as the ultraviolet absorbance of quercetin. With this novel method, we achieved a detection limit of 12ng with 107U of RDE activity, enabling us to detect at least five RDE isozymes in tartary buckwheat seeds.

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