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Presentation
Mutagenesis of a Flavonol- 3-O-Glucosyltransferase and the Effect on Enzyme Function
53rd Annual Meeting of the Phytochemical Society of North America
  • Lisa Carter, East Tennessee State University
  • Devaiah P. Shivakumar, East Tennessee State University
  • Cecelia A. McIntosh, East Tennessee State University
Document Type
Presentation
Publication Date
8-9-2013
Date Range
08/09/2014-08/13/2014
Description

Flavonoids are an important group of secondary metabolites found in plants and have a wide variety of properties. Some play a role in fl ower pigmentation, while others have antimicrobial properties. Glucosylation is an important modifi cation of fl avonoids and is mediated by glucosyltransferases. In this process, the enzyme transfers glucose from UDP-glucose to a specifi c position on the fl avonoid. Previous study from the lab characterized a glucosyltransferase from C. paradisi that is fl avonol specifi c. In this study an attempt has been made to study the structure and function of this fl avonol specifi c glucosyltransferase using site directed mutagenesis. The glutamine residue at position 87 of the Cp-3-O-GT enzyme was changed to isoleucine, the analogous residue in the 3-O-glucosyltransferase of Clitoria ternatea. Similarly, the histidine at position 154 was changed to tyrosine. We hypothesize that these mutations will change substrate specifi city. The glutamate at position 88 was changed to an aspartic acid. We hypothesize that this will change the regiospecifi city of the enzyme, as aspartic acid is the analogous residue found in some 7-O-glucosyltransferases. Finally, we introduced a double mutation with glutamine 87 becoming isoleucine and glutamate 88 becoming aspartic acid, with the hypothesis that both regiospecifi city and substrate specifi city will be changed.

Location
Raleigh, NC
Citation Information
Lisa Carter, Devaiah P. Shivakumar and Cecelia A. McIntosh. "Mutagenesis of a Flavonol- 3-O-Glucosyltransferase and the Effect on Enzyme Function" 53rd Annual Meeting of the Phytochemical Society of North America (2013)
Available at: http://works.bepress.com/cecilia-mcintosh/52/