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Structural And Functional Analysis Of Grapefruit Flavonol-Specific-3-O-Gt Mutant P145t, Sangam Kandel Mr
Structural And Functional Analysis Of Grapefruit Flavonol-Specific-3-O-Gt Mutant P145t, Sangam Kandel Mr
Electronic Theses and Dissertations
This research is focused on the study of the effect of mutating proline 145 to threonine on the substrate and regiospecificity of flavonol specific 3-O-glucosyltransferase (Cp3GT). While the mutant P145T enzyme did not glucosylate anthocyanidins, it did glucosylate flavanones and flavones in addition to retaining activity with flavonols. HPLC was used for product identification and showed mutant P145T glucosylated naringenin at the 7-OH position forming naringenin-7-O-glucoside and flavonols at the 3-OH position. Homology modeling and docking was done to predict the acceptor substrate recognition pattern and models were validated by experimental results. In other related work, …
Cloning, Expression, And Biochemical Characterization Of Recombinant Putative Glucosyltransferases Clone 3 And 8 From Grapefruit (Citrus Paradisi), Deborah Hayford
Cloning, Expression, And Biochemical Characterization Of Recombinant Putative Glucosyltransferases Clone 3 And 8 From Grapefruit (Citrus Paradisi), Deborah Hayford
Electronic Theses and Dissertations
The grapefruit plant, Citrus paradisi, tends to accumulate high levels of flavonoid glycosides such as flavanones and flavones. Flavonoids have a vast array of important functions in plants and also in humans. Glucosyltransferases (GTs) are enzymes responsible for glucosylation reactions. In our pursuit to study the structure and function of flavonoid GTs, we have used molecular approaches to identify, clone, express, and functionally characterize the enzymes. This research was designed to test the hypothesis that PGT3 is a flavonoid glucosyltransferase and is subject to biochemical regulation. PGT3 has been tested for GT activity with compounds representing subclasses of flavonoids …
Cloning, Heterologous Expression In Yeast, And Biochemical Characterization Of Recombinant Putative Glucosyltransferase Clones 9 And 11 From Grapefruit (Citrus Paradisi), Anye Wamucho
Electronic Theses and Dissertations
Flavonoids are plant secondary metabolites that play diverse roles in plants and human health. These compounds in most part exist in the glucosylated form. Grapefruit accumulates high levels of glucosylated flavonoids. Plant secondary product glucosyltransferases (GTs) catalyze the glucosylation reaction, but due to low homology at both the nucleotide and amino acid sequence level of different GTs, it is not possible to ascribe function based on sequence only. The hypotheses that PGT clones 9 and 11 are plant secondary product GTs and are biochemically regulated were tested. PGT 9 has been cloned into Pichia pastoris using the pPICZA and pPICZAα …