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Full-Text Articles in Life Sciences
Modification, Verification Of Sequence And Optimization Of Expression Of P297f An Inactive Mutant Of Flavonol Specific Glucosyltransferase From Grapefruit (Cp3gt), Sarah Fox
Undergraduate Honors Theses
Citrus fruits are widely consumed and can offer various health benefits. One enzyme found in grapefruits, Citrus paradisi flavonol specific 3-O-glucosyltransferase (CP3GT), catalyzes the addition of glucose to one specific flavonoid class and at only one site. These flavonoids are plant secondary metabolites that can be used in a variety of plant functions including signaling and protection. The only class of flavonoids that CP3GT glucosylates is flavonols, and this specificity is of interest to study for potential benefits in biotechnology and enzyme modeling. In order to study this enzyme and its structure, a variety of mutants were created using site-directed …
Preparation Of A Flavonol Specific Glucosyltransferase Found In Grapefruit And Site-Directed Mutants For Protein Crystallization, Aaron Birchfield
Preparation Of A Flavonol Specific Glucosyltransferase Found In Grapefruit And Site-Directed Mutants For Protein Crystallization, Aaron Birchfield
Electronic Theses and Dissertations
This research was designed to determine the conditions necessary to remove c-myc and 6x-His tags from a flavonol specific glucosyltransferase found in grapefruit (CP3GT) using thrombin in preparation for crystallization. X-ray crystallography of CP3GT crystals may elucidate structural features that account for flavonol specificity in some glucosyltransferase enzymes. A thrombin cleavage site was inserted into WT CP3GT and one mutant. Recombinant CP3GT was expressed in yeast and purified. Optimal conditions for thrombin digestion were explored. Digestion with 100U of thrombin for 2 hours at 4o C was optimal for removing tags from CP3GT. Storage at 4o C for …
Characterization Of Sip68 For Its Role In Plant Stress Signaling, Saroj Chandra Lohani
Characterization Of Sip68 For Its Role In Plant Stress Signaling, Saroj Chandra Lohani
Electronic Theses and Dissertations
Glucosyltransferases catalyze the transfer of glucose molecules from an active donor to acceptor molecules and are involved in many plant processes. SIP68, a tobacco glucosyltransferase protein, is a SABP2-interacting protein. It was identified in a yeast two-hybrid screen using SABP2 as bait and tobacco proteins as prey. SABP2, converts methyl salicylate to salicylic acid (SA) as a part of the signal transduction pathways in SA-mediated defense signaling. Subcellular localization is a crucial aspect of protein functional analysis to assess its biological function. The recombinant SIP68 tagged with eGFP was expressed transiently in Nicotiana benthamiana and observed under confocal microscopy. Fluorescent …
Characterization Of Sbip68: A Putative Tobacco Glucosyltransferase Protein And Its Role In Plant Defense Mechanisms, Abdulkareem O. Odesina
Characterization Of Sbip68: A Putative Tobacco Glucosyltransferase Protein And Its Role In Plant Defense Mechanisms, Abdulkareem O. Odesina
Electronic Theses and Dissertations
Plant secondary metabolites are essential for normal growth and development in plants ultimately affecting crop yield. They play roles ranging from appearance of the plants to defending against pathogen attack and herbivory. They have been used by humans for medicinal and recreational purposes amongst others. Glycosyltransferases catalyze the transfer of sugars from donor substrates to acceptors. Glucosyltransferases are a specific type of glycosyltransferases known to transfer glucose molecules from a glucose donor to a glucose acceptor (aglycone) producing the corresponding glucose secondary metabolite or glycone, in this case glucosides. It was hypothesized that SBIP68, a tobacco putative glucosyltransferase-like protein glucosylated …
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 …
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 …