Cloning, Heterologous Expression in Yeast, and Biochemical Characterization of Recombinant Putative Glucosyltransferase Clones 9 and 11 from Grapefruit (Citrus paradisi)

<p>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 <em>Pichia pastoris</em> using the pPICZA and pPICZAα vectors, expressed, enriched, and screened for GT activity with a variety of phenolic substrates. Initial screens show catechol, gentisic acid, vanillin, and p-hydroxylphenylacetic acid as potential substrates for the PGT 9 protein. PGT 11 has been successfully cloned into pPICZA for transformation into yeast, expression, and subsequent characterization.</p>

Name thesis:degree_name

MS (Master of Science)

Level thesis:degree_level

Thesis - unrestricted

Discipline thesis:degree_discipline

Biology

Year dc:date.issued

2012

Author dc:creator

• Wamucho, Anye

• PSPG Box
• Glucosyltransferase
• Citrus paradisi
• Phenolics
• Flavonoids
• Biology
• Life Sciences

Statement dc:rights

• Copyright by the authors.

Repository record dc:identifier

https://dc.etsu.edu/etd/1214

OAI identifier oai:identifier

oai:dc.etsu.edu:etd-2400