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 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.

Flavonoids are a class of plant secondary metabolites that fulfill many functions in planta and also benefit human health. Glucosylation of flavonoids, catalyzed by glucosyltransferases (GTs), help flavonoids perform their important roles. Citrus paradisi (grapefruit) is known to be active in flavonoid metabolism and possesses a variety of secondary product GTs. This research is designed to test the hypothesis that PGT10 is a flavonoid glucosyltransferase. PGT10 was cloned, expressed, purified, and tested for GT activity with a variety of phenolic substrates. Efforts to identify the reaction catalyzed by PGT10 continue. A second project was designed to test the hypothesis that multiple bands obtained by RT-PCR of PGT 5/6 found in root cDNA represented a set of homologous genes. Hypothesis has been confirmed by the homologous fragments found. Additional structural information is presented.