Author

Qi Ge

Advisor

Emerson, Joseph P.

Committee Member

Lewis, Edwin A.

Committee Member

Mlsna, Todd

Date of Degree

1-1-2012

Document Type

Graduate Thesis - Open Access

College

College of Arts and Sciences

Department

Department of Chemistry

Abstract

PrnB is a heme-containing enzyme, which catalyzes the ring rearrangement reaction of 7-chlorotryptophan to produce 3-(3-Chloro-2-nitrophenyl)pyrrole. This thesis describes the initial isolation and characterization of PrnB, the second enzyme associated with the pyrrolnitrin biosynthetic pathway in Burkholderia ambifaria. Additionally, alternative peroxidase reactivity was used to study how amino-acids bind to the substrate binding pocket of PrnB. The peroxidase activity of PrnB was measured using three different peroxidase activity assays at various pH values. The peroxidase data was compared to similar studies with the classic peroxidase, Horseradish peroxidase (HRP). Generally, PrnB showed weak peroxidase reactivity. However this weak reactivity was an experimental handhold, where tryptophan and other substrate binding events can be explored using classic inhibition steady-state kinetics. The rate of 2-aminophenol oxidation by PrnB was used as a model assay to monitor how molecules such as L-tryptophan, L-alanine, indole, L-phenylalanine, and L-tyrosine interact with the PrnB active site.

URI

https://hdl.handle.net/11668/20108

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