Date of Award

2012

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Biology

First Advisor

White, Catherine D. Dr.

Abstract

Organophosphates (OP) are a class of pesticide widely used in agriculture and are found in a number of insecticides and chemical warfare agents. OPs are extremely dangerous in that they inhibit cholinesterase function, which disrupts proper activity of the nervous system. . OPs are also harmful due to their broad target range and toxicity for nontarget species (Horne, Qiu, Ollis, Russell, & Oakeshott, 2006) . Furthermore, following the introduction and wide spread use of pesticides, great environmental concerns have arisen. Some of the methods used for removing OP residues are impractical and costly, or are themselves environmentally hazardous (Horne et al., 2006). Despite these concerns, pesticides have been considered valuablel tools in agriculture given that they kill potential disease causing organisms, control insects, weeds, and other pests, and help reduce crop loss. Recently, the potential use of environmentally safe enzymes for the decontamination of pesticides and chemical warfare agents has become the focus of many studies. Therefore, the objective of this study was to clone, functionally express, and examine the catalytic activity of the organophosphagte degrading enzyme organophosphorus hydrolase (OPH). A 1,098 bp DNA fragment containing opd, the gene encoding OPH, was successfully amplified by Polymerase Chain Reaction from Flavobacterium sp. Opd was then cloned into an expression vector and inserted into E. coli by electrophoration. Expression of OPH in the E. coli was verified using a colony blot SDS-PAGE. OPH was isolated and a functional assay to demonstrate the catalytic activity of the protein revealed that the protein successfully degraded the substrate paraoxon.

Download

Share

COinS