Date of Award

2013

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Chemistry

Subject(s)

Microbiological chemistry, Microbial enzymes, Drug resistance in microorganisms, Antibiotics

Abstract

The critical need for new antibiotics and antibiotic targets is of great urgency to our society in order to combat drug resistance and ward off pathogenic bacteria. One new antibacterial target is the essential bacterial enzyme Peptidyl-tRNA Hydrolase I (Pth1). In the course of protein elongation, ribosomes stall and release peptidyl-tRNA. Peptidyl-tRNA hydrolase 1 releases the tRNA from the peptidyl-tRNA, allowing the tRNA to be recycled in the cell. Without Pthl hydrolysis, peptidyl-tRNAs will accumulate, slowing down and eventually stopping protein synthesis, leading to cell death. The fact that Pthl is an essential gene in E. coli suggests that its homologue in S. aureus and B. cereus are possible molecular therapeutic targets for the discovery of novel antibacterial agents. The overall goal of this project was to clone,express, and provide purified S. aureus and B. cereus Pthl protein for activity and inhibition screening. The study of this enzyme's characteristics, activity, inhibition, structure, and mechanism could reveal how the inhibition of Pthl affects the cell as a whole.

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