Date of Graduation

8-2016

Document Type

Thesis

Degree Name

Master of Science in Microelectronics-Photonics (MS)

Degree Level

Graduate

Department

Microelectronics-Photonics

Advisor/Mentor

Robert Beitle

Committee Member

Jin-Woo Kim

Second Committee Member

Ralph Henry

Third Committee Member

Rick Wise

Keywords

Applied sciences, Carbon nanotubes, Cell wall permeability, Escherichia coli, Membrane damage

Abstract

This research investigated the use of carbon nanotubes (CNTs) as a treatment to increase the permeability of a bacterial cell wall. Recombinant Escherichia coli BL21 (DE3) containing a plasmid that expressed Green Fluorescent Protein (GFP) and -lactamase were exposed to CNTs under various levels of agitation for different times. Fluorescence assay for GFP, optical absorbance for -lactamase activity, and Transmission Electron Microscopy (TEM) were used to determine the amount of released protein, and visually examine the permeability enhancement of the cells, respectively. It was found that more -lactamase was present in the culture fluid after treatment with CNTs in a dose dependent manner. Indeed, CNTs can lyse the cells up to 90% of maximum when compared to lysozyme treatment. Based on TEM, it is believed that this treatment damaged the cell walls to make E. coli permeable, causing periplasm proteins and enzymes to leak out into the medium. Consequently, CNTs can be used as lysis agents when it is undesirable to add an additional enzyme (lysozyme) to cause the release of intracellular proteins.

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