Date of Graduation

12-2018

Document Type

UAF Access Only - Thesis

Degree Name

Bachelor of Science in Chemical Engineering

Degree Level

Undergraduate

Department

Chemical Engineering

Advisor

Beitle, Robert

Abstract

The purpose of this work was to explore a different platform for antimicrobial peptide manufacturing. This alternative to conventional antibiotic synthesis offers certain advantages while overcoming drawbacks of the pharmaceutical manufacturing industry. The main goal was to prove that Pediococcus acidilacticicould be used as a recombinant-manufacturing platform. A peptide was to be introduced to P. acidilacticiand secrete an antifungal; followed by separation techniques to ultimately offer a naturally occurring alternative to synthetic antimicrobial drugs that are currently available. Initially, the path forward of this project was to insert a modified pSMB74plasmid into theP. acidilactici, grow the bacterium in MRS media, and separate the secreted antimicrobial product from the media by various techniques. However, due to multiple failed attempts to synthesize the modified plasmid, the project only progressed through the design phase of plasmid modification. There is still opportunity for this project to progress because in the end it was determined that pTRKH3-slpGFP and pTRKH2 shuttle vectors may secrete the desired product in both lactobacillus and Escherichia coli. Further opportunity for this project would start with inserting the desired expression machinery from the pSMB74 plasmid into the two shuttle vectors and continue onto the bio-reaction and separation techniques as discussed.

Keywords

pediococcus, plasmid, shuttle vector, recombinant manufacturing, peptide

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