Doctor of Philosophy (PhD)
Cyanobacteria show much promise in reducing biodegradable thermoplastic production costs; however, most currently characterized strains are ill-equipped to do so. The result of Objective I produced a high-throughput assay designed to discover existing cyanobacterial strains and rapidly characterize them as PHA-producers or potential PHA-producers. This assay will play an instrumental role in the attainment of a novel cyanobacteria environmental isolate capable of accumulating high levels of PHA naturally. Objective II produced an open source computer program which dramatically speeds the design of similar assays for any arbitrary genetic screening purpose. The program is not limited to this implementation alone. In fact, there are as many uses for this program as there are consensus and/or degenerate oligonucleotide probe applications. The project was released as open source in order to provide a means of constant growth and development by those who need it most. The case studies investigated during the preliminary research of Objective III provided key insights into the complex mechanisms involved in in vitro PHA synthase polymerization kinetics. Additionally, multiple hypothetical physical phenomena are proposed, as inferred from data from literature, which are capable of explaining the kinetic model behavior. All difficulties encountered during the course of Objective III, namely the recombinant protein expression and purification failures, are detailed so that the methods used may be avoided in future experiments. Even though Objective III was completed using an impure PHA synthase sample, it was still found conclusively that the conserved cyanobacteria-specific insertion of the model cyanobacterium PHA synthase is required for proper functionality. This conclusion is significant because it is evidence that the PHA synthase of cyanobacteria may possess a unique catalytic mechanism or method of interaction for multimerization.
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Lane, Courtney Edward, "Bioplastic Production in Cyanobacteria and Consensus Degenerate PCR Probe Design" (2015). LSU Doctoral Dissertations. 2777.