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Jinping Du

Doctor of Philosophy, (Microbiology)
Study Completed: 2018
College of Sciences

Citation

Thesis Title
Design of Bacterial Polyester Beads for Recombinant Protein Production, Biomolecule Separation and Detection

Read article at Massey Research Online: MRO icon

Protein recovery and biomolecule detection is of great importance in biotech areas, but related process is expensive and complicated. In order to simplify relevant process, Mrs Du integrated self-cleavable tag technology or OBody technology with polyhydroxyalkanoate (PHA) bead display technology via a PhaC (PHA synthase) fusion strategy. By fusing PhaC to a target protein via a self-cleavable linker tag (either sortase or intein), the target protein could be produced on PHA beads and purified by specific cleavage, either by incubation with CaCl2 ± triglycine, or by a pH shift to 6. As such, a streamlined process was provided for production and purification of recombinant proteins. By fusing PhaC to differently customised OBody ligands, functional OBody beads could be easily obtained and directly used for lysozyme separation from a complex substrate, or for progesterone (P4) detection. Therefore, a streamlined process was provided for separation and detection of industrially important biomolecules.

Supervisors
Professor Bernd Rehm
Professor Rosie Bradshaw

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