Biomolecular Engineering for Peptide-Based Biomaterials

Belinda Hartmann (2010). Biomolecular Engineering for Peptide-Based Biomaterials PhD Thesis, School of Chemical Engineering, The University of Queensland.

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Author Belinda Hartmann
Thesis Title Biomolecular Engineering for Peptide-Based Biomaterials
School, Centre or Institute School of Chemical Engineering
Institution The University of Queensland
Publication date 2010-05
Thesis type PhD Thesis
Supervisor Professor Anton Middelberg
Dr Waltraud Kaar
Dr Linda Lua
Total pages 176
Total colour pages 27
Total black and white pages 149
Subjects 09 Engineering
Abstract/Summary Bioactive peptides that can self-assemble reversibly, in response to external stimuli, are at the core of a new breed of “smart” peptides that form a complex array of novel biomaterials with potential applications in fields including tissue engineering, nanoelectronics, functional surface coating and food production. The majority of future markets associated with peptide-based biomaterials are characterized by high volume and low cost economics. This has limited the deployment of these new biomaterials because the expensive, synthetic peptide production routes currently used by industry to produce commercial quantities of peptide therapeutics are not compatible with this cost dynamic. Consequently, this project aimed to contribute to ongoing efforts to achieve cost-effective, efficient and scalable means of producing peptide by utilizing recombinant DNA technology and microbial hosts. Three separate bioprocesses have been designed, investigated and reported in this thesis. The first process design is based on the insoluble expression of a peptide-concatamer fusion protein in a bacterial host, and represents an attempt at optimizing a conventional, commercially-available system to test its suitability as an industrial bioprocess. The second bioprocess utilizes a soluble expression system, also in a bacterial host, and exemplifies a substantial departure from traditional process designs for recombinant peptide production. A final methodology is described which employs a yeast expression host and relies on the incorporation of additional unit operations into an existing industrial bioprocess, to create a dual production route for a target peptide and a target protein. The key research findings in this body of work are related to the development of selective precipitation methods, which were employed throughout each of the described processes to various degrees. Simple organic solvents, salts and acids were used to exploit the property differences between peptides and proteins in order to selectively partition peptide from heterogenous mixtures. The selective precipitation methods devised successfully minimized the need for expensive and time-consuming chromatography processes which are ordinarily required to obtain high yields of pure peptide. In terms of recombinant peptide production, the Selective Solvent Precipitation (SSP) techniques developed as part of this PhD have genuine potential as an industrial replacement for expensive chromatographic unit operations that currently limit the cost-effectiveness of conventional production routes.
Keyword Peptide, biomaterials, recombinant, bacteria, yeast, precipitation, expression, cost, chromatography-free, bioprocess
Additional Notes 22-27, 34, 54, 74, 82, 86-91, 99, 100, 119, 124, 125, 138, 140, 145, 151, 154, 174

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Created: Tue, 12 Oct 2010, 15:15:16 EST by Miss Belinda Hartmann on behalf of Library - Information Access Service