Bibliography: leaves 235-250. / 297 leaves : ill. ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Aims to develop the process for Poly-[beta]-hydroxybutyrate recovery from recombinant E. coli, using homogenization and centrifugation as the basic unit operations, with effects directed to reduce the Poly-[beta]-hydroxybutyrate overall production cost. / Thesis (Ph.D.)--University of Adelaide, Dept. of Chemical Engineering, 1999

Thesis (Ph.D.)--University of Adelaide, Dept. of Chemical Engineering, 1999. / Bibliography: leaves 235-250.

Thesis (M. Phil.)--Hong Kong University of Science and Technology, 2003. / Includes bibliographical references. Also available in electronic version. Access restricted to campus users.

published_or_final_version / abstract / toc / Civil Engineering / Master / Master of Philosophy

Includes bibliographical references (leaves G-1-G-11 ). / vii, 1 v. (various pagings) : ill. (some col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Substatial developmental work is required to acheive economically attractive poly-hydroxyalkanoate (PHA) production. Aims to improve the understanding of PHA production by recombinant Escherichia coli, with a view to guiding future developmental work. Also suggests numerous areas where further improvements to PHB production are possible, and proposes ways in which they could be implemented. / Thesis (Ph.D.)--Adelaide University, Dept. of Chemical Engineering, 2001

The main objective of this thesis is to investigate the impact of microbial extracellular polymeric substances (EPS) on sediment stability and the related factors which influence “biogenic stabilisation” as a basis to the prediction of sediment erosion and transport. The ability to make direct and sensitive measurements of the physical properties of the biofilm is a critical demand to further understanding of the overall biostabilisation processes. Therefore, attention has been focused on developing a new technique, Magnetic Particle Induction (MagPI) for measuring the adhesive properties of the biofilm. MagPI determines the relative adhesive properties or “stickiness” of the test surface, whether a biofilm, a sediment or other submerged material. The technique may have future applications in physical, environmental and biomedical research. Newly developed Magnetic Particle Induction(MagPI) and traditional techniques Cohesive Strength Meter (CSM) for the determination of the adhesion/cohesion of the substratum were used to assess the biostabilisation capacity of aquatic microorganisms. Whilst these devices determine slightly different surface properties of the bed, they were found to complement each other, increasing the range of measurements that could be made and presented a strong correlation in the overlapping portion of the data. It is recognized that microorganisms inhabiting natural sediments significantly mediate the erosive response of the bed (“ecosystem engineers”) through the secretion of naturally adhesive organic material (EPS: extracellular polymeric substances). Interactions between main biofilm consortia microalgae, cyanobacteria and bacteria in terms of their individual contribution to the EPS pool and their relative functional contribution to substratum stabilisation were investigated. The overall stabilisation potential of the various assemblages was impressive, as compared to controls. The substratum stabilisation by estuarine microbial assemblages was due to the secreted EPS matrix, and both EPS quality (carbohydrates and proteins) and quantity (concentration) were important in determining stabilisation. Stabilisation was significantly higher for the bacterial assemblages than for axenic microalgal assemblages. The peak of engineering effect was significantly greater in the mixed assemblage as compared to the bacterial and axenic diatom culture. This work confirmed the important role of heterotrophic bacteria in “biostabilisation” and highlighted the interactions between autotrophic and heterotrophic biofilm components of the consortia. An additional approach, to investigate the impact of toxins on biostabilisation capacity of aquatic organism was performed on cultured bacterial and natural freshwater biofilm. The data suggest a different mode of triclosan (TCS) action ranging from suppressing metabolisms to bactericidal effects depending on the TCS concentration. The inhibitory effect of triclosanon bacterial and freshwater biofilms was confirmed. This information contributes to the conceptual understanding of the microbial sediment engineering that represents an important ecosystem function and service in aquatic habitats.

PHAs are a class of biopolymers consisting of (R)-3-hydroxy-fatty acids and are produced by the majority of eubacteria and some archaeal bacteria as carbon storage material. In general, PHA is synthesised when a carbon source is available in excess while another essential nutrient is limited. The key enzyme of PHA biosynthesis, the PHA synthase, catalyses the polymerisation of the substrate (R)-3-hydroxyacyl-CoA to PHA accompanied by the release of coenzyme A. PHA is stored intracellularly as inclusions, the so-called PHA granules. When the external carbon source becomes exhausted, bacteria can metabolise these carbon inclusions by degradation of the polymer. PHA granules are water-insoluble, spherical inclusions of approximately 50-500 nm in diameter which consist of a hydrophobic polyester core surrounded by a phospholipid layer with embedded and attached proteins. One could consider isolated PHA granules as bio-beads due to their structure and size. In this study we tested if the PHA synthase can be used as an anchor molecule in order to display proteins of interest at the PHA granule surface. Furthermore, these modified PHA granules were analysed for their potential applicability as bio-beads in biotechnological procedures. The concept of using the PHA synthase as granule-anchoring molecule for display of proteins of interest was established by the functional display of the ß- galactosidase at PHA granules. This “proof of concept” was followed by the display of biotechnologically more interesting proteins. The IgG binding domain of protein A as well as streptavidin, which is known for its biotin binding ability, were fused to the PHA synthase, respectively, and therefore localised at the PHA granule surfaces during PHA granule assembly, resulting in functional bio-protein A -beads and bio-streptavidin-beads. Moreover, their applicability in biotechnological assays was demonstrated. Recently, we fused the green fluorescent protein (GFP) to the PHA synthase and demonstrated that the PHA granule assembly does not start randomly distributed in the cytoplasm but occurred localised at or near the cell poles. To further investigate if the localisation of the PHA granule formation process is due to polar positional information inherent to the PHA synthase, different mutated versions of the PHA synthase of Cupriavidus necator were created and analysed for a potential alteration in localisation. Furthermore, the phasin protein PhaP1 of C. necator was fused to HcRed, a far-red fluorescent protein, and localisation studies were accomplished when the fusion protein was expressed under different conditions in Escherichia coli.