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Title: DNA nanostructures for biotechnological applications
Author: Sohrabi, Bouran
ISNI:       0000 0004 7660 3414
Awarding Body: UCL (University College London)
Current Institution: University College London (University of London)
Date of Award: 2019
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Deoxyribonucleic acid (DNA) is a versatile biomolecule which can be used for the rational design and assembly of nanoscale structures. This thesis explores the use of functional DNA- enzyme nanostructures for applications in biocatalysis and for directed motion on the nanoscale. In the first part of this thesis, a DNA scaffold was outlined for the display and immobilization of enzyme cascades. Confinement or spatial organization of enzyme cascades is adopted in biological systems to prevent loss of reactive intermediates and to facilitate substrate conversion in chemically complex and crowded intracellular environments. We adopt a strategy to create concentrated enzyme assemblies directed by a DNA structure generated by F29 rolling circle amplification (RCA). These DNA assemblies, DNA nanoballs, were investigated for the display of two bi-enzyme systems. Firstly, a horseradish peroxidase and glucose oxidase enzyme pair, and secondly, a transaminase and norcoclaurine synthase bi- enzyme system for the synthesis of biotechnologically relevant benzylisoquinoline (BIA) precursors. The second part of this thesis concerns the use of enzymatic catalysis as a means of affecting the motion of a nanoscale DNA structure. Molecular movement on the micro and nanoscales is a fundamental feature of biological systems, and recreating this functionality represents an important step in the realization of intelligent synthetic devices for directed transport and chemotaxis in response to stimuli. While directed motion has been shown for DNA structures on predefined tracks to which they are hybridized, enzymatic catalysis has not been investigated as an approach to controlling the motion of DNA nanostructures. We show that the motion of a DNA structure tethered to multiple lysine decarboxylase molecules is enhanced by its substrate, L-lysine the 'fuel', in a concentration dependent manner, based on nanoparticle tracking analysis (NTA) and DLS analyses.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available