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Title: Characterization of a secondary alcohol oxidase from Brevundimonas vesicularis
Author: Podtetenieff, John J.
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2006
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Enantiomerically pure chiral secondary alcohols are important intermediates in the fine chemical, pharmaceutical and agrochemical industries. Many processes exist for generating enantiomerically pure compounds. One of these processes, deracemisation, functions by the combination of repeated cycles of enantioselective enzymatic oxidation and non-specific chemical reduction to convert a racemic mixture to a single enantiomer. The aim of this project was to provide an effective secondary alcohol oxidase for the oxidation step of deracemisation processes. To this end, poly(vinyl alcohol) oxidase from Brevundimonas vesicularis was chosen due to its broad substrate specificity. A purification method was developed and assays carried out establishing its preference for R-secondary alcohols. Due to the slow growth of B. vesicularis, it was decided to clone the gene encoding PVA oxidase. The goal was to develop a robust biocatalysed for industrial applications; therefore, rapid and reliable access to large quantities of the enzyme was necessary. This could be provided by an over-expressed recombinant PVA oxidase. Attempts at cloning were made with Southern hybridisation, inverse PCR, and analysis of an expression library. Much forward progress was made, but the gene remained elusive. Subsequently, the use of crude PVA oxidase preparations was investigated. These have the potential to quickly and practically oxidise a variety of chiral secondary alcohols.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available