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Structure to function studies in UDP-glucose dehydrogenases and nitroreductases
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The thesis is divided into two parts corresponding to structural studies on two
different proteins. The first part concerns the study of two UDP-glucose
dehydrogenases (UGDs) from Sphingomonas elodea ATCC 31461 and
Burkholderia cepacia IST 408, both involved in exopolysaccharide production.
Their relevance arises because some of these bacterial exopolysaccharides are
valuable as established biotechnological products, the former case, whilst others
are highly problematic, when used by pathogens in biofilm formation over
biological surfaces, as the latter case, namely in the human lungs. The goal of
these studies is to increase our knowledge regarding UGDs structural
properties, which can potentiate either the design of activity enhancers to
respond to the increased demand of useful biofilms, or the design of inhibitors
of biofilm production, in order to fight invading pathogens present in several
infections. The thesis reports the production and crystallisation of both proteins,
the determination of initial phases by single-wavelength anomalous dispersion
(SAD) in S. elodea crystals using a seleno-methionine isoform, and phasing of
B. cepacia crystals by molecular replacement (MR) using the S. elodea model,
as well as the refinement, structural analysis and comparison between the
several UGDs structures available during this work.(...)
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Universidade Nova de Lisboa. Instituto de Tecnologia Química e Biológica