S. (Suzanne) Wolterink-van Loo: “Biosynthesis using aldolases”

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6 Mar 2009 13:30
Unit: Wageningen University
Location: Aula, building 362, Gen. Foulkesweg 1, Wageningen
Organisation: Wageningen University
Promotor: prof.dr. J. van Oost (Microbiology)
Promotor: Prof.dr. W.M de Vos

 An ideal enzyme for biotechnological application has a long shelf life (stability), acts on cheap substrates and is highly stereospecific. Aldolases are enzymes, which can synthesize carbon-carbon bonds, generating a new stereogenic centre. In this thesis, research is described about the discovery, production and engineering of such aldolases for the production of nitrogen-heterocycles in biosynthetic routes. One enzyme, dihydrodipicolinate synthase from the bacterium Thermoanaerobacter tengcongensis appeared very specific for (S)-ASA, and was not really suitable for application purposes. 2-keto-3-deoxygluconate aldolase form the crenarchaeon Sulfolobus acidocaldarius appeared to be a very stable enzyme, with activity on a range of different substrates. To be able to use this enzyme in biocascade reactions with mesophilic enzymes, it was randomly optimized to increase activity at low-temperature, leading to a single optimized mutant, with increased activity between 35 and 60°C. Using a computational design approach we aimed for the stereospecific synthesis of the stereo-isomer 5-azido-4(S)-hydroxy-2-oxopentanoic acid. 
Title thesis: “Characterization and engineering of thermophilic aldolases. Synthesizing Nitrogen-heterocycles in biosynthetic routes”
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