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The use of biocatalysts for the synthesis of chiral compounds and fine chemicals is of significant interest to the chemical, pharmaceutical and industrial biotechnology sectors in the search for sustainable, cost effective, synthetic strategies. In a collaboration at UCL (with Prof J. M. Ward, Prof P. Dalby, and Prof G. Lye) we have been discovering new enzymes and using them in a range of applications. Enzymes include (i) Transketolases, including libraries of mutants that can accept a wide range of aliphatic aldehydes as well as aromatic substrates in high yields. (ii) Libraries of transaminases (>200) both wild-type and enzyme variants have been developed, together with assays. These have been used to convert many substrates in both single steps and enzyme cascades. (iii) Norcoclaurine synthases where in a collaboration with Prof J. M. Ward we have investigated the enzymes including their characterisation, substrate tolerance, and biocatalytic applications. We have also investigated the substrate-binding model that was subsequently validated by an X-ray crystal structure with Prof. N Keep (at Birkbeck). Remarkably we have recently discovered that these enzymes are also able to accept ketone substrates and other more challenging substrates. (iv) Methyl transferase enzymes to carry out the selective alkylation of a range of substrates. We have also been investigating the use of functional metagenomics to identify new enzymes (with Prof J. M. Ward and Prof Christine Orengo). Enzymes discovered using this approach include new transketolases, robust transaminases, epoxide hydrolases, ene-reductases, dehydrogenases and imine reductases. The in silico libraries generated from the sequenced metagenomes will provide a valuable source of enzymes in the future. In a collaboration with Prof T. Sheppard, Prof J. M. Ward and Prof G. Lye, bio-derived feedstocks have been used such as sugar beet waste derived biomass for conversion into higher value chemicals using enzymes. For example, a sugar beet pulp derived sugar readily reacted in transketolase reactions to give a rare heptulose and enzyme cascades for hydroxy-pyruvate supply were constructed. Also, the formation of amino-polyols from sugars was achieved using transaminases.
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