The glycosyltransferase library
Activity screening uses either recombinant GTs purified from Escherichia coli, or protein extracts of the bacterial cells. These are incubated with scaffolds and sugar donors of interest. Reaction mixes are routinely analyzed using HPLC, MS and NMR to confirm product identity and the region/enantio-selectivity of catalytic activity. A predictive capacity has been established using detailed analyses of substrate recognition, catalytic activity and gene phylogeny. This know-how enables rapid selection of GT clades for screening novel scaffolds and can by-pass the need to explore the entire library. |
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Glytech has capability to identify relevant GT sequences in genome databases, clone the genes and prepare recombinant GT enzymes for biochemical characterisation. The current Glytech GT library contains 107 recombinant enzymes of arabidopsis and 33 recombinant enzymes of rice as well as engineered variants of native enzymes. Within the Glytech database, an additional 120 rice sequences and 177 grape sequences have been annotated ready for cloning and expansion of the library.