Functional metagenomic screening identifies an unexpected β-glucuronidase.
Nat Chem Biol
Springer Science and Business Media LLC
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Neun, S., Brear, P., Campbell, E., Tryfona, T., El Omari, K., Wagner, A., Dupree, P., et al. (2022). Functional metagenomic screening identifies an unexpected β-glucuronidase.. Nat Chem Biol https://doi.org/10.1038/s41589-022-01071-x
The abundance of recorded protein sequence data stands in contrast to the small number of experimentally verified functional annotation. Here we screened a million-membered metagenomic library at ultrahigh throughput in microfluidic droplets for β-glucuronidase activity. We identified SN243, a genuine β-glucuronidase with little homology to previously studied enzymes of this type, as a glycoside hydrolase 3 family member. This glycoside hydrolase family contains only one recently added β-glucuronidase, showing that a functional metagenomic approach can shed light on assignments that are currently 'unpredictable' by bioinformatics. Kinetic analyses of SN243 characterized it as a promiscuous catalyst and structural analysis suggests regions of divergence from homologous glycoside hydrolase 3 members creating a wide-open active site. With a screening throughput of >107 library members per day, picolitre-volume microfluidic droplets enable functional assignments that complement current enzyme database dictionaries and provide bridgeheads for the annotation of unexplored sequence space.
BBSRC (BB/W000504/1, BB/T003545/1) and EPSRC (EP/H046593/1)
Engineering and Physical Sciences Research Council (EP/H046593/1)
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External DOI: https://doi.org/10.1038/s41589-022-01071-x
This record's URL: https://www.repository.cam.ac.uk/handle/1810/336784
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