Functional metagenomic screening identifies an unexpected β-glucuronidase.
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Authors
Campbell, Eleanor
Publication Date
2022-10Journal Title
Nat Chem Biol
ISSN
1552-4450
Publisher
Springer Science and Business Media LLC
Type
Article
This Version
AM
Metadata
Show full item recordCitation
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
Abstract
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.
Sponsorship
BBSRC (BB/W000504/1, BB/T003545/1) and EPSRC (EP/H046593/1)
Funder references
BBSRC (BB/T003545/1)
BBSRC (BB/W000504/1)
Engineering and Physical Sciences Research Council (EP/H046593/1)
Identifiers
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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