Functional metagenomic screening identifies an unexpected β-glucuronidase
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Authors
Neun, Stefanie
Brear, Paul
Campbell, E
Tryfona, Theodora
El Omari, Kamel
Wagner, Armin
Dupree, Paul
Hyvonen, Marko
Journal Title
Nature Chemical Biology
ISSN
1552-4450
Publisher
Nature Research
Type
Article
This Version
AM
Metadata
Show full item recordCitation
Hollfelder, F., Neun, S., Brear, P., Campbell, E., Tryfona, T., El Omari, K., Wagner, A., et al. Functional metagenomic screening identifies an unexpected β-glucuronidase. Nature Chemical Biology https://doi.org/10.17863/CAM.84202
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 (GH) 3 family member. This GH 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 characterised it as a promiscuous catalyst and structural analysis suggests regions of divergence from homologous GH3 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)
Embargo Lift Date
2025-05-05
Identifiers
This record's DOI: https://doi.org/10.17863/CAM.84202
This record's URL: https://www.repository.cam.ac.uk/handle/1810/336784
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