Functional metagenomic screening in microfluidic droplets identifies a β-glucuronidase in an unprecedented sequence neighbourhood
Neun, S.; Brear, P.; Campbell, E.; Tryfona, T.; El Omari, K.; Wagner, A.; Dupree, P.; Hyvonen, M.; Hollfelder, F.
Show 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 {beta}-glucuronidase activity. We identified SN243, a genuine {beta}-glucuronidase with little homology to previously studied enzymes of this type, as a glycoside hydrolase (GH) 3 family member. This GH family had no recorded evidence of {beta}-glucuronidases at the outset of this study, 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.
Matching journals
The top 3 journals account for 50% of the predicted probability mass.
Similar papers in this journal
Similar papers in this journal
- Functional metagenomics reveals an alternative, broad-specificity pathway for metabolism of carbohydrates in human gut commensal bacteria 97%
- Broadly applicable and accurate protein design by integrating structure prediction networks and diffusion generative models 96%
- Structural insights into the mechanism of the sodium/iodide symporter (NIS) 96%
Similar papers in this journal
- Evolution of dynamical networks enhances catalysis in a designer enzyme 96%
- Global analysis of biosynthetic gene clusters reveals conserved and unique natural products in entomopathogenic nematode-symbiotic bacteria 95%
- Label-free composition determination for biomolecular condensates with an arbitrarily large number of components 95%
"Similar papers" are the closest papers from that journal in the model's embedding space. They show what the match is built on, but the ranking comes mostly from a classifier over the whole training set, not from these examples alone.