A substrate-multiplexed platform for profiling enzymatic potential of plant family 1 glycosyltransferases
Abstract Plants have expanded various biosynthetic enzyme families to produce a wide diversity of natural products; however, most enzymes encoded in plant genomes remain uncharacterized, highlighting the need for new functional genomic approaches. Here, we report a platform enabling the rapid functi...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-07-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-61530-6 |
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| author | Sasilada Sirirungruang Vincent Blay Elys P. Rodriguez Yasmine F. Scott Khanh M. Vuu Collin R. Barnum Paul H. Opgenorth Fanzhou Kong Yuanyue Li Oliver Fiehn Patrick M. Shih |
| author_facet | Sasilada Sirirungruang Vincent Blay Elys P. Rodriguez Yasmine F. Scott Khanh M. Vuu Collin R. Barnum Paul H. Opgenorth Fanzhou Kong Yuanyue Li Oliver Fiehn Patrick M. Shih |
| author_sort | Sasilada Sirirungruang |
| collection | DOAJ |
| description | Abstract Plants have expanded various biosynthetic enzyme families to produce a wide diversity of natural products; however, most enzymes encoded in plant genomes remain uncharacterized, highlighting the need for new functional genomic approaches. Here, we report a platform enabling the rapid functional characterization of plant family 1 glycosyltransferases, which serve important roles in plant development, defense, and communication. Using substrate-multiplexed reactions, mass spectrometry, and automated analysis, we screen 85 enzymes against a diverse library of 453 natural products, for a total of nearly 40,000 possible reactions. The resulting dataset reveals a widespread promiscuity and a strong preference for planar, hydroxylated aromatic substrates among family 1 glycosyltransferases. We also characterize glycosyltransferases with an unusually wide substrate scope and with a non-canonical Cys-Asp catalytic dyad. This work establishes a widely-applicable enzymatic screening pipeline, reflects the immense glycosylation capability of plants, and has implications in biocatalysis, metabolic engineering, and gene discovery. |
| format | Article |
| id | doaj-art-1187bf58fd5e40fa90c452a416cc0d45 |
| institution | DOAJ |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-1187bf58fd5e40fa90c452a416cc0d452025-08-20T03:05:10ZengNature PortfolioNature Communications2041-17232025-07-0116111210.1038/s41467-025-61530-6A substrate-multiplexed platform for profiling enzymatic potential of plant family 1 glycosyltransferasesSasilada Sirirungruang0Vincent Blay1Elys P. Rodriguez2Yasmine F. Scott3Khanh M. Vuu4Collin R. Barnum5Paul H. Opgenorth6Fanzhou Kong7Yuanyue Li8Oliver Fiehn9Patrick M. Shih10Department of Plant and Microbial Biology, University of CaliforniaJoint BioEnergy InstituteDepartment of Chemistry, University of CaliforniaJoint BioEnergy InstituteJoint BioEnergy InstituteDepartment of Plant Biology, University of CaliforniaJoint BioEnergy InstituteDepartment of Chemistry, University of CaliforniaWest Coast Metabolomics Center, University of CaliforniaDepartment of Chemistry, University of CaliforniaDepartment of Plant and Microbial Biology, University of CaliforniaAbstract Plants have expanded various biosynthetic enzyme families to produce a wide diversity of natural products; however, most enzymes encoded in plant genomes remain uncharacterized, highlighting the need for new functional genomic approaches. Here, we report a platform enabling the rapid functional characterization of plant family 1 glycosyltransferases, which serve important roles in plant development, defense, and communication. Using substrate-multiplexed reactions, mass spectrometry, and automated analysis, we screen 85 enzymes against a diverse library of 453 natural products, for a total of nearly 40,000 possible reactions. The resulting dataset reveals a widespread promiscuity and a strong preference for planar, hydroxylated aromatic substrates among family 1 glycosyltransferases. We also characterize glycosyltransferases with an unusually wide substrate scope and with a non-canonical Cys-Asp catalytic dyad. This work establishes a widely-applicable enzymatic screening pipeline, reflects the immense glycosylation capability of plants, and has implications in biocatalysis, metabolic engineering, and gene discovery.https://doi.org/10.1038/s41467-025-61530-6 |
| spellingShingle | Sasilada Sirirungruang Vincent Blay Elys P. Rodriguez Yasmine F. Scott Khanh M. Vuu Collin R. Barnum Paul H. Opgenorth Fanzhou Kong Yuanyue Li Oliver Fiehn Patrick M. Shih A substrate-multiplexed platform for profiling enzymatic potential of plant family 1 glycosyltransferases Nature Communications |
| title | A substrate-multiplexed platform for profiling enzymatic potential of plant family 1 glycosyltransferases |
| title_full | A substrate-multiplexed platform for profiling enzymatic potential of plant family 1 glycosyltransferases |
| title_fullStr | A substrate-multiplexed platform for profiling enzymatic potential of plant family 1 glycosyltransferases |
| title_full_unstemmed | A substrate-multiplexed platform for profiling enzymatic potential of plant family 1 glycosyltransferases |
| title_short | A substrate-multiplexed platform for profiling enzymatic potential of plant family 1 glycosyltransferases |
| title_sort | substrate multiplexed platform for profiling enzymatic potential of plant family 1 glycosyltransferases |
| url | https://doi.org/10.1038/s41467-025-61530-6 |
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