Functional and Structural Insights into a Novel Promiscuous Ketoreductase of the Lugdunomycin Biosynthetic Pathway.
- Xiao X Molecular Biotechnology, Leiden University, PO Box 9505, 2300RA Leiden, The Netherlands.
- Elsayed SS Molecular Biotechnology, Leiden University, PO Box 9505, 2300RA Leiden, The Netherlands.
- Wu C State Key Laboratory of Microbial Technology, Institute of Microbial Technology, Shandong University, Qingdao, Shandong 266237, P. R. China.
- van der Heul HU Molecular Biotechnology, Leiden University, PO Box 9505, 2300RA Leiden, The Netherlands.
- Metsä-Ketelä M Department of Biochemistry and Food Chemistry, University of Turku, FIN-20014 Turku, Finland.
- Du C Molecular Biotechnology, Leiden University, PO Box 9505, 2300RA Leiden, The Netherlands.
- Prota AE Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, CH-5232 Villigen, Switzerland.
- Chen CC State Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Sciences, Hubei University, Wuhan, Hubei 43420, P. R. China.
- Liu W State Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Sciences, Hubei University, Wuhan, Hubei 43420, P. R. China.
- Guo RT State Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Sciences, Hubei University, Wuhan, Hubei 43420, P. R. China.
- Abrahams JP Molecular Biotechnology, Leiden University, PO Box 9505, 2300RA Leiden, The Netherlands.
- van Wezel GP Molecular Biotechnology, Leiden University, PO Box 9505, 2300RA Leiden, The Netherlands.
- 2020-08-26
Published in:
- ACS chemical biology. - 2020
English
Angucyclines are a structurally diverse class of actinobacterial natural products defined by their varied polycyclic ring systems, which display a wide range of biological activities. We recently discovered lugdunomycin (1), a highly rearranged polyketide antibiotic derived from the angucycline backbone that is synthesized via several yet unexplained enzymatic reactions. Here, we show via in vivo, in vitro, and structural analysis that the promiscuous reductase LugOII catalyzes both a C6 and an unprecedented C1 ketoreduction. This then sets the stage for the subsequent C-ring cleavage that is key to the rearranged scaffolds of 1. The 1.1 Å structures of LugOII in complex with either ligand 8-O-Methylrabelomycin (4) or 8-O-Methyltetrangomycin (5) and of apoenzyme were resolved, which revealed a canonical Rossman fold and a remarkable conformational change during substrate capture and release. Mutational analysis uncovered key residues for substrate access, position, and catalysis as well as specific determinants that control its dual functionality. The insights obtained in this work hold promise for the discovery and engineering of other promiscuous reductases that may be harnessed for the generation of novel biocatalysts for chemoenzymatic applications.
- Language
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- English
- Open access status
- hybrid
- Identifiers
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- DOI 10.1021/acschembio.0c00564
- PMID 32840360
- Persistent URL
- https://sonar.ch/global/documents/207015
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