The ancient CYP716 family is a major contributor to the diversification of eudicot triterpenoid biosynthesis.
Triterpenoids are widespread bioactive plant defence compounds with potential use as pharmaceuticals, pesticides and other high-value products. Enzymes belonging to the cytochrome P450 family have an essential role in creating the immense structural diversity of triterpenoids across the plant kingdom. However, for many triterpenoid oxidation reactions, the corresponding enzyme remains unknown. Here we characterize CYP716 enzymes from different medicinal plant species by heterologous expression in engineered yeasts and report ten hitherto unreported triterpenoid oxidation activities, including a cyclization reaction, leading to a triterpenoid lactone. Kingdom-wide phylogenetic analysis of over 400 CYP716s from over 200 plant species reveals details of their evolution and suggests that in eudicots the CYP716s evolved specifically towards triterpenoid biosynthesis. Our findings underscore the great potential of CYP716s as a source for generating triterpenoid structural diversity and expand the toolbox available for synthetic biology programmes for sustainable production of bioactive plant triterpenoids.
Department of Plant Systems Biology, VIB, Technologiepark 927, B-9052 Ghent, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium.
Karel
K
Miettinen
Department of Plant Systems Biology, VIB, Technologiepark 927, B-9052 Ghent, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium.
Jacob
0000-0002-1134-9238
J
Pollier
Department of Organic Chemistry, Ghent University, B-9000 Ghent, Belgium.
Dieter
D
Buyst
Department of Plant Systems Biology, VIB, Technologiepark 927, B-9052 Ghent, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium.|Laboratory for Protein Biochemistry and Biomolecular Engineering, Department of Biochemistry and Microbiology, Ghent University, B-9000 Ghent, Belgium.|VIB Medical Biotechnology Center, B-9000 Ghent, Belgium.
Philipp
P
Arendt
Department of Organic Chemistry, Martin-Luther-University Halle-Wittenberg, D-06120 Halle (Saale), Germany.
René
R
Csuk
Department of Organic Chemistry, Martin-Luther-University Halle-Wittenberg, D-06120 Halle (Saale), Germany.
Sven
S
Sommerwerk
Department of Plant Systems Biology, VIB, Technologiepark 927, B-9052 Ghent, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium.
Tessa
0000-0001-9366-4727
T
Moses
Department of Plant Systems Biology, VIB, Technologiepark 927, B-9052 Ghent, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium.
Jan
J
Mertens
Department of Plant and Environmental Sciences, Weizmann Institute of Science, Rehovot 76100, Israel.
Prashant D
PD
Sonawane
Department of Plant Systems Biology, VIB, Technologiepark 927, B-9052 Ghent, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium.
Laurens
0000-0002-0221-9052
L
Pauwels
Department of Plant and Environmental Sciences, Weizmann Institute of Science, Rehovot 76100, Israel.
Asaph
A
Aharoni
Department of Organic Chemistry, Ghent University, B-9000 Ghent, Belgium.
José
J
Martins
Department of Microbiology, Immunology and Biochemistry, University of Tennessee Health Science Center, Memphis, Tennessee 38163, USA.
David R
DR
Nelson
Department of Plant Systems Biology, VIB, Technologiepark 927, B-9052 Ghent, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium.
Alain
0000-0002-1599-551X
A
Goossens
D044822:Biodiversity; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D019143:Evolution, Molecular; D010802:Phylogeny; D018521:Plant Physiological Phenomena; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D014315:Triterpenes / Q000378:metabolism*
D010940:Plant Proteins; D014315:Triterpenes; D003577:Cytochrome P-450 Enzyme System