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Functional Characterization of ent-Copalyl Diphosphate Synthase and Kaurene Synthase Genes from Coffea arabica L

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Author(s):
Ivamoto-Suzuki, Suzana Tiemi ; Celedon, Jose Miguel ; Yuen, Macaire M. S. ; Kitzberger, Ci ; Silva Domingues, Douglas ; Bohlmann, Jorg ; Protasio Pereira, Luiz Filipe
Total Authors: 7
Document type: Journal article
Source: Journal of Agricultural and Food Chemistry; v. 71, n. 42, p. 11-pg., 2023-10-10.
Abstract

The biochemical profile of coffee beans translates directly into quality traits, nutraceutical and health promoting properties of the coffee beverage. Ent-kaurene is the ubiquitous precursor for gibberellin biosynthesis in plants, but it also serves as an intermediate in specialized (i.e., secondary) diterpenoid metabolism that leads to a diversity of more than 1,000 different metabolites. Nutraceutical effects on human health attributed to diterpenes include antioxidant, anticarcinogenic, and anti-inflammatory properties. Cafestol (CAF) and kahweol (KAH) are two diterpenes found exclusively in the Coffea genus. Our objective was to identify and functionally characterize genes involved in the central step of ent-kaurene production. We identified 17 putative terpene synthase genes in the transcriptome of Coffea arabica. Two ent-copalyl diphosphate synthase (CaCPS) and three kaurene synthase (CaKS) were selected and manually annotated. Transcript expression profiles of CaCPS1 and CaKS3 best matched the CAF and KAH metabolite profiles in different tissues. CaCPS1 and CaKS3 proteins were heterologously expressed and functionally characterized. CaCPS1 catalyzes the cyclization of geranylgeranyl diphosphate (GGPP) to ent-copalyl diphosphate (ent-CPP), which is converted to ent-kaurene by CaKS3. Knowledge about the central steps of diterpene formation in coffee provides a foundation for future characterization of the subsequent enzymes involved in CAF and KAH biosynthesis. (AU)

FAPESP's process: 17/01455-2 - Biochemical and molecular basis of the metabolic network of diterpenes, bioactive and natural compounds in coffee
Grantee:Suzana Tiemi Ivamoto
Support Opportunities: Scholarships in Brazil - Post-Doctoral
FAPESP's process: 16/10896-0 - Can hexanoic acid signaling modulate the transcriptome, metabolome and pathogen development in coffee trees?
Grantee:Douglas Silva Domingues
Support Opportunities: Research Grants - Young Investigators Grants