ent-Kaurene is a key intermediate in the biosynthesis of the plant hormone gibberellin. In ent-kaurene biosynthesis in flowering plants, two diterpene cyclases (DTCs), ent-copalyl diphosphate (ent-CDP) synthase (ent-CPS) and ent-kaurene synthase (KS), catalyse the cyclization of geranylgeranyl diphosphate to ent-CDP and ent-CDP to ent-kaurene, respectively. In contrast, the moss Physcomitrella patens has a bifunctional ent-CPS/KS (PpCPS/KS) that catalyses both cyclization reactions. To gain more insight into the functional diversity of ent-kaurene biosynthetic enzymes in land plants, we focused on DTCs in the lycophyte Selaginella moellendorffii. The present paper describes the characterization of two S. moellendorffii DTCs (SmKS and SmDTC3) in vitro. SmDTC3 converted ent-CDP into ent-16α-hydroxykaurane and also used other CDP stereoisomers as substrate. Remarkably, SmKS, which produces ent-kaurene from ent-CDP, showed similar substrate selectivity: both SmKS and SmDTC3 synthesized sandaracopimaradiene from normal CDP. Therefore, the diversity of substrate recognition among KSs from other plants was investigated. PpCPS/KS could use normal CDP and syn-CDP as well as ent-CDP as substrate. In contrast, lettuce KS showed high specificity for ent-CDP, and rice KS recognized only ent-CDP. Our studies imply that ancient KS having low substrate specificity has evolved to be specific for ent-CDP to the biosynthesis of gibberellin.
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Gibberellin-biosynthetic ent-kaurene synthases in higher plants do not require their non-catalytic domains for the catalysis.
Biochem J
June 2024
Faculty of Agriculture, Yamagata University, Tsuruoka, Yamagata 997-8555, Japan.
ent-Kaurene is a biosynthetic intermediate diterpene of phytohormone gibberellins, and is biosynthesized from geranylgeranyl diphosphate via ent-copalyl diphosphate (ent-CDP). The successive cyclization is catalyzed by two distinct diterpene synthases, ent-CDP synthase (ent-CPS) and ent-kaurene synthase (KS). Homologs of these diterpene synthase genes have been reported to be involved in the biosynthesis of specialized-metabolic diterpenoids for defense in several plant species, including rice (Oryza sativa).
View Article and Find Full Text PDFCharacterization of diterpene synthase genes in Brachypodium distachyon, a monocotyledonous model plant, provides evolutionary insight into their multiple homologs in cereals.
Biosci Biotechnol Biochem
December 2023
Faculty of Agriculture, Yamagata University, Tsuruoka, Yamagata, Japan.
Article Synopsis
- * In crops, variations in the genes for these enzymes may have arisen from duplication and adaptation for specialized functions, while the model plant Brachypodium distachyon has a simpler gene set related to gibberellin biosynthesis, including one ent-CPS gene and four KS-like genes.
- * Detailed studies revealed that BdCPS and BdKS1 in B. distachyon are crucial for gibberellin production, whereas BdKSL2 and BdKSL3 likely
Functional kaurene-synthase-like diterpene synthases lacking a gamma domain are widely present in Oryza and related species.
Biosci Biotechnol Biochem
August 2021
Faculty of Agriculture, Yamagata University, Tsuruoka, Yamagata, Japan.
Various diterpene synthases have been functionally identified in cultivated rice (Oryza sativa). These are the homologs of ent-copalyl diphosphate (ent-CDP) synthase and ent-kaurene synthase (KS) that are responsible for the biosynthesis of gibberellins, diterpenoid phytohormones. We isolated a cDNA encoding full-length OsKSL12, a previously uncharacterized KS like (KSL) enzyme that consists of a β-domain and an α-domain with an active center, but lacks an N-terminal γ-domain.
View Article and Find Full Text PDFCharacterization of diterpene synthase genes in the wild rice species Oryza brachyatha provides evolutionary insight into rice phytoalexin biosynthesis.
Biochem Biophys Res Commun
September 2018
Biotechnology Research Center, The University of Tokyo, Bunkyo-ku, 113-8657, Japan.
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- Cultivated rice (Oryza sativa) produces various diterpenoids, including gibberellins (GAs) and phytoalexins crucial for plant defense, formed from geranylgeranyl diphosphate through specific enzymatic pathways.
- Comparisons between cultivated rice and its wild relative, Oryza brachyantha, reveal differences in their diterpene synthase genes, particularly in the types of cyclase enzymes that influence phytoalexin production.
- The study suggests that the acquisition of a specific enzyme variant (syn-CPS) in cultivated rice was a significant evolutionary development that enabled the production of oryzalexin S, a key phytoalexin for rice defense.
The rice ent-KAURENE SYNTHASE LIKE 2 encodes a functional ent-beyerene synthase.
Biochem Biophys Res Commun
May 2015
Hokkaido Agricultural Research Center, National Agriculture and Food Research Organization (NARO), Toyohira-ku, Sapporo 062-8555, Japan; Graduate School of Agriculture, Hokkaido University, Kita-ku, Sapporo 060-8589, Japan. Electronic address:
The rice genome contains a family of kaurene synthase-like (OsKSL) genes that are responsible for the biosynthesis of various diterpenoids, including gibberellins and phytoalexins. While many OsKSL genes have been functionally characterized, the functionality of OsKSL2 is still unclear and it has been proposed to be a pseudogene. Here, we found that OsKSL2 is drastically induced in roots by methyl jasmonate treatment and we successfully isolated a full-length cDNA for OsKSL2.
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