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Literature summary for 5.5.1.12 extracted from

  • Sugai, Y.; Ueno, Y.; Hayashi, K.; Oogami, S.; Toyomasu, T.; Matsumoto, S.; Natsume, M.; Nozaki, H.; Kawaide, H.
    Enzymatic (13)C labeling and multidimensional NMR analysis of miltiradiene synthesized by bifunctional diterpene cyclase in Selaginella moellendorffii (2011), J. Biol. Chem., 286, 42840-42847.
    View publication on PubMedView publication on EuropePMC

Protein Variants

Protein Variants Comment Organism
D391G/D392G site-directed mutagenesis of DIDD motif residues, the mutant lacks type-B activity Selaginella moellendorffii
D611G/D612G site-directed mutagenesis of DDLMD motif residues, the mutant lacks type-A cyclase activity Selaginella moellendorffii

Organism

Organism UniProt Comment Textmining
Selaginella moellendorffii G9MAN7
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Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
geranylgeranyl diphosphate BDTC-like enzyme, miltiradiene synthase converts geranylgeranyl diphosphate to a diterpene hydrocarbon product with a molecular mass of 272 Da. Mutation in the type-B active motif of BDTC-like enzyme miltiradiene synthase, D611G/D612G, abolishes the cyclase activity, whereas (+)-copalyl diphosphate, the reaction intermediate from the conversion of geranylgeranyl diphosphate to the hydrocarbon product, rescues the cyclase activity of the mutant to form a diterpene hydrocarbon. Another mutant lacking type-A activity, D391G/D392G, accumulates copalyl diphosphate as the reaction intermediate Selaginella moellendorffii (+)-copalyl diphosphate
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?
additional information miltiradiene synthesis reaction with geranylgeranyl diphosphate to miltiradiene, EC 4.2.3.131, via (+)-copalyl diphosphate intermediate, not ent-copalyl diphosphate. The enzyme catalyzes the successive two-step type-B (protonation-initiated cyclization) and type-A (ionization-initiated cyclization) reactions of geranylgeranyl diphosphate. The two aspartate-rich motifs, 389DIDD and 611DDLMD, that are required for the type-B and type-A cyclization reactions, respectively. Product analysis by GC-MS and NMR spectroscopy Selaginella moellendorffii ?
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Synonyms

Synonyms Comment Organism
BDTC
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Selaginella moellendorffii
bifunctional diterpene cyclase
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Selaginella moellendorffii

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
28
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assay at Selaginella moellendorffii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
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assay at Selaginella moellendorffii

General Information

General Information Comment Organism
metabolism bifunctional diterpene cyclases are involved in hormone and defense compound biosyntheses in bryophytes and gymnosperms, respectively Selaginella moellendorffii
additional information enzyme structure by two-dimensional and three-dimensional NMR Selaginella moellendorffii