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4.2.3.146: cyclooctat-9-en-7-ol synthase

This is an abbreviated version!
For detailed information about cyclooctat-9-en-7-ol synthase, go to the full flat file.

Word Map on EC 4.2.3.146

Reaction

geranylgeranyl diphosphate
+
H2O
=
cyclooctat-9-en-7-ol
+
diphosphate

Synonyms

cetB2, CotB2, CYC, diterpene cyclooctatin synthase

ECTree

     4 Lyases
         4.2 Carbon-oxygen lyases
             4.2.3 Acting on phosphates
                4.2.3.146 cyclooctat-9-en-7-ol synthase

Crystallization

Crystallization on EC 4.2.3.146 - cyclooctat-9-en-7-ol synthase

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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purified recombinant native and SeMet-labeled enzyme CotB2 with bound substrate analogue geranylgeranyl thiodiphosphate, hanging drop vapor diffusion method, mixing of 0.001 ml of 10 mg/ml protein, in 20 mM Tris-HCl, pH 8.0, 5 mM MgSO4, with or without 1 mM substrate, with 0.001 ml of reservoir solution, and equilibration against 0.5 ml of reservoir solution, 20°C, the reservoir solution contains 0.1 M HEPES-NaOH, pH 7.0-8.5, and 1.4-1.5 M ammonium formate for the apoenzyme, 0.1 M Tris-HCl, pH 8.0, and 2.4 M ammonium formate for the apo-SeMet-enzyme, and 0.1 M bicine-NaOH, pH 9.0, and 20% PEG 6000 for the substrate-bound enzyme, X-ray diffraction structure determination and analysis. Molecular replacement. Only a single Mg2+ ion is present in the observed structure, coordinated by the Asn220, Ser224, and Glu228 side chains belonging to the canonical NSE/DTE motif
sitting drop vapor diffusion method, using 0.8 M NaH2PO4/K2HPO4 pH 7.8
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