1.14.11.13: gibberellin 2beta-dioxygenase

This is an abbreviated version!
For detailed information about gibberellin 2beta-dioxygenase, go to the full flat file.

Word Map on EC 1.14.11.13

Reaction

gibberellin 1
+
2-oxoglutarate
+
O2
=
2beta-hydroxygibberellin 1
+
succinate
+
CO2

Synonyms

2-oxoglutarate-dependent dioxygenase of gibberellin biosynthesis, AaGA2ox1, AaGA2ox2, AaGA2ox3, AaGA2ox4, AtGA2ox2, AtGA2ox8, C19-GA 2-oxidase, C20 GA2ox, C20-gibberellin deactivation enzyme, GA 2-ox, GA 2-oxidase, GA 2-oxidase 6, GA 2-oxidase 7, GA 2-oxidase A1, GA 2-oxidase1, GA 2-oxidase3, GA 2beta,3beta-hydroxylase, GA 2ODD, GA2ox, GA2ox1, GA2ox2, GA2ox3, GA2ox4, GA2ox5, GA2ox6, GA2ox7, gibberellin 2-oxidase, gibberellin 2-oxidase 1, gibberellin 2-oxidase 2, gibberellin 2-oxidase 3, gibberellin 2-oxidase 4, gibberellin 2-oxidase 6, gibberellin 2beta-hydroxylase, giberellin 2-oxidase 2, MdGA2ox1, PsGA2ox1, PsGA2ox2, PvGA2ox5b, PvGA2ox9a

ECTree

     1 Oxidoreductases
         1.14 Acting on paired donors, with incorporation or reduction of molecular oxygen
             1.14.11 With 2-oxoglutarate as one donor, and incorporation of one atom of oxygen into each donor
                1.14.11.13 gibberellin 2beta-dioxygenase

Purification

Purification on EC 1.14.11.13 - gibberellin 2beta-dioxygenase

Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
24 h after application of 14C-labelled substrate to the shoot apex, whole shoots are harvested, rinsed with water, same genotype shoots combined and homogenized in 80% methanol water, stirring overnight at 4°C, filtration, residue re-extracted in methanol for 1 h, filtered, combined filtrates vacuum dried, weak acid fraction prepared by solvent partitioning, anion exchange, and C18 SPE (no solvent partitioning for substrate gibberellin 9), purified samples are analyzed by reverse-phase HPLC with online radiomonitoring, identification of radiolabeled products by GC-MS
Escherichia coli cells with recombinant enzyme grown at 37°C, harvested and suspended in lysis buffer (100 mM Tris-HCl, pH 7.5), sonication in ice water, centrifuged, supernatant collected and stored at -80°C for enzyme activity assays
-
Escherichia coli with recombinant enzyme are centrifuged, resuspended in a BugBuster Protein Extraction Reagent, centrifuged, supernatants eluted through a GST-Bind resin with 50 mM Tris buffer, pH 8.0, containing 10 mM reduced glutathione, storage at -80°C
-
glutathione agarose bead affinity chromatography and gel filtration
glutathione Sepharose 4B column chromatography
-
recombinant enzyme
Sephadex G-25 column chromatography and Superdex G-25 column chromatography, Mini S column chromatography and gel filtration chromatography
-
two gibberellin 2beta-hydroxylases partially purified
-