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2.5.1.54: 3-deoxy-7-phosphoheptulonate synthase

This is an abbreviated version!
For detailed information about 3-deoxy-7-phosphoheptulonate synthase, go to the full flat file.

Word Map on EC 2.5.1.54

Reaction

phosphoenolpyruvate
+
D-erythrose 4-phosphate
 +
H2O
=
3-deoxy-D-arabino-hept-2-ulosonate 7-phosphate
 +
phosphate

Synonyms

2-dehydro-3-deoxy-D-arabino-heptonate-7-phosphate D-erythrose-4-phosphate-lyase (pyruvate-phosphorylating), 2-dehydro-3-deoxy-phosphoheptanoate aldolase, 2-dehydro-3-deoxy-phosphoheptonate aldolase, 2-keto-3-deoxy-D-arabino-heptonic acid 7-phosphate synthetase, 3-deoxy-7-phosphoheptulonate synthase, 3-deoxy-D-arabino heptulosonate 7-phosphate synthase, 3-deoxy-D-arabino-2-heptulosonic acid 7-phosphate synthetase, 3-deoxy-D-arabino-heptolosonate-7-phosphate synthetase, 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase, 3-deoxy-D-arabino-heptulosonate 7-phosphate synthetase, 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase, 3-deoxy-D-arabino-heptulosonic acid 7-phosphate synthase, 3-deoxy-D-arabinoheptulosonate 7-phosphate synthase, 3-deoxy-D-arabinoheptulosonate-7-phosphate synthetase, 7-phospho-2-dehydro-3-deoxy-D-arabino-heptonate D-erythrose-4-phosphate-lyase (pyruvate-phosphorylating), 7-phospho-2-keto-3-deoxy-D-arabino-heptonate D-erythrose-4-phosphate lyase (pyruvate-phosphorylating), aldolase, phospho-2-keto-3-deoxyheptanoate, Aro1A, Aro4, Aro4p, AroA, AroA(Q), aroF, AroFFR, AroG, D-erythrose-4-phosphate-lyase, D-erythrose-4-phosphate-lyase (pyruvate-phosphorylating), DAH7-P synthase, DAH7-P synthase (phe), DAH7P, DAH7P synthase, DAH7PS, DAHP synthase, DAHP synthase-phe, DAHP synthase-trp, DAHP synthase-tyr, DAHP(Phe), DAHPS, DAHPS(Tyr), Dax1, deoxy-D-arabino-heptulosonate-7-phosphate synthetase, DHS, DHS1, DS-Co, DS-Mn, EC 4.1.2.15, KDPH synthase, KDPH synthetase, More, mtDAH7Ps, MtDS, NCgl0950 DAHP synthase, NCgl2098 DAHP synthase, Nme-DAH7PS, OsDAHPS, PF1690, phospho-2-dehydro-3-deoxyheptonate aldolase, phospho-2-keto-3-deoxyheptanoate aldolase, Phospho-2-keto-3-deoxyheptonate aldolase, phospho-2-keto-3-deoxyheptonic aldolase, phospho-2-oxo-3-deoxyheptonate aldolase, phzC, TmaDAH7PS

ECTree

     2 Transferases
         2.5 Transferring alkyl or aryl groups, other than methyl groups
             2.5.1 Transferring alkyl or aryl groups, other than methyl groups (only sub-subclass identified to date)
                2.5.1.54 3-deoxy-7-phosphoheptulonate synthase

Purification

Purification on EC 2.5.1.54 - 3-deoxy-7-phosphoheptulonate synthase

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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
2350fold purification of the tryptophan sensitive isozyme, partial purification of the tyrosine- and phenylalanine-sensitive isozyme
-
5-6fold to homogeneity
-
cells centrifuged and resuspended in 20 mM BTP (1,3-bis[tris(hydroxymethyl)methylamino]propane), pH 7.5, 150 mM NaCl, 0.5 mM TCEP (tris(2-carboxyethyl)phosphine hydrochloride), 0.2 mM phosphoenolpyruvate and 0.1 mM MnCl2, cell lysis with lysozyme and sonication, centrifugation, crude extract subjected to Ni-NTA agarose chromatography, elution with 250 mM imidazole in buffer, further purification by preparative FPLC HiLoad 26/60 Superdex 75 gel-filtration column
cytosolic isozyme, 79.8fold to homogeneity
-
enzyme III
-
His-Bind protein chromatography
-
Ni-NTA affinity chromatography followed by gel filtration, in 20 mM BTP (1,3-bis[tris(hydroxymethyl)methylamino]propane), pH 7.5, 150 mM NaCl, 0.5 mM TCEP [tris(2-carboxyethyl)phosphine hydrochloride], 0.2 mM phosphoenolpyruvate, and 0.1 mM MnCl2, ultrafiltration for concentration of enzyme
partial
partial, 19fold, recombinant from Escherichia coli
-
partial, isozyme DS-Co and isozyme DS-Mn
-
partially, 2 isozymes
-
partially, isozymes DS-Co and DS-Mn
-
Phe-sensitive isozyme, 11fold to homogeneity from overproducing strain
-
Phe-sensitive isozyme, recombinant wild-type from overexpressing strain and recombinant mutants
-
recombinant enzyme
-
recombinant from Escherichia coli, mature chloroplastic form, purified to homogeneity
-
recombinant from overexpressing strain
-
recombinant His-tagged wild-type and mutants from Escherichia coli
-
recombinant His10-tagged enzyme from Escherichia coli strain BL21 by affinity chromatography, followed by tag cleavage with factor Xa and again affinity chromatography and dialysis to remove the tag
recombinant mutant enzyme R126S from Escherichia coli strain BL21 StarTM (DE3)
recombinant protein
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21 (DE3) by anion exchange chromatography, ultrafiltration, and gel filtration
recombinantly overexpresed Tyr-sensitive isozyme, 7.5fold
-
strain JP1525
-
Trp-sensitive isozyme to homogeneity, 2000fold
-
Trp-sensitive isozyme to homogeneity, 2380fold, large scale
-
Trp-sensitive isozyme to homogeneity, 3750fold
-
Trp-sensitive isozyme, 130fold
-
Tyr-sensitive isozyme
-
Tyr-sensitive isozyme, wild-type and mutants from strain AB3257
-