1.1.1.25: shikimate dehydrogenase

This is an abbreviated version, for detailed information about shikimate dehydrogenase, go to the full flat file.

Reaction

shikimate
+
NADP+
=
3-dehydroshikimate
+
NADPH
+
H+

Synonyms

3-dehydroquinate dehydratase/shikimate dehydrogenase, 5-dehydroshikimate reductase, ael1, Af2327, AroE, cgR_0495, cgR_1677, dehydroquinate dehydratase-shikimate dehydrogenase, dehydroshikimic reductase, DHD/SHD, DHQ-SDH, HI0607, More, MtbSD, NADP-dependent shikimate dehydrogenase, qsuD, rifI, SD, SDH, sdhL, shikimate 5-dehydrogenase, shikimate dehydrogenase, shikimate oxidoreductase, shikimate:NADP oxidoreductase, shikimate:NADP+ 5-oxidoreductase, SKDH, tm0346

ECTree

     1 Oxidoreductases
         1.1 Acting on the CH-OH group of donors
             1.1.1 With NAD+ or NADP+ as acceptor
                1.1.1.25 shikimate dehydrogenase

Purification

Purification on EC 1.1.1.25 - shikimate dehydrogenase

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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
; recombinant enzyme from aroE auxotrophic mutant strain AB2834 by ammonium sulfate fractionation, anion exchange chromatography, ultrafiltration, and gel filtration
-
by nickel-nitrilotriacetic acid affinity chromatography; by nickel-nitrilotriacetic acid affinity chromatography
Cultures with expressed SDH enzymes are incubated and then induced with 0.4 mM isopropyl-beta-D-thiogalactopyranoside. Harvested cells are disrupted and the insoluble cellular material is removed by centrifugation. The recombinants are purified from other contaminating proteins using nickel-nitrilotriacetic acid affinity chromatography. Protein samples for kinetic studies are dialyzed and stored at 4 °C in 10 mM Tris–HCl, 500 mM NaCl, and 5% glycerol.; Cultures with expressed SDH enzymes are incubated and then induced with 0.4 mM isopropyl-beta-D-thiogalactopyranoside. Harvested cells are disrupted and the insoluble cellular material is removed by centrifugation. The recombinants are purified from other contaminating proteins using nickel-nitrilotriacetic acid affinity chromatography. Protein samples for kinetic studies are dialyzed and stored at 4 °C in 10 mM Tris–HCl, 500 mM NaCl, and 5% glycerol.; Cultures with expressed SDH enzymes are incubated and then induced with 0.4 mM isopropyl-beta-D-thiogalactopyranoside. Harvested cells are disrupted and the insoluble cellular material is removed by centrifugation. The recombinants are purified from other contaminating proteins using nickel-nitrilotriacetic acid affinity chromatography. Protein samples for kinetic studies are dialyzed and stored at 4°C in 10 mM Tris-HCl, 500 mM NaCl, and 5% glycerol.; Cultures with expressed SDH enzymes are incubated and then induced with 0.4 mM isopropyl-beta-D-thiogalactopyranoside. Harvested cells are disrupted and the insoluble cellular material is removed by centrifugation. The recombinants are purified from other contaminating proteins using nickel-nitrilotriacetic acid affinity chromatography. Protein samples for kinetic studies are dialyzed and stored at 4°C in 10 mM Tris–HCl, 500 mM NaCl, and 5% glycerol.; Cultures with expressed SDH enzymes are incubated and then induced with 0.4 mM isopropyl-beta-D-thiogalactopyranoside. Harvested cells are disrupted and the insoluble cellular material is removed by centrifugation. The recombinants are purified from other contaminating proteins using nickel-nitrilotriacetic acid affinity chromatography. Protein samples for kinetic studies are dialyzed and stored at 4°C in 10 mM Tris–HCl, 500 mM NaCl, and 5% glycerol.
enzyme complex of EC 4.2.1.10 and EC 1.1.1.25
-
homogeneous recombinant mutant K69A and wild-type proteins are obtained by a three-step purification protocol, by gel filtration, 11.8fold
-
multienzyme complex
-
one-step purification by nickel-affinity chromatography
recombinant enzyme expressed in Escherichia coli, formation of insoluble aggregates, solubilization by repeated freezing and thawing
-
recombinant GST-tagged SeMet-labeled enzyme from Escherichia coli strain BL21 by glutathione affinity and ion exchange chromatography, proteolytic removal of the GST-tag
recombinant His-tagged ARoE from Escherichia coli strain BL21(DE3) by nickel affinity chromatograohy
-
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3)
-
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
recombinant protein; recombinant protein
recombinant soluble enzyme 8.5fold from Escherichia coli strain BL21(DE3) by ion exchange and hydrophobic interaction chromatography, gel filtration, and again ion exchange chromatography, to homogeneity
separation of isoenzymes by regaining the protein from PA-gel slices
-