Literature summary for 4.2.3.4 extracted from

Ngo, P.H.; Natarajan, S.; Kim, H.; Hung, H.K.; Kim, J.; Lee, B.; Ahn, Y.; Kang, L.
Cloning, expression, crystallization and preliminary x-ray crystallographic analysis of 3-dehydroquinate synthase, Xoo1243, from Xanthomonas oryzae pv. oryzae (2008), Acta Crystallogr. Sect. F, 64, 1128-1131.

Application

Application	Comment	Organism
drug development	potential antibiotic target	Xanthomonas oryzae

Cloned(Commentary)

Cloned (Comment)	Organism
The aroB gene cloned from Xoo and the corresponding DHQS protein is subsequently overexpressed in Escherichia coli	Xanthomonas oryzae

Crystallization (Commentary)

Crystallization (Comment)	Organism
by using the hanging-drop vapour-diffusion method with different screening kits, yields crystals belonging to the tetragonal space group, crystals of DHQS obtained in sitting-drop setups using a hydra automated high-throughput crystal screening machine with different screening kits: (a) 0.1 M sodium malonate pH 4.0, 12% PEG 3350, (b) 0.2 M sodium malonate pH 6.0, 20% PEG 3350, (c) 0.1 M Tris pH 8.5, 2.0 M ammonium sulfate, (d) 0.15 M caesium chloride, 15% PEG 3350, (e) 1.0 M sodium citrate, 0.1 M CHES pH 9.5, (f) 0.2 M ammonium sulfate, 0.1 M bis-Tris pH 5.5, 25% PEG 3350, (g) 0.2 M ammonium sulfate, 0.1 M MES pH 6.5, 30% PEG MME 5000, (h) 0.02 M zinc chloride, 20% PEG 3350. Best crystals are obtained after two weeks from a reservoir containing 0.2 M ammonium sulfate, 0.1 M MES monohydrate pH 6.0 and 31%PEG MME 5000	Xanthomonas oryzae

Crystallization (Comment)

Organism

by using the hanging-drop vapour-diffusion method with different screening kits, yields crystals belonging to the tetragonal space group, crystals of DHQS obtained in sitting-drop setups using a hydra automated high-throughput crystal screening machine with different screening kits: (a) 0.1 M sodium malonate pH 4.0, 12% PEG 3350, (b) 0.2 M sodium malonate pH 6.0, 20% PEG 3350, (c) 0.1 M Tris pH 8.5, 2.0 M ammonium sulfate, (d) 0.15 M caesium chloride, 15% PEG 3350, (e) 1.0 M sodium citrate, 0.1 M CHES pH 9.5, (f) 0.2 M ammonium sulfate, 0.1 M bis-Tris pH 5.5, 25% PEG 3350, (g) 0.2 M ammonium sulfate, 0.1 M MES pH 6.5, 30% PEG MME 5000, (h) 0.02 M zinc chloride, 20% PEG 3350. Best crystals are obtained after two weeks from a reservoir containing 0.2 M ammonium sulfate, 0.1 M MES monohydrate pH 6.0 and 31%PEG MME 5000

Xanthomonas oryzae

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
3-deoxy-D-arabinoheptulosonate 7-phosphate	Xanthomonas oryzae	shikimate pathway is present in all organisms except mammals	3-dehydroquinate + phosphate	-	?
additional information	Xanthomonas oryzae	enzyme plays an essential role in the synthesis of aromatic compounds in the shikimate pathway	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Xanthomonas oryzae	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
by Ni2+-NTA affinity chromatography, enzyme is subjected to a TEV protease cleavage reaction to cleave the six His-tag residues at the N-terminus, further purification by ion-exchange chromatography	Xanthomonas oryzae

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
additional information	-	Inactivation of enzymes in the shikimate pathway results in attenuation of virulence and loss of viability in a large number of bacterial species	Xanthomonas oryzae

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
3-deoxy-D-arabinoheptulosonate 7-phosphate	shikimate pathway is present in all organisms except mammals	Xanthomonas oryzae	3-dehydroquinate + phosphate	-	?
additional information	enzyme plays an essential role in the synthesis of aromatic compounds in the shikimate pathway	Xanthomonas oryzae	?	-	?

Synonyms

Synonyms	Comment	Organism
3-dehydroquinate synthase	-	Xanthomonas oryzae
DHQS	-	Xanthomonas oryzae
Xoo1243	-	Xanthomonas oryzae

Cofactor

Cofactor	Comment	Organism	Structure
NAD+	-	Xanthomonas oryzae