Literature summary for 5.3.99.10 extracted from

Hazra, A.B.; Han, Y.; Chatterjee, A.; Zhang, Y.; Lai, R.Y.; Ealick, S.E.; Begley, T.P.
A missing enzyme in thiamin thiazole biosynthesis: identification of TenI as a thiazole tautomerase (2011), J. Am. Chem. Soc., 133, 9311-9319.

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Cloned(Commentary)

Cloned (Comment)	Organism
gene tenI, overexpression as His-tagged enzyme in Escherichia coli strain BL21(DE3)	Bacillus subtilis

Crystallization (Commentary)

Crystallization (Comment)	Organism
enzyme TenI complexed with its reaction product cThz-P bound in the active site, hanging drop vapor diffusion method, liand-free TenI is grown from 1.65 to 1.75 M ammonium sulfate, 100 mM bicine, pH 8.7, 9.6, 2% PEG 400 w/v, and 8 mM L-cysteine, crystals are first dialyzed into 2.38 M sodium malonate, pH 7.0, to remove the sulfate ions by gradually increasing the sodium malonate concentration and decreasing the ammonium sulfate concentration, and soaked overnight in 2.38 M sodium malonate, 21.5 mM bicine, pH 9.0, 0.01% PEG400 w/v, 0.5 mM L-cysteine, 4% glycerol, and 11.5 mM cThz-P, X-ray diffraction structure determination and analysis	Bacillus subtilis

Crystallization (Comment)

Organism

enzyme TenI complexed with its reaction product cThz-P bound in the active site, hanging drop vapor diffusion method, liand-free TenI is grown from 1.65 to 1.75 M ammonium sulfate, 100 mM bicine, pH 8.7, 9.6, 2% PEG 400 w/v, and 8 mM L-cysteine, crystals are first dialyzed into 2.38 M sodium malonate, pH 7.0, to remove the sulfate ions by gradually increasing the sodium malonate concentration and decreasing the ammonium sulfate concentration, and soaked overnight in 2.38 M sodium malonate, 21.5 mM bicine, pH 9.0, 0.01% PEG400 w/v, 0.5 mM L-cysteine, 4% glycerol, and 11.5 mM cThz-P, X-ray diffraction structure determination and analysis

Bacillus subtilis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
2-[(2R,5Z)-2-carboxy-4-methylthiazol-5(2H)-ylidene]ethyl phosphate	Bacillus subtilis	-	2-(2-carboxy-4-methylthiazol-5-yl)ethyl phosphate	-	ir

Organism

Organism	UniProt	Comment	Textmining
Bacillus subtilis	-	gene tenI	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged TenI from Escherichia coli strain BL21(DE3) by nickel affinity chromatography	Bacillus subtilis

Reaction

Reaction	Comment	Organism	Reaction ID
2-[(2R,5Z)-2-carboxy-4-methylthiazol-5(2H)-ylidene]ethyl phosphate = 2-(2-carboxy-4-methylthiazol-5-yl)ethyl phosphate	reaction mechanism, overview	Bacillus subtilis	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
2-[(2R,5Z)-2-carboxy-4-methylthiazol-5(2H)-ylidene]ethyl phosphate	-	Bacillus subtilis	2-(2-carboxy-4-methylthiazol-5-yl)ethyl phosphate	-	ir
2-[(2R,5Z)-2-carboxy-4-methylthiazol-5(2H)-ylidene]ethyl phosphate	reversibility analysis by NMR	Bacillus subtilis	2-(2-carboxy-4-methylthiazol-5-yl)ethyl phosphate	-	ir

Synonyms

Synonyms	Comment	Organism
tenI	-	Bacillus subtilis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.6	-	assay at	Bacillus subtilis

General Information

General Information	Comment	Organism
metabolism	TenI is involved in the thiamin thiazole biosynthesis. In Bacillus subtilis, the thiazole tautomer (R,Z)-2-(2-carboxy-4-methylthiazol-5(2H)-ylidene)ethyl phosphate is formed by an oxidative condensation of glycine, 1-deoxy-D-xylulose-5-phosphate, and cysteine	Bacillus subtilis
physiological function	TenI shows no thiamin phosphate synthase activity, but functionsas a thiazole tautomerase, substrates phosphate and histidine 122 act as the acid/base residues involved in catalysis, reaction mechanism, overview. TenI does not affect glycine oxidase, thiamin phosphate synthase, or ThiS-COSH formation	Bacillus subtilis