Literature summary for 1.1.1.406 extracted from

Carius, Y.; Christian, H.; Faust, A.; Zander, U.; Klink, B.U.; Kornberger, P.; Kohring, G.W.; Giffhorn, F.; Scheidig, A.J.
Structural insight into substrate differentiation of the sugar-metabolizing enzyme galactitol dehydrogenase from Rhodobacter sphaeroides D (2010), J. Biol. Chem., 285, 20006-20014.

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Application

Application	Comment	Organism
synthesis	as an enzyme capable of the stereo- and regioselective modification of carbohydrates, GatDH exhibits a high potential for application in biotechnology as a biocatalyst, e.g. preparation of several (R)-1,2-diols by racemic resolution with GatDH as well as the synthesis of several S-configured aliphatic alcohols by reducing corresponding prochiral ketones	Cereibacter sphaeroides

Cloned(Commentary)

Cloned (Comment)	Organism
expression of His6-tagged GatDH in Escherichia coli strain BL21(DE3)	Cereibacter sphaeroides

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant enzyme, free or with bound 1,2-pentanediol or meso-erythritol, hanging drop vapour diffusion method, 0.001 ml of 0.45 mM protein with 1 mM NAD+ or NADH, in 20 mM Bis-Tris, pH 6.5, is mixed with 0.001 ml of reservoir solution containing 100 mM MES, pH 5.5-5.9, 200 mM MgCl2, and 10-20% w/v methoxy poly(ethylene glycol), equilibration against 1 ml reservoir solution, 1 week, X-ray diffraction structure determnination and analysis at 1.25-195 A resolution	Cereibacter sphaeroides

Crystallization (Comment)

Organism

purified recombinant enzyme, free or with bound 1,2-pentanediol or meso-erythritol, hanging drop vapour diffusion method, 0.001 ml of 0.45 mM protein with 1 mM NAD+ or NADH, in 20 mM Bis-Tris, pH 6.5, is mixed with 0.001 ml of reservoir solution containing 100 mM MES, pH 5.5-5.9, 200 mM MgCl2, and 10-20% w/v methoxy poly(ethylene glycol), equilibration against 1 ml reservoir solution, 1 week, X-ray diffraction structure determnination and analysis at 1.25-195 A resolution

Cereibacter sphaeroides

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
22	-	galactitol	pH not specified in the publication, temperature not specified in the publication	Cereibacter sphaeroides

Metals/Ions

Metals/Ions	Comment	Organism
Co2+	can replace Mg2+	Cereibacter sphaeroides
Mg2+	GatDH constitutes a homotetramer with two magnesium-ion binding sites each formed by two opposing C termini, binding site structure, overview	Cereibacter sphaeroides
additional information	activity of GatDH is strictly dependent on the presence of divalent metal ions	Cereibacter sphaeroides

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
galactitol + NAD+	Cereibacter sphaeroides	-	L-tagatose + NADH + H+	-	r
additional information	Cereibacter sphaeroides	GatDH catalyzes the dehydrogenation of a variety of polyvalent aliphatic alcohols and polyols to the corresponding ketones and ketoses, respectively, and in the reverse reaction it reduces prochiral ketones with high stereoselectivity yielding the corresponding S-configured secondary alcohols	?	-	?
additional information	Cereibacter sphaeroides D	GatDH catalyzes the dehydrogenation of a variety of polyvalent aliphatic alcohols and polyols to the corresponding ketones and ketoses, respectively, and in the reverse reaction it reduces prochiral ketones with high stereoselectivity yielding the corresponding S-configured secondary alcohols	?	-	?
xylitol + NAD+	Cereibacter sphaeroides	-	L-xylulose + NADH + H+	-	r
xylitol + NAD+	Cereibacter sphaeroides D	-	L-xylulose + NADH + H+	-	r

Organism

Organism	UniProt	Comment	Textmining
Cereibacter sphaeroides	C0KTJ6	-	-
Cereibacter sphaeroides D	C0KTJ6	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His6-tagged GatDH from Escherichia coli strain BL21(DE3), by nickel affinity chromatography the tag is cleaved off, followed by dialysis and gel filtration	Cereibacter sphaeroides

Reaction

Reaction	Comment	Organism	Reaction ID
galactitol + NAD+ = D-tagatose + NADH + H+	reaction mechanism, overview	Cereibacter sphaeroides	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
1,2-(S)-pentanediol + NAD+	-	Cereibacter sphaeroides	? + NADH + H+	-	r
galactitol + NAD+	-	Cereibacter sphaeroides	L-tagatose + NADH + H+	-	r
meso-erythritol + NAD+	very low activity	Cereibacter sphaeroides	? + NADH + H+	-	r
meso-erythritol + NAD+	very low activity	Cereibacter sphaeroides D	? + NADH + H+	-	r
additional information	GatDH catalyzes the dehydrogenation of a variety of polyvalent aliphatic alcohols and polyols to the corresponding ketones and ketoses, respectively, and in the reverse reaction it reduces prochiral ketones with high stereoselectivity yielding the corresponding S-configured secondary alcohols	Cereibacter sphaeroides	?	-	?
additional information	site mapping, the catalytic tetrad is formed by Asn116, Ser144, Tyr159, and Lys163. The substrate binding pocket can be divided into two parts of different size and polarity. In the smaller part, the side chains of amino acids Ser144, Ser146, and Asn151 are important determinants for the binding specificity and the orientation of (pro-) chiral compounds. The larger part of the pocket is elongated and flanked by polar and non-polar residues, enabling a rather broad substrate spectrum. NAD(H) binding structure, overview	Cereibacter sphaeroides	?	-	?
additional information	GatDH catalyzes the dehydrogenation of a variety of polyvalent aliphatic alcohols and polyols to the corresponding ketones and ketoses, respectively, and in the reverse reaction it reduces prochiral ketones with high stereoselectivity yielding the corresponding S-configured secondary alcohols	Cereibacter sphaeroides D	?	-	?
additional information	site mapping, the catalytic tetrad is formed by Asn116, Ser144, Tyr159, and Lys163. The substrate binding pocket can be divided into two parts of different size and polarity. In the smaller part, the side chains of amino acids Ser144, Ser146, and Asn151 are important determinants for the binding specificity and the orientation of (pro-) chiral compounds. The larger part of the pocket is elongated and flanked by polar and non-polar residues, enabling a rather broad substrate spectrum. NAD(H) binding structure, overview	Cereibacter sphaeroides D	?	-	?
xylitol + NAD+	-	Cereibacter sphaeroides	L-xylulose + NADH + H+	-	r
xylitol + NAD+	410% of the activity with galactitol	Cereibacter sphaeroides	L-xylulose + NADH + H+	-	r
xylitol + NAD+	-	Cereibacter sphaeroides D	L-xylulose + NADH + H+	-	r
xylitol + NAD+	410% of the activity with galactitol	Cereibacter sphaeroides D	L-xylulose + NADH + H+	-	r

Subunits

Subunits	Comment	Organism
homotetramer	-	Cereibacter sphaeroides

Synonyms

Synonyms	Comment	Organism
GatDH	-	Cereibacter sphaeroides
More	GatDH belongs to the protein superfamily of short-chain dehydrogenases	Cereibacter sphaeroides

Cofactor

Cofactor	Comment	Organism	Structure
NAD+	-	Cereibacter sphaeroides
NADH	-	Cereibacter sphaeroides