Literature summary for 2.4.1.13 extracted from

Wu, R.; Asencion Diez, M.D.; Figueroa, C.M.; Machtey, M.; Iglesias, A.A.; Ballicora, M.A.; Liu, D.
The crystal structure of Nitrosomonas europaea sucrose synthase reveals critical conformational changes and insights into sucrose metabolism in prokaryotes (2015), J. Bacteriol., 197, 2734-2746.

Cloned(Commentary)

Cloned (Comment)	Organism
gene ss2, phylogenetic analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) using plasmid pNESS2	Nitrosomonas europaea

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant His-tagged enzyme, hanging drop vapour diffusion method, mixing of 0.001 ml of 15mg/ml protein solution with 0.001 ml of reservoir solution containing 5% Tacsimate, pH 5.0, 5% w/v PEG 3350, and 0.1 M sodium citrate, pH 5.6, 20°C, 7 days, X-ray diffraction structure determination and analysis at 3.05 A resolution, molecular replacement with Arabidopsis thaliana sucrose synthase structural model, PDB ID 3S29, as a template	Nitrosomonas europaea

Crystallization (Comment)

Organism

purified recombinant His-tagged enzyme, hanging drop vapour diffusion method, mixing of 0.001 ml of 15mg/ml protein solution with 0.001 ml of reservoir solution containing 5% Tacsimate, pH 5.0, 5% w/v PEG 3350, and 0.1 M sodium citrate, pH 5.6, 20°C, 7 days, X-ray diffraction structure determination and analysis at 3.05 A resolution, molecular replacement with Arabidopsis thaliana sucrose synthase structural model, PDB ID 3S29, as a template

Nitrosomonas europaea

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.044	-	ADP-glucose	pH 7.0, 60°C	Nitrosomonas europaea
0.89	-	UDP-glucose	pH 7.0, 60°C	Nitrosomonas europaea
5.6	-	D-fructose	pH 7.0, 60°C, with ADP-glucose	Nitrosomonas europaea
120	-	D-fructose	pH 7.0, 60°C, with UDP-glucose	Nitrosomonas europaea

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
95000	-	4 * 95000, recombinant His-tagged enzyme, SDS-PAGE, each monomer contains four domains, i.e. SSN-1, SSN-2, GT-B(A), and GT-B(D), structure overview and comparisons	Nitrosomonas europaea
360000	-	recombinant His-tagged enzyme, gel filtration	Nitrosomonas europaea

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
ADP-glucose + D-fructose	Nitrosomonas europaea	-	ADP + sucrose	-	r
NDP-glucose + D-fructose	Nitrosomonas europaea	-	NDP + sucrose	-	r
UDP-glucose + D-fructose	Nitrosomonas europaea	-	UDP + sucrose	-	r

Organism

Organism	UniProt	Comment	Textmining
Nitrosomonas europaea	Q820M5	gene ss2	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration to homogeneity	Nitrosomonas europaea

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
ADP-glucose + D-fructose	-	Nitrosomonas europaea	ADP + sucrose	-	r
NDP-glucose + D-fructose	-	Nitrosomonas europaea	NDP + sucrose	-	r
NDP-glucose + D-fructose	the enzyme prefers transferring glucose from ADP-glucose rather than UDP-glucose	Nitrosomonas europaea	NDP + sucrose	-	r
UDP-glucose + D-fructose	-	Nitrosomonas europaea	UDP + sucrose	-	r

Subunits

Subunits	Comment	Organism
homotetramer	4 * 95000, recombinant His-tagged enzyme, SDS-PAGE, each monomer contains four domains, i.e. SSN-1, SSN-2, GT-B(A), and GT-B(D), structure overview and comparisons	Nitrosomonas europaea

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Nitrosomonas europaea

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8	-	assay at	Nitrosomonas europaea

General Information

General Information	Comment	Organism
evolution	Nitrosomonas europaea sucrose synthase shares the same fold as the GT-B family of the retaining glycosyltransferases	Nitrosomonas europaea
additional information	a triad of conserved homologous catalytic residues, Arg567, Lys572, and Glu663, in the family is functionally critical in the Nitrosomonas europaea sucrose synthase, structure homology modeling, overview	Nitrosomonas europaea

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
0.036	-	D-fructose	pH 7.0, 60°C, with UDP-glucose	Nitrosomonas europaea
1.33	-	D-fructose	pH 7.0, 60°C, with ADP-glucose	Nitrosomonas europaea
7.5	-	ADP-glucose	pH 7.0, 60°C	Nitrosomonas europaea
130.3	-	UDP-glucose	pH 7.0, 60°C	Nitrosomonas europaea