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Literature summary 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.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

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

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.4.1.13 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.1.13 0.044
-
 ADP-glucose pH 7.0, 60°C Nitrosomonas europaea
2.4.1.13 0.89
-
 UDP-glucose pH 7.0, 60°C Nitrosomonas europaea
2.4.1.13 5.6
-
 D-fructose pH 7.0, 60°C, with ADP-glucose Nitrosomonas europaea
2.4.1.13 120
-
 D-fructose pH 7.0, 60°C, with UDP-glucose Nitrosomonas europaea

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.4.1.13 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
2.4.1.13 360000
-
 recombinant His-tagged enzyme, gel filtration Nitrosomonas europaea

Natural Substrates/ Products (Substrates)

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

Organism

EC Number Organism UniProt Comment Textmining
2.4.1.13 Nitrosomonas europaea Q820M5 gene ss2
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.4.1.13 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)

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

Subunits

EC Number Subunits Comment Organism
2.4.1.13 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]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.1.13 37
-
 assay at Nitrosomonas europaea

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.1.13 8
-
 assay at Nitrosomonas europaea

General Information

EC Number General Information Comment Organism
2.4.1.13 evolution Nitrosomonas europaea sucrose synthase shares the same fold as the GT-B family of the retaining glycosyltransferases Nitrosomonas europaea
2.4.1.13 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]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.4.1.13 0.036
-
 D-fructose pH 7.0, 60°C, with UDP-glucose Nitrosomonas europaea
2.4.1.13 1.33
-
 D-fructose pH 7.0, 60°C, with ADP-glucose Nitrosomonas europaea
2.4.1.13 7.5
-
 ADP-glucose pH 7.0, 60°C Nitrosomonas europaea
2.4.1.13 130.3
-
 UDP-glucose pH 7.0, 60°C Nitrosomonas europaea