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Literature summary extracted from

  • Mueller, M.; Nidetzky, B.
    The role of Asp-295 in the catalytic mechanism of Leuconostoc mesenteroides sucrose phosphorylase probed with site-directed mutagenesis (2007), FEBS Lett., 581, 1403-1408.
    View publication on PubMed

Protein Variants

EC Number Protein Variants Comment Organism
2.4.1.7 D295E mutation decreases the catalytic center activity of sucrose phosphorylase to about 0.01% of the wild-type level. The 100000fold preference of the wild-type for glucosyl transfer compared with mannosyl transfer from phosphate to fructose is lost Leuconostoc mesenteroides
2.4.1.7 D295E site-directed mutagenesis of the catalytic residue, the mutant shows about 0.01% of the wild-type enzyme activity, the preference of the wild-type enzyme for glucosyl transfer compared with mannosyl transfer from phosphate to fructose is lost in the mutant enzyme Leuconostoc mesenteroides
2.4.1.7 D295N mutation decreases the catalytic center activity of sucrose phosphorylase to about 0.01% of the wild-type level. Glucosylation and deglucosylation steps are affected uniformly, and independently of leaving group ability and nucleophilic reactivity of the substrate, respectively. The 100000fold preference of the wild-type for glucosyl transfer compared with mannosyl transfer from phosphate to fructose is lost Leuconostoc mesenteroides
2.4.1.7 D295N site-directed mutagenesis of the catalytic residue, the mutant shows about 0.01% of the wild-type enzyme activity, the preference of the wild-type enzyme for glucosyl transfer compared with mannosyl transfer from phosphate to fructose is lost in the mutant enzyme Leuconostoc mesenteroides

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.4.1.7 sucrose + phosphate Leuconostoc mesenteroides
-
 D-fructose + alpha-D-glucose 1-phosphate
-
 r

Organism

EC Number Organism UniProt Comment Textmining
2.4.1.7 Leuconostoc mesenteroides
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.4.1.7 mutant enzymes D295N and D295E Leuconostoc mesenteroides

Reaction

EC Number Reaction Comment Organism Reaction ID
2.4.1.7 sucrose + phosphate = D-fructose + alpha-D-glucose 1-phosphate reaction mechanism, the side chain of Asp295, through a strong hydrogen bond with the equatorial sugar 2-hydroxyl, stabilizes the transition states flanking the bglucosyl enzyme intermediate Leuconostoc mesenteroides
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.1.7 alpha-D-glucose 1-phosphate + arsenate
-
 Leuconostoc mesenteroides ?
-
 ?
2.4.1.7 additional information glucosyl donor and acceptor specificities, in absence of acceptor, the enzyme performs hydrolysis of alpha-D-glucose 1-phosphate Leuconostoc mesenteroides ?
-
 ?
2.4.1.7 sucrose + phosphate
-
 Leuconostoc mesenteroides D-fructose + alpha-D-glucose 1-phosphate
-
 r

Synonyms

EC Number Synonyms Comment Organism
2.4.1.7 SPase
-
 Leuconostoc mesenteroides

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.1.7 30
-
 assay at Leuconostoc mesenteroides

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.4.1.7 additional information
-
 additional information kinetics of recombinant wild-type and mutant enzymes, overview Leuconostoc mesenteroides
2.4.1.7 0.0014
-
 D-fructose pH 7.0, 30°C, synthesis, mutant D295E Leuconostoc mesenteroides
2.4.1.7 0.0065
-
 arsenate pH 7.0, 30°C, arsenolysis, mutant D295E Leuconostoc mesenteroides
2.4.1.7 0.0124
-
 alpha-D-glucose 1-phosphate pH 7.0, 30°C, synthesis, mutant D295E Leuconostoc mesenteroides
2.4.1.7 0.0136
-
 phosphate pH 7.0, 30°C, phosphorolysis, mutant D295E Leuconostoc mesenteroides
2.4.1.7 0.0159
-
 alpha-D-glucose 1-phosphate pH 7.0, 30°C, synthesis, mutant D295N Leuconostoc mesenteroides
2.4.1.7 0.02
-
 sucrose pH 7.0, 30°C, phosphorolysis, mutant D295N Leuconostoc mesenteroides
2.4.1.7 0.0206
-
 D-fructose pH 7.0, 30°C, synthesis, mutant D295N Leuconostoc mesenteroides
2.4.1.7 0.024
-
 phosphate pH 7.0, 30°C, phosphorolysis, mutant D295N Leuconostoc mesenteroides
2.4.1.7 0.0252
-
 arsenate pH 7.0, 30°C, arsenolysis, mutant D295N Leuconostoc mesenteroides
2.4.1.7 1.4
-
 D-fructose 30°C, pH 7.0, mutant enzyme D295E Leuconostoc mesenteroides
2.4.1.7 1.59
-
 alpha-D-glucose 1-phosphate 30°C, pH 7.0, mutant enzyme D295N Leuconostoc mesenteroides
2.4.1.7 1.99
-
 sucrose 30°C, pH 7.0, mutant enzyme D295N Leuconostoc mesenteroides
2.4.1.7 2.06
-
 D-fructose 30°C, pH 7.0, mutant enzyme D295N Leuconostoc mesenteroides
2.4.1.7 2.38
-
 phosphate 30°C, pH 7.0, mutant enzyme D295N Leuconostoc mesenteroides
2.4.1.7 12.4
-
 alpha-D-glucose 1-phosphate 30°C, pH 7.0, mutant enzyme D295E Leuconostoc mesenteroides
2.4.1.7 13.6
-
 phosphate 30°C, pH 7.0, mutant enzyme D295E Leuconostoc mesenteroides
2.4.1.7 72
-
 D-fructose 30°C, pH 7.0, wild-type enzyme Leuconostoc mesenteroides
2.4.1.7 72
-
 D-fructose pH 7.0, 30°C, synthesis, wild-type enzyme Leuconostoc mesenteroides
2.4.1.7 91
-
 alpha-D-glucose 1-phosphate 30°C, pH 7.0, wild-type enzyme Leuconostoc mesenteroides
2.4.1.7 91
-
 alpha-D-glucose 1-phosphate pH 7.0, 30°C, synthesis, wild-type enzyme Leuconostoc mesenteroides
2.4.1.7 165
-
 sucrose 30°C, pH 7.0, wild-type enzyme Leuconostoc mesenteroides
2.4.1.7 165
-
 sucrose pH 7.0, 30°C, phosphorolysis, wild-type enzyme Leuconostoc mesenteroides
2.4.1.7 170
-
 arsenate pH 7.0, 30°C, arsenolysis, wild-type enzyme Leuconostoc mesenteroides
2.4.1.7 195
-
 phosphate 30°C, pH 7.0, wild-type enzyme Leuconostoc mesenteroides
2.4.1.7 195
-
 phosphate pH 7.0, 30°C, phosphorolysis, wild-type enzyme Leuconostoc mesenteroides
2.4.1.7 261
-
 alpha-D-glucose 1-phosphate pH 7.0, 30°C, arsenolysis, wild-type enzyme Leuconostoc mesenteroides

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.1.7 7
-
 assay at Leuconostoc mesenteroides

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.4.1.7 additional information
-
 pH-dependence of wild-type and mutant enzymes, overview Leuconostoc mesenteroides