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Literature summary for 2.4.1.7 extracted from

  • Wildberger, P.; Luley-Goedl, C.; Nidetzky, B.
    Aromatic interactions at the catalytic subsite of sucrose phosphorylase: their roles in enzymatic glucosyl transfer probed with Phe52->Ala and Phe52->Asn mutants (2011), FEBS Lett., 585, 499-504.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of Strep-tagged wild-type and mutant enzymes in Escherichia coli Top10 cells Leuconostoc mesenteroides

Protein Variants

Protein Variants Comment Organism
F52A site-directed mutagenesis, large destabilization of the transition states for enzyme glucosylation and deglucosylation in the mutant compared to the wild-type enzyme, while the relative stability of the glucosyl enzyme intermediate was weakly affected by substitution of Phe52 Leuconostoc mesenteroides
F52N site-directed mutagenesis, large destabilization of the transition states for enzyme glucosylation and deglucosylation in the mutant compared to the wild-type enzyme, while the relative stability of the glucosyl enzyme intermediate was weakly affected by substitution of Phe52 Leuconostoc mesenteroides

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information free energy profiles for reactions of wild-type and mutated enzymes, and steady-state kinetic analysis, overview Leuconostoc mesenteroides
4.7
-
alpha-D-glucose 1-phosphate pH 7.0, 30°C, sucrose synthesis by glycosylation from alpha-D-glucose 1-phosphate, recombinant wild-type enzyme Leuconostoc mesenteroides
6
-
sucrose pH 7.0, 30°C, deglycosylation by phosphate, recombinant wild-type enzyme Leuconostoc mesenteroides
9.8
-
sucrose pH 7.0, 30°C, sucrose phosphorolysis, recombinant wild-type enzyme Leuconostoc mesenteroides
13
-
sucrose pH 7.0, 30°C, deglycosylation by D-fructose, recombinant wild-type enzyme Leuconostoc mesenteroides
22
-
alpha-D-glucopyranosyl fluoride pH 7.0, 30°C, phosphorolysis, recombinant wild-type enzyme Leuconostoc mesenteroides

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Leuconostoc mesenteroides
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant Strep-tagged wild-type and mutant enzymes from Escherichia coli Top10 cells by single step affinity chromatography Leuconostoc mesenteroides

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
alpha-D-glucopyranosyl fluoride + phosphate
-
Leuconostoc mesenteroides fluoride + alpha-D-glucose 1-phosphate
-
?
D-fructose + alpha-D-glucose 1-phosphate
-
Leuconostoc mesenteroides sucrose + phosphate
-
r
sucrose + phosphate
-
Leuconostoc mesenteroides D-fructose + alpha-D-glucose 1-phosphate
-
r

Synonyms

Synonyms Comment Organism
SPase
-
Leuconostoc mesenteroides

Temperature Optimum [°C]

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

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
39
-
alpha-D-glucose 1-phosphate pH 7.0, 30°C, sucrose synthesis by glycosylation from alpha-D-glucose 1-phosphate, recombinant wild-type enzyme Leuconostoc mesenteroides
47
-
sucrose pH 7.0, 30°C, deglycosylation by D-fructose, recombinant wild-type enzyme Leuconostoc mesenteroides
117
-
sucrose pH 7.0, 30°C, sucrose phosphorolysis, recombinant wild-type enzyme Leuconostoc mesenteroides
121
-
alpha-D-glucopyranosyl fluoride pH 7.0, 30°C, phosphorolysis, recombinant wild-type enzyme Leuconostoc mesenteroides
145
-
sucrose pH 7.0, 30°C, deglycosylation by phosphate, recombinant wild-type enzyme Leuconostoc mesenteroides

pH Optimum

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

General Information

General Information Comment Organism
evolution the enzyme is a transglucosidase belonging to glycosylhydrolase family GH 13 Leuconostoc mesenteroides

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.7
-
sucrose pH 7.0, 30°C, deglycosylation by D-fructose, recombinant wild-type enzyme Leuconostoc mesenteroides
5.5
-
alpha-D-glucopyranosyl fluoride pH 7.0, 30°C, phosphorolysis, recombinant wild-type enzyme Leuconostoc mesenteroides
8.3
-
alpha-D-glucose 1-phosphate pH 7.0, 30°C, sucrose synthesis by glycosylation from alpha-D-glucose 1-phosphate, recombinant wild-type enzyme Leuconostoc mesenteroides
12
-
sucrose pH 7.0, 30°C, sucrose phosphorolysis, recombinant wild-type enzyme Leuconostoc mesenteroides
24
-
sucrose pH 7.0, 30°C, deglycosylation by phosphate, recombinant wild-type enzyme Leuconostoc mesenteroides