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Literature summary extracted from

  • Schwarz, A.; Brecker, L.; Nidetzky, B.
    Acid-base catalysis in Leuconostoc mesenteroides sucrose phosphorylase probed by site-directed mutagenesis and detailed kinetic comparison of wild-type and Glu237-Gln mutant enzymes (2007), Biochem. J., 403, 441-449.
    View publication on PubMedView publication on EuropePMC

Protein Variants

EC Number Protein Variants Comment Organism
2.4.1.7 E237Q site-directed mutagenesis of the catalytic residue, the mutant shows 0.001% of wild-type enzyme activity, reactions with substrates requiring Broensted catalytic assistance for glucosylation or deglucosylation are selectively slowed at the respective step about 10fold in mutant E237Q compared to the wild-type enzyme. Azide, acetate and formate but not halides restore catalytic activity up to 300fold in E237Q under conditions in which the deglucosylation step is rate-determining Leuconostoc mesenteroides

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.1.7 additional information
-
additional information steady-state kinetics of wild-type and mutant E237Q enzymes Leuconostoc mesenteroides

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.4.1.7 55700
-
x * 55700, SDS-PAGE 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
-
-
-

Reaction

EC Number Reaction Comment Organism Reaction ID
2.4.1.7 sucrose + phosphate = D-fructose + alpha-D-glucose 1-phosphate reaction mechanism, acid-base catalysis in the two-step enzymatic mechanism of alpha-retaining glucosyl transfer by Leuconostoc mesenteroides sucrose phosphorylase, Glu237 is the catalytic acid-base of sucrose phosphorylase Leuconostoc mesenteroides

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.4.1.7 additional information
-
-
Leuconostoc mesenteroides
2.4.1.7 0.0017
-
recombinant mutant E237Q Leuconostoc mesenteroides

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.1.7 1-O-azido-alpha-D-glucose + phosphate
-
Leuconostoc mesenteroides ?
-
?
2.4.1.7 alpha-D-glucose 1-acetic acid ester + phosphate alpha-D-glucose 1-acetic acid ester is converted primarily into the alpha- and beta-anomers of 2-O-acetyl D-glucose Leuconostoc mesenteroides 2-O-acetyl D-glucose + ?
-
?
2.4.1.7 alpha-D-glucose 1-phosphate + arsenate
-
Leuconostoc mesenteroides ?
-
?
2.4.1.7 additional information regio- and stereoselective formation of alpha-glucose 1-acetic acid ester by mutant E237Q, NMR product determination, overview Leuconostoc mesenteroides ?
-
?
2.4.1.7 sucrose + phosphate
-
Leuconostoc mesenteroides D-fructose + alpha-D-glucose 1-phosphate
-
r
2.4.1.7 sucrose + phosphate enzyme deglucosylation to an anionic nucleophile takes place with Glu237 protonated or unprotonated. Enzymatically formed alpha-glucose 1-esters decompose spontaneously via acyl group migration and hydrolysis Leuconostoc mesenteroides D-fructose + alpha-D-glucose 1-phosphate
-
r

Subunits

EC Number Subunits Comment Organism
2.4.1.7 ? x * 55700, SDS-PAGE Leuconostoc mesenteroides

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 0.00063
-
alpha-D-glucose 1-phosphate pH 7.0, 30°C, synthesis, mutant E237Q Leuconostoc mesenteroides
2.4.1.7 0.0016
-
sucrose pH 7.0, 30°C, phosphorolysis, mutant E237Q 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 pH 7.0, 30°C, synthesis, wild-type enzyme Leuconostoc mesenteroides
2.4.1.7 165
-
sucrose pH 7.0, 30°C, phosphorolysis, wild-type enzyme Leuconostoc mesenteroides
2.4.1.7 195
-
phosphate pH 7.0, 30°C, phosphorolysis, wild-type enzyme Leuconostoc mesenteroides

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.1.7 additional information
-
deglucosylation of mutant E237Q is pH-independent Leuconostoc mesenteroides
2.4.1.7 5.6 7.2 optimal pH for glycosylation Leuconostoc mesenteroides