Cloned (Comment) | Organism |
---|---|
gene dsrS, sequence comparisons and phylogenetic analysis, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21 AI | Leuconostoc mesenteroides |
Protein Variants | Comment | Organism |
---|---|---|
D306X | random mutagenesis | Leuconostoc mesenteroides |
D460A/H463S/T464L | site-directed mutagenesis | Leuconostoc mesenteroides |
D460M/H463Y/T464M/S512C | site-directed mutagenesis | Leuconostoc mesenteroides |
D460X | random mutagenesis | Leuconostoc mesenteroides |
F353T | random mutagenesis | Leuconostoc mesenteroides |
F353W | random mutagenesis | Leuconostoc mesenteroides |
F353X | random mutagenesis | Leuconostoc mesenteroides |
H463R/T464D/S512T | site-directed mutagenesis | Leuconostoc mesenteroides |
H463R/T464V/S512T | site-directed mutagenesis | Leuconostoc mesenteroides |
H463X | random mutagenesis | Leuconostoc mesenteroides |
additional information | screening of diverse mutants of the eight conserved residues that are determined to be important for enzyme activity, overview. Construction of enzyme mutant DSR-S vardel DELTA4N | Leuconostoc mesenteroides |
N404X | random mutagenesis | Leuconostoc mesenteroides |
S512C | random mutagenesis | Leuconostoc mesenteroides |
S512X | random mutagenesis | Leuconostoc mesenteroides |
T464X | random mutagenesis | Leuconostoc mesenteroides |
W440X | random mutagenesis | Leuconostoc mesenteroides |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.5 | - |
sucrose | pH 5.2, 25°C, recombinant mutant F353W | Leuconostoc mesenteroides | |
3.9 | - |
sucrose | pH 5.2, 25°C, recombinant mutant S512C | Leuconostoc mesenteroides | |
5.3 | - |
sucrose | pH 5.2, 25°C, recombinant mutant D460A/H463S/T464L | Leuconostoc mesenteroides | |
6.9 | - |
sucrose | pH 5.2, 25°C, recombinant mutant H463R/T464V/S512T | Leuconostoc mesenteroides | |
15.5 | - |
sucrose | pH 5.2, 25°C, recombinant mutant D460M/H463Y/T464M/S512C | Leuconostoc mesenteroides | |
15.5 | - |
sucrose | pH 5.2, 25°C, recombinant mutant H463R/T464D/S512T | Leuconostoc mesenteroides | |
115.8 | - |
sucrose | pH 5.2, 25°C, recombinant mutant F353T | Leuconostoc mesenteroides |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | required | Leuconostoc mesenteroides |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
sucrose + [(1->6)-alpha-D-glucosyl]n | Leuconostoc mesenteroides | - |
D-fructose + [(1->6)-alpha-D-glucosyl]n+1 | - |
? | |
sucrose + [(1->6)-alpha-D-glucosyl]n | Leuconostoc mesenteroides NRRL B-512F | - |
D-fructose + [(1->6)-alpha-D-glucosyl]n+1 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Leuconostoc mesenteroides | Q9L3Z9 | gene dsrS | - |
Leuconostoc mesenteroides NRRL B-512F | Q9L3Z9 | gene dsrS | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21 AI by nickel affinity chromatography | Leuconostoc mesenteroides |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
sucrose + [(1->6)-alpha-D-glucosyl]n | - |
Leuconostoc mesenteroides | D-fructose + [(1->6)-alpha-D-glucosyl]n+1 | - |
? | |
sucrose + [(1->6)-alpha-D-glucosyl]n | - |
Leuconostoc mesenteroides NRRL B-512F | D-fructose + [(1->6)-alpha-D-glucosyl]n+1 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Dsr S protein | - |
Leuconostoc mesenteroides |
DSR-S | - |
Leuconostoc mesenteroides |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Leuconostoc mesenteroides |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
38.2 | - |
sucrose | pH 5.2, 25°C, recombinant mutant D460M/H463Y/T464M/S512C | Leuconostoc mesenteroides | |
43.9 | - |
sucrose | pH 5.2, 25°C, recombinant mutant D460A/H463S/T464L | Leuconostoc mesenteroides | |
86 | - |
sucrose | pH 5.2, 25°C, recombinant mutant F353W | Leuconostoc mesenteroides | |
172.1 | - |
sucrose | pH 5.2, 25°C, recombinant mutant H463R/T464D/S512T | Leuconostoc mesenteroides | |
207.1 | - |
sucrose | pH 5.2, 25°C, recombinant mutant S512C | Leuconostoc mesenteroides | |
338.8 | - |
sucrose | pH 5.2, 25°C, recombinant mutant F353T | Leuconostoc mesenteroides | |
610.2 | - |
sucrose | pH 5.2, 25°C, recombinant mutant H463R/T464V/S512T | Leuconostoc mesenteroides |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5.2 | - |
assay at | Leuconostoc mesenteroides |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the glycoside-hydrolase family 70, sequence and structural analysis of GH70 enzymes, overview | Leuconostoc mesenteroides |
additional information | three-dimensional enzyme molecular structure modeling, structure comparisons, overview. Role of peptide 460DYVHT464 in enzyme DSR-S linkage specificity, together with residue S512 it contributes to defining the +2 subsite topology, which may be critical for the enzyme regiospecificity. Molecular docking of sucrose in the active site of enzyme mutant DSR-S vardel DELTA4N | Leuconostoc mesenteroides |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.5 | - |
sucrose | pH 5.2, 25°C, recombinant mutant D460M/H463Y/T464M/S512C | Leuconostoc mesenteroides | |
2.9 | - |
sucrose | pH 5.2, 25°C, recombinant mutant F353T | Leuconostoc mesenteroides | |
8.3 | - |
sucrose | pH 5.2, 25°C, recombinant mutant D460A/H463S/T464L | Leuconostoc mesenteroides | |
11.1 | - |
sucrose | pH 5.2, 25°C, recombinant mutant H463R/T464D/S512T | Leuconostoc mesenteroides | |
53.1 | - |
sucrose | pH 5.2, 25°C, recombinant mutant S512C | Leuconostoc mesenteroides | |
57.3 | - |
sucrose | pH 5.2, 25°C, recombinant mutant F353W | Leuconostoc mesenteroides | |
88.4 | - |
sucrose | pH 5.2, 25°C, recombinant mutant H463R/T464V/S512T | Leuconostoc mesenteroides |