Application | Comment | Organism |
---|---|---|
synthesis | levansucrase Lsc3 of Pseudomonas syringae pv. tomato has very high catalytic activity and stability making it a promising biotechnological catalyst for FOS and levan synthesis | Pseudomonas syringae pv. tomato |
Cloned (Comment) | Organism |
---|---|
gene lsc3, sequence comparison, recombinant expression of N-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Pseudomonas syringae pv. tomato |
Protein Variants | Comment | Organism |
---|---|---|
D219A | site-directed mutagenesis, inactive mutant | Pseudomonas syringae pv. tomato |
D225A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Pseudomonas syringae pv. tomato |
D225N | site-directed mutagenesis, the mutant shows enhanced activity compared to the wild-type enzyme | Pseudomonas syringae pv. tomato |
D300A | site-directed mutagenesis, the mutant shows slightly enhanced activity compared to the wild-type enzyme | Pseudomonas syringae pv. tomato |
D333A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Pseudomonas syringae pv. tomato |
D333N | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Pseudomonas syringae pv. tomato |
D62A | site-directed mutagenesis, inactive mutant | Pseudomonas syringae pv. tomato |
E211Q | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Pseudomonas syringae pv. tomato |
E236Q | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Pseudomonas syringae pv. tomato |
E303A | site-directed mutagenesis, inactive mutant | Pseudomonas syringae pv. tomato |
E303Q | site-directed mutagenesis, inactive mutant | Pseudomonas syringae pv. tomato |
H113A | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Pseudomonas syringae pv. tomato |
H113Q | site-directed mutagenesis, inactive mutant | Pseudomonas syringae pv. tomato |
H306A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Pseudomonas syringae pv. tomato |
H321K | site-directed mutagenesis, almost inactive mutant | Pseudomonas syringae pv. tomato |
H321L | site-directed mutagenesis, almost inactive mutant | Pseudomonas syringae pv. tomato |
H321R | site-directed mutagenesis, almost inactive mutant | Pseudomonas syringae pv. tomato |
H321R | site-directed mutagenesis, significantly decreased polymerizing ability compared to the wild-type | Pseudomonas syringae pv. tomato |
H321S | site-directed mutagenesis, almost inactive mutant | Pseudomonas syringae pv. tomato |
L66A | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Pseudomonas syringae pv. tomato |
additional information | Vmax values for sucrose splitting of mutants, structure-function analysis of enzyme mutants, overview | Pseudomonas syringae pv. tomato |
P220A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Pseudomonas syringae pv. tomato |
Q301A | site-directed mutagenesis, significantly decreased polymerizing ability compared to the wild-type | Pseudomonas syringae pv. tomato |
Q301E | site-directed mutagenesis, the mutant shows slightly enhanced activity compared to the wild-type enzyme | Pseudomonas syringae pv. tomato |
R304A | site-directed mutagenesis, inactive mutant | Pseudomonas syringae pv. tomato |
R304C | site-directed mutagenesis, inactive mutant | Pseudomonas syringae pv. tomato |
T302M | site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme | Pseudomonas syringae pv. tomato |
T302P | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Pseudomonas syringae pv. tomato |
T302P | site-directed mutagenesis, significantly decreased polymerizing ability compared to the wild-type | Pseudomonas syringae pv. tomato |
W109A | site-directed mutagenesis, almost inactive mutant | Pseudomonas syringae pv. tomato |
W61A | site-directed mutagenesis, inactive mutant | Pseudomonas syringae pv. tomato |
W61N | site-directed mutagenesis, inactive mutant | Pseudomonas syringae pv. tomato |
W80R | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Pseudomonas syringae pv. tomato |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
extracellular | - |
Pseudomonas syringae pv. tomato | - |
- |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
sucrose + alpha-D-glucosyl-(1->2)-[(2->6)-alpha-D-fructosyl]n | Pseudomonas syringae pv. tomato | - |
D-glucose + alpha-D-glucosyl-(1->2)-[(2->6)-alpha-D-fructosyl]n+1 | - |
? | |
sucrose + alpha-D-glucosyl-(1->2)-[(2->6)-alpha-D-fructosyl]n | Pseudomonas syringae pv. tomato DC3000 | - |
D-glucose + alpha-D-glucosyl-(1->2)-[(2->6)-alpha-D-fructosyl]n+1 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas syringae pv. tomato | Q88BN6 | pv. tomato, gene lsc3 | - |
Pseudomonas syringae pv. tomato DC3000 | Q88BN6 | pv. tomato, gene lsc3 | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
transfructosylation activity of wild-type and mutant enzymes (permeabilized cells/purified protein), overview | Pseudomonas syringae pv. tomato |
461.6 | - |
purified recombinant wild-type enzyme, pH 6.0, 37°C | Pseudomonas syringae pv. tomato |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | assay of the growth phenotype of levansucrase-expressing bacteria on agar plate containing sucrose. Only wild-type Lsc3 produces levan giving a mucoid phenotype to the streaks of respective transformant. Quantitative evaluation of the sucrose-splitting (total) levansucrase activity on microplates | Pseudomonas syringae pv. tomato | ? | - |
? | |
additional information | assay of the growth phenotype of levansucrase-expressing bacteria on agar plate containing sucrose. Only wild-type Lsc3 produces levan giving a mucoid phenotype to the streaks of respective transformant. Quantitative evaluation of the sucrose-splitting (total) levansucrase activity on microplates | Pseudomonas syringae pv. tomato DC3000 | ? | - |
? | |
sucrose + alpha-D-glucosyl-(1->2)-[(2->6)-alpha-D-fructosyl]n | - |
Pseudomonas syringae pv. tomato | D-glucose + alpha-D-glucosyl-(1->2)-[(2->6)-alpha-D-fructosyl]n+1 | - |
? | |
sucrose + alpha-D-glucosyl-(1->2)-[(2->6)-alpha-D-fructosyl]n | - |
Pseudomonas syringae pv. tomato DC3000 | D-glucose + alpha-D-glucosyl-(1->2)-[(2->6)-alpha-D-fructosyl]n+1 | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | three-dimensional structure comparison, overview | Pseudomonas syringae pv. tomato |
Synonyms | Comment | Organism |
---|---|---|
Lsc-3 | - |
Pseudomonas syringae pv. tomato |
Lsc3 | - |
Pseudomonas syringae pv. tomato |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Pseudomonas syringae pv. tomato |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
assay at | Pseudomonas syringae pv. tomato |
General Information | Comment | Organism |
---|---|---|
metabolism | bacterial levansucrases polymerize fructose residues of sucrose to beta-2,6 linked fructans-fructooligosaccharides (FOS) and levan | Pseudomonas syringae pv. tomato |
additional information | the enzyme has very high catalytic activity and stability. Key importance of residues Trp109, His113, Glu146 and Glu236 for the catalysis of Lsc3, the catalytic triad is formed by D62, D219, and E303 | Pseudomonas syringae pv. tomato |