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Literature summary for 2.4.1.10 extracted from

  • Mardo, K.; Visnapuu, T.; Gromkova, M.; Aasamets, A.; Viigand, K.; Vija, H.; Alamaee, T.
    High-throughput assay of levansucrase variants in search of feasible catalysts for the synthesis of fructooligosaccharides and levan (2014), Molecules, 19, 8434-8455.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.4.1.10

Application

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(Commentary)

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

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

Localization Comment Organism GeneOntology No. Textmining
extracellular
-
 Pseudomonas syringae pv. tomato
-
-

Natural Substrates/ Products (Substrates)

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

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 [micromol/min/mg]

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 and Products (Substrate)

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

Subunits Comment Organism
More three-dimensional structure comparison, overview Pseudomonas syringae pv. tomato

Synonyms

Synonyms Comment Organism
Lsc-3
-
 Pseudomonas syringae pv. tomato
Lsc3
-
 Pseudomonas syringae pv. tomato

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Pseudomonas syringae pv. tomato

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
 assay at Pseudomonas syringae pv. tomato

General Information

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