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Literature summary for 2.4.1.5 extracted from

  • Nam, S.H.; Ko, E.A.; Jang, S.S.; Kim, d.o..W.; Kim, S.Y.; Hwang, D.S.; Kim, D.
    Maximization of dextransucrase activity expressed in E. coli by mutation and its functional characterization (2008), Biotechnol. Lett., 30, 135-143.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene DSRB742, expression of wild-type and DRN1-DRN4 mutant enzymes in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha Leuconostoc mesenteroides

Protein Variants

Protein Variants Comment Organism
F196S site-directed mutagenesis, the mutant DRN1 shows a higher expression level in Escherichia coli and increased activity compared to DSRB742 Leuconostoc mesenteroides
K395T site-directed mutagenesis, the mutant DRN3 shows a higher expression level in Escherichia coli and increased activity compared to DSRB742 Leuconostoc mesenteroides
P980T site-directed mutagenesis, the mutant DRN4 shows a lower expression level in Escherichia coli compared to DSRB742 Leuconostoc mesenteroides
Y346N site-directed mutagenesis, the mutant DRN2 shows a lower expression level in Escherichia coli compared to DSRB742 Leuconostoc mesenteroides

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
18
-
sucrose pH 5.2, 28°C, recombinant mutant DRN1 Leuconostoc mesenteroides
48
-
sucrose pH 5.2, 28°C, recombinant mutant DRN3 Leuconostoc mesenteroides
79
-
sucrose pH 5.2, 28°C, recombinant mutant DRN4 Leuconostoc mesenteroides
83
-
sucrose pH 5.2, 28°C, recombinant mutant DRN2 Leuconostoc mesenteroides
88
-
sucrose pH 5.2, 28°C, recombinant wild-type enzyme Leuconostoc mesenteroides

Organism

Organism UniProt Comment Textmining
Leuconostoc mesenteroides
-
gene DSRB742
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and DRN1-DRN4 mutant enzymes from Escherichia coli by nickel affinity chromatography Leuconostoc mesenteroides

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.012
-
purified recombinant mutant DRN4 Leuconostoc mesenteroides
0.027
-
purified recombinant mutant DRN2 Leuconostoc mesenteroides
0.037
-
purified recombinant wild-type DSRB742 Leuconostoc mesenteroides
0.185
-
purified recombinant mutant DRN3 Leuconostoc mesenteroides
0.287
-
purified recombinant mutant DRN1 Leuconostoc mesenteroides

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
sucrose + gentobiose glucose transfer from donor sucrose to acceptors releasing D-fructose, acceptor specificity of wild-type and mutant enzymes, overview Leuconostoc mesenteroides ?
-
?
sucrose + maltose glucose transfer from donor sucrose to acceptors releasing D-fructose, acceptor specificity of wild-type and mutant enzymes, overview Leuconostoc mesenteroides ?
-
?
sucrose + salicin glucose transfer from donor sucrose to acceptors releasing D-fructose, acceptor specificity of wild-type and mutant enzymes, overview Leuconostoc mesenteroides ?
-
?

Synonyms

Synonyms Comment Organism
DSRB742
-
Leuconostoc mesenteroides

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
28
-
assay at Leuconostoc mesenteroides

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.4
-
sucrose pH 5.2, 28°C, recombinant mutant DRN4 Leuconostoc mesenteroides
0.6
-
sucrose pH 5.2, 28°C, recombinant mutant DRN2 Leuconostoc mesenteroides
1.8
-
sucrose pH 5.2, 28°C, recombinant wild-type enzyme Leuconostoc mesenteroides
2.4
-
sucrose pH 5.2, 28°C, recombinant mutant DRN1 Leuconostoc mesenteroides
2.8
-
sucrose pH 5.2, 28°C, recombinant mutant DRN3 Leuconostoc mesenteroides

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.2
-
assay at Leuconostoc mesenteroides