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Literature summary for 1.10.3.2 extracted from

  • Zhu, Y.; Zhan, J.; Zhang, Y.; Lin, Y.; Yang, X.
    The K428 residue from Thermus thermophilus SG0.5JP17-16 laccase plays the substantial role in substrate binding and oxidation (2021), J. Biomol. Struct. Dyn., 39, 1312-1320 .
    View publication on PubMed
Search for more literature for 1.10.3.2

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli BL21 Thermus thermophilus

Protein Variants

Protein Variants Comment Organism
K428E 1.3fold decrease in kcat/Km for the substrate guaiacol Thermus thermophilus
K428L 1.6fold decrease in kcat/Km for the substrate guaiacol. 70% decrease in activity of mutant enzyme after 4 h at 80°C. 30% decrease in activity of wild-type enzyme after 4 h at 80°C Thermus thermophilus
K428M 1.4fold increase in kcat/Km for the substrate guaiacol Thermus thermophilus
K428R 1.3fold decrease in kcat/Km for the substrate guaiacol Thermus thermophilus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.384
-
 guaiacol pH 6.0, 90°C, wild-type enzyme Thermus thermophilus
0.391
-
 guaiacol pH 6.0, 90°C, mutant enzyme K428M Thermus thermophilus
0.473
-
 guaiacol pH 6.0, 90°C, mutant enzyme K428R Thermus thermophilus
0.52
-
 guaiacol pH 6.0, 90°C, mutant enzyme K428E Thermus thermophilus
0.575
-
 guaiacol pH 6.0, 90°C, mutant enzyme K428L Thermus thermophilus

Organism

Organism UniProt Comment Textmining
Thermus thermophilus
-
-
-
Thermus thermophilus SG0.5JP17-16
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Thermus thermophilus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
guaiacol + O2
-
 Thermus thermophilus ?
-
 ?
guaiacol + O2
-
 Thermus thermophilus SG0.5JP17-16 ?
-
 ?

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
90
-
 wild-type and K428L, K428M, K428R and K428E mutant enzymes Thermus thermophilus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
80
-
 0-4 h, residual enzymatic activity decreases with the increase of the incubation time. The wild-type, K428M, K428E and K428R enzymes remain 70% of corresponding initial activity after incubated for 4 h. The K428L mutant is most affected, only 30% of the initial activity is retained after 4 h Thermus thermophilus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.7
-
 guaiacol pH 6.0, 90°C, mutant enzyme K428R Thermus thermophilus
5.8
-
 guaiacol pH 6.0, 90°C, mutant enzyme K428L Thermus thermophilus
6.1
-
 guaiacol pH 6.0, 90°C, wild-type enzyme Thermus thermophilus
6.2
-
 guaiacol pH 6.0, 90°C, mutant enzyme K428E Thermus thermophilus
8.4
-
 guaiacol pH 6.0, 90°C, mutant enzyme K428M Thermus thermophilus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
 wild-type and K428L, K428M, K428R and K428E mutant enzymes Thermus thermophilus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
10
-
 guaiacol pH 6.0, 90°C, mutant enzyme K428L Thermus thermophilus
12
-
 guaiacol pH 6.0, 90°C, mutant enzyme K428E Thermus thermophilus
12
-
 guaiacol pH 6.0, 90°C, mutant enzyme K428R Thermus thermophilus
16
-
 guaiacol pH 6.0, 90°C, wild-type enzyme Thermus thermophilus
22
-
 guaiacol pH 6.0, 90°C, mutant enzyme K428M Thermus thermophilus