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Literature summary for 1.10.3.2 extracted from

  • Bleve, G.; Lezzi, C.; Spagnolo, S.; Tasco, G.; Tufariello, M.; Casadio, R.; Mita, G.; Rampino, P.; Grieco, F.
    Role of the C-terminus of Pleurotus eryngii Ery4 laccase in determining enzyme structure, catalytic properties and stability (2013), Protein Eng. Des. Sel., 26, 1-13.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
Ery4 genetic structure, recombinant expression of wild-type and mutant enzymes in Saccharomyces cerevisiae Pleurotus eryngii

Protein Variants

Protein Variants Comment Organism
K532A site-directed mutagenesis, the mutant shows increased catalytic efficiency and altered substrate specificity compared to the wild-type enzyme Pleurotus eryngii
K532E site-directed mutagenesis, the mutant shows increased catalytic efficiency and altered substrate specificity compared to the wild-type enzyme Pleurotus eryngii
K532R site-directed mutagenesis, inactive mutant Pleurotus eryngii
additional information construction and generation of C-terminal truncation mutants lacking 1, 2, 5, 8, 11, 14 or 18 amino acid residues Pleurotus eryngii
P530A site-directed mutagenesis, the mutant shows increased catalytic efficiency and altered substrate specificity compared to the wild-type enzyme Pleurotus eryngii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetics of wild-type and mutant enzymes, overview Pleurotus eryngii
0.058
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 3.0, 25°C, mutant K532A Pleurotus eryngii
0.084
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 3.0, 25°C, mutant K532E Pleurotus eryngii
0.105
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 3.0, 25°C, mutant P530A Pleurotus eryngii
0.653
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 4.5, 25°C, mutant K532A Pleurotus eryngii
1.219
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 4.5, 25°C, mutant P530A Pleurotus eryngii
1.278
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 4.5, 25°C, mutant K532E Pleurotus eryngii
2.174
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 6.5, 25°C, mutant K532A Pleurotus eryngii
2.487
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 6.5, 25°C, mutant K532E Pleurotus eryngii
3.703
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 6.5, 25°C, mutant P530A Pleurotus eryngii

Organism

Organism UniProt Comment Textmining
Pleurotus eryngii B0JDP9 gene ERY4
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Saccharomyces cerevisiae by anion exchange chromatography and gel filtration Pleurotus eryngii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) + O2
-
Pleurotus eryngii ?
-
?
4 2,6-dimethoxyphenol + O2
-
Pleurotus eryngii ?
-
?
4 3,3',5,5'-tetramethylbenzidine + O2
-
Pleurotus eryngii ?
-
?
4 guaiacol + O2
-
Pleurotus eryngii ?
-
?
4 syringaldazine + O2
-
Pleurotus eryngii ?
-
?

Subunits

Subunits Comment Organism
More enzyme structure modeling, structure comparions, overview Pleurotus eryngii

Synonyms

Synonyms Comment Organism
Ery4
-
Pleurotus eryngii
Ery4 laccase
-
Pleurotus eryngii

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Pleurotus eryngii

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.26
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 6.5, 25°C, mutant K532A Pleurotus eryngii
4.58
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 6.5, 25°C, mutant P530A Pleurotus eryngii
5.51
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 6.5, 25°C, mutant K532E Pleurotus eryngii
60.6
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 4.5, 25°C, mutant P530A Pleurotus eryngii
70.02
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 4.5, 25°C, mutant K532A Pleurotus eryngii
70.6
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 3.0, 25°C, mutant P530A Pleurotus eryngii
74.67
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 4.5, 25°C, mutant K532E Pleurotus eryngii
97.85
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 3.0, 25°C, mutant K532E Pleurotus eryngii
100.7
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 3.0, 25°C, mutant K532A Pleurotus eryngii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
3 6.5 assay at Pleurotus eryngii

pH Range

pH Minimum pH Maximum Comment Organism
additional information
-
pH dependencies of wild-type and mutant enzymes, overview Pleurotus eryngii

General Information

General Information Comment Organism
additional information the last 18 amino acids in the C-terminal end of the Ery4 laccase play a critical role in enzyme activity, stability and kinetic and, in particular biochemical and structural data indicate that the K532 residue is fundamental for enzyme activation. Structure-function relationships of fungal laccases, overview Pleurotus eryngii

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.074
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 6.5, 25°C, mutant P530A Pleurotus eryngii
0.133
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 6.5, 25°C, mutant K532E Pleurotus eryngii
0.154
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 6.5, 25°C, mutant K532A Pleurotus eryngii
0.672
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 3.0, 25°C, mutant P530A Pleurotus eryngii
1.165
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 3.0, 25°C, mutant K532E Pleurotus eryngii
1.73
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 3.0, 25°C, mutant K532A Pleurotus eryngii
2.981
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 4.5, 25°C, mutant P530A Pleurotus eryngii
3.505
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 4.5, 25°C, mutant K532E Pleurotus eryngii
6.434
-
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) pH 4.5, 25°C, mutant K532A Pleurotus eryngii