Cloned (Comment) | Organism |
---|---|
Ery4 genetic structure, recombinant expression of wild-type and mutant enzymes in Saccharomyces cerevisiae | Pleurotus eryngii |
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 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 | UniProt | Comment | Textmining |
---|---|---|---|
Pleurotus eryngii | B0JDP9 | gene ERY4 | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Saccharomyces cerevisiae by anion exchange chromatography and gel filtration | Pleurotus eryngii |
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 | Comment | Organism |
---|---|---|
More | enzyme structure modeling, structure comparions, overview | Pleurotus eryngii |
Synonyms | Comment | Organism |
---|---|---|
Ery4 | - |
Pleurotus eryngii |
Ery4 laccase | - |
Pleurotus eryngii |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Pleurotus eryngii |
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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
3 | 6.5 | assay at | Pleurotus eryngii |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
pH dependencies of wild-type and mutant enzymes, overview | Pleurotus eryngii |
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 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 |