Application | Comment | Organism |
---|---|---|
environmental protection | good application prospect in wastewater treatment and dye degradation. error-prone PCR is a feasible method to improve the degradation activity of laccase for environmental pollutants, which provide a basis for the application of laccase on dye degradation and other environmental pollutants | uncultured bacterium |
Cloned (Comment) | Organism |
---|---|
transformed into Escherochia coli BL21(DE3) | uncultured bacterium |
Protein Variants | Comment | Organism |
---|---|---|
V281A/P309L/S318G/D232V | the expression of the optimized mutant enzyme increases by 22% compared to the unoptimized enzyme and the optimal reaction temperature of the mutant enzyme is 5°C higher than that of the recombinant wild-type enzyme rlac1338, and the optimal pH increases by 0.5 units. The thermal stability and pH stability of the mutant enzyme lac2-9 are improved. 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) is the most suitable substrate for the recombinant enzyme and mutant enzyme. In addition, the Km of the mutant strain lac2-9 (76 mM) is significantly lower, but the kcat/Km (0.618 /s*M) is significantly higher, and the specific enzyme activity (79.8 U/mg) increases by 3.5 times compared with the recombinant laccase (22.8 U/mg). Compared to the rlac1338, the degradation rates with the simultaneous addition of Ca2+ and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) of mutant enzyme lac2-9 for acid violet 7, bromophenol blue and coomassie brilliant blue significantly improved by 8.3, 3.4 and 3.4 times | uncultured bacterium |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.076 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V | uncultured bacterium | |
0.21 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | pH and temperature not specified in the publication, wild-type enzyme | uncultured bacterium | |
0.56 | - |
2,6-dimethoxyphenol | pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V | uncultured bacterium | |
0.654 | - |
2,6-dimethoxyphenol | pH and temperature not specified in the publication, wild-type enzyme | uncultured bacterium | |
4.3 | - |
guaiacol | pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V | uncultured bacterium | |
4.9 | - |
guaiacol | pH and temperature not specified in the publication, wild-type enzyme | uncultured bacterium | |
5.6 | - |
1-naphthol | pH and temperature not specified in the publication, wild-type enzyme | uncultured bacterium | |
6.04 | - |
1-naphthol | pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V | uncultured bacterium |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
copper | copper-containing enzyme | uncultured bacterium |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
68000 | - |
SDS-PAGE | uncultured bacterium |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
uncultured bacterium | E1ACR6 | - |
- |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.089 | - |
substrate: 1-naphthol, pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V | uncultured bacterium |
0.13 | - |
substrate: 1-naphthol, pH and temperature not specified in the publication, wild-type enzyme | uncultured bacterium |
0.64 | - |
substrate: guaiacol, pH and temperature not specified in the publication, wild-type enzyme | uncultured bacterium |
1.12 | - |
substrate: 2,6-dimethoxyphenol, pH and temperature not specified in the publication, wild-type enzyme | uncultured bacterium |
1.37 | - |
substrate: guaiacol, pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V | uncultured bacterium |
2.67 | - |
substrate: 2,6-dimethoxyphenol, pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V | uncultured bacterium |
22.8 | - |
substrate: 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), pH and temperature not specified in the publication, wild-type enzyme | uncultured bacterium |
79.8 | - |
substrate: 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V | uncultured bacterium |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1-naphthol + O2 | - |
uncultured bacterium | ? | - |
? | |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + O2 | - |
uncultured bacterium | ? | - |
? | |
2,6-dimethoxyphenol + O2 | - |
uncultured bacterium | ? | - |
? | |
guaiacol + O2 | - |
uncultured bacterium | ? | - |
? | |
additional information | the use of rlac1338 and lac2-9 alone had no degradation effect on the industrial dyes [indigo, amaranth, bromophenol blue, acid violet 7, Congo red, coomassie brilliant blue (G250)], however, adding small molecular mediators Ca2+ and 2,2'-azino-bis(3-ethylbenzothiazo-line)-6-sulfonic acid at the same time can significantly improve the degradation ability | uncultured bacterium | ? | - |
- |
Subunits | Comment | Organism |
---|---|---|
? | x * 38000, SDS-PAGE | uncultured bacterium |
Synonyms | Comment | Organism |
---|---|---|
rlac1338 | - |
uncultured bacterium |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
55 | - |
wild-type enzyme | uncultured bacterium |
60 | - |
mutant enzyme V281A/P309L/S318G/D232V | uncultured bacterium |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
35 | 65 | 35°C: about 65% of maximal activity, 65°C: about 50% of maximal activity, wild-type enzyme | uncultured bacterium |
35 | 65 | 35°C: about 65% of maximal activity, 65°C: about 75% of maximal activity, mutant enzyme V281A/P309L/S318G/D232V | uncultured bacterium |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
55 | - |
4 h, the relative enzyme activity is still over 50%, mutant enzyme V281A/P309L/S318G/D232V | uncultured bacterium |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.34 | - |
guaiacol | pH and temperature not specified in the publication, wild-type enzyme | uncultured bacterium | |
1.57 | - |
guaiacol | pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V | uncultured bacterium | |
3.56 | - |
1-naphthol | pH and temperature not specified in the publication, wild-type enzyme | uncultured bacterium | |
3.89 | - |
1-naphthol | pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V | uncultured bacterium | |
9.43 | - |
2,6-dimethoxyphenol | pH and temperature not specified in the publication, wild-type enzyme | uncultured bacterium | |
11.23 | - |
2,6-dimethoxyphenol | pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V | uncultured bacterium | |
34.39 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | pH and temperature not specified in the publication, wild-type enzyme | uncultured bacterium | |
46.94 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V | uncultured bacterium |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
wild-type enzyme | uncultured bacterium |
6.5 | - |
mutant enzyme V281A/P309L/S318G/D232V | uncultured bacterium |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
4 | 8 | pH 4.0: about 85% of maximal activity, pH 8.0: about 50% of maximal activity, wild-type enzyme | uncultured bacterium |
4 | 8 | pH 4.0: about 90% of maximal activity, pH 8.0: about 50% of maximal activity, mutant enzyme V281A/P309L/S318G/D232V | uncultured bacterium |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
uncultured bacterium | - |
- |
5.05 |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.27 | - |
guaiacol | pH and temperature not specified in the publication, wild-type enzyme | uncultured bacterium | |
0.37 | - |
guaiacol | pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V | uncultured bacterium | |
0.64 | - |
1-naphthol | pH and temperature not specified in the publication, wild-type enzyme | uncultured bacterium | |
0.64 | - |
1-naphthol | pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V | uncultured bacterium | |
14.4 | - |
2,6-dimethoxyphenol | pH and temperature not specified in the publication, wild-type enzyme | uncultured bacterium | |
20 | - |
2,6-dimethoxyphenol | pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V | uncultured bacterium | |
160 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | pH and temperature not specified in the publication, wild-type enzyme | uncultured bacterium | |
620 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V | uncultured bacterium |