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Literature summary extracted from

  • Dai, S.; Yao, Q.; Yu, G.; Liu, S.; Yun, J.; Xiao, X.; Deng, Z.; Li, H.
    Biochemical characterization of a novel bacterial laccase and improvement of its efficiency by directed evolution on dye degradation (2021), Front. Microbiol., 12, 633004 .
    View publication on PubMedView publication on EuropePMC

Application

EC Number Application Comment Organism
1.10.3.2 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(Commentary)

EC Number Cloned (Comment) Organism
1.10.3.2 transformed into Escherochia coli BL21(DE3) uncultured bacterium

Protein Variants

EC Number Protein Variants Comment Organism
1.10.3.2 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.10.3.2 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
1.10.3.2 0.21
-
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) pH and temperature not specified in the publication, wild-type enzyme uncultured bacterium
1.10.3.2 0.56
-
2,6-dimethoxyphenol pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V uncultured bacterium
1.10.3.2 0.654
-
2,6-dimethoxyphenol pH and temperature not specified in the publication, wild-type enzyme uncultured bacterium
1.10.3.2 4.3
-
guaiacol pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V uncultured bacterium
1.10.3.2 4.9
-
guaiacol pH and temperature not specified in the publication, wild-type enzyme uncultured bacterium
1.10.3.2 5.6
-
1-naphthol pH and temperature not specified in the publication, wild-type enzyme uncultured bacterium
1.10.3.2 6.04
-
1-naphthol pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V uncultured bacterium

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.10.3.2 copper copper-containing enzyme uncultured bacterium

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.10.3.2 68000
-
SDS-PAGE uncultured bacterium

Organism

EC Number Organism UniProt Comment Textmining
1.10.3.2 uncultured bacterium E1ACR6
-
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.10.3.2 0.089
-
substrate: 1-naphthol, pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V uncultured bacterium
1.10.3.2 0.13
-
substrate: 1-naphthol, pH and temperature not specified in the publication, wild-type enzyme uncultured bacterium
1.10.3.2 0.64
-
substrate: guaiacol, pH and temperature not specified in the publication, wild-type enzyme uncultured bacterium
1.10.3.2 1.12
-
substrate: 2,6-dimethoxyphenol, pH and temperature not specified in the publication, wild-type enzyme uncultured bacterium
1.10.3.2 1.37
-
substrate: guaiacol, pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V uncultured bacterium
1.10.3.2 2.67
-
substrate: 2,6-dimethoxyphenol, pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V uncultured bacterium
1.10.3.2 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
1.10.3.2 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 and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.10.3.2 1-naphthol + O2
-
uncultured bacterium ?
-
?
1.10.3.2 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + O2
-
uncultured bacterium ?
-
?
1.10.3.2 2,6-dimethoxyphenol + O2
-
uncultured bacterium ?
-
?
1.10.3.2 guaiacol + O2
-
uncultured bacterium ?
-
?
1.10.3.2 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

EC Number Subunits Comment Organism
1.10.3.2 ? x * 38000, SDS-PAGE uncultured bacterium

Synonyms

EC Number Synonyms Comment Organism
1.10.3.2 rlac1338
-
uncultured bacterium

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.10.3.2 55
-
wild-type enzyme uncultured bacterium
1.10.3.2 60
-
mutant enzyme V281A/P309L/S318G/D232V uncultured bacterium

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.10.3.2 35 65 35°C: about 65% of maximal activity, 65°C: about 50% of maximal activity, wild-type enzyme uncultured bacterium
1.10.3.2 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 [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.10.3.2 55
-
4 h, the relative enzyme activity is still over 50%, mutant enzyme V281A/P309L/S318G/D232V uncultured bacterium

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.10.3.2 1.34
-
guaiacol pH and temperature not specified in the publication, wild-type enzyme uncultured bacterium
1.10.3.2 1.57
-
guaiacol pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V uncultured bacterium
1.10.3.2 3.56
-
1-naphthol pH and temperature not specified in the publication, wild-type enzyme uncultured bacterium
1.10.3.2 3.89
-
1-naphthol pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V uncultured bacterium
1.10.3.2 9.43
-
2,6-dimethoxyphenol pH and temperature not specified in the publication, wild-type enzyme uncultured bacterium
1.10.3.2 11.23
-
2,6-dimethoxyphenol pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V uncultured bacterium
1.10.3.2 34.39
-
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) pH and temperature not specified in the publication, wild-type enzyme uncultured bacterium
1.10.3.2 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

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.10.3.2 6
-
wild-type enzyme uncultured bacterium
1.10.3.2 6.5
-
mutant enzyme V281A/P309L/S318G/D232V uncultured bacterium

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.10.3.2 4 8 pH 4.0: about 85% of maximal activity, pH 8.0: about 50% of maximal activity, wild-type enzyme uncultured bacterium
1.10.3.2 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

pI Value

EC Number Organism Comment pI Value Maximum pI Value
1.10.3.2 uncultured bacterium
-
-
5.05

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.10.3.2 0.27
-
guaiacol pH and temperature not specified in the publication, wild-type enzyme uncultured bacterium
1.10.3.2 0.37
-
guaiacol pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V uncultured bacterium
1.10.3.2 0.64
-
1-naphthol pH and temperature not specified in the publication, wild-type enzyme uncultured bacterium
1.10.3.2 0.64
-
1-naphthol pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V uncultured bacterium
1.10.3.2 14.4
-
2,6-dimethoxyphenol pH and temperature not specified in the publication, wild-type enzyme uncultured bacterium
1.10.3.2 20
-
2,6-dimethoxyphenol pH and temperature not specified in the publication, mutant enzyme V281A/P309L/S318G/D232V uncultured bacterium
1.10.3.2 160
-
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) pH and temperature not specified in the publication, wild-type enzyme uncultured bacterium
1.10.3.2 620
-
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