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Literature summary for 1.10.3.2 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

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(Commentary)

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

Protein Variants

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 [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

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

Molecular Weight [Da]

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

Organism

Organism UniProt Comment Textmining
uncultured bacterium E1ACR6
-
-

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

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

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

Synonyms

Synonyms Comment Organism
rlac1338
-
uncultured bacterium

Temperature Optimum [°C]

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 Range [°C]

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 [°C]

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 [1/s]

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

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 Range

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

pI Value

Organism Comment pI Value Maximum pI Value
uncultured bacterium
-
-
5.05

kcat/KM [mM/s]

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