Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.10.3.2 extracted from

  • Durao, P.; Bento, I.; Fernandes, A.T.; Melo, E.P.; Lindley, P.F.; Martins, L.O.
    Perturbations of the T1 copper site in the CotA laccase from Bacillus subtilis: structural, biochemical, enzymatic and stability studies (2006), J. Biol. Inorg. Chem., 11, 514-526.
    View publication on PubMed
Search for more literature for 1.10.3.2

Crystallization (Commentary)

Crystallization (Comment) Organism
crystals of mutants M502L and M502F are obtained from a crystallization solution containing 12% 2-propanol, 12% of PEG 4000, 0.1 M sodium citrate, pH 5.5, and a protein concentration of about 5 mg/mL, vapour diffusion method, 2 days at room temperature, X-ray diffraction structure determination and analysis at 2.05-2.3 A resolution Bacillus subtilis

Protein Variants

Protein Variants Comment Organism
M502F site-directed mutagenesis, the mutation of the weak so-called axial ligand of the T1 copper site leads to an increase in the redox potential by approximately 100 mV relative to that of the wild-type enzyme, the mutant shows 10% and 0.15-0.05% activity for the non-phenolic substrates and for the phenolic substrates, respectively, compared with the wild-type enzyme Bacillus subtilis
M502L site-directed mutagenesis, the mutation of the weak so-called axial ligand of the T1 copper site leads to an increase in the redox potential by approximately 100 mV relative to that of the wild-type enzyme, the mutant exhibits a twofold to fourfold decrease in the kcat Bacillus subtilis

General Stability

General Stability Organism
unfolding of tertiary structure is a twostate process in the wild-type enzyme, displaying a midpoint at a guanidinium hydrochloride concentration of 4.6 M, unfolding for both mutant enzymes is clearly not a two-state process, overview Bacillus subtilis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information thermodynamics of wild-type and mutant enzymes, overview Bacillus subtilis
0.008
-
 syringaldazine pH 7.6, 37°C, mutant M502F Bacillus subtilis
0.009
-
 syringaldazine pH 7.6, 37°C, mutant M502L Bacillus subtilis
0.01
-
 syringaldazine pH 7.6, 37°C, wild-type enzyme Bacillus subtilis
0.027
-
 K4(FeCN6) pH 7.6, 37°C, mutant M502L Bacillus subtilis
0.035
-
 2,6-dimethoxyphenol pH 7.6, 37°C, mutant M502F Bacillus subtilis
0.049
-
 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) pH 7.6, 37°C, mutant M502F Bacillus subtilis
0.06
-
 2,6-dimethoxyphenol pH 7.6, 37°C, wild-type enzyme Bacillus subtilis
0.067
-
 K4(FeCN6) pH 7.6, 37°C, mutant M502F Bacillus subtilis
0.069
-
 K4(FeCN6) pH 7.6, 37°C, wild-type enzyme Bacillus subtilis
0.087
-
 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) pH 7.6, 37°C, wild-type enzyme Bacillus subtilis
0.089
-
 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) pH 7.6, 37°C, mutant M502L Bacillus subtilis
0.145
-
 2,6-dimethoxyphenol pH 7.6, 37°C, mutant M502L Bacillus subtilis

Metals/Ions

Metals/Ions Comment Organism Structure
Cu2+ multicopper enzyme, the T1 copper centre clearly plays a key role Bacillus subtilis

Organism

Organism UniProt Comment Textmining
Bacillus subtilis
-
 strains AH3517, AH3522, and LOM401
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) + O2
-
 Bacillus subtilis ?
-
 ?
2,6-dimethoxyphenol + O2
-
 Bacillus subtilis 3,3',5,5'-tetramethoxy-4-diphenoquinone + H2O
-
 ?
K4(FeCN6) + O2
-
 Bacillus subtilis ?
-
 ?
syringaldazine + O2
-
 Bacillus subtilis ?
-
 ?

Subunits

Subunits Comment Organism
More structure analysis of wild-type and mutant enzymes, crystal structure Bacillus subtilis

Synonyms

Synonyms Comment Organism
CotA laccase
-
 Bacillus subtilis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Bacillus subtilis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
-
 T1 copper depletion is a key event in inactivation and a determinant of the thermodynamic stability, thermodynamic stability of wild-type and mutant proteins, overview Bacillus subtilis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.01
-
 2,6-dimethoxyphenol pH 7.6, 37°C, mutant M502F Bacillus subtilis
0.01
-
 syringaldazine pH 7.6, 37°C, mutant M502F Bacillus subtilis
0.26
-
 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) pH 7.6, 37°C, mutant M502F Bacillus subtilis
1.55
-
 2,6-dimethoxyphenol pH 7.6, 37°C, mutant M502L Bacillus subtilis
5.5
-
 K4(FeCN6) pH 7.6, 37°C, mutant M502F Bacillus subtilis
6.65
-
 2,6-dimethoxyphenol pH 7.6, 37°C, wild-type enzyme Bacillus subtilis
7.4
-
 syringaldazine pH 7.6, 37°C, mutant M502L Bacillus subtilis
10.6
-
 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) pH 7.6, 37°C, mutant M502L Bacillus subtilis
18.4
-
 syringaldazine pH 7.6, 37°C, wild-type enzyme Bacillus subtilis
20.9
-
 K4(FeCN6) pH 7.6, 37°C, mutant M502L Bacillus subtilis
22.4
-
 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) pH 7.6, 37°C, wild-type enzyme Bacillus subtilis
54.5
-
 K4(FeCN6) pH 7.6, 37°C, wild-type enzyme Bacillus subtilis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.6
-
 assay at Bacillus subtilis