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 | 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 | 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 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 | Comment | Organism | Structure |
---|---|---|---|
Cu2+ | multicopper enzyme, the T1 copper centre clearly plays a key role | Bacillus subtilis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus subtilis | - |
strains AH3517, AH3522, and LOM401 | - |
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 | Comment | Organism |
---|---|---|
More | structure analysis of wild-type and mutant enzymes, crystal structure | Bacillus subtilis |
Synonyms | Comment | Organism |
---|---|---|
CotA laccase | - |
Bacillus subtilis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Bacillus subtilis |
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 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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.6 | - |
assay at | Bacillus subtilis |