Cloned (Comment) | Organism |
---|---|
DNA and amino acid sequence determination and analysis, subcloning in Escherichia coli strain Top10, expression of wild-type and mutant enzymes in Saccharomyces cerevisiae strain W303-1A | Pleurotus ostreatus |
Protein Variants | Comment | Organism |
---|---|---|
D205R | site-directed mutagenesis, the mutation in a highly conserved region perturbs the structural local environment in POXA1b, leading to a large rearrangement of the enzyme structure. The mutant shows highly reduced activity compared to the wild-type enzyme and is inactive with substrate syringaldazine | Pleurotus ostreatus |
additional information | deletion of the C-terminal extension of 4 or 16 amino acids leads to truncated mutants which lose the high stability at pH 10, while they show an increased stability at pH 5.0, the thermostability is unaltered compared to the wild-type enzyme | Pleurotus ostreatus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | construction of enzymesubstrate complexes and molecular dynamics simulations, complex of mutant D205R with 2,5-xylidine, molecular dynamics simulations, overview | Pleurotus ostreatus | |
0.03 | - |
syringaldazine | pH 6.0, 22°C, recombinant truncated mutant POXA1bDELTA16 | Pleurotus ostreatus | |
0.03 | - |
syringaldazine | pH 6.0, 22°C, recombinant truncated mutant POXA1bDELTA4 | Pleurotus ostreatus | |
0.04 | - |
syringaldazine | pH 6.0, 22°C, recombinant wild-type enzyme | Pleurotus ostreatus | |
0.07 | - |
2,2'-azino-bis(3-ethyl-benzothiazoline-6-sulfonic acid) | pH 3.0, 22°C, recombinant wild-type enzyme | Pleurotus ostreatus | |
0.11 | - |
2,2'-azino-bis(3-ethyl-benzothiazoline-6-sulfonic acid) | pH 3.0, 22°C, recombinant truncated mutant POXA1bDELTA16 | Pleurotus ostreatus | |
0.11 | - |
2,2'-azino-bis(3-ethyl-benzothiazoline-6-sulfonic acid) | pH 3.0, 22°C, recombinant truncated mutant POXA1bDELTA4 | Pleurotus ostreatus | |
0.32 | - |
2,2'-azino-bis(3-ethyl-benzothiazoline-6-sulfonic acid) | pH 3.0, 22°C, recombinant truncated mutant POXA1bD205R | Pleurotus ostreatus | |
0.43 | - |
2,6-dimethoxyphenol | pH 5.3, 22°C, recombinant truncated mutant POXA1bDELTA16 | Pleurotus ostreatus | |
0.43 | - |
2,6-dimethoxyphenol | pH 5.3, 22°C, recombinant truncated mutant POXA1bDELTA4 | Pleurotus ostreatus | |
0.57 | - |
2,6-dimethoxyphenol | pH 5.3, 22°C, recombinant wild-type enzyme | Pleurotus ostreatus | |
1.26 | - |
2,6-dimethoxyphenol | pH 5.3, 22°C, recombinant truncated mutant POXA1bD205R | Pleurotus ostreatus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Cu2+ | laccases contain four copper ions distributed into three sites that, according to their spectroscopic properties, are classified as type 1 Cu, or blue copper centre, type 2 or normal copper centres, and type 3 or coupled binuclear copper centres, overview | Pleurotus ostreatus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pleurotus ostreatus | O60199 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes partially from Saccharomyces cerevisiae strain W303-1A | Pleurotus ostreatus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
3 | - |
partially purified recombinant truncated mutant POXA1bD205R, substrate 2,6-dimethoxyphenol | Pleurotus ostreatus |
12 | - |
partially purified recombinant truncated mutant POXA1bDELTA16, substrate syringaldazine | Pleurotus ostreatus |
15 | - |
partially purified recombinant wild-type enzyme, substrate syringaldazine | Pleurotus ostreatus |
17 | - |
partially purified recombinant truncated mutant POXA1bDELTA4, substrate syringaldazine | Pleurotus ostreatus |
33 | - |
partially purified recombinant truncated mutant POXA1bD205R, substrate 2,2'-azinobis(3-ethylbenzo-6-thiazolinesulfonic) acid | Pleurotus ostreatus |
33 | - |
partially purified recombinant truncated mutant POXA1bDELTA16, substrate 2,2'-azinobis(3-ethylbenzo-6-thiazolinesulfonic) acid | Pleurotus ostreatus |
100 | - |
partially purified recombinant truncated mutant POXA1bDELTA4, substrate 2,2'-azinobis(3-ethylbenzo-6-thiazolinesulfonic) acid | Pleurotus ostreatus |
129 | - |
partially purified recombinant truncated mutant POXA1bDELTA16, substrate 2,6-dimethoxyphenol | Pleurotus ostreatus |
155 | - |
partially purified recombinant wild-type enzyme, substrate 2,6-dimethoxyphenol | Pleurotus ostreatus |
162 | - |
partially purified recombinant wild-type enzyme, substrate 2,2'-azinobis(3-ethylbenzo-6-thiazolinesulfonic) acid | Pleurotus ostreatus |
189 | - |
partially purified recombinant truncated mutant POXA1bDELTA4, substrate 2,6-dimethoxyphenol | Pleurotus ostreatus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) + O2 | - |
Pleurotus ostreatus | ? | - |
? | |
2,6-dimethoxyphenol + O2 | - |
Pleurotus ostreatus | ? | - |
? | |
syringaldazine + O2 | - |
Pleurotus ostreatus | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | the POXA1b laccase possesses an unusual C-terminal extension of sixteen amino acids compared to other laccases | Pleurotus ostreatus |
Synonyms | Comment | Organism |
---|---|---|
POXA1b | - |
Pleurotus ostreatus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
22 | - |
assay at room tenperature | Pleurotus ostreatus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
3 | - |
assay at, substrate 2,2'-azinobis(3-ethylbenzo-6-thiazolinesulfonic) acid | Pleurotus ostreatus |
5.3 | - |
assay at, substrate 2,6-dimethoxyphenol | Pleurotus ostreatus |
6 | - |
assay at, substrate syringaldazine | Pleurotus ostreatus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
3 | 6.4 | pH profile with substrate 2,6-dimethoxyphenol | Pleurotus ostreatus |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
3 | 10 | stability of recombinant wild-type and mutant enzymes, overview | Pleurotus ostreatus |
10 | - |
half-life are 30 days | Pleurotus ostreatus |