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Literature summary for 1.10.3.2 extracted from
- Engineered Bacillus pumilus laccase-like multi-copper oxidase for enhanced oxidation of the lignin model compound guaiacol (2017), Protein Eng. Des. Sel., 30, 449-453 .
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| L386Q/G417I | variant L2 harbours the insertion of two amino acids (Ser and Pro) after the N-terminal methionine, in addition to the L386Q/G417I substitutions. The mutant enzyme exhibits 9-fold higher guaiacol oxidation rates in the lysate compared to the strain expressing the wild-type enzyme | Bacillus pumilus |
| L386Q/G417I/V482G | variant L9 harbours the insertion of two amino acids (Ser and Pro) after the N-terminal methionine, in addition to the L386Q/G417I/V482G substitutions. The mutant enzyme exhibits 14-fold higher guaiacol oxidation rates in the lysate compared to the strain expressing the wild-type enzyme. The decline in relative activity from the maximum at pH 4 towards more alkaline pH values is sharper for the mutant enzyme compared to the wild-type with 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid). The mutant enzyme has an increased activity towards the lignin-like model compound guaiacol while retaining the very good thermostability and activity at neutral-to-basic pH of the wild-type enzyme | Bacillus pumilus |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.11 | - |
2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 21-23°C, pH not specified in the publication, wild-type enzyme | Bacillus pumilus |  |
| 0.24 | - |
guaiacol | 22°C, pH 7.0, mutant enzyme L386Q/G417I (variant L2 harbours the insertion of two amino acids (Ser and Pro) after the N-terminal methionine, in addition to the L386Q/G417I substitutions) | Bacillus pumilus | |
| 0.41 | - |
2,6-dimethoxyphenol | 21-23°C, pH not specified in the publication, wild-type enzyme | Bacillus pumilus | |
| 1.05 | - |
guaiacol | 22°C, pH 7.0, wild-type enzyme | Bacillus pumilus | |
| 1.17 | - |
2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 21-23°C, pH not specified in the publication, mutant enzyme L386Q/G417I (variant L2 harbours the insertion of two amino acids (Ser and Pro) after the N-terminal methionine, in addition to the L386Q/G417I substitutions) | Bacillus pumilus | |
| 1.39 | - |
guaiacol | 22°C, pH 7.0, mutant enzyme L386Q/G417I/V482G (variant L9 harbours the insertion of two amino acids (Ser and Pro) after the N-terminal methionine, in addition to the L386Q/G417I/V482G substitutions) | Bacillus pumilus | |
| 1.83 | - |
2,6-dimethoxyphenol | 21-23°C, pH not specified in the publication, mutant enzyme L386Q/G417I/V482G (variant L9 harbours the insertion of two amino acids (Ser and Pro) after the N-terminal methionine, in addition to the L386Q/G417I/V482G substitutions) | Bacillus pumilus | |
| 2.06 | - |
2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 21-23°C, pH not specified in the publication, mutant enzyme L386Q/G417I/V482G 2.06 (variant L9 harbours the insertion of two amino acids (Ser and Pro) after the N-terminal methionine, in addition to the L386Q/G417I/V482G substitutions) | Bacillus pumilus | |
| 2.16 | - |
2,6-dimethoxyphenol | 21-23°C, pH not specified in the publication, mutant enzyme L386Q/G417I (variant L2 harbours the insertion of two amino acids (Ser and Pro) after the N-terminal methionine, in addition to the L386Q/G417I substitutions) | Bacillus pumilus | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Bacillus pumilus | K4PL78 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| wild-type and mutant enzymes | Bacillus pumilus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) + O2 | - |
Bacillus pumilus | ? | - |
? | |
| 2,6-dimethoxyphenol + O2 | - |
Bacillus pumilus | ? | - |
? | |
| guaiacol + O2 | - |
Bacillus pumilus | ? | - |
? | |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 70 | - |
wild-type and mutant enzymes retain more than 50% activity on both 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) and guaiacol upon incubation at 70 or 80°C for 2h | Bacillus pumilus |
| 80 | - |
wild-type and mutant enzymes retain more than 50% activity on both 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) and guaiacol upon incubation at 70 or 80°C for 2h | Bacillus pumilus |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 11.9 | - |
guaiacol | 22°C, pH 7.0, wild-type enzyme | Bacillus pumilus | |
| 15.7 | - |
2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 21-23°C, pH not specified in the publication, mutant enzyme L386Q/G417I/V482G (variant L9 harbours the insertion of two amino acids (Ser and Pro) after the N-terminal methionine, in addition to the L386Q/G417I/V482G substitutions) | Bacillus pumilus | |
| 23.2 | - |
2,6-dimethoxyphenol | 21-23°C, pH not specified in the publication, wild-type enzyme | Bacillus pumilus | |
| 38.8 | - |
2,6-dimethoxyphenol | 21-23°C, pH not specified in the publication, mutant enzyme L386Q/G417I/V482G (variant L9 harbours the insertion of two amino acids (Ser and Pro) after the N-terminal methionine, in addition to the L386Q/G417I/V482G substitutions) | Bacillus pumilus | |
| 39.5 | - |
guaiacol | 22°C, pH 7.0, mutant enzyme L386Q/G417I/V482G (variant L9 harbours the insertion of two amino acids (Ser and Pro) after the N-terminal methionine, in addition to the L386Q/G417I/V482G substitutions) | Bacillus pumilus | |
| 42.2 | - |
2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 21-23°C, pH not specified in the publication, wild-type enzyme | Bacillus pumilus | |
| 64.9 | - |
guaiacol | 22°C, pH 7.0, mutant enzyme L386Q/G417I (variant L2 harbours the insertion of two amino acids (Ser and Pro) after the N-terminal methionine, in addition to the L386Q/G417I substitutions) | Bacillus pumilus | |
| 69.7 | - |
2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 21-23°C, pH not specified in the publication, mutant enzyme L386Q/G417I (variant L2 harbours the insertion of two amino acids (Ser and Pro) after the N-terminal methionine, in addition to the L386Q/G417I substitutions) | Bacillus pumilus | |
| 80.4 | - |
2,6-dimethoxyphenol | 21-23°C, pH not specified in the publication, mutant enzyme L386Q/G417I (variant L2 harbours the insertion of two amino acids (Ser and Pro) after the N-terminal methionine, in addition to the L386Q/G417I substitutions) | Bacillus pumilus | |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 7.61 | - |
2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 21-23°C, pH not specified in the publication, mutant enzyme L386Q/G417I/V482G (variant L9 harbours the insertion of two amino acids (Ser and Pro) after the N-terminal methionine, in addition to the L386Q/G417I/V482G substitutions) | Bacillus pumilus | |
| 11.3 | - |
guaiacol | 22°C, pH 7.0, wild-type enzyme | Bacillus pumilus | |
| 21.2 | - |
2,6-dimethoxyphenol | 21-23°C, pH not specified in the publication, mutant enzyme L386Q/G417I/V482G (variant L9 harbours the insertion of two amino acids (Ser and Pro) after the N-terminal methionine, in addition to the L386Q/G417I/V482G substitutions) | Bacillus pumilus | |
| 28.5 | - |
guaiacol | 22°C, pH 7.0, mutant enzyme L386Q/G417I/V482G (variant L9 harbours the insertion of two amino acids (Ser and Pro) after the N-terminal methionine, in addition to the L386Q/G417I/V482G substitutions) | Bacillus pumilus | |
| 37.2 | - |
2,6-dimethoxyphenol | 21-23°C, pH not specified in the publication, mutant enzyme L386Q/G417I (variant L2 harbours the insertion of two amino acids (Ser and Pro) after the N-terminal methionine, in addition to the L386Q/G417I substitutions) | Bacillus pumilus | |
| 57 | - |
2,6-dimethoxyphenol | 21-23°C, pH not specified in the publication, wild-type enzyme | Bacillus pumilus | |
| 59.6 | - |
2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 21-23°C, pH not specified in the publication, mutant enzyme L386Q/G417I (variant L2 harbours the insertion of two amino acids (Ser and Pro) after the N-terminal methionine, in addition to the L386Q/G417I substitutions) | Bacillus pumilus | |
| 271 | - |
guaiacol | 22°C, pH 7.0, mutant enzyme L386Q/G417I (variant L2 harbours the insertion of two amino acids (Ser and Pro) after the N-terminal methionine, in addition to the L386Q/G417I substitutions) | Bacillus pumilus | |
| 383 | - |
2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 21-23°C, pH not specified in the publication, wild-type enzyme | Bacillus pumilus | |
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