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Literature summary for 1.10.3.2 extracted from
- Heterologous expression, engineering and characterization of a novel laccase of Agrocybe pediades with promising properties as biocatalyst (2021), J. Fungi (Basel), 7, 359 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed in Saccharomyces cerevisiae using an improved signal peptide previously obtained and enzyme directed evolution | Agrocybe pediades |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| N398D | twice better activity at pH 6 and 1.2fold improvement at pH 3 | Agrocybe pediades |
| V159E | two-fold improvement in the laccase activity detected at pH 3 and 6. Lower thermotolerance than native enzyme | Agrocybe pediades |
| V159E/N398D | 1.4fold higher activity than mutant enzyme V159E and 2.6fold higher activity than mutant enzyme N398D with 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) as substrate. Lower thermotolerance than native enzyme | Agrocybe pediades |
| V159E/N398D/I453F/M454L | parental-like activity at both pH values. Lower thermotolerance than native enzyme | Agrocybe pediades |
| V159E/N398D/I453L/M454L | 1.3fold TAI at pH 3 and lower activity (0.8fold) than mutant enzyme V159E/N398D. Lower thermotolerance than native enzyme | Agrocybe pediades |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| acetone | mutant enzyme V159E/N398D | Agrocybe pediades |  |
| DMSO | mutant enzyme V159E/N398D | Agrocybe pediades | |
| EDTA | mutant enzyme V159E/N398D | Agrocybe pediades | |
| ethanol | mutant enzyme V159E/N398D | Agrocybe pediades | |
| NaCl | mutant enzyme V159E/N398D | Agrocybe pediades | |
| NaF | mutant enzyme V159E/N398D | Agrocybe pediades | |
| SDS | mutant enzyme V159E/N398D | Agrocybe pediades | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0045 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | pH 3, temperature not specified in the publication, mutant enzyme V159E | Agrocybe pediades | |
| 0.0046 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | pH 3, temperature not specified in the publication, mutant enzyme V159E/N398D | Agrocybe pediades | |
| 0.016 | - |
2,6-dimethoxyphenol | pH 5, temperature not specified in the publication, mutant enzyme V159E | Agrocybe pediades | |
| 0.08 | - |
2,6-dimethoxyphenol | pH 5, temperature not specified in the publication, mutant enzyme V159E/N398D | Agrocybe pediades | |
| 0.53 | - |
N,N-dimethyl-1,4-phenylenediamine | pH 4, temperature not specified in the publication, mutant enzyme V159E | Agrocybe pediades | |
| 1.7 | - |
N,N-dimethyl-1,4-phenylenediamine | pH 4, temperature not specified in the publication, mutant enzyme V159E/N398D | Agrocybe pediades | |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| extracellular | secreted by the fungus during lignocellulose degradation | Agrocybe pediades | - |
- |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| copper | multicopper oxidase | Agrocybe pediades | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Agrocybe pediades | - |
- |
- |
| Agrocybe pediades AH40210 | - |
- |
- |
Posttranslational Modification
| Posttranslational Modification | Comment | Organism |
|---|---|---|
| glycoprotein | existence of three N-glycosylation sites in the laccase and their distinct effect on the secretion or catalytic activity of the enzyme is demonstrated | Agrocybe pediades |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
- |
Agrocybe pediades |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + O2 | - |
Agrocybe pediades | ? | - |
? | |
| 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + O2 | - |
Agrocybe pediades AH40210 | ? | - |
? | |
| 2,6-dimethoxyphenol + O2 | - |
Agrocybe pediades | ? | - |
? | |
| 2,6-dimethoxyphenol + O2 | - |
Agrocybe pediades AH40210 | ? | - |
? | |
| Evans Blue + O2 | activity with mutant enzyme V159E/N398D | Agrocybe pediades | ? | - |
? | |
| guaiacol + O2 | - |
Agrocybe pediades | ? | - |
? | |
| guaiacol + O2 | - |
Agrocybe pediades AH40210 | ? | - |
? | |
| N,N-dimethyl-1,4-phenylenediamine + O2 | - |
Agrocybe pediades | ? | - |
? | |
| N,N-dimethyl-1,4-phenylenediamine + O2 | - |
Agrocybe pediades AH40210 | ? | - |
? | |
| p-phenylenediamine + O2 | - |
Agrocybe pediades | ? | - |
? | |
| p-phenylenediamine + O2 | - |
Agrocybe pediades AH40210 | ? | - |
? | |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 61.7 | - |
T50-value, mutant enzyme V159E/N398D/I453F/M454L | Agrocybe pediades |
| 62.5 | - |
T50-value, mutant enzyme V159E/N398D/I453L/M454L | Agrocybe pediades |
| 65.8 | - |
T50-value, mutant enzyme V159E | Agrocybe pediades |
| 65.8 | - |
T50-value, mutant enzyme V159E/N398D | Agrocybe pediades |
| 68.8 | - |
T50-value, mutant enzyme N398D | Agrocybe pediades |
| 69.9 | - |
T50-value, wild-type enzyme | Agrocybe pediades |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0046 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | pH 3, temperature not specified in the publication, mutant enzyme V159E/N398D | Agrocybe pediades | |
| 0.08 | - |
2,6-dimethoxyphenol | pH 5, temperature not specified in the publication, mutant enzyme V159E/N398D | Agrocybe pediades | |
| 1.7 | - |
N,N-dimethyl-1,4-phenylenediamine | pH 4, temperature not specified in the publication, mutant enzyme V159E/N398D | Agrocybe pediades | |
| 6 | - |
2,6-dimethoxyphenol | pH 5, temperature not specified in the publication, mutant enzyme V159E | Agrocybe pediades | |
| 33 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | pH 3, temperature not specified in the publication, mutant enzyme V159E | Agrocybe pediades | |
| 56 | - |
N,N-dimethyl-1,4-phenylenediamine | pH 4, temperature not specified in the publication, mutant enzyme V159E | Agrocybe pediades | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 3 | - |
oxidation of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | Agrocybe pediades |
| 4 | - |
oxidation of 2,6-dimethoxyphenol | Agrocybe pediades |
IC50 Value
| IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
|---|---|---|---|---|---|
| 0.051 | - |
pH and temperature not specified in the publication, mutant enzyme V159E/N398D | Agrocybe pediades | NaF | |
| 0.7 | - |
pH and temperature not specified in the publication, mutant enzyme V159E/N398D | Agrocybe pediades | SDS | |
| 74 | - |
pH and temperature not specified in the publication, mutant enzyme V159E/N398D | Agrocybe pediades | NaCl | |
| 90 | - |
pH and temperature not specified in the publication, mutant enzyme V159E/N398D | Agrocybe pediades | EDTA | |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 107 | - |
N,N-dimethyl-1,4-phenylenediamine | pH 4, temperature not specified in the publication, mutant enzyme V159E | Agrocybe pediades | |
| 371 | - |
2,6-dimethoxyphenol | pH 5, temperature not specified in the publication, mutant enzyme V159E | Agrocybe pediades | |
| 388 | - |
N,N-dimethyl-1,4-phenylenediamine | pH 4, temperature not specified in the publication, mutant enzyme V159E/N398D | Agrocybe pediades | |
| 1490 | - |
2,6-dimethoxyphenol | pH 5, temperature not specified in the publication, mutant enzyme V159E/N398D | Agrocybe pediades | |
| 7391 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | pH 3, temperature not specified in the publication, mutant enzyme V159E | Agrocybe pediades | |
| 75500 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | pH 3, temperature not specified in the publication, mutant enzyme V159E/N398D | Agrocybe pediades | |
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