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Literature summary for 1.10.3.2 extracted from
- Refolding, characterization, and dye decolorization ability of a highly thermostable laccase from Geobacillus sp. JS12 (2020), Protein Expr. Purif., 173, 105646 .
Application
| Application | Comment | Organism |
|---|---|---|
| industry | the high thermostability makes LacG potentially useful for industrial applications | Geobacillus sp. JS12 |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| lacG is cloned and expressed as a fusion protein with six histidine residues in Escherichia coli BL21 (DE3) cells | Geobacillus sp. JS12 |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 2-mercaptoethanol | 1 mM, 68% inhibition | Geobacillus sp. JS12 |  |
| Acetylacetone | 1 mM, 27% inhibition | Geobacillus sp. JS12 | |
| dithiothreitol | 1 mM, 61% inhibition | Geobacillus sp. JS12 | |
| EDTA | 1 mM, 21% inhibition | Geobacillus sp. JS12 | |
| Fe2+ | 1 mM, reduces LacG activity to 55% | Geobacillus sp. JS12 | |
| L-cysteine | 1 mM, 100% inhibition | Geobacillus sp. JS12 | |
| p-coumaric acid | 1 mM, 25% inhibition | Geobacillus sp. JS12 | |
| SDS | 1 mM, 19% inhibition | Geobacillus sp. JS12 | |
| Sodium azide | 1 mM, 12% inhibition | Geobacillus sp. JS12 | |
| Zn2+ | 1 mM, reduces LacG activity to 48% | Geobacillus sp. JS12 | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 3.53 | - |
2,6-dimethoxyphenol | 70°C, pH 6.5 | Geobacillus sp. JS12 | |
| 3.99 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | 70°C, pH 3.0 | Geobacillus sp. JS12 | |
| 4.23 | - |
guaiacol | 70°C, pH 6.0 | Geobacillus sp. JS12 | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Co2+ | 1 mM, activity is enhanced to 141% | Geobacillus sp. JS12 | |
| copper | multi-copper oxidase | Geobacillus sp. JS12 | |
| Mn2+ | 1 mM, activity is enhanced to 127% | Geobacillus sp. JS12 | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 29000 | - |
gel filtration | Geobacillus sp. JS12 |
| 29800 | - |
calculated from sequence | Geobacillus sp. JS12 |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Geobacillus sp. JS12 | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| the protein is primarily found in inclusion bodies. The resulting insoluble proteins are solubilized with 6 M guanidine HCl and refolded using an on-column refolding procedure | Geobacillus sp. JS12 |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + O2 | - |
Geobacillus sp. JS12 | ? | - |
? | |
| 2,6-dimethoxyphenol + O2 | - |
Geobacillus sp. JS12 | ? | - |
? | |
| guaiacol + O2 | - |
Geobacillus sp. JS12 | ? | - |
? | |
| additional information | LacG decolorizes 43% of Congo red and 14% of Malachite green, and the addition of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) as a redox mediator dramatically increases the dye decolorization efficiency | Geobacillus sp. JS12 | ? | - |
- |
|
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| monomer | 1 * 30000, SDS-PAGE | Geobacillus sp. JS12 |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 80 | - |
substrate: 2,6-dimethoxyphenol | Geobacillus sp. JS12 |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 60 | 95 | the recombinant LacG enzyme shows more than 80% activity within 60-95°C | Geobacillus sp. JS12 |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 90 | - |
2 h, recombinant enzyme retains 85% of its activity | Geobacillus sp. JS12 |
| 95 | - |
half-life: 2 h, recombinant enzyme | Geobacillus sp. JS12 |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.3 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | 70°C, pH 3.0 | Geobacillus sp. JS12 | |
| 38.81 | - |
2,6-dimethoxyphenol | 70°C, pH 6.5 | Geobacillus sp. JS12 | |
| 45.17 | - |
guaiacol | 70°C, pH 6.0 | Geobacillus sp. JS12 | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 3 | - |
substrate: 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | Geobacillus sp. JS12 |
| 6 | - |
substrate: guaiacol | Geobacillus sp. JS12 |
| 6.5 | - |
substrate: 2,6-dimethoxyphenol | Geobacillus sp. JS12 |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 5 | 8.5 | pH 5.0: about 50% of maximal activity, pH 8.5: about 50% of maximal activity, substrate: 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | Geobacillus sp. JS12 |
| 5.5 | 8.5 | pH 5.5: about 75% of maximal activity, pH 8.5: about 50% of maximal activity, substrate: 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | Geobacillus sp. JS12 |
pI Value
| Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|
| Geobacillus sp. JS12 | calculated from sequence | - |
6.07 |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.076 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | 70°C, pH 3.0 | Geobacillus sp. JS12 | |
| 10.68 | - |
guaiacol | 70°C, pH 6.0 | Geobacillus sp. JS12 | |
| 10.99 | - |
2,6-dimethoxyphenol | 70°C, pH 6.5 | Geobacillus sp. JS12 | |
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