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Literature summary for 3.2.1.73 extracted from
- Rational design of disulfide bonds increases thermostability of a mesophilic 1,3-1,4-beta-glucanase from Bacillus terquilensis (2016), PLoS ONE, 11, e0154036 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed in Escherichia coli BL21(DE3) cells | Bacillus tequilensis |
| expression in Escherichia coli | Bacillus tequilensis |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| molecular dynamic simulation and and optimization of residue sites in the calcium region to increase the thermostability | Bacillus tequilensis |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| E46P/S43E/H205P/S40E | 64% increase in specific activity, 20°C increase in optimal enzymatic temperature and a 13.8 °C rise in protein melting temperature compared to wild-type | Bacillus tequilensis |
| N31C/T187C | the mutations significantly enhance the protein thermostability | Bacillus tequilensis |
| N31C/T187C/P102C/N125C | introduction of disulfide pairs N31C-T187C and P102C-N125C, 48.3% increase in half-life value at 60°C and a 4.1°C rise in melting temperature, catalytic properties are similar to wild-type with a decrease in optimal pH | Bacillus tequilensis |
| N31C/T187C/P102C/N125C | the mutant with wild type activity shows a 48.3% increase in half-life value at 60°C and a 4.1°C rise in melting temperature compared to the wild type enzyme | Bacillus tequilensis |
| P102C/N125C | the mutations significantly enhance the protein thermostability | Bacillus tequilensis |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.271 | - |
azo blue barley beta-glucan | mutant E46P/S43E/H205P/S40E, pH 6.5, 40°C | Bacillus tequilensis | |
| 0.2773 | - |
Barley beta-glucan | mutant N31C/T187C, at pH 6.5 and 40°C | Bacillus tequilensis |  |
| 0.2775 | - |
Barley beta-glucan | mutant N31C/T187C/P102C/N125C, at pH 6.5 and 40°C | Bacillus tequilensis | |
| 0.2777 | - |
Barley beta-glucan | mutant P102C/N125C, at pH 6.5 and 40°C | Bacillus tequilensis | |
| 0.2778 | - |
Barley beta-glucan | wild type enzyme, at pH 6.5 and 40°C | Bacillus tequilensis | |
| 0.278 | - |
azo blue barley beta-glucan | mutant N31C/T187C/P102C/N125C, pH 6.5, 40°C | Bacillus tequilensis | |
| 0.278 | - |
azo blue barley beta-glucan | wild-type, pH 6.5, 40°C | Bacillus tequilensis | |
| 0.297 | - |
azo blue barley beta-glucan | wild-type, pH 6.5, 40°C | Bacillus tequilensis |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Bacillus tequilensis | - |
- |
- |
| Bacillus tequilensis CGX 5-1 | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| Ni-NTA column chromatography | Bacillus tequilensis |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 40 | 45 | mutant N31C/T187C/P102C/N125C, pH 6.5, 40°C | Bacillus tequilensis |
| 2490 | - |
wild-type, pH 6.5, 40°C | Bacillus tequilensis |
| 3936 | - |
wild-type, pH 6.5, 40°C | Bacillus tequilensis |
| 4094 | - |
mutant E46P/S43E/H205P/S40E, pH 6.5, 40°C | Bacillus tequilensis |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| azo blue barley beta-glucan + H2O | - |
Bacillus tequilensis | ? | - |
? | |
| azo blue barley beta-glucan + H2O | - |
Bacillus tequilensis CGX 5-1 | ? | - |
? | |
| barley beta-glucan + H2O | - |
Bacillus tequilensis | ? | - |
? | |
| barley beta-glucan + H2O | - |
Bacillus tequilensis CGX 5-1 | ? | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| 1,3-1,4-beta-D-glucan 4-glucanohydrolase | - |
Bacillus tequilensis |
| 1,3-1,4-beta-glucanase | - |
Bacillus tequilensis |
| BglT | - |
Bacillus tequilensis |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 45 | - |
wild-type | Bacillus tequilensis |
| 60 | - |
- |
Bacillus tequilensis |
| 65 | - |
mutant E46P/S43E/H205P/S40E | Bacillus tequilensis |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 50.8 | - |
wild-type, melting temperature | Bacillus tequilensis |
| 54.9 | - |
mutant N31C/T187C/P102C/N125C, melting temperature | Bacillus tequilensis |
| 60 | - |
the melting temperature is 50.8°C. The half-life of the enzyme at 60°C is 59 min. The enzyme retains more than 60% of its activity at up to 75°C and is inactivated at 80°C after 10 min of incubation | Bacillus tequilensis |
| 62 | - |
melting temperature, wild-type | Bacillus tequilensis |
| 70 | - |
1 h, 60% residual activity, mutant E46P/S43E/H205P/S40E | Bacillus tequilensis |
| 76.5 | - |
melting temperature, mutant E46P/S43E/H205P/S40E | Bacillus tequilensis |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 137 | - |
azo blue barley beta-glucan | wild-type, pH 6.5, 40°C | Bacillus tequilensis | |
| 183.6 | - |
Barley beta-glucan | mutant P102C/N125C, at pH 6.5 and 40°C | Bacillus tequilensis | |
| 184.9 | - |
Barley beta-glucan | wild type enzyme, at pH 6.5 and 40°C | Bacillus tequilensis | |
| 185 | - |
azo blue barley beta-glucan | wild-type, pH 6.5, 40°C | Bacillus tequilensis | |
| 186 | - |
azo blue barley beta-glucan | mutant N31C/T187C/P102C/N125C, pH 6.5, 40°C | Bacillus tequilensis | |
| 186.3 | - |
Barley beta-glucan | mutant N31C/T187C/P102C/N125C, at pH 6.5 and 40°C | Bacillus tequilensis | |
| 187 | - |
azo blue barley beta-glucan | mutant E46P/S43E/H205P/S40E, pH 6.5, 40°C | Bacillus tequilensis | |
| 187.6 | - |
Barley beta-glucan | mutant N31C/T187C, at pH 6.5 and 40°C | Bacillus tequilensis | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6 | - |
mutant N31C/T187C/P102C/N125C | Bacillus tequilensis |
| 6.5 | - |
- |
Bacillus tequilensis |
| 6.5 | - |
wild-type | Bacillus tequilensis |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 5.5 | 8 | more than 75% activity between pH 5.5 and 8.0 | Bacillus tequilensis |
pH Stability
| pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|
| 5.5 | 7.5 | the enzyme retains about 60% activity at pH 5.5 and 90% at pH 7.5 after 10 min of incubation | Bacillus tequilensis |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 462 | - |
azo blue barley beta-glucan | wild-type, pH 6.5, 40°C | Bacillus tequilensis | |
| 660.4 | - |
Barley beta-glucan | mutant P102C/N125C, at pH 6.5 and 40°C | Bacillus tequilensis | |
| 665 | - |
azo blue barley beta-glucan | wild-type, pH 6.5, 40°C | Bacillus tequilensis | |
| 665.1 | - |
Barley beta-glucan | wild type enzyme, at pH 6.5 and 40°C | Bacillus tequilensis | |
| 672 | - |
azo blue barley beta-glucan | mutant N31C/T187C/P102C/N125C, pH 6.5, 40°C | Bacillus tequilensis | |
| 672.6 | - |
Barley beta-glucan | mutant N31C/T187C/P102C/N125C, at pH 6.5 and 40°C | Bacillus tequilensis | |
| 677.3 | - |
Barley beta-glucan | mutant N31C/T187C, at pH 6.5 and 40°C | Bacillus tequilensis | |
| 691 | - |
azo blue barley beta-glucan | mutant E46P/S43E/H205P/S40E, pH 6.5, 40°C | Bacillus tequilensis |
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