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Literature summary for 3.1.1.45 extracted from
- A novel dienelactone hydrolase from the thermoacidophilic archaeon Sulfolobus solfataricus P1: purification, characterization, and expression (2010), Biochim. Biophys. Acta, 1800, 1164-1172.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene dlh, expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Saccharolobus solfataricus |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| C151S | site-directed mutagenesis, almost inactive mutant | Saccharolobus solfataricus |
| D198N | site-directed mutagenesis, almost inactive mutant | Saccharolobus solfataricus |
| H229N | site-directed mutagenesis, almost inactive mutant | Saccharolobus solfataricus |
| S102A | site-directed mutagenesis, the mutant is 3-5fold less active than the wild-type enzyme | Saccharolobus solfataricus |
| S103A | site-directed mutagenesis, 20% reduced activity compared to the wild-type enzyme | Saccharolobus solfataricus |
| S120A | site-directed mutagenesis, the mutant is 3-5fold less active than the wild-type enzyme | Saccharolobus solfataricus |
| S139A | site-directed mutagenesis, the mutant is 3-5fold less active than the wild-type enzyme | Saccharolobus solfataricus |
| S147A | site-directed mutagenesis, 20% reduced activity compared to the wild-type enzyme | Saccharolobus solfataricus |
| S210A | site-directed mutagenesis, the mutant shows similar activity like the wild-type enzyme | Saccharolobus solfataricus |
General Stability
| General Stability | Organism |
|---|---|
| the purified enzyme shows high stability against denaturing agents, including various detergents, urea, and organic solvents | Saccharolobus solfataricus |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 4-chloromercuribenzoate | 78% inhibition at 5 mM | Saccharolobus solfataricus |  |
| diethyl dicarbonate | 71% inhibition at 5 mM | Saccharolobus solfataricus | |
| diisopropylfluorophosphate | complete inhibition at 0.01 mM | Saccharolobus solfataricus | |
| HgCl2 | 92% inhibition at 5 mM | Saccharolobus solfataricus | |
| additional information | no or poor inhibition by eserine, pyridoxal 5'-phosphate, and phenylglyoxal | Saccharolobus solfataricus | |
| paraoxon | complete inhibition at 5 mM | Saccharolobus solfataricus | |
| PMSF | complete inhibition at 0.01 mM | Saccharolobus solfataricus | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | kinetics with other 4-nitrophenyl esters for carboxylesterase activity, overview | Saccharolobus solfataricus | |
| 0.1017 | - |
trans-dienelactone | pH 5.0, 60°C | Saccharolobus solfataricus | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 29000 | - |
SDS-PAGE, gel filtration and MALDI TOF mass spectrometry, recombinant enzyme | Saccharolobus solfataricus |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| dienelactone + H2O | Saccharolobus solfataricus | - |
maleylacetate | - |
? | |
| dienelactone + H2O | Saccharolobus solfataricus P1 | - |
maleylacetate | - |
? | |
| additional information | Saccharolobus solfataricus | the enzyme is a typical dienelactone hydrolase belonging to alpha/beta hydrolase family and containing a catalytic triad composed of Cys151, Asp198, and His229 in the active site | ? | - |
? | |
| additional information | Saccharolobus solfataricus P1 | the enzyme is a typical dienelactone hydrolase belonging to alpha/beta hydrolase family and containing a catalytic triad composed of Cys151, Asp198, and His229 in the active site | ? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Saccharolobus solfataricus | B9WPN8 | gene dlh; DSM1616 | - |
| Saccharolobus solfataricus P1 | B9WPN8 | gene dlh; DSM1616 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant enzyme 150fold from Escherichia coli strain BL21(DE3) by anion exchange and hydrophobic interaction chromatography, dialysis, and another anion exchange chromatography | Saccharolobus solfataricus |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 990 | - |
wild-type enzyme, substrate trans-dienelactone, pH 5.0, 60°C | Saccharolobus solfataricus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| dienelactone + H2O | - |
Saccharolobus solfataricus | maleylacetate | - |
? | |
| dienelactone + H2O | - |
Saccharolobus solfataricus P1 | maleylacetate | - |
? | |
| additional information | the enzyme is a typical dienelactone hydrolase belonging to alpha/beta hydrolase family and containing a catalytic triad composed of Cys151, Asp198, and His229 in the active site | Saccharolobus solfataricus | ? | - |
? | |
| additional information | the enzyme also shows carboxylesterase activity toward 4-nitrophenyl esters ranging from butyrate to laurate | Saccharolobus solfataricus | ? | - |
? | |
| additional information | the enzyme is a typical dienelactone hydrolase belonging to alpha/beta hydrolase family and containing a catalytic triad composed of Cys151, Asp198, and His229 in the active site | Saccharolobus solfataricus P1 | ? | - |
? | |
| additional information | the enzyme also shows carboxylesterase activity toward 4-nitrophenyl esters ranging from butyrate to laurate | Saccharolobus solfataricus P1 | ? | - |
? | |
| trans-dienelactone + H2O | the enzyme displays substrate specificities toward trans-dienelactone, not cis-isomer | Saccharolobus solfataricus | maleylacetate | - |
? | |
| trans-dienelactone + H2O | the enzyme displays substrate specificities toward trans-dienelactone, not cis-isomer | Saccharolobus solfataricus P1 | maleylacetate | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| monomer | 1 * 29000, SDS-PAGE, gel filtration and MALDI TOF mass spectrometry, recombinant enzyme | Saccharolobus solfataricus |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| dienelactone hydrolase | - |
Saccharolobus solfataricus |
| More | the enzyme is a typical dienelactone hydrolase belonging to alpha/beta hydrolase family | Saccharolobus solfataricus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 74 | - |
- |
Saccharolobus solfataricus |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | 90 | assay range | Saccharolobus solfataricus |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 90 | - |
purified enzyme, 50% activity remaining after 50 h | Saccharolobus solfataricus |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 9409 | - |
trans-dienelactone | pH 5.0, 60°C | Saccharolobus solfataricus | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 5 | - |
- |
Saccharolobus solfataricus |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 3 | 9 | assay range | Saccharolobus solfataricus |
General Information
| General Information | Comment | Organism |
|---|---|---|
| physiological function | the hydrolysis of dienelactone plays a key role for the microbial degradation of chloroaromatics via chlorocatechols | Saccharolobus solfataricus |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0925 | - |
trans-dienelactone | pH 5.0, 60°C | Saccharolobus solfataricus | |
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Release 2023.1 (January 2023)
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