EC Number | Cloned (Comment) | Organism |
---|---|---|
3.1.1.3 | expressed in Escherichia coli BL21 (DE3) cells | Pyrococcus furiosus |
3.1.1.5 | into the vector pGEX-KG and subsequently into pET23a for expression in Escherichia coli BL21DE3 pLysS cells | Pyrococcus furiosus |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
3.1.1.3 | biofuel | the high organic solvent stability and thermal stability suggest that this enzyme may have useful biodiesel-related applications | Pyrococcus furiosus |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
3.1.1.3 | phenylmethylsulfonyl fluoride | remarkable reduction in activity at 1 mM | Pyrococcus furiosus | |
3.1.1.5 | phenylmethylsulfonyl fluoride | PMSF | Pyrococcus furiosus |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
3.1.1.3 | 28000 | - |
recombinant protein, approximately 28000-29000 Da, SDS-PAGE | Pyrococcus furiosus |
3.1.1.3 | 31980 | - |
recombinant protein, MALDI-TOF spectrometry | Pyrococcus furiosus |
3.1.1.3 | 32040 | - |
recombinant protein, calculated from amino acid sequence | Pyrococcus furiosus |
3.1.1.3 | 82000 | - |
dynamic light scattering | Pyrococcus furiosus |
3.1.1.5 | 29000 | - |
determined by SDS-PAGE | Pyrococcus furiosus |
3.1.1.5 | 32040 | - |
calculated | Pyrococcus furiosus |
EC Number | Organic Solvent | Comment | Organism |
---|---|---|---|
3.1.1.3 | acetonitrile | over 50% activity is retained at 70°C in the presence of 25% (v/v) acetonitrile | Pyrococcus furiosus |
3.1.1.5 | acetonitrile | 25%, substrate p-nitrophenyl palmitate, relative activiy 58% | Pyrococcus furiosus |
3.1.1.5 | Ethanol | 25%, substrate p-nitrophenyl palmitate, relative activiy 39% | Pyrococcus furiosus |
3.1.1.5 | Methanol | 25%, substrate p-nitrophenyl palmitate, relative activiy 20% | Pyrococcus furiosus |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
3.1.1.3 | Pyrococcus furiosus | Q8U3I6 | - |
- |
3.1.1.5 | Pyrococcus furiosus | Q8TZI9 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
3.1.1.3 | Ni-NTA affinity column chromatography | Pyrococcus furiosus |
3.1.1.5 | using a Ni-NTA affiniity column | Pyrococcus furiosus |
EC Number | Renatured (Comment) | Organism |
---|---|---|
3.1.1.3 | through heating at 80°C for 30 min in the presence of 6 M Gdm-HCl and 1 M NaCl | Pyrococcus furiosus |
3.1.1.5 | the enzyme is refolded from inclusion bodies | Pyrococcus furiosus |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.1.1.3 | 4-nitrophenyl acetate + H2O | - |
Pyrococcus furiosus | 4-nitrophenol + acetate | - |
? | |
3.1.1.3 | 4-nitrophenyl butyrate + H2O | - |
Pyrococcus furiosus | 4-nitrophenol + butyrate | - |
? | |
3.1.1.3 | 4-nitrophenyl palmitate + H2O | - |
Pyrococcus furiosus | 4-nitrophenol + palmitate | - |
? | |
3.1.1.5 | p-nitrophenyl acetate + H2O | substrate activity assay | Pyrococcus furiosus | p-nitrophenol + acetate | - |
? | |
3.1.1.5 | p-nitrophenyl butyrate | substrate activity assay | Pyrococcus furiosus | p-nitrophenol + butyrate | - |
? | |
3.1.1.5 | p-nitrophenyl palmitate | substrate activity assay | Pyrococcus furiosus | p-nitrophenol + palmitate | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
3.1.1.3 | dimer or trimer | in aqueous solution, dynamic light scattering | Pyrococcus furiosus |
3.1.1.5 | homodimer | prediction, based on the determination of the hydrodynamic radius by dynamic light scattering measurements | Pyrococcus furiosus |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
3.1.1.3 | putative lysophospholipase | the enzyme is confirmed to be an esterase and also shown to be a lipase (hydrolyzing 4-nitrophenyl palmitate), with lipolytic activity being overall about 18 to 20fold lower than esterase activity | Pyrococcus furiosus |
3.1.1.5 | lysophospholipase | - |
Pyrococcus furiosus |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
3.1.1.3 | 70 | - |
using 4-nitrophenyl palmitate as a substrate, for a fixed pH of 8.0 | Pyrococcus furiosus |
3.1.1.5 | 70 | - |
- |
Pyrococcus furiosus |
EC Number | Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|---|
3.1.1.3 | 30 | 80 | - |
Pyrococcus furiosus |
3.1.1.5 | 30 | 80 | conformational changes are negligible over the temperature range of 30-80°C | Pyrococcus furiosus |
EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|---|
3.1.1.3 | 85 | 95 | the lysophospholipase begins to unfold only above 85°C, with unfolding remaining incomplete even at 95°C | Pyrococcus furiosus |
3.1.1.5 | 85 | - |
begins to unfold above 85°C, with unfolding remaining incomplete even at 95°C | Pyrococcus furiosus |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
3.1.1.3 | 7 | - |
using 4-nitrophenyl palmitate as a substrate, for a fixed temperature at 70°C | Pyrococcus furiosus |
3.1.1.5 | 7 | - |
- |
Pyrococcus furiosus |