Cloned (Comment) | Organism |
---|---|
gene PsEst1, DNA and amino acid sequence determination and analysis, overexpression of the cold-adapted enzyme in Escherichia coli BL21(DE3), formation of inclusion bodies | Pseudomonas sp. |
General Stability | Organism |
---|---|
Triton X-100 is essential for enzyme stability after solubilization | Pseudomonas sp. |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
69000 | - |
or higher oligomeric state, 4 * 69000, calculation from amino acid sequence | Pseudomonas sp. |
300000 | - |
recombinant enzyme, gel filtration | Pseudomonas sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas sp. | Q8GR22 | cold-adapted enzyme | - |
Pseudomonas sp. B11-1 | Q8GR22 | cold-adapted enzyme | - |
Purification (Comment) | Organism |
---|---|
recombinant enzyme solubilized as denatured protein from inclusion bodies after expression in Escherichia coli, denaturation by 8 M urea, purification to homogeneity | Pseudomonas sp. |
Renatured (Comment) | Organism |
---|---|
renaturation to the catalytically active recombinant enzyme from 8 M urea by dialysis in presence of 0.1% Triton X-100, detergents are essential for the efficient renaturation, 0.05-0.1% Triton X-100 is most efficient followed by 0.1% cholic acid, 30% glycerol and 0.05% CHAPS, and 0.1 M sarcosine, overview | Pseudomonas sp. |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
substrate specificty | Pseudomonas sp. |
48 | - |
purified renatured, recombinant enzyme, substrate p-tolyl octanoate | Pseudomonas sp. |
58.2 | - |
purified renatured, recombinant enzyme, substrate 4-nitrophenyl butyrate | Pseudomonas sp. |
62 | - |
purified renatured, recombinant enzyme, substrate vinyl octanoate | Pseudomonas sp. |
86 | - |
purified renatured, recombinant enzyme, substrate vinyl hexanoate | Pseudomonas sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-nitrophenyl acetate + H2O | - |
Pseudomonas sp. | 4-nitrophenol + acetate | - |
? | |
4-nitrophenyl acetate + H2O | - |
Pseudomonas sp. B11-1 | 4-nitrophenol + acetate | - |
? | |
4-nitrophenyl butyrate + H2O | - |
Pseudomonas sp. | 4-nitrophenol + butyrate | - |
? | |
4-nitrophenyl butyrate + H2O | - |
Pseudomonas sp. B11-1 | 4-nitrophenol + butyrate | - |
? | |
4-nitrophenyl caproate + H2O | - |
Pseudomonas sp. | 4-nitrophenol + caproate | - |
? | |
4-nitrophenyl caprylate + H2O | - |
Pseudomonas sp. | 4-nitrophenol + caprylate | - |
? | |
4-nitrophenyl laurate + H2O | - |
Pseudomonas sp. | 4-nitrophenol + laurate | - |
? | |
4-nitrophenyl palmitate + H2O | - |
Pseudomonas sp. | 4-nitrophenol + palmitate | - |
? | |
4-nitrophenyl valerate + H2O | - |
Pseudomonas sp. | 4-nitrophenol + valerate | - |
? | |
butyl octanoate + H2O | - |
Pseudomonas sp. | butanol + octanoate | - |
? | |
ethyl hexanoate + H2O | - |
Pseudomonas sp. | ethanol + hexanoate | - |
? | |
ethyl laurate + H2O | - |
Pseudomonas sp. | ethanol + laurate | - |
? | |
ethyl myristate + H2O | - |
Pseudomonas sp. | ethanol + myristate | - |
? | |
ethyl octanoate + H2O | - |
Pseudomonas sp. | ethanol + octanoate | - |
? | |
ethyl pelargonate + H2O | - |
Pseudomonas sp. | ethanol + pelargonate | - |
? | |
isopentyl octanoate + H2O | - |
Pseudomonas sp. | isopentanol + octanoate | - |
? | |
isopropyl octanoate + H2O | - |
Pseudomonas sp. | isopropanol + octanoate | - |
? | |
methyl octanoate + H2O | - |
Pseudomonas sp. | methanol + octanoate | - |
? | |
additional information | hydrolysis of vinyl and aryl esters of chain length C4-C6 | Pseudomonas sp. | ? | - |
? | |
additional information | hydrolysis of vinyl and aryl esters of chain length C4-C6 | Pseudomonas sp. B11-1 | ? | - |
? | |
p-tolyl octanoate + H2O | - |
Pseudomonas sp. | p-toluol + octanoate | - |
? | |
pentyl octanoate + H2O | - |
Pseudomonas sp. | pentanol + octanoate | - |
? | |
phenyl acetate + H2O | - |
Pseudomonas sp. | phenol + acetate | - |
? | |
phenyl acetate + H2O | - |
Pseudomonas sp. B11-1 | phenol + acetate | - |
? | |
tributyrin + H2O | - |
Pseudomonas sp. | ? | - |
? | |
tributyrin + H2O | - |
Pseudomonas sp. B11-1 | ? | - |
? | |
vinyl acetate + H2O | - |
Pseudomonas sp. | vinyl alcohol + acetate | - |
? | |
vinyl butyrate + H2O | - |
Pseudomonas sp. | vinyl alcohol + butyrate | - |
? | |
vinyl hexanoate + H2O | - |
Pseudomonas sp. | vinyl alcohol + hexanoate | - |
? | |
vinyl octanoate + H2O | - |
Pseudomonas sp. | vinyl alcohol + octanoate | - |
? | |
vinyl propionate + H2O | - |
Pseudomonas sp. | vinyl alcohol + propionate | - |
? |
Subunits | Comment | Organism |
---|---|---|
tetramer | or higher oligomeric state, 4 * 69000, calculation from amino acid sequence | Pseudomonas sp. |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
additional information | - |
cold-adapted enzyme | Pseudomonas sp. |
45 | - |
about | Pseudomonas sp. |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
20 | 55 | 50% activity at 20°C and 55°C | Pseudomonas sp. |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
60 min, more than 90% remaining activity | Pseudomonas sp. |
50 | - |
30 min, complete loss of activity | Pseudomonas sp. |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
about | Pseudomonas sp. |