Cloned (Comment) | Organism |
---|---|
DNA sequence determination and analysis, overexpression in Escherichia coli, enzyme forms inclusion bodies | Pseudomonas sp. |
Protein Variants | Comment | Organism |
---|---|---|
S207A | site-directed mutagenesis, mutation of Ser207, located within the GXSXG motif of serine esterases, leads to inactivation of the enzyme | Pseudomonas sp. |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
chymotrypsin | partial proteolytic digestion of the recombinant enzyme | Pseudomonas sp. | |
EDTA | removal of bound Ca2+ ions inactivates the enzyme | Pseudomonas sp. |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | at least 12 Ca2+ are very tightly bound to one molecule of holoenzyme, which cannot be removed by dialysis but by EDTA treatment, absolutely required for activity, cannot be substituted by other divalent metal ions | Pseudomonas sp. |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
66000 | - |
1 * 66000, SDS-PAGE | Pseudomonas sp. |
68000 | - |
gel filtration | Pseudomonas sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas sp. | Q9RBY1 | enzyme PML, contains the GXSXG motif | - |
Pseudomonas sp. MIS38 | Q9RBY1 | enzyme PML, contains the GXSXG motif | - |
Purification (Comment) | Organism |
---|---|
recombinant from overexpressing Escherichia coli | Pseudomonas sp. |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
triacylglycerol + H2O = diacylglycerol + a carboxylate | Ser207 is an active site serine | Pseudomonas sp. |
Renatured (Comment) | Organism |
---|---|
recombinant enzyme in inclusion bodies is solubilized by 8 M urea and refolded by removing urea in presence of Ca2+ | Pseudomonas sp. |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
3 | - |
purified recombinant enzyme, substrate 4-nitrophenyl palmitate | Pseudomonas sp. |
6 | - |
purified recombinant enzyme, substrate 4-nitrophenyl acetate | Pseudomonas sp. |
11 | - |
purified recombinant enzyme, substrate 4-nitrophenyl butyrate | Pseudomonas sp. |
25 | - |
purified recombinant enzyme, substrate 4-nitrophenyl myristate | Pseudomonas sp. |
30 | - |
purified recombinant enzyme, substrate 4-nitrophenyl caproate | Pseudomonas sp. |
64 | - |
purified recombinant enzyme, substrate 4-nitrophenyl caprylate | Pseudomonas sp. |
72 | - |
purified recombinant enzyme, substrate 4-nitrophenyl laurate | Pseudomonas sp. |
110 | - |
purified recombinant enzyme, substrate 4-nitrophenyl caprate | Pseudomonas sp. |
400 | - |
purified recombinant enzyme, substrate triacetate | Pseudomonas sp. |
2500 | - |
purified recombinant enzyme, substrate tricaprylate | Pseudomonas sp. |
2600 | - |
purified recombinant enzyme, substrate tricaproate | Pseudomonas sp. |
2800 | - |
purified recombinant enzyme, substrate olive oil | Pseudomonas sp. |
3600 | - |
purified recombinant enzyme, substrate tributyrate | 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. MIS38 | 4-nitrophenol + acetate | - |
? | |
4-nitrophenyl butyrate + H2O | low activity | Pseudomonas sp. | 4-nitrophenol + butyrate | - |
? | |
4-nitrophenyl butyrate + H2O | low activity | Pseudomonas sp. MIS38 | 4-nitrophenol + butyrate | - |
? | |
4-nitrophenyl caprate + H2O | high activity | Pseudomonas sp. | 4-nitrophenol + caprate | - |
? | |
4-nitrophenyl caproate + H2O | high activity | Pseudomonas sp. | 4-nitrophenol + caproate | - |
? | |
4-nitrophenyl caproate + H2O | high activity | Pseudomonas sp. MIS38 | 4-nitrophenol + caproate | - |
? | |
4-nitrophenyl caprylate + H2O | high activity | Pseudomonas sp. | 4-nitrophenol + caprylate | - |
? | |
4-nitrophenyl caprylate + H2O | high activity | Pseudomonas sp. MIS38 | 4-nitrophenol + caprylate | - |
? | |
4-nitrophenyl laurate + H2O | - |
Pseudomonas sp. | 4-nitrophenol + laurate | - |
? | |
4-nitrophenyl myristate + H2O | high activity | Pseudomonas sp. | 4-nitrophenol + myristate | - |
? | |
4-nitrophenyl palmitate + H2O | high activity | Pseudomonas sp. | 4-nitrophenol + palmitate | - |
? | |
glyceryl triacetate + H2O | - |
Pseudomonas sp. | glyceryl diacetate + acetic acid | - |
? | |
glyceryl tributyrate + H2O | - |
Pseudomonas sp. | glyceryl dibutyrate + butyric acid | - |
? | |
glyceryl tricaproate + H2O | - |
Pseudomonas sp. | glyceryl dicaproate + caproic acid | - |
? | |
glyceryl tricaprylate + H2O | - |
Pseudomonas sp. | glyceryl dicaprylate + caprylic acid | - |
? | |
additional information | olive oil is utilized as substrate with high activity | Pseudomonas sp. | ? | - |
? | |
additional information | olive oil is utilized as substrate with high activity | Pseudomonas sp. MIS38 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | 1 * 66000, SDS-PAGE | Pseudomonas sp. |
Synonyms | Comment | Organism |
---|---|---|
More | enzyme belongs to the lipase family I.3 | Pseudomonas sp. |
PML | - |
Pseudomonas sp. |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Pseudomonas sp. |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Pseudomonas sp. |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Pseudomonas sp. | - |
- |
4.4 |