Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli | Pseudomonas sp. |
Crystallization (Comment) | Organism |
---|---|
mutant enzymes D157A and D337A, hanging drop vapour diffusion method, with 10% (w/v) polyethylene glycol 20 K, 0.2 M calcium acetate, 5 mM zinc acetate and 20% (v/v) ethylene glycol | Pseudomonas sp. |
Protein Variants | Comment | Organism |
---|---|---|
D157A | mutant lacks the Ca2+ binding site Ca1, is as stable as the wild type enzyme but exhibits less than 0.4% lipase activity | Pseudomonas sp. |
D275A | mutant lacks the Ca2+ binding site Ca2, is less stable than the wild type protein by approximately 5°C in melting temperature, exhibits weak but significant lipase (25%) activity when compared with the wild type protein | Pseudomonas sp. |
D337A | mutant lacks the Ca2+ binding site Ca3, is less stable than the wild type protein by approximately 5°C in melting temperature but fully active (89% relative lipase activity) | Pseudomonas sp. |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | contains three Ca2+-binding sites in the N-catalytic domain (Ca1-Ca3), the Ca2 site is required to make PML fully active, 10 mM required for activity | Pseudomonas sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas sp. | - |
- |
- |
Pseudomonas sp. MIS38 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
- |
Pseudomonas sp. |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
10 | - |
mutant enzyme D157A, at 30°C, in 25 mM Tris-HCl (pH 7.5) in the presence of 10 mM CaCl2 | Pseudomonas sp. |
710 | - |
mutant enzyme D275A, at 30°C, in 25 mM Tris-HCl (pH 7.5) in the presence of 10 mM CaCl2 | Pseudomonas sp. |
2500 | - |
mutant enzyme D337A, at 30°C, in 25 mM Tris-HCl (pH 7.5) in the presence of 10 mM CaCl2 | Pseudomonas sp. |
2800 | - |
wild type enzyme, at 30°C, in 25 mM Tris-HCl (pH 7.5) in the presence of 10 mM CaCl2 | Pseudomonas sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-nitrophenyl laurate + H2O | - |
Pseudomonas sp. | 4-nitrophenol + laurate | - |
? | |
4-nitrophenyl laurate + H2O | - |
Pseudomonas sp. MIS38 | 4-nitrophenol + laurate | - |
? | |
olive oil + H2O | - |
Pseudomonas sp. | ? | - |
? | |
olive oil + H2O | - |
Pseudomonas sp. MIS38 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
PML | family I.3 lipase | Pseudomonas sp. |