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Literature summary for 3.1.1.3 extracted from

  • Kuwahara, K.; Angkawidjaja, C.; Matsumura, H.; Koga, Y.; Takano, K.; Kanaya, S.
    Importance of the Ca2+-binding sites in the N-catalytic domain of a family I.3 lipase for activity and stability (2008), Protein Eng. Des. Sel., 21, 737-744.
    View publication on PubMed
Search for more literature for 3.1.1.3

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Pseudomonas sp.

Crystallization (Commentary)

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

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

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

Organism UniProt Comment Textmining
Pseudomonas sp.
-
-
-
Pseudomonas sp. MIS38
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Pseudomonas sp.

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

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

Synonyms Comment Organism
PML family I.3 lipase Pseudomonas sp.