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Literature summary for 3.1.1.3 extracted from

  • Amada, K.; Haruki, M.; Imanaka, T.; Morikawa, M.; Kanaya, S.
    Overproduction in Escherichia coli, purification and characterization of a family I.3 lipase from Pseudomonas sp. MIS38 (2000), Biochim. Biophys. Acta, 1478, 201-210.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
DNA sequence determination and analysis, overexpression in Escherichia coli, enzyme forms inclusion bodies Pseudomonas sp.

Protein Variants

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

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

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 [Da] Molecular Weight Maximum [Da] Comment Organism
66000
-
1 * 66000, SDS-PAGE Pseudomonas sp.
68000
-
gel filtration Pseudomonas sp.

Organism

Organism UniProt Comment Textmining
Pseudomonas sp. Q9RBY1 enzyme PML, contains the GXSXG motif
-
Pseudomonas sp. MIS38 Q9RBY1 enzyme PML, contains the GXSXG motif
-

Purification (Commentary)

Purification (Comment) Organism
recombinant from overexpressing Escherichia coli Pseudomonas sp.

Reaction

Reaction Comment Organism Reaction ID
triacylglycerol + H2O = diacylglycerol + a carboxylate Ser207 is an active site serine Pseudomonas sp.

Renatured (Commentary)

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 [micromol/min/mg]

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 and Products (Substrate)

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

Subunits Comment Organism
monomer 1 * 66000, SDS-PAGE Pseudomonas sp.

Synonyms

Synonyms Comment Organism
More enzyme belongs to the lipase family I.3 Pseudomonas sp.
PML
-
Pseudomonas sp.

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Pseudomonas sp.

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Pseudomonas sp.

pI Value

Organism Comment pI Value Maximum pI Value
Pseudomonas sp.
-
-
4.4