BRENDA - Enzyme Database

Purification, characterization, molecular cloning and extracellular production of a phospholipase A1 from Streptomyces albidoflavus NA297

Sugimori, D.; Kano, K.; Matsumoto, Y.; FEBS open bio 2, 318-327 (2012)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
3.1.1.32
gene pla, DNA and amino acid sequence determination and analysis, expression and extracellular production of the recombinant enzyme in Streptomyces lividans from expression vector pUC702/pla
Streptomyces albidoflavus
Engineering
EC Number
Amino acid exchange
Commentary
Organism
3.1.1.32
H218A
site-directed mutagenesis, inactive mutant
Streptomyces albidoflavus
3.1.1.32
H218R
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
Streptomyces albidoflavus
3.1.1.32
S11A
site-directed mutagenesis, inactive mutant
Streptomyces albidoflavus
3.1.1.32
S11D
site-directed mutagenesis, inactive mutant
Streptomyces albidoflavus
3.1.1.32
S11E
site-directed mutagenesis, inactive mutant
Streptomyces albidoflavus
3.1.1.32
S11T
site-directed mutagenesis, inactive mutant
Streptomyces albidoflavus
3.1.1.32
S11Y
site-directed mutagenesis, inactive mutant
Streptomyces albidoflavus
3.1.1.32
S216A
site-directed mutagenesis, inactive mutant
Streptomyces albidoflavus
3.1.1.32
S216D
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
Streptomyces albidoflavus
3.1.1.32
S216E
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
Streptomyces albidoflavus
3.1.1.32
S216T
site-directed mutagenesis, the mutant shows 80-90% reduced activity compared to the wild-type enzyme
Streptomyces albidoflavus
3.1.1.32
S216Y
site-directed mutagenesis, the mutant shows 80-90% reduced activity compared to the wild-type enzyme
Streptomyces albidoflavus
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
3.1.1.32
2-mercaptoethanol
weak inhibition
Streptomyces albidoflavus
3.1.1.32
Ca2+
up to 0.1 mM
Streptomyces albidoflavus
3.1.1.32
Fe2+
-
Streptomyces albidoflavus
3.1.1.32
Fe3+
-
Streptomyces albidoflavus
3.1.1.32
additional information
the enzyme is not inhibited by EDTA and DTT
Streptomyces albidoflavus
3.1.1.32
PMSF
weak inhibition
Streptomyces albidoflavus
3.1.1.32
SDS
-
Streptomyces albidoflavus
3.1.1.32
Triton X-100
weak inhibition at up to 0.23% w/v
Streptomyces albidoflavus
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.1.1.32
2.38
-
phosphatidic acid
pH 7.2, 50C, recombinant enzyme
Streptomyces albidoflavus
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
3.1.1.32
additional information
the enzyme possesses a 33-residue N-terminal signal secretion sequence
Streptomyces albidoflavus
-
-
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
3.1.1.32
Ca2+
activates at 10 mM
Streptomyces albidoflavus
3.1.1.32
additional information
metal ion-independent phospholipase A1
Streptomyces albidoflavus
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.1.1.32
27199
-
1 * 27199, mature protein, seuence calculation, 1 * 28000, recombinant extracellular enzyme, SDS-PAGE
Streptomyces albidoflavus
3.1.1.32
28000
-
DLS analysis
Streptomyces albidoflavus
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
3.1.1.32
Streptomyces albidoflavus
K0J3J2
isolated from Japanese soil, gene pla
-
3.1.1.32
Streptomyces albidoflavus NA297
K0J3J2
isolated from Japanese soil, gene pla
-
Purification (Commentary)
EC Number
Commentary
Organism
3.1.1.32
recombinant extracellular enzyme to homogeneity from the culture supernatant by ammonium sulfate precipitation, hydrophobic interaction chromatography, and anion exchange chromatography
Streptomyces albidoflavus
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
3.1.1.32
588
2873
pH 7.2, 50C, purified recombinant enzyme
Streptomyces albidoflavus
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.1.1.32
1,2-dimyristoyl-sn-glycero-3-phosphate + H2O
-
729743
Streptomyces albidoflavus
?
-
-
-
?
3.1.1.32
1,2-dimyristoyl-sn-glycero-3-phosphate + H2O
-
729743
Streptomyces albidoflavus NA297
?
-
-
-
?
3.1.1.32
1,2-dioleoyl-sn-glycero-3-phosphoethanolamine + H2O
-
729743
Streptomyces albidoflavus
?
-
-
-
?
3.1.1.32
1,2-dioleoyl-sn-glycero-3-phosphoethanolamine + H2O
-
729743
Streptomyces albidoflavus NA297
?
-
-
-
?
3.1.1.32
1,2-dipalmitoyl-sn-glycero-3-phosphocholine + H2O
-
729743
Streptomyces albidoflavus
?
-
-
-
?
3.1.1.32
1,2-dipalmitoyl-sn-glycero-3-phosphocholine + H2O
-
729743
Streptomyces albidoflavus NA297
?
-
-
-
?
3.1.1.32
L-alpha-phosphatidyl-L-serine + H2O
substrate from Glycine max
729743
Streptomyces albidoflavus
2-acyl-1-glycerophosphoserine + a carboxylate
-
-
-
?
3.1.1.32
L-alpha-phosphatidylcholine + H2O
PLA1 activity for egg yolk lecithin hydrolysis, substrate from egg yolk or Glycine max
729743
Streptomyces albidoflavus
2-acylglycerophosphocholine + a carboxylate
-
-
-
?
3.1.1.32
L-alpha-phosphatidylglycerol + H2O
-
729743
Streptomyces albidoflavus
?
-
-
-
?
3.1.1.32
L-alpha-phosphatidylinositol + H2O
-
729743
Streptomyces albidoflavus
2-acyl-sn-glycero-3-phosphoinositol + a carboxylate
-
-
-
?
3.1.1.32
lecithin + H2O
from Glycine max
729743
Streptomyces albidoflavus
?
-
-
-
?
3.1.1.32
additional information
the substrate specificity is pH-dependent, the enzyme preferably hydrolyzes phosphatidic acid and phosphatidylserine at pH 7.2, it is active with lysophosphatidylcholine, but not with triglyceride and the 4-nitrophenyl ester of fatty acids, substrate specificity, overview. At the reaction equilibrium, the molar ratio of released free fatty acids (sn-1:sn-2) is 63:37
729743
Streptomyces albidoflavus
?
-
-
-
-
3.1.1.32
additional information
the substrate specificity is pH-dependent, the enzyme preferably hydrolyzes phosphatidic acid and phosphatidylserine at pH 7.2, it is active with lysophosphatidylcholine, but not with triglyceride and the 4-nitrophenyl ester of fatty acids, substrate specificity, overview. At the reaction equilibrium, the molar ratio of released free fatty acids (sn-1:sn-2) is 63:37
729743
Streptomyces albidoflavus NA297
?
-
-
-
-
3.1.1.32
phosphatidic acid + H2O
-
729743
Streptomyces albidoflavus
2-acyl-lysophosphatidic acid + a carboxylate
-
-
-
?
3.1.1.32
phosphatidic acid + H2O
-
729743
Streptomyces albidoflavus NA297
2-acyl-lysophosphatidic acid + a carboxylate
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
3.1.1.32
monomer
1 * 27199, mature protein, seuence calculation, 1 * 28000, recombinant extracellular enzyme, SDS-PAGE
Streptomyces albidoflavus
Temperature Optimum [C]
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
3.1.1.32
50
-
-
Streptomyces albidoflavus
Temperature Range [C]
EC Number
Temperature Minimum [C]
Temperature Maximum [C]
Commentary
Organism
3.1.1.32
20
70
activity range, profile overview
Streptomyces albidoflavus
Temperature Stability [C]
EC Number
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
3.1.1.32
40
-
native and recombinant enzymes, stable at
Streptomyces albidoflavus
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.1.1.32
630
-
phosphatidic acid
pH 7.2, 50C, recombinant enzyme
Streptomyces albidoflavus
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.1.1.32
7.2
-
-
Streptomyces albidoflavus
pH Range
EC Number
pH Minimum
pH Maximum
Commentary
Organism
3.1.1.32
4
11
activity range, profile overview
Streptomyces albidoflavus
pH Stability
EC Number
pH Stability
pH Stability Maximum
Commentary
Organism
3.1.1.32
5.6
9
recombinant enzyme, stable at
Streptomyces albidoflavus
3.1.1.32
7.2
9
native enzyme, stable at
Streptomyces albidoflavus
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
3.1.1.32
gene pla, DNA and amino acid sequence determination and analysis, expression and extracellular production of the recombinant enzyme in Streptomyces lividans from expression vector pUC702/pla
Streptomyces albidoflavus
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
3.1.1.32
H218A
site-directed mutagenesis, inactive mutant
Streptomyces albidoflavus
3.1.1.32
H218R
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
Streptomyces albidoflavus
3.1.1.32
S11A
site-directed mutagenesis, inactive mutant
Streptomyces albidoflavus
3.1.1.32
S11D
site-directed mutagenesis, inactive mutant
Streptomyces albidoflavus
3.1.1.32
S11E
site-directed mutagenesis, inactive mutant
Streptomyces albidoflavus
3.1.1.32
S11T
site-directed mutagenesis, inactive mutant
Streptomyces albidoflavus
3.1.1.32
S11Y
site-directed mutagenesis, inactive mutant
Streptomyces albidoflavus
3.1.1.32
S216A
site-directed mutagenesis, inactive mutant
Streptomyces albidoflavus
3.1.1.32
S216D
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
Streptomyces albidoflavus
3.1.1.32
S216E
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
Streptomyces albidoflavus
3.1.1.32
S216T
site-directed mutagenesis, the mutant shows 80-90% reduced activity compared to the wild-type enzyme
Streptomyces albidoflavus
3.1.1.32
S216Y
site-directed mutagenesis, the mutant shows 80-90% reduced activity compared to the wild-type enzyme
Streptomyces albidoflavus
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
3.1.1.32
2-mercaptoethanol
weak inhibition
Streptomyces albidoflavus
3.1.1.32
Ca2+
up to 0.1 mM
Streptomyces albidoflavus
3.1.1.32
Fe2+
-
Streptomyces albidoflavus
3.1.1.32
Fe3+
-
Streptomyces albidoflavus
3.1.1.32
additional information
the enzyme is not inhibited by EDTA and DTT
Streptomyces albidoflavus
3.1.1.32
PMSF
weak inhibition
Streptomyces albidoflavus
3.1.1.32
SDS
-
Streptomyces albidoflavus
3.1.1.32
Triton X-100
weak inhibition at up to 0.23% w/v
Streptomyces albidoflavus
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.1.1.32
2.38
-
phosphatidic acid
pH 7.2, 50C, recombinant enzyme
Streptomyces albidoflavus
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
3.1.1.32
additional information
the enzyme possesses a 33-residue N-terminal signal secretion sequence
Streptomyces albidoflavus
-
-
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
3.1.1.32
Ca2+
activates at 10 mM
Streptomyces albidoflavus
3.1.1.32
additional information
metal ion-independent phospholipase A1
Streptomyces albidoflavus
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.1.1.32
27199
-
1 * 27199, mature protein, seuence calculation, 1 * 28000, recombinant extracellular enzyme, SDS-PAGE
Streptomyces albidoflavus
3.1.1.32
28000
-
DLS analysis
Streptomyces albidoflavus
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
3.1.1.32
recombinant extracellular enzyme to homogeneity from the culture supernatant by ammonium sulfate precipitation, hydrophobic interaction chromatography, and anion exchange chromatography
Streptomyces albidoflavus
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
3.1.1.32
588
2873
pH 7.2, 50C, purified recombinant enzyme
Streptomyces albidoflavus
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.1.1.32
1,2-dimyristoyl-sn-glycero-3-phosphate + H2O
-
729743
Streptomyces albidoflavus
?
-
-
-
?
3.1.1.32
1,2-dimyristoyl-sn-glycero-3-phosphate + H2O
-
729743
Streptomyces albidoflavus NA297
?
-
-
-
?
3.1.1.32
1,2-dioleoyl-sn-glycero-3-phosphoethanolamine + H2O
-
729743
Streptomyces albidoflavus
?
-
-
-
?
3.1.1.32
1,2-dioleoyl-sn-glycero-3-phosphoethanolamine + H2O
-
729743
Streptomyces albidoflavus NA297
?
-
-
-
?
3.1.1.32
1,2-dipalmitoyl-sn-glycero-3-phosphocholine + H2O
-
729743
Streptomyces albidoflavus
?
-
-
-
?
3.1.1.32
1,2-dipalmitoyl-sn-glycero-3-phosphocholine + H2O
-
729743
Streptomyces albidoflavus NA297
?
-
-
-
?
3.1.1.32
L-alpha-phosphatidyl-L-serine + H2O
substrate from Glycine max
729743
Streptomyces albidoflavus
2-acyl-1-glycerophosphoserine + a carboxylate
-
-
-
?
3.1.1.32
L-alpha-phosphatidylcholine + H2O
PLA1 activity for egg yolk lecithin hydrolysis, substrate from egg yolk or Glycine max
729743
Streptomyces albidoflavus
2-acylglycerophosphocholine + a carboxylate
-
-
-
?
3.1.1.32
L-alpha-phosphatidylglycerol + H2O
-
729743
Streptomyces albidoflavus
?
-
-
-
?
3.1.1.32
L-alpha-phosphatidylinositol + H2O
-
729743
Streptomyces albidoflavus
2-acyl-sn-glycero-3-phosphoinositol + a carboxylate
-
-
-
?
3.1.1.32
lecithin + H2O
from Glycine max
729743
Streptomyces albidoflavus
?
-
-
-
?
3.1.1.32
additional information
the substrate specificity is pH-dependent, the enzyme preferably hydrolyzes phosphatidic acid and phosphatidylserine at pH 7.2, it is active with lysophosphatidylcholine, but not with triglyceride and the 4-nitrophenyl ester of fatty acids, substrate specificity, overview. At the reaction equilibrium, the molar ratio of released free fatty acids (sn-1:sn-2) is 63:37
729743
Streptomyces albidoflavus
?
-
-
-
-
3.1.1.32
additional information
the substrate specificity is pH-dependent, the enzyme preferably hydrolyzes phosphatidic acid and phosphatidylserine at pH 7.2, it is active with lysophosphatidylcholine, but not with triglyceride and the 4-nitrophenyl ester of fatty acids, substrate specificity, overview. At the reaction equilibrium, the molar ratio of released free fatty acids (sn-1:sn-2) is 63:37
729743
Streptomyces albidoflavus NA297
?
-
-
-
-
3.1.1.32
phosphatidic acid + H2O
-
729743
Streptomyces albidoflavus
2-acyl-lysophosphatidic acid + a carboxylate
-
-
-
?
3.1.1.32
phosphatidic acid + H2O
-
729743
Streptomyces albidoflavus NA297
2-acyl-lysophosphatidic acid + a carboxylate
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
3.1.1.32
monomer
1 * 27199, mature protein, seuence calculation, 1 * 28000, recombinant extracellular enzyme, SDS-PAGE
Streptomyces albidoflavus
Temperature Optimum [C] (protein specific)
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
3.1.1.32
50
-
-
Streptomyces albidoflavus
Temperature Range [C] (protein specific)
EC Number
Temperature Minimum [C]
Temperature Maximum [C]
Commentary
Organism
3.1.1.32
20
70
activity range, profile overview
Streptomyces albidoflavus
Temperature Stability [C] (protein specific)
EC Number
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
3.1.1.32
40
-
native and recombinant enzymes, stable at
Streptomyces albidoflavus
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.1.1.32
630
-
phosphatidic acid
pH 7.2, 50C, recombinant enzyme
Streptomyces albidoflavus
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.1.1.32
7.2
-
-
Streptomyces albidoflavus
pH Range (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
Organism
3.1.1.32
4
11
activity range, profile overview
Streptomyces albidoflavus
pH Stability (protein specific)
EC Number
pH Stability
pH Stability Maximum
Commentary
Organism
3.1.1.32
5.6
9
recombinant enzyme, stable at
Streptomyces albidoflavus
3.1.1.32
7.2
9
native enzyme, stable at
Streptomyces albidoflavus
General Information
EC Number
General Information
Commentary
Organism
3.1.1.32
additional information
residue Ser11 is essential for the catalytic function of the enzyme, the active site may include residues Ser216 and His218
Streptomyces albidoflavus
General Information (protein specific)
EC Number
General Information
Commentary
Organism
3.1.1.32
additional information
residue Ser11 is essential for the catalytic function of the enzyme, the active site may include residues Ser216 and His218
Streptomyces albidoflavus
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
3.1.1.32
265
-
phosphatidic acid
pH 7.2, 50C, recombinant enzyme
Streptomyces albidoflavus
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
3.1.1.32
265
-
phosphatidic acid
pH 7.2, 50C, recombinant enzyme
Streptomyces albidoflavus