BRENDA - Enzyme Database

Engineering of the nonspecific phospholipase C from Bacillus cereus: Replacement of glutamic acid-4 by alanine results in loss of interfacial catalysis and enhanced phosphomonoesterase activity

Tan, C.A.; Roberts, M.F.; Biochemistry 37, 4275-4279 (1998)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
3.1.4.3
transformed into Escherichia coli
Bacillus cereus
Engineering
EC Number
Amino acid exchange
Commentary
Organism
3.1.4.3
E4A
Kunkel method
Bacillus cereus
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
3.1.4.3
phosphatidic acid
monomeric and micellar, poor substrate, inhibitor
Bacillus cereus
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.1.4.3
0.03
-
diheptanoylphosphatidylcholine
micellar substrate, wild-type PLC
Bacillus cereus
3.1.4.3
0.19
-
dihexanoylphosphatidylcholine
monomeric substrate, wild-type PLC
Bacillus cereus
3.1.4.3
1.09
-
diheptanoylphosphatidylcholine
micellar substrate, recombinant PLC
Bacillus cereus
3.1.4.3
1.12
-
dihexanoylphosphatidylcholine
monomeric substrate, recombinant PLC
Bacillus cereus
3.1.4.3
4.2
-
dihexanoylphosphatidic acid
monomeric substrate, recombinant PLC
Bacillus cereus
3.1.4.3
9
-
dihexanoylphosphatidic acid
monomeric substrate, wild-type PLC
Bacillus cereus
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
3.1.4.3
extracellular
-
Bacillus cereus
-
-
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
3.1.4.3
Zn2+
metalloenzyme
Bacillus cereus
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.1.4.3
28500
-
-
Bacillus cereus
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
3.1.4.3
Bacillus cereus
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
3.1.4.3
-
Bacillus cereus
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.1.4.3
diheptanoylphosphatidylcholine + H2O
-
135281
Bacillus cereus
diheptanoylglycerol + phosphorylcholine
-
135281
Bacillus cereus
?
3.1.4.3
dihexanoylphosphatidic acid + H2O
-
135281
Bacillus cereus
?
-
-
-
?
3.1.4.3
dihexanoylphosphatidylcholine + H2O
-
135281
Bacillus cereus
dihexanoylglycerol + phosphorylcholine
-
135281
Bacillus cereus
?
3.1.4.3
phosphatidic acid + H2O
-
135281
Bacillus cereus
?
-
-
-
?
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
3.1.4.3
50
-
stable
Bacillus cereus
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.1.4.3
0.009
-
dihexanoylphosphatidic acid
monomeric substrate, wild-type PLC
Bacillus cereus
3.1.4.3
0.0198
-
dihexanoylphosphatidic acid
monomeric substrate, recombinant PLC
Bacillus cereus
3.1.4.3
4.93
-
dihexanoylphosphatidylcholine
monomeric substrate, recombinant PLC
Bacillus cereus
3.1.4.3
6.78
-
dihexanoylphosphatidylcholine
monomeric substrate, wild-type PLC
Bacillus cereus
3.1.4.3
6.85
-
diheptanoylphosphatidylcholine
micellar substrate, recombinant PLC
Bacillus cereus
3.1.4.3
13.8
-
diheptanoylphosphatidylcholine
micellar substrate, wild-type PLC
Bacillus cereus
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
3.1.4.3
transformed into Escherichia coli
Bacillus cereus
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
3.1.4.3
E4A
Kunkel method
Bacillus cereus
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
3.1.4.3
phosphatidic acid
monomeric and micellar, poor substrate, inhibitor
Bacillus cereus
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.1.4.3
0.03
-
diheptanoylphosphatidylcholine
micellar substrate, wild-type PLC
Bacillus cereus
3.1.4.3
0.19
-
dihexanoylphosphatidylcholine
monomeric substrate, wild-type PLC
Bacillus cereus
3.1.4.3
1.09
-
diheptanoylphosphatidylcholine
micellar substrate, recombinant PLC
Bacillus cereus
3.1.4.3
1.12
-
dihexanoylphosphatidylcholine
monomeric substrate, recombinant PLC
Bacillus cereus
3.1.4.3
4.2
-
dihexanoylphosphatidic acid
monomeric substrate, recombinant PLC
Bacillus cereus
3.1.4.3
9
-
dihexanoylphosphatidic acid
monomeric substrate, wild-type PLC
Bacillus cereus
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
3.1.4.3
extracellular
-
Bacillus cereus
-
-
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
3.1.4.3
Zn2+
metalloenzyme
Bacillus cereus
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.1.4.3
28500
-
-
Bacillus cereus
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
3.1.4.3
-
Bacillus cereus
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.1.4.3
diheptanoylphosphatidylcholine + H2O
-
135281
Bacillus cereus
diheptanoylglycerol + phosphorylcholine
-
135281
Bacillus cereus
?
3.1.4.3
dihexanoylphosphatidic acid + H2O
-
135281
Bacillus cereus
?
-
-
-
?
3.1.4.3
dihexanoylphosphatidylcholine + H2O
-
135281
Bacillus cereus
dihexanoylglycerol + phosphorylcholine
-
135281
Bacillus cereus
?
3.1.4.3
phosphatidic acid + H2O
-
135281
Bacillus cereus
?
-
-
-
?
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
3.1.4.3
50
-
stable
Bacillus cereus
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.1.4.3
0.009
-
dihexanoylphosphatidic acid
monomeric substrate, wild-type PLC
Bacillus cereus
3.1.4.3
0.0198
-
dihexanoylphosphatidic acid
monomeric substrate, recombinant PLC
Bacillus cereus
3.1.4.3
4.93
-
dihexanoylphosphatidylcholine
monomeric substrate, recombinant PLC
Bacillus cereus
3.1.4.3
6.78
-
dihexanoylphosphatidylcholine
monomeric substrate, wild-type PLC
Bacillus cereus
3.1.4.3
6.85
-
diheptanoylphosphatidylcholine
micellar substrate, recombinant PLC
Bacillus cereus
3.1.4.3
13.8
-
diheptanoylphosphatidylcholine
micellar substrate, wild-type PLC
Bacillus cereus