BRENDA - Enzyme Database show

Characterization of biosynthetic enzymes for ectoine as a compatible solute in a moderately halophilic eubacterium, Halomonas elongata

Ono, H.; Sawada, K.; Khunajakr, N.; Tao, T.; Yamamoto, M.; Hiramoto, M.; Shinmyo, A.; Takano, M.; Murooka, Y.; J. Bacteriol. 181, 91-99 (1999)

Data extracted from this reference:

General Stability
EC Number
General Stability
Organism
2.3.1.178
enzyme is stabilized in presence of NaCl. Residual activity after a 3 h incubation at 0°C is 100% in presence of 2 M NaCl
Halomonas elongata
4.2.1.108
NaCl stabilizes the enzyme at above 2 M
Halomonas elongata
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.6.1.46
4.5
-
L-2-amino-4-oxobutanoate
pH 8.5, 25ºC
Halomonas elongata
2.6.1.46
9.1
-
L-glutamate
pH 8.5, 25ºC
Halomonas elongata
2.6.1.76
4.5
-
L-aspartate 4-semialdehyde
pH 8.5, 25°C
Halomonas elongata
2.6.1.76
9.1
-
L-glutamate
pH 8.5, 25°C
Halomonas elongata
4.2.1.108
8.4
-
4-N-acetyl-L-2,4-diaminobutanoate
pH 9.5, 15°C, in presence of 0.77 M NaCl
Halomonas elongata
4.2.1.108
11
-
4-N-acetyl-L-2,4-diaminobutanoate
pH 9.5, 15°C, in presence of 0.05 M NaCl
Halomonas elongata
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
2.6.1.46
K+
required for activity and stability
Halomonas elongata
2.6.1.76
K+
required
Halomonas elongata
4.2.1.108
additional information
the enzyme is not affected by Mg2+, Ca2+, Mn2+, Li+, K+, and NH4+
Halomonas elongata
4.2.1.108
NaCl
required for activity, maximal activity at 0.5 M
Halomonas elongata
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.3.1.178
45000
-
gel filtration
Halomonas elongata
2.6.1.46
44000
-
gel filtration
Halomonas elongata
2.6.1.76
44000
-
6 * 44000, SDS-PAGE
Halomonas elongata
2.6.1.76
260000
-
gel filtration
Halomonas elongata
4.2.1.108
19000
-
x * 19000, SDS-PAGE
Halomonas elongata
4.2.1.108
35000
-
gel filtration in presence of 2 M NaCl
Halomonas elongata
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.6.1.46
L-2-amino-4-oxobutanoate + L-glutamate
Halomonas elongata
-
L-2,4-diaminobutanoate + 2-oxoglutarate
-
Halomonas elongata
r
2.6.1.46
L-2-amino-4-oxobutanoate + L-glutamate
Halomonas elongata OUT300318
-
L-2,4-diaminobutanoate + 2-oxoglutarate
-
Halomonas elongata OUT300318
r
2.6.1.46
L-2-amino-4-oxobutanoate + L-glutamate
Halomonas elongata OUT30018
-
L-2,4-diaminobutanoate + 2-oxoglutarate
-
Halomonas elongata OUT30018
r
4.2.1.108
4-N-acetyl-L-2,4-diaminobutanoate
Halomonas elongata
final step in L-ectoine biosynthesis, pathway overview
L-ectoine + H2O
-
-
?
4.2.1.108
4-N-acetyl-L-2,4-diaminobutanoate
Halomonas elongata OUT300318
final step in L-ectoine biosynthesis, pathway overview
L-ectoine + H2O
-
-
?
4.2.1.108
4-N-acetyl-L-2,4-diaminobutanoate
Halomonas elongata OUT30018
final step in L-ectoine biosynthesis, pathway overview
L-ectoine + H2O
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.3.1.178
Halomonas elongata
-
OUT30018
-
2.3.1.178
Halomonas elongata OUT30018
-
OUT30018
-
2.6.1.46
Halomonas elongata
-
-
-
2.6.1.46
Halomonas elongata OUT300318
-
-
-
2.6.1.76
Halomonas elongata
O52250
-
-
4.2.1.108
Halomonas elongata
-
formerly KS3
-
4.2.1.108
Halomonas elongata OUT30018
-
formerly KS3
-
Purification (Commentary)
EC Number
Commentary
Organism
2.3.1.178
-
Halomonas elongata
2.6.1.46
ammonium sulfate precipitation, Sepharose CL-6B
Halomonas elongata
2.6.1.76
-
Halomonas elongata
4.2.1.108
native enzyme by ammonium sulfate fractionation, hydrophobic interaction chromatography, ultrafiltration, and hydroxylapatite chromatography, in presence of 1 mM L-2,4-diaminobutanoate and over 2 M NaCl to homogeneity
Halomonas elongata
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.3.1.178
50
-
-
Halomonas elongata
2.6.1.46
additional information
-
-
Halomonas elongata
2.6.1.76
12
-
pH 8.5, 25°C
Halomonas elongata
4.2.1.108
16
-
purified enzyme
Halomonas elongata
Storage Stability
EC Number
Storage Stability
Organism
2.6.1.46
0ºC, 100 mM 2,4-diaminobutyric acid
Halomonas elongata
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.3.1.178
acetyl-CoA + L-2,4-diaminobutanoate
-
640091
Halomonas elongata
CoA + N4-acetyl-L-2,4-diaminobutanoate
-
-
-
?
2.3.1.178
acetyl-CoA + L-2,4-diaminobutanoate
-
640091
Halomonas elongata OUT300318
CoA + N4-acetyl-L-2,4-diaminobutanoate
-
-
-
?
2.3.1.178
acetyl-CoA + L-2,4-diaminobutanoate
-
640091
Halomonas elongata OUT30018
CoA + N4-acetyl-L-2,4-diaminobutanoate
-
-
-
?
2.6.1.46
L-2-amino-4-oxobutanoate + L-glutamate
-
640091
Halomonas elongata
L-2,4-diaminobutanoate + 2-oxoglutarate
-
640091
Halomonas elongata
r
2.6.1.46
L-2-amino-4-oxobutanoate + L-glutamate
-
640091
Halomonas elongata OUT300318
L-2,4-diaminobutanoate + 2-oxoglutarate
-
640091
Halomonas elongata OUT300318
r
2.6.1.46
L-2-amino-4-oxobutanoate + L-glutamate
-
640091
Halomonas elongata OUT30018
L-2,4-diaminobutanoate + 2-oxoglutarate
-
640091
Halomonas elongata OUT30018
r
2.6.1.76
4-aminobutanoate + 2-oxoglutarate
about 60% of the activity with L-2,4-diaminobutanoate
640091
Halomonas elongata
? + L-glutamate
-
-
-
r
2.6.1.76
L-2,4-diaminobutanoate + 2-oxoglutarate
-
640091
Halomonas elongata
L-aspartate 4-semialdehyde + L-glutamate
-
-
-
r
2.6.1.76
L-aspartate 4-semialdehyde + L-glutamate
-
640091
Halomonas elongata
L-2,4-diaminobutanoate + 2-oxoglutarate
-
-
-
r
2.6.1.76
L-aspartate 4-semialdehyde + L-homoserine
about 5% of the activity with L-glutamate
640091
Halomonas elongata
L-2,4-diaminobutanoate + 4-hydroxy-2-oxobutanoic acid
-
-
-
r
2.6.1.76
L-ornithine + 2-oxoglutarate
about 6% of the activity with L-2,4-diaminobutanoate
640091
Halomonas elongata
? + L-glutamate
-
-
-
r
2.6.1.76
additional information
no substrates: L-alanine, L-aspartate, L-lysine
640091
Halomonas elongata
?
-
-
-
-
4.2.1.108
4-N-acetyl-L-2,4-diaminobutanoate
final step in L-ectoine biosynthesis, pathway overview
640091
Halomonas elongata
L-ectoine + H2O
-
-
-
?
4.2.1.108
4-N-acetyl-L-2,4-diaminobutanoate
i.e. 1,4,5,6-tetrahydro-2-methyl-4-pyrimidine carboxylic acid, circularization, L-ectoine as an effective osmoprotectant, the enzyme shows absolute substrate specificity, reversibility of the reaction is not detected but might be below detection range
640091
Halomonas elongata
L-ectoine + H2O
-
-
-
?
4.2.1.108
4-N-acetyl-L-2,4-diaminobutanoate
final step in L-ectoine biosynthesis, pathway overview
640091
Halomonas elongata OUT300318
L-ectoine + H2O
-
-
-
?
4.2.1.108
4-N-acetyl-L-2,4-diaminobutanoate
i.e. 1,4,5,6-tetrahydro-2-methyl-4-pyrimidine carboxylic acid, circularization, L-ectoine as an effective osmoprotectant, the enzyme shows absolute substrate specificity, reversibility of the reaction is not detected but might be below detection range
640091
Halomonas elongata OUT300318
L-ectoine + H2O
-
-
-
?
4.2.1.108
4-N-acetyl-L-2,4-diaminobutanoate
final step in L-ectoine biosynthesis, pathway overview
640091
Halomonas elongata OUT30018
L-ectoine + H2O
-
-
-
?
4.2.1.108
4-N-acetyl-L-2,4-diaminobutanoate
i.e. 1,4,5,6-tetrahydro-2-methyl-4-pyrimidine carboxylic acid, circularization, L-ectoine as an effective osmoprotectant, the enzyme shows absolute substrate specificity, reversibility of the reaction is not detected but might be below detection range
640091
Halomonas elongata OUT30018
L-ectoine + H2O
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
2.6.1.76
hexamer
6 * 44000, SDS-PAGE
Halomonas elongata
4.2.1.108
?
x * 19000, SDS-PAGE
Halomonas elongata
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.3.1.178
20
-
-
Halomonas elongata
2.6.1.46
25
-
in presence of 0.4 M NaCl
Halomonas elongata
2.6.1.76
25
-
-
Halomonas elongata
4.2.1.108
-
10
in presence of 0.05 M NaCl
Halomonas elongata
4.2.1.108
15
-
in presence of 0.77 M NaCl
Halomonas elongata
4.2.1.108
30
-
in presence of 3.0 M NaCl
Halomonas elongata
Temperature Range [°C]
EC Number
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
4.2.1.108
-
30
the temperature optimum shifts from 0°C to 30°C with increasing NaCl concentration
Halomonas elongata
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
2.3.1.178
30
-
residual activity after a 3 h incubation at 30°C is 62% in presence of 2 M NaCl
Halomonas elongata
2.3.1.178
37
-
residual activity after a 3 h incubation at 37°C is 35% in presence of 2 M NaCl
Halomonas elongata
4.2.1.108
30
-
the enzyme is quite stable below 30°C in presence of 2 M NaCl
Halomonas elongata
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.3.1.178
8.2
-
-
Halomonas elongata
2.6.1.46
8.6
8.7
-
Halomonas elongata
2.6.1.76
8.6
-
-
Halomonas elongata
4.2.1.108
8.5
9
-
Halomonas elongata
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
2.6.1.76
pyridoxal 5'-phosphate
-
Halomonas elongata
pI Value
EC Number
Organism
Commentary
pI Value Maximum
pI Value
2.6.1.76
Halomonas elongata
isoelectric focussing
-
6.2
4.2.1.108
Halomonas elongata
isoelectric focusing
4.4
4.2
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
2.6.1.76
pyridoxal 5'-phosphate
-
Halomonas elongata
General Stability (protein specific)
EC Number
General Stability
Organism
2.3.1.178
enzyme is stabilized in presence of NaCl. Residual activity after a 3 h incubation at 0°C is 100% in presence of 2 M NaCl
Halomonas elongata
4.2.1.108
NaCl stabilizes the enzyme at above 2 M
Halomonas elongata
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.6.1.46
4.5
-
L-2-amino-4-oxobutanoate
pH 8.5, 25ºC
Halomonas elongata
2.6.1.46
9.1
-
L-glutamate
pH 8.5, 25ºC
Halomonas elongata
2.6.1.76
4.5
-
L-aspartate 4-semialdehyde
pH 8.5, 25°C
Halomonas elongata
2.6.1.76
9.1
-
L-glutamate
pH 8.5, 25°C
Halomonas elongata
4.2.1.108
8.4
-
4-N-acetyl-L-2,4-diaminobutanoate
pH 9.5, 15°C, in presence of 0.77 M NaCl
Halomonas elongata
4.2.1.108
11
-
4-N-acetyl-L-2,4-diaminobutanoate
pH 9.5, 15°C, in presence of 0.05 M NaCl
Halomonas elongata
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
2.6.1.46
K+
required for activity and stability
Halomonas elongata
2.6.1.76
K+
required
Halomonas elongata
4.2.1.108
additional information
the enzyme is not affected by Mg2+, Ca2+, Mn2+, Li+, K+, and NH4+
Halomonas elongata
4.2.1.108
NaCl
required for activity, maximal activity at 0.5 M
Halomonas elongata
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.3.1.178
45000
-
gel filtration
Halomonas elongata
2.6.1.46
44000
-
gel filtration
Halomonas elongata
2.6.1.76
44000
-
6 * 44000, SDS-PAGE
Halomonas elongata
2.6.1.76
260000
-
gel filtration
Halomonas elongata
4.2.1.108
19000
-
x * 19000, SDS-PAGE
Halomonas elongata
4.2.1.108
35000
-
gel filtration in presence of 2 M NaCl
Halomonas elongata
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.6.1.46
L-2-amino-4-oxobutanoate + L-glutamate
Halomonas elongata
-
L-2,4-diaminobutanoate + 2-oxoglutarate
-
Halomonas elongata
r
2.6.1.46
L-2-amino-4-oxobutanoate + L-glutamate
Halomonas elongata OUT300318
-
L-2,4-diaminobutanoate + 2-oxoglutarate
-
Halomonas elongata OUT300318
r
2.6.1.46
L-2-amino-4-oxobutanoate + L-glutamate
Halomonas elongata OUT30018
-
L-2,4-diaminobutanoate + 2-oxoglutarate
-
Halomonas elongata OUT30018
r
4.2.1.108
4-N-acetyl-L-2,4-diaminobutanoate
Halomonas elongata
final step in L-ectoine biosynthesis, pathway overview
L-ectoine + H2O
-
-
?
4.2.1.108
4-N-acetyl-L-2,4-diaminobutanoate
Halomonas elongata OUT300318
final step in L-ectoine biosynthesis, pathway overview
L-ectoine + H2O
-
-
?
4.2.1.108
4-N-acetyl-L-2,4-diaminobutanoate
Halomonas elongata OUT30018
final step in L-ectoine biosynthesis, pathway overview
L-ectoine + H2O
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.3.1.178
-
Halomonas elongata
2.6.1.46
ammonium sulfate precipitation, Sepharose CL-6B
Halomonas elongata
2.6.1.76
-
Halomonas elongata
4.2.1.108
native enzyme by ammonium sulfate fractionation, hydrophobic interaction chromatography, ultrafiltration, and hydroxylapatite chromatography, in presence of 1 mM L-2,4-diaminobutanoate and over 2 M NaCl to homogeneity
Halomonas elongata
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.3.1.178
50
-
-
Halomonas elongata
2.6.1.46
additional information
-
-
Halomonas elongata
2.6.1.76
12
-
pH 8.5, 25°C
Halomonas elongata
4.2.1.108
16
-
purified enzyme
Halomonas elongata
Storage Stability (protein specific)
EC Number
Storage Stability
Organism
2.6.1.46
0ºC, 100 mM 2,4-diaminobutyric acid
Halomonas elongata
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.3.1.178
acetyl-CoA + L-2,4-diaminobutanoate
-
640091
Halomonas elongata
CoA + N4-acetyl-L-2,4-diaminobutanoate
-
-
-
?
2.3.1.178
acetyl-CoA + L-2,4-diaminobutanoate
-
640091
Halomonas elongata OUT300318
CoA + N4-acetyl-L-2,4-diaminobutanoate
-
-
-
?
2.3.1.178
acetyl-CoA + L-2,4-diaminobutanoate
-
640091
Halomonas elongata OUT30018
CoA + N4-acetyl-L-2,4-diaminobutanoate
-
-
-
?
2.6.1.46
L-2-amino-4-oxobutanoate + L-glutamate
-
640091
Halomonas elongata
L-2,4-diaminobutanoate + 2-oxoglutarate
-
640091
Halomonas elongata
r
2.6.1.46
L-2-amino-4-oxobutanoate + L-glutamate
-
640091
Halomonas elongata OUT300318
L-2,4-diaminobutanoate + 2-oxoglutarate
-
640091
Halomonas elongata OUT300318
r
2.6.1.46
L-2-amino-4-oxobutanoate + L-glutamate
-
640091
Halomonas elongata OUT30018
L-2,4-diaminobutanoate + 2-oxoglutarate
-
640091
Halomonas elongata OUT30018
r
2.6.1.76
4-aminobutanoate + 2-oxoglutarate
about 60% of the activity with L-2,4-diaminobutanoate
640091
Halomonas elongata
? + L-glutamate
-
-
-
r
2.6.1.76
L-2,4-diaminobutanoate + 2-oxoglutarate
-
640091
Halomonas elongata
L-aspartate 4-semialdehyde + L-glutamate
-
-
-
r
2.6.1.76
L-aspartate 4-semialdehyde + L-glutamate
-
640091
Halomonas elongata
L-2,4-diaminobutanoate + 2-oxoglutarate
-
-
-
r
2.6.1.76
L-aspartate 4-semialdehyde + L-homoserine
about 5% of the activity with L-glutamate
640091
Halomonas elongata
L-2,4-diaminobutanoate + 4-hydroxy-2-oxobutanoic acid
-
-
-
r
2.6.1.76
L-ornithine + 2-oxoglutarate
about 6% of the activity with L-2,4-diaminobutanoate
640091
Halomonas elongata
? + L-glutamate
-
-
-
r
2.6.1.76
additional information
no substrates: L-alanine, L-aspartate, L-lysine
640091
Halomonas elongata
?
-
-
-
-
4.2.1.108
4-N-acetyl-L-2,4-diaminobutanoate
final step in L-ectoine biosynthesis, pathway overview
640091
Halomonas elongata
L-ectoine + H2O
-
-
-
?
4.2.1.108
4-N-acetyl-L-2,4-diaminobutanoate
i.e. 1,4,5,6-tetrahydro-2-methyl-4-pyrimidine carboxylic acid, circularization, L-ectoine as an effective osmoprotectant, the enzyme shows absolute substrate specificity, reversibility of the reaction is not detected but might be below detection range
640091
Halomonas elongata
L-ectoine + H2O
-
-
-
?
4.2.1.108
4-N-acetyl-L-2,4-diaminobutanoate
final step in L-ectoine biosynthesis, pathway overview
640091
Halomonas elongata OUT300318
L-ectoine + H2O
-
-
-
?
4.2.1.108
4-N-acetyl-L-2,4-diaminobutanoate
i.e. 1,4,5,6-tetrahydro-2-methyl-4-pyrimidine carboxylic acid, circularization, L-ectoine as an effective osmoprotectant, the enzyme shows absolute substrate specificity, reversibility of the reaction is not detected but might be below detection range
640091
Halomonas elongata OUT300318
L-ectoine + H2O
-
-
-
?
4.2.1.108
4-N-acetyl-L-2,4-diaminobutanoate
final step in L-ectoine biosynthesis, pathway overview
640091
Halomonas elongata OUT30018
L-ectoine + H2O
-
-
-
?
4.2.1.108
4-N-acetyl-L-2,4-diaminobutanoate
i.e. 1,4,5,6-tetrahydro-2-methyl-4-pyrimidine carboxylic acid, circularization, L-ectoine as an effective osmoprotectant, the enzyme shows absolute substrate specificity, reversibility of the reaction is not detected but might be below detection range
640091
Halomonas elongata OUT30018
L-ectoine + H2O
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
2.6.1.76
hexamer
6 * 44000, SDS-PAGE
Halomonas elongata
4.2.1.108
?
x * 19000, SDS-PAGE
Halomonas elongata
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.3.1.178
20
-
-
Halomonas elongata
2.6.1.46
25
-
in presence of 0.4 M NaCl
Halomonas elongata
2.6.1.76
25
-
-
Halomonas elongata
4.2.1.108
-
10
in presence of 0.05 M NaCl
Halomonas elongata
4.2.1.108
15
-
in presence of 0.77 M NaCl
Halomonas elongata
4.2.1.108
30
-
in presence of 3.0 M NaCl
Halomonas elongata
Temperature Range [°C] (protein specific)
EC Number
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
4.2.1.108
-
30
the temperature optimum shifts from 0°C to 30°C with increasing NaCl concentration
Halomonas elongata
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
2.3.1.178
30
-
residual activity after a 3 h incubation at 30°C is 62% in presence of 2 M NaCl
Halomonas elongata
2.3.1.178
37
-
residual activity after a 3 h incubation at 37°C is 35% in presence of 2 M NaCl
Halomonas elongata
4.2.1.108
30
-
the enzyme is quite stable below 30°C in presence of 2 M NaCl
Halomonas elongata
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.3.1.178
8.2
-
-
Halomonas elongata
2.6.1.46
8.6
8.7
-
Halomonas elongata
2.6.1.76
8.6
-
-
Halomonas elongata
4.2.1.108
8.5
9
-
Halomonas elongata
pI Value (protein specific)
EC Number
Organism
Commentary
pI Value Maximum
pI Value
2.6.1.76
Halomonas elongata
isoelectric focussing
-
6.2
4.2.1.108
Halomonas elongata
isoelectric focusing
4.4
4.2