Cloned (Comment) | Organism |
---|---|
metA, expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
C142A | site-directed mutagenesis, the mutant is inactive | Escherichia coli |
C142S | site-directed mutagenesis, the mutant is almost inactive | Escherichia coli |
E237A | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Escherichia coli |
E237D | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Escherichia coli |
E246A | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Escherichia coli |
E246D | site-directed mutagenesis, the mutant shows reduced catalytic efficiency with L-homoserine, but increased with succinyl-CoA compared to the wild-type enzyme | Escherichia coli |
H235N | site-directed mutagenesis, the mutant is almost inactive | Escherichia coli |
K156L | site-directed mutagenesis, the mutant is inactive | Escherichia coli |
K45A/K46A | site-directed mutagenesis, the mutant is inactive | Escherichia coli |
K45L | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Escherichia coli |
K46L | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Escherichia coli |
R193A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Escherichia coli |
R193A/E246A | site-directed mutagenesis, the mutant shows highly reduced catalytic efficiency with L-homoserine and reduced activity with succinyl-CoA compared to the wild-type enzyme | Escherichia coli |
R193K | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Escherichia coli |
R249A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Escherichia coli |
R249K | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Escherichia coli |
Y238F | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Escherichia coli |
Y238F/E246A | site-directed mutagenesis, the mutant shows highly reduced catalytic efficiency with L-homoserine, but increased with succinyl-CoA compared to the wild-type enzyme | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state kinetics and thermodynamics of wild-type and mutant enzymes, overview | Escherichia coli | |
0.043 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant mutant E246D | Escherichia coli | |
0.044 | - |
L-homoserine | pH 7.5, 25°C, recombinant mutant K45L | Escherichia coli | |
0.05 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant mutant Y238F/E246A | Escherichia coli | |
0.094 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant mutant E246A | Escherichia coli | |
0.15 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant mutant E237A | Escherichia coli | |
0.2 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant mutant R193A/E246A | Escherichia coli | |
0.23 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant mutant R249A | Escherichia coli | |
0.24 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant mutant E237D | Escherichia coli | |
0.28 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant wild-type enzyme | Escherichia coli | |
0.31 | - |
L-homoserine | pH 7.5, 25°C, recombinant mutant R193K | Escherichia coli | |
0.31 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant mutant R249K | Escherichia coli | |
0.35 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant mutant Y238F | Escherichia coli | |
0.36 | - |
L-homoserine | pH 7.5, 25°C, recombinant mutant E237D | Escherichia coli | |
0.38 | - |
L-homoserine | pH 7.5, 25°C, recombinant wild-type enzyme | Escherichia coli | |
0.4 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant mutant R193K | Escherichia coli | |
0.43 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant mutant R193A | Escherichia coli | |
1.1 | - |
L-homoserine | pH 7.5, 25°C, recombinant mutant E237A | Escherichia coli | |
1.1 | - |
L-homoserine | pH 7.5, 25°C, recombinant mutant R193A | Escherichia coli | |
1.15 | - |
L-homoserine | pH 7.5, 25°C, recombinant mutant R249A | Escherichia coli | |
1.51 | - |
L-homoserine | pH 7.5, 25°C, recombinant mutant R249K | Escherichia coli | |
1.78 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant mutant K46L | Escherichia coli | |
2 | - |
L-homoserine | pH 7.5, 25°C, recombinant mutant Y238F | Escherichia coli | |
2.26 | - |
L-homoserine | pH 7.5, 25°C, recombinant mutant K46L | Escherichia coli | |
2.9 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant mutant K45L | Escherichia coli | |
9.1 | - |
L-homoserine | pH 7.5, 25°C, recombinant mutant E246D | Escherichia coli | |
59.1 | - |
L-homoserine | pH 7.5, 25°C, recombinant mutant E246A | Escherichia coli | |
93.8 | - |
L-homoserine | pH 7.5, 25°C, recombinant mutant R193A/E246A | Escherichia coli | |
95.5 | - |
L-homoserine | pH 7.5, 25°C, recombinant mutant Y238F/E246A | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
succinyl-CoA + L-homoserine | Escherichia coli | - |
CoA + O-succinyl-L-homoserine | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by 2 different steps of anion exchange chromatography | Escherichia coli |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
succinyl-CoA + L-homoserine = CoA + O-succinyl-L-homoserine | ping pong catalytic reaction mechanism, roles of essential functional groups, the catalytic triad is formed by the nucleophile Cys142, the essential base His235, and Glu237, required for proper orientation of His235, Lys46 is required for interaction with succinyl-CoA, and Arg193 is involved in binding of L-Homoserine | Escherichia coli |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
recombinant wild-type and mutant enzymes | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
succinyl-CoA + L-homoserine | - |
Escherichia coli | CoA + O-succinyl-L-homoserine | - |
? |
Synonyms | Comment | Organism |
---|---|---|
homoserine succinyltransferase | - |
Escherichia coli |
HST | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.8 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant mutant E237A | Escherichia coli | |
1.2 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant mutant R193A/E246A | Escherichia coli | |
1.26 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant mutant K46L | Escherichia coli | |
2 | 3.7 | succinyl-CoA | pH 7.5, 25°C, recombinant mutant R249K | Escherichia coli | |
3.3 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant mutant R193A | Escherichia coli | |
3.4 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant mutant R193K | Escherichia coli | |
6.7 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant mutant E237D | Escherichia coli | |
6.9 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant mutant E246A | Escherichia coli | |
8.9 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant mutant Y238F/E246A | Escherichia coli | |
10 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant mutant R249A | Escherichia coli | |
13 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant mutant Y238F | Escherichia coli | |
15 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant mutant E246D | Escherichia coli | |
25.7 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant wild-type enzyme | Escherichia coli | |
40.1 | - |
succinyl-CoA | pH 7.5, 25°C, recombinant mutant K45L | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Escherichia coli |