Protein Variants | Comment | Organism |
---|---|---|
E338A | site-directed mutagenesis, substrate binding of the mutant is unaffected, but kcat is reduced 670fold and catalytic efficiency 220fold compared to the wild-type enzyme. The rate of external aldimine formation in the mutant is similar to that of the wild-type enzyme, but it shows no detectable adenosylcobalamin homolysis upon binding of the physiological substrate | Acetoanaerobium sticklandii |
E338D | site-directed mutagenesis, substrate binding of the mutant is unaffected, but kcat is reduced 380fold and catalytic efficiency 60fold compared to the wild-type enzyme. The rate of external aldimine formation in the mutant is similar to that of the wild-type enzyme, but it shows n detectable adenosylcobalamin homolysis upon binding of the physiological substrate | Acetoanaerobium sticklandii |
E338D | site-directed mutagenesis, substrate binding of the mutant is unaffected, but kcat is reduced 380fold and catalytic efficiency 60fold compared to the wild-type enzyme. The rate of external aldimine formation in the mutant is similar to that of the wild-type enzyme, but it shows no detectable adenosylcobalamin homolysis upon binding of the physiological substrate | Acetoanaerobium sticklandii |
E338Q | site-directed mutagenesis, substrate binding of the mutant is unaffected, but kcat is reduced 90fold and catalytic efficiency 20fold compared to the wild-type enzyme. The rate of external aldimine formation in the mutant is similar to that of the wild-type enzyme, but it shows no detectable adenosylcobalamin homolysis upon binding of the physiological substrate | Acetoanaerobium sticklandii |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | pre-steady-state and stedy-state kinetics of wild-type and mutant enzymes | Acetoanaerobium sticklandii | |
0.03 | - |
D-ornithine | recombinant His-tagged mutant E338A, pH 7.5, 30°C | Acetoanaerobium sticklandii | |
0.043 | - |
D-ornithine | recombinant His-tagged mutant E338D, pH 7.5, 30°C | Acetoanaerobium sticklandii | |
0.061 | - |
D-ornithine | recombinant His-tagged mutant E338Q, pH 7.5, 30°C | Acetoanaerobium sticklandii | |
0.19 | - |
D-ornithine | recombinant His-tagged wild-type enzyme, pH 7.5, 30°C | Acetoanaerobium sticklandii |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-ornithine | Acetoanaerobium sticklandii | - |
(2R,4S)-2,4-diaminopentanoate | - |
? | |
D-ornithine | Acetoanaerobium sticklandii DSM 519 | - |
(2R,4S)-2,4-diaminopentanoate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Acetoanaerobium sticklandii | E3PY95 | beta-subunit | - |
Acetoanaerobium sticklandii | E3PY96 | alpha-subunit | - |
Acetoanaerobium sticklandii DSM 519 | E3PY95 | beta-subunit | - |
Acetoanaerobium sticklandii DSM 519 | E3PY96 | alpha-subunit | - |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
D-ornithine = (2R,4S)-2,4-diaminopentanoate | mechanism, overview, a gradual weakening of the electrostatic energy between the protein and the ribose leads to a progressive increase in the activation energy barrier for adenosylcobalamin Co-C bond homolysis, key role for the conserved polar glutamate residue in controlling the initial generation of radical species | Acetoanaerobium sticklandii | |
D-ornithine = (2R,4S)-2,4-diaminopentanoate | mechanism, overview, a gradual weakening of the electrostatic energy between the protein and the ribose leads to a progressive increase in the activation energy barrier for adenosylcobalamin Co?C bond homolysis, key role for the conserved polar glutamate residue in controlling the initial generation of radical species | Acetoanaerobium sticklandii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-ornithine | - |
Acetoanaerobium sticklandii | (2R,4S)-2,4-diaminopentanoate | - |
? | |
D-ornithine | binding of substrate to the enzyme leads to the formation of an electrostatic interaction between a conserved glutamate side chain and the adenosyl ribose of the adenosylcobalamin cofactor. Residue Glu338 is involved in adenosylcobalamin Co-C bond labilization and catalysis | Acetoanaerobium sticklandii | (2R,4S)-2,4-diaminopentanoate | - |
? | |
D-ornithine | - |
Acetoanaerobium sticklandii DSM 519 | (2R,4S)-2,4-diaminopentanoate | - |
? | |
D-ornithine | binding of substrate to the enzyme leads to the formation of an electrostatic interaction between a conserved glutamate side chain and the adenosyl ribose of the adenosylcobalamin cofactor. Residue Glu338 is involved in adenosylcobalamin Co-C bond labilization and catalysis | Acetoanaerobium sticklandii DSM 519 | (2R,4S)-2,4-diaminopentanoate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
OAM | - |
Acetoanaerobium sticklandii |
ornithine 4,5-aminomutase | - |
Acetoanaerobium sticklandii |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Acetoanaerobium sticklandii |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.032 | - |
D-ornithine | recombinant His-tagged mutant E338D, pH 7.5, 30°C | Acetoanaerobium sticklandii | |
2.9 | - |
D-ornithine | recombinant His-tagged wild-type enzyme, pH 7.5, 30°C | Acetoanaerobium sticklandii | |
4.3 | - |
D-ornithine | recombinant His-tagged mutant E338Q, pH 7.5, 30°C | Acetoanaerobium sticklandii | |
7.6 | - |
D-ornithine | recombinant His-tagged mutant E338A, pH 7.5, 30°C | Acetoanaerobium sticklandii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Acetoanaerobium sticklandii |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
adenosylcobalamin | binding analysis with recombinant wild-type and mutant enzymes, overview | Acetoanaerobium sticklandii |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.07 | - |
D-ornithine | recombinant His-tagged mutant E338Q, pH 7.5, 30°C | Acetoanaerobium sticklandii | |
0.25 | - |
D-ornithine | recombinant His-tagged mutant E338A, pH 7.5, 30°C | Acetoanaerobium sticklandii | |
0.75 | - |
D-ornithine | recombinant His-tagged mutant E338D, pH 7.5, 30°C | Acetoanaerobium sticklandii | |
15.2 | - |
D-ornithine | recombinant His-tagged wild-type enzyme, pH 7.5, 30°C | Acetoanaerobium sticklandii |