EC Number | Cloned (Comment) | Organism |
---|---|---|
1.4.1.18 | expression of His-tagged wild-type and mutant LysDHs in Escherichia coli strain Rosettagami (DE3) | Pyrococcus horikoshii |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
1.4.1.18 | purified recombinant enzyme complexed with NAD+ and a sulfate ion, sitting drop vapor diffusion method, 0.001 ml of 10 mg/ml protein in 100 mM citrate, pH 5.8, 2.1 M ammonium sulfate, and 1 mM NAD+, is mixed with 0.001 ml of reservoir solution, 20°C, X-ray diffraction structure determination and analysis at 2.44 A resolution | Pyrococcus horikoshii |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.4.1.18 | E168A | site-directed mutagenesis | Pyrococcus horikoshii |
1.4.1.18 | E168Q | site-directed mutagenesis | Pyrococcus horikoshii |
1.4.1.18 | R226K | site-directed mutagenesis | Pyrococcus horikoshii |
1.4.1.18 | Y156F | site-directed mutagenesis | Pyrococcus horikoshii |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
1.4.1.18 | 42089 | - |
2 * 42600, recombinant enzyme, SDS-PAGE, 2 * 42089, amino acid sequence | Pyrococcus horikoshii |
1.4.1.18 | 42600 | - |
2 * 42600, recombinant enzyme, SDS-PAGE, 2 * 42089, amino acid sequence | Pyrococcus horikoshii |
1.4.1.18 | 97200 | - |
recombinant enzyme, gel filtration | Pyrococcus horikoshii |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.4.1.18 | (S)-2-aminoadipate 6-semialdehyde | Pyrococcus horikoshii | - |
(S)-2,3,4,5-tetrahydropiperidine-2-carboxylate + H2O | i.e. DELTA1-piperidein-6-carboxylate, spontaneous nonenzymatical reaction | ? | |
1.4.1.18 | (S)-2-aminoadipate 6-semialdehyde | Pyrococcus horikoshii ATCC 700860 | - |
(S)-2,3,4,5-tetrahydropiperidine-2-carboxylate + H2O | i.e. DELTA1-piperidein-6-carboxylate, spontaneous nonenzymatical reaction | ? | |
1.4.1.18 | L-lysine + NAD(P)+ + H2O | Pyrococcus horikoshii | - |
(S)-2-aminoadipate 6-semialdehyde + NAD(P)H + H+ + NH3 | - |
? | |
1.4.1.18 | L-lysine + NAD(P)+ + H2O | Pyrococcus horikoshii ATCC 700860 | - |
(S)-2-aminoadipate 6-semialdehyde + NAD(P)H + H+ + NH3 | - |
? | |
1.4.1.18 | additional information | Pyrococcus horikoshii | the enzyme catalyzes the reductive deamination of the epsilon-amino group and acts as a type of NAD+-dependent amine dehydrogenase producing DELTA1-piperideine-6-carboxylate | ? | - |
? | |
1.4.1.18 | additional information | Pyrococcus horikoshii ATCC 700860 | the enzyme catalyzes the reductive deamination of the epsilon-amino group and acts as a type of NAD+-dependent amine dehydrogenase producing DELTA1-piperideine-6-carboxylate | ? | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.4.1.18 | Pyrococcus horikoshii | O59312 | - |
- |
1.4.1.18 | Pyrococcus horikoshii ATCC 700860 | O59312 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.4.1.18 | refolded recombinant His-tagged wild-type and mutant LysDHs from Escherichia coli strain Rosettagami (DE3) by gel filtration and nickel affinity chromatography | Pyrococcus horikoshii |
EC Number | Reaction | Comment | Organism | Reaction ID |
---|---|---|---|---|
1.4.1.18 | L-lysine + NAD+ + H2O = (S)-2-amino-6-oxohexanoate + NADH + H+ + NH3 | reaction mechanism, overview | Pyrococcus horikoshii |
EC Number | Renatured (Comment) | Organism |
---|---|---|
1.4.1.18 | His-tagged wild-type and mutant LysDHs after recombinant expression in Escherichia coli inclusion bodies are suspended in 20 mM Tris-HCl, pH 7.5, containing 5 mM 2-mercaptoethanol and 4% Triton X-100, and disrupted by sonication, followed by solubilization in 50 mM Tris-HCl, pH 7.5, containing 5 mM 2-mercaptoethanol, 6 M guanidine HCl, 0.2 M NaCl, and 1 mM EDTA. Refolding in buffer containing 0.1 M Tris-HCl, pH 7.5, 5 mM 2-mercaptoethanol, 0.4 M arginine, 2 mM EDTA, and 0.1 mM phenylmethylsulfonyl fluoride and incubation for 36 h at 4°C. Refolded LysDH is heated at 80°C for 10 min and clarified by centrifugation | Pyrococcus horikoshii |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.4.1.18 | (S)-2-aminoadipate 6-semialdehyde | - |
Pyrococcus horikoshii | (S)-2,3,4,5-tetrahydropiperidine-2-carboxylate + H2O | i.e. DELTA1-piperidein-6-carboxylate, spontaneous nonenzymatical reaction | ? | |
1.4.1.18 | (S)-2-aminoadipate 6-semialdehyde | - |
Pyrococcus horikoshii | (S)-2,3,4,5-tetrahydropiperidine-2-carboxylate + H2O | spontaneous reaction | ? | |
1.4.1.18 | (S)-2-aminoadipate 6-semialdehyde | - |
Pyrococcus horikoshii ATCC 700860 | (S)-2,3,4,5-tetrahydropiperidine-2-carboxylate + H2O | i.e. DELTA1-piperidein-6-carboxylate, spontaneous nonenzymatical reaction | ? | |
1.4.1.18 | (S)-2-aminoadipate 6-semialdehyde | - |
Pyrococcus horikoshii ATCC 700860 | (S)-2,3,4,5-tetrahydropiperidine-2-carboxylate + H2O | spontaneous reaction | ? | |
1.4.1.18 | L-lysine + NAD(P)+ + H2O | - |
Pyrococcus horikoshii | (S)-2-aminoadipate 6-semialdehyde + NAD(P)H + H+ + NH3 | - |
? | |
1.4.1.18 | L-lysine + NAD(P)+ + H2O | - |
Pyrococcus horikoshii ATCC 700860 | (S)-2-aminoadipate 6-semialdehyde + NAD(P)H + H+ + NH3 | - |
? | |
1.4.1.18 | additional information | active site structure, overview | Pyrococcus horikoshii | ? | - |
? | |
1.4.1.18 | additional information | the enzyme catalyzes the reductive deamination of the epsilon-amino group and acts as a type of NAD+-dependent amine dehydrogenase producing DELTA1-piperideine-6-carboxylate | Pyrococcus horikoshii | ? | - |
? | |
1.4.1.18 | additional information | active site structure, overview | Pyrococcus horikoshii ATCC 700860 | ? | - |
? | |
1.4.1.18 | additional information | the enzyme catalyzes the reductive deamination of the epsilon-amino group and acts as a type of NAD+-dependent amine dehydrogenase producing DELTA1-piperideine-6-carboxylate | Pyrococcus horikoshii ATCC 700860 | ? | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
1.4.1.18 | dimer | 2 * 42600, recombinant enzyme, SDS-PAGE, 2 * 42089, amino acid sequence | Pyrococcus horikoshii |
1.4.1.18 | More | each monomer consists of a Rossmann fold domain and a C-terminal catalytic domain, and the fold of the catalytic domain showed similarity to that of saccharopine reductase, three-dimensional structure of the enzyme, structure comparisons, overview. Subunit A active site contains a sulfate ion not seen in subunit B. Consequently, subunit A adopts a closed conformation, whereas subunit B adopts an open one. In each subunit, one NAD molecule was bound to the active site in an anti-conformation, indicating that the enzyme makes use of pro-R-specific hydride transfer between the two hydrides at C-4 of NADH with type A specificity | Pyrococcus horikoshii |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.4.1.18 | L-lysine 6-dehydrogenase | - |
Pyrococcus horikoshii |
1.4.1.18 | LysDH | - |
Pyrococcus horikoshii |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.4.1.18 | 95 | - |
about, recombinant enzyme | Pyrococcus horikoshii |
EC Number | Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.4.1.18 | 50 | 95 | maximum activity at about 95°C and an extraordinary drop off in activity with reduction in temperature, the activity at 50°C is only about 1/170th of that at 95°C | Pyrococcus horikoshii |
EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.4.1.18 | 100 | - |
purified recombinant enzyme, half-life is 180 min, the enzyme is highly thermostable and retaining full activity even after incubation for 50 min at 100°C at pH 7.5 | Pyrococcus horikoshii |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
1.4.1.18 | 10 | - |
assay at | Pyrococcus horikoshii |
EC Number | pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|---|
1.4.1.18 | 5 | 12 | the purified recombinant enzyme is stable over a wide range of pH values, losing no activity when incubated at pH values between 5.0 and 12 for 30 min at 50°C, most stable at pH 10.0 | Pyrococcus horikoshii |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.4.1.18 | additional information | both NAD+ and NADP+ are able to serve as the coenzyme for Pyrococcus horikoshii LysDH, although the reaction rate with NADP+ is only 7.8% of that with NAD+, when L-lysine is the substrate | Pyrococcus horikoshii | |
1.4.1.18 | NAD+ | preferred cofactor, binding structure, overview. The enzyme makes use of pro-R-specific hydride transfer between the two hydrides at C-4 of NADH with type A specificity | Pyrococcus horikoshii | |
1.4.1.18 | NADP+ | - |
Pyrococcus horikoshii |