Cloned (Comment) | Organism |
---|---|
gene lhpD, DNA and amino acid sequence determination and analysis, phylogenetic tree, genetic organization of genes encoding enzyme involved in the trans-3-hydroxy-L-proline pathway and networks with D-lysine and D-proline, overview. Recombinant expression of His6-tagged enzyme in Escherichia coli | Pseudomonas aeruginosa |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.447 | - |
1-pyrroline-2-carboxylate | purified recombinant His-tagged enzyme, with NADPH, pH 7.0, 30°C | Pseudomonas aeruginosa | |
0.835 | - |
1-pyrroline-(4R)-hydroxy-2-carboxylate | purified recombinant His-tagged enzyme, with NADPH, pH 7.0, 30°C | Pseudomonas aeruginosa | |
1.57 | - |
1-piperideine-2-carboxylate | purified recombinant His-tagged enzyme, with NADPH, pH 7.0, 30°C | Pseudomonas aeruginosa | |
18.5 | - |
L-proline | purified recombinant His-tagged enzyme, with NADPH, pH 7.0, 30°C | Pseudomonas aeruginosa | |
34.8 | - |
L-pipecolate | purified recombinant His-tagged enzyme, with NADPH, pH 7.0, 30°C | Pseudomonas aeruginosa | |
132 | - |
trans-3-hydroxy-L-proline | purified recombinant His-tagged enzyme, with NADPH, pH 7.0, 30°C | Pseudomonas aeruginosa |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
78000 | - |
recombinant enzyme, gel filtration | Pseudomonas aeruginosa |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
1-piperideine-2-carboxylate + NADPH + H+ | Pseudomonas aeruginosa | - |
L-pipecolate + NADP+ | - |
r | |
1-pyrroline-(4R)-hydroxy-2-carboxylate + NADPH + H+ | Pseudomonas aeruginosa | - |
(4R)-hydroxy-L-proline + NADP+ | - |
r | |
1-pyrroline-2-carboxylate + NADPH + H+ | Pseudomonas aeruginosa | - |
L-proline + NADP+ | - |
r | |
cis-4-hydroxy-L-proline + NADPH + H+ | Pseudomonas aeruginosa | - |
? + NADP+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas aeruginosa | Q9I492 | - |
- |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
additional information | the organism cannot grow on trans-3-hydroxy-L-proline as sole carbon source | Pseudomonas aeruginosa | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.49 | - |
purified recombinant His-tagged enzyme, substrate trans-3-hydroxy-L-proline , pH 7.0, 30°C | Pseudomonas aeruginosa |
0.679 | - |
purified recombinant His-tagged enzyme, substrate L-pipecolate, pH 7.0, 30°C | Pseudomonas aeruginosa |
1.98 | - |
purified recombinant His-tagged enzyme, substrate L-proline, pH 7.0, 30°C | Pseudomonas aeruginosa |
12.1 | - |
purified recombinant His-tagged enzyme, substrates 1-pyrroline-(4R)-hydroxy-2-carboxylate and NADH, pH 7.0, 30°C | Pseudomonas aeruginosa |
42.3 | - |
purified recombinant His-tagged enzyme, substrate 1-piperideine-2-carboxylate, pH 7.0, 30°C | Pseudomonas aeruginosa |
42.3 | - |
purified recombinant His-tagged enzyme, substrate 1-pyrroline-2-carboxylate, pH 7.0, 30°C | Pseudomonas aeruginosa |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1-piperideine-2-carboxylate + NADPH + H+ | - |
Pseudomonas aeruginosa | L-pipecolate + NADP+ | - |
r | |
1-piperideine-2-carboxylate + NADPH + H+ | the reaction equilibrium favors the direction toward NADPH-dependent reduction | Pseudomonas aeruginosa | L-pipecolate + NADP+ | - |
r | |
1-pyrroline-(4R)-hydroxy-2-carboxylate + NADPH + H+ | - |
Pseudomonas aeruginosa | (4R)-hydroxy-L-proline + NADP+ | - |
r | |
1-pyrroline-2-carboxylate + NADPH + H+ | - |
Pseudomonas aeruginosa | L-proline + NADP+ | - |
r | |
1-pyrroline-2-carboxylate + NADPH + H+ | the reaction equilibrium favors the direction toward NADPH-dependent reduction | Pseudomonas aeruginosa | L-proline + NADP+ | - |
r | |
cis-3-hydroxy-L-proline + NADPH + H+ | low activity | Pseudomonas aeruginosa | ? + NADP+ | - |
r | |
cis-4-hydroxy-L-proline + NADPH + H+ | - |
Pseudomonas aeruginosa | ? + NADP+ | - |
r | |
cis-4-hydroxy-L-proline + NADPH + H+ | low activity | Pseudomonas aeruginosa | ? + NADP+ | - |
r | |
L-pipecolate + NADP+ | the reaction equilibrium favors the direction toward NADPH-dependent reduction | Pseudomonas aeruginosa | 1-piperideine-2-carboxylate + NADPH + H+ | - |
r | |
L-proline + NADP+ | the reaction equilibrium favors the direction toward NADPH-dependent reduction | Pseudomonas aeruginosa | 1-pyrroline-2-carboxylate + NADPH + H+ | - |
r | |
additional information | PaLhpD shows 32.8%, 14.4% and 12.2% activity for trans-3-hydroxy-L-proline, cis-4-hydroxy-L-proline, and cis-3-hydroxy-L-proline as a percent of L-proline in the oxidation reaction, respectively. Both L-proline and L-pipecolate undergo NADP+-dependent oxidization, but their kcat/Km values are 500 and 350fold lower than those with 1-pyrroline-2-carboxylate and 1-piperideine-2-carboxylate, respectively. No activity with trans-4-hydroxy-D-proline, D-proline, cis-4-hydroxy-D-proline, and cis-3-hydroxy-D-proline, poor activity with trans-4-hydroxy-L-proline | Pseudomonas aeruginosa | ? | - |
? | |
trans-3-hydroxy-L-proline + NADPH + H+ | moderate activity | Pseudomonas aeruginosa | ? + NADP+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 37165, sequence calculation, 2 * 40000, recombinant enzyme, SDS-PAGE | Pseudomonas aeruginosa |
Synonyms | Comment | Organism |
---|---|---|
DELTA1-pyrroline-2-carboxylate reductase | - |
Pseudomonas aeruginosa |
lhpD | - |
Pseudomonas aeruginosa |
NAD(P)H-dependent DELTA1-pyrroline-2-carboxylate reductase | - |
Pseudomonas aeruginosa |
PaLhpD | - |
Pseudomonas aeruginosa |
Pyr2C reductase | - |
Pseudomonas aeruginosa |
Pyr2C/Pip2C reductase | - |
Pseudomonas aeruginosa |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Pseudomonas aeruginosa |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.25 | - |
L-pipecolate | purified recombinant His-tagged enzyme, with NADPH, pH 7.0, 30°C | Pseudomonas aeruginosa | |
3.42 | - |
L-proline | purified recombinant His-tagged enzyme, with NADPH, pH 7.0, 30°C | Pseudomonas aeruginosa | |
4.53 | - |
trans-3-hydroxy-L-proline | purified recombinant His-tagged enzyme, with NADPH, pH 7.0, 30°C | Pseudomonas aeruginosa | |
14.5 | - |
1-pyrroline-(4R)-hydroxy-2-carboxylate | purified recombinant His-tagged enzyme, with NADPH, pH 7.0, 30°C | Pseudomonas aeruginosa | |
35.3 | - |
1-piperideine-2-carboxylate | purified recombinant His-tagged enzyme, with NADPH, pH 7.0, 30°C | Pseudomonas aeruginosa | |
41.67 | - |
1-pyrroline-2-carboxylate | purified recombinant His-tagged enzyme, with NADPH, pH 7.0, 30°C | Pseudomonas aeruginosa |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
reduction reaction of 1-pyrroline-2-carboxylate and 1-piperideine-2-carboxylate | Pseudomonas aeruginosa |
10 | - |
oxidation reaction of L-proline and L-pipecolate | Pseudomonas aeruginosa |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | no activity with NADH | Pseudomonas aeruginosa | |
NADP+ | - |
Pseudomonas aeruginosa | |
NADPH | - |
Pseudomonas aeruginosa |
General Information | Comment | Organism |
---|---|---|
evolution | enzyme Pyr2C reductase is a member of the ornithine cyclodeaminase/micro-crystallin superfamily and behaves similar to DkpA | Pseudomonas aeruginosa |
metabolism | the enzyme is not involved in the trans-3-hydroxy-L-proline pathway and the trans-4-hydroxy-L-proline pathway | Pseudomonas aeruginosa |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.034 | - |
trans-3-hydroxy-L-proline | purified recombinant His-tagged enzyme, with NADPH, pH 7.0, 30°C | Pseudomonas aeruginosa | |
0.065 | - |
L-pipecolate | purified recombinant His-tagged enzyme, with NADPH, pH 7.0, 30°C | Pseudomonas aeruginosa | |
0.185 | - |
L-proline | purified recombinant His-tagged enzyme, with NADPH, pH 7.0, 30°C | Pseudomonas aeruginosa | |
17.3 | - |
1-pyrroline-(4R)-hydroxy-2-carboxylate | purified recombinant His-tagged enzyme, with NADPH, pH 7.0, 30°C | Pseudomonas aeruginosa | |
22.5 | - |
1-piperideine-2-carboxylate | purified recombinant His-tagged enzyme, with NADPH, pH 7.0, 30°C | Pseudomonas aeruginosa | |
93.2 | - |
1-pyrroline-2-carboxylate | purified recombinant His-tagged enzyme, with NADPH, pH 7.0, 30°C | Pseudomonas aeruginosa |