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Literature summary for 1.5.1.21 extracted from

  • Watanabe, S.; Tanimoto, Y.; Yamauchi, S.; Tozawa, Y.; Sawayama, S.; Watanabe, Y.
    Identification and characterization of trans-3-hydroxy-l-proline dehydratase and DELTA1-pyrroline-2-carboxylate reductase involved in trans-3-hydroxy-l-proline metabolism of bacteria (2014), FEBS Open Bio, 4, 240-250 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

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 [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 [Da] Molecular Weight Maximum [Da] Comment Organism
78000
-
recombinant enzyme, gel filtration Pseudomonas aeruginosa

Natural Substrates/ Products (Substrates)

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

Organism UniProt Comment Textmining
Pseudomonas aeruginosa Q9I492
-
-

Source Tissue

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 [micromol/min/mg]

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 and Products (Substrate)

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

Subunits Comment Organism
homodimer 2 * 37165, sequence calculation, 2 * 40000, recombinant enzyme, SDS-PAGE Pseudomonas aeruginosa

Synonyms

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 [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Pseudomonas aeruginosa

Turnover Number [1/s]

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

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

Cofactor Comment Organism Structure
additional information no activity with NADH Pseudomonas aeruginosa
NADP+
-
Pseudomonas aeruginosa
NADPH
-
Pseudomonas aeruginosa

General Information

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 [mM/s]

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