BRENDA - Enzyme Database
show all sequences of 1.5.1.B5

Ornithine cyclodeaminase/micro-crystallin homolog from the hyperthermophilic archaeon Thermococcus litoralis functions as a novel DELTA(1)-pyrroline-2-carboxylate reductase involved in putative trans-3-hydroxy-L-proline metabolism

Watanabe, S.; Tozawa, Y.; Watanabe, Y.; FEBS open bio 4, 617-626 (2014)

Data extracted from this reference:

Cloned(Commentary)
Cloned (Commentary)
Organism
gene lphI, sequence comparisons and phylogenetic analysis, recombinant expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)-RIL
Thermococcus litoralis
Engineering
Protein Variants
Commentary
Organism
L52E/V224D/A228K
site-directed mutagenesis, the mutant shows 2fold increased activity and highly increased specifiicity for substrate DELTA1-pyrrolidine-2-carboxylate as compared to the wild-type enzyme
Thermococcus litoralis
L52R/V224D/A228K
site-directed mutagenesis, the mutant shows slightly increased activity and highly increased specifiicity for substrate DELTA1-pyrrolidine-2-carboxylate and no activity with other substrates in contrast to the wild-type enzyme
Thermococcus litoralis
V224D/A228K
site-directed mutagenesis, the mutant shows 10fold increased activity for substrate DELTA1-pyrrolidine-2-carboxylate as compared to the wild-type enzyme
Thermococcus litoralis
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.944
-
1-pyrroline-2-carboxylate
recombinant wild-type enzyme, pH 6.5, 50°C, with NADH
Thermococcus litoralis
2.77
-
1-pyrroline-2-carboxylate
recombinant mutant V224D/A228K enzyme, pH 6.5, 50°C, with NADH
Thermococcus litoralis
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
1-pyrroline-2-carboxylate + NADH + H+
Thermococcus litoralis
-
L-proline + NAD+
-
-
r
1-pyrroline-2-carboxylate + NADH + H+
Thermococcus litoralis DSM 5473
-
L-proline + NAD+
-
-
r
Organism
Organism
UniProt
Commentary
Textmining
Thermococcus litoralis
-
-
-
Thermococcus litoralis DSM 5473
-
-
-
Purification (Commentary)
Purification (Commentary)
Organism
recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3)-RIL by nickel affinity chromatography, ultrafiltration, and gel filtration
Thermococcus litoralis
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
23.9
-
purified recombinant wild-type enzyme, pH 6.5, 50°C, substrate 1-pyrroline-2-carboxylate
Thermococcus litoralis
239
-
purified recombinant mutant V224D/A228K enzyme, pH 6.5, 50°C, substrate 1-pyrroline-2-carboxylate
Thermococcus litoralis
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
1-pyrroline-2-carboxylate + NADH + H+
-
742532
Thermococcus litoralis
L-proline + NAD+
-
-
-
r
1-pyrroline-2-carboxylate + NADH + H+
highly preferred substrates, preference of the reaction equilibrium in the direction toward NADH-dependent reduction
742532
Thermococcus litoralis
L-proline + NAD+
-
-
-
r
1-pyrroline-2-carboxylate + NADH + H+
-
742532
Thermococcus litoralis DSM 5473
L-proline + NAD+
-
-
-
r
1-pyrroline-2-carboxylate + NADH + H+
highly preferred substrates, preference of the reaction equilibrium in the direction toward NADH-dependent reduction
742532
Thermococcus litoralis DSM 5473
L-proline + NAD+
-
-
-
r
cis-4-hydroxy-L-proline + NAD+
-
742532
Thermococcus litoralis
1-pyrroline-4-cis-hydroxy-2-carboxylate + NADPH + H+
-
-
-
r
cis-4-hydroxy-L-proline + NAD+
-
742532
Thermococcus litoralis DSM 5473
1-pyrroline-4-cis-hydroxy-2-carboxylate + NADPH + H+
-
-
-
r
L-pipecolate + NAD+
-
742532
Thermococcus litoralis
1-piperideine-2-carboxylate + NADH + H+
-
-
-
r
L-pipecolate + NAD+
-
742532
Thermococcus litoralis DSM 5473
1-piperideine-2-carboxylate + NADH + H+
-
-
-
r
additional information
trans-4-hydroxy-L-proline, L-2-aminovalerate, and N-methyl-L-alanine. TlLhpI shows NAD+-dependent oxidation of L-proline. The kcat/Km value in the presence of NADP+ is about 50fold lower than that in the presence of NAD+. In addition to L-proline, several L-proline analogues, including cis-4-hydroxy-L-proline (85%), trans-4-hydroxy-L-proline (70%), and L-pipecolate (49%), are active substrates (relative activity to L-proline in parentheses). The enzyme also shows low activity with pyruvate, oxaloacetate, 2-oxobutyrate, 2-oxovalerate, and 4-methyl-2-oxovalerate in the oxidation reaction direction, overview
742532
Thermococcus litoralis
?
-
-
-
-
additional information
trans-4-hydroxy-L-proline, L-2-aminovalerate, and N-methyl-L-alanine. TlLhpI shows NAD+-dependent oxidation of L-proline. The kcat/Km value in the presence of NADP+ is about 50fold lower than that in the presence of NAD+. In addition to L-proline, several L-proline analogues, including cis-4-hydroxy-L-proline (85%), trans-4-hydroxy-L-proline (70%), and L-pipecolate (49%), are active substrates (relative activity to L-proline in parentheses). The enzyme also shows low activity with pyruvate, oxaloacetate, 2-oxobutyrate, 2-oxovalerate, and 4-methyl-2-oxovalerate in the oxidation reaction direction, overview
742532
Thermococcus litoralis DSM 5473
?
-
-
-
-
trans-4-hydroxy-L-proline + NAD+
-
742532
Thermococcus litoralis
1-pyrroline-4-trans-hydroxy-2-carboxylate + NADPH + H+
-
-
-
r
Synonyms
Synonyms
Commentary
Organism
NADH-dependent Pyr2C reductase
-
Thermococcus litoralis
Pyr2C reductase
-
Thermococcus litoralis
TlLhpI
-
Thermococcus litoralis
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
50
-
assay at
Thermococcus litoralis
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
16.58
-
1-pyrroline-2-carboxylate
recombinant wild-type enzyme, pH 6.5, 50°C, with NADH
Thermococcus litoralis
558.3
-
1-pyrroline-2-carboxylate
recombinant mutant V224D/A228K enzyme, pH 6.5, 50°C, with NADH
Thermococcus litoralis
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6.5
-
1-pyrroline-2-carboxylate reduction
Thermococcus litoralis
11.5
-
L-proline oxidation
Thermococcus litoralis
Cofactor
Cofactor
Commentary
Organism
Structure
NAD+
preferred cofactor
Thermococcus litoralis
NADH
preferred cofactor
Thermococcus litoralis
Cloned(Commentary) (protein specific)
Commentary
Organism
gene lphI, sequence comparisons and phylogenetic analysis, recombinant expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)-RIL
Thermococcus litoralis
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NAD+
preferred cofactor
Thermococcus litoralis
NADH
preferred cofactor
Thermococcus litoralis
Engineering (protein specific)
Protein Variants
Commentary
Organism
L52E/V224D/A228K
site-directed mutagenesis, the mutant shows 2fold increased activity and highly increased specifiicity for substrate DELTA1-pyrrolidine-2-carboxylate as compared to the wild-type enzyme
Thermococcus litoralis
L52R/V224D/A228K
site-directed mutagenesis, the mutant shows slightly increased activity and highly increased specifiicity for substrate DELTA1-pyrrolidine-2-carboxylate and no activity with other substrates in contrast to the wild-type enzyme
Thermococcus litoralis
V224D/A228K
site-directed mutagenesis, the mutant shows 10fold increased activity for substrate DELTA1-pyrrolidine-2-carboxylate as compared to the wild-type enzyme
Thermococcus litoralis
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.944
-
1-pyrroline-2-carboxylate
recombinant wild-type enzyme, pH 6.5, 50°C, with NADH
Thermococcus litoralis
2.77
-
1-pyrroline-2-carboxylate
recombinant mutant V224D/A228K enzyme, pH 6.5, 50°C, with NADH
Thermococcus litoralis
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
1-pyrroline-2-carboxylate + NADH + H+
Thermococcus litoralis
-
L-proline + NAD+
-
-
r
1-pyrroline-2-carboxylate + NADH + H+
Thermococcus litoralis DSM 5473
-
L-proline + NAD+
-
-
r
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3)-RIL by nickel affinity chromatography, ultrafiltration, and gel filtration
Thermococcus litoralis
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
23.9
-
purified recombinant wild-type enzyme, pH 6.5, 50°C, substrate 1-pyrroline-2-carboxylate
Thermococcus litoralis
239
-
purified recombinant mutant V224D/A228K enzyme, pH 6.5, 50°C, substrate 1-pyrroline-2-carboxylate
Thermococcus litoralis
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
1-pyrroline-2-carboxylate + NADH + H+
-
742532
Thermococcus litoralis
L-proline + NAD+
-
-
-
r
1-pyrroline-2-carboxylate + NADH + H+
highly preferred substrates, preference of the reaction equilibrium in the direction toward NADH-dependent reduction
742532
Thermococcus litoralis
L-proline + NAD+
-
-
-
r
1-pyrroline-2-carboxylate + NADH + H+
-
742532
Thermococcus litoralis DSM 5473
L-proline + NAD+
-
-
-
r
1-pyrroline-2-carboxylate + NADH + H+
highly preferred substrates, preference of the reaction equilibrium in the direction toward NADH-dependent reduction
742532
Thermococcus litoralis DSM 5473
L-proline + NAD+
-
-
-
r
cis-4-hydroxy-L-proline + NAD+
-
742532
Thermococcus litoralis
1-pyrroline-4-cis-hydroxy-2-carboxylate + NADPH + H+
-
-
-
r
cis-4-hydroxy-L-proline + NAD+
-
742532
Thermococcus litoralis DSM 5473
1-pyrroline-4-cis-hydroxy-2-carboxylate + NADPH + H+
-
-
-
r
L-pipecolate + NAD+
-
742532
Thermococcus litoralis
1-piperideine-2-carboxylate + NADH + H+
-
-
-
r
L-pipecolate + NAD+
-
742532
Thermococcus litoralis DSM 5473
1-piperideine-2-carboxylate + NADH + H+
-
-
-
r
additional information
trans-4-hydroxy-L-proline, L-2-aminovalerate, and N-methyl-L-alanine. TlLhpI shows NAD+-dependent oxidation of L-proline. The kcat/Km value in the presence of NADP+ is about 50fold lower than that in the presence of NAD+. In addition to L-proline, several L-proline analogues, including cis-4-hydroxy-L-proline (85%), trans-4-hydroxy-L-proline (70%), and L-pipecolate (49%), are active substrates (relative activity to L-proline in parentheses). The enzyme also shows low activity with pyruvate, oxaloacetate, 2-oxobutyrate, 2-oxovalerate, and 4-methyl-2-oxovalerate in the oxidation reaction direction, overview
742532
Thermococcus litoralis
?
-
-
-
-
additional information
trans-4-hydroxy-L-proline, L-2-aminovalerate, and N-methyl-L-alanine. TlLhpI shows NAD+-dependent oxidation of L-proline. The kcat/Km value in the presence of NADP+ is about 50fold lower than that in the presence of NAD+. In addition to L-proline, several L-proline analogues, including cis-4-hydroxy-L-proline (85%), trans-4-hydroxy-L-proline (70%), and L-pipecolate (49%), are active substrates (relative activity to L-proline in parentheses). The enzyme also shows low activity with pyruvate, oxaloacetate, 2-oxobutyrate, 2-oxovalerate, and 4-methyl-2-oxovalerate in the oxidation reaction direction, overview
742532
Thermococcus litoralis DSM 5473
?
-
-
-
-
trans-4-hydroxy-L-proline + NAD+
-
742532
Thermococcus litoralis
1-pyrroline-4-trans-hydroxy-2-carboxylate + NADPH + H+
-
-
-
r
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
50
-
assay at
Thermococcus litoralis
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
16.58
-
1-pyrroline-2-carboxylate
recombinant wild-type enzyme, pH 6.5, 50°C, with NADH
Thermococcus litoralis
558.3
-
1-pyrroline-2-carboxylate
recombinant mutant V224D/A228K enzyme, pH 6.5, 50°C, with NADH
Thermococcus litoralis
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6.5
-
1-pyrroline-2-carboxylate reduction
Thermococcus litoralis
11.5
-
L-proline oxidation
Thermococcus litoralis
General Information
General Information
Commentary
Organism
evolution
the enzyme belongs to the ornithine cyclodeaminase/l-crystallin (OCD/CRYM) superfamily
Thermococcus litoralis
General Information (protein specific)
General Information
Commentary
Organism
evolution
the enzyme belongs to the ornithine cyclodeaminase/l-crystallin (OCD/CRYM) superfamily
Thermococcus litoralis
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
17.57
-
1-pyrroline-2-carboxylate
recombinant wild-type enzyme, pH 6.5, 50°C, with NADH
Thermococcus litoralis
201.6
-
1-pyrroline-2-carboxylate
recombinant mutant V224D/A228K enzyme, pH 6.5, 50°C, with NADH
Thermococcus litoralis
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
17.57
-
1-pyrroline-2-carboxylate
recombinant wild-type enzyme, pH 6.5, 50°C, with NADH
Thermococcus litoralis
201.6
-
1-pyrroline-2-carboxylate
recombinant mutant V224D/A228K enzyme, pH 6.5, 50°C, with NADH
Thermococcus litoralis
Other publictions for EC 1.5.1.B5
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Synonyms
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
729750
Watanabe
Ornithine cyclodeaminase/mu-cr ...
Thermococcus litoralis, Thermococcus litoralis DSM 5473
FEBS Open Bio
4
617-626
2014
-
-
1
-
4
-
-
5
-
-
2
2
-
8
-
-
1
2
-
-
-
-
12
1
3
1
-
-
5
2
2
-
2
-
-
-
-
-
1
2
-
4
-
-
-
-
5
-
-
2
2
-
-
-
1
-
-
-
-
12
1
1
-
-
5
2
2
-
-
-
1
1
-
5
5
742532
Watanabe
Ornithine cyclodeaminase/micr ...
Thermococcus litoralis, Thermococcus litoralis DSM 5473
FEBS open bio
4
617-626
2014
-
-
1
-
3
-
-
2
-
-
-
2
-
10
-
-
1
-
-
-
2
-
11
-
3
1
-
-
2
2
-
-
2
-
-
-
-
-
1
2
-
3
-
-
-
-
2
-
-
-
2
-
-
-
1
-
-
2
-
11
-
1
-
-
2
2
-
-
-
-
1
1
-
2
2