Information on EC 1.5.1.B5 - DELTA1-piperideine-2-carboxylate/DELTA1-pyrroline-2-carboxylate reductase (NADH)

for references in articles please use BRENDA:EC1.5.1.B5
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:


The expected taxonomic range for this enzyme is: Thermococcus litoralis

EC NUMBER
COMMENTARY hide
1.5.1.B5
preliminary BRENDA-supplied EC number
RECOMMENDED NAME
GeneOntology No.
DELTA1-piperideine-2-carboxylate/DELTA1-pyrroline-2-carboxylate reductase (NADH)
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
L-pipecolate + NAD+ = DELTA1-piperideine-2-carboxylate + NADH + H+
show the reaction diagram
L-proline + NAD+ = DELTA1-pyrroline-2-carboxylate + NADH + H+
show the reaction diagram
SYSTEMATIC NAME
IUBMB Comments
L-pipecolate/L-proline:NAD+ 2-oxidoreductase
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
metabolism
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1-pyrroline-2-carboxylate + NADH + H+
L-proline + NAD+
show the reaction diagram
cis-4-hydroxy-L-proline + NAD+
1-pyrroline-4-cis-hydroxy-2-carboxylate + NADPH + H+
show the reaction diagram
cis-4-hydroxy-L-proline + NAD+
?
show the reaction diagram
DELTA1-pyrroline-2-carboxylate + NADH + H+
L-proline + NAD+
show the reaction diagram
L-pipecolate + NAD+
1-piperideine-2-carboxylate + NADH + H+
show the reaction diagram
L-pipecolate + NAD+
DELTA1-piperideine-2-carboxylate + NADH + H+
show the reaction diagram
L-proline + NAD+
DELTA1-pyrroline-2-carboxylate + NADH + H+
show the reaction diagram
H3ZMH3 and H3ZMH4
preference of the reaction equilibrium in the direction toward NADH-dependent reduction
-
-
r
trans-4-hydroxy-L-proline + NAD+
1-pyrroline-4-trans-hydroxy-2-carboxylate + NADPH + H+
show the reaction diagram
-
-
-
-
r
trans-4-hydroxy-L-proline + NAD+
?
show the reaction diagram
H3ZMH3 and H3ZMH4
activity is 70% compared to the activity with L-proline
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
1-pyrroline-2-carboxylate + NADH + H+
L-proline + NAD+
show the reaction diagram
DELTA1-pyrroline-2-carboxylate + NADH + H+
L-proline + NAD+
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.944 - 2.77
1-pyrroline-2-carboxylate
0.944 - 2.77
DELTA1-pyrroline-2-carboxylate
0.608
L-pipecolate
H3ZMH3 and H3ZMH4
pH 6.0, 50C, wild-type enzyme
0.444 - 1.12
L-proline
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
16.58 - 558.3
1-pyrroline-2-carboxylate
16.6 - 558
DELTA1-pyrroline-2-carboxylate
0.2
L-pipecolate
H3ZMH3 and H3ZMH4
pH 6.0, 50C, wild-type enzyme
0.23 - 0.61
L-proline
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
17.57 - 201.6
1-pyrroline-2-carboxylate
17.6 - 201
DELTA1-pyrroline-2-carboxylate
0.33
L-pipecolate
H3ZMH3 and H3ZMH4
pH 6.0, 50C, wild-type enzyme
0.2 - 1.4
L-proline
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
23.9
-
purified recombinant wild-type enzyme, pH 6.5, 50C, substrate 1-pyrroline-2-carboxylate
239
-
purified recombinant mutant V224D/A228K enzyme, pH 6.5, 50C, substrate 1-pyrroline-2-carboxylate
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 6
H3ZMH3 and H3ZMH4
substrates: DELTA1-pyrroline-2-carboxylate + NADH + H+
6.5
-
1-pyrroline-2-carboxylate reduction
11
H3ZMH3 and H3ZMH4
substrates: L-proline + NAD+
11.5
-
L-proline oxidation
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 11
H3ZMH3 and H3ZMH4
pH 5.0: about 40% of maximal activity, pH 11.0: about 80% of maximal activity
10 - 12
H3ZMH3 and H3ZMH4
pH 10.0: about 75% of maximal activity, pH 12.0: about 85% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40000
H3ZMH3 and H3ZMH4
2 * 40000, SDS-PAGE
72000
H3ZMH3 and H3ZMH4
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
-
H3ZMH3 and H3ZMH4
recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3)-RIL by nickel affinity chromatography, ultrafiltration, and gel filtration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
gene lphI, sequence comparisons and phylogenetic analysis, recombinant expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)-RIL
-
His-tagged protein is expressed in Escherichia coli BL21. TlLhpI is a fusion of OCC_00362 and OCC_00367 genes by PCR. The gene from Thermococcus litoralis DSM 5473 consists of two separate open-reading frames (ORFs): OCC_00362 and OCC_00367. When a single A is inserted between A227646 and T227647 in the original genome sequence of T. litoralis DSM 5473, the separate ORFs fuse into a single ORF, and the putative amino acid sequence of is homologous to that of the Thermococcus sibiricus TSIB_0634 gene. Although there is a possibility that this OCD-like protein from Thermococcus litoralis DSM 5473 forms a heterooligomeric structure, the TlLhpI gene is synthesized as a single ORF by sequential steps of PCR using sense and antisense primers and genome DNA of Thermococcus litoralis DSM 5473 as a template
H3ZMH3 and H3ZMH4
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
L52E/V224D/A228K
L52R/V224D/A228K
V224D/A228K
L52E/V224D/A228K
-
DELTA1-pyrroline-2-carboxylate reductase activity is the same as in wild-type enzyme, no alanine dehydrogenase activity; 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
-
L52R/V224D/A228K
-
DELTA1-pyrroline-2-carboxylate reductase activity is the same as in wild-type enzyme, no alanine dehydrogenase activity; 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
-
V224D/A228K
-
12fold enhancement of DELTA1-pyrroline-2-carboxylate reductase activity, 6.7fold enhancement for the oxidization activity for L-proline, no alanine dehydrogenase activity; site-directed mutagenesis, the mutant shows 10fold increased activity for substrate DELTA1-pyrrolidine-2-carboxylate as compared to the wild-type enzyme
-
additional information