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Information on EC 1.5.99.B2 - proline dehydrogenase (acceptor) and Organism(s) Thermus thermophilus and UniProt Accession Q72IB8

for references in articles please use BRENDA:EC1.5.99.B2
preliminary BRENDA-supplied EC number
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Thermus thermophilus
UNIPROT: Q72IB8 not found.
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The taxonomic range for the selected organisms is: Thermus thermophilus
The expected taxonomic range for this enzyme is: Archaea, Eukaryota, Bacteria
Synonyms
prodh2, pro dehydrogenase, l-prodh, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proline dehydrogenase
-
PATHWAY SOURCE
PATHWAYS
SYSTEMATIC NAME
IUBMB Comments
L-proline:acceptor oxidoreductase
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
3,4-dehydro-L-proline + acceptor + H2O
? + reduced acceptor
show the reaction diagram
efficient substrate
-
-
?
L-proline + 2,6-dichlorophenolindophenol
(S)-1-pyrroline-5-carboxylate + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
artificial electron acceptor
-
-
?
L-proline + 2,6-dichlorophenolindophenol + H2O
(S)-1-pyrroline-5-carboxylate + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
-
-
-
?
L-proline + acceptor
(S)-1-pyrroline-5-carboxylate + reduced acceptor
show the reaction diagram
-
-
-
?
L-proline + cytochrome c + H2O
(S)-1-pyrroline-5-carboxylate + reduced cytochrome c
show the reaction diagram
-
-
-
?
L-proline + H2O
(S)-1-pyrroline-5-carboxylate + H2O2
show the reaction diagram
-
-
-
?
additional information
?
-
trans-4-hydroxy-L-proline, cis-4-hydroxy-L-proline, L-azetidine-2-carboxylic acid, and L-pipecolinic acid are no substrates of PRODH
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
L-proline + acceptor
(S)-1-pyrroline-5-carboxylate + reduced acceptor
show the reaction diagram
-
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
L-proline
substrate inhibition
L-tetrahydro-2-furoic acid
competitive inhibitor
N-propargylglycine
-
additional information
trans-4-hydroxy-L-proline, cis-4-hydroxy-L-proline, L-azetidine-2-carboxylic acid, and L-pipecolinic acid are no inhibitors of PRODH
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4
3,4-dehydro-L-proline
in 50 mM Tris-HCl, at 25°C
6.5 - 175.1
L-proline
additional information
additional information
Michaelis-Menten and Haldane kinetics
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
75
3,4-dehydro-L-proline
in 50 mM Tris-HCl, at 25°C
0.212 - 42.1
L-proline
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.114 - 2.979
L-proline
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.5
L-lactate
in 50 mM Tris-HCl, at 25°C
2.4
L-Mandelate
in 50 mM Tris-HCl, at 25°C
171 - 825
L-proline
1
L-tetrahydro-2-furoic acid
in 50 mM Tris-HCl, at 25°C
0.2
N-propargylglycine
at 25°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45
assay at, both for recombinant MBP-tagged wild-type and mutant enzyme, kcat increases when the temperature is raised from 25°C to 45°C, the temperature is not increased further due to the instability of the MBP-tag
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25 - 45
remarkable 10fold increase in catalytic efficiency from 25°C to 45°C
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37970
MALDI-TOF mass spectrometry
37980
native enzyme, calculated from sequence of cDNA
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
tetramer
mutant F10E/L12E, mass spectrometry and gel filtration
additional information
MBP-TtProDH and MBP-clipped TtProDH are prone to aggregation through non-native self-association. The hydrophobic N-terminal helix of TtProDH is responsible for the self-association process
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
analysis of the three-dimensional model of TtProDH crystal structure, PDB ID 2G37
sitting drop vapour diffusion method, with 100 mM imidazole (pH 7), 100 mM MgCl2, 17% 2-methyl-2,4-pentanediol, and 5 mM dithiothreitol
sitting drop vapour diffusion method, with 100 mM imidazole buffer at pH 7, 100 mM MgCl2, 14% 2-methyl-2,4-pentanediol, and 5 mM dithiothreitol
from a monodisperse protein solution with detergent n-octyl beta-D-glucopyranoside
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
F10E/L12E
two point mutations in helix alphaA. This helix contains several hydrophobic residues. The recombinant MBP-tagged mutant variant exclusively forms tetramers and exhibits excellent catalytic features over a wide range of temperatures
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
90
PRODH exhibits over 85% residual activity after a 1 h incubation at 90°C and over 60% residual activity after 3 h
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
Ni-NTA column chromatography and Sephadex G-25 gel filtration
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
recombinant overexpression of wild-type and mutant MBP-fusion enzyme in Escherichia coli TOP 10 cells, MBP-TtProDH and MBP-clipped TtProDH are proneto aggregation through non-native self-association
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
White, T.A.; Tanner, J.J.
Cloning, purification and crystallization of Thermus thermophilus proline dehydrogenase
Acta Crystallogr. Sect. F
61
737-739
2005
Thermus thermophilus
Manually annotated by BRENDA team
White, T.A.; Johnson, W.H.; Whitman, C.P.; Tanner, J.J.
Structural basis for the inactivation of Thermus thermophilus proline dehydrogenase by N-propargylglycine
Biochemistry
47
5573-5580
2008
Thermus thermophilus (Q72IB8)
Manually annotated by BRENDA team
White, T.A.; Krishnan, N.; Becker, D.F.; Tanner, J.J.
Structure and kinetics of monofunctional proline dehydrogenase from Thermus thermophilus
J. Biol. Chem.
282
14316-14327
2007
Thermus thermophilus (Q72IB8)
Manually annotated by BRENDA team
Huijbers, M.; van Berkel, W.
A more polar N-terminal helix releases MBP-tagged Thermus thermophilus proline dehydrogenase from tetramer-polymer self-association
J. Mol. Catal. B
134
340-346
2016
Thermus thermophilus (Q72IB8)
-
Manually annotated by BRENDA team