Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Monaghan, P.J.; Leys, D.; Scrutton, N.S.
    Mechanistic aspects and redox properties of hyperthermophilic L-proline dehydrogenase from Pyrococcus furiosus related to dimethylglycine dehydrogenase/oxidase (2007), FEBS J., 274, 2070-2087.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.5.99.B2 expressed in Escherichia coli Rosetta(DE3)pLysS cells Pyrococcus furiosus

Protein Variants

EC Number Protein Variants Comment Organism
1.5.99.B2 H225A the mutant is unstable and precipitates from solution below pH 7.0, decreased activity compared to the wild type enzyme Pyrococcus furiosus
1.5.99.B2 H225Q the mutant displays activity down to solution pH 6.0, increased activity compared to the wild type enzyme Pyrococcus furiosus
1.5.99.B2 Y251F wild type pH stability, increased activity compared to the wild type enzyme Pyrococcus furiosus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.5.99.B2 1.95
-
 L-proline mutant enzyme Y215F, in 100 mM potassium phosphate buffer, pH 7.5, at 60°C Pyrococcus furiosus
1.5.99.B2 5.3
-
 L-proline wild type enzyme, in 100 mM potassium phosphate buffer, pH 7.5, at 50°C Pyrococcus furiosus
1.5.99.B2 5.6
-
 L-proline wild type enzyme, in 100 mM potassium phosphate buffer, pH 7.5, at 40°C Pyrococcus furiosus
1.5.99.B2 9.9
-
 L-proline wild type enzyme, in 100 mM potassium phosphate buffer, pH 7.5, at 60°C Pyrococcus furiosus
1.5.99.B2 14.46
-
 L-proline mutant enzyme H225Q, in 100 mM potassium phosphate buffer, pH 7.5, at 60°C Pyrococcus furiosus
1.5.99.B2 19.67
-
 L-proline mutant enzyme H225A, in 100 mM potassium phosphate buffer, pH 7.5, at 60°C Pyrococcus furiosus
1.5.99.B2 19.9
-
 L-proline wild type enzyme, in 100 mM potassium phosphate buffer, pH 7.5, at 70°C Pyrococcus furiosus
1.5.99.B2 30.8
-
 L-proline wild type enzyme, in 100 mM potassium phosphate buffer, pH 7.5, at 80°C Pyrococcus furiosus
1.5.99.B2 212.3
-
 L-pipecolic acid wild type enzyme, in 100 mM potassium phosphate buffer, pH 7.5, at 80°C Pyrococcus furiosus

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
1.5.99.B2 mitochondrion
-
 Pyrococcus furiosus 5739
-

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.5.99.B2 42440
-
 beta subunit, MALDI-TOF mass spectrometry Pyrococcus furiosus
1.5.99.B2 42470
-
 beta subunit, electrospray ionization mass spectrometry Pyrococcus furiosus

Organism

EC Number Organism UniProt Comment Textmining
1.5.99.B2 Pyrococcus furiosus
-
 strain DSM 3638
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.5.99.B2 DE52 anion exchange column chromatography Pyrococcus furiosus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.5.99.B2 L-pipecolic acid + acceptor + H2O
-
 Pyrococcus furiosus ? + reduced acceptor
-
 ?
1.5.99.B2 L-proline + acceptor + H2O preferred substrate Pyrococcus furiosus (S)-1-pyrroline-5-carboxylate + reduced acceptor
-
 ?
1.5.99.B2 additional information does not reduce NAD+ Pyrococcus furiosus ?
-
 ?
1.5.99.B2 sarcosine + acceptor + H2O
-
 Pyrococcus furiosus ? + reduced acceptor
-
 ?

Subunits

EC Number Subunits Comment Organism
1.5.99.B2 heterooctamer x-ray crystallography Pyrococcus furiosus

Synonyms

EC Number Synonyms Comment Organism
1.5.99.B2 L-proline dehydrogenase
-
 Pyrococcus furiosus
1.5.99.B2 PRODH
-
 Pyrococcus furiosus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.5.99.B2 100
-
 no loss of activity being evident up to 100°C, above this temperature, thermal denaturation of PRODH is apparent, with complete loss of activity after 10 min of incubation in glycerol buffer at temperatures above 115°C Pyrococcus furiosus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.5.99.B2 0.4
-
 L-pipecolic acid wild type enzyme in 100 mM potassium phosphate buffer, pH 7.5, at 80°C Pyrococcus furiosus
1.5.99.B2 0.4
-
 L-proline wild type enzyme, in 100 mM potassium phosphate buffer, pH 7.5, at 40°C Pyrococcus furiosus
1.5.99.B2 1.3
-
 L-proline wild type enzyme, in 100 mM potassium phosphate buffer, pH 7.5, at 50°C Pyrococcus furiosus
1.5.99.B2 1.4
-
 L-proline mutant enzyme H225A, in 100 mM potassium phosphate buffer, pH 7.5, at 60°C Pyrococcus furiosus
1.5.99.B2 4
-
 L-proline wild type enzyme, in 100 mM potassium phosphate buffer, pH 7.5, at 60°C Pyrococcus furiosus
1.5.99.B2 8.97
-
 L-proline mutant enzyme H225Q, in 100 mM potassium phosphate buffer, pH 7.5, at 60°C Pyrococcus furiosus
1.5.99.B2 10.4
-
 L-proline wild type enzyme, in 100 mM potassium phosphate buffer, pH 7.5, at 70°C Pyrococcus furiosus
1.5.99.B2 18.01
-
 L-proline wild type enzyme, in 100 mM potassium phosphate buffer, pH 7.5, at 80°C Pyrococcus furiosus
1.5.99.B2 37.17
-
 L-proline mutant enzyme Y215F, in 100 mM potassium phosphate buffer, pH 7.5, at 60°C Pyrococcus furiosus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.5.99.B2 5 10
-
 Pyrococcus furiosus

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
1.5.99.B2 5 10
-
 Pyrococcus furiosus

Cofactor

EC Number Cofactor Comment Organism Structure
1.5.99.B2 ATP the holoenzyme contains one ATP per alphabeta complex Pyrococcus furiosus
1.5.99.B2 FAD flavoprotein amine dehydrogenase, the holoenzyme contains one FAD per alphabeta complex Pyrococcus furiosus
1.5.99.B2 FMN the holoenzyme contains one FMN per alphabeta complex Pyrococcus furiosus