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Literature summary for 1.5.99.13 extracted from

  • Satomura, T.; Kawakami, R.; Sakuraba, H.; Ohshima, T.
    Dye-linked D-proline dehydrogenase from hyperthermophilic archaeon Pyrobaculum islandicum is a novel FAD-dependent amino acid dehydrogenase (2002), J. Biol. Chem., 277, 12861-12867.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Pyrobaculum islandicum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.14
-
2,6-dichlorophenolindophenol pH 8.0, 50°C Pyrobaculum islandicum
4.2
-
D-proline pH 8.0, 50°C Pyrobaculum islandicum

Localization

Localization Comment Organism GeneOntology No. Textmining
membrane associated with Pyrobaculum islandicum 16020
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
40341
-
4 * 40341, calculated from sequence Pyrobaculum islandicum
42000
-
4 * 42000, SDS-PAGE Pyrobaculum islandicum
145000
-
gel filtration Pyrobaculum islandicum

Organism

Organism UniProt Comment Textmining
Pyrobaculum islandicum A1RVM8 JCM 9189
-
Pyrobaculum islandicum JCM 9189 A1RVM8 JCM 9189
-

Purification (Commentary)

Purification (Comment) Organism
-
Pyrobaculum islandicum

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
5.3
-
enzyme from Pyrobaculum islandicum Pyrobaculum islandicum
13.2
-
recombinant enzyme Pyrobaculum islandicum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
allo-4-hydroxy-D-proline + oxidized 2,6-dichloroindophenol + H2O 89% of the activity with D-proline Pyrobaculum islandicum ? + reduced 2,6-dichloroindophenol
-
?
allo-4-hydroxy-D-proline + oxidized 2,6-dichloroindophenol + H2O 89% of the activity with D-proline Pyrobaculum islandicum JCM 9189 ? + reduced 2,6-dichloroindophenol
-
?
D-alanine + oxidized 2,6-dichloroindophenol + H2O 26% of the activity with D-proline Pyrobaculum islandicum pyruvate + NH3 + reduced 2,6-dichloroindophenol
-
?
D-alanine + oxidized 2,6-dichloroindophenol + H2O 26% of the activity with D-proline Pyrobaculum islandicum JCM 9189 pyruvate + NH3 + reduced 2,6-dichloroindophenol
-
?
D-arginine + oxidized 2,6-dichloroindophenol + H2O 10% of the activity with D-proline Pyrobaculum islandicum 2-oxo-5-guanidinopentanoate + NH3 + reduced 2,6-dichloroindophenol
-
?
D-arginine + oxidized 2,6-dichloroindophenol + H2O 10% of the activity with D-proline Pyrobaculum islandicum JCM 9189 2-oxo-5-guanidinopentanoate + NH3 + reduced 2,6-dichloroindophenol
-
?
D-aspartate + oxidized 2,6-dichloroindophenol + H2O 21% of the activity with D-proline Pyrobaculum islandicum oxaloacetate + NH3 + reduced 2,6-dichloroindophenol
-
?
D-aspartate + oxidized 2,6-dichloroindophenol + H2O 21% of the activity with D-proline Pyrobaculum islandicum JCM 9189 oxaloacetate + NH3 + reduced 2,6-dichloroindophenol
-
?
D-glutamate + oxidized 2,6-dichloroindophenol + H2O 23% of the activity with D-proline Pyrobaculum islandicum 2-oxo-glutarate + NH3 + reduced 2,6-dichloroindophenol
-
?
D-histidine + oxidized 2,6-dichloroindophenol + H2O 30% of the activity with D-proline Pyrobaculum islandicum 3-(1H-imidazol-4-yl)-2-oxopropanoate + NH3 + reduced 2,6-dichloroindophenol
-
?
D-isoleucine + oxidized 2,6-dichloroindophenol + H2O 49% of the activity with D-proline Pyrobaculum islandicum 3-methyl-2-oxopropanoate + NH3 + reduced 2,6-dichloroindophenol
-
?
D-leucine + oxidized 2,6-dichloroindophenol + H2O 39% of the activity with D-proline Pyrobaculum islandicum 4-methyl-2-oxopentanoate + NH3 + reduced 2,6-dichloroindophenol
-
?
D-phenylalanine + oxidized 2,6-dichloroindophenol + H2O 28% of the activity with D-proline Pyrobaculum islandicum phenylpyruvate + NH3 + reduced 2,6-dichloroindophenol
-
?
D-proline + 2,6-dichlorophenolindophenol
-
Pyrobaculum islandicum 1-pyrroline-2-carboxylate + reduced 2,6-dichlorophenolindophenol
-
r
D-proline + oxidized 2,6-dichloroindophenol D-proline is the most preferred substrate among the D-amino acids. 2,6-Dichloroindophenol is the most preferred electron acceptor Pyrobaculum islandicum DELTA1-pyrroline 2-carboxylate + reduced 2,6-dichloroindophenol
-
?
D-proline + oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide 33% of the activity with oxidized 2,6-dichloroindophenol Pyrobaculum islandicum DELTA1-pyrroline 2-carboxylate + reduced 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
?
D-proline + oxidized phenazine methosulfate 43% of the activity with oxidized 2,6-dichloroindophenol Pyrobaculum islandicum DELTA1-pyrroline 2-carboxylate + reduced phenazine methosulfate
-
?
D-serine + oxidized 2,6-dichloroindophenol + H2O 13% of the activity with D-proline Pyrobaculum islandicum 3-hydroxy-2-oxopropanoate + NH3 + reduced 2,6-dichloroindophenol
-
?
D-threonine + oxidized 2,6-dichloroindophenol + H2O 15% of the activity with D-proline Pyrobaculum islandicum 3-hydroxy-2-oxobutanoate + NH3 + reduced 2,6-dichloroindophenol
-
?
D-tryptophan + oxidized 2,6-dichloroindophenol + H2O 14% of the activity with D-proline Pyrobaculum islandicum 3-indole-2-oxopropanoate + NH3 + reduced 2,6-dichloroindophenol
-
?
D-valine + oxidized 2,6-dichloroindophenol + H2O 41% of the activity with D-proline Pyrobaculum islandicum 2-oxoisopentanoate + NH3 + reduced 2,6-dichloroindophenol
-
?
additional information ferricyanide, NAD+, NADP+, O2, menadione (vitamin K3), benzyl viologen, horse heart cytochrome c, and methylene blue are inert as electron acceptors Pyrobaculum islandicum ?
-
?
additional information ferricyanide, NAD+, NADP+, O2, menadione (vitamin K3), benzyl viologen, horse heart cytochrome c, and methylene blue are inert as electron acceptors Pyrobaculum islandicum JCM 9189 ?
-
?

Subunits

Subunits Comment Organism
tetramer 4 * 42000, SDS-PAGE Pyrobaculum islandicum
tetramer 4 * 40341, calculated from sequence Pyrobaculum islandicum

Synonyms

Synonyms Comment Organism
dye-linked D-proline dehydrogenase
-
Pyrobaculum islandicum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
70
-
the activity is linearly increased with increasing temperature to 70°C. The highest activity is above 70°C, and the correct assay is not achieved above 70°C, because of the non-enzymatic decolorization of 2,6-dichloroindophenol Pyrobaculum islandicum

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
80
-
10 min, enzyme retains its full activity Pyrobaculum islandicum
100
-
10 min, 90% loss of activity Pyrobaculum islandicum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
D-proline dehydrogenation Pyrobaculum islandicum

pH Range

pH Minimum pH Maximum Comment Organism
6.5 8.5 50% of maximal activity at pH 6.5 and at pH 8.5, proline dehydrogenation Pyrobaculum islandicum

pH Stability

pH Stability pH Stability Maximum Comment Organism
4 10 50°C, 10 min, enzyme retains its full activity Pyrobaculum islandicum

Cofactor

Cofactor Comment Organism Structure
FAD prosthetic group Pyrobaculum islandicum