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Literature summary for 1.5.99.B2 extracted from

  • Satomura, T.; Zhang, X.D.; Hara, Y.; Doi, K.; Sakuraba, H.; Ohshima, T.
    Characterization of a novel dye-linked L-proline dehydrogenase from an aerobic hyperthermophilic archaeon, Pyrobaculum calidifontis (2011), Appl. Microbiol. Biotechnol., 89, 1075-1082.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) Codon Plus RIL cells Pyrobaculum calidifontis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.026
-
2,6-dichlorophenolindophenol with L-proline as cosubstrate, in 200 mM Tris-HCl buffer (pH 8.0), at 50°C Pyrobaculum calidifontis
0.035
-
2,6-dichlorophenolindophenol with L-hydroxyproline as cosubstrate, in 200 mM Tris-HCl buffer (pH 8.0), at 50°C Pyrobaculum calidifontis
1.67
-
L-proline in 200 mM Tris-HCl buffer (pH 8.0), at 50°C Pyrobaculum calidifontis
6.05
-
L-hydroxyproline in 200 mM Tris-HCl buffer (pH 8.0), at 50°C Pyrobaculum calidifontis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
46000
-
2 * 46000, SDS-PAGE Pyrobaculum calidifontis
46259
-
2 * 46259, calculated from amino acid sequence Pyrobaculum calidifontis
108000
-
gel filtration Pyrobaculum calidifontis

Organism

Organism UniProt Comment Textmining
Pyrobaculum calidifontis
-
-
-
Pyrobaculum calidifontis JCM 11548
-
-
-

Purification (Commentary)

Purification (Comment) Organism
DEAE Toyopearl 650M column chromatography and Superdex 200 gel filtration Pyrobaculum calidifontis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.026
-
crude extract, pH 8.0, at 50°C Pyrobaculum calidifontis
4.25
-
after 163fold purification, pH 8.0, at 50°C Pyrobaculum calidifontis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-hydroxyproline + 2,6-dichlorophenolindophenol + H2O 72% activity compared to L-proline Pyrobaculum calidifontis ? + reduced 2,6-dichlorophenolindophenol
-
?
L-hydroxyproline + 2,6-dichlorophenolindophenol + H2O 72% activity compared to L-proline Pyrobaculum calidifontis JCM 11548 ? + reduced 2,6-dichlorophenolindophenol
-
?
L-proline + 2,6-dichlorophenolindophenol + H2O L-proline is the most preferred substrate and 2,6-dichlorophenolindophenol is the most preferred electron acceptor Pyrobaculum calidifontis DELTA1-pyrroline-5-carboxylate + reduced 2,6-dichlorophenolindophenol
-
?
L-proline + 2,6-dichlorophenolindophenol + H2O L-proline is the most preferred substrate and 2,6-dichlorophenolindophenol is the most preferred electron acceptor Pyrobaculum calidifontis JCM 11548 DELTA1-pyrroline-5-carboxylate + reduced 2,6-dichlorophenolindophenol
-
?
L-proline + 4-iodonitrotetrazolium violet + H2O 9% activity compared to 2,6-dichlorophenolindophenol Pyrobaculum calidifontis DELTA1-pyrroline-5-carboxylate + reduced 4-iodonitrotetrazolium violet
-
?
L-proline + 4-iodonitrotetrazolium violet + H2O 9% activity compared to 2,6-dichlorophenolindophenol Pyrobaculum calidifontis JCM 11548 DELTA1-pyrroline-5-carboxylate + reduced 4-iodonitrotetrazolium violet
-
?
L-proline + ferricyanide 93% activity compared to 2,6-dichlorophenolindophenol Pyrobaculum calidifontis DELTA1-pyrroline-5-carboxylate + ferrocyanide
-
?
L-proline + ferricyanide 93% activity compared to 2,6-dichlorophenolindophenol Pyrobaculum calidifontis JCM 11548 DELTA1-pyrroline-5-carboxylate + ferrocyanide
-
?
additional information sarcosine, D-proline, L-ornithine, L-glutamate, L-arginine, L-serine, L-leucine, L-valine, and L-alanine are inert as substrates. NAD+, NADP+, and bovine heart cytochrome c are inert as electron acceptors Pyrobaculum calidifontis ?
-
?
additional information sarcosine, D-proline, L-ornithine, L-glutamate, L-arginine, L-serine, L-leucine, L-valine, and L-alanine are inert as substrates. NAD+, NADP+, and bovine heart cytochrome c are inert as electron acceptors Pyrobaculum calidifontis JCM 11548 ?
-
?

Subunits

Subunits Comment Organism
homodimer 2 * 46000, SDS-PAGE Pyrobaculum calidifontis
homodimer 2 * 46259, calculated from amino acid sequence Pyrobaculum calidifontis

Synonyms

Synonyms Comment Organism
L-proDH
-
Pyrobaculum calidifontis
L-proline dehydrogenase
-
Pyrobaculum calidifontis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
80
-
the optimum temperature for enzyme activity is above 80°C Pyrobaculum calidifontis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
100 105 the enzyme retains more than 90% of its activity after incubation at 100°C for 120 min (pH 7.5). The half-life at 105°C is about 90 min Pyrobaculum calidifontis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
-
Pyrobaculum calidifontis

pH Range

pH Minimum pH Maximum Comment Organism
6 9 half of the activity is retained at pH 6.0 and 9.0 Pyrobaculum calidifontis

pH Stability

pH Stability pH Stability Maximum Comment Organism
4.5 9 the enzyme retains more than 90% of its activity after incubation at pH values 4.5-9.0 for 30 min at 50°C Pyrobaculum calidifontis

Cofactor

Cofactor Comment Organism Structure
FAD
-
Pyrobaculum calidifontis