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Literature summary extracted from

  • Lawrence, C.C.; Sobey, W.J.; Field, R.A.; Baldwin, J.E.; Schofield, C.J.
    Purification and initial characterization of proline 4-hydroxylase from Streptomyces griseoviridis P8648: a 2-oxoacid, ferrous-dependent dioxygenase involved in etamycin biosynthesis (1996), Biochem. J., 313, 185-191.
No PubMed abstract available

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.14.11.2 catalase activation Streptomyces griseoviridis
1.14.11.2 pyridine 2,6-dicarboxylate activator Streptomyces griseoviridis
1.14.11.57 catalase presence of catalase causes 20-30% increase in the turnover of L-proline Streptomyces griseoviridis
1.14.11.57 pyridine-2,6-dicarboxylate
-
 Streptomyces griseoviridis

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.14.11.2 2-oxoglutarate at concentrations higher than 0.5 mM, decreases activity Streptomyces griseoviridis
1.14.11.2 3,4-dihydroxybenzoate
-
 Streptomyces griseoviridis
1.14.11.2 ascorbate the inhibition may result from competition for binding at the 2-oxoacid binding site between 2-oxoglutarate and L-ascorbate Streptomyces griseoviridis
1.14.11.2 Co2+ complete inhibition Streptomyces griseoviridis
1.14.11.2 Cu2+ causes a 20-30% fall in activity Streptomyces griseoviridis
1.14.11.2 diethyl dicarbonate 98% inhibition at 1 mM Streptomyces griseoviridis
1.14.11.2 Fe2+ inhibitory at a concentration higher than 0.5 mM Streptomyces griseoviridis
1.14.11.2 Hg2+ causes a 20-30% fall in activity Streptomyces griseoviridis
1.14.11.2 Mn2+ causes a 20-30% fall in activity Streptomyces griseoviridis
1.14.11.2 oxalylglycine
-
 Streptomyces griseoviridis
1.14.11.2 Pyridine 2,4-dicarboxylate
-
 Streptomyces griseoviridis
1.14.11.2 Pyridine 2,5-dicarboxylate
-
 Streptomyces griseoviridis
1.14.11.2 Zn2+ complete inhibition Streptomyces griseoviridis
1.14.11.57 2-oxoglutarate inhibitory above 0.5 mM Streptomyces griseoviridis
1.14.11.57 3,4-dihydroxybenzoate
-
 Streptomyces griseoviridis
1.14.11.57 Co2+ 0.5 mM, complete inhibition Streptomyces griseoviridis
1.14.11.57 diethyl dicarbonate complete inhibition. Inactivation can be partially prevented by the inclusion of L-proline and 2-oxoglutarate in the preincubation mixture Streptomyces griseoviridis
1.14.11.57 L-ascorbate ascorbate causes a time-dependent inhibition of L-proline hydroxylation. The addition of ascorbate does not stimulate L-proline-coupled turnover of 2-oxoglutarate, but does stimulate L-proline-uncoupled turnover Streptomyces griseoviridis
1.14.11.57 Pyridine-2,4-dicarboxylate competitive with respect to 2-oxoglutarate Streptomyces griseoviridis
1.14.11.57 Pyridine-2,5-dicarboxylate
-
 Streptomyces griseoviridis
1.14.11.57 Zn2+ 0.5 mM, complete inhibition Streptomyces griseoviridis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.11.2 0.032
-
 2-oxoglutarate
-
 Streptomyces griseoviridis
1.14.11.2 0.445
-
 L-proline
-
 Streptomyces griseoviridis
1.14.11.57 0.032
-
 2-oxoglutarate pH 7.5, 35°C Streptomyces griseoviridis
1.14.11.57 0.445
-
 L-proline pH 7.5, 35°C Streptomyces griseoviridis

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.14.11.2 Fe2+ the maximal rate of hydroxylation is attained at a ferrous ion concentration of approx. 0.03 mM and proceedes at approx. 8% of the maximal rate in the absence of exogenous iron, further suggesting the presence of residual enzyme-bound iron Streptomyces griseoviridis
1.14.11.2 Mn2+ no activation, causes a 20-30% fall in activity Streptomyces griseoviridis
1.14.11.57 Cu2+ about 10-15% of the activity with iron Streptomyces griseoviridis
1.14.11.57 Iron strict requirements for ferrous ion and 2-oxoglutarate, maximum activity at 0.03 mM Streptomyces griseoviridis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.14.11.2 35000
-
 1 * 35000, SDS-PAGE Streptomyces griseoviridis
1.14.11.2 37700
-
 gel filtration Streptomyces griseoviridis
1.14.11.57 35000
-
 1 * 35000, SDS-PAGE Streptomyces griseoviridis
1.14.11.57 38000
-
 gel filtration Streptomyces griseoviridis

Organism

EC Number Organism UniProt Comment Textmining
1.14.11.2 Streptomyces griseoviridis
-
-
-
1.14.11.57 Streptomyces griseoviridis
-
-
-
1.14.11.57 Streptomyces griseoviridis P8648
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.11.2 using chromatography on DEAE-Sepharose Fast flow column, ammonium sulfate precipitation, chromatography on phenyl-Superose column, second ammonium sulfate precipitation and column chromatography on Superdex G75 HR Streptomyces griseoviridis

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.14.11.57 907
-
 pH 7.5, 26°C Streptomyces griseoviridis

Storage Stability

EC Number Storage Stability Organism
1.14.11.57 -20°C, about 30% loss of activity by freezing and thawing Streptomyces griseoviridis
1.14.11.57 4°C, 4-5 h, stable Streptomyces griseoviridis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.11.2 L-proline + 2-oxoglutarate + O2 2-oxoglutarate is essential for hydroxylation Streptomyces griseoviridis 4-hydroxyproline + succinate + CO2
-
 ?
1.14.11.2 L-proline + 2-oxoglutarate + O2 2-oxoglutarate is essential for hydroxylation Streptomyces griseoviridis P8648 4-hydroxyproline + succinate + CO2
-
 ?
1.14.11.2 additional information 2-oxoglutarate cannot be replaced by oxalylglycine, 2-oxopentanoate, 2-oxoadipate, pyruvate or 2-oxomalonate Streptomyces griseoviridis ?
-
 ?
1.14.11.2 additional information 2-oxoglutarate cannot be replaced by oxalylglycine, 2-oxopentanoate, 2-oxoadipate, pyruvate or 2-oxomalonate Streptomyces griseoviridis P8648 ?
-
 ?
1.14.11.57 L-proline + 2-oxoglutarate + O2
-
 Streptomyces griseoviridis trans-4-hydroxy-L-proline + succinate + CO2
-
 ?
1.14.11.57 L-proline + 2-oxoglutarate + O2
-
 Streptomyces griseoviridis P8648 trans-4-hydroxy-L-proline + succinate + CO2
-
 ?
1.14.11.57 additional information the enzyme displays strict requirements for ferrous ion and 2-oxoglutarate, no cosubstrate: 2-oxopentanoate, 2-oxoadipate, pyruvate or 2-oxomalonate. L-Proline-uncoupled turnover of 2-oxoglutarate to succinate and CO2 is observed Streptomyces griseoviridis ?
-
 ?
1.14.11.57 additional information the enzyme displays strict requirements for ferrous ion and 2-oxoglutarate, no cosubstrate: 2-oxopentanoate, 2-oxoadipate, pyruvate or 2-oxomalonate. L-Proline-uncoupled turnover of 2-oxoglutarate to succinate and CO2 is observed Streptomyces griseoviridis P8648 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.14.11.2 monomer 1 * 35000, SDS-PAGE Streptomyces griseoviridis
1.14.11.57 monomer 1 * 35000, SDS-PAGE Streptomyces griseoviridis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.14.11.2 26
-
 assay at Streptomyces griseoviridis

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.14.11.2 0.0004
-
 oxalylglycine
-
 Streptomyces griseoviridis
1.14.11.2 0.005
-
 Pyridine 2,4-dicarboxylate
-
 Streptomyces griseoviridis
1.14.11.2 0.032
-
 3,4-dihydroxybenzoate
-
 Streptomyces griseoviridis
1.14.11.2 0.049
-
 Pyridine 2,5-dicarboxylate
-
 Streptomyces griseoviridis

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
1.14.11.57 0.005
-
 pH 7.5, 35°C Streptomyces griseoviridis Pyridine-2,4-dicarboxylate
1.14.11.57 0.032
-
 pH 7.5, 35°C Streptomyces griseoviridis 3,4-dihydroxybenzoate
1.14.11.57 0.049
-
 pH 7.5, 35°C Streptomyces griseoviridis Pyridine-2,5-dicarboxylate