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Literature summary extracted from

  • Huijbers, M.M.; van Berkel, W.J.
    High yields of active Thermus thermophilus proline dehydrogenase are obtained using maltose-binding protein as a solubility tag (2015), Biotechnol. J., 10, 395-403 .
    View publication on PubMed

Application

EC Number Application Comment Organism
1.5.5.2 synthesis ProDH is of interest for practical applications because the proline imino acid can serve as a building block for a wide range of peptides and antibiotics Thermus thermophilus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.5.5.2 gene TT_C1214, recombinant expression of functional and soluble MBP-tagged enzyme in Escherichia coli TOP10 cells. The MBP tag inhibits the self-association of TtProDH, preventing to a large extent the formation of insoluble protein aggregates Thermus thermophilus

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.5.5.2 GuHCl enzyme unfolding at 1 M, 0.5 M is not enough for proper unfolding. The fusion protein forms visible aggregates due to unfolding of MBP Thermus thermophilus

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
1.5.5.2 mitochondrial membrane a membrane-associated protein Thermus thermophilus 31966
-

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.5.5.2 additional information
-
 the MBP tag inhibits the self-association of TtProDH, preventing to a large extent the formation of insoluble protein aggregates Thermus thermophilus
1.5.5.2 78675
-
 recombinant MBP-tagged enzyme, sequence calculation Thermus thermophilus
1.5.5.2 78677
-
 recombinant MBP-tagged enzyme, mass spectrometry Thermus thermophilus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.5.5.2 L-proline + a quinone Thermus thermophilus
-
 (S)-1-pyrroline-5-carboxylate + a quinol
-
 ?
1.5.5.2 L-proline + a quinone Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039
-
 (S)-1-pyrroline-5-carboxylate + a quinol
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.5.5.2 Thermus thermophilus Q72IB8
-
-
1.5.5.2 Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039 Q72IB8
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.5.5.2 recombinant MBP-tagged enzyme from Escherichia coli, gel filtration, and tag cleavage by trypsin, followed by ultrafiltration Thermus thermophilus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.5.5.2 4
-
 purified recombinant MBP-tagged enzyme, pH 8.0, 25°C Thermus thermophilus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.5.5.2 L-proline + 2,6-dichlorphenol-indophenol
-
 Thermus thermophilus (S)-1-pyrroline-5-carboxylate + reduced 2,6-dichlorphenol-indophenol
-
 ?
1.5.5.2 L-proline + 2,6-dichlorphenol-indophenol
-
 Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039 (S)-1-pyrroline-5-carboxylate + reduced 2,6-dichlorphenol-indophenol
-
 ?
1.5.5.2 L-proline + a quinone
-
 Thermus thermophilus (S)-1-pyrroline-5-carboxylate + a quinol
-
 ?
1.5.5.2 L-proline + a quinone a FAD-dependent reaction Thermus thermophilus (S)-1-pyrroline-5-carboxylate + a quinol
-
 ?
1.5.5.2 L-proline + a quinone
-
 Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039 (S)-1-pyrroline-5-carboxylate + a quinol
-
 ?
1.5.5.2 L-proline + a quinone a FAD-dependent reaction Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039 (S)-1-pyrroline-5-carboxylate + a quinol
-
 ?

Subunits

EC Number Subunits Comment Organism
1.5.5.2 More recombinant wild-type detagged enzyme and recombinant wild-type MBP-tagged enzyme both form oligomers. Peptide mapping Thermus thermophilus

Synonyms

EC Number Synonyms Comment Organism
1.5.5.2 PRODH
-
 Thermus thermophilus
1.5.5.2 TtProDH
-
 Thermus thermophilus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.5.5.2 25
-
 assay at Thermus thermophilus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.5.5.2 additional information
-
 enzyme unfolding experiments using recombinant MBP-tagged enzyme in 20 mM Hepes, pH 8.0, at 60-95°C, the MBP-tagged enzyme unfolds at lower temperature around 56°C due to unfolding of the MBP tag Thermus thermophilus
1.5.5.2 90
-
 with the native enzyme, rapid loss of activity only occurs at temperatures above 90°C Thermus thermophilus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.5.5.2 8
-
 assay at Thermus thermophilus

Cofactor

EC Number Cofactor Comment Organism Structure
1.5.5.2 FAD enzyme-bound, dependent on. Addition of excess FAD hardly improves the enzyme activity, confirming that the enzyme is fully saturated with FAD Thermus thermophilus

General Information

EC Number General Information Comment Organism
1.5.5.2 metabolism proline dehydrogenase (ProDH) catalyzes the FAD-dependent oxidation of proline to DELTA1-pyrroline-5-carboxylate, the first step of proline catabolism in many organisms Thermus thermophilus