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Literature summary extracted from

  • Serrano, H.; Blanchard, J.S.
    Kinetic and isotopic characterization of L-proline dehydrogenase from Mycobacterium tuberculosis (2013), Biochemistry, 52, 5009-5015 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.5.99.B2 recombinant overexpression of C-terminally His6-tagged wild-type and mutant enzymes in Mycobacterium smegmatis mc24517 cells using vector pYUB1062 Mycobacterium tuberculosis

Protein Variants

EC Number Protein Variants Comment Organism
1.5.99.B2 K110A site-directed mutagenesis, inactive mutant Mycobacterium tuberculosis
1.5.99.B2 Y203F site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Mycobacterium tuberculosis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.5.99.B2 additional information
-
 additional information Michaelis-Menten kinetics, steady-state kinetics and pre-steady-state kinetics of wild-type and mutant enzymes, stopped-flow kinetic isotope effects, bisubstrate kinetic analysis, overview Mycobacterium tuberculosis
1.5.99.B2 3.4
-
 2,6-dichlorophenolindophenol pH 7.1, 25°C, recombinant wild-type enzyme Mycobacterium tuberculosis
1.5.99.B2 5.7
-
 L-proline pH 7.1, 25°C, recombinant wild-type enzyme Mycobacterium tuberculosis
1.5.99.B2 860
-
 L-proline pH 7.1, 25°C, recombinant mutant Y203F Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.5.99.B2 L-proline + acceptor Mycobacterium tuberculosis
-
 (S)-1-pyrroline-5-carboxylate + reduced acceptor
-
 ?
1.5.99.B2 L-proline + acceptor Mycobacterium tuberculosis H37Rv
-
 (S)-1-pyrroline-5-carboxylate + reduced acceptor
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.5.99.B2 Mycobacterium tuberculosis O50444
-
-
1.5.99.B2 Mycobacterium tuberculosis H37Rv O50444
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.5.99.B2 recombinant C-terminally His6-tagged wild-type and mutant enzymes from Mycobacterium smegmatis mc24517 cells by nickel affinity chromatography Mycobacterium tuberculosis

Reaction

EC Number Reaction Comment Organism Reaction ID
1.5.99.B2 L-proline + acceptor = (S)-1-pyrroline-5-carboxylate + reduced acceptor ping-pong mechanism, the transfer of hydride from L-proline to FAD is rate-limiting for the reductive half-reaction, but the overall reaction is limited by reduced flavin reoxidation in the second half-reaction. Solvent and multiple kinetic isotope effects suggest that L-proline oxidation occurs in a stepwise rather than concerted mechanism Mycobacterium tuberculosis
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.5.99.B2 L-proline + 2,6-dichlorophenolindophenol artificial electron acceptor Mycobacterium tuberculosis (S)-1-pyrroline-5-carboxylate + reduced 2,6-dichlorophenolindophenol
-
 ?
1.5.99.B2 L-proline + 2,6-dichlorophenolindophenol artificial electron acceptor Mycobacterium tuberculosis H37Rv (S)-1-pyrroline-5-carboxylate + reduced 2,6-dichlorophenolindophenol
-
 ?
1.5.99.B2 L-proline + acceptor
-
 Mycobacterium tuberculosis (S)-1-pyrroline-5-carboxylate + reduced acceptor
-
 ?
1.5.99.B2 L-proline + acceptor
-
 Mycobacterium tuberculosis H37Rv (S)-1-pyrroline-5-carboxylate + reduced acceptor
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.5.99.B2 PRODH
-
 Mycobacterium tuberculosis
1.5.99.B2 proline dehydrogenase
-
 Mycobacterium tuberculosis
1.5.99.B2 prub
-
 Mycobacterium tuberculosis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.5.99.B2 25
-
 assay at Mycobacterium tuberculosis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.5.99.B2 33.5
-
 L-proline pH 7.1, 25°C, recombinant wild-type enzyme Mycobacterium tuberculosis
1.5.99.B2 100.6
-
 L-proline pH 7.1, 25°C, recombinant mutant Y203F Mycobacterium tuberculosis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.5.99.B2 7.1 7.4 assay at Mycobacterium tuberculosis

Cofactor

EC Number Cofactor Comment Organism Structure
1.5.99.B2 FAD
-
 Mycobacterium tuberculosis

General Information

EC Number General Information Comment Organism
1.5.99.B2 metabolism proline dehydrogenase (ProDH) is a ubiquitous flavoenzyme involved in the biosynthesis of L-glutamate Mycobacterium tuberculosis
1.5.99.B2 additional information residue Y203 serves to bind and orient L-proline in the active site, residue K110 is the active site base observed in the pH profile Mycobacterium tuberculosis
1.5.99.B2 physiological function the monofunctional proline dehydrogenase (ProDH) performs the flavin-dependent oxidation of L-proline to DELTA1-pyrroline-5-carboxylate in the proline catabolic pathway Mycobacterium tuberculosis

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.5.99.B2 0.12
-
 L-proline pH 7.1, 25°C, recombinant mutant Y203F Mycobacterium tuberculosis
1.5.99.B2 5.88
-
 L-proline pH 7.1, 25°C, recombinant wild-type enzyme Mycobacterium tuberculosis