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Literature summary for 1.5.99.B2 extracted from
- Kinetic and isotopic characterization of L-proline dehydrogenase from Mycobacterium tuberculosis (2013), Biochemistry, 52, 5009-5015 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| recombinant overexpression of C-terminally His6-tagged wild-type and mutant enzymes in Mycobacterium smegmatis mc24517 cells using vector pYUB1062 | Mycobacterium tuberculosis |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| K110A | site-directed mutagenesis, inactive mutant | Mycobacterium tuberculosis |
| Y203F | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Mycobacterium tuberculosis |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 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 | |
| 3.4 | - |
2,6-dichlorophenolindophenol | pH 7.1, 25°C, recombinant wild-type enzyme | Mycobacterium tuberculosis |  |
| 5.7 | - |
L-proline | pH 7.1, 25°C, recombinant wild-type enzyme | Mycobacterium tuberculosis | |
| 860 | - |
L-proline | pH 7.1, 25°C, recombinant mutant Y203F | Mycobacterium tuberculosis | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| L-proline + acceptor | Mycobacterium tuberculosis | - |
(S)-1-pyrroline-5-carboxylate + reduced acceptor | - |
? | |
| L-proline + acceptor | Mycobacterium tuberculosis H37Rv | - |
(S)-1-pyrroline-5-carboxylate + reduced acceptor | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Mycobacterium tuberculosis | O50444 | - |
- |
| Mycobacterium tuberculosis H37Rv | O50444 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant C-terminally His6-tagged wild-type and mutant enzymes from Mycobacterium smegmatis mc24517 cells by nickel affinity chromatography | Mycobacterium tuberculosis |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| 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 |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| L-proline + 2,6-dichlorophenolindophenol | artificial electron acceptor | Mycobacterium tuberculosis | (S)-1-pyrroline-5-carboxylate + reduced 2,6-dichlorophenolindophenol | - |
? | |
| L-proline + 2,6-dichlorophenolindophenol | artificial electron acceptor | Mycobacterium tuberculosis H37Rv | (S)-1-pyrroline-5-carboxylate + reduced 2,6-dichlorophenolindophenol | - |
? | |
| L-proline + acceptor | - |
Mycobacterium tuberculosis | (S)-1-pyrroline-5-carboxylate + reduced acceptor | - |
? | |
| L-proline + acceptor | - |
Mycobacterium tuberculosis H37Rv | (S)-1-pyrroline-5-carboxylate + reduced acceptor | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| PRODH | - |
Mycobacterium tuberculosis |
| proline dehydrogenase | - |
Mycobacterium tuberculosis |
| prub | - |
Mycobacterium tuberculosis |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | - |
assay at | Mycobacterium tuberculosis |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 33.5 | - |
L-proline | pH 7.1, 25°C, recombinant wild-type enzyme | Mycobacterium tuberculosis | |
| 100.6 | - |
L-proline | pH 7.1, 25°C, recombinant mutant Y203F | Mycobacterium tuberculosis | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.1 | 7.4 | assay at | Mycobacterium tuberculosis |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| FAD | - |
Mycobacterium tuberculosis | |
General Information
| General Information | Comment | Organism |
|---|---|---|
| metabolism | proline dehydrogenase (ProDH) is a ubiquitous flavoenzyme involved in the biosynthesis of L-glutamate | Mycobacterium tuberculosis |
| 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 |
| 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]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.12 | - |
L-proline | pH 7.1, 25°C, recombinant mutant Y203F | Mycobacterium tuberculosis | |
| 5.88 | - |
L-proline | pH 7.1, 25°C, recombinant wild-type enzyme | Mycobacterium tuberculosis | |
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