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Literature summary for 1.5.1.2 extracted from

  • Yang, Y.; Xu, S.; Zhang, M.; Jin, R.; Zhang, L.; Bao, J.; Wang, H.
    Purification and characterization of a functionally active Mycobacterium tuberculosis pyrroline-5-carboxylate reductase (2006), Protein Expr. Purif., 45, 241-248.
    View publication on PubMed

Application

Application Comment Organism
medicine pyrroline-5-carboxylate reductase is related to virulence of Mycobacterium tuberculosis Mycobacterium tuberculosis

Cloned(Commentary)

Cloned (Comment) Organism
into the pET30a vector for expression in Escherichia coli BL21DE3 Mycobacterium tuberculosis

Inhibitors

Inhibitors Comment Organism Structure
deoxycholate
-
Mycobacterium tuberculosis
H2O2
-
Mycobacterium tuberculosis
methanol
-
Mycobacterium tuberculosis
SDS
-
Mycobacterium tuberculosis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.04
-
1-pyrroline-5-carboxylate cofactor NADH Mycobacterium tuberculosis
0.047
-
1-pyrroline-5-carboxylate cofactor NADPH Mycobacterium tuberculosis
0.057
-
NADH
-
Mycobacterium tuberculosis
0.059
-
NADPH
-
Mycobacterium tuberculosis
0.093
-
NADP+
-
Mycobacterium tuberculosis
0.12
-
L-proline
-
Mycobacterium tuberculosis

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ activity is significantly increased in the forward reaction, optimal concentration of the cation is about 50-100 microM Mycobacterium tuberculosis
Mn2+ activity is significantly increased in the forward reaction, optimal concentration of the cation is about 50-100 microM Mycobacterium tuberculosis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
32000
-
after enterokinase treatment, estimated by SDS-PAGE Mycobacterium tuberculosis
365000
-
decamer, analysis of nondegenerated protein by native gel electrophoresis Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1-pyrroline-5-carboxylate + NADPH + H+ Mycobacterium tuberculosis
-
L-proline + NADP+
-
r

Organism

Organism UniProt Comment Textmining
Mycobacterium tuberculosis
-
-
-

Purification (Commentary)

Purification (Comment) Organism
by Ni-NTA His-binding resin affinity chromatography, the N-terminal fusion partner is cleaved by enterokinase Mycobacterium tuberculosis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.9
-
purification step supernatant, reverse reaction Mycobacterium tuberculosis
1.7
-
purification step supernatant, forward reaction Mycobacterium tuberculosis
30
-
purification step Ni-NTA column, reverse reaction Mycobacterium tuberculosis
57
-
purification step Ni-NTA column, forward reaction Mycobacterium tuberculosis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1-pyrroline-5-carboxylate + NADH + H+
-
Mycobacterium tuberculosis L-proline + NAD+
-
r
1-pyrroline-5-carboxylate + NADPH + H+
-
Mycobacterium tuberculosis L-proline + NADP+
-
r
1-pyrroline-5-carboxylate + NADPH + H+ activity assay, forward reaction Mycobacterium tuberculosis L-proline + NADP+
-
r

Subunits

Subunits Comment Organism
decamer 10*32000-35600, estimated by SDS-PAGE, confirmed by spectrometric analysis Mycobacterium tuberculosis

Synonyms

Synonyms Comment Organism
P5CR
-
Mycobacterium tuberculosis
pyrroline-5-carboxylate reductase
-
Mycobacterium tuberculosis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
activity assay, forward and reverse reaction Mycobacterium tuberculosis

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
40 50
-
Mycobacterium tuberculosis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
60
-
half life of the His-tagged protein is about 20 min Mycobacterium tuberculosis
70
-
His-tagged protein is rapidly inactivated Mycobacterium tuberculosis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.00983
-
1-pyrroline-5-carboxylate cofactor NADPH Mycobacterium tuberculosis
137.9
-
NADP+
-
Mycobacterium tuberculosis
212.8
-
L-proline
-
Mycobacterium tuberculosis
310.2
-
NADH
-
Mycobacterium tuberculosis
321.6
-
NADPH
-
Mycobacterium tuberculosis
387.3
-
1-pyrroline-5-carboxylate cofactor NADH Mycobacterium tuberculosis
451.7
-
1-pyrroline-5-carboxylate cofactor NADPH Mycobacterium tuberculosis
4000
-
NADPH
-
Mycobacterium tuberculosis
4170
-
NADH
-
Mycobacterium tuberculosis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
activity assay, forward reaction Mycobacterium tuberculosis
10.3
-
activity assay, reverse reaction Mycobacterium tuberculosis

Cofactor

Cofactor Comment Organism Structure
NADPH
-
Mycobacterium tuberculosis