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Literature summary for 6.3.3.6 extracted from

  • Raber, M.L.; Arnett, S.O.; Townsend, C.A.
    A conserved tyrosyl-glutamyl catalytic dyad in evolutionarily linked enzymes: carbapenam synthetase and beta-lactam synthetase (2009), Biochemistry, 48, 4959-40971.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 6.3.3.6

Protein Variants

Protein Variants Comment Organism
E380A kcat/Km is 74.3fold lower compared to wild-type value Pectobacterium carotovorum
E380D retains a functional general base with a pKa of about 7.4 for kcat. The acidic region of the log (kcat/Km) versus pH profiles displays an ionizable group with a pKa of about 7.7. kcat/Km is 1.4fold lower compared to wild-type value Pectobacterium carotovorum
E380Q variant displays a plateau in the acidic region. The acidic region of the log (kcat/Km) versus pH profiles becomes pH-independent for E380Q. kcat/Km is 3.8fold lower compared to wild-type value Pectobacterium carotovorum
Y345A mutant exhibits a 5fold increase in Km of (2S,5S)-5-carboxymethylproline and a 165fold decrease in specificity constant compared to wild-type enzyme. kcat/Km is 165fold lower compared to wild-type value Pectobacterium carotovorum
Y345A/E380A kcat/Km is 87fold lower compared to wild-type value Pectobacterium carotovorum
Y345F pH-dependent kcat and kcat/Km plots retain the bell-shaped curve of wild-type CPS with similar pK values. kcat/Km is 1.7fold lower compared to wild-type value Pectobacterium carotovorum
Y345F/E380D kcat/Km is 87fold lower compared to wild-type value Pectobacterium carotovorum
Y345F/E380Q kcat/Km is 95fold lower compared to wild-type value Pectobacterium carotovorum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.075
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, mutant enzyme E380Q Pectobacterium carotovorum
0.27
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, mutant enzyme Y345F Pectobacterium carotovorum
0.35
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, wild-type enzyme Pectobacterium carotovorum
0.36
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, mutant enzyme E380D Pectobacterium carotovorum
0.4
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, mutant enzyme Y345F/E380A Pectobacterium carotovorum
0.49
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, mutant enzyme E380A Pectobacterium carotovorum
0.49
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, mutant enzyme Y345F/E380D Pectobacterium carotovorum
0.9
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, mutant enzyme Y345F/E380Q Pectobacterium carotovorum
1.7
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, mutant enzyme Y345A Pectobacterium carotovorum

Organism

Organism UniProt Comment Textmining
Pectobacterium carotovorum
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + (2S,5S)-5-carboxymethylproline catalytic Tyr-Glu dyad is demonstrated by site-directed mutagenesis and kinetic experiments that compare the wild-type enzymes to their respective mutant proteins using pH–rate profiles, 32P-incorporation experiments, solvent isotope effects, proton inventory, and viscosity variation Pectobacterium carotovorum AMP + diphosphate + (3S,5S)-carbapenam 3-carboxylate
-
 ?

Synonyms

Synonyms Comment Organism
CPS
-
 Pectobacterium carotovorum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Pectobacterium carotovorum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0051
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, mutant enzyme Y345F/E380A Pectobacterium carotovorum
0.0059
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, mutant enzyme Y345F/E380D Pectobacterium carotovorum
0.0067
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, mutant enzyme E380A Pectobacterium carotovorum
0.011
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, mutant enzyme Y345A Pectobacterium carotovorum
0.011
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, mutant enzyme Y345F/E380Q Pectobacterium carotovorum
0.02
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, mutant enzyme E380Q Pectobacterium carotovorum
0.17
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, mutant enzyme Y345F Pectobacterium carotovorum
0.26
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, mutant enzyme E380D Pectobacterium carotovorum
0.36
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, wild-type enzyme Pectobacterium carotovorum

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.0063
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, mutant enzyme Y345A Pectobacterium carotovorum
0.011
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, mutant enzyme Y345F/E380Q Pectobacterium carotovorum
0.012
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, mutant enzyme Y345F/E380A Pectobacterium carotovorum
0.012
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, mutant enzyme Y345F/E380D Pectobacterium carotovorum
0.014
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, mutant enzyme E380A Pectobacterium carotovorum
0.27
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, mutant enzyme E380Q Pectobacterium carotovorum
0.63
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, mutant enzyme Y345F Pectobacterium carotovorum
0.72
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, mutant enzyme E380D Pectobacterium carotovorum
1.04
-
 (2S,5S)-5-carboxymethylproline pH 8.0, 25°C, wild-type enzyme Pectobacterium carotovorum