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Literature summary extracted from

  • Chen, H.; Gollnick, P.; Phillips, R.S.
    Site-directed mutagenesis of His343-->Ala in Citrobacter freundii tyrosine phenol-lyase. Effects on the kinetic mechanism and rate-determining step (1995), Eur. J. Biochem., 229, 540-549.
    View publication on PubMed

Protein Variants

EC Number Protein Variants Comment Organism
4.1.99.2 H343A all substrates for alpha,beta-elimination, except S-ethyl-L-Cys, exhibit lower turnover number values with the mutant enzyme than with the wild-type enzyme. The mutant shows slower rates of deuterium isotope exchange for L-Phe and L-Met than does the wild type enzyme. The turnover-number for 3-fluoro-L-Tyr is pH-dependent for the mutant enzyme, whereas it is pH-independent for the wild-type enzyme. His343 does play an important function in catalysis, possibly by facilitating the conformational change from an ‘open' to ‘closed' form when substrates bind Citrobacter freundii

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.1.99.2 0.1
-
 S-(o-nitrophenyl)-L-Cys wild-type enzyme Citrobacter freundii
4.1.99.2 0.1
-
 3-fluoro-L-Tyr wild-type enzyme Citrobacter freundii
4.1.99.2 0.2
-
 L-Tyr wild-type enzyme Citrobacter freundii
4.1.99.2 0.2
-
 S-benzyl-L-Cys wild-type enzyme Citrobacter freundii
4.1.99.2 0.5
-
 S-(o-nitrophenyl)-L-Cys mutant enzyme H343A Citrobacter freundii
4.1.99.2 1.2
-
 S-ethyl-L-Cys mutant enzyme H343A Citrobacter freundii
4.1.99.2 1.2
-
 S-benzyl-L-Cys mutant enzyme H343A Citrobacter freundii
4.1.99.2 1.2
-
 3-fluoro-L-Tyr mutant enzyme H343A Citrobacter freundii
4.1.99.2 2.2
-
 L-Tyr mutant enzyme H343A Citrobacter freundii
4.1.99.2 3.4
-
 S-methyl-L-Cys wild-type enzyme Citrobacter freundii
4.1.99.2 6.6
-
 S-ethyl-L-Cys wild-type enzyme Citrobacter freundii
4.1.99.2 11
-
 L-Ala wild-type enzyme Citrobacter freundii
4.1.99.2 11
-
 L-Ser mutant enzyme H343A Citrobacter freundii
4.1.99.2 12.99
-
 S-methyl-L-Cys mutant enzyme H343A Citrobacter freundii
4.1.99.2 18
-
 L-Ser wild-type enzyme Citrobacter freundii
4.1.99.2 32
-
 D-Ala wild-type enzyme, alpha,beta-elimination Citrobacter freundii
4.1.99.2 56.4
-
 S-methyl-L-Cys wild-type enzyme Citrobacter freundii
4.1.99.2 103
-
 L-Ala mutant enzyme H343A Citrobacter freundii

Organism

EC Number Organism UniProt Comment Textmining
4.1.99.2 Citrobacter freundii
-
 wild type enzyme and mutant enzyme H343A
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.1.99.2 mutant enzyme His343Ala Citrobacter freundii

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.1.99.2 1.9
-
-
 Citrobacter freundii

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.1.99.2 Ala + H2O L-Ala Citrobacter freundii ?
-
 ?
4.1.99.2 Ala + H2O D-Ala Citrobacter freundii ?
-
 ?
4.1.99.2 D-Ala + H2O
-
 Citrobacter freundii ?
-
 ?
4.1.99.2 S-(o-nitrophenyl)-L-Cys + H2O
-
 Citrobacter freundii ?
-
 ?
4.1.99.2 S-benzoyl-L-Cys + H2O
-
 Citrobacter freundii ?
-
 ?
4.1.99.2 S-benzyl-L-Cys + H2O
-
 Citrobacter freundii ? + pyruvate + NH3
-
 ?
4.1.99.2 S-ethyl-L-Cys + H2O
-
 Citrobacter freundii ?
-
 ?
4.1.99.2 S-methyl-L-Cys + H2O
-
 Citrobacter freundii ? + pyruvate + NH3
-
 ?
4.1.99.2 Ser + H2O L-Ser Citrobacter freundii ? + pyruvate + NH3
-
 ?
4.1.99.2 Tyr + H2O
-
 Citrobacter freundii phenol + pyruvate + NH3
-
 ?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.1.99.2 0.004
-
 D-Ala mutant enzyme H343A Citrobacter freundii
4.1.99.2 0.01
-
 D-Ala wild-type enzyme Citrobacter freundii
4.1.99.2 0.02
-
 S-benzoyl-L-Cys mutant enzyme H343A Citrobacter freundii
4.1.99.2 0.03
-
 L-Ala wild-type enzyme Citrobacter freundii
4.1.99.2 0.07
-
 L-Ser mutant enzyme H343A Citrobacter freundii
4.1.99.2 0.15
-
 S-methyl-L-Cys mutant enzyme H343A Citrobacter freundii
4.1.99.2 0.17
-
 L-Ser wild-type enzyme Citrobacter freundii
4.1.99.2 0.18
-
 3-fluoro-L-Tyr mutant enzyme H343A Citrobacter freundii
4.1.99.2 0.42
-
 L-Tyr mutant enzyme H343A Citrobacter freundii
4.1.99.2 0.52
-
 S-benzyl-L-Cys wild-type enzyme Citrobacter freundii
4.1.99.2 1.4
-
 3-fluoro-L-Tyr wild-type enzyme Citrobacter freundii
4.1.99.2 1.4
-
 S-ethyl-L-Cys mutant enzyme H343A Citrobacter freundii
4.1.99.2 1.55
-
 S-(o-nitrophenyl)-L-Cys mutant enzyme H343A Citrobacter freundii
4.1.99.2 3.5
-
 L-Tyr wild-type enzyme Citrobacter freundii
4.1.99.2 3.88
-
 S-ethyl-L-Cys wild-type enzyme Citrobacter freundii
4.1.99.2 5.1
-
 S-(o-nitrophenyl)-L-Cys wild-type enzyme Citrobacter freundii