Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 4.2.1.51 extracted from

  • Hsu, S.K.; Lin, L.L.; Lo, H.H.; Hsu, W.H.
    Mutational analysis of feedback inhibition and catalytic sites of prephenate dehydratase from Corynebacterium glutamicum (2004), Arch. Microbiol., 181, 237-244.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of the His-tagged wild-type and mutant enzymes in Escherichia coli JM109 Corynebacterium glutamicum

Protein Variants

Protein Variants Comment Organism
E64D constructed by PCR-based random mutagenesis and complementation analysis, 15% reduced activity compared to the wild-type enzyme, 4.5% increased Km, 1.7fold increased kcat Corynebacterium glutamicum
E64Q constructed by PCR-based random mutagenesis and complementation analysis, inactive mutant Corynebacterium glutamicum
E64S constructed by PCR-based random mutagenesis and complementation analysis, inactive mutant Corynebacterium glutamicum
E64V constructed by PCR-based random mutagenesis and complementation analysis, inactive mutant Corynebacterium glutamicum
F185L constructed by PCR-based random mutagenesis and complementation analysis, inactive mutant Corynebacterium glutamicum
F185Y constructed by PCR-based random mutagenesis and complementation analysis, 96% reduced activity compared to the wild-type enzyme, 26% increased Km Corynebacterium glutamicum
additional information alterations of Asp76, Glu89, His115, and Arg236 do not cause significant changes in the kinetics properties Corynebacterium glutamicum
R184L constructed by PCR-based random mutagenesis and complementation analysis, 50% reduced activity compared to the wild-type enzyme Corynebacterium glutamicum
S99A constructed by PCR-based random mutagenesis and complementation analysis, insensitive to inhibition by phenylalanine Corynebacterium glutamicum
S99C constructed by PCR-based random mutagenesis and complementation analysis, insensitive to inhibition by phenylalanine Corynebacterium glutamicum
S99L constructed by PCR-based random mutagenesis and complementation analysis, insensitive to inhibition by phenylalanine Corynebacterium glutamicum
S99M constructed by PCR-based random mutagenesis and complementation analysis, 30% reduced activity compared to the wild-type enzyme, insensitive to inhibition by phenylalanine Corynebacterium glutamicum
T183A constructed by PCR-based random mutagenesis and complementation analysis, inactive mutant Corynebacterium glutamicum
T183S constructed by PCR-based random mutagenesis and complementation analysis, highly reduced activity Corynebacterium glutamicum
T183Y constructed by PCR-based random mutagenesis and complementation analysis, inactive mutant Corynebacterium glutamicum

Inhibitors

Inhibitors Comment Organism Structure
L-phenylalanine feedback inhibition, Ser99 is involved, mutants S99M, S99T, S99A, S99C, or S99L are not sensitive to inhibition Corynebacterium glutamicum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.07
-
prephenate wild-type enzyme, pH 7.5, 30°C Corynebacterium glutamicum
0.08
-
prephenate mutant S99T, pH 7.5, 30°C Corynebacterium glutamicum
0.09
-
prephenate mutant S99C, pH 7.5, 30°C Corynebacterium glutamicum
0.13
-
prephenate mutant S99A, pH 7.5, 30°C Corynebacterium glutamicum
0.18
-
prephenate mutant S99M, pH 7.5, 30°C Corynebacterium glutamicum
0.29
-
prephenate mutant E164D, pH 7.5, 30°C Corynebacterium glutamicum
0.46
-
prephenate mutant T183S, pH 7.5, 30°C Corynebacterium glutamicum
0.61
-
prephenate mutant R184L, pH 7.5, 30°C Corynebacterium glutamicum
1.73
-
prephenate mutant F185Y, pH 7.5, 30°C Corynebacterium glutamicum

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
39000
-
x * 39000, recombinant wild-type and mutant enzymes, SDS-PAGE Corynebacterium glutamicum

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
prephenate Corynebacterium glutamicum enzyme is a key regulatory enzyme in the phenylalanine-specific pathway phenylpyruvate + H2O + CO2
-
?
prephenate Corynebacterium glutamicum CCRC 11384 enzyme is a key regulatory enzyme in the phenylalanine-specific pathway phenylpyruvate + H2O + CO2
-
?

Organism

Organism UniProt Comment Textmining
Corynebacterium glutamicum
-
-
-
Corynebacterium glutamicum CCRC 11384
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli Corynebacterium glutamicum

Reaction

Reaction Comment Organism Reaction ID
prephenate = phenylpyruvate + H2O + CO2 this enzyme in the enteric bacteria also possesses chorismate mutase activity, EC 5.4.99.5, and converts chorismate into prephenate, residues Glu64, Thr183, Arg184, and Phe185 might be involved in catalysis and substrate binding Corynebacterium glutamicum

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.32
-
mutant F185Y, pH 7.5, 30°C Corynebacterium glutamicum
0.39
-
mutant T183S, pH 7.5, 30°C Corynebacterium glutamicum
2.37
-
mutant S99A, pH 7.5, 30°C Corynebacterium glutamicum
4.31
-
mutant R184L, pH 7.5, 30°C Corynebacterium glutamicum
6.66
-
mutant S99M, pH 7.5, 30°C Corynebacterium glutamicum
6.88
-
mutant S99C, pH 7.5, 30°C Corynebacterium glutamicum
7.59
-
mutant S99T, pH 7.5, 30°C Corynebacterium glutamicum
8.1
-
mutant E164D, pH 7.5, 30°C Corynebacterium glutamicum
9.48
-
wild-type enzyme, pH 7.5, 30°C Corynebacterium glutamicum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
prephenate
-
Corynebacterium glutamicum phenylpyruvate + H2O + CO2
-
?
prephenate enzyme is a key regulatory enzyme in the phenylalanine-specific pathway Corynebacterium glutamicum phenylpyruvate + H2O + CO2
-
?
prephenate
-
Corynebacterium glutamicum CCRC 11384 phenylpyruvate + H2O + CO2
-
?
prephenate enzyme is a key regulatory enzyme in the phenylalanine-specific pathway Corynebacterium glutamicum CCRC 11384 phenylpyruvate + H2O + CO2
-
?

Subunits

Subunits Comment Organism
? x * 39000, recombinant wild-type and mutant enzymes, SDS-PAGE Corynebacterium glutamicum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Corynebacterium glutamicum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.01
-
prephenate mutant F185Y, pH 7.5, 30°C Corynebacterium glutamicum
0.01
-
prephenate mutant T183S, pH 7.5, 30°C Corynebacterium glutamicum
0.03
-
prephenate mutant S99A, pH 7.5, 30°C Corynebacterium glutamicum
0.09
-
prephenate mutant S99C, pH 7.5, 30°C Corynebacterium glutamicum
0.11
-
prephenate mutant S99T, pH 7.5, 30°C Corynebacterium glutamicum
0.12
-
prephenate mutant S99M, pH 7.5, 30°C Corynebacterium glutamicum
0.12
-
prephenate wild-type enzyme, pH 7.5, 30°C Corynebacterium glutamicum
0.13
-
prephenate mutant R184L, pH 7.5, 30°C Corynebacterium glutamicum
0.21
-
prephenate mutant E164D, pH 7.5, 30°C Corynebacterium glutamicum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Corynebacterium glutamicum