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Literature summary for 3.1.1.11 extracted from

  • Fries, M.; Ihrig, J.; Brocklehurst, K.; Shevchik, V.E.; Pickersgill, R.W.
    Molecular basis of the activity of the phytopathogen pectin methylesterase (2007), EMBO J., 26, 3879-3887.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli strain NM522 Dickeya chrysanthemi

Crystallization (Commentary)

Crystallization (Comment) Organism
the inactive D178A PME mutant in complex with specifically methylated hexagalacturonates, 3 mg/ml protein, crystallization solutions contain 0.1 M MES, pH 6.5, 10% dioxane and 1.6 M ammonium sulfate, and dilutions of that crystallisation buffer with H2O, or 0.1 M MES, pH 6.5, and 12% w/v PEG 20000, X-ray diffraction structure determination and anaylsis at 1.7-1.9 A resolution Dickeya chrysanthemi

Protein Variants

Protein Variants Comment Organism
D178A site-directed mutagenesis, inactive mutant Dickeya chrysanthemi
D199A site-directed mutagenesis, inactive mutant Dickeya chrysanthemi
M306A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Dickeya chrysanthemi
Q153A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Dickeya chrysanthemi
Q177A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Dickeya chrysanthemi
R267A site-directed mutagenesis, inactive mutant Dickeya chrysanthemi
T272A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Dickeya chrysanthemi
V198A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Dickeya chrysanthemi
W269A site-directed mutagenesis, inactive mutant Dickeya chrysanthemi

Inhibitors

Inhibitors Comment Organism Structure
Polygalacturonate completely deesterified pectin, a competitive inhibitor of PME Dickeya chrysanthemi

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information steady-state kinetics, kinetic mechanism Dickeya chrysanthemi
0.06
-
Pectin mutant V198A, pH 7.0, 30°C Dickeya chrysanthemi
0.13
-
Pectin wild-type enzyme, pH 7.0, 30°C Dickeya chrysanthemi
0.22
-
Pectin mutant Q177A, pH 7.0, 30°C Dickeya chrysanthemi
0.53
-
Pectin mutant M306A, pH 7.0, 30°C Dickeya chrysanthemi
0.77
-
Pectin mutant T272A, pH 7.0, 30°C Dickeya chrysanthemi
2.2
-
Pectin mutant Q153A, pH 7.0, 30°C Dickeya chrysanthemi

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
pectin + H2O Dickeya chrysanthemi the enzyme catalyses the essential first step in bacterial invasion of plant tissues methanol + pectate
-
?

Organism

Organism UniProt Comment Textmining
Dickeya chrysanthemi
-
a plant pathogen, strain B374
-

Reaction

Reaction Comment Organism Reaction ID
pectin + n H2O = n methanol + pectate two conserved aspartates are the nucleophile and general acid-base in the reaction mechanism, respectively, the catalytic site is formed by the conserved resisues Asp178, Asp199, and Arg267, molecular basis of the processive action of the enzyme, overview Dickeya chrysanthemi

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information substrate specificity, overview Dickeya chrysanthemi ?
-
?
pectin + H2O the enzyme catalyses the essential first step in bacterial invasion of plant tissues Dickeya chrysanthemi methanol + pectate
-
?
pectin + H2O the enzyme shows a sequential pattern of demethylation due to the preferential binding of methylated sugar residues upstream of the catalytic site, and demethylated residues downstream, which drives the enzyme along the pectin molecule Dickeya chrysanthemi methanol + pectate
-
?

Subunits

Subunits Comment Organism
More the enzyme possesses a parallel beta-helix architecture, three dimensional structure of PME, overview Dickeya chrysanthemi

Synonyms

Synonyms Comment Organism
pectin methylesterase
-
Dickeya chrysanthemi
PME
-
Dickeya chrysanthemi

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Dickeya chrysanthemi

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
40
-
Pectin mutant Q177A, pH 7.0, 30°C Dickeya chrysanthemi
59
-
Pectin mutant M306A, pH 7.0, 30°C Dickeya chrysanthemi
68
-
Pectin mutant V198A, pH 7.0, 30°C Dickeya chrysanthemi
232
-
Pectin mutant Q153A, pH 7.0, 30°C Dickeya chrysanthemi
425
-
Pectin mutant T272A, pH 7.0, 30°C Dickeya chrysanthemi
450
-
Pectin wild-type enzyme, pH 7.0, 30°C Dickeya chrysanthemi

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
assay at Dickeya chrysanthemi

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
16
-
Polygalacturonate
-
Dickeya chrysanthemi