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Literature summary extracted from

  • Qiu, X.; Tao, Y.; Zhu, Y.; Yuan, Y.; Zhang, Y.; Liu, H.; Gao, Y.; Teng, M.; Niu, L.
    Structural insights into decreased enzymatic activity induced by an insert sequence in mannonate dehydratase from Gram negative bacterium (2012), J. Struct. Biol., 180, 327-334.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.1.8 expressed in Escherichia coli as a His-tagged fusion protein Escherichia coli

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
4.2.1.8 crystal structures of ManD and its complex with D-mannonate is solved. It shows that the insert sequence forms two alpha helices locating above the active site. The two insert alpha helices introduce a loop that forms a cap covering the substrate binding pocket, which restricts the tunnels of substrate entering and product releasing from the active site Escherichia coli

Protein Variants

EC Number Protein Variants Comment Organism
4.2.1.8 E185D kcat decreased compared to wild-type, Km decreased compared to wild-type Escherichia coli
4.2.1.8 E185G kcat increased compared to wild-type, Km slightly decreased compared to wild-type Escherichia coli
4.2.1.8 E186D kcat decreased compared to wild-type, Km decreased compared to wild-type Escherichia coli
4.2.1.8 E186G kcat and Km increased compared to wild-type Escherichia coli
4.2.1.8 E63A/S64A inactive mutant Escherichia coli
4.2.1.8 H32G kcat and Km increased compared to wild-type Escherichia coli
4.2.1.8 N103A/D109A kcat decreased compared to wild-type, Km slightly decreased compared to wild-type Escherichia coli
4.2.1.8 N36G kcat increased compared to wild-type, Km slightly decreased compared to wild-type Escherichia coli
4.2.1.8 W110F inactive mutant Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.2.1.8 2.31
-
 D-mannonate mutant E185D, pH 7.5, 37°C Escherichia coli
4.2.1.8 3.6
-
 D-mannonate mutant E186D, pH 7.5, 37°C Escherichia coli
4.2.1.8 4.08
-
 D-mannonate mutant N103A/D109A, pH 7.5, 37°C Escherichia coli
4.2.1.8 4.55
-
 D-mannonate mutant E185G, pH 7.5, 37°C Escherichia coli
4.2.1.8 4.67
-
 D-mannonate mutant N36G, pH 7.5, 37°C Escherichia coli
4.2.1.8 4.79
-
 D-mannonate wild-type, pH 7.5, 37°C Escherichia coli
4.2.1.8 5.21
-
 D-mannonate mutant H32G, pH 7.5, 37°C Escherichia coli
4.2.1.8 5.3
-
 D-mannonate mutant E186G, pH 7.5, 37°C Escherichia coli

Organism

EC Number Organism UniProt Comment Textmining
4.2.1.8 Escherichia coli P24215
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.1.8 using Ni-NTA chromatography Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.1.8 D-Mannonate
-
 Escherichia coli 2-Dehydro-3-deoxy-D-gluconate + H2O
-
 ?

Subunits

EC Number Subunits Comment Organism
4.2.1.8 monomer crystal structure Escherichia coli

Synonyms

EC Number Synonyms Comment Organism
4.2.1.8 EcManD
-
 Escherichia coli
4.2.1.8 mannonate dehydratase
-
 Escherichia coli

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.2.1.8 37
-
 assay at Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.2.1.8 0.1
-
 D-mannonate mutant N103A/D109A, pH 7.5, 37°C Escherichia coli
4.2.1.8 0.23
-
 D-mannonate mutant E185D, pH 7.5, 37°C Escherichia coli
4.2.1.8 0.29
-
 D-mannonate mutant E186D, pH 7.5, 37°C Escherichia coli
4.2.1.8 0.81
-
 D-mannonate wild-type, pH 7.5, 37°C Escherichia coli
4.2.1.8 0.94
-
 D-mannonate mutant E185G, pH 7.5, 37°C Escherichia coli
4.2.1.8 1.1
-
 D-mannonate mutant E186G, pH 7.5, 37°C Escherichia coli
4.2.1.8 1.29
-
 D-mannonate mutant H32G, pH 7.5, 37°C Escherichia coli
4.2.1.8 1.4
-
 D-mannonate mutant N36G, pH 7.5, 37°C Escherichia coli

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.2.1.8 7.5
-
 assay at Escherichia coli