Literature summary for 4.2.1.8 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.

Search for more literature for 4.2.1.8

Cloned(Commentary)

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

Crystallization (Commentary)

Crystallization (Comment)	Organism
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

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

KM Value [mM]

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

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	P24215	-	-

Purification (Commentary)

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

Substrates and Products (Substrate)

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

Subunits

Subunits	Comment	Organism
monomer	crystal structure	Escherichia coli

Synonyms

Synonyms	Comment	Organism
EcManD	-	Escherichia coli
mannonate dehydratase	-	Escherichia coli

Temperature Optimum [°C]

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

Turnover Number [1/s]

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

pH Optimum

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

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Release 2023.1 (January 2023)