Literature summary for 1.1.1.169 extracted from

Zheng, R.; Blanchard, J.S.
Identification of active site residues in E. coli ketopantoate reductase by mutagenesis and chemical rescue (2000), Biochemistry, 39, 16244-16251.

Search for more literature for 1.1.1.169

Activating Compound

Activating Compound	Comment	Organism	Structure
bromoethylamine	activation of enzyme from K176C mutant	Escherichia coli
ethylamine	activation of enzyme from K176A mutant	Escherichia coli
formate	E256A mutant activity greatly increased, twice as high at pH 7.2 as at pH 5.9	Escherichia coli
methylamine	activation of enzyme from K176A mutant	Escherichia coli
Propylamine	activation of enzyme from K176A mutant	Escherichia coli

Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli BL21(DE3)	Escherichia coli

Protein Variants

Protein Variants	Comment	Organism
D248A	site-directed mutagenesis, wild-type activity	Escherichia coli
E210A	site-directed mutagenesis, wild-type activity	Escherichia coli
E240A	site-directed mutagenesis, wild-type activity	Escherichia coli
E256A	site-directed mutagenesis, significant reduction in catalytic efficiency of enzyme	Escherichia coli
E256D	wild-type activity	Escherichia coli
K176A	site-directed mutagenesis, significant reduction in catalytic efficiency of enzyme	Escherichia coli
K176A/E256A	double mutant, no activity	Escherichia coli
K176C	wild-type activity	Escherichia coli
K72A	site-directed mutagenesis, wild-type activity	Escherichia coli
additional information	Lys176 acts as general acid in ketopantoate reduction and is involved in catalysis and ketopantoate binding, E256A functions in D-pantoate and ketopantoate binding in ketopantoate reductase	Escherichia coli

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.002	-	NADPH	+/-0.0003, mutant E256A	Escherichia coli
0.0029	-	NADPH	+/-0.0006, mutant E256D	Escherichia coli
0.0038	-	NADPH	+/-0.0003, mutant K176C, alkylated	Escherichia coli
0.004	-	NADPH	+/-0.0004	Escherichia coli
0.0066	-	NADPH	+/-0.0016, mutant K176C	Escherichia coli
0.016	-	NADPH	+/-0.003, mutant K176A	Escherichia coli
0.038	-	ketopantoate	+/-0.009, mutant K176C, alkylated	Escherichia coli
0.07	-	ketopantoate	+/-0.01, mutant K176C	Escherichia coli
0.12	-	ketopantoate	+/-0.008	Escherichia coli
0.95	-	ketopantoate	+/-0.29, mutant E256D	Escherichia coli
7.5	-	ketopantoate	+/-2.9, mutant K256A	Escherichia coli
40	-	ketopantoate	+/-6, mutant K176A	Escherichia coli

Metals/Ions

Metals/Ions	Comment	Organism	Structure
additional information	-	Escherichia coli

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
34000	-	-	Escherichia coli

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	-	expressed in BL21(DE3)	-

Purification (Commentary)

Purification (Comment)	Organism
homogeneity in a yield of 20-60 mg from 3-6 g of cells	Escherichia coli

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
additional information	-	no activity for double mutant K176A/E256A	Escherichia coli

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
alpha-ketopantoate + NADPH	-	Escherichia coli	D-pantoate + NADP+	-	r
additional information	no activity for double mutant K176A/E256A	Escherichia coli	?	-	?

Subunits

Subunits	Comment	Organism
monomer	-	Escherichia coli

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
additional information	-	activity decreases with pH increase, reduction of ketopantoic acid	Escherichia coli

Cofactor

Cofactor	Comment	Organism	Structure
NADP+	-	Escherichia coli
NADPH	-	Escherichia coli

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Release 2023.1 (January 2023)