Literature summary for 1.1.1.274 extracted from

Sanli, G.; Banta, S.; Anderson, S.; Blaber, M.
Structural alteration of cofactor specificity in Corynebacterium 2,5-diketo-D-gluconic acid reductase (2004), Protein Sci., 13, 504-512.

Search for more literature for 1.1.1.274

Application

Application	Comment	Organism
biotechnology	enzyme is a target for the construction of a NADH-utilizing mutant strain in the industrial production of vitamin C	Corynebacterium sp.
synthesis	enzyme can be used in the industrial production of vitamin C	Corynebacterium sp.

Crystallization (Commentary)

Crystallization (Comment)	Organism
10 mg/ml purified recombinant F22Y/K232G/R238H/A272G mutant enzyme in complex with NADH in 25 mM Tris-HCl, pH 7.5, 1 mM NADH, hanging drop vapour diffusion method, room temperature, equal volumes, 0.005 ml each, of protein and crystallization solution against 1 ml reservoir crystallization solution containing 1.5 M lithium sulfate, 0.1 M Na-HEPES, pH 7.5, 7-10 days, X-ray diffraction structure determination and analysis at 2.0 A resolution, molecular replacment, molecular modeling of substrate and cofactor binding	Corynebacterium sp.

Crystallization (Comment)

Organism

10 mg/ml purified recombinant F22Y/K232G/R238H/A272G mutant enzyme in complex with NADH in 25 mM Tris-HCl, pH 7.5, 1 mM NADH, hanging drop vapour diffusion method, room temperature, equal volumes, 0.005 ml each, of protein and crystallization solution against 1 ml reservoir crystallization solution containing 1.5 M lithium sulfate, 0.1 M Na-HEPES, pH 7.5, 7-10 days, X-ray diffraction structure determination and analysis at 2.0 A resolution, molecular replacment, molecular modeling of substrate and cofactor binding

Corynebacterium sp.

Protein Variants

Protein Variants	Comment	Organism
A272G	mutation increases Km and kcat compared to the wild-type enzyme	Corynebacterium sp.
F22Y	mutation reduces Km and increases kcat by 50% compared to the wild-type enzyme	Corynebacterium sp.
F22Y/A272G	increased activity compared to the wild-type enzyme, substrate inhibition at substrate concentrations above 17.5 mM	Corynebacterium sp.
F22Y/K232G/R238H/A272G	mutant shows a higher activity with NADH compared to the wild-type enzyme	Corynebacterium sp.

General Stability

General Stability	Organism
isozyme A is more stable than isozyme B but less active	Corynebacterium sp.

Inhibitors

Inhibitors	Comment	Organism	Structure
2,5-didehydro-D-gluconate	substrate inhibition of isozyme B, not of isozyme A, at high concentrations	Corynebacterium sp.

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
29000	-	x * 29000	Corynebacterium sp.

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
2,5-didehydro-D-gluconate + NADPH	Corynebacterium sp.	step in the biosynthesis of L-ascorbic acid	2-oxo-L-gulonic acid + NADP+	-	?

Organism

Organism	UniProt	Comment	Textmining
Corynebacterium sp.	P06632	2 isozymes A and B	-

Reaction

Reaction	Comment	Organism	Reaction ID
2-dehydro-D-gluconate + NADP+ = 2,5-didehydro-D-gluconate + NADPH + H+	reaction mechanism of wild-type and F22Y/K232G/R238H/A272G mutant enzyme	Corynebacterium sp.	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
2,5-didehydro-D-gluconate + NADPH	step in the biosynthesis of L-ascorbic acid	Corynebacterium sp.	2-oxo-L-gulonic acid + NADP+	-	?
2,5-didehydro-D-gluconate + NADPH	i.e. 2,5-diketo-D-gluconic acid or 2,5-DKG, stereospecific reaction	Corynebacterium sp.	2-oxo-L-gulonic acid + NADP+	i.e. 2-keto-L-gulonic acid or 2-KLG, product is a precursor for L-ascorbic acid	?
additional information	isozyme A is more stable than isozyme B but less active	Corynebacterium sp.	?	-	?

Subunits

Subunits	Comment	Organism
?	x * 29000	Corynebacterium sp.

Synonyms

Synonyms	Comment	Organism
2,5-diketo-D-gluconic acid reductase	-	Corynebacterium sp.
2,5-DKGR	-	Corynebacterium sp.

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
additional information	-	thermodynamic stability study of wild-type and F22Y/K232G/R238H/A272G mutant enzyme	Corynebacterium sp.

Cofactor

Cofactor	Comment	Organism
additional information	cofactor binding structure of wild-type and F22Y/K232G/R238H/A272G mutant enzyme, involved Lys232, Ala272, and Arg238	Corynebacterium sp.
NADH	F22Y/K232G/R238H/A272G mutant enzyme	Corynebacterium sp.
NADPH	preferred cofactor of the wild-type enzyme	Corynebacterium sp.