Literature summary for 1.3.1.20 extracted from

Carbone, V.; Endo, S.; Sumii, R.; Chung, R.; Matsunaga, T.; Hara, A.; El-Kabbani, O.
Structures of dimeric dihydrodiol dehydrogenase apoenzyme and inhibitor complex: Probing the subunit interface with site-directed mutagenesis (2008), Proteins Struct. Funct. Genet., 70, 176-187.

Cloned(Commentary)

Cloned (Comment)	Organism
in Escherichia coli BL21(DE3)	Macaca fascicularis

Crystallization (Commentary)

Crystallization (Comment)	Organism
dimeric apoenzyme (at 2.0 A resolution) and enzyme-inhibitor complex (at 2.9 A resolution), crystals are grown using droplets consisting of the dihydrodiol dehydrogenase/NADPH solution mixed with a matching volume containing 1.5 M ammonium phosphate and 0.1 M sodium citrate at pH 5.6, tertiary structure is formed by a classical dinucleotide binding fold comprising of 2 beta-alpha-beta-alpha-beta motifs at the N-terminus and an eight-stranded, predominantly antiparallel beta-sheet at the C-terminus	Macaca fascicularis

Crystallization (Comment)

Organism

dimeric apoenzyme (at 2.0 A resolution) and enzyme-inhibitor complex (at 2.9 A resolution), crystals are grown using droplets consisting of the dihydrodiol dehydrogenase/NADPH solution mixed with a matching volume containing 1.5 M ammonium phosphate and 0.1 M sodium citrate at pH 5.6, tertiary structure is formed by a classical dinucleotide binding fold comprising of 2 beta-alpha-beta-alpha-beta motifs at the N-terminus and an eight-stranded, predominantly antiparallel beta-sheet at the C-terminus

Macaca fascicularis

Protein Variants

Protein Variants	Comment	Organism
R148A	mutagenesis is performed using a QuickChange site-directed mutagenesis kit, dehydrogenase activity is comparable to that of the wild type	Macaca fascicularis
R148A/R202A	mutagenesis is performed using a QuickChange site-directedmutagenesis kit	Macaca fascicularis
R202A	mutagenesis is performed using a QuickChange site-directed mutagenesis kit, dehydrogenase activity is comparable to that of the wild type	Macaca fascicularis

Inhibitors

Inhibitors	Comment	Organism	Structure
4-hydroxyacetophenone	-	Macaca fascicularis

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
70000	-	determined by gel filtration	Macaca fascicularis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
additional information	Macaca fascicularis	dimeric dihydrodiol dehydrogenase catalyses the nicotinamide adenine dinucleotide phosphate NADP+-dependent oxidation of trans-dihydrodiols of aromatic hydrocarbons to their corresponding catechols, dimeric dihydrodiol dehydrogenase is also competent in the NADPH-driven reduction of a limited number of aldehyde and dicarbonyl compounds such as 3-deoxyglucosone, camphorquinone, and 3-nitrobenzaldehyde	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Macaca fascicularis	Q9TQS6	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant enzymes are purified from the 12000g supernatants of the homogenates of the cells, the single mutant enzymes are partially purified by the ammonium sulphate fractionation and chromatography on a Sephadex G-100 column, the wild type and double mutant enzyme are further purified by chromatography on a Red Sepharose column	Macaca fascicularis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.04	-	R148A and R202A mutant, 25 mM Tris-HCl, pH 7.5, 0.25 mM NADP+	Macaca fascicularis
0.05	-	wild type enzyme in dimeric form, 25 mM Tris-HCl, pH 7.5, 0.25 mM NADP+	Macaca fascicularis
1.2	-	monomeric form	Macaca fascicularis

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
additional information	dimeric dihydrodiol dehydrogenase catalyses the nicotinamide adenine dinucleotide phosphate NADP+-dependent oxidation of trans-dihydrodiols of aromatic hydrocarbons to their corresponding catechols, dimeric dihydrodiol dehydrogenase is also competent in the NADPH-driven reduction of a limited number of aldehyde and dicarbonyl compounds such as 3-deoxyglucosone, camphorquinone, and 3-nitrobenzaldehyde	Macaca fascicularis	?	-	?

Subunits

Subunits	Comment	Organism
dimer	site-directed mutagenesis has demonstrated that the intact dimer is not essential for catalytic activity	Macaca fascicularis
More	the dissociation of DD into the monomer largely affects the stability rather than catalytic activity	Macaca fascicularis

Synonyms

Synonyms	Comment	Organism
dihydrodiol dehydrogenase	-	Macaca fascicularis

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
37	-	no activity of the 37°C-inactivated R148A/R202A mutant is recovered by the addition of 0.1 mM NADP+ and 40% glycerol	Macaca fascicularis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	assay at	Macaca fascicularis

Cofactor

Cofactor	Comment	Organism	Structure
NADP+	-	Macaca fascicularis
NADPH	-	Macaca fascicularis

IC50 Value

IC50 Value	IC50 Value Maximum	Comment	Organism	Inhibitor	Structure
0.00097	-	the hydroxyl group of 4-hydroxyacetophenone forms hydrogen bond interactions with the side-chain oxygen and the main chain nitrogen of Asp280, the benzene ring and methyl group make a number of hydrophobic contacts with Phe279, Leu158 and Leu177	Macaca fascicularis	4-hydroxyacetophenone