Literature summary for 1.1.1.82 extracted from

Kawakami, R.; Sakuraba, H.; Goda, S.; Tsuge, H.; Ohshima, T.
Refolding, characterization and crystal structure of (S)-malate dehydrogenase from the hyperthermophilic archaeon Aeropyrum pernix (2009), Biochim. Biophys. Acta, 1794, 1496-1504.

Cloned(Commentary)

Cloned (Comment)	Organism
expression of MDH in Escherichia coli strain BL21(DE3) in inclusion bodies	Aeropyrum pernix

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified refolded reconbinant enzyme, sitting drop vapor diffusion method, 0.002 ml of protein solution containing 8.9 mg/ml protein in 50 mM Tris-HCl, pH7.5, containing 0.2 M NaCl, are mixed with an equal volume of the reservoir solution, comprised of 0.1 M CHES, pH 9.5, and 40% PEG 600, and equilibrated against 0.15 ml of reservoir solution at 20°C, 2 weeks, X-ray diffraction structure determination and analysis at 2.9 A resolution and room temperature	Aeropyrum pernix

Crystallization (Comment)

Organism

purified refolded reconbinant enzyme, sitting drop vapor diffusion method, 0.002 ml of protein solution containing 8.9 mg/ml protein in 50 mM Tris-HCl, pH7.5, containing 0.2 M NaCl, are mixed with an equal volume of the reservoir solution, comprised of 0.1 M CHES, pH 9.5, and 40% PEG 600, and equilibrated against 0.15 ml of reservoir solution at 20°C, 2 weeks, X-ray diffraction structure determination and analysis at 2.9 A resolution and room temperature

Aeropyrum pernix

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.019	-	(S)-malate	pH 10.0, 50°C, with cofactor NADP+, recombinant enzyme	Aeropyrum pernix
0.12	-	(S)-malate	pH 10.0, 50°C, with cofactor NAD+, recombinant enzyme	Aeropyrum pernix
0.2	-	(2S,3R)-tartrate	pH 10.0, 50°C, with cofactor NAD+, recombinant enzyme	Aeropyrum pernix
1.2	-	(2S,3S)-tartrate	pH 10.0, 50°C, with cofactor NAD+, recombinant enzyme	Aeropyrum pernix
2	3	(2S,3S)-tartrate	pH 10.0, 50°C, with cofactor NADP+, recombinant enzyme	Aeropyrum pernix
5.8	-	(2S,3R)-tartrate	pH 10.0, 50°C, with cofactor NADP+, recombinant enzyme	Aeropyrum pernix

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
33489	-	4 * 34000, recombinant enzyme, SDS-PAGE, 4 * 33489, sequence calculation	Aeropyrum pernix
34000	-	4 * 34000, recombinant enzyme, SDS-PAGE, 4 * 33489, sequence calculation	Aeropyrum pernix
110000	-	recombinant enzyme, gel filtration	Aeropyrum pernix

Organism

Organism	UniProt	Comment	Textmining
Aeropyrum pernix	Q9YEA1	a strict aerobic hyperthermophilic archaeon isolated from a coastal thermal vent in Japan	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant MDH from Escherichia coli inclusion bodies by solubilization, refolding, dialysis, and gel filtration	Aeropyrum pernix

Renatured (Commentary)

Renatured (Comment)	Organism
solubilization and refolding of recombinant MDH from Escherichia coli inclusion bodies, the enzyme is dissolved in 6 M guanidine-HCl and gradually refolded to the active enzyme through dilution of the denaturant, conditions: refolding buffer 1.5 L of 0.1 M Tris-HCl, pH 7.5, containing 2 mM EDTA and 0.4 M L-arginine, incubation for 36 h at 4°C	Aeropyrum pernix

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
(2S,3R)-tartrate + NAD(P)+	-	Aeropyrum pernix	? + NAD(P)H + H+	-	?
(2S,3S)-tartrate + NAD(P)+	-	Aeropyrum pernix	? + NAD(P)H + H+	-	?
(S)-malate + NAD(P)+	-	Aeropyrum pernix	oxaloacetate + NAD(P)H + H+	-	?

Subunits

Subunits	Comment	Organism
More	structure qand sequence comaprsion, overall structure, overview	Aeropyrum pernix
tetramer	4 * 34000, recombinant enzyme, SDS-PAGE, 4 * 33489, sequence calculation	Aeropyrum pernix

Synonyms

Synonyms	Comment	Organism
(S)-malate dehydrogenase	-	Aeropyrum pernix
MDH	-	Aeropyrum pernix

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
95	-	above	Aeropyrum pernix

Temperature Range [°C]

Temperature Minimum [°C]	Temperature Maximum [°C]	Comment	Organism
50	95	-	Aeropyrum pernix

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
50	-	purified recombinant enzyme, 30 min, pH 5.0-10.5, stable	Aeropyrum pernix
90	-	purified recombinant enzyme, 10 min, stable. The hyperthermostability of the Aeropyrum pernix MDH is likely attributable to its smaller cavity volume and larger numbers of ion pairs and ion-pair networks, but the molecular strategy for thermostability may be specific for each enzyme	Aeropyrum pernix
100	-	purified recombinant enzyme, 10 min, stable. The hyperthermostability of the Aeropyrum pernix MDH is likely attributable to its smaller cavity volume and larger numbers of ion pairs and ion-pair networks, but the molecular strategy for thermostability may be specific for each enzyme	Aeropyrum pernix

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
0.16	-	(2S,3R)-tartrate	pH 10.0, 50°C, with cofactor NAD+, recombinant enzyme	Aeropyrum pernix
0.2	-	(2S,3R)-tartrate	pH 10.0, 50°C, with cofactor NADP+, recombinant enzyme	Aeropyrum pernix
0.37	-	(2S,3S)-tartrate	pH 10.0, 50°C, with cofactor NAD+, recombinant enzyme	Aeropyrum pernix
1.4	-	(S)-malate	pH 10.0, 50°C, with cofactor NADP+, recombinant enzyme	Aeropyrum pernix
2.6	-	(S)-malate	pH 10.0, 50°C, with cofactor NAD+, recombinant enzyme	Aeropyrum pernix
4.1	-	(2S,3S)-tartrate	pH 10.0, 50°C, with cofactor NADP+, recombinant enzyme	Aeropyrum pernix

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
11	-	-	Aeropyrum pernix

pH Stability

pH Stability	pH Stability Maximum	Comment	Organism
5	10.5	purified recombinant enzyme, 30 min, 50°C, stable	Aeropyrum pernix

Cofactor

Cofactor	Comment	Organism	Structure
NAD+	NADP+ is the preferred cofactor compared to NAD+ with substrate (S)-malate, while NAD+ is preferred with substrate tartrate. Ala at position 53 is responsible for coenzyme specificity, and the next residue, Arg, is important for NADP+ binding, structural analysis and comparison	Aeropyrum pernix
NADP+	NADP+ is the preferred cofactor compared to NAD+ with substrate (S)-malate, while NAD+ is preferred with substrate tartrate. Ala at position 53 is responsible for coenzyme specificity, and the next residue, Arg, is important for NADP+ binding, structural analysis and comparison	Aeropyrum pernix