BRENDA - Enzyme Database

Characterization of two ?-decarboxylating dehydrogenases from Sulfolobus acidocaldarius

Takahashi, K.; Nakanishi, F.; Tomita, T.; Akiyama, N.; Lassak, K.; Albers, S.V.; Kuzuyama, T.; Nishiyama, M.; Extremophiles 20, 843-853 (2016)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
1.1.1.85
expressed in Escherichia coli BL21(DE3)Codon-Plus-RIL cells
Sulfolobus acidocaldarius
1.1.1.286
gene saci_2375, DNA and amino acid sequence determination and analysis, recombinant expression in Escherichia coli strain BL21(DE3)Codon-Plus-RIL
Sulfolobus acidocaldarius
Crystallization (Commentary)
EC Number
Crystallization
Organism
1.1.1.85
in complex with (2R,3S)-3-isopropylmalate and Mg2+, hanging drop vapor diffusion method, using 2.0 M (NH4)2SO4, 200 mM NaCl, and 100 mM sodium cacodylate pH 6.5
Sulfolobus acidocaldarius
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.85
0.005
-
(2R,3S)-3-isopropylmalate
at pH 8.0 and 60°C
Sulfolobus acidocaldarius
1.1.1.85
0.12
-
NAD+
at pH 8.0 and 60°C
Sulfolobus acidocaldarius
1.1.1.85
5.9
-
homoisocitrate
at pH 8.0 and 60°C
Sulfolobus acidocaldarius
1.1.1.286
0.011
-
homoisocitrate
pH 8.0, 60°C, recombinant enzyme
Sulfolobus acidocaldarius
1.1.1.286
0.027
-
isocitrate
pH 8.0, 60°C, recombinant enzyme
Sulfolobus acidocaldarius
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
1.1.1.85
Mg2+
required
Sulfolobus acidocaldarius
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.1.85
36500
-
4 * 36500, SDS-PAGE
Sulfolobus acidocaldarius
1.1.1.85
130000
-
gel filtration
Sulfolobus acidocaldarius
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.1.1.85
(2R,3S)-3-isopropylmalate + NAD+
Sulfolobus acidocaldarius
-
(2S)-2-isopropyl-3-oxosuccinate + NADH + H+
-
-
?
1.1.1.85
(2R,3S)-3-isopropylmalate + NAD+
Sulfolobus acidocaldarius MW001
-
(2S)-2-isopropyl-3-oxosuccinate + NADH + H+
-
-
?
1.1.1.286
homoisocitrate + NADP+
Sulfolobus acidocaldarius
-
2-oxoadipate + CO2 + NADPH + H+
-
-
r
1.1.1.286
homoisocitrate + NADP+
Sulfolobus acidocaldarius MW001
-
2-oxoadipate + CO2 + NADPH + H+
-
-
r
1.1.1.286
isocitrate + NADP+
Sulfolobus acidocaldarius
-
2-oxoglutarate + CO2 + NADPH
-
-
r
1.1.1.286
isocitrate + NADP+
Sulfolobus acidocaldarius MW001
-
2-oxoglutarate + CO2 + NADPH
-
-
r
1.1.1.286
additional information
Sulfolobus acidocaldarius
Saci_2375 exhibits distinct and similar activities for isocitrate and homoisocitrate, but no detectable activity for 3-isopropylmalate. Saci_2375 is a dual function enzyme serving as isocitrate-homoisocitrate dehydrogenase
?
-
-
-
1.1.1.286
additional information
Sulfolobus acidocaldarius MW001
Saci_2375 exhibits distinct and similar activities for isocitrate and homoisocitrate, but no detectable activity for 3-isopropylmalate. Saci_2375 is a dual function enzyme serving as isocitrate-homoisocitrate dehydrogenase
?
-
-
-
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.1.1.85
Sulfolobus acidocaldarius
Q4JB37
-
-
1.1.1.85
Sulfolobus acidocaldarius MW001
Q4JB37
-
-
1.1.1.286
Sulfolobus acidocaldarius
Q4J6C9
-
-
1.1.1.286
Sulfolobus acidocaldarius MW001
Q4J6C9
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.1.1.85
DE52 column chromatography, phenyl Sepharose column chromatography, and Superdex 200 gel filtration
Sulfolobus acidocaldarius
1.1.1.286
recombinant enzyme from Escherichia coli strain BL21(DE3)Codon-Plus-RIL b anion exchange chromatography, ammonium sulfate fractionation, hydropobic interaction chromatography, ultrafiltration, and gel filtration
Sulfolobus acidocaldarius
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.85
(2R,3S)-3-isopropylmalate + NAD+
-
738195
Sulfolobus acidocaldarius
(2S)-2-isopropyl-3-oxosuccinate + NADH + H+
-
-
-
?
1.1.1.85
(2R,3S)-3-isopropylmalate + NAD+
-
738195
Sulfolobus acidocaldarius MW001
(2S)-2-isopropyl-3-oxosuccinate + NADH + H+
-
-
-
?
1.1.1.85
homoisocitrate + NAD+
-
738195
Sulfolobus acidocaldarius
? + NADH + H+
-
-
-
?
1.1.1.85
homoisocitrate + NAD+
-
738195
Sulfolobus acidocaldarius MW001
? + NADH + H+
-
-
-
?
1.1.1.85
additional information
the enzyme does not show detectable activity for isocitrate using NAD+ as a coenzyme, and is inactive with NADP+
738195
Sulfolobus acidocaldarius
?
-
-
-
-
1.1.1.85
additional information
the enzyme does not show detectable activity for isocitrate using NAD+ as a coenzyme, and is inactive with NADP+
738195
Sulfolobus acidocaldarius MW001
?
-
-
-
-
1.1.1.286
homoisocitrate + NADP+
-
738195
Sulfolobus acidocaldarius
2-oxoadipate + CO2 + NADPH + H+
-
-
-
r
1.1.1.286
homoisocitrate + NADP+
-
738195
Sulfolobus acidocaldarius MW001
2-oxoadipate + CO2 + NADPH + H+
-
-
-
r
1.1.1.286
isocitrate + NADP+
-
738195
Sulfolobus acidocaldarius
2-oxoglutarate + CO2 + NADPH
-
-
-
r
1.1.1.286
isocitrate + NADP+
-
738195
Sulfolobus acidocaldarius MW001
2-oxoglutarate + CO2 + NADPH
-
-
-
r
1.1.1.286
additional information
Saci_2375 exhibits distinct and similar activities for isocitrate and homoisocitrate, but no detectable activity for 3-isopropylmalate. Saci_2375 is a dual function enzyme serving as isocitrate-homoisocitrate dehydrogenase
738195
Sulfolobus acidocaldarius
?
-
-
-
-
1.1.1.286
additional information
Saci_2375 exhibits distinct and similar activities for isocitrate and homoisocitrate, but no detectable activity for 3-isopropylmalate. Saci_2375 is a dual function enzyme serving as isocitrate-homoisocitrate dehydrogenase
738195
Sulfolobus acidocaldarius MW001
?
-
-
-
-
Subunits
EC Number
Subunits
Commentary
Organism
1.1.1.85
homotetramer
4 * 36500, SDS-PAGE
Sulfolobus acidocaldarius
1.1.1.286
tetramer
dimer of dimers
Sulfolobus acidocaldarius
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.1.1.286
60
-
assay at
Sulfolobus acidocaldarius
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.1.1.85
0.26
-
homoisocitrate
at pH 8.0 and 60°C
Sulfolobus acidocaldarius
1.1.1.85
8.5
-
(2R,3S)-3-isopropylmalate
at pH 8.0 and 60°C
Sulfolobus acidocaldarius
1.1.1.286
37
-
homoisocitrate
pH 8.0, 60°C, recombinant enzyme
Sulfolobus acidocaldarius
1.1.1.286
61
-
isocitrate
pH 8.0, 60°C, recombinant enzyme
Sulfolobus acidocaldarius
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.286
8
-
assay at
Sulfolobus acidocaldarius
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.85
NAD+
dependent on
Sulfolobus acidocaldarius
1.1.1.286
NADP+
Saci_2375 is an NADP+-dependent enzyme with the ability to function as ICDH, EC 1.1.1.42, and HICDH, EC 1.1.1.87
Sulfolobus acidocaldarius
1.1.1.286
NADPH
Saci_2375 is an NADP+-dependent enzyme with the ability to function as ICDH, EC 1.1.1.42, and HICDH, EC 1.1.1.87
Sulfolobus acidocaldarius
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.85
expressed in Escherichia coli BL21(DE3)Codon-Plus-RIL cells
Sulfolobus acidocaldarius
1.1.1.286
gene saci_2375, DNA and amino acid sequence determination and analysis, recombinant expression in Escherichia coli strain BL21(DE3)Codon-Plus-RIL
Sulfolobus acidocaldarius
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.85
NAD+
dependent on
Sulfolobus acidocaldarius
1.1.1.286
NADP+
Saci_2375 is an NADP+-dependent enzyme with the ability to function as ICDH, EC 1.1.1.42, and HICDH, EC 1.1.1.87
Sulfolobus acidocaldarius
1.1.1.286
NADPH
Saci_2375 is an NADP+-dependent enzyme with the ability to function as ICDH, EC 1.1.1.42, and HICDH, EC 1.1.1.87
Sulfolobus acidocaldarius
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
1.1.1.85
in complex with (2R,3S)-3-isopropylmalate and Mg2+, hanging drop vapor diffusion method, using 2.0 M (NH4)2SO4, 200 mM NaCl, and 100 mM sodium cacodylate pH 6.5
Sulfolobus acidocaldarius
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.85
0.005
-
(2R,3S)-3-isopropylmalate
at pH 8.0 and 60°C
Sulfolobus acidocaldarius
1.1.1.85
0.12
-
NAD+
at pH 8.0 and 60°C
Sulfolobus acidocaldarius
1.1.1.85
5.9
-
homoisocitrate
at pH 8.0 and 60°C
Sulfolobus acidocaldarius
1.1.1.286
0.011
-
homoisocitrate
pH 8.0, 60°C, recombinant enzyme
Sulfolobus acidocaldarius
1.1.1.286
0.027
-
isocitrate
pH 8.0, 60°C, recombinant enzyme
Sulfolobus acidocaldarius
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
1.1.1.85
Mg2+
required
Sulfolobus acidocaldarius
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.1.85
36500
-
4 * 36500, SDS-PAGE
Sulfolobus acidocaldarius
1.1.1.85
130000
-
gel filtration
Sulfolobus acidocaldarius
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.1.1.85
(2R,3S)-3-isopropylmalate + NAD+
Sulfolobus acidocaldarius
-
(2S)-2-isopropyl-3-oxosuccinate + NADH + H+
-
-
?
1.1.1.85
(2R,3S)-3-isopropylmalate + NAD+
Sulfolobus acidocaldarius MW001
-
(2S)-2-isopropyl-3-oxosuccinate + NADH + H+
-
-
?
1.1.1.286
homoisocitrate + NADP+
Sulfolobus acidocaldarius
-
2-oxoadipate + CO2 + NADPH + H+
-
-
r
1.1.1.286
homoisocitrate + NADP+
Sulfolobus acidocaldarius MW001
-
2-oxoadipate + CO2 + NADPH + H+
-
-
r
1.1.1.286
isocitrate + NADP+
Sulfolobus acidocaldarius
-
2-oxoglutarate + CO2 + NADPH
-
-
r
1.1.1.286
isocitrate + NADP+
Sulfolobus acidocaldarius MW001
-
2-oxoglutarate + CO2 + NADPH
-
-
r
1.1.1.286
additional information
Sulfolobus acidocaldarius
Saci_2375 exhibits distinct and similar activities for isocitrate and homoisocitrate, but no detectable activity for 3-isopropylmalate. Saci_2375 is a dual function enzyme serving as isocitrate-homoisocitrate dehydrogenase
?
-
-
-
1.1.1.286
additional information
Sulfolobus acidocaldarius MW001
Saci_2375 exhibits distinct and similar activities for isocitrate and homoisocitrate, but no detectable activity for 3-isopropylmalate. Saci_2375 is a dual function enzyme serving as isocitrate-homoisocitrate dehydrogenase
?
-
-
-
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.85
DE52 column chromatography, phenyl Sepharose column chromatography, and Superdex 200 gel filtration
Sulfolobus acidocaldarius
1.1.1.286
recombinant enzyme from Escherichia coli strain BL21(DE3)Codon-Plus-RIL b anion exchange chromatography, ammonium sulfate fractionation, hydropobic interaction chromatography, ultrafiltration, and gel filtration
Sulfolobus acidocaldarius
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.85
(2R,3S)-3-isopropylmalate + NAD+
-
738195
Sulfolobus acidocaldarius
(2S)-2-isopropyl-3-oxosuccinate + NADH + H+
-
-
-
?
1.1.1.85
(2R,3S)-3-isopropylmalate + NAD+
-
738195
Sulfolobus acidocaldarius MW001
(2S)-2-isopropyl-3-oxosuccinate + NADH + H+
-
-
-
?
1.1.1.85
homoisocitrate + NAD+
-
738195
Sulfolobus acidocaldarius
? + NADH + H+
-
-
-
?
1.1.1.85
homoisocitrate + NAD+
-
738195
Sulfolobus acidocaldarius MW001
? + NADH + H+
-
-
-
?
1.1.1.85
additional information
the enzyme does not show detectable activity for isocitrate using NAD+ as a coenzyme, and is inactive with NADP+
738195
Sulfolobus acidocaldarius
?
-
-
-
-
1.1.1.85
additional information
the enzyme does not show detectable activity for isocitrate using NAD+ as a coenzyme, and is inactive with NADP+
738195
Sulfolobus acidocaldarius MW001
?
-
-
-
-
1.1.1.286
homoisocitrate + NADP+
-
738195
Sulfolobus acidocaldarius
2-oxoadipate + CO2 + NADPH + H+
-
-
-
r
1.1.1.286
homoisocitrate + NADP+
-
738195
Sulfolobus acidocaldarius MW001
2-oxoadipate + CO2 + NADPH + H+
-
-
-
r
1.1.1.286
isocitrate + NADP+
-
738195
Sulfolobus acidocaldarius
2-oxoglutarate + CO2 + NADPH
-
-
-
r
1.1.1.286
isocitrate + NADP+
-
738195
Sulfolobus acidocaldarius MW001
2-oxoglutarate + CO2 + NADPH
-
-
-
r
1.1.1.286
additional information
Saci_2375 exhibits distinct and similar activities for isocitrate and homoisocitrate, but no detectable activity for 3-isopropylmalate. Saci_2375 is a dual function enzyme serving as isocitrate-homoisocitrate dehydrogenase
738195
Sulfolobus acidocaldarius
?
-
-
-
-
1.1.1.286
additional information
Saci_2375 exhibits distinct and similar activities for isocitrate and homoisocitrate, but no detectable activity for 3-isopropylmalate. Saci_2375 is a dual function enzyme serving as isocitrate-homoisocitrate dehydrogenase
738195
Sulfolobus acidocaldarius MW001
?
-
-
-
-
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.1.1.85
homotetramer
4 * 36500, SDS-PAGE
Sulfolobus acidocaldarius
1.1.1.286
tetramer
dimer of dimers
Sulfolobus acidocaldarius
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.1.1.286
60
-
assay at
Sulfolobus acidocaldarius
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.1.1.85
0.26
-
homoisocitrate
at pH 8.0 and 60°C
Sulfolobus acidocaldarius
1.1.1.85
8.5
-
(2R,3S)-3-isopropylmalate
at pH 8.0 and 60°C
Sulfolobus acidocaldarius
1.1.1.286
37
-
homoisocitrate
pH 8.0, 60°C, recombinant enzyme
Sulfolobus acidocaldarius
1.1.1.286
61
-
isocitrate
pH 8.0, 60°C, recombinant enzyme
Sulfolobus acidocaldarius
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.286
8
-
assay at
Sulfolobus acidocaldarius
General Information
EC Number
General Information
Commentary
Organism
1.1.1.286
physiological function
the failure to isolate a knockout mutant of saci_2375 suggests the indispensable role of Saci_2375 in cell viability
Sulfolobus acidocaldarius
General Information (protein specific)
EC Number
General Information
Commentary
Organism
1.1.1.286
physiological function
the failure to isolate a knockout mutant of saci_2375 suggests the indispensable role of Saci_2375 in cell viability
Sulfolobus acidocaldarius
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.1.1.85
440
-
homoisocitrate
at pH 8.0 and 60°C
Sulfolobus acidocaldarius
1.1.1.85
1700
-
(2R,3S)-3-isopropylmalate
at pH 8.0 and 60°C
Sulfolobus acidocaldarius
1.1.1.286
2259
-
isocitrate
pH 8.0, 60°C, recombinant enzyme
Sulfolobus acidocaldarius
1.1.1.286
3364
-
homoisocitrate
pH 8.0, 60°C, recombinant enzyme
Sulfolobus acidocaldarius
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.1.1.85
440
-
homoisocitrate
at pH 8.0 and 60°C
Sulfolobus acidocaldarius
1.1.1.85
1700
-
(2R,3S)-3-isopropylmalate
at pH 8.0 and 60°C
Sulfolobus acidocaldarius
1.1.1.286
2259
-
isocitrate
pH 8.0, 60°C, recombinant enzyme
Sulfolobus acidocaldarius
1.1.1.286
3364
-
homoisocitrate
pH 8.0, 60°C, recombinant enzyme
Sulfolobus acidocaldarius