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Literature summary extracted from

  • Takahashi, K.; Nakanishi, F.; Tomita, T.; Akiyama, N.; Lassak, K.; Albers, S.V.; Kuzuyama, T.; Nishiyama, M.
    Characterization of two ?-decarboxylating dehydrogenases from Sulfolobus acidocaldarius (2016), Extremophiles, 20, 843-853.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) 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 (Comment) 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 Comment 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 Comment Organism Structure
1.1.1.85 Mg2+ required Sulfolobus acidocaldarius

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment 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 Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
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 + H+
-
 r
1.1.1.286 isocitrate + NADP+ Sulfolobus acidocaldarius MW001
-
 2-oxoglutarate + CO2 + NADPH + H+
-
 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 UniProt Comment 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 Purification (Comment) 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 Comment Substrates Organism Products Comment (Products) Rev. Reac.
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+
-
 ?
1.1.1.85 homoisocitrate + NAD+
-
 Sulfolobus acidocaldarius ? + NADH + H+
-
 ?
1.1.1.85 homoisocitrate + NAD+
-
 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+ 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+ Sulfolobus acidocaldarius MW001 ?
-
 ?
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 + H+
-
 r
1.1.1.286 isocitrate + NADP+
-
 Sulfolobus acidocaldarius MW001 2-oxoglutarate + CO2 + NADPH + H+
-
 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 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 Sulfolobus acidocaldarius MW001 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.85 homotetramer 4 * 36500, SDS-PAGE Sulfolobus acidocaldarius
1.1.1.286 tetramer dimer of dimers Sulfolobus acidocaldarius

Synonyms

EC Number Synonyms Comment Organism
1.1.1.85 IPMDH
-
 Sulfolobus acidocaldarius
1.1.1.85 Saci_0600
-
 Sulfolobus acidocaldarius
1.1.1.286 isocitrate-homoisocitrate dehydrogenase
-
 Sulfolobus acidocaldarius
1.1.1.286 Saci_2375
-
 Sulfolobus acidocaldarius
1.1.1.286 Saci_2375 locus name Sulfolobus acidocaldarius

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment 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 Comment 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 Comment Organism
1.1.1.286 8
-
 assay at Sulfolobus acidocaldarius

Cofactor

EC Number Cofactor Comment 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

General Information

EC Number General Information Comment 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 Comment 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