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Literature summary extracted from

  • Suzuki, Y.; Asada, K.; Miyazaki, J.; Tomita, T.; Kuzuyama, T.; Nishiyama, M.
    Enhancement of latent 3-isopropylmalate dehydrogenase activity of promiscuous homoisocitrate dehydrogenase by directed evolution (2010), Biochem. J., 431, 401-410.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.85 expressed in Escherichia coli Thermus thermophilus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.1.1.85 crystal structure of mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M is determined at 2.4 A Thermus thermophilus

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.85 E57V/R85M/Y86A/M208T/F217Y/V238M/R310M no (2R,3S)-3-isopropylmalate dehydrogenase activity detected Thermus thermophilus
1.1.1.85 E57V/S72I/R85M/Y86A/F217Y/V238M/R310M no (2R,3S)-3-isopropylmalate dehydrogenase activity detected Thermus thermophilus
1.1.1.85 E57V/S72I/R85M/Y86A/M208T/F217Y/R310M in comparison to mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M this mutant shows a significant decrease in kcat ((2R,3S)-3-isopropylmalate), alteration of Km value only moderate Thermus thermophilus
1.1.1.85 E57V/S72I/R85M/Y86A/M208T/F217Y/V238M in comparison to mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M this mutant shows a significant decrease in kcat ((2R,3S)-3-isopropylmalate), alteration of Km value only moderate Thermus thermophilus
1.1.1.85 E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M in comparison to mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M this mutant shows a larger kcat ((2R,3S)-3-isopropylmalate) and a larger Km((2R,3S)-3-isopropylmalate) or (NAD+) temperatures giving half of the initial activity is 87.8 °C Thermus thermophilus
1.1.1.85 E57V/S72I/R85M/Y86A/M208T/V238M/R310M in comparison to mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M this mutant shows a significant decrease in kcat ((2R,3S)-3-isopropylmalate), alteration of Km value only moderate Thermus thermophilus
1.1.1.85 E57V/S72I/R85V/Y86A/M208T/F217Y/V238M/R310M no (2R,3S)-3-isopropylmalate dehydrogenase activity detected Thermus thermophilus
1.1.1.85 H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M TtHICDH variant with considerably higher IPMDH activity than double mutant R85V/Y86A, Km ((2R,3S)-3-isopropylmalate) or (NAD+) lower than wild-type HICDH but still considerably higher than wild-type (2R,3S)-3-isopropylmalate dehydrogenase, kcat ((2R,3S)-3-isopropylmalate) much higher than wild-type HICDH but still considerably lower than wild-type (2R,3S)-3-isopropylmalate dehydrogenase, temperatures giving half of the initial activity is 79.9 °C, crystal structure determined at 2.4 A, mutant is a homotetramer Thermus thermophilus
1.1.1.85 R85V/Y86A TtHICDH variant with high IPMDH activity, Km ((2R,3S)-3-isopropylmalate) or (NAD+) lower than wild-type HICDH but still considerably higher than wild-type (2R,3S)-3-isopropylmalate dehydrogenase, kcat ((2R,3S)-3-isopropylmalate) higher than wild-type HICDH but still considerably lower than wild-type (2R,3S)-3-isopropylmalate dehydrogenase, temperatures giving half of the initial activity is 92.5 °C Thermus thermophilus
1.1.1.85 S72I/R85M/Y86A/M208T/F217Y/V238M/R310M in comparison to mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M this mutant shows a significant decrease in kcat ((2R,3S)-3-isopropylmalate), alteration of Km value only moderate Thermus thermophilus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.85 0.59
-
(2R,3S)-3-isopropylmalate mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 0.9
-
(2R,3S)-3-isopropylmalate mutant E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 0.9
-
NAD+ mutant E57V/S72I/R85M/Y86A/M208T/V238M/R310M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 0.9
-
(2R,3S)-3-isopropylmalate mutant E57V/S72I/R85M/Y86A/M208T/V238M/R310M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 1
-
(2R,3S)-3-isopropylmalate mutant S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 1
-
(2R,3S)-3-isopropylmalate wild-type (2R,3S)-3-isopropylmalate dehydrogenase, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 1
-
(2R,3S)-3-isopropylmalate wild-type HICDH, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 1.2
-
NAD+ mutant E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 1.2
-
NAD+ mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 1.5
-
(2R,3S)-3-isopropylmalate mutant E57V/S72I/R85M/Y86A/M208T/F217Y/V238M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 1.6
-
NAD+ mutant E57V/S72I/R85M/Y86A/M208T/F217Y/V238M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 1.6
-
NAD+ mutant S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 1.7
-
NAD+ mutant E57V/S72I/R85M/Y86A/M208T/F217Y/R310M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 1.7
-
NAD+ mutant R85V/Y86A, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 1.9
-
NAD+ wild-type (2R,3S)-3-isopropylmalate dehydrogenase, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 1.9
-
NAD+ wild-type HICDH, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 1.9
-
(2R,3S)-3-isopropylmalate mutant E57V/S72I/R85M/Y86A/M208T/F217Y/R310M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 2.6
-
(2R,3S)-3-isopropylmalate mutant R85V/Y86A, pH 8.0, 37°C Thermus thermophilus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.85 Mg2+
-
Thermus thermophilus

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.85 Thermus thermophilus Q72IW9
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.85 (2R,3S)-3-isopropylmalate + NAD+ mutant enzymes with increased activity Thermus thermophilus (2S)-2-isopropyl-3-oxosuccinate + NADH + H+
-
?

Synonyms

EC Number Synonyms Comment Organism
1.1.1.85 IPMDH
-
Thermus thermophilus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.85 37
-
assay at Thermus thermophilus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.85 0.08
-
(2R,3S)-3-isopropylmalate wild-type HICDH, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 0.3
-
(2R,3S)-3-isopropylmalate mutant E57V/S72I/R85M/Y86A/M208T/V238M/R310M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 0.4
-
(2R,3S)-3-isopropylmalate mutant R85V/Y86A, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 0.6
-
(2R,3S)-3-isopropylmalate mutant E57V/S72I/R85M/Y86A/M208T/F217Y/V238M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 1.6
-
(2R,3S)-3-isopropylmalate mutant S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 1.8
-
(2R,3S)-3-isopropylmalate mutant E57V/S72I/R85M/Y86A/M208T/F217Y/R310M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 2.9
-
(2R,3S)-3-isopropylmalate mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 5.1
-
(2R,3S)-3-isopropylmalate mutant E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 5.7
-
(2R,3S)-3-isopropylmalate wild-type (2R,3S)-3-isopropylmalate dehydrogenase, pH 8.0, 37°C Thermus thermophilus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.85 8
-
assay at Thermus thermophilus

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.85 NADH
-
Thermus thermophilus

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 0.12
-
NAD+ wild-type (2R,3S)-3-isopropylmalate dehydrogenase, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 0.16
-
(2R,3S)-3-isopropylmalate mutant R85V/Y86A, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 0.18
-
(2R,3S)-3-isopropylmalate wild-type (2R,3S)-3-isopropylmalate dehydrogenase, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 0.23
-
NAD+ mutant R85V/Y86A, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 0.25
-
NAD+ mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 0.33
-
NAD+ mutant E57V/S72I/R85M/Y86A/M208T/V238M/R310M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 0.33
-
(2R,3S)-3-isopropylmalate mutant E57V/S72I/R85M/Y86A/M208T/V238M/R310M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 0.38
-
NAD+ mutant E57V/S72I/R85M/Y86A/M208T/F217Y/V238M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 0.4
-
(2R,3S)-3-isopropylmalate mutant E57V/S72I/R85M/Y86A/M208T/F217Y/V238M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 0.41
-
NAD+ wild-type HICDH, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 0.49
-
(2R,3S)-3-isopropylmalate mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 0.75
-
(2R,3S)-3-isopropylmalate wild-type HICDH, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 0.94
-
(2R,3S)-3-isopropylmalate mutant E57V/S72I/R85M/Y86A/M208T/F217Y/R310M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 1.1
-
NAD+ mutant E57V/S72I/R85M/Y86A/M208T/F217Y/R310M , pH 8.0, 37°C Thermus thermophilus
1.1.1.85 1.4
-
NAD+ mutant S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 1.6
-
(2R,3S)-3-isopropylmalate mutant S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 4.2
-
NAD+ mutant E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C Thermus thermophilus
1.1.1.85 5.4
-
(2R,3S)-3-isopropylmalate mutant E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C Thermus thermophilus