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Literature summary for 1.1.1.100 extracted from

  • Matsumoto, K.; Tanaka, Y.; Watanabe, T.; Motohashi, R.; Ikeda, K.; Tobitani, K.; Yao, M.; Tanaka, I.; Taguchi, S.
    Directed evolution and structural analysis of NADPH-dependent acetoacetyl coenzyme A (acetoacetyl-CoA) reductase from Ralstonia eutropha reveals two mutations responsible for enhanced kinetics (2013), Appl. Environ. Microbiol., 79, 6134-6139.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells and in Corynebacterium glutamicum Cupriavidus necator

Crystallization (Commentary)

Crystallization (Comment) Organism
vapor diffusion method, using 0.1 M MES (pH 7.1), 1.6 M ammonium sulfate, and 10% (w/v) 1,4-dioxane Cupriavidus necator

Protein Variants

Protein Variants Comment Organism
Q47L the mutant with increased specific activity exhibits a kcat value that is 2.4fold higher than that of the wild type enzyme Cupriavidus necator
T173S the mutant with increased specific activity exhibits a kcat value that is 3.5fold higher than that of the wild type enzyme Cupriavidus necator

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0057
-
acetoacetyl-CoA wild type enzyme, at pH 8.0 and 30°C Cupriavidus necator
0.0136
-
acetoacetyl-CoA mutant enzyme Q47L, at pH 8.0 and 30°C Cupriavidus necator
0.0159
-
acetoacetyl-CoA mutant enzyme Q47L, at pH 8.0 and 30°C Cupriavidus necator
0.149
-
NADPH wild type enzyme, at pH 8.0 and 30°C Cupriavidus necator
0.289
-
NADPH mutant enzyme Q47L, at pH 8.0 and 30°C Cupriavidus necator
0.617
-
NADPH mutant enzyme Q47L, at pH 8.0 and 30°C Cupriavidus necator

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
acetoacetyl-CoA + NADPH + H+ Cupriavidus necator
-
3-hydroxybutyryl-CoA + NADP+
-
?
acetoacetyl-CoA + NADPH + H+ Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1
-
3-hydroxybutyryl-CoA + NADP+
-
?

Organism

Organism UniProt Comment Textmining
Cupriavidus necator P14697
-
-
Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 P14697
-
-

Purification (Commentary)

Purification (Comment) Organism
His-Bind resin column chromatography and Superdex 200 pg gel filtration Cupriavidus necator

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
acetoacetyl-CoA + NADPH + H+
-
Cupriavidus necator 3-hydroxybutyryl-CoA + NADP+
-
?
acetoacetyl-CoA + NADPH + H+
-
Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 3-hydroxybutyryl-CoA + NADP+
-
?

Synonyms

Synonyms Comment Organism
NADPH-dependent acetoacetyl coenzyme A reductase
-
Cupriavidus necator
PhaB
-
Cupriavidus necator

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
102
-
NADPH wild type enzyme, at pH 8.0 and 30°C Cupriavidus necator
102
-
acetoacetyl-CoA wild type enzyme, at pH 8.0 and 30°C Cupriavidus necator
249
-
NADPH mutant enzyme Q47L, at pH 8.0 and 30°C Cupriavidus necator
249
-
acetoacetyl-CoA mutant enzyme Q47L, at pH 8.0 and 30°C Cupriavidus necator
361
-
NADPH mutant enzyme Q47L, at pH 8.0 and 30°C Cupriavidus necator
361
-
acetoacetyl-CoA mutant enzyme Q47L, at pH 8.0 and 30°C Cupriavidus necator

Cofactor

Cofactor Comment Organism Structure
NADP+
-
Cupriavidus necator

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
585
-
NADPH mutant enzyme Q47L, at pH 8.0 and 30°C Cupriavidus necator
685
-
NADPH wild type enzyme, at pH 8.0 and 30°C Cupriavidus necator
862
-
NADPH mutant enzyme Q47L, at pH 8.0 and 30°C Cupriavidus necator
15200
-
acetoacetyl-CoA mutant enzyme Q47L, at pH 8.0 and 30°C Cupriavidus necator
18000
-
acetoacetyl-CoA wild type enzyme, at pH 8.0 and 30°C Cupriavidus necator
26500
-
acetoacetyl-CoA mutant enzyme Q47L, at pH 8.0 and 30°C Cupriavidus necator