BRENDA - Enzyme Database

Purification and characterization of NADPH-dependent acetoacetyl-CoA reductase from Methylobacterium extorquens

Belova, L.L.; Sokolov, A.P.; Sidorov, I.A.; Trotsenko, Y.A.; FEMS Microbiol. Lett. 156, 275-279 (1997)
No PubMed abstract available

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
1.1.1.36
alpha-ketoglutarate
20% activation
Methylorubrum extorquens
1.1.1.36
ATP
32% activation
Methylorubrum extorquens
1.1.1.36
citrate
30% activation
Methylorubrum extorquens
1.1.1.36
isocitrate
25% activation, stimulates NADP+ regeneration
Methylorubrum extorquens
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
1.1.1.36
acetoacetyl-CoA
-
Azotobacter vinelandii
1.1.1.36
acetoacetyl-CoA
-
Cupriavidus necator
1.1.1.36
acetoacetyl-CoA
concentration above 40 mM
Methylorubrum extorquens
1.1.1.36
acetoacetyl-CoA
-
Methylorubrum rhodesianum
1.1.1.36
acetoacetyl-CoA
-
Zoogloea ramigera
1.1.1.36
NAD+
17% inhibition
Methylorubrum extorquens
1.1.1.36
NAD+
-
Methylorubrum rhodesianum
1.1.1.36
NADP+
40% inhibition
Methylorubrum extorquens
1.1.1.36
NADP+
-
Methylorubrum rhodesianum
1.1.1.36
NADPH
-
Azotobacter vinelandii
1.1.1.36
NADPH
concentrationgreater 100 mM
Methylorubrum extorquens
1.1.1.36
NADPH
-
Methylorubrum rhodesianum
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.36
0.005
-
acetoacetyl-CoA
-
Cupriavidus necator
1.1.1.36
0.011
-
acetoacetyl-CoA
100 mM NADPH
Azotobacter vinelandii
1.1.1.36
0.0116
-
acetoacetyl-CoA
-
Methylorubrum extorquens
1.1.1.36
0.015
-
acetoacetyl-CoA
-
Methylorubrum rhodesianum
1.1.1.36
0.018
-
NADPH
-
Methylorubrum rhodesianum
1.1.1.36
0.019
-
NADPH
-
Cupriavidus necator
1.1.1.36
0.02
-
NADPH
32 mM acetoacetyl-CoA
Azotobacter vinelandii
1.1.1.36
0.03
-
D-beta-hydroxybutyryl-CoA
-
Methylorubrum rhodesianum
1.1.1.36
0.031
-
NADP+
-
Cupriavidus necator
1.1.1.36
0.033
-
D-hydroxybutyryl-CoA
-
Cupriavidus necator
1.1.1.36
0.041
-
NADPH
-
Methylorubrum extorquens
1.1.1.36
0.06
-
NADP+
-
Methylorubrum rhodesianum
1.1.1.36
0.25
-
NAD+
-
Cupriavidus necator
1.1.1.36
0.4
-
NADH
-
Cupriavidus necator
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
1.1.1.36
soluble
-
Methylorubrum extorquens
-
-
1.1.1.36
soluble
-
Zoogloea ramigera
-
-
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.1.36
31000
-
4 * 31000, 12% PAGE
Methylorubrum extorquens
1.1.1.36
77000
-
-
Azotobacter vinelandii
1.1.1.36
84000
-
-
Cupriavidus necator
1.1.1.36
92000
-
-
Zoogloea ramigera
1.1.1.36
140000
-
gel-filtration
Methylorubrum extorquens
1.1.1.36
141000
-
gradient PAGE
Methylorubrum extorquens
1.1.1.36
250000
-
-
Methylorubrum rhodesianum
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.1.1.36
acetoacetyl-CoA + NADH
Zoogloea ramigera
-
L-3-hydroxybutyryl-CoA + NAD+
-
Zoogloea ramigera
?
1.1.1.36
acetoacetyl-CoA + NADH
Rhodospirillum rubrum
-
L-3-hydroxybutyryl-CoA + NAD+
-
Rhodospirillum rubrum
?
1.1.1.36
acetoacetyl-CoA + NADPH
Zoogloea ramigera
PHB synthesis
D-3-hydroxybutyryl-CoA + NADP+
-
Zoogloea ramigera
r
1.1.1.36
acetoacetyl-CoA + NADPH
Cupriavidus necator
PHB synthesis
D-3-hydroxybutyryl-CoA + NADP+
-
Cupriavidus necator
-
1.1.1.36
acetoacetyl-CoA + NADPH
Methylorubrum extorquens
tricarboxylic acid cycle, poly-3-hydroxybutyrate synthesis, aerobic serine pathway
D-3-hydroxybutyryl-CoA + NADP+
-
Methylorubrum extorquens
r
1.1.1.36
acetoacetyl-CoA + NADPH
Methylorubrum extorquens 15
tricarboxylic acid cycle, poly-3-hydroxybutyrate synthesis, aerobic serine pathway
D-3-hydroxybutyryl-CoA + NADP+
-
Methylorubrum extorquens 15
r
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.1.1.36
Azotobacter vinelandii
-
-
-
1.1.1.36
Cupriavidus necator
-
-
-
1.1.1.36
Methylorubrum extorquens
-
15
-
1.1.1.36
Methylorubrum extorquens 15
-
15
-
1.1.1.36
Methylorubrum rhodesianum
-
MB 126
-
1.1.1.36
Methylorubrum rhodesianum MB 126
-
MB 126
-
1.1.1.36
Rhodospirillum rubrum
-
-
-
1.1.1.36
Zoogloea ramigera
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.1.1.36
PEG, DEAE-Toyopearl column
Methylorubrum extorquens
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [Ámol/min/mg]
Specific Activity Maximum [Ámol/min/mg]
Commentary
Organism
1.1.1.36
184
-
-
Methylorubrum extorquens
Storage Stability
EC Number
Storage Stability
Organism
1.1.1.36
0░C, 25 mM Tris-HCl, pH 8.0
Methylorubrum extorquens
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.36
acetoacetyl-CoA + NADH
-
286622
Zoogloea ramigera
L-3-hydroxybutyryl-CoA + NAD+
-
286622
Zoogloea ramigera
?
1.1.1.36
acetoacetyl-CoA + NADH
-
286622
Cupriavidus necator
L-3-hydroxybutyryl-CoA + NAD+
-
-
-
-
1.1.1.36
acetoacetyl-CoA + NADH
-
286622
Rhodospirillum rubrum
L-3-hydroxybutyryl-CoA + NAD+
-
286622
Rhodospirillum rubrum
?
1.1.1.36
acetoacetyl-CoA + NADPH
-
286622
Methylorubrum rhodesianum
D-beta-hydroxybutyryl-CoA + NADP+
-
-
-
-
1.1.1.36
acetoacetyl-CoA + NADPH
-
286622
Methylorubrum rhodesianum MB 126
D-beta-hydroxybutyryl-CoA + NADP+
-
-
-
-
1.1.1.36
acetoacetyl-CoA + NADPH
PHB synthesis
286622
Zoogloea ramigera
D-3-hydroxybutyryl-CoA + NADP+
-
286622
Zoogloea ramigera
r
1.1.1.36
acetoacetyl-CoA + NADPH
PHB synthesis
286622
Cupriavidus necator
D-3-hydroxybutyryl-CoA + NADP+
-
286622
Cupriavidus necator
-
1.1.1.36
acetoacetyl-CoA + NADPH
tricarboxylic acid cycle, poly-3-hydroxybutyrate synthesis, aerobic serine pathway
286622
Methylorubrum extorquens
D-3-hydroxybutyryl-CoA + NADP+
-
286622
Methylorubrum extorquens
r
1.1.1.36
acetoacetyl-CoA + NADPH
tricarboxylic acid cycle, poly-3-hydroxybutyrate synthesis, aerobic serine pathway
286622
Methylorubrum extorquens 15
D-3-hydroxybutyryl-CoA + NADP+
-
286622
Methylorubrum extorquens 15
r
Subunits
EC Number
Subunits
Commentary
Organism
1.1.1.36
tetramer
4 * 31000, 12% PAGE
Methylorubrum extorquens
1.1.1.36
tetramer
-
Zoogloea ramigera
Temperature Optimum [░C]
EC Number
Temperature Optimum [░C]
Temperature Optimum Maximum [░C]
Commentary
Organism
1.1.1.36
30
-
-
Methylorubrum extorquens
1.1.1.36
30
-
-
Zoogloea ramigera
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.36
7.5
-
-
Methylorubrum extorquens
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.36
NADPH
-
Azotobacter vinelandii
1.1.1.36
NADPH
absolute specificity
Methylorubrum extorquens
1.1.1.36
NADPH
absolute specificity
Zoogloea ramigera
1.1.1.36
NADPH
-
Methylorubrum rhodesianum
1.1.1.36
NADPH
-
Cupriavidus necator
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
1.1.1.36
alpha-ketoglutarate
20% activation
Methylorubrum extorquens
1.1.1.36
ATP
32% activation
Methylorubrum extorquens
1.1.1.36
citrate
30% activation
Methylorubrum extorquens
1.1.1.36
isocitrate
25% activation, stimulates NADP+ regeneration
Methylorubrum extorquens
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.36
NADPH
-
Azotobacter vinelandii
1.1.1.36
NADPH
absolute specificity
Methylorubrum extorquens
1.1.1.36
NADPH
absolute specificity
Zoogloea ramigera
1.1.1.36
NADPH
-
Methylorubrum rhodesianum
1.1.1.36
NADPH
-
Cupriavidus necator
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.1.1.36
acetoacetyl-CoA
-
Azotobacter vinelandii
1.1.1.36
acetoacetyl-CoA
-
Cupriavidus necator
1.1.1.36
acetoacetyl-CoA
concentration above 40 mM
Methylorubrum extorquens
1.1.1.36
acetoacetyl-CoA
-
Methylorubrum rhodesianum
1.1.1.36
acetoacetyl-CoA
-
Zoogloea ramigera
1.1.1.36
NAD+
17% inhibition
Methylorubrum extorquens
1.1.1.36
NAD+
-
Methylorubrum rhodesianum
1.1.1.36
NADP+
40% inhibition
Methylorubrum extorquens
1.1.1.36
NADP+
-
Methylorubrum rhodesianum
1.1.1.36
NADPH
-
Azotobacter vinelandii
1.1.1.36
NADPH
concentrationgreater 100 mM
Methylorubrum extorquens
1.1.1.36
NADPH
-
Methylorubrum rhodesianum
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.36
0.005
-
acetoacetyl-CoA
-
Cupriavidus necator
1.1.1.36
0.011
-
acetoacetyl-CoA
100 mM NADPH
Azotobacter vinelandii
1.1.1.36
0.0116
-
acetoacetyl-CoA
-
Methylorubrum extorquens
1.1.1.36
0.015
-
acetoacetyl-CoA
-
Methylorubrum rhodesianum
1.1.1.36
0.018
-
NADPH
-
Methylorubrum rhodesianum
1.1.1.36
0.019
-
NADPH
-
Cupriavidus necator
1.1.1.36
0.02
-
NADPH
32 mM acetoacetyl-CoA
Azotobacter vinelandii
1.1.1.36
0.03
-
D-beta-hydroxybutyryl-CoA
-
Methylorubrum rhodesianum
1.1.1.36
0.031
-
NADP+
-
Cupriavidus necator
1.1.1.36
0.033
-
D-hydroxybutyryl-CoA
-
Cupriavidus necator
1.1.1.36
0.041
-
NADPH
-
Methylorubrum extorquens
1.1.1.36
0.06
-
NADP+
-
Methylorubrum rhodesianum
1.1.1.36
0.25
-
NAD+
-
Cupriavidus necator
1.1.1.36
0.4
-
NADH
-
Cupriavidus necator
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
1.1.1.36
soluble
-
Methylorubrum extorquens
-
-
1.1.1.36
soluble
-
Zoogloea ramigera
-
-
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.1.36
31000
-
4 * 31000, 12% PAGE
Methylorubrum extorquens
1.1.1.36
77000
-
-
Azotobacter vinelandii
1.1.1.36
84000
-
-
Cupriavidus necator
1.1.1.36
92000
-
-
Zoogloea ramigera
1.1.1.36
140000
-
gel-filtration
Methylorubrum extorquens
1.1.1.36
141000
-
gradient PAGE
Methylorubrum extorquens
1.1.1.36
250000
-
-
Methylorubrum rhodesianum
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.1.1.36
acetoacetyl-CoA + NADH
Zoogloea ramigera
-
L-3-hydroxybutyryl-CoA + NAD+
-
Zoogloea ramigera
?
1.1.1.36
acetoacetyl-CoA + NADH
Rhodospirillum rubrum
-
L-3-hydroxybutyryl-CoA + NAD+
-
Rhodospirillum rubrum
?
1.1.1.36
acetoacetyl-CoA + NADPH
Zoogloea ramigera
PHB synthesis
D-3-hydroxybutyryl-CoA + NADP+
-
Zoogloea ramigera
r
1.1.1.36
acetoacetyl-CoA + NADPH
Cupriavidus necator
PHB synthesis
D-3-hydroxybutyryl-CoA + NADP+
-
Cupriavidus necator
-
1.1.1.36
acetoacetyl-CoA + NADPH
Methylorubrum extorquens
tricarboxylic acid cycle, poly-3-hydroxybutyrate synthesis, aerobic serine pathway
D-3-hydroxybutyryl-CoA + NADP+
-
Methylorubrum extorquens
r
1.1.1.36
acetoacetyl-CoA + NADPH
Methylorubrum extorquens 15
tricarboxylic acid cycle, poly-3-hydroxybutyrate synthesis, aerobic serine pathway
D-3-hydroxybutyryl-CoA + NADP+
-
Methylorubrum extorquens 15
r
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.36
PEG, DEAE-Toyopearl column
Methylorubrum extorquens
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [Ámol/min/mg]
Specific Activity Maximum [Ámol/min/mg]
Commentary
Organism
1.1.1.36
184
-
-
Methylorubrum extorquens
Storage Stability (protein specific)
EC Number
Storage Stability
Organism
1.1.1.36
0░C, 25 mM Tris-HCl, pH 8.0
Methylorubrum extorquens
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.36
acetoacetyl-CoA + NADH
-
286622
Zoogloea ramigera
L-3-hydroxybutyryl-CoA + NAD+
-
286622
Zoogloea ramigera
?
1.1.1.36
acetoacetyl-CoA + NADH
-
286622
Cupriavidus necator
L-3-hydroxybutyryl-CoA + NAD+
-
-
-
-
1.1.1.36
acetoacetyl-CoA + NADH
-
286622
Rhodospirillum rubrum
L-3-hydroxybutyryl-CoA + NAD+
-
286622
Rhodospirillum rubrum
?
1.1.1.36
acetoacetyl-CoA + NADPH
-
286622
Methylorubrum rhodesianum
D-beta-hydroxybutyryl-CoA + NADP+
-
-
-
-
1.1.1.36
acetoacetyl-CoA + NADPH
-
286622
Methylorubrum rhodesianum MB 126
D-beta-hydroxybutyryl-CoA + NADP+
-
-
-
-
1.1.1.36
acetoacetyl-CoA + NADPH
PHB synthesis
286622
Zoogloea ramigera
D-3-hydroxybutyryl-CoA + NADP+
-
286622
Zoogloea ramigera
r
1.1.1.36
acetoacetyl-CoA + NADPH
PHB synthesis
286622
Cupriavidus necator
D-3-hydroxybutyryl-CoA + NADP+
-
286622
Cupriavidus necator
-
1.1.1.36
acetoacetyl-CoA + NADPH
tricarboxylic acid cycle, poly-3-hydroxybutyrate synthesis, aerobic serine pathway
286622
Methylorubrum extorquens
D-3-hydroxybutyryl-CoA + NADP+
-
286622
Methylorubrum extorquens
r
1.1.1.36
acetoacetyl-CoA + NADPH
tricarboxylic acid cycle, poly-3-hydroxybutyrate synthesis, aerobic serine pathway
286622
Methylorubrum extorquens 15
D-3-hydroxybutyryl-CoA + NADP+
-
286622
Methylorubrum extorquens 15
r
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.1.1.36
tetramer
4 * 31000, 12% PAGE
Methylorubrum extorquens
1.1.1.36
tetramer
-
Zoogloea ramigera
Temperature Optimum [░C] (protein specific)
EC Number
Temperature Optimum [░C]
Temperature Optimum Maximum [░C]
Commentary
Organism
1.1.1.36
30
-
-
Methylorubrum extorquens
1.1.1.36
30
-
-
Zoogloea ramigera
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.36
7.5
-
-
Methylorubrum extorquens