BRENDA - Enzyme Database

Purification and properties of D-beta-hydroxybutyrate dehydrogenase from Zoogloea ramiger I-16-M

Nakada T.; Fukui, T.; Saito, T.; Miki, K.; Oji, C.; Matsuda, S.; Ushijima, A.; Tomita, K.; J. Biochem. 89, 625-635 (1981)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
1.1.1.30
2-mercaptoethanol
10 mM, 41% increase
Zoogloea ramigera I-16-M
1.1.1.30
dithiothreitol
10 mM, 31% increase
Zoogloea ramigera I-16-M
1.1.1.30
EDTA
0.5 mM, 21% increase
Zoogloea ramigera I-16-M
1.1.1.30
KCN
0.1 mM, 37% increase
Zoogloea ramigera I-16-M
1.1.1.30
phosphatidylcholine
-
Zoogloea ramigera I-16-M
1.1.1.30
reduced glutathione
10 mM, 38% increase
Zoogloea ramigera I-16-M
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
1.1.1.30
2-Hydroxybutyrate
10 mM, 57% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
2-oxobutyrate
10 mM, 29% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
5,5'-dithiobis(2-nitrobenzoic acid)
0.15 mM, 63% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
acetoacetate
inhibition above 1 mM, 2 mM, 89% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
acetoacetyl-CoA
1 mM, 29% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
acetyl-CoA
1 mM, 82% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
Butyrate
10 mM, 20% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
crotonate
10 mM, 20% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
CuSO4
1 mM, 81% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
HgCl2
0.1 mM, 100% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
L-3-hydroxybutyrate
5 mM, 73% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
malonate
10 mM, 66% inhibition, oxidation
Zoogloea ramigera I-16-M
1.1.1.30
MnCl2
1 mM, 50% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
NADH
0.15 mM, 48% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
p-chloromercuribenzoate
0.1 mM, 100% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
pyruvate
4 mM, 5% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
succinate
10 mM, 12% inhibition
Zoogloea ramigera I-16-M
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.30
0.015
-
NADH
-
Zoogloea ramigera I-16-M
1.1.1.30
0.057
-
NAD+
-
Zoogloea ramigera I-16-M
1.1.1.30
0.15
-
acetoacetate
-
Zoogloea ramigera I-16-M
1.1.1.30
0.32
-
beta-hydroxybutyrate
-
Zoogloea ramigera I-16-M
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
1.1.1.30
Ca2+
10 mM 20% increase in activity
Zoogloea ramigera I-16-M
1.1.1.30
Mg2+
10 mM 41% increase in activity
Zoogloea ramigera I-16-M
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.1.30
28000
-
alpha4, 4 * 28000, SDS-PAGE
Zoogloea ramigera I-16-M
1.1.1.30
112000
-
gel filtration
Zoogloea ramigera I-16-M
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.1.1.30
(R)-3-hydroxybutanoate + NAD+
Zoogloea ramigera I-16-M
-
acetoacetate + NADH
-
Zoogloea ramigera I-16-M
r
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.1.1.30
Zoogloea ramigera I-16-M
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.1.1.30
ammonium sulfate, DEAE-Sepharose, 60░C, 10 min, hydroxylapatite, octyl-Sepharose, Sephadex G-200, DEAE-Sepharose
Zoogloea ramigera I-16-M
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [Ámol/min/mg]
Specific Activity Maximum [Ámol/min/mg]
Commentary
Organism
1.1.1.30
123
-
-
Zoogloea ramigera I-16-M
Storage Stability
EC Number
Storage Stability
Organism
1.1.1.30
-20░C, 1 mg enzyme/ml, 0.2 mM potassium phosphate pH 8.0, 10% glycerol, 10 mM 2-mercaptoethanol, 6 months, 0% inactivation, 0░C, 0.1 mg enzyme/ml, 1 day, 50% inactivation
Zoogloea ramigera I-16-M
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.30
(R)-3-hydroxybutanoate + NAD+
-
286534
Zoogloea ramigera I-16-M
acetoacetate + NADH
-
286534
Zoogloea ramigera I-16-M
r
Subunits
EC Number
Subunits
Commentary
Organism
1.1.1.30
tetramer
alpha4, 4 * 28000, SDS-PAGE
Zoogloea ramigera I-16-M
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.30
8
-
oxidation of beta-hydroxybutyrate
Zoogloea ramigera I-16-M
pH Range
EC Number
pH Minimum
pH Maximum
Commentary
Organism
1.1.1.30
5.5
7.5
acetoacetate reduction
Zoogloea ramigera I-16-M
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.30
NAD+
-
Zoogloea ramigera I-16-M
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
1.1.1.30
2-mercaptoethanol
10 mM, 41% increase
Zoogloea ramigera I-16-M
1.1.1.30
dithiothreitol
10 mM, 31% increase
Zoogloea ramigera I-16-M
1.1.1.30
EDTA
0.5 mM, 21% increase
Zoogloea ramigera I-16-M
1.1.1.30
KCN
0.1 mM, 37% increase
Zoogloea ramigera I-16-M
1.1.1.30
phosphatidylcholine
-
Zoogloea ramigera I-16-M
1.1.1.30
reduced glutathione
10 mM, 38% increase
Zoogloea ramigera I-16-M
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.30
NAD+
-
Zoogloea ramigera I-16-M
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.1.1.30
2-Hydroxybutyrate
10 mM, 57% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
2-oxobutyrate
10 mM, 29% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
5,5'-dithiobis(2-nitrobenzoic acid)
0.15 mM, 63% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
acetoacetate
inhibition above 1 mM, 2 mM, 89% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
acetoacetyl-CoA
1 mM, 29% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
acetyl-CoA
1 mM, 82% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
Butyrate
10 mM, 20% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
crotonate
10 mM, 20% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
CuSO4
1 mM, 81% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
HgCl2
0.1 mM, 100% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
L-3-hydroxybutyrate
5 mM, 73% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
malonate
10 mM, 66% inhibition, oxidation
Zoogloea ramigera I-16-M
1.1.1.30
MnCl2
1 mM, 50% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
NADH
0.15 mM, 48% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
p-chloromercuribenzoate
0.1 mM, 100% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
pyruvate
4 mM, 5% inhibition
Zoogloea ramigera I-16-M
1.1.1.30
succinate
10 mM, 12% inhibition
Zoogloea ramigera I-16-M
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.30
0.015
-
NADH
-
Zoogloea ramigera I-16-M
1.1.1.30
0.057
-
NAD+
-
Zoogloea ramigera I-16-M
1.1.1.30
0.15
-
acetoacetate
-
Zoogloea ramigera I-16-M
1.1.1.30
0.32
-
beta-hydroxybutyrate
-
Zoogloea ramigera I-16-M
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
1.1.1.30
Ca2+
10 mM 20% increase in activity
Zoogloea ramigera I-16-M
1.1.1.30
Mg2+
10 mM 41% increase in activity
Zoogloea ramigera I-16-M
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.1.30
28000
-
alpha4, 4 * 28000, SDS-PAGE
Zoogloea ramigera I-16-M
1.1.1.30
112000
-
gel filtration
Zoogloea ramigera I-16-M
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.1.1.30
(R)-3-hydroxybutanoate + NAD+
Zoogloea ramigera I-16-M
-
acetoacetate + NADH
-
Zoogloea ramigera I-16-M
r
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.30
ammonium sulfate, DEAE-Sepharose, 60░C, 10 min, hydroxylapatite, octyl-Sepharose, Sephadex G-200, DEAE-Sepharose
Zoogloea ramigera I-16-M
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [Ámol/min/mg]
Specific Activity Maximum [Ámol/min/mg]
Commentary
Organism
1.1.1.30
123
-
-
Zoogloea ramigera I-16-M
Storage Stability (protein specific)
EC Number
Storage Stability
Organism
1.1.1.30
-20░C, 1 mg enzyme/ml, 0.2 mM potassium phosphate pH 8.0, 10% glycerol, 10 mM 2-mercaptoethanol, 6 months, 0% inactivation, 0░C, 0.1 mg enzyme/ml, 1 day, 50% inactivation
Zoogloea ramigera I-16-M
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.30
(R)-3-hydroxybutanoate + NAD+
-
286534
Zoogloea ramigera I-16-M
acetoacetate + NADH
-
286534
Zoogloea ramigera I-16-M
r
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.1.1.30
tetramer
alpha4, 4 * 28000, SDS-PAGE
Zoogloea ramigera I-16-M
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.30
8
-
oxidation of beta-hydroxybutyrate
Zoogloea ramigera I-16-M
pH Range (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
Organism
1.1.1.30
5.5
7.5
acetoacetate reduction
Zoogloea ramigera I-16-M