BRENDA - Enzyme Database show
show all sequences of 1.1.99.6

A novel flavin adenine dinucleotide (FAD) containing D-lactate dehydrogenase from the thermoacidophilic crenarchaeota Sulfolobus tokodaii strain 7: purification, characterization and expression in Escherichia coli

Satomura, T.; Kawakami, R.; Sakuraba, H.; Ohshima, T.; J. Biosci. Bioeng. 106, 16-21 (2008)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expression in Escherichia coli
Sulfurisphaera tokodaii
Inhibitors
Inhibitors
Commentary
Organism
Structure
4-chloromercuribenzoate
1 mM, 62% loss of activity
Sulfurisphaera tokodaii
CoCl2
1 mM, 78% loss of activity
Sulfurisphaera tokodaii
CuCl2
1 mM, complete loss of activity
Sulfurisphaera tokodaii
HgCl2
1 mM, 93% loss of activity
Sulfurisphaera tokodaii
MnCl2
1 mM, complete loss of activity
Sulfurisphaera tokodaii
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.091
-
(R)-lactate
pH 8.5, 60°C, cosubstrate: 2,6-dichlorophenolindophenol
Sulfurisphaera tokodaii
0.156
-
(R)-lactate
pH 8.5, 60°C, cosubstrate: 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide
Sulfurisphaera tokodaii
0.247
-
oxidized 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide
pH 8.5, 60°C, cosubstrate: (R)-lactate
Sulfurisphaera tokodaii
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
48000
-
2 * 48000, SDS-PAGE
Sulfurisphaera tokodaii
49715
-
2 * 49715, calculated from sequence
Sulfurisphaera tokodaii
93000
-
non-denaturing PAGE
Sulfurisphaera tokodaii
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Sulfurisphaera tokodaii
Q974L0
-
-
Sulfurisphaera tokodaii 7
Q974L0
-
-
Purification (Commentary)
Commentary
Organism
-
Sulfurisphaera tokodaii
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(R)-2-hydroxybutyrate + oxidized 2,6-dichlorophenolindophenol
97% of the activity compared to (R)-lactate
693285
Sulfurisphaera tokodaii
2-oxobutyrate + reduced 2,6-dichlorophenolindophenol
-
-
-
?
(R)-2-hydroxybutyrate + oxidized 2,6-dichlorophenolindophenol
97% of the activity compared to (R)-lactate
693285
Sulfurisphaera tokodaii 7
2-oxobutyrate + reduced 2,6-dichlorophenolindophenol
-
-
-
?
(R)-lactate + ferricyanide
ferricyanide is the preferred electron acceptor
693285
Sulfurisphaera tokodaii
pyruvate + ferrocyanide
-
-
-
?
(R)-lactate + ferricyanide
ferricyanide is the preferred electron acceptor
693285
Sulfurisphaera tokodaii 7
pyruvate + ferrocyanide
-
-
-
?
(R)-lactate + oxidized 2,6-dichlorophenolindophenol
activity is 59% compared to the activity with ferricyanide
693285
Sulfurisphaera tokodaii
pyruvate + reduced 2,6-dichlorophenolindophenol
-
-
-
?
(R)-lactate + oxidized 2,6-dichlorophenolindophenol
activity is 59% compared to the activity with ferricyanide
693285
Sulfurisphaera tokodaii 7
pyruvate + reduced 2,6-dichlorophenolindophenol
-
-
-
?
(R)-lactate + oxidized 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide
activity is 29% compared to the activity with ferricyanide
693285
Sulfurisphaera tokodaii
pyruvate + reduced 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide
-
-
-
?
(R)-lactate + oxidized 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide
activity is 29% compared to the activity with ferricyanide
693285
Sulfurisphaera tokodaii 7
pyruvate + reduced 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide
-
-
-
?
additional information
L-lactate, glycolate, L-tartrate, citrate, glyoxylate, 2-hydroxyisobutyrate, D-glycerate and D-2-phosphoglycerate are inert as electron donors. Benzyl viologen and methylene blue are inert as electron acceptors
693285
Sulfurisphaera tokodaii
?
-
-
-
-
additional information
L-lactate, glycolate, L-tartrate, citrate, glyoxylate, 2-hydroxyisobutyrate, D-glycerate and D-2-phosphoglycerate are inert as electron donors. Benzyl viologen and methylene blue are inert as electron acceptors
693285
Sulfurisphaera tokodaii 7
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
homodimer
2 * 48000, SDS-PAGE; 2 * 49715, calculated from sequence
Sulfurisphaera tokodaii
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
60
-
assay at
Sulfurisphaera tokodaii
80
-
-
Sulfurisphaera tokodaii
Temperature Stability [°C]
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
80
-
10 min, enzyme retains full activity
Sulfurisphaera tokodaii
85
-
10 min, more than 60% loss of activity
Sulfurisphaera tokodaii
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8.5
-
60°C
Sulfurisphaera tokodaii
pH Range
pH Minimum
pH Maximum
Commentary
Organism
6.5
9
pH 6.5: about 50% of maximal activity, pH 9.0: about 50% of maximal activity
Sulfurisphaera tokodaii
pH Stability
pH Stability
pH Stability Maximum
Commentary
Organism
6.5
10
50°C, 30 min, enzyme retains full activity
Sulfurisphaera tokodaii
Cofactor
Cofactor
Commentary
Organism
Structure
FAD
prosthetic group
Sulfurisphaera tokodaii
Cloned(Commentary) (protein specific)
Commentary
Organism
expression in Escherichia coli
Sulfurisphaera tokodaii
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
FAD
prosthetic group
Sulfurisphaera tokodaii
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
4-chloromercuribenzoate
1 mM, 62% loss of activity
Sulfurisphaera tokodaii
CoCl2
1 mM, 78% loss of activity
Sulfurisphaera tokodaii
CuCl2
1 mM, complete loss of activity
Sulfurisphaera tokodaii
HgCl2
1 mM, 93% loss of activity
Sulfurisphaera tokodaii
MnCl2
1 mM, complete loss of activity
Sulfurisphaera tokodaii
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.091
-
(R)-lactate
pH 8.5, 60°C, cosubstrate: 2,6-dichlorophenolindophenol
Sulfurisphaera tokodaii
0.156
-
(R)-lactate
pH 8.5, 60°C, cosubstrate: 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide
Sulfurisphaera tokodaii
0.247
-
oxidized 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide
pH 8.5, 60°C, cosubstrate: (R)-lactate
Sulfurisphaera tokodaii
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
48000
-
2 * 48000, SDS-PAGE
Sulfurisphaera tokodaii
49715
-
2 * 49715, calculated from sequence
Sulfurisphaera tokodaii
93000
-
non-denaturing PAGE
Sulfurisphaera tokodaii
Purification (Commentary) (protein specific)
Commentary
Organism
-
Sulfurisphaera tokodaii
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(R)-2-hydroxybutyrate + oxidized 2,6-dichlorophenolindophenol
97% of the activity compared to (R)-lactate
693285
Sulfurisphaera tokodaii
2-oxobutyrate + reduced 2,6-dichlorophenolindophenol
-
-
-
?
(R)-2-hydroxybutyrate + oxidized 2,6-dichlorophenolindophenol
97% of the activity compared to (R)-lactate
693285
Sulfurisphaera tokodaii 7
2-oxobutyrate + reduced 2,6-dichlorophenolindophenol
-
-
-
?
(R)-lactate + ferricyanide
ferricyanide is the preferred electron acceptor
693285
Sulfurisphaera tokodaii
pyruvate + ferrocyanide
-
-
-
?
(R)-lactate + ferricyanide
ferricyanide is the preferred electron acceptor
693285
Sulfurisphaera tokodaii 7
pyruvate + ferrocyanide
-
-
-
?
(R)-lactate + oxidized 2,6-dichlorophenolindophenol
activity is 59% compared to the activity with ferricyanide
693285
Sulfurisphaera tokodaii
pyruvate + reduced 2,6-dichlorophenolindophenol
-
-
-
?
(R)-lactate + oxidized 2,6-dichlorophenolindophenol
activity is 59% compared to the activity with ferricyanide
693285
Sulfurisphaera tokodaii 7
pyruvate + reduced 2,6-dichlorophenolindophenol
-
-
-
?
(R)-lactate + oxidized 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide
activity is 29% compared to the activity with ferricyanide
693285
Sulfurisphaera tokodaii
pyruvate + reduced 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide
-
-
-
?
(R)-lactate + oxidized 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide
activity is 29% compared to the activity with ferricyanide
693285
Sulfurisphaera tokodaii 7
pyruvate + reduced 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide
-
-
-
?
additional information
L-lactate, glycolate, L-tartrate, citrate, glyoxylate, 2-hydroxyisobutyrate, D-glycerate and D-2-phosphoglycerate are inert as electron donors. Benzyl viologen and methylene blue are inert as electron acceptors
693285
Sulfurisphaera tokodaii
?
-
-
-
-
additional information
L-lactate, glycolate, L-tartrate, citrate, glyoxylate, 2-hydroxyisobutyrate, D-glycerate and D-2-phosphoglycerate are inert as electron donors. Benzyl viologen and methylene blue are inert as electron acceptors
693285
Sulfurisphaera tokodaii 7
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
homodimer
2 * 48000, SDS-PAGE; 2 * 49715, calculated from sequence
Sulfurisphaera tokodaii
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
60
-
assay at
Sulfurisphaera tokodaii
80
-
-
Sulfurisphaera tokodaii
Temperature Stability [°C] (protein specific)
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
80
-
10 min, enzyme retains full activity
Sulfurisphaera tokodaii
85
-
10 min, more than 60% loss of activity
Sulfurisphaera tokodaii
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8.5
-
60°C
Sulfurisphaera tokodaii
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
6.5
9
pH 6.5: about 50% of maximal activity, pH 9.0: about 50% of maximal activity
Sulfurisphaera tokodaii
pH Stability (protein specific)
pH Stability
pH Stability Maximum
Commentary
Organism
6.5
10
50°C, 30 min, enzyme retains full activity
Sulfurisphaera tokodaii
Other publictions for EC 1.1.99.6
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
721200
Shibahara
Crystallization and preliminar ...
Aeropyrum pernix, Aeropyrum pernix DSM 11879
Acta Crystallogr. Sect. F
67
1425-1427
2011
-
-
1
1
-
-
-
-
-
-
-
-
-
4
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
693285
Satomura
A novel flavin adenine dinucle ...
Sulfurisphaera tokodaii, Sulfurisphaera tokodaii 7
J. Biosci. Bioeng.
106
16-21
2008
-
-
1
-
-
-
5
3
-
-
3
-
-
8
-
-
1
-
-
-
-
-
10
1
2
-
2
-
1
1
1
1
-
-
-
-
-
1
1
-
-
-
-
5
-
3
-
-
3
-
-
-
-
1
-
-
-
-
10
1
2
-
2
-
1
1
1
-
-
-
-
-
-
-
721463
Pagala
Cellular localization of D-lac ...
Archaeoglobus fulgidus
Archaea
1
95-104
2002
-
-
-
-
-
-
-
-
1
-
-
1
-
1
-
-
-
-
-
2
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
1
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
-
2
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
287843
Huang
Cloning and characterization o ...
Homo sapiens
Biochem. Biophys. Res. Commun.
268
298-301
2000
-
-
1
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
287842
Hekmat
-
Production of pyruvate from (R ...
Proteus vulgaris
Enzyme Microb. Technol.
24
471-479
1999
-
1
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
722504
Reed
The Archaeoglobus fulgidus D-l ...
Archaeoglobus fulgidus
J. Bacteriol.
181
7580-7587
1999
4
-
1
-
-
-
2
1
-
1
-
-
-
3
-
-
1
-
-
-
-
-
3
-
2
1
1
-
1
1
-
1
-
-
-
4
-
1
1
-
-
-
-
2
-
1
-
1
-
-
-
-
-
1
-
-
-
-
3
-
2
1
1
-
1
1
-
-
-
-
-
-
-
-
287841
Trautwein
The (2R)-hydroxycarboxylate-vi ...
Proteus vulgaris
Eur. J. Biochem.
222
1025-1032
1994
-
-
-
-
-
-
1
-
-
1
2
-
-
1
1
-
1
-
-
-
2
-
5
1
-
-
-
-
-
-
-
3
-
-
-
-
-
-
3
-
-
-
-
1
-
-
-
1
2
-
-
1
-
1
-
-
2
-
5
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
287840
Cammack
D-2-hydroxy acid dehydrogenase ...
Oryctolagus cuniculus
Methods Enzymol.
41
323-329
1975
4
-
-
-
-
1
7
2
1
-
1
2
-
2
-
-
1
-
-
1
1
2
14
-
1
-
-
-
2
-
1
5
-
-
-
4
-
-
5
-
-
1
-
7
-
2
1
-
1
2
-
-
-
1
-
1
1
2
14
-
1
-
-
-
2
-
1
-
-
-
-
-
-
-
287839
Tubbs
Dehydrogenation of D-lactate b ...
Oryctolagus cuniculus
Biochim. Biophys. Acta
34
290-291
1959
4
-
-
-
-
-
6
1
1
1
-
1
-
1
-
-
1
-
-
2
-
-
2
-
2
-
-
-
1
-
-
3
-
-
-
4
-
-
3
-
-
-
-
6
-
1
1
1
-
1
-
-
-
1
-
2
-
-
2
-
2
-
-
-
1
-
-
-
-
-
-
-
-
-