BRENDA - Enzyme Database show
show all sequences of 1.2.1.89

Improvement of thermostable aldehyde dehydrogenase by directed evolution for application in Synthetic Cascade Biomanufacturing

Steffler, F.; Guterl, J.K.; Sieber, V.; Enzyme Microb. Technol. 53, 307-314 (2013)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
; expressed in Escherichia coli BL21(DE3) cells
Thermoplasma acidophilum
Engineering
Amino acid exchange
Commentary
Organism
34M/Y399C/S405N
the mutations enhance enzyme activity with the cofactor NAD+ by a factor of 55.7
Thermoplasma acidophilum
F34L
the mutation enhances enzyme activity with the cofactor NAD+ by a factor of 3.2
Thermoplasma acidophilum
F34M/D372N/S405N
the mutations enhance enzyme activity with the cofactor NAD+ by a factor of 7.2
Thermoplasma acidophilum
F34M/D372N/Y399C/S405N
the mutations enhance enzyme activity with the cofactor NAD+ by a factor of 35.1
Thermoplasma acidophilum
F34M/S405N
enhances enzyme activity with the cofactor NAD+ by a factor of eight; the mutations enhance enzyme activity with the cofactor NAD+ by a factor of 8.5 while impacts on enzyme stability and substrate specificity are negligible
Thermoplasma acidophilum
F34M/W271R/S405N
the mutations enhance enzyme activity with the cofactor NAD+ by a factor of 22.6
Thermoplasma acidophilum
F34M/W271S/S405N
the mutations enhance enzyme activity with the cofactor NAD+ by a factor of 42.5
Thermoplasma acidophilum
F34M/W271S/Y399C/S405N
the mutations enhance enzyme activity with the cofactor NAD+ by a factor of 1.1
Thermoplasma acidophilum
F34M/Y399C/S405N
enhances enzyme activity with the cofactor NAD+ by a factor of eight and enhances solubilty
Thermoplasma acidophilum
S405C
the mutation enhances enzyme activity with the cofactor NAD+ by a factor of 6
Thermoplasma acidophilum
S405N
the mutation enhances enzyme activity with the cofactor NAD+ by a factor of 7.4
Thermoplasma acidophilum
W271S
the mutations enhance enzyme activity with the cofactor NAD+ by a factor of 17.1
Thermoplasma acidophilum
Y399C
enhances the protein solubility after recombinant expression in Escherichia coli 6fold; the mutation enhances the protein solubility after recombinant expression in Escherichia coli 6fold and enhances enzyme activity with the cofactor NAD+ by a factor of 6.4
Thermoplasma acidophilum
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.021
-
NADP+
wild-type, pH 7.0, 50°C
Thermoplasma acidophilum
0.031
-
NADP+
mutant F34M/Y399C/S405N, pH 7.0, 50°C
Thermoplasma acidophilum
0.036
-
NADP+
mutant F34M/S405N, pH 7.0, 50°C
Thermoplasma acidophilum
16.7
-
NAD+
mutant enzyme F34M/Y399C/S405N, at pH 7.0 and 50°C; mutant F34M/Y399C/S405N, pH 7.0, 50°C
Thermoplasma acidophilum
17.6
-
NAD+
mutant enzyme F34M/S405N, at pH 7.0 and 50°C; mutant F34M/S405N, pH 7.0, 50°C
Thermoplasma acidophilum
22.7
-
NAD+
wild type enzyme, at pH 7.0 and 50°C; wild-type, pH 7.0, 50°C
Thermoplasma acidophilum
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
55000
-
x * 55000, SDS-PAGE
Thermoplasma acidophilum
Organic Solvent Stability
Organic Solvent
Commentary
Organism
isobutanol
wild-type, 50% residual activity at 3.1%, mutant F34M/S405N at 3.6%, mutant F34M/Y399C/S405N at 3.5%, respectively
Thermoplasma acidophilum
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Thermoplasma acidophilum
Q9HK01
-
-
Purification (Commentary)
Commentary
Organism
HiTrap column chromatography
Thermoplasma acidophilum
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
D-glyceraldehyde + NAD+
NAD+ is a very poor cofactor for wild-type
736023
Thermoplasma acidophilum
D-glycerate + NADH + H+
-
-
-
?
D-glyceraldehyde + NADP+ + H2O
-
736023
Thermoplasma acidophilum
D-glycerate + NADPH + H+
-
-
-
?
glyceraldehyde + NAD+ + H2O
-
736023
Thermoplasma acidophilum
glycerate + NADH + H+
-
-
-
?
additional information
no activity with acetaldehyde and isobutyraldehyde
736023
Thermoplasma acidophilum
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
?
x * 55000, SDS-PAGE
Thermoplasma acidophilum
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.64
-
NAD+
wild type enzyme, at pH 7.0 and 50°C; wild-type, pH 7.0, 50°C
Thermoplasma acidophilum
10.54
-
NAD+
mutant enzyme F34M/Y399C/S405N, at pH 7.0 and 50°C; mutant F34M/Y399C/S405N, pH 7.0, 50°C
Thermoplasma acidophilum
10.99
-
NAD+
mutant enzyme F34M/S405N, at pH 7.0 and 50°C; mutant F34M/S405N, pH 7.0, 50°C
Thermoplasma acidophilum
Cofactor
Cofactor
Commentary
Organism
Structure
NAD+
-
Thermoplasma acidophilum
NADH
poor cofoactor
Thermoplasma acidophilum
NADPH
NADPH is preferred over NADH
Thermoplasma acidophilum
Cloned(Commentary) (protein specific)
Commentary
Organism
; expressed in Escherichia coli BL21(DE3) cells
Thermoplasma acidophilum
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NAD+
-
Thermoplasma acidophilum
NADH
poor cofoactor
Thermoplasma acidophilum
NADPH
NADPH is preferred over NADH
Thermoplasma acidophilum
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
34M/Y399C/S405N
the mutations enhance enzyme activity with the cofactor NAD+ by a factor of 55.7
Thermoplasma acidophilum
F34L
the mutation enhances enzyme activity with the cofactor NAD+ by a factor of 3.2
Thermoplasma acidophilum
F34M/D372N/S405N
the mutations enhance enzyme activity with the cofactor NAD+ by a factor of 7.2
Thermoplasma acidophilum
F34M/D372N/Y399C/S405N
the mutations enhance enzyme activity with the cofactor NAD+ by a factor of 35.1
Thermoplasma acidophilum
F34M/S405N
enhances enzyme activity with the cofactor NAD+ by a factor of eight; the mutations enhance enzyme activity with the cofactor NAD+ by a factor of 8.5 while impacts on enzyme stability and substrate specificity are negligible
Thermoplasma acidophilum
F34M/W271R/S405N
the mutations enhance enzyme activity with the cofactor NAD+ by a factor of 22.6
Thermoplasma acidophilum
F34M/W271S/S405N
the mutations enhance enzyme activity with the cofactor NAD+ by a factor of 42.5
Thermoplasma acidophilum
F34M/W271S/Y399C/S405N
the mutations enhance enzyme activity with the cofactor NAD+ by a factor of 1.1
Thermoplasma acidophilum
F34M/Y399C/S405N
enhances enzyme activity with the cofactor NAD+ by a factor of eight and enhances solubilty
Thermoplasma acidophilum
S405C
the mutation enhances enzyme activity with the cofactor NAD+ by a factor of 6
Thermoplasma acidophilum
S405N
the mutation enhances enzyme activity with the cofactor NAD+ by a factor of 7.4
Thermoplasma acidophilum
W271S
the mutations enhance enzyme activity with the cofactor NAD+ by a factor of 17.1
Thermoplasma acidophilum
Y399C
enhances the protein solubility after recombinant expression in Escherichia coli 6fold; the mutation enhances the protein solubility after recombinant expression in Escherichia coli 6fold and enhances enzyme activity with the cofactor NAD+ by a factor of 6.4
Thermoplasma acidophilum
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.021
-
NADP+
wild-type, pH 7.0, 50°C
Thermoplasma acidophilum
0.031
-
NADP+
mutant F34M/Y399C/S405N, pH 7.0, 50°C
Thermoplasma acidophilum
0.036
-
NADP+
mutant F34M/S405N, pH 7.0, 50°C
Thermoplasma acidophilum
16.7
-
NAD+
mutant enzyme F34M/Y399C/S405N, at pH 7.0 and 50°C; mutant F34M/Y399C/S405N, pH 7.0, 50°C
Thermoplasma acidophilum
17.6
-
NAD+
mutant enzyme F34M/S405N, at pH 7.0 and 50°C; mutant F34M/S405N, pH 7.0, 50°C
Thermoplasma acidophilum
22.7
-
NAD+
wild type enzyme, at pH 7.0 and 50°C; wild-type, pH 7.0, 50°C
Thermoplasma acidophilum
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
55000
-
x * 55000, SDS-PAGE
Thermoplasma acidophilum
Organic Solvent Stability (protein specific)
Organic Solvent
Commentary
Organism
isobutanol
wild-type, 50% residual activity at 3.1%, mutant F34M/S405N at 3.6%, mutant F34M/Y399C/S405N at 3.5%, respectively
Thermoplasma acidophilum
Purification (Commentary) (protein specific)
Commentary
Organism
HiTrap column chromatography
Thermoplasma acidophilum
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
D-glyceraldehyde + NAD+
NAD+ is a very poor cofactor for wild-type
736023
Thermoplasma acidophilum
D-glycerate + NADH + H+
-
-
-
?
D-glyceraldehyde + NADP+ + H2O
-
736023
Thermoplasma acidophilum
D-glycerate + NADPH + H+
-
-
-
?
glyceraldehyde + NAD+ + H2O
-
736023
Thermoplasma acidophilum
glycerate + NADH + H+
-
-
-
?
additional information
no activity with acetaldehyde and isobutyraldehyde
736023
Thermoplasma acidophilum
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
?
x * 55000, SDS-PAGE
Thermoplasma acidophilum
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.64
-
NAD+
wild type enzyme, at pH 7.0 and 50°C; wild-type, pH 7.0, 50°C
Thermoplasma acidophilum
10.54
-
NAD+
mutant enzyme F34M/Y399C/S405N, at pH 7.0 and 50°C; mutant F34M/Y399C/S405N, pH 7.0, 50°C
Thermoplasma acidophilum
10.99
-
NAD+
mutant enzyme F34M/S405N, at pH 7.0 and 50°C; mutant F34M/S405N, pH 7.0, 50°C
Thermoplasma acidophilum
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.07
-
NAD+
wild type enzyme, at pH 7.0 and 50°C; wild-type, pH 7.0, 50°C
Thermoplasma acidophilum
0.62
-
NAD+
mutant enzyme F34M/S405N, at pH 7.0 and 50°C; mutant F34M/S405N, pH 7.0, 50°C
Thermoplasma acidophilum
0.63
-
NAD+
mutant enzyme F34M/Y399C/S405N, at pH 7.0 and 50°C; mutant F34M/Y399C/S405N, pH 7.0, 50°C
Thermoplasma acidophilum
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.07
-
NAD+
wild type enzyme, at pH 7.0 and 50°C; wild-type, pH 7.0, 50°C
Thermoplasma acidophilum
0.62
-
NAD+
mutant enzyme F34M/S405N, at pH 7.0 and 50°C; mutant F34M/S405N, pH 7.0, 50°C
Thermoplasma acidophilum
0.63
-
NAD+
mutant enzyme F34M/Y399C/S405N, at pH 7.0 and 50°C; mutant F34M/Y399C/S405N, pH 7.0, 50°C
Thermoplasma acidophilum
Other publictions for EC 1.2.1.89
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)
740187
Wu
A new NAD(+)-dependent glycera ...
Escherichia coli
Biosci. Biotechnol. Biochem.
80
2306-2310
2016
-
-
-
-
3
-
-
3
-
-
-
-
-
2
-
-
-
-
-
-
3
-
1
-
-
-
1
3
-
-
-
1
-
-
-
-
-
-
1
-
3
-
-
-
-
3
-
-
-
-
-
-
-
-
-
-
3
-
1
-
-
-
1
3
-
-
-
-
-
-
-
-
3
3
739818
Iermak
-
Crystallization behaviour of g ...
Thermoplasma acidophilum
Acta Crystallogr. Sect. F
71
1475-1480
2015
-
-
1
1
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
726348
Steffler
Refolding of a thermostable gl ...
Thermoplasma acidophilum, Thermoplasma acidophilum JCM 9062
PLoS One
8
e70592
2013
-
1
1
-
-
-
-
1
-
-
-
-
2
6
-
-
1
-
-
-
-
-
4
-
1
-
-
-
1
-
-
1
-
-
-
-
1
1
1
-
-
-
-
-
-
1
-
-
-
-
2
-
-
1
-
-
-
-
4
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
736023
Steffler
Improvement of thermostable al ...
Thermoplasma acidophilum
Enzyme Microb. Technol.
53
307-314
2013
-
-
1
-
13
-
-
6
-
-
1
-
1
1
-
-
1
-
-
-
-
-
4
1
-
-
-
3
-
-
-
3
-
-
-
-
-
1
3
-
13
-
-
-
-
6
-
-
1
-
1
-
-
1
-
-
-
-
4
1
-
-
-
3
-
-
-
-
-
-
-
-
3
3
724216
Jung
Identification and characteriz ...
Thermoplasma acidophilum, Thermoplasma acidophilum JCM 9062
Biochem. J.
397
131-138
2006
5
-
-
-
-
-
8
2
-
-
3
2
-
8
-
-
1
-
-
-
1
-
10
2
1
1
2
-
1
1
1
1
-
-
-
5
-
-
1
-
-
-
-
8
-
2
-
-
3
2
-
-
-
1
-
-
1
-
10
2
1
1
2
-
1
1
1
-
-
1
1
-
-
-
724992
Reher
Glyceraldehyde dehydrogenases ...
Picrophilus torridus, Picrophilus torridus DSM 9790, Thermoplasma acidophilum, Thermoplasma acidophilum JCM 9062
FEBS Lett.
580
1198-1204
2006
-
-
1
-
-
-
1
8
-
-
6
4
-
9
-
-
2
-
-
-
1
-
18
2
4
-
4
8
4
2
-
2
-
-
-
-
-
1
2
-
-
-
-
1
-
8
-
-
6
4
-
-
-
2
-
-
1
-
18
2
4
-
4
8
4
2
-
-
-
2
2
-
8
8