BRENDA - Enzyme Database show
show all sequences of 1.1.1.71

Mutant alcohol dehydrogenase leads to improved ethanol tolerance in Clostridium thermocellum

Brown, S.D.; Guss, A.M.; Karpinets, T.V.; Parks, J.M.; Smolin, N.; Yang, S.; Land, M.L.; Klingeman, D.M.; Bhandiwad, A.; Rodriguez, M. Jr.; Raman, B.; Shao, X.; Mielenz, J.R.; Smith, J.C.; Keller, M.; Lynd, L.R.; Proc. Natl. Acad. Sci. USA 108, 13752-71375 (2011)

Data extracted from this reference:

Engineering
Amino acid exchange
Commentary
Organism
P704L/H734R
the ethanol tolerant phenotype of Clostridium thermocellum is primarily due to a mutated bifunctional acetaldehyde-CoA/alcohol dehydrogenase gene (adhE). The mutant displays a complete loss of NADH-dependent activity with concomitant acquisition of NADPH-dependent activity. The reduction in specific activity with respect to NADH is far greater (about 25fold less activity) than the increase with respect to NADPH. Although total ADH activity dropps by 25fold, ethanol production does not drop significantly between strains under these conditions
Ruminiclostridium thermocellum
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Ruminiclostridium thermocellum
A0A0H3W5U9
bifunctional enzyme, C-terminal section belongs to the iron-containing alcohol dehydrogenase family, N-terminal section belongs to the aldehyde dehydrogenase family
-
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
0.025
-
wild-type, cofoactor NADPH, pH 7.6, 55°C
Ruminiclostridium thermocellum
0.052
-
mutant P704L/H734R, cofoactor NADH, pH 7.6, 55°C
Ruminiclostridium thermocellum
0.12
-
mutant P704L/H734R, cofoactor NADPH, pH 7.6, 55°C
Ruminiclostridium thermocellum
2.7
-
wild-type, cofoactor NADH, pH 7.6, 55°C
Ruminiclostridium thermocellum
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
acetaldehyde + NADH + H+
-
739526
Ruminiclostridium thermocellum
ethanol + NAD+
-
-
-
?
acetaldehyde + NADPH + H+
-
739526
Ruminiclostridium thermocellum
ethanol + NADP+
-
-
-
?
Cofactor
Cofactor
Commentary
Organism
Structure
NADH
-
Ruminiclostridium thermocellum
NADPH
the ethanol tolerant phenotype of Clostridium thermocellum is primarily due to a mutated bifunctional acetaldehyde-CoA/alcohol dehydrogenase gene (adhE). The mutant displays a complete loss of NADH-dependent activity with concomitant acquisition of NADPH-dependent activity
Ruminiclostridium thermocellum
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NADH
-
Ruminiclostridium thermocellum
NADPH
the ethanol tolerant phenotype of Clostridium thermocellum is primarily due to a mutated bifunctional acetaldehyde-CoA/alcohol dehydrogenase gene (adhE). The mutant displays a complete loss of NADH-dependent activity with concomitant acquisition of NADPH-dependent activity
Ruminiclostridium thermocellum
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
P704L/H734R
the ethanol tolerant phenotype of Clostridium thermocellum is primarily due to a mutated bifunctional acetaldehyde-CoA/alcohol dehydrogenase gene (adhE). The mutant displays a complete loss of NADH-dependent activity with concomitant acquisition of NADPH-dependent activity. The reduction in specific activity with respect to NADH is far greater (about 25fold less activity) than the increase with respect to NADPH. Although total ADH activity dropps by 25fold, ethanol production does not drop significantly between strains under these conditions
Ruminiclostridium thermocellum
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
0.025
-
wild-type, cofoactor NADPH, pH 7.6, 55°C
Ruminiclostridium thermocellum
0.052
-
mutant P704L/H734R, cofoactor NADH, pH 7.6, 55°C
Ruminiclostridium thermocellum
0.12
-
mutant P704L/H734R, cofoactor NADPH, pH 7.6, 55°C
Ruminiclostridium thermocellum
2.7
-
wild-type, cofoactor NADH, pH 7.6, 55°C
Ruminiclostridium thermocellum
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
acetaldehyde + NADH + H+
-
739526
Ruminiclostridium thermocellum
ethanol + NAD+
-
-
-
?
acetaldehyde + NADPH + H+
-
739526
Ruminiclostridium thermocellum
ethanol + NADP+
-
-
-
?
Other publictions for EC 1.1.1.71
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)
738540
Zheng
Cofactor specificity of the bi ...
Ruminiclostridium thermocellum, Thermoanaerobacterium saccharolyticum
J. Bacteriol.
197
2610-2619
2015
-
-
-
-
-
-
-
5
-
-
-
-
-
2
-
-
-
-
-
-
-
-
6
-
-
-
-
6
-
-
-
3
-
-
-
-
-
-
3
-
-
-
-
-
-
5
-
-
-
-
-
-
-
-
-
-
-
-
6
-
-
-
-
6
-
-
-
-
-
-
-
-
4
4
726699
Stekhanova
Sodium chloride-induced modula ...
Thermococcus sibiricus, Thermococcus sibiricus DSM 12597
Appl. Biochem. Biotechnol.
171
1877-1889
2013
-
-
1
-
-
-
-
6
-
1
-
-
-
4
-
-
-
-
-
-
-
-
8
1
3
2
5
6
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
6
-
1
-
-
-
-
-
-
-
-
-
-
8
1
3
2
5
6
1
-
-
-
-
-
-
-
6
6
728288
Liliensiek
Heterologous overexpression, p ...
Halobacterium salinarum, Halobacterium sp., Halobacterium sp. NRC-1
Mol. Biotechnol.
55
143-149
2013
-
-
2
-
-
-
-
10
-
3
1
-
8
3
-
-
1
-
-
-
-
2
21
1
1
-
-
-
4
-
-
4
-
-
-
-
-
2
4
-
-
-
-
-
-
10
-
3
1
-
8
-
-
1
-
-
-
2
21
1
1
-
-
-
4
-
-
-
-
-
-
-
-
-
722208
Timpson
Characterization of alcohol de ...
Haloarcula marismortui
Extremophiles
16
57-66
2012
-
-
1
-
-
-
-
3
-
1
1
-
3
4
-
-
1
-
-
-
-
1
8
1
1
1
-
-
2
-
-
2
-
-
-
-
-
1
2
-
-
-
-
-
-
3
-
1
1
-
3
-
-
1
-
-
-
1
8
1
1
1
-
-
2
-
-
-
-
-
-
-
-
-
726769
Timpson
A comparison of two novel alco ...
Haloferax volcanii, Haloferax volcanii DSM 3757
Appl. Microbiol. Biotechnol.
97
195-203
2012
-
-
1
-
-
1
-
5
-
1
2
-
4
2
-
-
1
-
-
-
-
2
14
1
1
1
-
-
2
-
1
2
-
-
-
-
-
1
2
-
-
1
-
-
-
5
-
1
2
-
4
-
-
1
-
-
-
2
14
1
1
1
-
-
2
-
1
-
-
-
-
-
5
5
739526
Brown
Mutant alcohol dehydrogenase l ...
Ruminiclostridium thermocellum
Proc. Natl. Acad. Sci. USA
108
13752-71375
2011
-
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
4
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
2
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
4
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
726606
Lyashenko
Expression, purification and c ...
Thermococcus sibiricus, Thermococcus sibiricus DSM 12597
Acta Crystallogr. Sect. F
66
655-657
2010
-
-
1
1
-
-
-
-
-
-
-
-
-
4
-
-
1
-
-
-
-
-
4
-
1
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
4
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
726712
Stekhanova
Characterization of a thermost ...
Thermococcus sibiricus, Thermococcus sibiricus DSM 12597
Appl. Environ. Microbiol.
76
4096-4098
2010
-
-
1
-
-
-
14
5
-
3
3
-
-
7
-
-
1
-
-
-
-
-
30
1
1
-
4
5
2
-
-
1
-
-
-
-
-
1
1
-
-
-
-
14
-
5
-
3
3
-
-
-
-
1
-
-
-
-
30
1
1
-
4
5
2
-
-
-
-
-
-
-
5
5
668112
Thomas
Retinoid metabolism in the rat ...
Rattus norvegicus
Br. J. Nutr.
93
59-63
2005
-
-
-
-
-
-
-
-
1
-
-
1
-
2
-
-
-
-
-
3
-
-
2
-
1
-
-
-
1
-
-
2
-
-
-
-
-
-
2
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
-
3
-
-
2
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
657019
Montesano
A novel potato defence-related ...
Solanum tuberosum
Plant Mol. Biol.
52
177-189
2003
2
-
1
-
-
-
-
5
-
-
-
1
-
1
-
-
1
1
-
1
-
-
15
-
-
-
-
-
-
-
-
2
-
-
-
2
-
1
2
-
-
-
-
-
-
5
-
-
-
1
-
-
-
1
-
1
-
-
15
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
655563
Belyaeva
Human pancreas protein 2 (PAN2 ...
Homo sapiens
FEBS Lett.
531
489-493
2002
-
-
1
-
-
-
-
6
2
-
1
-
-
1
-
-
-
1
-
14
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
6
2
-
1
-
-
-
-
-
-
14
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
287273
Hirschberg
N-terminal acetylation in a th ...
Gallus gallus
Cell. Mol. Life Sci.
58
1323-1326
2001
-
-
-
-
-
-
-
-
-
-
1
-
-
3
-
-
-
1
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
287275
Korkhin
NADP-dependent bacterial alcoh ...
Clostridium beijerinckii, Thermoanaerobacter brockii
J. Mol. Biol.
278
967-981
1998
-
-
-
2
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
2
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
287277
Wales
NADP-dependent alcohol dehydro ...
Acinetobacter calcoaceticus, Bacillus subtilis, Bacillus subtilis NCIMB 3610, Escherichia coli, Escherichia coli ML30, Mycobacterium tuberculosis, Pseudomonas aeruginosa, Pseudomonas putida, Pseudomonas putida NCIMB 9494, Rhodococcus rhodochrous, Rhodococcus rhodochrous NCIMB 13259, Rhodotorula graminis, Rhodotorula graminis KGX 39, Saccharomyces cerevisiae, Saccharomyces cerevisiae D273-10B, Streptomyces rimosus 4018, Streptomyces rimosus
Microbiology
140
173-183
1994
-
-
-
-
-
-
6
17
-
-
2
-
-
20
-
-
2
2
-
-
-
-
35
2
-
-
-
13
4
2
-
10
-
-
-
-
-
-
10
-
-
-
-
6
-
17
-
-
2
-
-
-
-
2
-
-
-
-
35
2
-
-
-
13
4
2
-
-
-
-
-
-
-
-
287278
Brophy
A novel NADPH/NADH-dependent a ...
Moniezia expansa
FEBS Lett.
263
305-307
1990
-
-
-
-
-
-
2
2
1
-
-
-
-
1
-
-
1
-
-
-
-
-
8
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
1
-
-
-
-
2
-
2
1
-
-
-
-
-
-
1
-
-
-
-
8
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
287279
Hiu
Butanol-ethanol dehydrogenase ...
Clostridium beijerinckii
Appl. Environ. Microbiol.
53
697-703
1987
-
-
-
-
-
-
-
2
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
-
-
-
-
-
2
2
-
1
-
-
-
-
-
-
1
-
-
-
-
-
-
2
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
2
2
-
-
-
-
-
-
-
-
287280
Hatanaka
-
Purification and properties of ...
Leuconostoc mesenteroides, Leuconostoc mesenteroides IFO 3426
Agric. Biol. Chem.
38
1819-1833
1974
-
-
-
-
-
-
9
8
-
-
-
-
-
2
-
-
1
-
-
-
-
-
14
-
-
-
1
-
2
-
1
1
-
-
-
-
-
-
1
-
-
-
-
9
-
8
-
-
-
-
-
-
-
1
-
-
-
-
14
-
-
-
1
-
2
-
1
-
-
-
-
-
-
-
287281
Rhodes
-
Co-factor specificity of plant ...
Cucumis melo
Phytochemistry
12
307-314
1973
-
-
-
-
-
-
2
4
-
-
-
-
-
1
-
-
1
-
-
-
-
-
1
-
-
-
-
-
2
2
-
1
-
-
-
-
-
-
1
-
-
-
-
2
-
4
-
-
-
-
-
-
-
1
-
-
-
-
1
-
-
-
-
-
2
2
-
-
-
-
-
-
-
-
287282
Fidge
The enzymatic reduction of ret ...
Rattus norvegicus
J. Biol. Chem.
243
4372-4379
1968
2
-
-
-
-
-
5
-
-
-
-
-
-
1
-
-
1
-
-
-
-
-
14
-
-
-
-
-
1
1
-
1
-
-
-
2
-
-
1
-
-
-
-
5
-
-
-
-
-
-
-
-
-
1
-
-
-
-
14
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-