BRENDA - Enzyme Database show
show all sequences of 1.1.1.B4

Role of Tryptophan 95 in substrate specificity and structural stability of Sulfolobus solfataricus alcohol dehydrogenase

Pennacchio, A.; Esposito, L.; Zagari, A.; Rossi, M.; Raia, C.A.; Extremophiles 13, 751-761 (2009)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expressed in Escherichia coli RB791 cells
Sulfolobus solfataricus
Crystallization (Commentary)
Crystallization
Organism
-
Sulfolobus solfataricus
Engineering
Amino acid exchange
Commentary
Organism
W95L/N249Y
the mutant exhibits higher activity but decreased affinity toward aliphatic alcohols, aldehydes as well as NAD+ and NADH compared to the wild type enzyme, optimum pH is at about pH 8.6
Sulfolobus solfataricus
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.22
-
(R)-2-pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65C
Sulfolobus solfataricus
0.41
-
(R)-2-butanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65C
Sulfolobus solfataricus
11.7
-
(R)-2-pentanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65C
Sulfolobus solfataricus
37.4
-
(R)-2-butanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65C
Sulfolobus solfataricus
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Sulfolobus solfataricus
-
-
-
Purification (Commentary)
Commentary
Organism
-
Sulfolobus solfataricus
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(R)-2-butanol + NAD+
-
697839
Sulfolobus solfataricus
2-butanone + NADH + H+
-
-
-
r
(R)-2-pentanol + NAD+
-
697839
Sulfolobus solfataricus
2-pentanone + NADH + H+
-
-
-
r
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1
-
(R)-2-butanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65C
Sulfolobus solfataricus
1.1
-
(R)-2-pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65C
Sulfolobus solfataricus
1.7
-
(R)-2-butanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65C
Sulfolobus solfataricus
1.9
-
(R)-2-pentanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65C
Sulfolobus solfataricus
Cofactor
Cofactor
Commentary
Organism
Structure
NAD+
-
Sulfolobus solfataricus
Cloned(Commentary) (protein specific)
Commentary
Organism
expressed in Escherichia coli RB791 cells
Sulfolobus solfataricus
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NAD+
-
Sulfolobus solfataricus
Crystallization (Commentary) (protein specific)
Crystallization
Organism
-
Sulfolobus solfataricus
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
W95L/N249Y
the mutant exhibits higher activity but decreased affinity toward aliphatic alcohols, aldehydes as well as NAD+ and NADH compared to the wild type enzyme, optimum pH is at about pH 8.6
Sulfolobus solfataricus
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.22
-
(R)-2-pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65C
Sulfolobus solfataricus
0.41
-
(R)-2-butanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65C
Sulfolobus solfataricus
11.7
-
(R)-2-pentanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65C
Sulfolobus solfataricus
37.4
-
(R)-2-butanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65C
Sulfolobus solfataricus
Purification (Commentary) (protein specific)
Commentary
Organism
-
Sulfolobus solfataricus
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(R)-2-butanol + NAD+
-
697839
Sulfolobus solfataricus
2-butanone + NADH + H+
-
-
-
r
(R)-2-pentanol + NAD+
-
697839
Sulfolobus solfataricus
2-pentanone + NADH + H+
-
-
-
r
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1
-
(R)-2-butanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65C
Sulfolobus solfataricus
1.1
-
(R)-2-pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65C
Sulfolobus solfataricus
1.7
-
(R)-2-butanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65C
Sulfolobus solfataricus
1.9
-
(R)-2-pentanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65C
Sulfolobus solfataricus
Other publictions for EC 1.1.1.B4
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)
721911
Zhou
Biocatalytic characterization ...
Carboxydothermus hydrogenoformans, Carboxydothermus hydrogenoformans DSM 6008
Biotechnol. Lett.
35
359-365
2013
-
1
1
-
-
-
-
1
-
-
1
-
4
2
-
-
-
-
-
-
-
-
15
1
1
-
-
1
1
-
-
2
-
-
-
-
1
1
2
-
-
-
-
-
-
1
-
-
1
-
4
-
-
-
-
-
-
-
15
1
1
-
-
1
1
-
-
-
-
-
-
-
-
-
723841
Protsko
Crystallization and preliminar ...
Thermoanaerobacter ethanolicus
Acta Crystallogr. Sect. F
66
831-833
2010
-
-
1
1
1
-
-
-
-
-
1
1
-
1
-
-
1
-
-
-
-
-
1
1
-
-
-
-
-
-
-
2
-
-
-
-
-
1
2
1
1
-
-
-
-
-
-
-
1
1
-
-
-
1
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
1
1
-
-
-
725726
Li
-
Deracemization of aryl seconda ...
Microbacterium oxydans, Microbacterium oxydans ECU2010
J. Mol. Catal. B
64
48-52
2010
-
1
-
-
-
-
-
-
-
-
-
2
-
2
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
2
-
-
-
-
1
-
2
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
697267
Pal
Activity of yeast alcohol dehy ...
Saccharomyces carlsbergensis, Saccharomyces carlsbergensis Y379-50, Saccharomyces cerevisiae
Chem. Biol. Interact.
178
16-23
2009
-
-
-
-
-
-
-
-
1
-
-
-
-
3
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
2
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
697839
Pennacchio
Role of Tryptophan 95 in subst ...
Sulfolobus solfataricus
Extremophiles
13
751-761
2009
-
-
1
1
1
-
-
4
-
-
-
-
-
1
-
-
1
-
-
-
-
-
2
-
-
-
-
4
-
-
-
1
-
-
-
-
-
1
1
1
1
-
-
-
-
4
-
-
-
-
-
-
-
1
-
-
-
-
2
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
711860
Machielsen
-
Cofactor engineering of Lactob ...
Lactobacillus brevis
Eng. Life Sci.
9
38-44
2009
-
-
1
-
5
-
-
4
-
-
-
-
-
1
-
-
-
-
-
-
-
-
2
-
-
-
2
-
2
-
-
1
-
-
-
-
-
1
1
-
5
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
2
-
2
-
-
-
-
-
-
-
4
4
684583
Pennacchio
Purification and characterizat ...
Thermus thermophilus
Appl. Environ. Microbiol.
74
3949-3958
2008
1
1
1
-
-
-
7
5
-
5
4
-
1
1
-
-
1
-
-
-
-
-
7
1
2
1
4
4
2
2
-
3
-
-
-
1
1
1
3
-
-
-
-
7
-
5
-
5
4
-
1
-
-
1
-
-
-
-
7
1
2
1
4
4
2
2
-
-
-
-
-
-
-
-
688747
Nie
A novel NADH-dependent carbony ...
Candida parapsilosis
Lett. Appl. Microbiol.
44
555-562
2007
-
-
-
-
-
-
9
-
1
1
2
-
-
1
-
-
1
-
-
-
1
1
10
1
1
-
-
-
1
-
-
2
-
-
-
-
-
-
2
-
-
-
-
9
-
-
1
1
2
-
-
-
-
1
-
-
1
1
10
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
687864
Isobe
Thermostable NAD+-dependent (R ...
Burkholderia sp., Burkholderia sp. AIU 652
J. Biosci. Bioeng.
96
387-393
2003
-
-
-
-
-
-
6
15
-
-
2
-
-
3
-
-
1
-
-
-
1
-
24
1
1
-
1
-
3
-
-
3
-
1
-
-
-
-
3
-
-
-
-
6
-
15
-
-
2
-
-
-
-
1
-
-
1
-
24
1
1
-
1
-
3
-
-
1
-
-
-
-
-
-
688571
Bradshaw
-
A Pseudomonas sp. alcohol dehy ...
Pseudomonas sp., Pseudomonas sp. PED
J. Org. Chem.
57
1526-1532
1992
-
-
-
-
-
-
-
2
-
-
-
-
-
2
-
-
-
1
-
-
1
-
24
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
1
-
24
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-