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

Cloning, expression, and directed evolution of carbonyl reductase from Leifsonia xyli HS0904 with enhanced catalytic efficiency

Wang, N.Q.; Sun, J.; Huang, J.; Wang, P.; Appl. Microbiol. Biotechnol. 98, 8591-8601 (2014)

Data extracted from this reference:

Application
Application
Commentary
Organism
synthesis
using recombinant Escherichia coli cells expressing Sdr, a yield of 82.5% for (R)-[3,5-bis(trifluoromethyl)phenyl]ethanol can be achieved within 12 h at a substrate concentration of up to 1000 M
Leifsonia xyli
Engineering
Amino acid exchange
Commentary
Organism
S154Y
mutant exhibits nearly 13fold, 5.4fold, and 2.3fold increase in kcat/Km value, kcat value, and specific activity toward 3,5-bis(trifluoromethyl)acetophenone
Leifsonia xyli
S154Y/L194I
180% of wild-type activity
Leifsonia xyli
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.025
-
NADH
wild-type, pH 7.2, 30°C
Leifsonia xyli
0.026
-
NADH
mutant S154Y/L194I, pH 7.2, 30°C
Leifsonia xyli
0.029
-
NADH
mutant S154Y, pH 7.2, 30°C
Leifsonia xyli
0.2
-
3,5-bis(trifluoromethyl)acetophenone
mutant S154Y, pH 7.2, 30°C
Leifsonia xyli
0.32
-
3,5-bis(trifluoromethyl)acetophenone
mutant S154Y/L194I, pH 7.2, 30°C
Leifsonia xyli
0.57
-
3,5-bis(trifluoromethyl)acetophenone
wild-type, pH 7.2, 30°C
Leifsonia xyli
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Leifsonia xyli
T2FLN4
-
-
Leifsonia xyli HS0904
T2FLN4
-
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3,5-bis(trifluoromethyl)acetophenone + NADH + H+
-
739942
Leifsonia xyli
(R)-[3,5-bis(trifluoromethyl)phenyl]ethanol + NAD+
-
-
-
?
3,5-bis(trifluoromethyl)acetophenone + NADH + H+
-
739942
Leifsonia xyli HS0904
(R)-[3,5-bis(trifluoromethyl)phenyl]ethanol + NAD+
-
-
-
?
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
21.1
-
3,5-bis(trifluoromethyl)acetophenone
wild-type, pH 7.2, 30°C
Leifsonia xyli
23.6
-
NADH
wild-type, pH 7.2, 30°C
Leifsonia xyli
26.6
-
NADH
mutant S154Y, pH 7.2, 30°C
Leifsonia xyli
89.2
-
3,5-bis(trifluoromethyl)acetophenone
mutant S154Y/L194I, pH 7.2, 30°C
Leifsonia xyli
100
-
NADH
mutant S154Y/L194I, pH 7.2, 30°C
Leifsonia xyli
114
-
3,5-bis(trifluoromethyl)acetophenone
mutant S154Y, pH 7.2, 30°C
Leifsonia xyli
Application (protein specific)
Application
Commentary
Organism
synthesis
using recombinant Escherichia coli cells expressing Sdr, a yield of 82.5% for (R)-[3,5-bis(trifluoromethyl)phenyl]ethanol can be achieved within 12 h at a substrate concentration of up to 1000 M
Leifsonia xyli
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
S154Y
mutant exhibits nearly 13fold, 5.4fold, and 2.3fold increase in kcat/Km value, kcat value, and specific activity toward 3,5-bis(trifluoromethyl)acetophenone
Leifsonia xyli
S154Y/L194I
180% of wild-type activity
Leifsonia xyli
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.025
-
NADH
wild-type, pH 7.2, 30°C
Leifsonia xyli
0.026
-
NADH
mutant S154Y/L194I, pH 7.2, 30°C
Leifsonia xyli
0.029
-
NADH
mutant S154Y, pH 7.2, 30°C
Leifsonia xyli
0.2
-
3,5-bis(trifluoromethyl)acetophenone
mutant S154Y, pH 7.2, 30°C
Leifsonia xyli
0.32
-
3,5-bis(trifluoromethyl)acetophenone
mutant S154Y/L194I, pH 7.2, 30°C
Leifsonia xyli
0.57
-
3,5-bis(trifluoromethyl)acetophenone
wild-type, pH 7.2, 30°C
Leifsonia xyli
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3,5-bis(trifluoromethyl)acetophenone + NADH + H+
-
739942
Leifsonia xyli
(R)-[3,5-bis(trifluoromethyl)phenyl]ethanol + NAD+
-
-
-
?
3,5-bis(trifluoromethyl)acetophenone + NADH + H+
-
739942
Leifsonia xyli HS0904
(R)-[3,5-bis(trifluoromethyl)phenyl]ethanol + NAD+
-
-
-
?
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
21.1
-
3,5-bis(trifluoromethyl)acetophenone
wild-type, pH 7.2, 30°C
Leifsonia xyli
23.6
-
NADH
wild-type, pH 7.2, 30°C
Leifsonia xyli
26.6
-
NADH
mutant S154Y, pH 7.2, 30°C
Leifsonia xyli
89.2
-
3,5-bis(trifluoromethyl)acetophenone
mutant S154Y/L194I, pH 7.2, 30°C
Leifsonia xyli
100
-
NADH
mutant S154Y/L194I, pH 7.2, 30°C
Leifsonia xyli
114
-
3,5-bis(trifluoromethyl)acetophenone
mutant S154Y, pH 7.2, 30°C
Leifsonia xyli
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
36.8
-
3,5-bis(trifluoromethyl)acetophenone
wild-type, pH 7.2, 30°C
Leifsonia xyli
277
-
3,5-bis(trifluoromethyl)acetophenone
mutant S154Y/L194I, pH 7.2, 30°C
Leifsonia xyli
463
-
3,5-bis(trifluoromethyl)acetophenone
mutant S154Y, pH 7.2, 30°C
Leifsonia xyli
944
-
NADH
wild-type, pH 7.2, 30°C
Leifsonia xyli
3846
-
NADH
mutant S154Y/L194I, pH 7.2, 30°C
Leifsonia xyli
4207
-
NADH
mutant S154Y, pH 7.2, 30°C
Leifsonia xyli
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
36.8
-
3,5-bis(trifluoromethyl)acetophenone
wild-type, pH 7.2, 30°C
Leifsonia xyli
277
-
3,5-bis(trifluoromethyl)acetophenone
mutant S154Y/L194I, pH 7.2, 30°C
Leifsonia xyli
463
-
3,5-bis(trifluoromethyl)acetophenone
mutant S154Y, pH 7.2, 30°C
Leifsonia xyli
944
-
NADH
wild-type, pH 7.2, 30°C
Leifsonia xyli
3846
-
NADH
mutant S154Y/L194I, pH 7.2, 30°C
Leifsonia xyli
4207
-
NADH
mutant S154Y, pH 7.2, 30°C
Leifsonia xyli
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)
741784
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3
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1
1
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1
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17
1
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24
1
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17
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26
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742305
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16
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4
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1
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1
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740394
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3
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13
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1
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2
2
739942
Wang
Cloning, expression, and direc ...
Leifsonia xyli, Leifsonia xyli HS0904
Appl. Microbiol. Biotechnol.
98
8591-8601
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1
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6
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6
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6
6
743703
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Kocuria rhizophila
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1
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9
5
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1
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19
1
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3
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9
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1
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19
1
1
1
3
-
1
1
1
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721911
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1
1
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15
1
1
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1
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Aggarwal
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Candida parapsilosis, Candida parapsilosis ATCC 7330
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313-315
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Protsko
Crystallization and preliminar ...
Thermoanaerobacter ethanolicus
Acta Crystallogr. Sect. F
66
831-833
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1
1
1
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1
1
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3
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1
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1
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1
1
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1
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1
1
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1
1
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725726
Li
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Deracemization of aryl seconda ...
Microbacterium oxydans, Microbacterium oxydans ECU2010
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64
48-52
2010
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4
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Pal
Activity of yeast alcohol dehy ...
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3
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697839
Pennacchio
Role of Tryptophan 95 in subst ...
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Cofactor engineering of Lactob ...
Lactobacillus brevis
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Pennacchio
Purification and characterizat ...
Thermus thermophilus
Appl. Environ. Microbiol.
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1
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5
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4
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1
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7
1
2
1
4
4
2
2
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3
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1
1
1
3
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7
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4
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1
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7
1
2
1
4
4
2
2
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Nie
A novel NADH-dependent carbony ...
Candida parapsilosis
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Isobe
Thermostable NAD+-dependent (R ...
Burkholderia sp., Burkholderia sp. AIU 652
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24
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6
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24
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1
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1
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3
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1
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688571
Bradshaw
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A Pseudomonas sp. alcohol dehy ...
Pseudomonas sp., Pseudomonas sp. PED
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