BRENDA - Enzyme Database
show all sequences of 1.1.1.168

Recombinant expression and molecular insights into the catalytic mechanism of an NADPH-dependent conjugated polyketone reductase for the asymmetric synthesis of (R)-pantolactone

Cheng, P.; Wang, J.; Wu, Y.; Jiang, X.; Pei, X.; Su, W.; Enzyme Microb. Technol. 126, 77-85 (2019) View publication on PubMed

Data extracted from this reference:

Cloned(Commentary)
Cloned (Commentary)
Organism
expressed in Escherichia coli BL21(DE3) cells
Candida dubliniensis
Engineering
Protein Variants
Commentary
Organism
H128A
the mutant shows strongly reduced activity compared to the wild type enzyme
Candida dubliniensis
T30A
the mutant shows reduced activity compared to the wild type enzyme
Candida dubliniensis
Y66A
inactive
Candida dubliniensis
Inhibitors
Inhibitors
Commentary
Organism
Structure
Ag+
complete inhibition at 1 mM
Candida dubliniensis
Ca2+
94.6% residual activity at 1 mM
Candida dubliniensis
Co2+
75.3% residual activity at 1 mM
Candida dubliniensis
Cu2+
29.7% residual activity at 1 mM
Candida dubliniensis
EDTA
85.7% residual activity at 1 mM
Candida dubliniensis
Fe2+
66.8% residual activity at 1 mM
Candida dubliniensis
Mg2+
77.6% residual activity at 1 mM
Candida dubliniensis
Mn2+
56.8% residual activity at 1 mM
Candida dubliniensis
Ni2+
88.5% residual activity at 1 mM
Candida dubliniensis
Pb2+
complete inhibition at 1 mM
Candida dubliniensis
Zn2+
10.8% residual activity at 1 mM
Candida dubliniensis
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.484
-
2-dehydropantolactone
wild type enzyme, at pH 7.0 and 30°C
Candida dubliniensis
3.5
-
2-dehydropantolactone
mutant enzyme T30A, at pH 7.0 and 30°C
Candida dubliniensis
20.4
-
2-dehydropantolactone
mutant enzyme H128A, at pH 7.0 and 30°C
Candida dubliniensis
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
2-dehydropantolactone + NADPH + H+
Candida dubliniensis
98% conversion and 99% enantiomeric excess within 2 h
(R)-pantolactone + NADP+
-
-
?
2-dehydropantolactone + NADPH + H+
Candida dubliniensis CD36
98% conversion and 99% enantiomeric excess within 2 h
(R)-pantolactone + NADP+
-
-
?
Organism
Organism
UniProt
Commentary
Textmining
Candida dubliniensis
B9WJQ5
-
-
Candida dubliniensis CD36
B9WJQ5
-
-
Purification (Commentary)
Purification (Commentary)
Organism
Ni-NTA column chromatography
Candida dubliniensis
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
10.3
-
mutant enzyme H128A, at pH 7.0 and 30°C
Candida dubliniensis
26.5
-
mutant enzyme T30A, at pH 7.0 and 30°C
Candida dubliniensis
49.1
-
wild type enzyme, at pH 7.0 and 30°C
Candida dubliniensis
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
2-dehydropantolactone + NADPH + H+
98% conversion and 99% enantiomeric excess within 2 h
761008
Candida dubliniensis
(R)-pantolactone + NADP+
-
-
-
?
2-dehydropantolactone + NADPH + H+
98% conversion and 99% enantiomeric excess within 2 h
761008
Candida dubliniensis CD36
(R)-pantolactone + NADP+
-
-
-
?
Subunits
Subunits
Commentary
Organism
?
x * 37000, SDS-PAGE
Candida dubliniensis
Synonyms
Synonyms
Commentary
Organism
CPR
-
Candida dubliniensis
NADPH-dependent conjugated polyketone reductase
-
Candida dubliniensis
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
40
-
-
Candida dubliniensis
Temperature Range [°C]
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
35
55
between 35 and 55°C, the enzyme activity remains above 70% of the maximal activity
Candida dubliniensis
Temperature Stability [°C]
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
30
50
the residual activity of recombinant enzyme maintains above 90% of the initial activity after 7 h at 30°C. When the enzyme is incubated at 50°C, above 30% of the initial activity still retains after 2 h. The recombinant enzyme exhibits adequate thermal stability, and its half-lives are 45.6, 8.6 and 1.7 h at 30, 40 and 50°C, respectively
Candida dubliniensis
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
6.01
-
2-dehydropantolactone
mutant enzyme H128A, at pH 7.0 and 30°C
Candida dubliniensis
15.5
-
2-dehydropantolactone
mutant enzyme T30A, at pH 7.0 and 30°C
Candida dubliniensis
59
-
2-dehydropantolactone
wild type enzyme, at pH 7.0 and 30°C
Candida dubliniensis
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6.5
-
-
Candida dubliniensis
pH Stability
pH Stability
pH Stability Maximum
Commentary
Organism
6.5
7
the half-lives of the enzyme are 12.8 and 8.9 h at pH 6.5 and pH 7.0, respectively
Candida dubliniensis
Cofactor
Cofactor
Commentary
Organism
Structure
NADP+
-
Candida dubliniensis
Cloned(Commentary) (protein specific)
Commentary
Organism
expressed in Escherichia coli BL21(DE3) cells
Candida dubliniensis
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NADP+
-
Candida dubliniensis
Engineering (protein specific)
Protein Variants
Commentary
Organism
H128A
the mutant shows strongly reduced activity compared to the wild type enzyme
Candida dubliniensis
T30A
the mutant shows reduced activity compared to the wild type enzyme
Candida dubliniensis
Y66A
inactive
Candida dubliniensis
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
Ag+
complete inhibition at 1 mM
Candida dubliniensis
Ca2+
94.6% residual activity at 1 mM
Candida dubliniensis
Co2+
75.3% residual activity at 1 mM
Candida dubliniensis
Cu2+
29.7% residual activity at 1 mM
Candida dubliniensis
EDTA
85.7% residual activity at 1 mM
Candida dubliniensis
Fe2+
66.8% residual activity at 1 mM
Candida dubliniensis
Mg2+
77.6% residual activity at 1 mM
Candida dubliniensis
Mn2+
56.8% residual activity at 1 mM
Candida dubliniensis
Ni2+
88.5% residual activity at 1 mM
Candida dubliniensis
Pb2+
complete inhibition at 1 mM
Candida dubliniensis
Zn2+
10.8% residual activity at 1 mM
Candida dubliniensis
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.484
-
2-dehydropantolactone
wild type enzyme, at pH 7.0 and 30°C
Candida dubliniensis
3.5
-
2-dehydropantolactone
mutant enzyme T30A, at pH 7.0 and 30°C
Candida dubliniensis
20.4
-
2-dehydropantolactone
mutant enzyme H128A, at pH 7.0 and 30°C
Candida dubliniensis
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
2-dehydropantolactone + NADPH + H+
Candida dubliniensis
98% conversion and 99% enantiomeric excess within 2 h
(R)-pantolactone + NADP+
-
-
?
2-dehydropantolactone + NADPH + H+
Candida dubliniensis CD36
98% conversion and 99% enantiomeric excess within 2 h
(R)-pantolactone + NADP+
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
Ni-NTA column chromatography
Candida dubliniensis
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
10.3
-
mutant enzyme H128A, at pH 7.0 and 30°C
Candida dubliniensis
26.5
-
mutant enzyme T30A, at pH 7.0 and 30°C
Candida dubliniensis
49.1
-
wild type enzyme, at pH 7.0 and 30°C
Candida dubliniensis
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
2-dehydropantolactone + NADPH + H+
98% conversion and 99% enantiomeric excess within 2 h
761008
Candida dubliniensis
(R)-pantolactone + NADP+
-
-
-
?
2-dehydropantolactone + NADPH + H+
98% conversion and 99% enantiomeric excess within 2 h
761008
Candida dubliniensis CD36
(R)-pantolactone + NADP+
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
?
x * 37000, SDS-PAGE
Candida dubliniensis
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
40
-
-
Candida dubliniensis
Temperature Range [°C] (protein specific)
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
35
55
between 35 and 55°C, the enzyme activity remains above 70% of the maximal activity
Candida dubliniensis
Temperature Stability [°C] (protein specific)
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
30
50
the residual activity of recombinant enzyme maintains above 90% of the initial activity after 7 h at 30°C. When the enzyme is incubated at 50°C, above 30% of the initial activity still retains after 2 h. The recombinant enzyme exhibits adequate thermal stability, and its half-lives are 45.6, 8.6 and 1.7 h at 30, 40 and 50°C, respectively
Candida dubliniensis
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
6.01
-
2-dehydropantolactone
mutant enzyme H128A, at pH 7.0 and 30°C
Candida dubliniensis
15.5
-
2-dehydropantolactone
mutant enzyme T30A, at pH 7.0 and 30°C
Candida dubliniensis
59
-
2-dehydropantolactone
wild type enzyme, at pH 7.0 and 30°C
Candida dubliniensis
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6.5
-
-
Candida dubliniensis
pH Stability (protein specific)
pH Stability
pH Stability Maximum
Commentary
Organism
6.5
7
the half-lives of the enzyme are 12.8 and 8.9 h at pH 6.5 and pH 7.0, respectively
Candida dubliniensis
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.295
-
2-dehydropantolactone
mutant enzyme H128A, at pH 7.0 and 30°C
Candida dubliniensis
4.43
-
2-dehydropantolactone
mutant enzyme T30A, at pH 7.0 and 30°C
Candida dubliniensis
122
-
2-dehydropantolactone
wild type enzyme, at pH 7.0 and 30°C
Candida dubliniensis
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.295
-
2-dehydropantolactone
mutant enzyme H128A, at pH 7.0 and 30°C
Candida dubliniensis
4.43
-
2-dehydropantolactone
mutant enzyme T30A, at pH 7.0 and 30°C
Candida dubliniensis
122
-
2-dehydropantolactone
wild type enzyme, at pH 7.0 and 30°C
Candida dubliniensis
Other publictions for EC 1.1.1.168
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
Synonyms
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)
761008
Cheng
Recombinant expression and mo ...
Candida dubliniensis, Candida dubliniensis CD36
Enzyme Microb. Technol.
126
77-85
2019
-
-
1
-
3
-
11
3
-
-
-
2
-
12
-
-
1
-
-
-
3
-
2
1
2
1
1
1
3
1
-
1
1
-
-
-
-
-
1
1
-
3
-
-
11
-
3
-
-
-
2
-
-
-
1
-
-
3
-
2
1
1
1
1
3
1
-
1
-
-
-
-
-
3
3
761570
Wang
Discovery of a new NADPH-depe ...
Candida orthopsilosis, Candida orthopsilosis 90-125
J. Biotechnol.
291
26-34
2019
-
-
1
-
-
-
9
1
-
1
-
2
-
6
-
-
1
-
-
-
-
-
13
1
2
1
1
1
-
1
1
-
1
-
-
-
-
-
1
1
-
-
-
-
9
-
1
-
1
-
2
-
-
-
1
-
-
-
-
13
1
1
1
1
-
1
1
-
-
-
-
-
-
-
-
347731
Julliard
-
Purification and characterizat ...
Spinacia oleracea
Bot. Acta
107
191-200
1994
-
-
-
-
-
-
3
4
-
-
1
-
-
1
-
-
1
-
-
-
-
1
6
1
-
-
-
1
-
1
-
-
1
1
-
-
-
-
-
1
-
-
-
-
3
1
4
-
-
1
-
-
-
-
1
-
-
-
1
6
1
-
-
1
-
1
-
-
-
-
-
-
-
-
-
347730
Kataoka
Enzymes involved in the NADPH ...
Saccharomyces cerevisiae, Candida parapsilosis, Mucor ambiguus, Mucor ambiguus AKU 3006
Biosci. Biotechnol. Biochem.
56
820-821
1992
-
-
-
-
-
-
-
-
-
-
-
-
-
5
-
-
-
-
-
-
-
-
7
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
7
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
347723
Hata
Ketopantoyl-lactone reductase ...
Saccharomyces cerevisiae
FEMS Microbiol. Lett.
58
87-90
1989
-
-
-
-
-
-
3
16
-
-
-
1
-
4
-
-
-
-
-
-
-
-
21
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
4
-
16
-
-
-
1
-
-
-
-
-
-
-
-
21
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
347724
Hata
Ketopantoyl-lactone reductase ...
Candida parapsilosis
Biochim. Biophys. Acta
990
175-181
1989
-
-
-
1
-
-
11
4
-
-
2
2
-
10
-
-
1
-
-
-
2
-
12
1
-
1
-
2
-
-
-
2
-
-
-
-
-
-
-
-
1
-
-
-
12
-
4
-
-
2
2
-
-
-
1
-
-
2
-
12
1
1
-
2
-
-
-
2
-
-
-
-
-
-
-
347726
Wilken
Ketopantoyl lactone reductases ...
Saccharomyces cerevisiae
Methods Enzymol.
62
209-215
1979
-
-
-
-
-
1
5
-
-
-
1
1
-
1
-
-
1
-
-
-
1
1
4
-
-
1
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
1
-
5
-
-
-
-
1
1
-
-
-
1
-
-
1
1
4
-
1
-
-
-
2
-
-
-
-
-
-
-
-
-
347725
Wilken
Stereospecificity of pantoyl l ...
Saccharomyces cerevisiae
Arch. Biochem. Biophys.
189
251-255
1978
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
1
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
1
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
286105
Wilken
Ketopantoic acid and ketopanto ...
Saccharomyces cerevisiae
J. Biol. Chem.
250
2311-2314
1975
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
347727
King
Ketopantoyl lactone and ketopa ...
Saccharomyces cerevisiae
J. Biol. Chem.
249
4689-4695
1974
-
-
-
-
-
-
-
4
-
-
1
1
-
1
-
-
1
-
-
-
-
-
2
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
4
-
-
1
1
-
-
-
1
-
-
-
-
2
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
286104
King
-
Separation and preliminary stu ...
Saccharomyces cerevisiae
J. Biol. Chem.
247
4096-4098
1972
-
-
-
-
-
-
-
-
-
-
-
2
-
1
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
4
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-