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show all sequences of 1.1.1.B52

Gene cloning and characterization of two NADH-dependent 3-quinuclidinone reductases from Microbacterium luteolum JCM 9174

Isotani, K.; Kurokawa, J.; Suzuki, F.; Nomoto, S.; Negishi, T.; Matsuda, M.; Itoh, N.; Appl. Environ. Microbiol. 79, 1378-1384 (2012)

Data extracted from this reference:

Application
Application
Commentary
Organism
pharmacology
stereospecific production of (R)-3-quinuclidinol, an important chiral building block for the synthesis of various pharmaceuticals
Microbacterium luteolum
synthesis
stereospecific production of (R)-3-quinuclidinol, an important chiral building block for the synthesis of various pharmaceuticals
Microbacterium luteolum
Cloned(Commentary)
Commentary
Organism
expression in Escherichia coli. The gene is fused with the His6 tag and thrombin recognition sequence at the N-terminus under the control of the T7 promoter, 3-quinuclidinone reductase BacC; expression in Escherichia coli. The gene is fused with the His6 tag and thrombin recognition sequence at the N-terminus under the control of the T7 promoter, 3-quinuclidinone reductase QNR
Microbacterium luteolum
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.02
-
NADH
pH 7.0, 25°C, 3-quinuclidinone reductase QNR
Microbacterium luteolum
0.03
-
NADH
pH 7.0, 25°C, 3-quinuclidinone reductase BacC
Microbacterium luteolum
6.5
-
3-quinuclidinone
pH 7.0, 25°C, 3-quinuclidinone reductase QNR
Microbacterium luteolum
13.8
-
3-quinuclidinone
pH 7.0, 25°C, 3-quinuclidinone reductase BacC
Microbacterium luteolum
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
30600
-
3 * 30600, SDS-PAGE, 3-quinuclidinone reductase QNR
Microbacterium luteolum
32600
-
2 * 32600, SDS-PAGE, 3-quinuclidinone reductase BacC
Microbacterium luteolum
72000
-
gel filtration, 3-quinuclidinone reductase BacC
Microbacterium luteolum
94000
-
gel filtration, 3-quinuclidinone reductase QNR
Microbacterium luteolum
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Microbacterium luteolum
L8B2H6
-
-
Microbacterium luteolum JCM 9174
L8B2H6
-
-
Purification (Commentary)
Commentary
Organism
; 3-quinuclidinone reductase QNR
Microbacterium luteolum
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
0.5
-
pH 7.0, 25°C, 3-quinuclidinone reductase BacC
Microbacterium luteolum
8.4
-
pH 7.0, 25°C
Microbacterium luteolum
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2-acetylpyridine + NADH + H+
activity is 16.8% compared to the activity with 3-quinuclidinone, 3-quinuclidinone reductase BacC
729032
Microbacterium luteolum
?
-
-
-
?
2-acetylpyridine + NADH + H+
activity is 16.8% compared to the activity with 3-quinuclidinone, 3-quinuclidinone reductase BacC
729032
Microbacterium luteolum JCM 9174
?
-
-
-
?
3-quinuclidinone + NADH + H+
3-quinuclidinone (5% w/v, 313 mM) is reduced to (R)-3-quinuclidinol with a molar conversion yield of 100% by 3-quinuclidinone reductase QNR. The optical purity is above 99.9%. No activity with NADPH. The enzyme is strictly specific for 3-quinuclidinone and shows no activity toward several ketones, including tropinone
729032
Microbacterium luteolum
(R)-3-quinuclidinol + NAD+
-
-
-
?
3-quinuclidinone + NADH + H+
3-quinuclidinone (5% w/v, 313 mM) is reduced to (R)-3-quinuclidinol with a molar conversion yield of 94% bei 3-quinuclidinone reductase BacC. The optical purity is above 99.9%. No activity with NADPH
729032
Microbacterium luteolum
(R)-3-quinuclidinol + NAD+
-
-
-
?
3-quinuclidinone + NADH + H+
3-quinuclidinone (5% w/v, 313 mM) is reduced to (R)-3-quinuclidinol with a molar conversion yield of 100% by 3-quinuclidinone reductase QNR. The optical purity is above 99.9%. No activity with NADPH. The enzyme is strictly specific for 3-quinuclidinone and shows no activity toward several ketones, including tropinone
729032
Microbacterium luteolum JCM 9174
(R)-3-quinuclidinol + NAD+
-
-
-
?
3-quinuclidinone + NADH + H+
3-quinuclidinone (5% w/v, 313 mM) is reduced to (R)-3-quinuclidinol with a molar conversion yield of 94% bei 3-quinuclidinone reductase BacC. The optical purity is above 99.9%. No activity with NADPH
729032
Microbacterium luteolum JCM 9174
(R)-3-quinuclidinol + NAD+
-
-
-
?
7-oxabicyclo[4.1.0]heptan-2-one + NADH + H+
activity is 27.8% compared to the activity with 3-quinuclidinone, 3-quinuclidinone reductase BacC
729032
Microbacterium luteolum
?
-
-
-
?
7-oxabicyclo[4.1.0]heptan-2-one + NADH + H+
activity is 27.8% compared to the activity with 3-quinuclidinone, 3-quinuclidinone reductase BacC
729032
Microbacterium luteolum JCM 9174
?
-
-
-
?
Subunits
Subunits
Commentary
Organism
dimer
2 * 32600, SDS-PAGE, 3-quinuclidinone reductase BacC
Microbacterium luteolum
trimer
3 * 30600, SDS-PAGE, 3-quinuclidinone reductase QNR
Microbacterium luteolum
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
25
-
assay at
Microbacterium luteolum
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
8
3-quinuclidinone reductase QNR
Microbacterium luteolum
7
-
3-quinuclidinone reductase BacC
Microbacterium luteolum
Cofactor
Cofactor
Commentary
Organism
Structure
NADH
no activity with NADPH, 3-quinuclidinone reductase BacC; no activity with NADPH, 3-quinuclidinone reductase QNR
Microbacterium luteolum
Application (protein specific)
Application
Commentary
Organism
pharmacology
stereospecific production of (R)-3-quinuclidinol, an important chiral building block for the synthesis of various pharmaceuticals
Microbacterium luteolum
synthesis
stereospecific production of (R)-3-quinuclidinol, an important chiral building block for the synthesis of various pharmaceuticals
Microbacterium luteolum
Cloned(Commentary) (protein specific)
Commentary
Organism
expression in Escherichia coli. The gene is fused with the His6 tag and thrombin recognition sequence at the N-terminus under the control of the T7 promoter, 3-quinuclidinone reductase BacC; expression in Escherichia coli. The gene is fused with the His6 tag and thrombin recognition sequence at the N-terminus under the control of the T7 promoter, 3-quinuclidinone reductase QNR
Microbacterium luteolum
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NADH
no activity with NADPH, 3-quinuclidinone reductase BacC; no activity with NADPH, 3-quinuclidinone reductase QNR
Microbacterium luteolum
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.02
-
NADH
pH 7.0, 25°C, 3-quinuclidinone reductase QNR
Microbacterium luteolum
0.03
-
NADH
pH 7.0, 25°C, 3-quinuclidinone reductase BacC
Microbacterium luteolum
6.5
-
3-quinuclidinone
pH 7.0, 25°C, 3-quinuclidinone reductase QNR
Microbacterium luteolum
13.8
-
3-quinuclidinone
pH 7.0, 25°C, 3-quinuclidinone reductase BacC
Microbacterium luteolum
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
30600
-
3 * 30600, SDS-PAGE, 3-quinuclidinone reductase QNR
Microbacterium luteolum
32600
-
2 * 32600, SDS-PAGE, 3-quinuclidinone reductase BacC
Microbacterium luteolum
72000
-
gel filtration, 3-quinuclidinone reductase BacC
Microbacterium luteolum
94000
-
gel filtration, 3-quinuclidinone reductase QNR
Microbacterium luteolum
Purification (Commentary) (protein specific)
Commentary
Organism
; 3-quinuclidinone reductase QNR
Microbacterium luteolum
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
0.5
-
pH 7.0, 25°C, 3-quinuclidinone reductase BacC
Microbacterium luteolum
8.4
-
pH 7.0, 25°C
Microbacterium luteolum
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2-acetylpyridine + NADH + H+
activity is 16.8% compared to the activity with 3-quinuclidinone, 3-quinuclidinone reductase BacC
729032
Microbacterium luteolum
?
-
-
-
?
2-acetylpyridine + NADH + H+
activity is 16.8% compared to the activity with 3-quinuclidinone, 3-quinuclidinone reductase BacC
729032
Microbacterium luteolum JCM 9174
?
-
-
-
?
3-quinuclidinone + NADH + H+
3-quinuclidinone (5% w/v, 313 mM) is reduced to (R)-3-quinuclidinol with a molar conversion yield of 100% by 3-quinuclidinone reductase QNR. The optical purity is above 99.9%. No activity with NADPH. The enzyme is strictly specific for 3-quinuclidinone and shows no activity toward several ketones, including tropinone
729032
Microbacterium luteolum
(R)-3-quinuclidinol + NAD+
-
-
-
?
3-quinuclidinone + NADH + H+
3-quinuclidinone (5% w/v, 313 mM) is reduced to (R)-3-quinuclidinol with a molar conversion yield of 94% bei 3-quinuclidinone reductase BacC. The optical purity is above 99.9%. No activity with NADPH
729032
Microbacterium luteolum
(R)-3-quinuclidinol + NAD+
-
-
-
?
3-quinuclidinone + NADH + H+
3-quinuclidinone (5% w/v, 313 mM) is reduced to (R)-3-quinuclidinol with a molar conversion yield of 100% by 3-quinuclidinone reductase QNR. The optical purity is above 99.9%. No activity with NADPH. The enzyme is strictly specific for 3-quinuclidinone and shows no activity toward several ketones, including tropinone
729032
Microbacterium luteolum JCM 9174
(R)-3-quinuclidinol + NAD+
-
-
-
?
3-quinuclidinone + NADH + H+
3-quinuclidinone (5% w/v, 313 mM) is reduced to (R)-3-quinuclidinol with a molar conversion yield of 94% bei 3-quinuclidinone reductase BacC. The optical purity is above 99.9%. No activity with NADPH
729032
Microbacterium luteolum JCM 9174
(R)-3-quinuclidinol + NAD+
-
-
-
?
7-oxabicyclo[4.1.0]heptan-2-one + NADH + H+
activity is 27.8% compared to the activity with 3-quinuclidinone, 3-quinuclidinone reductase BacC
729032
Microbacterium luteolum
?
-
-
-
?
7-oxabicyclo[4.1.0]heptan-2-one + NADH + H+
activity is 27.8% compared to the activity with 3-quinuclidinone, 3-quinuclidinone reductase BacC
729032
Microbacterium luteolum JCM 9174
?
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
dimer
2 * 32600, SDS-PAGE, 3-quinuclidinone reductase BacC
Microbacterium luteolum
trimer
3 * 30600, SDS-PAGE, 3-quinuclidinone reductase QNR
Microbacterium luteolum
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
25
-
assay at
Microbacterium luteolum
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
8
3-quinuclidinone reductase QNR
Microbacterium luteolum
7
-
3-quinuclidinone reductase BacC
Microbacterium luteolum
Other publictions for EC 1.1.1.B52
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)
729146
Hou
Structural basis for high subs ...
Agrobacterium tumefaciens
Biochem. Biophys. Res. Commun.
446
911-915
2014
-
2
1
1
4
1
-
-
-
-
-
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1
-
-
1
-
-
-
-
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1
-
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-
1
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1
-
-
-
-
2
1
1
1
4
1
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
738052
Spickermann
Engineering of highly selectiv ...
Parvibaculum lavamentivorans
ChemBioChem
15
2050-2052
2014
-
-
1
-
11
-
-
-
-
-
-
-
-
1
-
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-
-
-
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2
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1
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11
-
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-
2
-
-
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-
-
-
-
-
-
-
-
-
-
-
-
730545
Zhang
Highly efficient synthesis of ...
Agrobacterium tumefaciens, Agrobacterium tumefaciens ECU2556
Org. Lett.
15
4917-4919
2013
-
2
1
-
-
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1
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2
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2
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2
1
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1
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-
2
-
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-
-
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-
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-
-
-
1
1
728893
Hou
Expression, purification, crys ...
Agrobacterium tumefaciens
Acta Crystallogr. Sect. F
68
1237-1239
2012
-
-
1
1
-
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
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-
1
-
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1
1
1
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-
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1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
729032
Isotani
Gene cloning and characterizat ...
Microbacterium luteolum, Microbacterium luteolum JCM 9174
Appl. Environ. Microbiol.
79
1378-1384
2012
-
2
1
-
-
-
-
4
-
-
4
-
-
3
-
-
1
-
-
-
2
-
8
2
1
-
-
-
2
-
-
1
-
-
-
-
2
1
1
-
-
-
-
-
-
4
-
-
4
-
-
-
-
1
-
-
2
-
8
2
1
-
-
-
2
-
-
-
-
-
-
-
-
-
729854
Isotani
Production of (R)-3-quinuclidi ...
Microbacterium luteolum, Microbacterium luteolum JCM 9174
Int. J. Mol. Sci.
13
13542-13553
2012
-
2
-
-
-
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
2
-
1
-
-
-
1
-
-
1
-
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-
-
2
-
1
-
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-
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2
-
1
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1
-
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