BRENDA - Enzyme Database show
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 (2013)

Data extracted from this reference:

Application
Application
Commentary
Organism
synthesis
(R)-3-quinuclidinol is a valuable intermediate for pharmaceuticals. The enzyme can be used for the synthesis of the enantiopure compound
Microbacterium luteolum
Cloned(Commentary)
Commentary
Organism
gene bacC, library construction, DNA and amino acid sequence determination and analysis, recombinant expression of His-tagged enzyme in Escherichia coli; gene qnr, library construction, DNA and amino acid sequence determination and analysis, recombinant expression of His-tagged enzyme in Escherichia coli
Microbacterium luteolum
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.02
-
NADH
recombinant enzyme, pH 7.0, 25°C
Microbacterium luteolum
0.03
-
NADH
recombinant enzyme, pH 7.0, 25°C
Microbacterium luteolum
6.5
-
3-quinuclidinone
recombinant enzyme, pH 7.0, 25°C
Microbacterium luteolum
13.8
-
3-quinuclidinone
recombinant enzyme, pH 7.0, 25°C
Microbacterium luteolum
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
3-quinuclidinone + NADH + H+
Microbacterium luteolum
-
(R)-3-quinuclidinol + NAD+
-
-
ir
3-quinuclidinone + NADH + H+
Microbacterium luteolum JCM 9174
-
(R)-3-quinuclidinol + NAD+
-
-
ir
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Microbacterium luteolum
L8B2H6
i.e. Aureobacterium luteolum
-
Microbacterium luteolum JCM 9174
L8B2H6
i.e. Aureobacterium luteolum
-
Purification (Commentary)
Commentary
Organism
native enzyme by anion exchange chromatography, dialysis, hydrophobic interaction chromatography, and dialysis, recombinant His-tagged enzyme from Escherichia coli 1.7fold by nickel affinity chromatography and ultrafiltration; native enzyme by anion exchange chromatography, dialysis, hydrophobic interaction chromatography, and dialysis, recombinant His-tagged enzyme from Escherichia coli 2.4fold by nickel affinity chromatography and ultrafiltration
Microbacterium luteolum
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2-acetylpyridine + NADH + H+
16.8% of the activity with 3-quinuclidinone
741676
Microbacterium luteolum
? + NAD+
-
-
-
?
2-acetylpyridine + NADH + H+
16.8% of the activity with 3-quinuclidinone
741676
Microbacterium luteolum JCM 9174
? + NAD+
-
-
-
?
3-quinuclidinone + NADH + H+
-
741676
Microbacterium luteolum
(R)-3-quinuclidinol + NAD+
-
-
-
ir
3-quinuclidinone + NADH + H+
over 99.9% enantiomeric excess
741676
Microbacterium luteolum
(R)-3-quinuclidinol + NAD+
-
-
-
ir
3-quinuclidinone + NADH + H+
-
741676
Microbacterium luteolum JCM 9174
(R)-3-quinuclidinol + NAD+
-
-
-
ir
3-quinuclidinone + NADH + H+
over 99.9% enantiomeric excess
741676
Microbacterium luteolum JCM 9174
(R)-3-quinuclidinol + NAD+
-
-
-
ir
7-oxabicyclo[4.1.0]heptan-2-one + NADH + H+
27.8% of the activity with 3-quinuclidinone
741676
Microbacterium luteolum
? + NAD+
-
-
-
?
additional information
the enzyme shows narrow substrate specificity and high stereoselectivity for the reduction of 3-quinuclidinone to (R)-3-quinuclidinol. No activity with (R)-3-quinuclidinol, 2-acetylpyridine, and 7-oxabicyclo[4.1.0]heptan-2-one, as well as with 2-propanol, acetone, acetophenone, 2,2,2-trifluoroacetophenone, 3-methylene-2-norbornanone, ethyl pyruvate, ethyl acetoacetate, 4-hydroxy-1-cyclohexanecarboxylic acid delta-lactone, verbenone, tropinone, 6-hydroxytropinone, 2-azabicyclo[2,2,1]hept-5-en-3-one, 1-methyl-4-piperidone, 3-acetylpyridine, 4-acetylpyridine, cyclohexanone, 2-methylcyclohexanone, 3-methylcyclohexanone, and tetrahydrothiopyran-4-one
741676
Microbacterium luteolum
?
-
-
-
-
additional information
the enzyme shows narrow substrate specificity and high stereoselectivity for the reduction of 3-quinuclidinone to (R)-3-quinuclidinol. No activity with (R)-3-quinuclidinol, 2-propanol, acetone, acetophenone, 2,2,2-trifluoroacetophenone, 3-methylene-2-norbornanone, ethyl pyruvate, ethyl acetoacetate, 4-hydroxy-1-cyclohexanecarboxylic acid delta-lactone, verbenone, tropinone, 6-hydroxytropinone, 2-azabicyclo[2,2,1]hept-5-en-3-one, 1-methyl-4-piperidone, 3-acetylpyridine, 4-acetylpyridine, cyclohexanone, 2-methylcyclohexanone, 3-methylcyclohexanone, and tetrahydrothiopyran-4-one
741676
Microbacterium luteolum
?
-
-
-
-
additional information
the enzyme shows narrow substrate specificity and high stereoselectivity for the reduction of 3-quinuclidinone to (R)-3-quinuclidinol. No activity with (R)-3-quinuclidinol, 2-acetylpyridine, and 7-oxabicyclo[4.1.0]heptan-2-one, as well as with 2-propanol, acetone, acetophenone, 2,2,2-trifluoroacetophenone, 3-methylene-2-norbornanone, ethyl pyruvate, ethyl acetoacetate, 4-hydroxy-1-cyclohexanecarboxylic acid delta-lactone, verbenone, tropinone, 6-hydroxytropinone, 2-azabicyclo[2,2,1]hept-5-en-3-one, 1-methyl-4-piperidone, 3-acetylpyridine, 4-acetylpyridine, cyclohexanone, 2-methylcyclohexanone, 3-methylcyclohexanone, and tetrahydrothiopyran-4-one
741676
Microbacterium luteolum JCM 9174
?
-
-
-
-
additional information
the enzyme shows narrow substrate specificity and high stereoselectivity for the reduction of 3-quinuclidinone to (R)-3-quinuclidinol. No activity with (R)-3-quinuclidinol, 2-propanol, acetone, acetophenone, 2,2,2-trifluoroacetophenone, 3-methylene-2-norbornanone, ethyl pyruvate, ethyl acetoacetate, 4-hydroxy-1-cyclohexanecarboxylic acid delta-lactone, verbenone, tropinone, 6-hydroxytropinone, 2-azabicyclo[2,2,1]hept-5-en-3-one, 1-methyl-4-piperidone, 3-acetylpyridine, 4-acetylpyridine, cyclohexanone, 2-methylcyclohexanone, 3-methylcyclohexanone, and tetrahydrothiopyran-4-one
741676
Microbacterium luteolum JCM 9174
?
-
-
-
-
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
-
Microbacterium luteolum
7
-
-
Microbacterium luteolum
pH Range
pH Minimum
pH Maximum
Commentary
Organism
5.5
8.5
activity range, profile overview
Microbacterium luteolum
5.5
9.5
activity range, profile overview
Microbacterium luteolum
Cofactor
Cofactor
Commentary
Organism
Structure
NADH
-
Microbacterium luteolum
Application (protein specific)
Application
Commentary
Organism
synthesis
(R)-3-quinuclidinol is a valuable intermediate for pharmaceuticals. The enzyme can be used for the synthesis of the enantiopure compound
Microbacterium luteolum
Cloned(Commentary) (protein specific)
Commentary
Organism
gene bacC, library construction, DNA and amino acid sequence determination and analysis, recombinant expression of His-tagged enzyme in Escherichia coli; gene qnr, library construction, DNA and amino acid sequence determination and analysis, recombinant expression of His-tagged enzyme in Escherichia coli
Microbacterium luteolum
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NADH
-
Microbacterium luteolum
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.02
-
NADH
recombinant enzyme, pH 7.0, 25°C
Microbacterium luteolum
0.03
-
NADH
recombinant enzyme, pH 7.0, 25°C
Microbacterium luteolum
6.5
-
3-quinuclidinone
recombinant enzyme, pH 7.0, 25°C
Microbacterium luteolum
13.8
-
3-quinuclidinone
recombinant enzyme, pH 7.0, 25°C
Microbacterium luteolum
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
3-quinuclidinone + NADH + H+
Microbacterium luteolum
-
(R)-3-quinuclidinol + NAD+
-
-
ir
3-quinuclidinone + NADH + H+
Microbacterium luteolum JCM 9174
-
(R)-3-quinuclidinol + NAD+
-
-
ir
Purification (Commentary) (protein specific)
Commentary
Organism
native enzyme by anion exchange chromatography, dialysis, hydrophobic interaction chromatography, and dialysis, recombinant His-tagged enzyme from Escherichia coli 1.7fold by nickel affinity chromatography and ultrafiltration; native enzyme by anion exchange chromatography, dialysis, hydrophobic interaction chromatography, and dialysis, recombinant His-tagged enzyme from Escherichia coli 2.4fold by nickel affinity chromatography and ultrafiltration
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+
16.8% of the activity with 3-quinuclidinone
741676
Microbacterium luteolum
? + NAD+
-
-
-
?
2-acetylpyridine + NADH + H+
16.8% of the activity with 3-quinuclidinone
741676
Microbacterium luteolum JCM 9174
? + NAD+
-
-
-
?
3-quinuclidinone + NADH + H+
-
741676
Microbacterium luteolum
(R)-3-quinuclidinol + NAD+
-
-
-
ir
3-quinuclidinone + NADH + H+
over 99.9% enantiomeric excess
741676
Microbacterium luteolum
(R)-3-quinuclidinol + NAD+
-
-
-
ir
3-quinuclidinone + NADH + H+
-
741676
Microbacterium luteolum JCM 9174
(R)-3-quinuclidinol + NAD+
-
-
-
ir
3-quinuclidinone + NADH + H+
over 99.9% enantiomeric excess
741676
Microbacterium luteolum JCM 9174
(R)-3-quinuclidinol + NAD+
-
-
-
ir
7-oxabicyclo[4.1.0]heptan-2-one + NADH + H+
27.8% of the activity with 3-quinuclidinone
741676
Microbacterium luteolum
? + NAD+
-
-
-
?
additional information
the enzyme shows narrow substrate specificity and high stereoselectivity for the reduction of 3-quinuclidinone to (R)-3-quinuclidinol. No activity with (R)-3-quinuclidinol, 2-acetylpyridine, and 7-oxabicyclo[4.1.0]heptan-2-one, as well as with 2-propanol, acetone, acetophenone, 2,2,2-trifluoroacetophenone, 3-methylene-2-norbornanone, ethyl pyruvate, ethyl acetoacetate, 4-hydroxy-1-cyclohexanecarboxylic acid delta-lactone, verbenone, tropinone, 6-hydroxytropinone, 2-azabicyclo[2,2,1]hept-5-en-3-one, 1-methyl-4-piperidone, 3-acetylpyridine, 4-acetylpyridine, cyclohexanone, 2-methylcyclohexanone, 3-methylcyclohexanone, and tetrahydrothiopyran-4-one
741676
Microbacterium luteolum
?
-
-
-
-
additional information
the enzyme shows narrow substrate specificity and high stereoselectivity for the reduction of 3-quinuclidinone to (R)-3-quinuclidinol. No activity with (R)-3-quinuclidinol, 2-propanol, acetone, acetophenone, 2,2,2-trifluoroacetophenone, 3-methylene-2-norbornanone, ethyl pyruvate, ethyl acetoacetate, 4-hydroxy-1-cyclohexanecarboxylic acid delta-lactone, verbenone, tropinone, 6-hydroxytropinone, 2-azabicyclo[2,2,1]hept-5-en-3-one, 1-methyl-4-piperidone, 3-acetylpyridine, 4-acetylpyridine, cyclohexanone, 2-methylcyclohexanone, 3-methylcyclohexanone, and tetrahydrothiopyran-4-one
741676
Microbacterium luteolum
?
-
-
-
-
additional information
the enzyme shows narrow substrate specificity and high stereoselectivity for the reduction of 3-quinuclidinone to (R)-3-quinuclidinol. No activity with (R)-3-quinuclidinol, 2-acetylpyridine, and 7-oxabicyclo[4.1.0]heptan-2-one, as well as with 2-propanol, acetone, acetophenone, 2,2,2-trifluoroacetophenone, 3-methylene-2-norbornanone, ethyl pyruvate, ethyl acetoacetate, 4-hydroxy-1-cyclohexanecarboxylic acid delta-lactone, verbenone, tropinone, 6-hydroxytropinone, 2-azabicyclo[2,2,1]hept-5-en-3-one, 1-methyl-4-piperidone, 3-acetylpyridine, 4-acetylpyridine, cyclohexanone, 2-methylcyclohexanone, 3-methylcyclohexanone, and tetrahydrothiopyran-4-one
741676
Microbacterium luteolum JCM 9174
?
-
-
-
-
additional information
the enzyme shows narrow substrate specificity and high stereoselectivity for the reduction of 3-quinuclidinone to (R)-3-quinuclidinol. No activity with (R)-3-quinuclidinol, 2-propanol, acetone, acetophenone, 2,2,2-trifluoroacetophenone, 3-methylene-2-norbornanone, ethyl pyruvate, ethyl acetoacetate, 4-hydroxy-1-cyclohexanecarboxylic acid delta-lactone, verbenone, tropinone, 6-hydroxytropinone, 2-azabicyclo[2,2,1]hept-5-en-3-one, 1-methyl-4-piperidone, 3-acetylpyridine, 4-acetylpyridine, cyclohexanone, 2-methylcyclohexanone, 3-methylcyclohexanone, and tetrahydrothiopyran-4-one
741676
Microbacterium luteolum JCM 9174
?
-
-
-
-
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
-
Microbacterium luteolum
7
-
-
Microbacterium luteolum
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
5.5
8.5
activity range, profile overview
Microbacterium luteolum
5.5
9.5
activity range, profile overview
Microbacterium luteolum
General Information
General Information
Commentary
Organism
evolution
the enzyme belongs to the short-chain alcohol dehydrogenase/reductase (SDR) family
Microbacterium luteolum
General Information (protein specific)
General Information
Commentary
Organism
evolution
the enzyme belongs to the short-chain alcohol dehydrogenase/reductase (SDR) family
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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3
-
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1
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1
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1
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1
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2
1
1
1
4
1
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1
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1
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1
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-
-
-
-
-
-
738052
Spickermann
Engineering of highly selectiv ...
Parvibaculum lavamentivorans
ChemBioChem
15
2050-2052
2014
-
-
1
-
11
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
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2
-
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1
-
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11
-
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-
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-
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-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
741816
Hou
Structural basis for high sub ...
Agrobacterium tumefaciens
Biochem. Biophys. Res. Commun.
446
911-915
2014
-
-
1
1
4
-
-
-
-
-
-
1
-
3
-
-
1
-
-
-
-
-
1
1
1
-
-
-
1
-
-
1
-
-
-
-
-
1
1
1
4
-
-
-
-
-
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-
-
1
-
-
-
1
-
-
-
-
1
1
1
-
-
-
1
-
-
-
-
2
2
-
-
-
730545
Zhang
Highly efficient synthesis of ...
Agrobacterium tumefaciens
Org. Lett.
15
4917-4919
2013
-
2
1
-
-
-
-
1
-
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1
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1
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2
1
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-
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1
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-
-
-
-
-
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-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
1
1
741676
Isotani
Gene cloning and characteriza ...
Microbacterium luteolum, Microbacterium luteolum JCM 9174
Appl. Environ. Microbiol.
79
1378-1384
2013
-
1
1
-
-
-
-
4
-
-
-
2
-
4
-
-
1
-
-
-
-
-
11
-
1
-
-
-
2
2
-
1
-
-
-
-
1
1
1
-
-
-
-
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4
-
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-
2
-
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1
-
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-
-
11
-
1
-
-
-
2
2
-
-
-
1
1
-
-
-
728893
Hou
Expression, purification, crys ...
Agrobacterium tumefaciens
Acta Crystallogr. Sect. F
68
1237-1239
2012
-
-
1
1
-
-
-
-
-
-
-
-
-
3
-
-
1
-
-
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1
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1
1
1
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1
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729032
Isotani
Gene cloning and characterizat ...
Microbacterium luteolum, Microbacterium luteolum JCM 9174
Appl. Environ. Microbiol.
79
1378-1384
2012
-
2
1
-
-
-
-
4
-
-
4
-
-
4
-
-
1
-
-
-
2
-
8
2
1
-
-
-
2
-
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1
-
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2
1
1
-
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4
-
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4
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1
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2
-
8
2
1
-
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2
-
-
-
-
-
-
-
-
-
729854
Isotani
Production of (R)-3-quinuclidi ...
Microbacterium luteolum, Microbacterium luteolum JCM 9174
Int. J. Mol. Sci.
13
13542-13553
2012
-
2
-
-
-
-
-
-
-
-
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9
-
-
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-
-
-
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2
-
1
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1
-
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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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