BRENDA - Enzyme Database show
show all sequences of 1.1.1.346

Cloning, expression and characterization of a putative 2,5-diketo-D-gluconic acid reductase in Comamonas testosteroni

Chen, Y.; Ji, W.; Zhang, H.; Zhang, X.; Yu, Y.; Chem. Biol. Interact. 234, 229-235 (2015)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
gene CTATCC11996_22452, genetic organization, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis and tree, recombinant overexpression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
Comamonas testosteroni
Engineering
Amino acid exchange
Commentary
Organism
additional information
construction of enzyme gene knockout mutant M-AKR, that shows decreased degradation activity with testosterone, estradiol, oestrone, and methyltestosterone compared to the wild-type enzyme. Compared to the wild-type, the mutation of the endogenous 2,5DKR gene results in lower degradation of estradiol and methyltestosterone but has no effct on degradation of estrone and testosterone
Comamonas testosteroni
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2-dehydro-L-gulonate + NADP+
Comamonas testosteroni
-
2,5-didehydro-D-gluconate + NADPH + H+
-
-
?
2-dehydro-L-gulonate + NADP+
Comamonas testosteroni ATCC 11996
-
2,5-didehydro-D-gluconate + NADPH + H+
-
-
?
additional information
Comamonas testosteroni
the enzyme catalyzes degradation of estradiol, oestrone, testosterone, and methyltestosterone, overview
?
-
-
-
additional information
Comamonas testosteroni ATCC 11996
the enzyme catalyzes degradation of estradiol, oestrone, testosterone, and methyltestosterone, overview
?
-
-
-
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Comamonas testosteroni
H1RW56
-
-
Comamonas testosteroni ATCC 11996
H1RW56
-
-
Purification (Commentary)
Commentary
Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
Comamonas testosteroni
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2-dehydro-L-gulonate + NADP+
-
742259
Comamonas testosteroni
2,5-didehydro-D-gluconate + NADPH + H+
-
-
-
?
2-dehydro-L-gulonate + NADP+
-
742259
Comamonas testosteroni ATCC 11996
2,5-didehydro-D-gluconate + NADPH + H+
-
-
-
?
additional information
the enzyme catalyzes degradation of estradiol, oestrone, testosterone, and methyltestosterone, overview
742259
Comamonas testosteroni
?
-
-
-
-
additional information
the enzyme catalyzes degradation of estradiol, oestrone, testosterone, and methyltestosterone, overview
742259
Comamonas testosteroni ATCC 11996
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
?
x * 39400, about, sequence calculation, x * 41600, recombinant His-tagged enzyme, SDS-PAGE
Comamonas testosteroni
Cofactor
Cofactor
Commentary
Organism
Structure
NADP+
-
Comamonas testosteroni
Cloned(Commentary) (protein specific)
Commentary
Organism
gene CTATCC11996_22452, genetic organization, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis and tree, recombinant overexpression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
Comamonas testosteroni
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NADP+
-
Comamonas testosteroni
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
additional information
construction of enzyme gene knockout mutant M-AKR, that shows decreased degradation activity with testosterone, estradiol, oestrone, and methyltestosterone compared to the wild-type enzyme. Compared to the wild-type, the mutation of the endogenous 2,5DKR gene results in lower degradation of estradiol and methyltestosterone but has no effct on degradation of estrone and testosterone
Comamonas testosteroni
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2-dehydro-L-gulonate + NADP+
Comamonas testosteroni
-
2,5-didehydro-D-gluconate + NADPH + H+
-
-
?
2-dehydro-L-gulonate + NADP+
Comamonas testosteroni ATCC 11996
-
2,5-didehydro-D-gluconate + NADPH + H+
-
-
?
additional information
Comamonas testosteroni
the enzyme catalyzes degradation of estradiol, oestrone, testosterone, and methyltestosterone, overview
?
-
-
-
additional information
Comamonas testosteroni ATCC 11996
the enzyme catalyzes degradation of estradiol, oestrone, testosterone, and methyltestosterone, overview
?
-
-
-
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
Comamonas testosteroni
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2-dehydro-L-gulonate + NADP+
-
742259
Comamonas testosteroni
2,5-didehydro-D-gluconate + NADPH + H+
-
-
-
?
2-dehydro-L-gulonate + NADP+
-
742259
Comamonas testosteroni ATCC 11996
2,5-didehydro-D-gluconate + NADPH + H+
-
-
-
?
additional information
the enzyme catalyzes degradation of estradiol, oestrone, testosterone, and methyltestosterone, overview
742259
Comamonas testosteroni
?
-
-
-
-
additional information
the enzyme catalyzes degradation of estradiol, oestrone, testosterone, and methyltestosterone, overview
742259
Comamonas testosteroni ATCC 11996
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
?
x * 39400, about, sequence calculation, x * 41600, recombinant His-tagged enzyme, SDS-PAGE
Comamonas testosteroni
Expression
Organism
Commentary
Expression
Comamonas testosteroni
the enzyme expression is significantly induced by estrone, estradiol, and methyltestosterone, and slightly by progesterone, but not by testosterone
up
General Information
General Information
Commentary
Organism
evolution
the enzyme belongs to the AKR superfamily, monomeric (alpha/beta) 8-barrel proteins which bind NAD(P)(H) to metabolize an array of substrates
Comamonas testosteroni
malfunction
compared to the wild-type, the knockout mutation of the endogenous 2,5DKR gene results in lower degradation of estradiol and methyltestosterone but has no effct on degradation of estrone and testosterone. Cell growth on ethanol, oestrone, estradiol, testosterone or methyltestosterone is reduced in the mutant strain compared to the wild-type
Comamonas testosteroni
additional information
three consensus sequences of the AKR superfamily are found as GxxxxDxAxxY, LxxxGxxxPxxGxG and LxxxxxxxxxDxxxxH. GxxxxDxAxxY is the active site, LxxxGxxxPxxGxG is the cofactor-binding site for NAD(P)(H), and LxxxxxxxxxDxxxxH is required for supporting the 3D structure
Comamonas testosteroni
General Information (protein specific)
General Information
Commentary
Organism
evolution
the enzyme belongs to the AKR superfamily, monomeric (alpha/beta) 8-barrel proteins which bind NAD(P)(H) to metabolize an array of substrates
Comamonas testosteroni
malfunction
compared to the wild-type, the knockout mutation of the endogenous 2,5DKR gene results in lower degradation of estradiol and methyltestosterone but has no effct on degradation of estrone and testosterone. Cell growth on ethanol, oestrone, estradiol, testosterone or methyltestosterone is reduced in the mutant strain compared to the wild-type
Comamonas testosteroni
additional information
three consensus sequences of the AKR superfamily are found as GxxxxDxAxxY, LxxxGxxxPxxGxG and LxxxxxxxxxDxxxxH. GxxxxDxAxxY is the active site, LxxxGxxxPxxGxG is the cofactor-binding site for NAD(P)(H), and LxxxxxxxxxDxxxxH is required for supporting the 3D structure
Comamonas testosteroni
Expression (protein specific)
Organism
Commentary
Expression
Comamonas testosteroni
the enzyme expression is significantly induced by estrone, estradiol, and methyltestosterone, and slightly by progesterone, but not by testosterone
up
Other publictions for EC 1.1.1.346
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)
742259
Chen
Cloning, expression and chara ...
Comamonas testosteroni, Comamonas testosteroni ATCC 11996
Chem. Biol. Interact.
234
229-235
2015
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1
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1
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4
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4
1
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1
3
3
1
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-
741575
Kaswurm
Evaluation of the food grade ...
Corynebacterium glutamicum, Corynebacterium glutamicum DSM 20301
AMB Express
3
7-17
2013
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1
1
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1
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2
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2
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6
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2
1
1
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1
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2
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1
1
2
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1
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2
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6
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2
1
1
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1
-
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-
2
2
-
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657308
Sanli
Structural alteration of cofac ...
Corynebacterium sp.
Protein Sci.
13
504-512
2004
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1
1
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1
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1
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1
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1
1
1
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1
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-
440310
Banta
Alteration of the specificity ...
Corynebacterium sp.
Protein Eng.
15
131-140
2002
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1
-
40
-
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1
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2
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1
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2
1
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1
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1
1
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40
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1
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1
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2
1
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440311
Banta
Optimizing an artificial metab ...
Corynebacterium sp.
Biochemistry
41
6226-6236
2002
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5
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1
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1
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5
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1
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1
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5
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1
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5
-
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-
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-
-
-
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440312
Habrych
Purification and identificatio ...
Escherichia coli, Escherichia coli BL21-(DE3)
Biotechnol. Prog.
18
257-261
2002
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1
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1
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2
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1
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1
1
5
1
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1
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1
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1
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1
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1
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1
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1
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1
1
5
1
-
-
-
1
-
-
-
1
-
1
1
-
-
-
440307
Khurana
Molecular modeling of substrat ...
Corynebacterium sp.
Proteins
39
68-75
2000
-
-
-
-
2
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1
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1
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2
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1
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440306
Yum
Identification of the yqhE and ...
Escherichia coli
Appl. Environ. Microbiol.
65
3341-3346
1999
-
-
-
-
-
-
-
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3
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8
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1
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2
1
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1
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3
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1
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2
1
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1
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-
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286278
Yum
Purification and characterizat ...
Brevibacterium ketosoreductum, Brevibacterium ketosoreductum ATCC 21914
Biosci. Biotechnol. Biochem.
62
154-156
1998
-
-
-
-
-
-
-
2
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2
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2
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1
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2
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5
1
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1
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1
1
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1
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1
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2
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2
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1
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2
-
5
1
-
-
-
-
1
-
1
1
-
-
-
-
-
-
440305
Khurana
Crystal structure of 2,5-diket ...
Corynebacterium sp.
Proc. Natl. Acad. Sci. USA
95
6768-6773
1998
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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
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1
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1
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-
721898
Maremonti
-
Characterisation of 2,5-diketo ...
Corynebacterium sp.
Biotechnol. Lett.
18
845-850
1996
-
-
-
-
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1
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2
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1
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1
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2
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1
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2
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1
1
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1
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1
-
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-
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-
-
440303
Sonoyama
-
Purification and properties of ...
Corynebacterium sp., Corynebacterium sp. SHS 0007
J. Ferment. Technol.
65
311-317
1987
-
-
-
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4
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2
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2
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1
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1
1
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1
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1
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5
1
1
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1
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2
-
1
2
-
-
-
-
-
-
440304
Miller
Purification and characterizat ...
Corynebacterium sp., Corynebacterium sp. ATCC 31090
J. Biol. Chem.
262
9016-9020
1987
-
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6
6
1
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2
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2
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1
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2
5
1
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1
2
1
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1
1
1
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1
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6
1
6
1
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2
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1
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2
2
5
1
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2
1
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1
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