BRENDA - Enzyme Database show
show all sequences of 1.1.99.21

A highly efficient sorbitol dehydrogenase from Gluconobacter oxydans G624 and improvement of its stability through immobilization

Kim, T.S.; Patel, S.K.; Selvaraj, C.; Jung, W.S.; Pan, C.H.; Kang, Y.C.; Lee, J.K.; Sci. Rep. 6, 33438 (2016)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
gene sldh, subcloning in Escherichia coli strain DH5alpha, recombinant expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)-CodonPlus RIL
Gluconobacter oxydans
Engineering
Amino acid exchange
Commentary
Organism
K294Q
site-directed mutagenesis, inactive mutant
Gluconobacter oxydans
additional information
stability of GoSLDH significantly improves up to 13.6fold after cross-linking of immobilized enzyme on silica nanoparticles and retains 62.8% residual activity after 10 cycles of reuse. Covalent immobilization of GoSLDH onto SiO2 nanoparticles: the amino groups of amino acids such as lysine present on the surface of GoSLDH react with the glutaraldehyde activates SiO2 nanoparticles to form covalent bonds during immobilization at pH 7. The IY and IE of GoSLDH immobilized on different silica nanoparticles are in the ranges of 40.4-71.2% and 53.5-76.7%, respectively
Gluconobacter oxydans
Inhibitors
Inhibitors
Commentary
Organism
Structure
Ba2+
-
Gluconobacter oxydans
Ca2+
-
Gluconobacter oxydans
Cu2+
-
Gluconobacter oxydans
K+
-
Gluconobacter oxydans
additional information
EDTA treatment does not affect the initial enzyme activity
Gluconobacter oxydans
Zn2+
-
Gluconobacter oxydans
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0882
-
NADP+
pH 10.0, 25C
Gluconobacter oxydans
38.9
-
D-sorbitol
pH 10.0, 25C
Gluconobacter oxydans
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
additional information
Mg2+, Co2+, and Mn2+ ions show no stimulatory influence on GoSLDH activity
Gluconobacter oxydans
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
53640
-
-
Gluconobacter oxydans
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
D-sorbitol + NADP+
Gluconobacter oxydans
-
L-sorbose + NADPH
-
-
r
D-sorbitol + NADP+
Gluconobacter oxydans G624
-
L-sorbose + NADPH
-
-
r
additional information
Gluconobacter oxydans
this SLDH is distinguished from other L-sorbose-producing enzymes by its high activity and substrate specificity. Isothermal titration calorimetry shows that the protein binds more strongly to D-sorbitol than other L-sorbose-producing enzymes, and substrate docking analysis confirms a higher turnover rate
?
-
-
-
additional information
Gluconobacter oxydans G624
this SLDH is distinguished from other L-sorbose-producing enzymes by its high activity and substrate specificity. Isothermal titration calorimetry shows that the protein binds more strongly to D-sorbitol than other L-sorbose-producing enzymes, and substrate docking analysis confirms a higher turnover rate
?
-
-
-
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Gluconobacter oxydans
-
-
-
Gluconobacter oxydans G624
-
-
-
Purification (Commentary)
Commentary
Organism
recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3)-CodonPlus RIL
Gluconobacter oxydans
Specific Activity [micromol/min/mg]
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
additional information
-
Gluconobacter oxydans G624 shows SLDH activity of 43.2 U/mL and a 20% conversion yield from D-sorbitol to L-sorbose at 18 h of fermentation
Gluconobacter oxydans
3570
-
purified recombinant enzyme, with D-sorbitol using NADP+ as a coenzyme, pH 10.0, 25C
Gluconobacter oxydans
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
D-sorbitol + NADP+
-
741426
Gluconobacter oxydans
L-sorbose + NADPH
-
-
-
r
D-sorbitol + NADP+
-
741426
Gluconobacter oxydans G624
L-sorbose + NADPH
-
-
-
r
additional information
this SLDH is distinguished from other L-sorbose-producing enzymes by its high activity and substrate specificity. Isothermal titration calorimetry shows that the protein binds more strongly to D-sorbitol than other L-sorbose-producing enzymes, and substrate docking analysis confirms a higher turnover rate
741426
Gluconobacter oxydans
?
-
-
-
-
additional information
GoSLDH is highly specific towards D-sorbitol, mannitol, and D-arabinitol (Table S2). No activity is detected with L-arabinitol, xylitol, ribitol, myo-inositol, and glycerol
741426
Gluconobacter oxydans
?
-
-
-
-
additional information
this SLDH is distinguished from other L-sorbose-producing enzymes by its high activity and substrate specificity. Isothermal titration calorimetry shows that the protein binds more strongly to D-sorbitol than other L-sorbose-producing enzymes, and substrate docking analysis confirms a higher turnover rate
741426
Gluconobacter oxydans G624
?
-
-
-
-
additional information
GoSLDH is highly specific towards D-sorbitol, mannitol, and D-arabinitol (Table S2). No activity is detected with L-arabinitol, xylitol, ribitol, myo-inositol, and glycerol
741426
Gluconobacter oxydans G624
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
dimer
-
Gluconobacter oxydans
Temperature Optimum [C]
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
70
-
oxidation reaction
Gluconobacter oxydans
Temperature Range [C]
Temperature Minimum [C]
Temperature Maximum [C]
Commentary
Organism
20
80
activity range, profile overview
Gluconobacter oxydans
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3820
-
D-sorbitol
pH 10.0, 25C
Gluconobacter oxydans
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
10
-
oxidation reaction
Gluconobacter oxydans
pH Range
pH Minimum
pH Maximum
Commentary
Organism
7.5
11
activity range, profile overview
Gluconobacter oxydans
Cofactor
Cofactor
Commentary
Organism
Structure
additional information
enzyme GoSLDH exhibits a preference for NADP+ over NAD+ as a coenzyme and is exclusively an NADP+-dependent enzyme showing only 2.5% relative activity with NAD+
Gluconobacter oxydans
NADP+
-
Gluconobacter oxydans
NADPH
-
Gluconobacter oxydans
Cloned(Commentary) (protein specific)
Commentary
Organism
gene sldh, subcloning in Escherichia coli strain DH5alpha, recombinant expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)-CodonPlus RIL
Gluconobacter oxydans
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
additional information
enzyme GoSLDH exhibits a preference for NADP+ over NAD+ as a coenzyme and is exclusively an NADP+-dependent enzyme showing only 2.5% relative activity with NAD+
Gluconobacter oxydans
NADP+
-
Gluconobacter oxydans
NADPH
-
Gluconobacter oxydans
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
K294Q
site-directed mutagenesis, inactive mutant
Gluconobacter oxydans
additional information
stability of GoSLDH significantly improves up to 13.6fold after cross-linking of immobilized enzyme on silica nanoparticles and retains 62.8% residual activity after 10 cycles of reuse. Covalent immobilization of GoSLDH onto SiO2 nanoparticles: the amino groups of amino acids such as lysine present on the surface of GoSLDH react with the glutaraldehyde activates SiO2 nanoparticles to form covalent bonds during immobilization at pH 7. The IY and IE of GoSLDH immobilized on different silica nanoparticles are in the ranges of 40.4-71.2% and 53.5-76.7%, respectively
Gluconobacter oxydans
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
Ba2+
-
Gluconobacter oxydans
Ca2+
-
Gluconobacter oxydans
Cu2+
-
Gluconobacter oxydans
K+
-
Gluconobacter oxydans
additional information
EDTA treatment does not affect the initial enzyme activity
Gluconobacter oxydans
Zn2+
-
Gluconobacter oxydans
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0882
-
NADP+
pH 10.0, 25C
Gluconobacter oxydans
38.9
-
D-sorbitol
pH 10.0, 25C
Gluconobacter oxydans
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
additional information
Mg2+, Co2+, and Mn2+ ions show no stimulatory influence on GoSLDH activity
Gluconobacter oxydans
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
53640
-
-
Gluconobacter oxydans
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
D-sorbitol + NADP+
Gluconobacter oxydans
-
L-sorbose + NADPH
-
-
r
D-sorbitol + NADP+
Gluconobacter oxydans G624
-
L-sorbose + NADPH
-
-
r
additional information
Gluconobacter oxydans
this SLDH is distinguished from other L-sorbose-producing enzymes by its high activity and substrate specificity. Isothermal titration calorimetry shows that the protein binds more strongly to D-sorbitol than other L-sorbose-producing enzymes, and substrate docking analysis confirms a higher turnover rate
?
-
-
-
additional information
Gluconobacter oxydans G624
this SLDH is distinguished from other L-sorbose-producing enzymes by its high activity and substrate specificity. Isothermal titration calorimetry shows that the protein binds more strongly to D-sorbitol than other L-sorbose-producing enzymes, and substrate docking analysis confirms a higher turnover rate
?
-
-
-
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3)-CodonPlus RIL
Gluconobacter oxydans
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
additional information
-
Gluconobacter oxydans G624 shows SLDH activity of 43.2 U/mL and a 20% conversion yield from D-sorbitol to L-sorbose at 18 h of fermentation
Gluconobacter oxydans
3570
-
purified recombinant enzyme, with D-sorbitol using NADP+ as a coenzyme, pH 10.0, 25C
Gluconobacter oxydans
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
D-sorbitol + NADP+
-
741426
Gluconobacter oxydans
L-sorbose + NADPH
-
-
-
r
D-sorbitol + NADP+
-
741426
Gluconobacter oxydans G624
L-sorbose + NADPH
-
-
-
r
additional information
this SLDH is distinguished from other L-sorbose-producing enzymes by its high activity and substrate specificity. Isothermal titration calorimetry shows that the protein binds more strongly to D-sorbitol than other L-sorbose-producing enzymes, and substrate docking analysis confirms a higher turnover rate
741426
Gluconobacter oxydans
?
-
-
-
-
additional information
GoSLDH is highly specific towards D-sorbitol, mannitol, and D-arabinitol (Table S2). No activity is detected with L-arabinitol, xylitol, ribitol, myo-inositol, and glycerol
741426
Gluconobacter oxydans
?
-
-
-
-
additional information
this SLDH is distinguished from other L-sorbose-producing enzymes by its high activity and substrate specificity. Isothermal titration calorimetry shows that the protein binds more strongly to D-sorbitol than other L-sorbose-producing enzymes, and substrate docking analysis confirms a higher turnover rate
741426
Gluconobacter oxydans G624
?
-
-
-
-
additional information
GoSLDH is highly specific towards D-sorbitol, mannitol, and D-arabinitol (Table S2). No activity is detected with L-arabinitol, xylitol, ribitol, myo-inositol, and glycerol
741426
Gluconobacter oxydans G624
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
dimer
-
Gluconobacter oxydans
Temperature Optimum [C] (protein specific)
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
70
-
oxidation reaction
Gluconobacter oxydans
Temperature Range [C] (protein specific)
Temperature Minimum [C]
Temperature Maximum [C]
Commentary
Organism
20
80
activity range, profile overview
Gluconobacter oxydans
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3820
-
D-sorbitol
pH 10.0, 25C
Gluconobacter oxydans
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
10
-
oxidation reaction
Gluconobacter oxydans
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
7.5
11
activity range, profile overview
Gluconobacter oxydans
General Information
General Information
Commentary
Organism
evolution
GoSLDH sequencing, structure analyses, and biochemical studies, suggest that it belongs to the NADP+-dependent polyol specific long-chain sorbitol dehydrogenase family. GoSLDH possesses the active site residues of Asp190, Val228, Lys294, and Asp299 and the conserved catalytic motif (KXXXXHXXH) for polyol-specific long-chain dehydrogenase
Gluconobacter oxydans
additional information
structure comparisons and molecular docking, overview
Gluconobacter oxydans
General Information (protein specific)
General Information
Commentary
Organism
evolution
GoSLDH sequencing, structure analyses, and biochemical studies, suggest that it belongs to the NADP+-dependent polyol specific long-chain sorbitol dehydrogenase family. GoSLDH possesses the active site residues of Asp190, Val228, Lys294, and Asp299 and the conserved catalytic motif (KXXXXHXXH) for polyol-specific long-chain dehydrogenase
Gluconobacter oxydans
additional information
structure comparisons and molecular docking, overview
Gluconobacter oxydans
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
98.1
-
D-sorbitol
pH 10.0, 25C
Gluconobacter oxydans
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
98.1
-
D-sorbitol
pH 10.0, 25C
Gluconobacter oxydans
Other publictions for EC 1.1.99.21
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)
739825
Fredslund
Structural characterization of ...
Bradyrhizobium japonicum, Bradyrhizobium japonicum USDA110
Acta Crystallogr. Sect. F
72
846-852
2016
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1
1
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4
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2
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4
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1
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740006
Selvaraj
-
Structural insights into the b ...
Gluconobacter oxydans
Biochem. Eng. J.
114
244-256
2016
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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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3
3
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741426
Kim
A highly efficient sorbitol de ...
Gluconobacter oxydans, Gluconobacter oxydans G624
Sci. Rep.
6
33438
2016
-
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1
-
2
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6
2
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1
1
4
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1
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6
1
1
1
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1
1
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2
2
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1
1
740961
Xu
Enhanced production of L-sorbo ...
Gluconobacter oxydans, Gluconobacter oxydans WSH-003
Microb. Cell Fact.
13
146
2014
-
1
1
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1
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1
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1
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1
1
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724595
Soemphol
Characterization of genes invo ...
Gluconobacter frateurii, Gluconobacter frateurii THD32
Biosci. Biotechnol. Biochem.
76
1497-1505
2012
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1
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1
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684722
Yang
A pyrroloquinoline quinine-dep ...
Gluconobacter oxydans
Arch. Biochem. Biophys.
477
206-210
2008
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1
4
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685690
Soemphol
Distinct physiological roles o ...
Gluconobacter frateurii
Biosci. Biotechnol. Biochem.
72
842-850
2008
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1
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695746
Yang
Membrane-bound pyrroloquinolin ...
Gluconobacter oxydans, Gluconobacter oxydans M5
Appl. Environ. Microbiol.
74
5250-5253
2008
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1
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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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1
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1
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672655
Toyama
Molecular properties of membra ...
Gluconobacter frateurii
Biosci. Biotechnol. Biochem.
69
1120-1129
2005
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654267
Matsushita
5-Keto-D-gluconate production ...
Gluconobacter oxydans
Appl. Environ. Microbiol.
69
1959-1966
2003
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1
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1
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1
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1
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8
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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
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-
1
-
-
-
-
-
-
-
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8
-
-
-
-
-
1
1
-
-
-
-
-
-
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389912
Shinagawa
-
Purification and characterizat ...
Gluconobacter oxydans
Agric. Biol. Chem.
46
135-141
1982
-
-
-
-
-
3
-
1
1
1
4
1
-
1
-
-
1
-
-
-
1
2
4
1
1
-
-
-
1
-
-
3
-
-
-
-
-
-
3
-
-
3
-
-
-
1
1
1
4
1
-
-
-
1
-
-
1
2
4
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
389913
Shinagawa
-
D-Sorbitol dehydrogenase from ...
Gluconobacter oxydans
Methods Enzymol.
89
141-145
1982
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-
-
-
-
-
-
1
1
-
4
1
-
1
-
-
1
-
-
-
1
-
4
1
1
-
-
-
1
-
-
3
-
-
-
-
-
-
3
-
-
-
-
-
-
1
1
-
4
1
-
-
-
1
-
-
1
-
4
1
1
-
-
-
1
-
-
-
-
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-
-
-
-
657542
Kinast
-
Vierstufige 1-Desoxynojirimyci ...
Gluconobacter oxydans
Angew. Chem.
93
799-800
1981
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1
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-
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1
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-
-
-
-
-
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1
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1
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1
-
-
-
-
-
-
-
1
-
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-
-
-
-
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-
-
-
-
-
-
-
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1
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1
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1
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