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show all sequences of 1.1.1.292

Crystal structure of NADP(H)-dependent 1,5-anhydro-D-fructose reductase from Sinorhizobium morelense at 2.2 A resolution: construction of a NADH-accepting mutant and its application in rare sugar synthesis

Dambe, T.R.; Kuehn, A.M.; Brossette, T.; Giffhorn, F.; Scheidig, A.J.; Biochemistry 45, 10030-10042 (2006)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expression in Escherichia coli
Ensifer adhaerens
Crystallization (Commentary)
Crystallization
Organism
hanging drop vapor diffusion method, enzyme crystallized in complex with the cofactor NADP(H) and its structure is determined to 2.2 A resolution using selenomethionine single-wavelength anomalous dispersion
Ensifer adhaerens
Engineering
Amino acid exchange
Commentary
Organism
A13G
kcat/Km for reaction with 1,5-anhydro-D-fructose and NADPH is 1.8fold higher than wild type value. Mutant enzyme shows activity with NADH as cofactor
Ensifer adhaerens
G206I
kcat/Km for reaction with 1,5-anhydro-D-fructose and NADPH is 1.4fold lower than wild type value
Ensifer adhaerens
H180A
kcat/Km for reaction with 1,5-anhydro-D-fructose and NADPH is 61.9fold lower than wild type value
Ensifer adhaerens
K94G
kcat/Km for reaction with 1,5-anhydro-D-fructose and NADPH is 137fold lower than wild type value
Ensifer adhaerens
S10G
kcat/Km for reaction with 1,5-anhydro-D-fructose and NADPH is 1.3fold higher than wild type value
Ensifer adhaerens
S10G/A13G
kcat/Km for reaction with 1,5-anhydro-D-fructose and NADPH is 2fold higher than wild type value. Mutant enzyme shows activity with NADH as cofactor
Ensifer adhaerens
S176A
kcat/Km for reaction with 1,5-anhydro-D-fructose and NADPH is 1001fold lower than wild type value
Ensifer adhaerens
S33D
no activity
Ensifer adhaerens
S33D/A13G
kcat/Km for reaction with 1,5-anhydro-D-fructose and NADPH is 84fold lower than wild type value. Mutant enzyme shows activity with NADH as cofactor
Ensifer adhaerens
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.02
-
NADPH
pH 6.5, mutant enzyme A13G
Ensifer adhaerens
0.06
-
NADPH
pH 6.5, mutant enzyme G206I; pH 6.5 recombinant wild-type enzyme
Ensifer adhaerens
0.1
-
NADPH
pH 6.5, native wild-type enzyme
Ensifer adhaerens
0.2
-
NADPH
pH 6.5, mutant enzyme K94G
Ensifer adhaerens
0.27
-
NADPH
pH 6.5, mutant enzyme S10G
Ensifer adhaerens
0.38
-
NADPH
pH 6.5, mutant enzyme S10G/A13G
Ensifer adhaerens
1
-
NADPH
pH 6.5, mutant enzyme S33D/A13G
Ensifer adhaerens
1.1
-
NADH
pH 6.5, mutant enzyme A13G; pH 6.5, mutant enzyme S33D/A13G
Ensifer adhaerens
1.2
-
NADH
pH 6.5, mutant enzyme S10G/A13G
Ensifer adhaerens
3.2
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme S33D/A13G, cofactor: NADH
Ensifer adhaerens
3.5
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme S10G, cofactor: NADPH
Ensifer adhaerens
6.4
-
1,5-Anhydro-D-fructose
pH 6.5 recombinant wild-type enzyme, cofactor: NADPH
Ensifer adhaerens
7.1
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme S10G/A13G, cofactor: NADPH
Ensifer adhaerens
8.3
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme G206I, cofactor: NADPH; pH 6.5, native wild-type enzyme, cofactor: NADPH
Ensifer adhaerens
8.5
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme A13G, cofactor: NADPH
Ensifer adhaerens
8.9
-
1,5-Anhydro-D-fructose
pH 8.0, mutant enzyme H180A, cofactor: NADPH
Ensifer adhaerens
11.1
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme A13G, cofactor: NADH
Ensifer adhaerens
20.2
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme S33D/A13G, cofactor: NADPH
Ensifer adhaerens
22.5
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme K94G, cofactor: NADPH
Ensifer adhaerens
39
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme S10G/A13G, cofactor: NADH
Ensifer adhaerens
49
-
1,5-Anhydro-D-fructose
pH 7.5, mutant enzyme D176A, cofactor: NADPH
Ensifer adhaerens
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Ensifer adhaerens
Q2I8V6
i.e. Ensifer adhaerens
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1,5-anhydro-D-fructose + NADPH + H+
-
667698
Ensifer adhaerens
1,5-anhydro-D-mannitol + NADP+
-
-
-
?
1,5-anhydro-D-fructose + NADPH + H+
-
667698
Ensifer adhaerens S-30.7.5
1,5-anhydro-D-mannitol + NADP+
-
-
-
?
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.3
-
1,5-Anhydro-D-fructose
pH 7.5, mutant enzyme D176A, cofactor: NADPH
Ensifer adhaerens
3.7
-
1,5-Anhydro-D-fructose
pH 8.0, mutant enzyme H180A, cofactor: NADPH
Ensifer adhaerens
4.2
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme K94G, cofactor: NADPH
Ensifer adhaerens
5.5
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme S10G/A13G, cofactor: NADH
Ensifer adhaerens
6.3
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme S33D/A13G, cofactor: NADPH
Ensifer adhaerens
12.4
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme A13G, cofactor: NADH
Ensifer adhaerens
13.5
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme S33D/A13G, cofactor: NADH
Ensifer adhaerens
119
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme S10G, cofactor: NADPH
Ensifer adhaerens
145
-
1,5-Anhydro-D-fructose
pH 6.5 recombinant wild-type enzyme, cofactor: NADPH
Ensifer adhaerens
156
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme G206I, cofactor: NADPH
Ensifer adhaerens
216
-
1,5-Anhydro-D-fructose
pH 6.5, native wild-type enzyme, cofactor: NADPH
Ensifer adhaerens
369
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme S10G/A13G, cofactor: NADPH
Ensifer adhaerens
405
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme A13G, cofactor: NADPH
Ensifer adhaerens
Cofactor
Cofactor
Commentary
Organism
Structure
NADH
wild-type enzyme and mutant enzymes S10G, S33D, K94G, D176A, H180A and G206I shows no activity with NADH. Mutant enzymes A13G, S10G/A13G and S33D/A13G are active with NADH
Ensifer adhaerens
NADPH
the N-terminal domain displays a Rossman fold and contains the cofactor binding site. The intact crystals contain the oxidized cofactor NADP+
Ensifer adhaerens
Cloned(Commentary) (protein specific)
Commentary
Organism
expression in Escherichia coli
Ensifer adhaerens
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NADH
wild-type enzyme and mutant enzymes S10G, S33D, K94G, D176A, H180A and G206I shows no activity with NADH. Mutant enzymes A13G, S10G/A13G and S33D/A13G are active with NADH
Ensifer adhaerens
NADPH
the N-terminal domain displays a Rossman fold and contains the cofactor binding site. The intact crystals contain the oxidized cofactor NADP+
Ensifer adhaerens
Crystallization (Commentary) (protein specific)
Crystallization
Organism
hanging drop vapor diffusion method, enzyme crystallized in complex with the cofactor NADP(H) and its structure is determined to 2.2 A resolution using selenomethionine single-wavelength anomalous dispersion
Ensifer adhaerens
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
A13G
kcat/Km for reaction with 1,5-anhydro-D-fructose and NADPH is 1.8fold higher than wild type value. Mutant enzyme shows activity with NADH as cofactor
Ensifer adhaerens
G206I
kcat/Km for reaction with 1,5-anhydro-D-fructose and NADPH is 1.4fold lower than wild type value
Ensifer adhaerens
H180A
kcat/Km for reaction with 1,5-anhydro-D-fructose and NADPH is 61.9fold lower than wild type value
Ensifer adhaerens
K94G
kcat/Km for reaction with 1,5-anhydro-D-fructose and NADPH is 137fold lower than wild type value
Ensifer adhaerens
S10G
kcat/Km for reaction with 1,5-anhydro-D-fructose and NADPH is 1.3fold higher than wild type value
Ensifer adhaerens
S10G/A13G
kcat/Km for reaction with 1,5-anhydro-D-fructose and NADPH is 2fold higher than wild type value. Mutant enzyme shows activity with NADH as cofactor
Ensifer adhaerens
S176A
kcat/Km for reaction with 1,5-anhydro-D-fructose and NADPH is 1001fold lower than wild type value
Ensifer adhaerens
S33D
no activity
Ensifer adhaerens
S33D/A13G
kcat/Km for reaction with 1,5-anhydro-D-fructose and NADPH is 84fold lower than wild type value. Mutant enzyme shows activity with NADH as cofactor
Ensifer adhaerens
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.02
-
NADPH
pH 6.5, mutant enzyme A13G
Ensifer adhaerens
0.06
-
NADPH
pH 6.5, mutant enzyme G206I; pH 6.5 recombinant wild-type enzyme
Ensifer adhaerens
0.1
-
NADPH
pH 6.5, native wild-type enzyme
Ensifer adhaerens
0.2
-
NADPH
pH 6.5, mutant enzyme K94G
Ensifer adhaerens
0.27
-
NADPH
pH 6.5, mutant enzyme S10G
Ensifer adhaerens
0.38
-
NADPH
pH 6.5, mutant enzyme S10G/A13G
Ensifer adhaerens
1
-
NADPH
pH 6.5, mutant enzyme S33D/A13G
Ensifer adhaerens
1.1
-
NADH
pH 6.5, mutant enzyme A13G; pH 6.5, mutant enzyme S33D/A13G
Ensifer adhaerens
1.2
-
NADH
pH 6.5, mutant enzyme S10G/A13G
Ensifer adhaerens
3.2
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme S33D/A13G, cofactor: NADH
Ensifer adhaerens
3.5
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme S10G, cofactor: NADPH
Ensifer adhaerens
6.4
-
1,5-Anhydro-D-fructose
pH 6.5 recombinant wild-type enzyme, cofactor: NADPH
Ensifer adhaerens
7.1
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme S10G/A13G, cofactor: NADPH
Ensifer adhaerens
8.3
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme G206I, cofactor: NADPH; pH 6.5, native wild-type enzyme, cofactor: NADPH
Ensifer adhaerens
8.5
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme A13G, cofactor: NADPH
Ensifer adhaerens
8.9
-
1,5-Anhydro-D-fructose
pH 8.0, mutant enzyme H180A, cofactor: NADPH
Ensifer adhaerens
11.1
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme A13G, cofactor: NADH
Ensifer adhaerens
20.2
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme S33D/A13G, cofactor: NADPH
Ensifer adhaerens
22.5
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme K94G, cofactor: NADPH
Ensifer adhaerens
39
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme S10G/A13G, cofactor: NADH
Ensifer adhaerens
49
-
1,5-Anhydro-D-fructose
pH 7.5, mutant enzyme D176A, cofactor: NADPH
Ensifer adhaerens
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1,5-anhydro-D-fructose + NADPH + H+
-
667698
Ensifer adhaerens
1,5-anhydro-D-mannitol + NADP+
-
-
-
?
1,5-anhydro-D-fructose + NADPH + H+
-
667698
Ensifer adhaerens S-30.7.5
1,5-anhydro-D-mannitol + NADP+
-
-
-
?
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.3
-
1,5-Anhydro-D-fructose
pH 7.5, mutant enzyme D176A, cofactor: NADPH
Ensifer adhaerens
3.7
-
1,5-Anhydro-D-fructose
pH 8.0, mutant enzyme H180A, cofactor: NADPH
Ensifer adhaerens
4.2
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme K94G, cofactor: NADPH
Ensifer adhaerens
5.5
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme S10G/A13G, cofactor: NADH
Ensifer adhaerens
6.3
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme S33D/A13G, cofactor: NADPH
Ensifer adhaerens
12.4
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme A13G, cofactor: NADH
Ensifer adhaerens
13.5
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme S33D/A13G, cofactor: NADH
Ensifer adhaerens
119
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme S10G, cofactor: NADPH
Ensifer adhaerens
145
-
1,5-Anhydro-D-fructose
pH 6.5 recombinant wild-type enzyme, cofactor: NADPH
Ensifer adhaerens
156
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme G206I, cofactor: NADPH
Ensifer adhaerens
216
-
1,5-Anhydro-D-fructose
pH 6.5, native wild-type enzyme, cofactor: NADPH
Ensifer adhaerens
369
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme S10G/A13G, cofactor: NADPH
Ensifer adhaerens
405
-
1,5-Anhydro-D-fructose
pH 6.5, mutant enzyme A13G, cofactor: NADPH
Ensifer adhaerens
Other publictions for EC 1.1.1.292
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)
739808
Schu
The structure of substrate-fre ...
Sinorhizobium meliloti
Acta Crystallogr. Sect. F
69
844-849
2013
-
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1
1
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-
-
-
-
-
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1
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-
-
-
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-
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-
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-
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1
-
1
-
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-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
685692
Sakuma
Mouse AKR1E1 is an ortholog of ...
Mus musculus, Mus musculus C57/BL6J
Biosci. Biotechnol. Biochem.
72
872-876
2008
-
-
1
-
-
-
-
1
-
-
-
2
-
2
-
-
1
-
-
-
-
-
10
-
1
-
-
1
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
1
-
-
-
2
-
-
-
1
-
-
-
-
10
-
1
-
-
1
1
-
-
-
-
-
-
-
-
-
667264
Kuehn
Catabolism of 1,5-anhydro-D-fr ...
Ensifer adhaerens, Ensifer adhaerens S-30.7.5
Appl. Environ. Microbiol.
72
1248-1257
2006
-
-
1
-
-
-
-
3
-
-
3
-
-
2
-
-
1
-
-
-
2
1
11
1
-
-
-
3
1
1
-
2
-
1
-
-
-
1
2
-
-
-
-
-
-
3
-
-
3
-
-
-
-
1
-
-
2
1
11
1
-
-
-
3
1
1
-
1
-
-
-
-
-
-
667698
Dambe
Crystal structure of NADP(H)-d ...
Ensifer adhaerens
Biochemistry
45
10030-10042
2006
-
-
1
1
9
-
-
21
-
-
-
-
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
13
-
-
-
2
-
-
-
-
-
1
2
1
9
-
-
-
-
21
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
13
-
-
-
-
-
-
-
-
-
-