BRENDA - Enzyme Database show
show all sequences of 1.1.1.12

A single amino acid change (Y318F) in the L-arabitol dehydrogenase (LadA) from Aspergillus niger results in a significant increase in affinity for D-sorbitol

Rutten, L.; Ribot, C.; Trejo-Aguilar, B.; Woesten, H.A.; de Vries, R.P.; BMC Microbiol. 9, 166 (2009)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
derivatives of the expression vector pQE32 containing wild-type and mutated versions of ladA transformed to Escherichia coli M13 cells
Aspergillus niger
Crystallization (Commentary)
Crystallization
Organism
modelling of LadA on human SDH identifies residues M70 and Y318 in LadA, that may explain the absence of activity on D-sorbitol
Aspergillus niger
Engineering
Amino acid exchange
Commentary
Organism
M70F
almost complete loss of activity; results in a nearly complete enzyme inactivation
Aspergillus niger
Y318F
increased activity for L-arabinitol and xylitol, increased affinity for D-sorbitol; results in a significant increase in affinity for D-sorbitol, xylitol and L-arabitol
Aspergillus niger
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.056
-
L-arabinitol
wild-type, pH 9.6, 20°C
Aspergillus niger
0.056
-
L-arabitol
wild-type
Aspergillus niger
0.078
-
L-arabinitol
mutant Y318F, pH 9.6, 20°C
Aspergillus niger
0.078
-
L-arabitol
mutant Y318F
Aspergillus niger
0.218
-
xylitol
mutant Y318F; mutant Y318F, pH 9.6, 20°C
Aspergillus niger
0.25
-
xylitol
wild-type; wild-type, pH 9.6, 20°C
Aspergillus niger
0.868
-
D-sorbitol
mutant Y318F, pH 9.6, 20°C
Aspergillus niger
4.12
-
D-sorbitol
wild-type, pH 9.6, 20°C
Aspergillus niger
4.122
-
D-sorbitol
wild-type
Aspergillus niger
4.868
-
D-sorbitol
mutant Y318F
Aspergillus niger
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Aspergillus niger
A2QAC0
-
-
Purification (Commentary)
Commentary
Organism
recombinant proteins purified using Ni-agarose
Aspergillus niger
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
D-sorbitol + NAD+
-
696988
Aspergillus niger
D-fructose + NADH + H+
-
-
-
?
D-sorbitol + NAD+
wild-type shows no activity on D-sorbitol, whereas mutant Y318F does
696988
Aspergillus niger
? + NADH + H+
-
-
-
?
L-arabinitol + NAD+
-
696988
Aspergillus niger
L-xylulose + NADH + H+
-
-
-
?
L-arabitol + NAD+
-
696988
Aspergillus niger
L-xylulose + NADH + H+
-
-
-
?
additional information
Y318 of LadA contributes significantly to the substrate specificity difference between LAD and xylitol dehydrogenase/D-sorbitol dehydrogenase
696988
Aspergillus niger
?
-
-
-
-
xylitol + NAD+
-
696988
Aspergillus niger
D-xylulose + NADH + H+
-
-
-
?
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
13.48
-
D-sorbitol
wild-type, pH 9.6, 20°C
Aspergillus niger
13.88
-
D-sorbitol
mutant Y318F, pH 9.6, 20°C
Aspergillus niger
14.4
-
L-arabinitol
wild-type, pH 9.6, 20°C
Aspergillus niger
19.67
-
xylitol
wild-type, pH 9.6, 20°C
Aspergillus niger
30
-
L-arabinitol
mutant Y318F, pH 9.6, 20°C
Aspergillus niger
36.8
-
xylitol
mutant Y318F, pH 9.6, 20°C
Aspergillus niger
809
-
D-sorbitol
wild-type
Aspergillus niger
833
-
D-sorbitol
mutant Y318F
Aspergillus niger
863
-
L-arabitol
wild-type
Aspergillus niger
1180
-
xylitol
wild-type
Aspergillus niger
1800
-
L-arabitol
mutant Y318F
Aspergillus niger
2208
-
xylitol
mutant Y318F
Aspergillus niger
Cofactor
Cofactor
Commentary
Organism
Structure
NAD+
-
Aspergillus niger
Cloned(Commentary) (protein specific)
Commentary
Organism
derivatives of the expression vector pQE32 containing wild-type and mutated versions of ladA transformed to Escherichia coli M13 cells
Aspergillus niger
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NAD+
-
Aspergillus niger
Crystallization (Commentary) (protein specific)
Crystallization
Organism
modelling of LadA on human SDH identifies residues M70 and Y318 in LadA, that may explain the absence of activity on D-sorbitol
Aspergillus niger
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
M70F
almost complete loss of activity; results in a nearly complete enzyme inactivation
Aspergillus niger
Y318F
increased activity for L-arabinitol and xylitol, increased affinity for D-sorbitol; results in a significant increase in affinity for D-sorbitol, xylitol and L-arabitol
Aspergillus niger
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.056
-
L-arabinitol
wild-type, pH 9.6, 20°C
Aspergillus niger
0.056
-
L-arabitol
wild-type
Aspergillus niger
0.078
-
L-arabinitol
mutant Y318F, pH 9.6, 20°C
Aspergillus niger
0.078
-
L-arabitol
mutant Y318F
Aspergillus niger
0.218
-
xylitol
mutant Y318F; mutant Y318F, pH 9.6, 20°C
Aspergillus niger
0.25
-
xylitol
wild-type; wild-type, pH 9.6, 20°C
Aspergillus niger
0.868
-
D-sorbitol
mutant Y318F, pH 9.6, 20°C
Aspergillus niger
4.12
-
D-sorbitol
wild-type, pH 9.6, 20°C
Aspergillus niger
4.122
-
D-sorbitol
wild-type
Aspergillus niger
4.868
-
D-sorbitol
mutant Y318F
Aspergillus niger
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant proteins purified using Ni-agarose
Aspergillus niger
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
D-sorbitol + NAD+
-
696988
Aspergillus niger
D-fructose + NADH + H+
-
-
-
?
D-sorbitol + NAD+
wild-type shows no activity on D-sorbitol, whereas mutant Y318F does
696988
Aspergillus niger
? + NADH + H+
-
-
-
?
L-arabinitol + NAD+
-
696988
Aspergillus niger
L-xylulose + NADH + H+
-
-
-
?
L-arabitol + NAD+
-
696988
Aspergillus niger
L-xylulose + NADH + H+
-
-
-
?
additional information
Y318 of LadA contributes significantly to the substrate specificity difference between LAD and xylitol dehydrogenase/D-sorbitol dehydrogenase
696988
Aspergillus niger
?
-
-
-
-
xylitol + NAD+
-
696988
Aspergillus niger
D-xylulose + NADH + H+
-
-
-
?
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
13.48
-
D-sorbitol
wild-type, pH 9.6, 20°C
Aspergillus niger
13.88
-
D-sorbitol
mutant Y318F, pH 9.6, 20°C
Aspergillus niger
14.4
-
L-arabinitol
wild-type, pH 9.6, 20°C
Aspergillus niger
19.67
-
xylitol
wild-type, pH 9.6, 20°C
Aspergillus niger
30
-
L-arabinitol
mutant Y318F, pH 9.6, 20°C
Aspergillus niger
36.8
-
xylitol
mutant Y318F, pH 9.6, 20°C
Aspergillus niger
809
-
D-sorbitol
wild-type
Aspergillus niger
833
-
D-sorbitol
mutant Y318F
Aspergillus niger
863
-
L-arabitol
wild-type
Aspergillus niger
1180
-
xylitol
wild-type
Aspergillus niger
1800
-
L-arabitol
mutant Y318F
Aspergillus niger
2208
-
xylitol
mutant Y318F
Aspergillus niger
Other publictions for EC 1.1.1.12
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)
710970
Kim
Cloning, characterization, and ...
Aspergillus niger, Penicillium chrysogenum, Penicillium chrysogenum Wisconsin 54-1255, Trichoderma reesei
Appl. Microbiol. Biotechnol.
87
1407-1414
2010
-
-
-
-
3
-
-
6
-
-
-
-
-
4
-
-
-
-
-
-
3
-
4
-
3
-
-
6
-
-
-
6
-
-
-
-
-
-
6
-
3
-
-
-
-
6
-
-
-
-
-
-
-
-
-
-
3
-
4
-
3
-
-
6
-
-
-
-
-
-
-
-
6
6
711121
Fernandes
Cloning, heterologous expressi ...
Rasamsonia emersonii
Biochem. Genet.
48
480-495
2010
-
-
1
-
-
-
-
4
-
1
2
-
-
1
-
-
-
-
-
-
-
-
3
1
-
1
-
4
1
1
-
2
-
-
-
-
-
1
2
-
-
-
-
-
-
4
-
1
2
-
-
-
-
-
-
-
-
-
3
1
-
1
-
4
1
1
-
-
-
-
-
-
4
4
712767
Bae
Structure and engineering of L ...
Neurospora crassa
J. Mol. Biol.
402
230-240
2010
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9
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1
-
-
-
-
-
5
-
-
-
-
6
-
-
-
1
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-
-
-
-
-
1
1
4
-
-
-
-
9
-
1
-
-
-
-
-
1
-
-
-
-
5
-
-
-
-
6
-
-
-
-
-
-
-
-
6
6
712797
Tiwari
Molecular modeling studies of ...
Trichoderma reesei
J. Mol. Graph. Model.
28
707-713
2010
-
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1
-
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1
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-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
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-
-
1
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-
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1
1
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1
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-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
696988
Rutten
A single amino acid change (Y3 ...
Aspergillus niger
BMC Microbiol.
9
166
2009
-
-
1
1
2
-
-
10
-
-
-
-
-
3
-
-
1
-
-
-
-
-
6
-
-
-
-
12
-
-
-
1
-
-
-
-
-
1
1
1
2
-
-
-
-
10
-
-
-
-
-
-
-
1
-
-
-
-
6
-
-
-
-
12
-
-
-
-
-
-
-
-
-
-
711877
Akel
Molecular regulation of arabin ...
Trichoderma reesei
Eukaryot. Cell
8
1837-1844
2009
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
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-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
1
-
-
-
695737
Fonseca
Use of in vivo 13C nuclear mag ...
Candida arabinofermentans, Candida arabinofermentans PYCC 5603T, Meyerozyma guilliermondii, Meyerozyma guilliermondii PYCC 3012
Appl. Environ. Microbiol.
74
1845-1855
2008
-
-
-
-
-
-
-
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-
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-
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-
5
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4
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2
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-
2
-
-
-
-
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-
-
-
-
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
684627
Sullivan
Cloning, characterization, and ...
Neurospora crassa
Appl. Microbiol. Biotechnol.
77
845-852
2007
-
1
1
-
-
-
-
4
-
1
3
-
-
3
-
-
1
-
-
-
-
2
4
1
1
-
2
4
1
1
-
4
-
-
-
-
1
1
4
-
-
-
-
-
-
4
-
1
3
-
-
-
-
1
-
-
-
2
4
1
1
-
2
4
1
1
-
-
-
-
-
-
-
-
686661
Fonseca
L-Arabinose transport and cata ...
Candida arabinofermentans, Candida arabinofermentans PYCC, Meyerozyma guilliermondii, Meyerozyma guilliermondii PYCC 3012
FEBS J.
274
3589-3600
2007
-
-
-
-
-
-
-
2
-
-
-
-
-
5
-
-
-
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-
2
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4
-
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-
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-
-
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2
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-
2
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
668063
de Groot
Metabolic control analysis of ...
Aspergillus niger
Biotechnol. Prog.
21
1610-1616
2005
-
-
-
-
-
-
-
4
-
-
-
1
-
2
-
-
1
-
-
-
-
2
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
2
-
-
-
-
-
-
4
-
-
-
1
-
-
-
1
-
-
-
2
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
669541
Suzuki
Cloning and expression of NAD+ ...
Aspergillus oryzae
J. Biosci. Bioeng.
100
472-474
2005
-
-
1
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4
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3
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1
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1
1
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3
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655480
Pail
The metabolic role and evoluti ...
Trichoderma reesei
Eur. J. Biochem.
271
1864-1872
2004
-
-
-
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1
-
-
14
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6
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1
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-
-
-
14
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-
1
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-
14
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-
-
1
-
-
-
-
14
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
655390
Seiboth
D-xylose metabolism in Hypocre ...
Trichoderma reesei
Eukaryot. Cell
2
867-875
2003
-
-
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1
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2
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2
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285788
De Vries
Characterization of an Aspergi ...
Aspergillus nidulans
FEMS Microbiol. Lett.
123
83-90
1994
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-
-
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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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285787
Witteveen
-
L-Arabinose and D-xylose catab ...
Aspergillus niger
J. Gen. Microbiol.
135
2163-2171
1989
-
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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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285789
Chiang
L-Arabinose metabolism by cell ...
Penicillium chrysogenum
Biochim. Biophys. Acta
46
271-278
1961
-
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1
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1
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1
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246416
Chiang
A new pathway of pentose metab ...
Penicillium chrysogenum
Biochem. Biophys. Res. Commun.
3
554-559
1960
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