BRENDA - Enzyme Database show
show all sequences of 1.14.14.92

Purification and properties of benzoate-4-hydroxylase from a soil pseudomonad

Reddy, C.C.; Vaidyanathan, C.S.; Arch. Biochem. Biophys. 177, 488-498 (1976)

Data extracted from this reference:

Activating Compound
Activating Compound
Commentary
Organism
Structure
2-mercaptoethanol
hydroxylation is not supported when NADH, NADPH, dithiothreitol, 2-mercaptoethanol and ascorbic acid are added alone to the reaction mixture, stimulation when added together with tetrahydropteridine
Pseudomonas sp.
ascorbic acid
hydroxylation is not supported when NADH, NADPH, dithiothreitol, 2-mercaptoethanol and ascorbic acid are added alone to the reaction mixture, stimulation when added together with tetrahydropteridine
Pseudomonas sp.
dithiothreitol
hydroxylation is not supported when NADH, NADPH, dithiothreitol, 2-mercaptoethanol and ascorbic acid are added alone to the reaction mixture, stimulation when added together with tetrahydropteridine
Pseudomonas sp.
General Stability
General Stability
Organism
freezing and thawing has no effect
Pseudomonas sp.
quite stable in presence of benzoate
Pseudomonas sp.
Inhibitors
Inhibitors
Commentary
Organism
Structure
4-Chlorobenzoate
-
Pseudomonas sp.
4-Methylbenzoate
-
Pseudomonas sp.
4-nitrobenzoate
38% inhibition
Pseudomonas sp.
benzaldehyde
-
Pseudomonas sp.
benzoate methyl ester
slight inhibition
Pseudomonas sp.
Cu2+
0.005 mM, 50% inhibition
Pseudomonas sp.
Hg2+
0.005 mM, 70% inhibition
Pseudomonas sp.
m-Hydroxybenzoate
12% inhibition
Pseudomonas sp.
salicylate
20% inhibition
Pseudomonas sp.
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.045
-
6,7-dimethyltetrahydropterin
-
Pseudomonas sp.
0.045
-
NADH
-
Pseudomonas sp.
0.3
-
Benzoate
-
Pseudomonas sp.
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Fe2+
preincubation for 10 min in the presence of benzoate results in maximal activity
Pseudomonas sp.
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
120000
-
gel filtration
Pseudomonas sp.
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Pseudomonas sp.
-
-
-
Purification (Commentary)
Commentary
Organism
protamine sulfate, heat treatment, calcium phosphate gel, ammonium sulfate, DEAE-Sephadex, Sephadex G-150
Pseudomonas sp.
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
33.92
-
-
Pseudomonas sp.
Storage Stability
Storage Stability
Organism
0-4°C, enzyme concentration 0.050 mg/ml, pH 6.0-7.5, 15 days, no loss of activity
Pseudomonas sp.
0-4°C, pH 7.2-8.0, Tris-HCl buffer, 24 h, 50% loss of activity
Pseudomonas sp.
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4-chlorobenzoate + [reduced NADPH-hemoprotein reductase] + O2
-
440255
Pseudomonas sp.
?
-
-
-
?
4-methylbenzoate + [reduced NADPH-hemoprotein reductase] + O2
-
440255
Pseudomonas sp.
?
-
-
-
?
benzoate + [reduced NADPH-hemoprotein reductase] + O2
-
440255
Pseudomonas sp.
4-hydroxybenzoate + [oxidized NADPH-hemoprotein reductase] + H2O
-
440255
Pseudomonas sp.
?
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
34
-
-
Pseudomonas sp.
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.2
-
-
Pseudomonas sp.
pH Range
pH Minimum
pH Maximum
Commentary
Organism
5.5
8.2
approx. 30% of maximal activity at pH 5.5, approx. 35% of maximal activity at pH 8.2
Pseudomonas sp.
Cofactor
Cofactor
Commentary
Organism
Structure
tetrahydropteridine
essential for activity, hydroxylation is not supported when NADH, NADPH, dithiothreitol, 2-mercaptoethanol and ascorbic acid are added alone to the reaction mixture, stimulation when added together with tetrahydropteridine
Pseudomonas sp.
Ki Value [mM]
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.75
-
4-nitrobenzoate
-
Pseudomonas sp.
Activating Compound (protein specific)
Activating Compound
Commentary
Organism
Structure
2-mercaptoethanol
hydroxylation is not supported when NADH, NADPH, dithiothreitol, 2-mercaptoethanol and ascorbic acid are added alone to the reaction mixture, stimulation when added together with tetrahydropteridine
Pseudomonas sp.
ascorbic acid
hydroxylation is not supported when NADH, NADPH, dithiothreitol, 2-mercaptoethanol and ascorbic acid are added alone to the reaction mixture, stimulation when added together with tetrahydropteridine
Pseudomonas sp.
dithiothreitol
hydroxylation is not supported when NADH, NADPH, dithiothreitol, 2-mercaptoethanol and ascorbic acid are added alone to the reaction mixture, stimulation when added together with tetrahydropteridine
Pseudomonas sp.
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
tetrahydropteridine
essential for activity, hydroxylation is not supported when NADH, NADPH, dithiothreitol, 2-mercaptoethanol and ascorbic acid are added alone to the reaction mixture, stimulation when added together with tetrahydropteridine
Pseudomonas sp.
General Stability (protein specific)
General Stability
Organism
freezing and thawing has no effect
Pseudomonas sp.
quite stable in presence of benzoate
Pseudomonas sp.
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
4-Chlorobenzoate
-
Pseudomonas sp.
4-Methylbenzoate
-
Pseudomonas sp.
4-nitrobenzoate
38% inhibition
Pseudomonas sp.
benzaldehyde
-
Pseudomonas sp.
benzoate methyl ester
slight inhibition
Pseudomonas sp.
Cu2+
0.005 mM, 50% inhibition
Pseudomonas sp.
Hg2+
0.005 mM, 70% inhibition
Pseudomonas sp.
m-Hydroxybenzoate
12% inhibition
Pseudomonas sp.
salicylate
20% inhibition
Pseudomonas sp.
Ki Value [mM] (protein specific)
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.75
-
4-nitrobenzoate
-
Pseudomonas sp.
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.045
-
6,7-dimethyltetrahydropterin
-
Pseudomonas sp.
0.045
-
NADH
-
Pseudomonas sp.
0.3
-
Benzoate
-
Pseudomonas sp.
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Fe2+
preincubation for 10 min in the presence of benzoate results in maximal activity
Pseudomonas sp.
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
120000
-
gel filtration
Pseudomonas sp.
Purification (Commentary) (protein specific)
Commentary
Organism
protamine sulfate, heat treatment, calcium phosphate gel, ammonium sulfate, DEAE-Sephadex, Sephadex G-150
Pseudomonas sp.
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
33.92
-
-
Pseudomonas sp.
Storage Stability (protein specific)
Storage Stability
Organism
0-4°C, enzyme concentration 0.050 mg/ml, pH 6.0-7.5, 15 days, no loss of activity
Pseudomonas sp.
0-4°C, pH 7.2-8.0, Tris-HCl buffer, 24 h, 50% loss of activity
Pseudomonas sp.
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4-chlorobenzoate + [reduced NADPH-hemoprotein reductase] + O2
-
440255
Pseudomonas sp.
?
-
-
-
?
4-methylbenzoate + [reduced NADPH-hemoprotein reductase] + O2
-
440255
Pseudomonas sp.
?
-
-
-
?
benzoate + [reduced NADPH-hemoprotein reductase] + O2
-
440255
Pseudomonas sp.
4-hydroxybenzoate + [oxidized NADPH-hemoprotein reductase] + H2O
-
440255
Pseudomonas sp.
?
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
34
-
-
Pseudomonas sp.
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.2
-
-
Pseudomonas sp.
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
5.5
8.2
approx. 30% of maximal activity at pH 5.5, approx. 35% of maximal activity at pH 8.2
Pseudomonas sp.
Other publictions for EC 1.14.14.92
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)
741716
Antunes
A benzoate-activated promoter ...
Aspergillus niger
Appl. Microbiol. Biotechnol.
100
5479-5489
2016
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1
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1
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1
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742764
Korosec
Antifungal activity of cinnam ...
Curvularia lunata
J. Appl. Microbiol.
116
955-966
2014
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2
1
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1
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5
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1
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2
1
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1
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1
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2
2
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725603
Berne
Virtual screening yields inhib ...
Curvularia lunata
J. Chem. Inf. Model.
52
3053-3063
2012
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1
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686803
Kim
Molecular cloning and function ...
Chromohalobacter sp. HS2
FEMS Microbiol. Lett.
280
235-241
2008
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1
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1
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1
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3
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1
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671533
Shiningavamwe
Heterologous expression of the ...
Cystobasidium minutum, Cystobasidium minutum CBS 2177
Appl. Microbiol. Biotechnol.
72
323-329
2006
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1
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2
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6
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1
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1
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1
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2
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671704
Matsuzaki
Molecular characterization of ...
Phanerochaete chrysosporium
Biochem. Biophys. Res. Commun.
334
1184-1190
2005
1
-
1
-
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1
1
1
1
-
1
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5
1
1
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1
-
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1
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1
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1
1
1
1
-
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5
1
1
-
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-
1
-
-
-
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-
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440261
Faber
Purification and characterizat ...
Aspergillus niger
Arch. Biochem. Biophys.
394
245-254
2001
-
-
-
-
-
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-
9
-
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1
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1
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12
-
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13
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9
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1
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12
-
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13
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440259
Fukuda
Cytochrome P450rm from Rhodoto ...
Cystobasidium minutum
J. Biochem.
119
314-318
1996
-
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3
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1
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4
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2
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3
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1
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1
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3
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1
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2
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3
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440260
Van Den Brink
Optimization of the benzoate-i ...
Aspergillus niger, Aspergillus niger N204
Appl. Microbiol. Biotechnol.
46
360-364
1996
-
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1
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2
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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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-
-
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440254
McNamee
Properties of a membrane-assoc ...
Rhodotorula graminis
Biochem. Biophys. Res. Commun.
129
485-492
1985
-
-
1
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-
15
2
1
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3
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1
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1
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1
2
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15
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2
1
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1
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1
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4804
Durham
Initial reactions involved in ...
Rhodotorula graminis
J. Bacteriol.
160
778-780
1984
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1
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1
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440258
Shailubhai
-
Degradation of chlorinated der ...
Aspergillus niger
FEMS Microbiol. Lett.
18
279-282
1983
-
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1
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1
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1
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1
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440255
Reddy
Purification and properties of ...
Pseudomonas sp.
Arch. Biochem. Biophys.
177
488-498
1976
3
-
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-
2
9
3
-
1
1
-
-
1
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1
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1
2
3
-
1
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1
1
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1
1
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3
-
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1
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2
-
9
1
3
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1
1
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1
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1
2
3
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1
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1
1
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440256
Reddy
Purification, properties and i ...
Aspergillus niger, Aspergillus niger UBC 814
Biochim. Biophys. Acta
384
46-57
1975
1
-
-
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3
13
2
-
1
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6
-
-
1
-
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2
1
1
2
-
1
1
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1
1
1
1
-
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-
1
-
-
1
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3
-
13
-
2
-
1
-
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1
-
2
1
1
2
-
1
1
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1
1
1
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