BRENDA - Enzyme Database
show all sequences of 4.1.2.44

Aerobic benzoyl-CoA catabolic pathway in Azoarcus evansii: studies on the non-oxygenolytic ring cleavage enzyme

Gescher, J.; Eisenreich, W.; Wörth, J.; Bacher, A.; Fuchs, G.; Mol. Microbiol. 56, 1586-1600 (2005)

Data extracted from this reference:

Activating Compound
Activating Compound
Commentary
Organism
Structure
additional information
addition of 1 mM thiamine diphosphate to the standard assay causes only a minimal stimulation (8%)
Azoarcus evansii
Cloned(Commentary)
Commentary
Organism
the boxC gene is expressed in a recombinant Escherichia coli strain as a fusion protein with maltose binding protein (BoxCmal)
Azoarcus evansii
Inhibitors
Inhibitors
Commentary
Organism
Structure
additional information
acetoacetyl-CoA (0.2 mM), a potential inhibitor of enoyl-CoA hydratase, has no impact on enzyme activity of BoxCmal. Crotonyl-CoA (0.2 mM), a potential substrate of enoyl-CoA hydratase (crotonase), is neither converted to 3-hydroxybutyryl-CoA nor does it inhibit the standard assay
Azoarcus evansii
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.017
-
2,3-dihydro-2,3-dihydroxybenzoyl-CoA
-
Azoarcus evansii
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
additional information
the purified protein does not require divalent metals or any cosubstrates or coenzymes for activity
Azoarcus evansii
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
60000
-
2 * 60000, SDS-PAGE
Azoarcus evansii
120000
-
gel filtration
Azoarcus evansii
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2,3-dihydro-2,3-dihydroxybenzoyl-CoA + H2O
Azoarcus evansii
the enzyme is involved in the aerobic benzoyl-CoA catabolic pathway. Benzoyl-CoA is oxidized to 2,3-dihydro-2,3-dihydroxybenzoyl-CoA (benzoyl-CoA dihydrodiol) by benzoyl-CoA oxygenase/reductase BoxBA in the presence of molecular oxygen. The next, ring cleaving step is catalysed by BoxC
3,4-dehydroadipyl-CoA semialdehyde + formate
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Azoarcus evansii
Q84HH6
-
-
Purification (Commentary)
Commentary
Organism
wild type and recombinant proteins
Azoarcus evansii
Source Tissue
Source Tissue
Commentary
Organism
Textmining
culture condition:benzoate-grown cell
-
Azoarcus evansii
-
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
4.9
-
-
Azoarcus evansii
Storage Stability
Storage Stability
Organism
-20°C, the protein can be stored without appreciable loss of activity for months in the presence of 10% (v/v) glycerol
Azoarcus evansii
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2,3-dihydro-2,3-dihydroxybenzoyl-CoA + H2O
the enzyme is involved in the aerobic benzoyl-CoA catabolic pathway. Benzoyl-CoA is oxidized to 2,3-dihydro-2,3-dihydroxybenzoyl-CoA (benzoyl-CoA dihydrodiol) by benzoyl-CoA oxygenase/reductase BoxBA in the presence of molecular oxygen. The next, ring cleaving step is catalysed by BoxC
700259
Azoarcus evansii
3,4-dehydroadipyl-CoA semialdehyde + formate
-
-
-
?
2,3-dihydro-2,3-dihydroxybenzoyl-CoA + H2O
NADPH and semicarbazide are analysed directly by NMR spectroscopy and mass spectrometry. The purified protein does not require molecular oxygen for activity
700259
Azoarcus evansii
3,4-dehydroadipyl-CoA semialdehyde + formate
-
-
-
?
additional information
no activity with crotonyl-CoA
700259
Azoarcus evansii
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
homodimer
2 * 60000, SDS-PAGE
Azoarcus evansii
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
20
-
2,3-dihydro-2,3-dihydroxybenzoyl-CoA
-
Azoarcus evansii
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
9
-
-
Azoarcus evansii
pH Range
pH Minimum
pH Maximum
Commentary
Organism
7
11
half maximal activity at pH 7 and pH 11
Azoarcus evansii
Cofactor
Cofactor
Commentary
Organism
Structure
additional information
the purified protein does not require divalent metals or any cosubstrates or coenzymes for activity
Azoarcus evansii
pI Value
Organism
Commentary
pI Value Maximum
pI Value
Azoarcus evansii
-
-
5.6
Activating Compound (protein specific)
Activating Compound
Commentary
Organism
Structure
additional information
addition of 1 mM thiamine diphosphate to the standard assay causes only a minimal stimulation (8%)
Azoarcus evansii
Cloned(Commentary) (protein specific)
Commentary
Organism
the boxC gene is expressed in a recombinant Escherichia coli strain as a fusion protein with maltose binding protein (BoxCmal)
Azoarcus evansii
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
additional information
the purified protein does not require divalent metals or any cosubstrates or coenzymes for activity
Azoarcus evansii
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
additional information
acetoacetyl-CoA (0.2 mM), a potential inhibitor of enoyl-CoA hydratase, has no impact on enzyme activity of BoxCmal. Crotonyl-CoA (0.2 mM), a potential substrate of enoyl-CoA hydratase (crotonase), is neither converted to 3-hydroxybutyryl-CoA nor does it inhibit the standard assay
Azoarcus evansii
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.017
-
2,3-dihydro-2,3-dihydroxybenzoyl-CoA
-
Azoarcus evansii
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
additional information
the purified protein does not require divalent metals or any cosubstrates or coenzymes for activity
Azoarcus evansii
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
60000
-
2 * 60000, SDS-PAGE
Azoarcus evansii
120000
-
gel filtration
Azoarcus evansii
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2,3-dihydro-2,3-dihydroxybenzoyl-CoA + H2O
Azoarcus evansii
the enzyme is involved in the aerobic benzoyl-CoA catabolic pathway. Benzoyl-CoA is oxidized to 2,3-dihydro-2,3-dihydroxybenzoyl-CoA (benzoyl-CoA dihydrodiol) by benzoyl-CoA oxygenase/reductase BoxBA in the presence of molecular oxygen. The next, ring cleaving step is catalysed by BoxC
3,4-dehydroadipyl-CoA semialdehyde + formate
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
wild type and recombinant proteins
Azoarcus evansii
Source Tissue (protein specific)
Source Tissue
Commentary
Organism
Textmining
culture condition:benzoate-grown cell
-
Azoarcus evansii
-
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
4.9
-
-
Azoarcus evansii
Storage Stability (protein specific)
Storage Stability
Organism
-20°C, the protein can be stored without appreciable loss of activity for months in the presence of 10% (v/v) glycerol
Azoarcus evansii
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2,3-dihydro-2,3-dihydroxybenzoyl-CoA + H2O
the enzyme is involved in the aerobic benzoyl-CoA catabolic pathway. Benzoyl-CoA is oxidized to 2,3-dihydro-2,3-dihydroxybenzoyl-CoA (benzoyl-CoA dihydrodiol) by benzoyl-CoA oxygenase/reductase BoxBA in the presence of molecular oxygen. The next, ring cleaving step is catalysed by BoxC
700259
Azoarcus evansii
3,4-dehydroadipyl-CoA semialdehyde + formate
-
-
-
?
2,3-dihydro-2,3-dihydroxybenzoyl-CoA + H2O
NADPH and semicarbazide are analysed directly by NMR spectroscopy and mass spectrometry. The purified protein does not require molecular oxygen for activity
700259
Azoarcus evansii
3,4-dehydroadipyl-CoA semialdehyde + formate
-
-
-
?
additional information
no activity with crotonyl-CoA
700259
Azoarcus evansii
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
homodimer
2 * 60000, SDS-PAGE
Azoarcus evansii
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
20
-
2,3-dihydro-2,3-dihydroxybenzoyl-CoA
-
Azoarcus evansii
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
9
-
-
Azoarcus evansii
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
7
11
half maximal activity at pH 7 and pH 11
Azoarcus evansii
pI Value (protein specific)
Organism
Commentary
pI Value Maximum
pI Value
Azoarcus evansii
-
-
5.6
Other publictions for EC 4.1.2.44
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)
712441
Rather
Coenzyme A-dependent aerobic m ...
Azoarcus evansii, Azoarcus evansii KB740 (DSM Z6869)
J. Biol. Chem.
285
20615-20624
2010
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-
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2
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2
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-
-
-
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-
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2
-
-
-
-
-
-
-
-
-
-
-
-
-
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-
700259
Gescher
Aerobic benzoyl-CoA catabolic ...
Azoarcus evansii
Mol. Microbiol.
56
1586-1600
2005
1
-
1
-
-
-
1
1
-
1
2
1
-
4
-
-
1
-
-
1
1
1
3
1
-
-
-
1
1
1
-
1
-
1
-
1
-
1
1
-
-
-
-
1
-
1
-
1
2
1
-
-
-
1
-
1
1
1
3
1
-
-
-
1
1
1
-
1
-
-
-
-
-
-
698566
Gescher
Genes coding for a new pathway ...
Azoarcus evansii
J. Bacteriol.
184
6301-6315
2002
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1
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1
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1
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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
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1
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-
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-
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1
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-
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1
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