BRENDA - Enzyme Database
show all sequences of 1.14.12.13

Purification and some properties of 2-halobenzoate 1,2-dioxygenase, a two-component enzyme system from Pseudomonas cepacia 2CBS

Fetzner, S.; Muller, R.; Lingens, F.; J. Bacteriol. 174, 279-290 (1992)

Data extracted from this reference:

Inhibitors
Inhibitors
Commentary
Organism
Structure
2,2'-dipyridyl
100% inhibition at 1 mM
Burkholderia cepacia
Cu2+
100% inhibition compared to the activity without any metal
Burkholderia cepacia
EDTA
100% inhibition at 1 mM
Burkholderia cepacia
iodoacetate
55% inhibition at 2 mM
Burkholderia cepacia
KCN
40% inhibition at 2 mM
Burkholderia cepacia
additional information
no inhibition with sodium azide
Burkholderia cepacia
N-ethylmaleimide
100% inhibition at 2 mM
Burkholderia cepacia
Ni2+
25% inhibition compared to the activity without any metal
Burkholderia cepacia
o-phenanthroline
100% inhibition at 0.5 mM
Burkholderia cepacia
p-chloromercuribenzoate
17% inhibition at 0.005 mM
Burkholderia cepacia
Zn2+
40% inhibition compared to the activity without any metal
Burkholderia cepacia
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.023
-
2-Chlorobenzoate
component A
Burkholderia cepacia
0.079
-
NADH
component B
Burkholderia cepacia
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Fe2+
exogenous, required for activity
Burkholderia cepacia
Fe2+
component B contains 1.7 mol and A 5.8 mol of iron per mol of enzyme
Burkholderia cepacia
Sulfide
component B contains 1.7 mol and A 6.0 mol of acid-labile sulfide per mol of enzyme
Burkholderia cepacia
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
20000
-
3 * 52000, 3 * 20000, alpha3,beta3 structure of component A, SDS-PAGE
Burkholderia cepacia
37500
-
1 * 37500, component B, SDS-PAGE
Burkholderia cepacia
37500
38000
component B, gel filtration
Burkholderia cepacia
52000
-
3 * 52000, 3 * 20000, alpha3,beta3 structure of component A, SDS-PAGE
Burkholderia cepacia
200000
220000
component A, gel filtration
Burkholderia cepacia
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
2-chlorobenzoate + NADH + O2
Burkholderia cepacia
first step of 2-chlorobenzoate degradation via meta-cleavage
catechol + chloride + NAD+ + CO2
-
Burkholderia cepacia
?
Organism
Organism
UniProt
Commentary
Textmining
Burkholderia cepacia
-
2CBS, isolated from water
-
Purification (Commentary)
Purification (Commentary)
Organism
component A: anion-exchange, gel filtration, component B: anion-exchange, affinity and adsorption chromatography
Burkholderia cepacia
Reaction
Reaction
Commentary
Organism
Reaction ID
a 2-halobenzoate + NADH + H+ + O2 = catechol + a halide anion + NAD+ + CO2
inducible two-component enzyme system: component B is an iron-sulfur flavoprotein (reductase) and red-brown component A (oxygenase) is an iron-sulfur protein
Burkholderia cepacia
a 2-halobenzoate + NADH + H+ + O2 = catechol + a halide anion + NAD+ + CO2
multi-component dioxygenase system proposed: 1,2-hydroxylating, dehalogenating, decarboxylating
Burkholderia cepacia
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
0.0085
-
crude extract, 3.4 mg protein per ml in the assay
Burkholderia cepacia
0.53
-
activity of component A after gel filtration in presence of component B
Burkholderia cepacia
4
-
activity of component B in presence of component A
Burkholderia cepacia
Storage Stability
Storage Stability
Organism
-20°C component A, 80-85% loss of activity
Burkholderia cepacia
-20°C, component A, 20% glycerol, 50% loss of activity
Burkholderia cepacia
-20°C, component B, 20% sucrose, 2 weeks, 25% loss of activity
Burkholderia cepacia
-80°C or -20°C, component B, 2 weeks, 70% loss of activity
Burkholderia cepacia
-80°C or -20°C, component B, 20% glycerol, 2 weeks, 15% loss of activity
Burkholderia cepacia
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
2-aminobenzoate + NADH + O2
-
439074
Burkholderia cepacia
? + NAD+ + CO2
-
-
-
?
2-bromobenzoate + NADH + O2
-
439074
Burkholderia cepacia
catechol + bromide + NAD+ + CO2
-
439074
Burkholderia cepacia
?
2-chlorobenzoate + NADH + O2
-
439074
Burkholderia cepacia
catechol + chloride + NAD+ + CO2
-
439074
Burkholderia cepacia
?
2-chlorobenzoate + NADH + O2
first step of 2-chlorobenzoate degradation via meta-cleavage
439074
Burkholderia cepacia
catechol + chloride + NAD+ + CO2
-
439074
Burkholderia cepacia
?
2-fluorobenzoate + NADH + O2
-
439074
Burkholderia cepacia
catechol + fluoride + NAD+ + CO2
-
439074
Burkholderia cepacia
?
2-hydroxybenzoate + NADH + O2
-
439074
Burkholderia cepacia
catechol + ? + NAD+ + CO2
-
439074
Burkholderia cepacia
?
2-iodobenzoate + NADH + O2
-
439074
Burkholderia cepacia
catechol + iodide + NAD+ + CO2
-
439074
Burkholderia cepacia
?
2-methoxybenzoate + NADH + O2
-
439074
Burkholderia cepacia
catechol + ? + NAD+ + CO2
-
439074
Burkholderia cepacia
?
2-methylbenzoate + NADH + O2
-
439074
Burkholderia cepacia
o-cresol + ? + NAD+ + CO2
-
439074
Burkholderia cepacia
?
anthranilate + NADH + O2
-
439074
Burkholderia cepacia
catechol + ? + NAD+ + CO2
-
439074
Burkholderia cepacia
?
benzoate + NADH + O2
-
439074
Burkholderia cepacia
3,5-cyclohexadiene-1,2-diol-1-carboxylic acid + NAD+ + CO2
-
439074
Burkholderia cepacia
?
additional information
very broad substrate specificity is listed, benzoate analogs with substituents in ortho position are preferred. Trichloroethylene, dihydroxybenzoates, benzene chlorobenzene, toluene, phenol and chlorophenylacetate isomers do not serve as substrates
439074
Burkholderia cepacia
?
-
-
-
?
Subunits
Subunits
Commentary
Organism
hexamer
3 * 52000, 3 * 20000, alpha3,beta3 structure of component A, SDS-PAGE
Burkholderia cepacia
monomer
1 * 37500, component B, SDS-PAGE
Burkholderia cepacia
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
25
-
assay at
Burkholderia cepacia
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6.5
-
assay at
Burkholderia cepacia
Cofactor
Cofactor
Commentary
Organism
Structure
FAD
component B contains 0.8 mol per mol of enzyme, 0.002 mM increases activity
Burkholderia cepacia
FMN
little increase of activity by addition of 0.002 mM
Burkholderia cepacia
NADH
can not be replaced by NADPH
Burkholderia cepacia
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
FAD
component B contains 0.8 mol per mol of enzyme, 0.002 mM increases activity
Burkholderia cepacia
FMN
little increase of activity by addition of 0.002 mM
Burkholderia cepacia
NADH
can not be replaced by NADPH
Burkholderia cepacia
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
2,2'-dipyridyl
100% inhibition at 1 mM
Burkholderia cepacia
Cu2+
100% inhibition compared to the activity without any metal
Burkholderia cepacia
EDTA
100% inhibition at 1 mM
Burkholderia cepacia
iodoacetate
55% inhibition at 2 mM
Burkholderia cepacia
KCN
40% inhibition at 2 mM
Burkholderia cepacia
additional information
no inhibition with sodium azide
Burkholderia cepacia
N-ethylmaleimide
100% inhibition at 2 mM
Burkholderia cepacia
Ni2+
25% inhibition compared to the activity without any metal
Burkholderia cepacia
o-phenanthroline
100% inhibition at 0.5 mM
Burkholderia cepacia
p-chloromercuribenzoate
17% inhibition at 0.005 mM
Burkholderia cepacia
Zn2+
40% inhibition compared to the activity without any metal
Burkholderia cepacia
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.023
-
2-Chlorobenzoate
component A
Burkholderia cepacia
0.079
-
NADH
component B
Burkholderia cepacia
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Fe2+
exogenous, required for activity
Burkholderia cepacia
Fe2+
component B contains 1.7 mol and A 5.8 mol of iron per mol of enzyme
Burkholderia cepacia
Sulfide
component B contains 1.7 mol and A 6.0 mol of acid-labile sulfide per mol of enzyme
Burkholderia cepacia
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
20000
-
3 * 52000, 3 * 20000, alpha3,beta3 structure of component A, SDS-PAGE
Burkholderia cepacia
37500
-
1 * 37500, component B, SDS-PAGE
Burkholderia cepacia
37500
38000
component B, gel filtration
Burkholderia cepacia
52000
-
3 * 52000, 3 * 20000, alpha3,beta3 structure of component A, SDS-PAGE
Burkholderia cepacia
200000
220000
component A, gel filtration
Burkholderia cepacia
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
2-chlorobenzoate + NADH + O2
Burkholderia cepacia
first step of 2-chlorobenzoate degradation via meta-cleavage
catechol + chloride + NAD+ + CO2
-
Burkholderia cepacia
?
Purification (Commentary) (protein specific)
Commentary
Organism
component A: anion-exchange, gel filtration, component B: anion-exchange, affinity and adsorption chromatography
Burkholderia cepacia
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
0.0085
-
crude extract, 3.4 mg protein per ml in the assay
Burkholderia cepacia
0.53
-
activity of component A after gel filtration in presence of component B
Burkholderia cepacia
4
-
activity of component B in presence of component A
Burkholderia cepacia
Storage Stability (protein specific)
Storage Stability
Organism
-20°C component A, 80-85% loss of activity
Burkholderia cepacia
-20°C, component A, 20% glycerol, 50% loss of activity
Burkholderia cepacia
-20°C, component B, 20% sucrose, 2 weeks, 25% loss of activity
Burkholderia cepacia
-80°C or -20°C, component B, 2 weeks, 70% loss of activity
Burkholderia cepacia
-80°C or -20°C, component B, 20% glycerol, 2 weeks, 15% loss of activity
Burkholderia cepacia
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
2-aminobenzoate + NADH + O2
-
439074
Burkholderia cepacia
? + NAD+ + CO2
-
-
-
?
2-bromobenzoate + NADH + O2
-
439074
Burkholderia cepacia
catechol + bromide + NAD+ + CO2
-
439074
Burkholderia cepacia
?
2-chlorobenzoate + NADH + O2
-
439074
Burkholderia cepacia
catechol + chloride + NAD+ + CO2
-
439074
Burkholderia cepacia
?
2-chlorobenzoate + NADH + O2
first step of 2-chlorobenzoate degradation via meta-cleavage
439074
Burkholderia cepacia
catechol + chloride + NAD+ + CO2
-
439074
Burkholderia cepacia
?
2-fluorobenzoate + NADH + O2
-
439074
Burkholderia cepacia
catechol + fluoride + NAD+ + CO2
-
439074
Burkholderia cepacia
?
2-hydroxybenzoate + NADH + O2
-
439074
Burkholderia cepacia
catechol + ? + NAD+ + CO2
-
439074
Burkholderia cepacia
?
2-iodobenzoate + NADH + O2
-
439074
Burkholderia cepacia
catechol + iodide + NAD+ + CO2
-
439074
Burkholderia cepacia
?
2-methoxybenzoate + NADH + O2
-
439074
Burkholderia cepacia
catechol + ? + NAD+ + CO2
-
439074
Burkholderia cepacia
?
2-methylbenzoate + NADH + O2
-
439074
Burkholderia cepacia
o-cresol + ? + NAD+ + CO2
-
439074
Burkholderia cepacia
?
anthranilate + NADH + O2
-
439074
Burkholderia cepacia
catechol + ? + NAD+ + CO2
-
439074
Burkholderia cepacia
?
benzoate + NADH + O2
-
439074
Burkholderia cepacia
3,5-cyclohexadiene-1,2-diol-1-carboxylic acid + NAD+ + CO2
-
439074
Burkholderia cepacia
?
additional information
very broad substrate specificity is listed, benzoate analogs with substituents in ortho position are preferred. Trichloroethylene, dihydroxybenzoates, benzene chlorobenzene, toluene, phenol and chlorophenylacetate isomers do not serve as substrates
439074
Burkholderia cepacia
?
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
hexamer
3 * 52000, 3 * 20000, alpha3,beta3 structure of component A, SDS-PAGE
Burkholderia cepacia
monomer
1 * 37500, component B, SDS-PAGE
Burkholderia cepacia
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
25
-
assay at
Burkholderia cepacia
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6.5
-
assay at
Burkholderia cepacia
Other publictions for EC 1.14.12.13
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
Synonyms
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)
742665
Xu
-
Degradation of three monochlo ...
Pseudomonas sp. 2-CBA
Int. Biodeterior. Biodegrad.
120
192-202
2017
-
-
1
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
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1
-
3
-
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1
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1
1
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1
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1
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-
-
-
-
-
-
-
-
704034
Baggi
-
Co-metabolism of di- and trich ...
Cupriavidus necator, Stenotrophomonas maltophilia
Int. Biodeter. Biodegrad.
62
57-64
2008
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
2
-
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4
-
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2
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-
-
-
-
-
-
2
2
-
-
-
688859
Wittich
Growth of the genetically engi ...
Burkholderia cepacia, Burkholderia cepacia 2CBS
Microbiology
153
186-195
2007
-
-
1
-
-
-
-
-
-
-
-
-
-
7
-
-
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-
-
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2
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1
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-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
439078
Suzuki
Expression of 2-halobenzoate d ...
Burkholderia sp.
Gene
262
137-145
2001
-
-
1
-
1
-
-
-
-
-
-
1
-
10
-
-
-
-
-
-
6
-
6
-
-
1
-
-
-
1
-
-
-
-
-
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-
1
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-
1
-
-
-
-
-
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-
1
-
-
-
-
-
-
6
-
6
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
439079
Francisco
The chlorobenzoate dioxygenase ...
Burkholderia sp., Burkholderia sp. NK8
Microbiology
147
121-133
2001
-
-
1
-
1
-
-
-
-
2
-
2
-
21
-
-
-
1
-
-
3
-
12
-
-
1
-
-
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-
1
-
2
-
-
-
-
-
1
2
-
1
-
-
-
-
-
-
2
-
2
-
-
-
-
-
-
3
-
12
-
1
-
-
-
-
1
-
-
-
-
-
-
-
-
439077
Fava
2-Chlorobenzoic acid and 2,5-d ...
Pseudomonas sp., Pseudomonas sp. CPE2
Lett. Appl. Microbiol.
22
275-279
1996
-
-
-
-
-
-
2
2
-
1
-
4
-
2
-
-
-
-
-
-
2
-
8
-
-
2
-
-
-
2
-
-
2
-
-
-
-
-
-
2
-
-
-
-
2
-
2
-
1
-
4
-
-
-
-
-
-
2
-
8
-
2
-
-
-
2
-
-
-
-
-
-
-
-
-
439075
Haak
Cloning, nucleotide sequence, ...
Burkholderia cepacia
J. Bacteriol.
177
667-675
1995
-
-
1
-
1
-
-
-
-
2
3
1
-
18
-
-
1
1
-
-
-
-
6
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
1
2
-
1
-
-
-
-
-
-
2
3
1
-
-
-
1
-
-
-
-
6
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
439076
Riedel
EPR, electron spin echo envelo ...
Burkholderia cepacia
J. Biol. Chem.
270
30869-30873
1995
-
-
-
-
-
-
-
-
4
2
-
1
-
15
-
-
1
1
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
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-
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-
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-
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4
2
-
1
-
-
-
1
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-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
661983
Romanov
Pseudomonas aeruginosa 142 use ...
Pseudomonas aeruginosa, Pseudomonas aeruginosa 142
J. Bacteriol.
176
3368-3374
1994
-
-
-
-
-
-
1
-
-
1
1
-
-
5
-
-
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-
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-
1
-
20
1
-
-
-
-
-
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-
-
3
-
-
-
-
-
-
3
-
-
-
-
1
-
-
-
1
1
-
-
-
-
-
-
-
1
-
20
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
439074
Fetzner
Purification and some properti ...
Burkholderia cepacia
J. Bacteriol.
174
279-290
1992
-
-
-
-
-
-
11
2
-
3
5
1
-
8
-
-
1
2
-
-
3
5
12
2
-
1
-
-
-
1
-
-
3
-
-
-
-
-
-
3
-
-
-
-
11
-
2
-
3
5
1
-
-
-
1
-
-
3
5
12
2
1
-
-
-
1
-
-
-
-
-
-
-
-
-
439073
Fetzner
Degradation of 2-chlorobenzoat ...
Burkholderia cepacia
Biol. Chem. Hoppe-Seyler
370
1173 - 1182
1989
-
-
-
-
1
1
-
-
-
1
-
1
-
4
-
-
1
1
-
-
2
-
2
-
-
1
-
-
-
1
-
-
1
-
-
-
-
-
-
1
-
1
1
-
-
-
-
-
1
-
1
-
-
-
1
-
-
2
-
2
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-