Information on EC 3.5.5.8 - thiocyanate hydrolase

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The expected taxonomic range for this enzyme is: Proteobacteria

EC NUMBER
COMMENTARY
3.5.5.8
-
RECOMMENDED NAME
GeneOntology No.
thiocyanate hydrolase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
thiocyanate + 2 H2O = carbonyl sulfide + NH3 + HO-
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of thiocyanate
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
thiocyanate degradation II
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SYSTEMATIC NAME
IUBMB Comments
thiocyanate aminohydrolase
The enzyme from Thiobacillus thioparus catalyses the first step in the degradation of thiocyanate.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SCNase
O66186, O66187, O66188
-
SCNase
Thiobacillus thioparus THI115
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
142539-65-9
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
subunit alpha
UniProt
Manually annotated by BRENDA team
subunit beta
UniProt
Manually annotated by BRENDA team
subunit gamma
UniProt
Manually annotated by BRENDA team
Thiobacillus thioparus THI115
strain THI115
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
thiocyanate + H2O
carbonyl sulfide + NH3 + OH-
show the reaction diagram
-
-
-
-
?
thiocyanate + H2O
carbonyl sulfide + NH3 + OH-
show the reaction diagram
-
-
-
-
thiocyanate + H2O
carbonyl sulfide + NH3 + OH-
show the reaction diagram
-
-
-
-
thiocyanate + H2O
carbonyl sulfide + NH3 + OH-
show the reaction diagram
-
-
-
?
thiocyanate + H2O
carbonyl sulfide + NH3 + OH-
show the reaction diagram
-
-
-
-
?
thiocyanate + H2O
carbonyl sulfide + NH3 + OH-
show the reaction diagram
-
-
-
-
?
thiocyanate + H2O
carbonyl sulfide + NH3 + OH-
show the reaction diagram
-
-
-
-
?
thiocyanate + H2O
carbonyl sulfide + NH3 + OH-
show the reaction diagram
O66186, O66187, O66188
-
-
-
?
thiocyanate + H2O
carbonyl sulfide + NH3 + OH-
show the reaction diagram
-
via multiple reaction steps
-
-
?
thiocyanate + H2O
carbonyl sulfide + NH3 + OH-
show the reaction diagram
Thiobacillus thioparus THI115
-
-
-
-
?
thiocyanate + H2O
carbonyl sulfide + NH3 + OH-
show the reaction diagram
Thiobacillus thioparus THI115
-
via multiple reaction steps
-
-
?
thiocyanate + H2O
carbonyl sulfide + NH3 + OH-
show the reaction diagram
Thiobacillus thioparus THI115
-
-
-
-
?
additional information
?
-
-
does not use thiosulfate as a substrate
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
thiocyanate + H2O
carbonyl sulfide + NH3 + OH-
show the reaction diagram
-
-
-
-
thiocyanate + H2O
carbonyl sulfide + NH3 + OH-
show the reaction diagram
-
-
-
-
thiocyanate + H2O
carbonyl sulfide + NH3 + OH-
show the reaction diagram
-
-
-
?
thiocyanate + H2O
carbonyl sulfide + NH3 + OH-
show the reaction diagram
-
-
-
-
?
thiocyanate + H2O
carbonyl sulfide + NH3 + OH-
show the reaction diagram
Thiobacillus thioparus THI115
-
-
-
-
?
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Co2+
-
cobalt-containing metalloenzyme, 0.86 Co2+ per enzyme trimer, binding to the gamma-subunit requires the presence of activator protein P15K mediating the Co2+ incorporation, overview
Co2+
-
cobalt-containing metalloenzyme, 1 Co2+ per enzyme trimer, binding site, H124VVVCTLCSCYPRPILGQSPEWYR147, in the gamma-subunit with close relation to nitrile hydratase, absorption spectrum analysis, overview
Co2+
-
contains 0.64 atoms Co2+ per heterotrimer
Co3+
-
metalloenzyme, high active-site sequence homology with nitrile hydratase suggests ligation of Co3+ in metal center similar to nitrile hydratase
Co3+
-
contains a five-coordinate cobalt(III) center
Fe2+
-
contains 0.41 atoms Fe2+ per heterotrimer
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
acetate
-
inhibitory at 20 mM
azide
-
inhibitory at 20 mM
Borate
-
inhibitory at 20 mM
nitrate
-
inhibitory at 20 mM
-
nitrite
-
inhibitory at 20 mM
sulfite
-
inhibitory at 20 mM
Cyanate
-
significant inhibition
additional information
-
significant inhibition (97%) in the activity of thiocyanate hydrolase is observed when the enzyme is bubbled with CO
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ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
additional information
-
expression of thiocyanate hydrolase is promoted by its activator protein P15K
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
11
-
Thiocyanate
-
-
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.03
-
-
crude extract, in 0.1 M potassium phosphate buffer at 37C and pH 7.5
3.3
-
-
after 110fold purification, in 0.1 M potassium phosphate buffer at 37C and pH 7.5
32
-
-
purified native enzyme
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
30
40
-
enzyme shows high activity within this range
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
additional information
-
the organism is obligately chemilithoautotrophic sulfur-oxidizing and uses thiocyanate as sole energy source
Manually annotated by BRENDA team
additional information
Thiobacillus thioparus THI115
-
the organism is obligately chemilithoautotrophic sulfur-oxidizing and uses thiocyanate as sole energy source
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
126000
-
-
gel filtration
130000
-
-
gel filtration
140000
-
-
native and apo-enzyme, gel filtration
142000
-
-
native PAGE
240000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
dodecamer
-
heterododecamer (alphabetagamma)4 of heterotrimers
dodecamer
Thiobacillus thioparus THI115
-
heterododecamer (alphabetagamma)4 of heterotrimers
-
heterododecamer
-
x-ray crystallography
heterododecamer
Thiobacillus thioparus THI115
-
x-ray crystallography
-
heterohexamer
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2 * 29000 + 2 * 19000 + 2 * 17000, gel filtration
hexamer
-
2 * 19000 + 2 * 23000 + 2 * 32000, deduced from amino acid sequence, SDS-PAGE
trimer
-
heterotrimer alphabetagamma
trimer
Thiobacillus thioparus THI115
-
heterotrimer alphabetagamma
-
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
Cys113 is posttranslationally oxidized to cysteinesulfinic acid
additional information
Thiobacillus thioparus THI115
-
Cys113 is posttranslationally oxidized to cysteinesulfinic acid
-
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
hanging drop vapor diffusion method, using 0.1 M potassium phosphate (pH 7.2) containing 2.0 M ammonium sulfate for purified SCNase or 0.1 M potassium phosphate (pH 6.9) containing 1.5 M Na/K tartrate for spontaneously activated SCNase and inactivated SCNase, respectively; hanging drop vapor diffusion method, using 0.1 M potassium phosphate (pH 7.2) containing 2.0 M ammonium sulfate for purified SCNase or 0.1 M potassium phosphate (pH 6.9) containing 1.5 M Na/K tartrate for spontaneously activated SCNase and inactivated SCNase, respectively; hanging drop vapor diffusion method, using 0.1 M potassium phosphate (pH 7.2) containing 2.0 M ammonium sulfate for purified SCNase or 0.1 M potassium phosphate (pH 6.9) containing 1.5 M Na/K tartrate for spontaneously activated SCNase and inactivated SCNase, respectively
O66186, O66187, O66188
hanging drop vapour diffusion method with 0.1 M potassium phosphate (pH 7.6) containing 1.5 M Na/K L-(+) tartrate or 0.1 M potassium phosphate (pH 7.2) containing 2 M ammonium sulfate for apo- or native SCNase, respectively
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TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
60
-
-
treatment of the enzyme at 60C for 5 min decreases activity below 10% of the original
OXIDATION STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
when recombinant SCNase, containing gammaCys131-SO2 and gammaCys133-SO(H) modifications in the cobalt center, is stored at 20C under aerobic conditions for 4 months, no enzymatic activity is detected
-
712237, 712237
when recombinant SCNase, containing gammaCys131-SO2- and gammaCys133-SO(H) modifications in the cobalt center, is stored at 20C under aerobic conditions for 4 months, no enzymatic activity is detected
O66186, O66187, O66188
712237
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
ammonium sulfate precipitation, DEAE-Sephacyl, hydroxylapatite
-
native enzyme from strain THI115, and recombinant enzyme from Echerichia coli strain BL21(DE3) by three steps of butyl-resin chromatography, hydroxyapatite and ion exchange chromatography, or by 4 steps of butyl-resin chromatography and gel filtration
-
DEAE-Sepharose column chromatography, Mono Q GI column chromatography, and Superdex 200 gel filtration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
DNA and amino acid sequence determination and analysis
-
expressed in Escherichia coli BL21(DE3) cells; expressed in Escherichia coli BL21(DE3) cells; expressed in Escherichia coli BL21(DE3) cells
O66186, O66187, O66188
expression in Escherichia coli
-
expression of the alpha, beta, and gamma subunits in Escherichia coli, the subunits assemble to an apo-heterododecamer (alphabetagamma)4 like the wild-type enzyme exhibiting no catalytic activity due to lack of bound Co2+ irrespective of the cobalt concentration in the medium, coepression of enzyme with activator protein P15K leads to a functional Co2+-bound enzyme which exhibits 78% of native enzyme activity, optimization of conditions, overview
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