Information on EC 4.4.1.9 - L-3-cyanoalanine synthase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY hide
4.4.1.9
-
RECOMMENDED NAME
GeneOntology No.
L-3-cyanoalanine synthase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
L-cysteine + hydrogen cyanide = L-3-cyanoalanine + hydrogen sulfide
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
elimination
-
-
beta-replacement; of H2S or RSH, C-S bond cleavage
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Biosynthesis of secondary metabolites
-
-
cyanide degradation
-
-
cyanide detoxification I
-
-
Cyanoamino acid metabolism
-
-
Metabolic pathways
-
-
SYSTEMATIC NAME
IUBMB Comments
L-cysteine hydrogen-sulfide-lyase (adding hydrogen cyanide; L-3-cyanoalanine-forming)
Contains pyridoxal phospate.
CAS REGISTRY NUMBER
COMMENTARY hide
9059-53-4
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
leek
-
-
Manually annotated by BRENDA team
onion
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
cucumber
-
-
Manually annotated by BRENDA team
carnation flower
-
-
Manually annotated by BRENDA team
barnyard grass
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
barley
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Lupinus sp.
-
-
-
Manually annotated by BRENDA team
cv. Fuji
SwissProt
Manually annotated by BRENDA team
apple
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
avocado
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
pea
-
-
Manually annotated by BRENDA team
cv. Micro-Tom
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
southern armyworm
-
-
Manually annotated by BRENDA team
cv. Prelude, gene Tu-CAS
UniProt
Manually annotated by BRENDA team
cabbage looper moth
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
immobilization of enzyme
-
-
Manually annotated by BRENDA team
Xanthium pennsylvanicum
-
-
-
Manually annotated by BRENDA team
maize
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
-
enzyme knockout leads to an increased level of cyanide in the roots and leaves and a severe defect in root hair morphogenesis, suggesting that cyanide acts as a signaling factor in root developmen. The CYS-C1 loss-of-function mutation is not toxic for the plant
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
3-chloro-L-alanine + HCN
?
show the reaction diagram
Lupinus sp.
-
-
-
-
ir
chloroalanine + sulfide
L-cysteine + Cl-
show the reaction diagram
L-cysteine + CN-
sulfide + L-3-cyanoalanine
show the reaction diagram
-
-
-
-
?
L-cysteine + HCN
?
show the reaction diagram
L-cysteine + HCN
H2S + L-3-cyanoalanine
show the reaction diagram
L-cysteine + HCN
sulfide + L-3-cyanoalanine
show the reaction diagram
L-cysteine + hydrogen cyanide
hydrogen sulfide + L-3-cyanoalanine
show the reaction diagram
L-cysteine + methanethiol
S-methyl-L-cysteine + ?
show the reaction diagram
L-cysteine + NaCN
L-3-cyanoalanin + Na2S
show the reaction diagram
L-cysteine + pyrazole
beta-(pyrazol-1-yl)-L-alanine + ?
show the reaction diagram
-
-
-
?
O-acetyl-L-serine + benzylmercaptan
S-benzyl-L-cysteine + acetate
show the reaction diagram
-
-
?
O-acetyl-L-serine + CN-
acetate + L-3-cyanoalanine
show the reaction diagram
-
-
?
O-acetyl-L-serine + ethylmercaptan
S-ethyl-L-cysteine + acetate
show the reaction diagram
-
-
?
O-acetyl-L-serine + methylmercaptan
S-methyl-L-cysteine + acetate
show the reaction diagram
-
-
?
O-acetyl-L-serine + phenol
S-phenyl-L-serine + acetate
show the reaction diagram
-
-
?
O-acetyl-L-serine + phenylmercaptan
S-phenyl-L-cysteine + acetate
show the reaction diagram
-
-
?
O-acetyl-L-serine + propylmercaptan
S-propyl-L-cysteine + acetate
show the reaction diagram
-
-
?
O-acetyl-L-serine + sulfide
L-cysteine + acetate
show the reaction diagram
O-acetylserine + H2S
cysteine + acetic acid
show the reaction diagram
O-acetylserine + HCN
beta-cyanoalanine + acetic acid
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
chloroalanine + sulfide
L-cysteine + Cl-
show the reaction diagram
-
-
-
?
L-cysteine + HCN
?
show the reaction diagram
L-cysteine + HCN
H2S + L-3-cyanoalanine
show the reaction diagram
-
-
-
-
ir
L-cysteine + HCN
sulfide + L-3-cyanoalanine
show the reaction diagram
L-cysteine + hydrogen cyanide
hydrogen sulfide + L-3-cyanoalanine
show the reaction diagram
O-acetyl-L-serine + CN-
acetate + L-3-cyanoalanine
show the reaction diagram
Q84IF9
-
-
?
O-acetyl-L-serine + sulfide
L-cysteine + acetate
show the reaction diagram
additional information
?
-
T1KF23
the enzyme is also active with O-acetyl-L-serine, EC 2.5.1.47, but the L-3-cyanoalanine forming activity is highly preferred
-
-
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
pyridoxal 5'-phosphate
additional information
Lupinus sp.
-
no cofactors required
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
slight stimulation
Zn2+
-
slight stimulation
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3-aminopropionate
Lupinus sp.
-
-
aminooxyacetic acid
Benzoic acid
-
-
Beta-cyano-L-alanine
-
-
chloroalanine
-
uncompetitive vs. L-cysteine
chloroanaline
-
substrate inhibition
-
cyanide
-
-
cyanoalanine
-
uncompetitive vs. O-acetyl-L-serine, noncompetitive vs. chloroalanine and L-cysteine
ethylene
-
down-regulates CAS expression in etiolated rice seedlings
HgCl2
Lupinus sp.
-
-
homocysteine
-
uncompetitive vs. O-acetyl-L-serine, noncompetitive vs. chloroalanine and L-cysteine
hydroxylamine
iodoacetamide
iodoacetic acid
L-cysteine
L-methionine
-
-
N-ethylmaleimide
Lupinus sp.
-
-
NaBH4
O-acetyl-L-serine
p-chloromercuribenzoate
salicylic acid
-
-
Semicarbazide
-
slight
Sodium borohydride
-
-
sodium thiocyanate
-
uncompetitive vs. O-acetyl-L-serine
sulfhydryl reagents
-
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1-amino-cyclopropane-1-carboxylate
-
induces CAS expression
1-aminocyclopropane-1-carboxylic acid
-
stimulates
2,4-D
-
activates CAS expression at 10 mM, 2,4-D is a synthetic auxin
3-allyloxy-1,2-benzisothiazole-1,1-dioxide
-
co-induction by cell wall protein with elicitin activity from Pythium oligandrum and 3-allyloxy-1,2-benzisothiazole-1,1-dioxide
auxin
-
upregulates CAS expression
benzo(1,2,3)thiadiazole-7-carbothioic acid S-methyl ester
-
-
brassinolide
-
increases CAS activity at 0.01 mM
ethylene
L-asparagine
-
-
L-cysteine
-
-
quinclorac
-
increases CAS activity at 0.1 mM
validamycin A
-
-
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.66 - 10
2-mercaptoethanol
0.54
chloroalanine
-
-
0.6
chloroanaline
-
26C, pH 9.8
-
0.13 - 0.235
CN-
0.5 - 5.5
cyanide
0.87
cysteine
Lupinus sp.
-
-
0.27 - 0.67
HCN
0.26
hydrogen cyanide
recombinant enzyme, pH 9.0, 25C
0.02 - 55
KCN
0.14 - 3.6
L-cysteine
0.04 - 8.24
Na2S
0.73 - 6.25
NaCN
0.134 - 39.88
O-acetyl-L-serine
9.51
Sulfide
-
pH 8.5
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
39.2
hydrogen cyanide
Glycine max
I1L6I6
recombinant enzyme, pH 9.0, 25C
2.17 - 2.94
KCN
2.67 - 2135
L-cysteine
1.5
Na2S
Arabidopsis thaliana
-
30C, pH 8.0
2
O-acetyl-L-serine
Arabidopsis thaliana
-
30C, pH 8.0
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
150.8
hydrogen cyanide
Glycine max
I1L6I6
recombinant enzyme, pH 9.0, 25C
20302
48.02
L-cysteine
Glycine max
I1L6I6
recombinant enzyme, pH 9.0, 25C
74
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.27
chloroalanine
-
vs. L-cysteine
6.9
chloroanaline
-
26C, pH 9.8
-
0.86 - 30
cyanoalanine
1.29 - 10.1
homocysteine
0.87 - 2.59
KCN
5.22
L-cysteine
-
26C, pH 9.8
5 - 38.1
O-acetyl-L-serine
1.45 - 15.1
sodium thiocyanate
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
20 - 40
cyanide
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.00034
-
latex pellet
0.00052
-
inner bark
0.003
-
immobilized enzyme
0.0043
-
latex cytosol
0.0375
-
purified enzyme
0.25
substrate L-cysteine
0.43
substrate O-acetyl-L-serine
6.84
pH 8.5, 37C, purified recombinant His-tagged enzyme
13.3
-
wild type enzyme, after 750fold purification
33.8
Lupinus sp.
-
-
38
Xanthium pennsylvanicum
-
-
43.5
Lupinus sp.
-
-
49.5
substrate O-acetyl-L-serine
62.1
-
substrate L-cysteine
66.2
substrate L-cysteine
111.3
-
recombinant enzyme from Saccharomyces cerevisiae, after 750fold purification
157
substrate L-cysteine
181
-
recombinant enzyme from Arabidopsis thaliana, after 750fold purification
323.8
Lupinus sp.
-
-
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 9.5
-
dissolved
8.5 - 9.2
-
-
8.5 - 9.5
-
-
8.8 - 9.5
Lupinus sp.
-
-
8.9
-
assay at
9 - 9.5
-
-
9
-
activity in borate buffer
10.5
Xanthium pennsylvanicum
-
cytosolic enzyme
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5 - 10.5
approx. 50% of maximal activity at pH 4.5 and pH 10.5, respectively
7.5 - 13
-
-
7.5 - 11
Lupinus sp.
-
-
10 - 11.5
-
sharp decrease in activity below pH 10.0, 15% of maximal activity at pH 11.5
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
assay at
35
-
assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25 - 40
-
-
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.7
-
isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
; activity in the rubber tree latex cytosol is almost 10fold higher than in the bark
Manually annotated by BRENDA team
6.2 times and 2.5 times higher levels than in tubers and leaves, respectively
Manually annotated by BRENDA team
Lupinus sp.
-
-
Manually annotated by BRENDA team
-
activity in the rubber tree latex cytosol is almost 10fold higher than in the bark; cytosol, pellet
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
additional information
-
not: vacuole and chloroplast fractions
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
23250
-
2 * 23250, SDS-PAGE
28000
-
2 * 28000, SDS-PAGE
30000
-
2 * 30000, SDS-PAGE
33500
-
2 * 33500, SDS-PAGE
34000
2 * 34000, SDS-PAGE
35000
-
2 * 35000, SDS-PAGE
37000
-
2 * 37000, SDS-PAGE
37600
x * 37600, immunoblot
38200
x * 38200, immunoblot
40000
-
2 * 40000, SDS-PAGE
40500
sequence analysis
40900
sequence analysis
52000 - 53000
Lupinus sp.
-
gel filtration
56000
-
gel filtration
60000 - 70000
-
gel filtration
60000
-
gel filtration
552300
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 37600, immunoblot; x * 38200, immunoblot
homodimer
monomer
Lupinus sp.
-
-
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant wild-type and K95A mutant enzymes, hanging drops vapor diffusion method , 0004 ml of 10-12 mg/ml protein solution is mixed with an equal volume of crystallization solution containing 10% PEG 8000, 0.2 M NaCl, and 0.1 sodium/potassium phosphate, pH 7.0, and 0.2 M MgSO4, at 4C over a 0.7 mL reservoir. The K95A mutant is crystallized in similar drops from a solution containing 20% PEG 3350, 0.2 M potassium citrate, pH 7.5, X-ray diffraction structure determination and analysis at 2.50 A and 1.77 A resolution, respectively, molecular replacement
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8 - 8.5
-
highest stability
34722
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
60
stable for 30 min at pH 6.0-10.0
70
no loss of activity after 30 min at pH 7.5
additional information
-
activity reduces to near zero at 45C
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
immobilized enzyme, 6C, 10% loss of activity after 70 days
-
stable at -20C
Lupinus sp.
-
stable for more than 1 month at -20C
Lupinus sp.
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
80% acetone, ammonium sulfate, DEAE Sephacel, Mono Q, Resource PHE, preparative electrophoresis, TSK gel 3000
-
ammonium sulfate, ethanol, heat, DEAE-Cellulofine, gel filtration, Phenyl-Sepharose
anti-GFP microbead immunopurification and Mono Q colum chromatography
-
by ion-exchange chromatography and gel filtration
-
recombinant AtcysC1
-
recombinant functional N-terminally His-tagged wild-type and mutant enzymes lacking the mitochondrial localization sequence by nickel affinity chromatography and gel filtration
recombinant PCAS-1; recombinant PCAS-2
recombinant soluble N-terminally His-tagged enzyme from Escherichia coli by nickel affinity chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Arabidopsis thaliana and in Saccharomyces cerevisiae
-
expression in Escherichia coli
expression in Escherichia coli; expression in Escherichia coli
expression of PCAS-1 in Escherichia coli; expression of PCAS-2 in Escherichia coli
full-length MdCAS2 cDNA clones introduced into plasmid pMAL-c2X, expressed in Escherichia coli BL21 (DE3); full-length pMdCAS1 cDNA clones introduced into plasmid pMAL-c2X, expressed in Escherichia coli BL21 (DE3)
gene Tu-CAS , DNA and amino acid sequence determination and analysis, phylogenetic analysis, recombinant expression of N-terminally His-tagged enzyme in Escherichia coli
recombinant enzyme expression as N-terminally His-tagged, functional dimeric type and mutant enzyme proteins lacking the mitochondrial localization sequence
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
K95A
site-directed mutagenesis, inactive mutant
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
agriculture