Information on EC 4.4.1.15 - D-cysteine desulfhydrase

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

EC NUMBER
COMMENTARY
4.4.1.15
-
RECOMMENDED NAME
GeneOntology No.
D-cysteine desulfhydrase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
D-cysteine + H2O = sulfide + NH3 + pyruvate
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
C-S-bond cleavage
-
-
-
-
C-S-bond cleavage
-
-
C-S-bond cleavage
-
-
C-S-bond cleavage
B2MWN0
-
C-S-bond cleavage
Pseudomonas putida UW4, Synechococcus elongatus PCC7942
-
-
-
elimination of H2S or RSH
-
-
-
-
elimination of H2S or RSH
-
-
elimination of H2S or RSH
Synechococcus elongatus PCC7942
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Cysteine and methionine metabolism
-
SYSTEMATIC NAME
IUBMB Comments
D-cysteine sulfide-lyase (deaminating; pyruvate-forming)
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
cysteine desulfhydrase
-
CD
cysteine desulfhydrase
Synechococcus elongatus PCC7942
-
CD
-
D-cysteine desulfhydrase
-
-
D-cysteine desulfhydrase
-
-
D-cysteine desulfhydrase
-
-
D-cysteine desulfhydrase
Pseudomonas putida UW4
-
-
-
D-cysteine desulfhydrase
B2MWN0
-
D-cysteine desulfhydrase
-
-
D-cysteine desulfhydrase
Synechococcus elongatus PCC7942
-
-
-
D-cysteine lyase
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
84012-74-8
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
green alga, strain 211-8b, formerly Chlorella pyrenoidosa strain 211-8b
-
-
Manually annotated by BRENDA team
ecotype C24
-
-
Manually annotated by BRENDA team
cv. small sugar pumpkin
-
-
Manually annotated by BRENDA team
strain W3110 deltatrpED102/F'deltatrpED102
-
-
Manually annotated by BRENDA team
various strains
-
-
Manually annotated by BRENDA team
Escherichia coli W3110
strain W3110 deltatrpED102/F'deltatrpED102
-
-
Manually annotated by BRENDA team
tobacco, var. Samsun
-
-
Manually annotated by BRENDA team
strain CU 1462/7
-
-
Manually annotated by BRENDA team
Phormidium uncinatum CU 1462/7
strain CU 1462/7
-
-
Manually annotated by BRENDA team
strain CR 1-1
-
-
Manually annotated by BRENDA team
strain UW4
-
-
Manually annotated by BRENDA team
Pseudomonas putida CR 1-1
strain CR 1-1
-
-
Manually annotated by BRENDA team
Pseudomonas putida UW4
strain UW4
-
-
Manually annotated by BRENDA team
bony best variety
UniProt
Manually annotated by BRENDA team
strain PCC7942
-
-
Manually annotated by BRENDA team
Synechococcus elongatus PCC7942
strain PCC7942
-
-
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
3-chloro-D-alanine + H2O
pyruvate + NH3 + Cl-
show the reaction diagram
-
-
-
-
?
3-chloro-D-alanine + H2O
pyruvate + NH3 + Cl-
show the reaction diagram
-
alpha,beta-elimination, more effective substrate than D-cysteine
-
-
-
3-chloro-D-alanine + H2O
pyruvate + NH3 + Cl-
show the reaction diagram
-
alpha,beta-elimination, more effective substrate than D-cysteine
-
?
3-chloro-D-alanine + H2O
pyruvate + NH3 + Cl-
show the reaction diagram
Pseudomonas putida CR 1-1, Escherichia coli W3110
-
-
-
-
?
3-chloro-D-alanine + H2O
pyruvate + NH3 + Cl-
show the reaction diagram
Escherichia coli W3110
-
alpha,beta-elimination, more effective substrate than D-cysteine
-
?
3-chloro-D-alanine + thioglycolic acid
S-carboxymethyl-D-cysteine
show the reaction diagram
Escherichia coli, Escherichia coli W3110
-
i.e. mercaptoacetic acid, beta-replacement reaction
product is further metabolized to form pyruvate
?
D-allocystathionine + H2O
?
show the reaction diagram
-
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
B2MWN0
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
additional product suggested
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
more sulfide than pyruvate and ammonium formed
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
more sulfide than pyruvate and ammonium formed
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
alpha,beta-elimination
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
very specific, no activity with L-cysteine, mercaptoacetic acid, mercaptoethanol, D-cystine, L-cystine, cysteamine, L-cysteine methylester and dithioerythritol
additional product suggested, more sulfide than pyruvate and ammonium formed
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
very specific, only traces of activity with mercaptoacetic acid, mercaptoethanol, D-cystine, L-cystine, cysteamine, L-cysteine methylester
additional product suggested, more sulfide than pyruvate and ammonium formed
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
plant sulfur metabolism
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
plant sulfur metabolism
additional product suggested, more sulfide than pyruvate and ammonium formed
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
physiological function unknown, detoxification of D-cysteine suggested, contributes to utilization of D-cysteine as sulfur source
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
37C, pH 8.0
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
Pseudomonas putida CR 1-1
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
Pseudomonas putida UW4
-
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
Synechococcus elongatus PCC7942
-
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
Escherichia coli W3110
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
Escherichia coli W3110
-
-, alpha,beta-elimination
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
Phormidium uncinatum CU 1462/7
-
-
-
?
D-cystine + H2O
?
show the reaction diagram
-
-
-
-
?
DL-lanthionine + H2O
?
show the reaction diagram
-
-
-
-
?
DL-selenocysteine + H2O
selenide + L-alanine
show the reaction diagram
-
very poor substrate
-
-
?
DL-selenocystine + H2O
?
show the reaction diagram
-
-
-
-
?
L-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
-
-
?
L-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
5% activity compared to D-cysteine
-
-
?
O-acetyl-D-serine + H2O
?
show the reaction diagram
Escherichia coli, Escherichia coli W3110
-
alpha,beta-elimination reaction
-
-
?
O-acetyl-D-serine + sulfide
D-cysteine + ?
show the reaction diagram
Escherichia coli, Escherichia coli W3110
-
beta-replacement reaction
-
?
O-methyl-DL-serine + H2O
?
show the reaction diagram
-
poor substrate
-
-
?
S-benzyl-D-cysteine + H2O
benzyl hydrosulfide + pyruvate + NH3
show the reaction diagram
-
-
-
-
?
S-carboxymethyl-D-cysteine + H2O
mercaptoacetic acid + pyruvate + NH3
show the reaction diagram
-
-
-
-
?
S-ethyl-D-cysteine + H2O
ethanethiol + pyruvate + NH3
show the reaction diagram
-
-
-
-
?
S-methyl-D-cysteine + H2O
methanethiol + pyruvate + NH3
show the reaction diagram
-
-
-
-
?
S-n-butyl-D-cysteine + H2O
butane-1-thiol + pyruvate + NH3
show the reaction diagram
-
-
-
-
?
S-n-propyl-D-cysteine + H2O
propane-1-thiol + pyruvate + NH3
show the reaction diagram
-
-
-
-
?
S-phenyl-D-cysteine + H2O
benzenthiol + pyruvate + NH3
show the reaction diagram
-
-
-
-
?
L-selenocystine + H2O
selenide + L-alanine
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
L-cysteine is not utilized
-
-
-
additional information
?
-
-
L-cysteine is not utilized
-
-
-
additional information
?
-
-
L-cysteine is not utilized
-
-
-
additional information
?
-
-
L-cysteine is not utilized
-
-
-
additional information
?
-
-
L-cysteine is not utilized
-
-
-
additional information
?
-
-
extracellular production of hydrogen selenide accounts for thiol-assisted toxicity of selenite against Saccharomyces cerevisiae
-
-
-
additional information
?
-
B2MWN0
Site-directed mutagenesis shows that altering only two amino acid residues within the predicted active site served to change the enzyme from D-cysteine desulfhydrase to 1-aminocyclopropane-1-carboxylate deaminase.
-
-
-
additional information
?
-
Pseudomonas putida, Pseudomonas putida UW4
-
Site-directed mutagenesis shows that altering two amino acid residues at the same positions within the active site of tive site served to change the enzyme from D-cysteine desulfhydrase to deaminase from Pseudomonas putida UW4 the enzyme is converted into D-cysteine desulfhydrase.
-
-
-
additional information
?
-
Escherichia coli W3110
-
L-cysteine is not utilized
-
-
-
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
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
B2MWN0
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
additional product suggested
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
more sulfide than pyruvate and ammonium formed
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
-
more sulfide than pyruvate and ammonium formed
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
plant sulfur metabolism
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
plant sulfur metabolism
additional product suggested, more sulfide than pyruvate and ammonium formed
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
-
physiological function unknown, detoxification of D-cysteine suggested, contributes to utilization of D-cysteine as sulfur source
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
Pseudomonas putida CR 1-1
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
Pseudomonas putida UW4
-
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
Escherichia coli W3110
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
Escherichia coli W3110
-
-
-
?
D-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
Phormidium uncinatum CU 1462/7
-
-
-
?
additional information
?
-
-
extracellular production of hydrogen selenide accounts for thiol-assisted toxicity of selenite against Saccharomyces cerevisiae
-
-
-
additional information
?
-
B2MWN0
Site-directed mutagenesis shows that altering only two amino acid residues within the predicted active site served to change the enzyme from D-cysteine desulfhydrase to 1-aminocyclopropane-1-carboxylate deaminase.
-
-
-
additional information
?
-
Pseudomonas putida, Pseudomonas putida UW4
-
Site-directed mutagenesis shows that altering two amino acid residues at the same positions within the active site of tive site served to change the enzyme from D-cysteine desulfhydrase to deaminase from Pseudomonas putida UW4 the enzyme is converted into D-cysteine desulfhydrase.
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
pyridoxal 5'-phosphate
-
2 mol per mol enzyme
pyridoxal 5'-phosphate
-
-
pyridoxal 5'-phosphate
-
-
pyridoxal 5'-phosphate
-
-
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
(1-aminoethyl)phosphonic acid
-
-
aminooxy acetic acid
-
at 10 microM, IC50 value is 0.0305 mM
aminooxyacetic acid
-
-
aminooxyacetic acid
-
-
aminooxyacetic acid
-
0.005-1 mM, enzymatic activity is progressively inhibited as the aminooxyacetic acid concentration is increased
aminooxyacetic acid
B2MWN0
0.005-1 mM, enzymatic activity is progressively inhibited as the aminooxyacetic acid concentration is increased
D-penicillamine
-
-
hydroxylamine
-
-
hydroxylamine
-
-
hydroxylamine
-
at 10 microM, IC50 value is 0.0159 mM
L-cysteine
-
mixed type inhibition
p-chloromercuribenzoate
-
-
phenylhydrazine
-
-
L-Penicillamine
-
-
additional information
-
no inhibition with EDTA and substrates
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
dithiothreitol
-
probably by preventing autooxidation of substrate
dithiothreitol
-
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.9
-
3-chloro-D-alanine
-
37C, pH 8.0
0.91
-
3-chloro-D-alanine
-
30C, pH 8.0, alpha,beta-elimination
0.29
-
D-allocystathionine
-
30C, pH 8.0
0.05
-
D-Cysteine
-
-
0.14
-
D-Cysteine
-
37C, pH 9.0
0.15
-
D-Cysteine
-
30C, pH 8.0
0.16
-
D-Cysteine
-
37C, pH 9.0
0.21
-
D-Cysteine
B2MWN0
pH 8.0 at 37C
0.25
-
D-Cysteine
-
purified recombinant mature protein
0.3
-
D-Cysteine
-
37C, pH 8.0
0.34
-
D-Cysteine
-
double mutant Pseudomonas putida E295S/L322T
0.27
-
D-Cystine
-
30C, pH 8.0
0.11
-
DL-lanthionine
-
30C, pH 8.0, based on the concentration of the D-isomer, Escherichia coli
0.04
-
DL-selenocystine
-
30C, pH 8.0, based on the concentration of the D-isomer, Escherichia coli
0.28
-
S-benzyl-D-cysteine
-
30C, pH 8.0
0.83
-
S-carboxymethyl-D-cysteine
-
30C, pH 8.0
0.42
-
S-ethyl-D-cysteine
-
30C, pH 8.0
0.5
-
S-methyl-D-cysteine
-
30C, pH 8.0
0.24
-
S-n-butyl-D-cysteine
-
30C, pH 8.0
0.23
-
S-n-propyl-D-cysteine
-
30C, pH 8.0
0.22
-
S-phenyl-D-cysteine
-
30C, pH 8.0
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
281
-
3-chloro-D-alanine
-
37C, pH 8.0
6
-
D-Cysteine
-
-
72
-
D-Cysteine
-
37C, pH 8.0
654
-
D-Cysteine
-
double mutant Pseudomonas putida E295S+L322T
13800
-
D-Cysteine
B2MWN0
-
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0033
-
aminooxyacetic acid
B2MWN0
value bases on non-linear regression
0.53
-
L-cysteine
-
-
IC50 VALUE [mM]
IC50 VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0305
-
aminooxy acetic acid
-
at 10 microM, IC50 value is 0.0305 mM
0.0159
-
hydroxylamine
-
at 10 microM, IC50 value is 0.0159 mM
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.001594
-
-
D-cysteine desulfhydrase activity in WT
0.01
-
-
purified enzyme, 37C, pH 9.0
0.01
-
-
growth of the bacteria with p-nitrophenlysulfate as sulfur source
0.012
-
-
growth of the bacteria with methionine as sulfur source
0.0123
-
-
after partial purification
0.018
-
-
growth of the bacteria with L-cysteine as sulfur; growth of the bacteria with Na2S2O3 and p-nitrophenlysulfate as sulfur; growth of the bacteria with Na2S2O3 as sulfur source
0.02
-
-
growth of the bacteria with MgSO4 and p-nitrophenlysulfate as sulfur source
0.022
-
-
growth of the bacteria with MgSO4 as sulfur source
0.03
-
-
growth of the bacteria with MgSO4 and Na2S2O3 as sulfur source
0.035
-
-
growth of the bacteria with reduced glutathione as sulfur source
0.0356
-
-
D-cysteine desulfhydrase activity in mutant E295S
0.1475
-
-
D-cysteine desulfhydrase activity in double mutant E295S/L322T
0.21
-
-
purified enzyme, 37C, pH 9.0
0.6653
-
B2MWN0
D-cysteine desulfhydrase activity in mutant T386L. Changing the threonine 386 residue alone reduces the activity of the enzyme substantially of about 11.5% of the native recombinant enzyme, although it does not cause complete loss of activity.
5.784
-
B2MWN0
D-cysteine desulfhydrase activity in wild-type
13
-
-
purified enzyme, 30C, pH 8.0, with D-cysteine as substrate
56.3
-
-
purified enzyme, 30C, pH 8.0, with 3-chloro-D-alanine as substrate
additional information
-
B2MWN0
activity below detection, D-cysteine desulfhydrase activity in double mutant S358E/T386L; activity below detection, D-cysteine desulfhydrase activity in mutant S358E. Serine at 358 residue is essential for D-cysteine desulfhydrase activity.
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.6
-
B2MWN0
with estimated 6.7 (pKa1) and 8.9 (pKa2)
8
-
-
assay at
9
-
-
alpha,beta-elimination
9.5
-
-
beta-replacement
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
-
-
assay at
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
loss of activity at 50C, no activity at 60C. The enzyme is also very sensitive to freezing. One freeze-thaw cycle leads to a loss of activity of 75%.
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4.34
-
-
mature protein without N-terminal extension, calculated from amino acid sequence
7.2
-
-
calculated from amino acid sequence
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
B2MWN0
collected at the breaker stage, which is defined as the stage where first spot of pink/red color appears at the blossom end
Manually annotated by BRENDA team
-
primary leaf
Manually annotated by BRENDA team
B2MWN0
collected at the mature green stage
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
Escherichia coli W3110, Pseudomonas putida CR 1-1
-
-
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)
Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)
Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)
Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)
Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)
Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)
Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)
Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)
Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)
Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)
Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
35000
-
-
SDS-PAGE, gel filtration
35150
-
-
calculated from amino acid sequence including N-terminal methionine
36500
-
-
SDS-PAGE
41700
-
-
monomer, mature protein without N-terminal extension, calculated from amino acid sequence
43900
-
-
monomer, calculated from amino acid sequence
51480
-
B2MWN0
recombinant protein is determined by mass spectrometry
67000
-
-
sedimentation equilibrium centrifugation
67000
-
-
gel filtration
70000
-
-
gel filtration
74000
-
-
HPLC gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 42000, SDS-PAGE, unprocessed protein; x * 43000, SDS-PAGE, mature protein; x * 44000, SDS-PAGE, post-translationally modified protein
dimer
-
2 * 36500, SDS-PAGE
dimer
Escherichia coli W3110
-
2 * 36500, SDS-PAGE
-
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
50
-
B2MWN0
The enzyme is found to be very stable with almost 80% of its activity still remaining at 50C
60
-
-
no loss of activity within 30 min incubation, 0.1 M phosphate buffer, pH 8.0
65
-
-
11% loss of activity within 30 min incubation, 0.1 M phosphate buffer, pH 8.0
70
-
-
24% loss of activity within 30 min incubation, 0.1 M phosphate buffer, pH 8.0
75
-
-
40% loss of activity within 30 min incubation, 0.1 M phosphate buffer, pH 8.0
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
glycerol, pyridoxal-5'-phosphate, dithiothreitol and EDTA do not increase the stability of the protein after freezing
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-20C, 10 mM phosphate buffer, pH 7, 0.01 mM pyridoxal phosphate, 0.1 mM DTT, 0.1 mM EDTA 50% glycerol, at least 2 months, no loss of activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
recombinant protein
-
purified from Escherichia coli XL1-Blue cells harboring plasmid pKKyedO
-
partial, does not bind to Blue-Sepharose affinity resin
-
Recombinant protein expressed with an N-terminal 6xHis-tag is purified under native/non-denaturing conditions using Ni-NTA Superflow resin.
-
The recombinant protein expressed with an N-terminal 6xHis-tag is purified under native/non-denaturing conditions using Ni-NTA Superflow resin
B2MWN0
partially by Sephadex G-100 and DEAE column chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression in Escherichia coli
-
overexpressed in host strain, enables Escherichia coli to utilize D-cysteine as sole sulfur source
-
cloning into the pET30a (+) vector at the EcoRV/HindIII sites, all single and double mutants are constructed using a Phusion Site Directed Mutagenesis Kit.
-
Expression of all recombinant proteins and mutants is carried out in Escherichia coli BL21. Altering of only two amino acid residues within the predicted active site served to change the enzyme from D-cysteine desulfhydrase to ACC deaminase.
B2MWN0
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
E295S
-
By site-directed mutagenesis. The Pseudomonas putida UW4 single mutant is constructed using pET30a (+) with the full-length ACC deaminase as the template
E295S/L322T
-
The double mutant is constructed using the E295S mutant as the template.
E295S
Pseudomonas putida UW4
-
By site-directed mutagenesis. The Pseudomonas putida UW4 single mutant is constructed using pET30a (+) with the full-length ACC deaminase as the template
-
E295S/L322T
Pseudomonas putida UW4
-
The double mutant is constructed using the E295S mutant as the template.
-
S358E
B2MWN0
By site-directed mutagenesis. The single mutants of the tomato enzyme are constructed using pET30 Xa/LIC with the full-length putative ACC deaminase as the template
S358E/T386L
B2MWN0
The double mutant is constructed using an additional round of mutagenesis and with the S358E single mutant as the template