Information on EC 2.3.1.28 - chloramphenicol O-acetyltransferase

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

EC NUMBER
COMMENTARY
2.3.1.28
-
RECOMMENDED NAME
GeneOntology No.
chloramphenicol O-acetyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
acetyl-CoA + chloramphenicol = CoA + chloramphenicol 3-acetate
show the reaction diagram
ternary complex mechanism with a rapid equilibrium and essentially random order of addition of substrates
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Acyl group transfer
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
acetyl-CoA:chloramphenicol 3-O-acetyltransferase
-
SYNONYMS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
acetyltransferase, chloramphenicol
-
-
-
-
CAT
-
-
-
-
CAT I
-
-
-
-
CAT III
-
-
-
-
chloramphenicol acetylase
-
-
-
-
chloramphenicol acetyltransferase
-
-
-
-
chloramphenicol transacetylase
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
9040-07-7
-
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
chloramphenicol resistant strains OC, NR, SIN, and T, susceptible control strains DSM8716, ATCC 21536, and ATCC 21537
UniProt
Manually annotated by BRENDA team
infected with either TE/3'2J/CAT or rep5/CAT/26S virus
-
-
Manually annotated by BRENDA team
CATII is encoded by the IncW transmissible plasmid pSA
-
-
Manually annotated by BRENDA team
chloramphenicol-resistant strains which carry the episomal resistance transfer factor R
-
-
Manually annotated by BRENDA team
enzyme form CAT III
-
-
Manually annotated by BRENDA team
enzyme is encoded ny tramsposon Tn2424
-
-
Manually annotated by BRENDA team
JM 109 with plasmid pCM1 containing the enzyme, K12 strains 47, 146, and 570 with chromosome location
-
-
Manually annotated by BRENDA team
multiresistance transposon Tn2424
-
-
Manually annotated by BRENDA team
R-factor-bearing strain W677/HJR 66
-
-
Manually annotated by BRENDA team
strain J53(R387)
-
-
Manually annotated by BRENDA team
strain W677/HJR66
-
-
Manually annotated by BRENDA team
Escherichia coli J53(R387)
strain J53(R387)
-
-
Manually annotated by BRENDA team
Escherichia coli W677/HJR66
strain W677/HJR66
-
-
Manually annotated by BRENDA team
Flavobacterium sp. CB60
CB60
-
-
Manually annotated by BRENDA team
enzymes encoded by plasmid pRI234, pMR375 and pMR385
-
-
Manually annotated by BRENDA team
strain PA 103
Uniprot
Manually annotated by BRENDA team
strain Pahcr, mutations G61S and 105C contribute synergistically to the Pahcr1 resistance phenotype
-
-
Manually annotated by BRENDA team
Pseudomonas aeruginosa PA 103
strain PA 103
Uniprot
Manually annotated by BRENDA team
Pseudomonas aeruginosa Pahcr
strain Pahcr, mutations G61S and 105C contribute synergistically to the Pahcr1 resistance phenotype
-
-
Manually annotated by BRENDA team
encoded by plasmids pSCS6 or pSCS7
-
-
Manually annotated by BRENDA team
enzyme variant C and D
-
-
Manually annotated by BRENDA team
strain C22.1
-
-
Manually annotated by BRENDA team
Staphylococcus aureus C22.1
strain C22.1
-
-
Manually annotated by BRENDA team
enzyme variant B
-
-
Manually annotated by BRENDA team
enzyme type A
-
-
Manually annotated by BRENDA team
type A, type B and type C
-
-
Manually annotated by BRENDA team
enzyme is encoded by plasmid Rms418
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
physiological function
Q6XD71
chloramphenicol resistance
physiological function
-
marker for chloramphenicol resistance
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
-
-
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
in forward reaction formation of a ternary complex by a rapid-equilibrium mechanism, in reverse reaction rapid-equilibrium mechanism with random addition of substrates
-
r
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
acetylates only the biologically active D-threo stereoisomer
+ chloramphenicol 1,3-diacetate
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
inducible enzyme
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
inducible enzyme
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
catabolite repression of CAT synthesis is mediated by a mechanism involving cyclic adenosine 5'-monophosphate
-
-
-
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
inactivates chloramphenicol
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
inactivates chloramphenicol
-
-
-
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
enzymatic inactivation of chloramphenicol
-
-
-
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
all known R factors carrying the CAT gene in enteric bacteria mediate constitutive synthesis of the enzyme
-
-
-
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
Staphylococcus aureus C22.1
-
acetylates only the biologically active D-threo stereoisomer
+ chloramphenicol 1,3-diacetate
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
Staphylococcus aureus C22.1
-
inducible enzyme, inactivates chloramphenicol
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
Pseudomonas aeruginosa Pahcr
-
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
Escherichia coli J53(R387)
-
in forward reaction formation of a ternary complex by a rapid-equilibrium mechanism, in reverse reaction rapid-equilibrium mechanism with random addition of substrates
-
r
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
Escherichia coli W677/HJR66
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
Flavobacterium sp. CB60
-
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 1,3-diacetate
show the reaction diagram
-
-
-
-
?
acetyl-CoA + chloramphenicol 1-acetate
CoA + chloramphenicol 1,3-diacetate
show the reaction diagram
-
no activity
-
-
-
acetyl-CoA + chloramphenicol 1-acetate
CoA + chloramphenicol 1,3-diacetate
show the reaction diagram
-
the enzyme acetylates specifically at the 3-hydroxy position. The diacetylation is possible only because of non-enzymatic interconversion of chloramphenical 3-acetate to chloramphenicol 1-acetate at higher pH values
-
?
acetyl-CoA + chloramphenicol 1-acetate
CoA + chloramphenicol 1,3-diacetate
show the reaction diagram
Flavobacterium sp. CB60
-
no activity
-
-
-
acetyl-CoA + D-threo-1-p-nitrophenyl-2-bromoacetamido-1,3-propanediol
CoA + ?
show the reaction diagram
Staphylococcus aureus, Escherichia coli, Staphylococcus aureus C22.1
-
-
-
-
?
acetyl-CoA + D-threo-1-phenyl-2-dichloroacetamido-1,3-propanediol
CoA + ?
show the reaction diagram
Staphylococcus aureus, Escherichia coli, Staphylococcus aureus C22.1
-
-
-
-
?
chloramphenicol + acetyl-CoA
chloramphenicol 3-acetate + CoA
show the reaction diagram
Q6XD71
-
-
-
?
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
inducible enzyme
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
inducible enzyme
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
catabolite repression of CAT synthesis is mediated by a mechanism involving cyclic adenosine 5'-monophosphate
-
-
-
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
inactivates chloramphenicol
-
-
?
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
inactivates chloramphenicol
-
-
-
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
enzymatic inactivation of chloramphenicol
-
-
-
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
-
all known R factors carrying the CAT gene in enteric bacteria mediate constitutive synthesis of the enzyme
-
-
-
acetyl-CoA + chloramphenicol
CoA + chloramphenicol 3-acetate
show the reaction diagram
Staphylococcus aureus C22.1
-
inducible enzyme, inactivates chloramphenicol
-
-
?
chloramphenicol + acetyl-CoA
chloramphenicol 3-acetate + CoA
show the reaction diagram
Q6XD71
-
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
HgCl2
-
at 0.1 M and 0.3 mM, no influence on the activity of the pSCS6-encoded enzyme. The activity of the pSCS7-encoded enzyme decreases to 10-15%. At 1.0 mM HgCl2, the pSCS6-encoded enzyme retains 50% of ist original activity, complete inactivation of the enzyme from pSCS7-encoded enzyme
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1,5-Difluoro-2,4-dinitrobenzene
-
-
4,4'-dithiodipyridine
-
-
5,5'-dithiobis(2-nitrobenzoic acid)
-
-
5,5'-dithiobis(2-nitrobenzoic acid)
-
0.025 mM, 82% inhibition
5,5'-dithiobis(2-nitrobenzoic acid)
-
-
5,5'-dithiobis(2-nitrobenzoic acid)
-
-
Crystal violet
-
strong, competitive for chloramphenicol, non-competitive for acetyl-CoA
EDTA
-
0.1 mM, 24% inhibition
ethyl violet
-
strong, competitive for chloramphenicol, non-competitive for acetyl-CoA
Hg2+
-
0.01 mM, complete inhibition
I2
-
0.1 mM, 20% inhibition
methyl methanethiolsulfonate
-
-
PCMB
-
0.1 mM, complete inhibition
Zn2+
-
0.1 mM, 15% inhibition
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
acetyl-CoA
Q6XD71
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.028
acetyl-CoA
-
wild-type enzyme
0.0465
acetyl-CoA
-
enzyme variant D
0.055
acetyl-CoA
-
enzyme variant CAT I
0.056
acetyl-CoA
-
enzyme from plasmid pSCS7
0.0561
acetyl-CoA
-
-
0.0573
acetyl-CoA
-
-
0.059
acetyl-CoA
-
enzyme from plasmid pSCS6
0.0609
acetyl-CoA
-
enzyme variant C
0.065
acetyl-CoA
-
-
0.065
acetyl-CoA
-
-
0.083
acetyl-CoA
-
-
0.093
acetyl-CoA
-
enzyme variant CATIII
0.094
acetyl-CoA
-
enzyme variant CATA1
0.156
acetyl-CoA
-
-
0.161
acetyl-CoA
-
enzyme variant CATB1
0.164
acetyl-CoA
-
enzyme variant CATB5
0.165
acetyl-CoA
-
enzyme variant CATB3
0.797
acetyl-CoA
-
mutant enzyme G61S/Y105C
0.806
acetyl-CoA
-
mutant enzyme G61S
0.809
acetyl-CoA
-
mutant enzyme G61S/Y105C
0.82
acetyl-CoA
-
mutant enzyme Y105C
0.821
acetyl-CoA
-
enzyme variant CATB7
0.00247
Chloramphenicol
-
enzyme variant C
0.0025
Chloramphenicol
-
enzyme from plasmid pSCS6
0.00258
Chloramphenicol
-
-
0.0027
Chloramphenicol
-
-
0.0027
Chloramphenicol
-
enzyme from plasmid pSCS7
0.0027
Chloramphenicol
-
enzyme variant D
0.00272
Chloramphenicol
-
-
0.0061
Chloramphenicol
-
-
0.0093
Chloramphenicol
-
-
0.01
Chloramphenicol
-
-
0.0103
Chloramphenicol
-
-
0.011
Chloramphenicol
-
enzyme variant CATA1
0.011
Chloramphenicol
-
enzyme variant CAT I
0.012
Chloramphenicol
-
enzyme variant CAT III
0.0166
Chloramphenicol
-
-
0.0175
Chloramphenicol
-
-
0.0205
Chloramphenicol
-
-
0.0215
Chloramphenicol
-
-
0.025
Chloramphenicol
-
wild-type enzyme
0.03
Chloramphenicol
-
-
0.031
Chloramphenicol
-
-
0.033
Chloramphenicol
-
-
0.0345
Chloramphenicol
-
-
0.136
Chloramphenicol
-
enzyme variant CATB5
0.14
Chloramphenicol
-
enzyme variant CATB1
0.156
Chloramphenicol
-
enzyme variant CATB3
0.795
Chloramphenicol
-
mutant enzyme G61S/Y105C
0.798
Chloramphenicol
-
wild-type enzyme
0.8 - 11
Chloramphenicol
-
mutant enzyme G61S
0.809
Chloramphenicol
-
mutant enzyme Y105C
0.812
Chloramphenicol
-
enzyme variant CATB7
5
Chloramphenicol
-
-
0.0037
D-threo-1-p-nitrophenyl-2-bromoacetamido-1,3-propanediol
-
-
0.022
D-threo-1-p-nitrophenyl-2-bromoacetamido-1,3-propanediol
-
-
0.021
D-threo-1-phenyl-2-dichloroacetamido-1,3-propanediol
-
-
0.068
D-threo-1-phenyl-2-dichloroacetamido-1,3-propanediol
-
-
additional information
additional information
-
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
-
-
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
-
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
-
additional information
-
-
additional information
-
-
additional information
-
relative activity: 94.9% (wild-type), 34.6% (fluorinated CAT), 89% (mutant L158I), 51.7% (fluorinated mutant L158I)
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5 - 9
-
about 50% of maximal activity at pH 5.0 and 9.0
6 - 8.8
-
pH 6.0: about 55% of maximal activity, pH 8.8: about 85% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
enzyme expression is increased at 42C. At 42C the enzyme is still soluble and active
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30 - 45
-
30C: about 20% of maximal activity, 45C: about 55% of maximal activity
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
-
Culex pipiens infected with either TE/3'2J/CAT or rep5/CAT/26S virus
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
26660
Q6XD71
predicted from sequence analysis, 347 bp, 228 amino acid protein, chromosomal location
703753
69000
-
gel filtration
486718
70000
-
gel filtration
486715
78000
-
sucrose density gradient centrifugation
486713
78000
-
gel filtration
486734, 658535
80000
-
gel filtration
486722
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
-
x * 23351, electrospray injection mass spectrometry
?
-
x * 24150, enzyme encoded by plasmid pRI234, x * 24400, enzyme encoded by plasmid pMR375 and plasmid pMR385
tetramer
-
1 * 19500 + 1 * 19000 + 1 * 18000 + 1 * 17500, SDS-PAGE
trimer
-
3 * 22000, SDS-PAGE
trimer
-
3 * 23000, SDS-PAGE
trimer
-
3 * 26600, SDS-PAGE
trimer
Flavobacterium sp. CB60
-
3 * 26600, SDS-PAGE
-
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structure of CATI in the unbound (apo) and CAM-bound forms are refined to 3.2 A and 2.9 A resolution, respectively
P62577
hanging drop vapour diffusion method
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
53
-
0.1 mM chloramphenicol and 0.1 mM acetyl-CoA, 50 mM Tris-HCl buffer, pH 7.8, half-life: 100 min
486718
55
-
,30 min, 82% loss of activity, wild-type enzyme
486728
60
-
nearly complete inactivation within 10 min
486722
70
-
15 min, 40% loss of activity of enzyme variant from plasmid pSCS6, 45% loss of activity of enzyme variant pSCS7
486715
75
-
rapid inactivation
486713
75
-
remarkably resistant
486713
75
-
stable
486714
75
-
-
486714
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
loss of activity by freezing and thawing can be prevented by adding 10% glycerol
-
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
photooxidation of enzyme type C in presence of methylene blue, progressive loss of activity with increase in pH, inclusion of 1 mM chloramphenicol affords complete protection against loss of activity
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486724
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20 C, activity in elution buffer is stable for at least 1 year
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-20 C, stable for several months
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-80 C, 20% glycerol, stable for more than 3 months
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ELP (elastin-like polypeptide) fusion tag technique to purify ultra-low levels of chloramphenicol acetyltransferase-ELP fusion protein
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enzyme variant C and D
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enzyme variant B
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enzyme variant A
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plasmid Rms418 is transferred from Vibrio anguillarum to E. coli K-12
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
PCR-amplification, electrotransformation into Escherichia coli DH10B to test for chloramphenicol inactivation with Micrococcus lutes ATCC 9341 as indicator organism
Q6XD71
cloning of mutant cat-86 in pTB361 and transformation of Escherichia coli JM109
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expressed in Escherichia coli
P62577
cloning of enzyme form CATB7
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overexpression in Escherichia coli harboring the plasmid pYT1
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PCR-amplification, clinging into Escherichia coli MC1061, electrotransformaion of Leuconostoc mesenteroides DRC (isolated from fermenting salted Chinese cabbage/Kimchi)
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plasmid Rms418 is transferred from Vibrio anguillarum to Escherichia coli K-12
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
A203G
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mutant enzyme is less stable than wild-type enzyme
A203I
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mutant enzyme is more thermostable than wild-type
I191V
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mutant enzyme is less stable than wild-type enzyme
C214A
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95% of the in vivo produced mutant polypeptide is soluble compared to 90% for the wild-type enzyme. Mutant enzyme loses 31% of activity after 30 min at 65C compared to 15% for the wild-type enzyme
C214D
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50% of the in vivo produced mutant polypeptide is soluble compared to 90% for the wild-type enzyme. Mutant enzyme loses 85% of activity after 30 min at 65C compared to 15% for the wild-type enzyme
C214E
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75% of the in vivo produced mutant polypeptide is soluble compared to 90% for the wild-type enzyme. Mutant enzyme loses 84% of activity after 30 min at 65C compared to 15% for the wild-type enzyme
C214F/G219S
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95% of the in vivo produced mutant polypeptide is soluble compared to 90% for the wild-type enzyme. Mutant enzyme loses 81% of activity after 30 min at 65C compared to 15% for the wild-type enzyme
C214G
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80% of the in vivo produced mutant polypeptide is soluble compared to 90% for the wild-type enzyme. Mutant enzyme loses 44% of activity after 30 min at 65C compared to 15% for the wild-type enzyme
C214L
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100% of the in vivo produced mutant polypeptide is soluble compared to 90% for the wild-type enzyme. Mutant enzyme loses 33% of activity after 30 min at 65C compared to 15% for the wild-type enzyme
C214P
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95% of the in vivo produced mutant polypeptide is soluble compared to 90% for the wild-type enzyme. Mutant enzyme loses 88% of activity after 30 min at 65C compared to 15% for the wild-type enzyme
C214Q
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95% of the in vivo produced mutant polypeptide is soluble compared to 90% for the wild-type enzyme. Mutant enzyme loses 73% of activity after 30 min at 65C compared to 15% for the wild-type enzyme
C214R
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55% of the in vivo produced mutant polypeptide is soluble compared to 90% for the wild-type enzyme. Mutant enzyme loses 84% of activity after 30 min at 65C compared to 15% for the wild-type enzyme
C214S
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95% of the in vivo produced mutant polypeptide is soluble compared to 90% for the wild-type enzyme. Mutant enzyme loses 32% of activity after 30 min at 65C compared to 15% for the wild-type enzyme
C214T
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90% of the in vivo produced mutant polypeptide is soluble compared to 90% for the wild-type enzyme. Mutant enzyme loses 59% of activity after 30 min at 65C compared to 15% for the wild-type enzyme
C214V
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95% of the in vivo produced mutant polypeptide is soluble compared to 90% for the wild-type enzyme. Mutant enzyme loses 45% of activity after 30 min at 65C compared to 15% for the wild-type enzyme
C214W
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50% of the in vivo produced mutant polypeptide is soluble compared to 90% for the wild-type enzyme. Mutant enzyme loses 70% of activity after 30 min at 65 C compared to 15% for the wild-type enzyme
C214Y
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90% of the in vivo produced mutant polypeptide is soluble compared to 90% for the wild-type enzyme. Mutant enzyme loses 81% of activity after 30 min at 65C compared to 15% for the wild-type enzyme
CATIII (F24A/Y25F/L29A)
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Km-value for acetyl-CoA is 0.095 mM compared to 0.093 mM for wild-type CATIII, Km-value for chloramphenicol is 0.023 mM compared to 0.012 mM for the wild-type CATIII, turnover number is 30% of the wild-type enzyme CAT III
CATIII(K14E/H195A/K217A)
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no activity
CATIII(Q92C/N146F/Y169F/I172V)
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Km-value for acetyl-CoA is 0.165 mM compared to 0.093 mM for wild-type CATIII, Km-value for chloramphenicol is 0.02 mM compared to 0.012 mM for the wild-type CATIII, turnover number is 60% of the wild-type enzyme CAT III
K14/K217E
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Km-value for acetyl-CoA is 0.166 mM compared to 0.093 mM for wild-type CATIII, Km-value for chloramphenicol is 0.017 mM compared to 0.012 mM for the wild-type CATIII, turnover number is 87% of the wild-type enzyme CAT III
L145F
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folding of chloramphenicol acetyltransferase is hampered by deletion of the carboxy-terminal tail including the last residue of the carboxy-terminal alpha-helix. Such truncated CAT polypeptides quantitatively aggregate into cytoplasmic inclusion bodies, which results in absence of chloramphenicol-resistant phenotype for the producing host. Introduction of Phe at amino acid position 145 improves the ability of the protein to fold into a soluble, enzymatically active conformation
L158I
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fluorinated mutant expressed in trifluoroleucine shows enhanced thermostability compared to CAT T (CAT expressed in trifluoroleucine), suggesting that trifluoroleucine at position 158 contributes to a portion of the observed loss in thermostability upon global fluorination. Relative activity: 89% (non-fluorinated mutant), 51.7% (fluorinated mutant)
L208I
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fluorinated mutant expressed in trifluoroleucine shows loss in thermostability
[CATI (H195A)]2[CATIII(K14E/K217E)]
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hybrid trimer, Km-value for acetyl-CoA is 0.072 mM compared to 0.093 mM for wild-type CATIII, Km-value for chloramphenicol is 0.018 mM compared to 0.012 mM for the wild-type CATIII, turnover number is 14% of the wild-type enzyme CAT III
[CATIII]2[CATIII(K14E/H195A/K217A)]
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Km-value for acetyl-CoA is 0.143 mM compared to 0.093 mM for wild-type CATIII, Km-value for chloramphenicol is 0.016 mM compared to 0.012 mM for the wild-type CATIII, turnover number is 80% of the wild-type enzyme CAT III
[CATIII][CATIII(K14E/H195A/K217A)]2
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Km-value for acetyl-CoA is 0.198 mM compared to 0.093 mM for wild-type CATIII, Km-value for chloramphenicol is 0.02 mM compared to 0.012 mM for the wild-type CATIII, turnover number is 82% of the wild-type enzyme CAT III
[CATI][CATIII(K14E/H195A/K217E)]2
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hybrid trimer, Km-value for acetyl-CoA is 0.107 mM compared to 0.093 mM for wild-type CATIII, Km-value for chloramphenicol is 0.02 mM compared to 0.012 mM for the wild-type CATIII, turnover number is 50% of the wild-type enzyme CAT III
G61S
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G61S and Y105C contrtibute synergistically to the resistance phenotype of strain PAhcr1
G61S/Y105C
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G61S and Y105C contrtibute synergistically to the resistance phenotype of strain PAhcr1
Y105C
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G61S and Y105C contrtibute synergistically to the resistance phenotype of strain PAhcr1
G61S
Pseudomonas aeruginosa Pahcr
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G61S and Y105C contrtibute synergistically to the resistance phenotype of strain PAhcr1
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G61S/Y105C
Pseudomonas aeruginosa Pahcr
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G61S and Y105C contrtibute synergistically to the resistance phenotype of strain PAhcr1
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Y105C
Pseudomonas aeruginosa Pahcr
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G61S and Y105C contrtibute synergistically to the resistance phenotype of strain PAhcr1
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Y33F/A203V
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mutant enzyme is more thermostable than wild-type
additional information
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Bacillus subtilis cells expressing a hybrid protein (LvsSS-Cat) consisting of the Bacillus amyloliquefaciens levansucrose signal peptide fused to Bacillus pumilus chloramphenicol acetyltransferase are unable to export cat protein into the growth medium. A series of tripartite protein fusion is constructed by inserting various length of the cat sequences between the levansucrase signal peptide and staphylococcal protein A or Escherichia coli alkaline phosphatase. Biochemical characterization of the various Cat protein fusion reveales that multiple regions in the cat protein are causing the export defect
L821I
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fluorinated mutant expressed in trifluoroleucine shows loss in thermostability
additional information
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in soluble CATI(1-211)(X3) mutants nearly all amino acid residues are tolerated at position 212 and 213. This reflects the relative lack of impotance of these residues in the folding and/or stabilization of CAT. Substitutions at position 214 do not dramatically alter the biological activity of wild-type CATI
additional information
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replacement of all the leucine residues in the enzyme chloramphenicol acetyltransferase with the analog, 5',5',5'-trifluoroleucine, results in the maintenance of enzymatic activity under ambient temperatures as well as an enhancement in secondary structure but loss in stability against heat and denaturants or organic co-solvents
additional information
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residue-specific incorporation of T into chloramphenicol acetyltransferase (CAT) results in a loss of thermostability. Relative activity: 34.6% (fluorinated CAT)
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
medicine
Q6XD71
probiotic mixture for the prevention of gastrointestinal side effects due to oral antibiotic therapy, no resistance transfer to Enterococcus faecalis JH2-2, Enterococcus faecium HM1070 (both resistant to rifampin and fusidic acid) and Bacillus subtilis UCN19 (resistant to ciprofloxacin) in mating experiments
analysis
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valuable tool in studies of eukaryotic gene expression, quantitative aspects of the use of bacterial chloramphenicol acetyltransferase as a reporter system in the yeast Saccharomyces cerevisiae
molecular biology
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the chloramphenicol acetyl transferase gene serves as integration target for the eukaryotic mariner transposon Mos1 regardless of the location (chromosome or plasmid), as tested with in vitro and bacterial transposition assays
molecular biology
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single-chain variable fragment (scFv) phages are selected with affinity for CAT. Surface plasmon resonance analyses shows that the tested scFv phages have an affinity for CAT with a dissociation constant (Kd) around 1 microM. The selected scFv phages can be used as capture elements in a highly sensitive sandwich ELISA to detect CAT concentration as low as 0.1 ng/ml or 4 pM
biotechnology
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the development of the chloramphenicol acetyltransferase gene cat as a new selectable marker for plastid transformation is reported. By selecting for chloramphenicol resistance, tobacco chloroplast transformants are readily obtained. Transplastomic lines quickly reach the homoplasmic state, accumulate the chloramphenicol acetyltransferase enzyme to high levels and transmit their plastid transgenes maternally into the next generation
biotechnology
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monitoring of starter (Leuconostoc mesenteroides DRC) growth in lactic acid-fermented Kimchi (salted Chinese cabbage) for predicting starter predominance during fermentation by stable transformation of the chloramphenicol acetyltransferase gene into the chromosomal DNA with a transposon vector