Information on EC 2.3.1.175 - deacetylcephalosporin-C acetyltransferase

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The expected taxonomic range for this enzyme is: Acremonium chrysogenum

EC NUMBER
COMMENTARY hide
2.3.1.175
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RECOMMENDED NAME
GeneOntology No.
deacetylcephalosporin-C acetyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
acetyl-CoA + deacetylcephalosporin C = CoA + cephalosporin C
show the reaction diagram
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-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Biosynthesis of antibiotics
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cephalosporin C biosynthesis
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Metabolic pathways
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Penicillin and cephalosporin biosynthesis
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SYSTEMATIC NAME
IUBMB Comments
acetyl-CoA:deacetylcephalosporin-C O-acetyltransferase
This enzyme catalyses the final step in the biosynthesis of cephalosporin C.
CAS REGISTRY NUMBER
COMMENTARY hide
57827-76-6
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
acetyl-CoA + deacetylcephalosporin C
CoA + cephalosporin C
show the reaction diagram
additional information
?
-
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expression is regulated by methionine
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-
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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
acetyl-CoA + deacetylcephalosporin C
CoA + cephalosporin C
show the reaction diagram
additional information
?
-
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expression is regulated by methionine
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-
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,6-dihydroxybenzoic acid
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0.1 mM, 27% inhibition
coenzyme A
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0.1 mM, 61% inhibition
deacetoxycephalosporin C
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0.1 mM, 57% inhibition
Penicillin N
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0.1 mM, 14% inhibition
pyruvate
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0.1 mM, 37% inhibition
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.1 - 1.05
acetyl-CoA
0.04 - 0.29
deacetylcephalosporin C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.7
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deacetylcephalosporin-C acetyltransferase activity in the host strain Penicillium chrysogenum TA64
7.2
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deacetylcephalosporin-C acetyltransferase activity in the host strain Penicillium chrysogenum TA71
25.8
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deacetylcephalosporin-C acetyltransferase activity in the host strain Penicillium chrysogenum TA2
30
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deacetylcephalosporin-C acetyltransferase activity in the host strain Penicillium chrysogenum TA98
additional information
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0.0000378 mg cephalosporin C per min per mg
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 8
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no significant influence of pH-value on enzyme activity is measured after 20 min
7
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activity assay
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 8.5
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pH 6.0: about 60% of maximal activity, pH 8.5: about 50% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
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activity assay
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
15 - 40
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15°C: about 45% of maximal activity, 40°C: about 75% of maximal activity
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
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chromatofocusing
4.3
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isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
49000
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1 * 49000, SDS-PAGE
50000
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gel filtration
55000
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gel filtration
70000
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gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
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SDS-PAGE resolves two protein bands with molecular weights of 27000 Da and 14000 Da
monomer
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1 * 49000, SDS-PAGE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
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DCPC-ATF is synthesized as a single-chain precursor protein and is then converted to the two constituent subunit DCPC-ATF by proteolytic processing
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
different structures are solved, SeMet-DAC-AT with acetyl-CoA to a resolution of 2.4 A, DAC-AT with acetyl-CoA to 2.2 A, DAC-AT with DAC/cephalosporin C to 2.5 A, and DAC-AT with DAC to 2.6 A resolution
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
using a gravity-flow column containing chitin beads, cleavage of the chitin-binding domain and intein fusion partner is performed
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
constructions starting at each of the three in-frame ATG codons of the Acremonium chrysogenum cefG gene (Met1, Met46 and Met60) are expressed in Escherichia coli, obtaining proteins of 49000 Da, 44000 Da and 43000 Da, respectively. All three proteins show deacetylcephalosporin C acetyltransferase activity, separately or in combination
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expression in Penicillium chrysogenum
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expression in yeast, the leader peptide is not essential for expression of the enzyme activity
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expression of the cefG gene from the promoters of 1. the glyceraldehyde-3-phosphate dehydrogenase gene of Aspergillus nidulans, 2. the glucoamylase gene of Aspergillus niger, 3. the glutamate dehydrogenase and 4. the isopenicillin N synthase genes of Penicillium chrysogenum, leads to very high steady-state levels of cefG transcript and to increased deacetylcephalosporin-C acetyltransferase protein concentration
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into the pTWIN vector for expression in Escherichia coli BL21DE3 cells
Penicillium chrysogenum strains expressing cefD1, cefD2, cefEF, and cefG genes cloned form Acremonium chrysogenum are constructed, deacetylcephalosporin-C acetyltransferase is coded by the cefG gene
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
biotechnology
synthesis
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production of acetylcephalosporin C by gene expression in Penicillium chrysogenum. Recombinant strains secrete significant amounts of deacetylcephalosporin C, but acetylcephalosporin C is not detected in culture broth. Even when accumulating intracellularly, acetylcephalosporin C is not found extracellularly