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Literature summary for 1.14.20.1 extracted from

  • Fan, K.; Lin, B.; Tao, Y.; Yang, K.
    Engineering deacetoxycephalosporin C synthase as a catalyst for the bioconversion of penicillins (2017), J. Ind. Microbiol. Biotechnol., 44, 705-710 .
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
ascorbate required Streptomyces clavuligerus
ascorbate required Acremonium chrysogenum
ascorbate required Amycolatopsis lactamdurans

Application

Application Comment Organism
synthesis bioconversion of penicillins to cephalosporins using deacetoxycephalosporin C synthase (DAOCS) is an alternative and environmentally friendly process for production of 7-aminodeacetoxycephalosporanic acid (7-ADCA), a key intermediate of many clinically useful semisynthetic cephalosporins Streptomyces clavuligerus
synthesis bioconversion of penicillins to cephalosporins using deacetoxycephalosporin C synthase (DAOCS) is an alternative and environmentally friendly process for production of 7-aminodeacetoxycephalosporanic acid (7-ADCA), a key intermediate of many clinically useful semisynthetic cephalosporins Acremonium chrysogenum
synthesis bioconversion of penicillins to cephalosporins using deacetoxycephalosporin C synthase (DAOCS) is an alternative and environmentally friendly process for production of 7-aminodeacetoxycephalosporanic acid (7-ADCA), a key intermediate of many clinically useful semisynthetic cephalosporins Amycolatopsis lactamdurans

Cloned(Commentary)

Cloned (Comment) Organism
gene cefE, recombinant expression in Escherichia coli BW25311. Recombinant expression of a hybrid enzyme,with parts from the enzymes of Streptomyces clavuligerus and Nocardia lactamdurans, in Streptomyces lividans strain W25, the recombinant enzyme is capable of converting penicillin G to G-7-aminodeacetoxycephalosporanic acid (G-7-ADCA) Amycolatopsis lactamdurans
gene cefE, recombinant expression in Escherichia coli strain BW25311, functional recombinant expression of scDAOCS in Penicillium chrysogenum leading to expandase activity for penicillin N in the transfected cells. But only penicillin V is produced under the fermentation conditions, indicating that the industrial application of DAOCS is restricted by its low activities toward penicillin analogues other than penicillin N. In Penicillium chrysogenum strains expressing scDAOCS, a fermentation process is developed to produce adipyl-7-ADCA and adipyl-7-ACA, which can be converted to 7-ADCA and 7-ACA by glutaryl acylase. Recombinant expression of a hybrid enzyme,with parts from the enzymes of Streptomyces clavuligerus and Nocardia lactamdurans, in Streptomyces lividans strain W25, the recombinant enzyme is capable of converting penicillin G to G-7-aminodeacetoxycephalosporanic acid (G-7-ADCA). Recombinant expression of Streptomyces clavuligerus scDAOCS in a cefEF-disrupted cephalosporin C production Acremonium chrysogenum strain, which converts penicillin N accumulated in the disruption strain to deacetoxycephalosporin C Streptomyces clavuligerus
gene cefEF, recombinant expression in Escherichia coli, functional recombinant expression of acDAOCS in Penicillium chrysogenum strains leads to a fermentation process developed to produce adipyl-7-ADCA and adipyl-7-ACA, which can be converted to 7-ADCA and 7-ACA by glutaryl acylase Acremonium chrysogenum

Protein Variants

Protein Variants Comment Organism
C155Y/Y184H/S261M/V275I/C281Y/I305M site-directed mutagenesis, the mutant shows 87fold increased kcat/Km for penicillin G compared to the wild-type Streptomyces clavuligerus
C155Y/Y184H/V275I/C281Y site-directed mutagenesis, quaternary mutant from error-prone PCR, the mutant shows 41fold increased kcat/Km for penicillin G compared to the wild-type Streptomyces clavuligerus
M73T/C155Y/Y184H/T213V/V275I/C281Y/I305M site-directed mutagenesis, the mutant shows 22fold increased specific activity and 81fold increased kcat/Km for penicillin G compared to the wild-type Streptomyces clavuligerus
additional information rational mutational approach to generate an engineered strain for enhanced production of from penicillin G, e.g. mutant FF8 from gene shuffling with 118fold increase in kcat/Km Streptomyces clavuligerus
additional information recombinant expression of Streptomyces clavuligerus scDAOCS in a cefEF-disrupted cephalosporin C production Acremonium chrysogenum strain, which converts penicillin N accumulated in the disruption strain to deacetoxycephalosporin C Acremonium chrysogenum

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ dependent on, required for catalysis Streptomyces clavuligerus
Fe2+ dependent on, required for catalysis Acremonium chrysogenum
Fe2+ dependent on, required for catalysis Amycolatopsis lactamdurans

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
penicillin N + 2-oxoglutarate + O2 Streptomyces clavuligerus
-
deacetoxycephalosporin C + succinate + CO2 + H2O
-
?
penicillin N + 2-oxoglutarate + O2 Acremonium chrysogenum
-
deacetoxycephalosporin C + succinate + CO2 + H2O
-
?
penicillin N + 2-oxoglutarate + O2 Amycolatopsis lactamdurans
-
deacetoxycephalosporin C + succinate + CO2 + H2O
-
?
penicillin N + 2-oxoglutarate + O2 Streptomyces clavuligerus ATCC 27064
-
deacetoxycephalosporin C + succinate + CO2 + H2O
-
?

Organism

Organism UniProt Comment Textmining
Acremonium chrysogenum P11935 i.e. Cephalosporium acremonium
-
Amycolatopsis lactamdurans Q03047
-
-
Streptomyces clavuligerus P18548
-
-
Streptomyces clavuligerus ATCC 27064 P18548
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme from Escherichia coli Streptomyces clavuligerus
recombinant enzyme from Escherichia coli Acremonium chrysogenum
recombinant enzyme from Escherichia coli Amycolatopsis lactamdurans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information purified recombinant enzyme DAOCS expressed in Escherichia coli catalyzes the ring expansion of penicillin N but shows no hydroxylation activity for DAOC, EC 1.14.11.26 Streptomyces clavuligerus ?
-
?
additional information purified recombinant enzyme DAOCS expressed in Escherichia coli catalyzes the ring expansion of penicillin N but shows no hydroxylation activity for DAOC, EC 1.14.11.26 Streptomyces clavuligerus ATCC 27064 ?
-
?
penicillin G + 2-oxoglutarate + O2
-
Streptomyces clavuligerus 7-phenylacetylaminocephalosporanate + succinate + CO2 + H2O
-
?
penicillin G + 2-oxoglutarate + O2
-
Acremonium chrysogenum 7-phenylacetylaminocephalosporanate + succinate + CO2 + H2O
-
?
penicillin G + 2-oxoglutarate + O2
-
Amycolatopsis lactamdurans 7-phenylacetylaminocephalosporanate + succinate + CO2 + H2O
-
?
penicillin G + 2-oxoglutarate + O2
-
Streptomyces clavuligerus ATCC 27064 7-phenylacetylaminocephalosporanate + succinate + CO2 + H2O
-
?
penicillin N + 2-oxoglutarate + O2
-
Streptomyces clavuligerus deacetoxycephalosporin C + succinate + CO2 + H2O
-
?
penicillin N + 2-oxoglutarate + O2
-
Acremonium chrysogenum deacetoxycephalosporin C + succinate + CO2 + H2O
-
?
penicillin N + 2-oxoglutarate + O2
-
Amycolatopsis lactamdurans deacetoxycephalosporin C + succinate + CO2 + H2O
-
?
penicillin N + 2-oxoglutarate + O2 ring expansion of penicillin N Streptomyces clavuligerus deacetoxycephalosporin C + succinate + CO2 + H2O
-
?
penicillin N + 2-oxoglutarate + O2
-
Streptomyces clavuligerus ATCC 27064 deacetoxycephalosporin C + succinate + CO2 + H2O
-
?
penicillin N + 2-oxoglutarate + O2 ring expansion of penicillin N Streptomyces clavuligerus ATCC 27064 deacetoxycephalosporin C + succinate + CO2 + H2O
-
?

Synonyms

Synonyms Comment Organism
cefE
-
Streptomyces clavuligerus
cefE
-
Amycolatopsis lactamdurans
cefEF
-
Acremonium chrysogenum
DAOCS
-
Streptomyces clavuligerus
DAOCS
-
Acremonium chrysogenum
DAOCS
-
Amycolatopsis lactamdurans
deacetoxycephalosporin C synthase
-
Streptomyces clavuligerus
deacetoxycephalosporin C synthase
-
Acremonium chrysogenum
deacetoxycephalosporin C synthase
-
Amycolatopsis lactamdurans
expandase
-
Acremonium chrysogenum

General Information

General Information Comment Organism
evolution the enzyme belongs to the iron- and 2-oxoglutarate-dependent oxidases Streptomyces clavuligerus
evolution the enzyme belongs to the iron- and 2-oxoglutarate-dependent oxidases Acremonium chrysogenum
evolution the enzyme belongs to the iron- and 2-oxoglutarate-dependent oxidases Amycolatopsis lactamdurans
physiological function Cephalosporium acremonium expandase is a bifunctional enzyme, penicillin expandase, which catalyzes the ring expansion of penicillin N and the hydroxylation of deacetoxycephalosporin C (DAOC) to deacetylcephalosporin C (DAC). 2-oxoglutarate, Fe2+ and O2 are required for its catalytic activity. The enzyme is encoded by gene cefEF Acremonium chrysogenum
physiological function Cephalosporium acremonium expandase is a bifunctional enzyme, which catalyzes the ring expansion of penicillin N and the hydroxylation of deacetoxycephalosporin C (DAOC) to deacetylcephalosporin C (DAC). 2-oxoglutarate, Fe2+ and O2 are required for its catalytic activity. The enzyme is encoded by gene cefEF Amycolatopsis lactamdurans