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1.14.20.1: deacetoxycephalosporin-C synthase

This is an abbreviated version!
For detailed information about deacetoxycephalosporin-C synthase, go to the full flat file.

Word Map on EC 1.14.20.1

Reaction

Penicillin N
+
2-oxoglutarate
 +
O2
=
deacetoxycephalosporin C
 +
succinate
 +
CO2
 +
H2O

Synonyms

acDAOC/DACS, cefE, cefEF, Cephalosporin biosynthesis expandase/hydroxylase, DAOC synthase, DAOC/DAC synthase, DAOC/DACS, DAOCS, deacetoxy/deacetylcephalosporin C synthase, deacetoxycephalosporin C synthase, deacetoxycephalosporin-C synthase, deacetoxycephalosporin-C synthetase, deacetoxycephalosporin/deacetylcephalosporin C synthase, expandase, expendase, penicillin N expandase, scDAOCS

ECTree

     1 Oxidoreductases
         1.14 Acting on paired donors, with incorporation or reduction of molecular oxygen
             1.14.20 With 2-oxoglutarate as one donor, and the other dehydrogenated
                1.14.20.1 deacetoxycephalosporin-C synthase

Cloned

Cloned on EC 1.14.20.1 - deacetoxycephalosporin-C synthase

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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
cloning and heterologous expression in Escherichia coli, Streptomyces lividans, Penicillium chrysogenum, Pseudomonas putida and Pichia pastoris, using of commonly used glutathione S-transferase to express DAOCS as a fusion protein
DNA and amino acid sequence determination and analysis, expression as GST-fusion protein in Escherichia coli strain BL21(DE3)
DNA and amino acid sequence determination and analysis, expression of the wild-type enzyme and a C-terminally truncated mutant enzyme lacking 20 amino acids as GST-fusion proteins in Escherichia coli strain BL21(DE3)
expressed in Escherichia coli
expressed in Escherichia coli, Streptomyces lividans, Penicillium chrysogenum, Pseudomonas putida, and Pichia pastoris
expression as glutathione S-transferase fusion protein in Echerichia coli
expression at high levels in Escherichia coli under control of the trc promoter
-
expression in Escherichia coli
expression of wild-type and C-terminally truncated mutant enzymes as N-terminally His-tagged proteins in Escherichia coli strain BL21(DE3)
expression of wild-type and mutant enzymes at high levels in Escherichia coli
-
expression of wild-type and mutant enzymes in Escherichia coli strain BL21
-
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3), subcloning in strain XL-1 Blue
expression of wild-type and mutant enzymes in Escherichia coli strain ESS
-
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)
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
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
gene efeE, cloning in Escherichia coli strain DH5alpha, recombinant expression in Escherichia coli strain K12 (BW25113)
gene efeE, recombinant expression in Escherichia coli strain BL21(DE3)
overexpression as an insoluble and inactive enzyme in granules of recombinant Escherichia coli
-
the attempt to improve the activity of DAOCS by the directed evolution approach is probably the construction of hybrid DAOCS of Streptomyces clavuligerus and Nocardia lactamdurans using in-vivo homologous recombination