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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
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
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
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