Author: ["Paul L. Skatrud","Anthony J. Tietz","Thomas D. Ingolia","Cathleen A. Cantwell","Deborah L. Fisher","Jerry L. Chapman","Stephen W. Queener"]
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Abstract
A recombinant DNA-modified strain of the filamentous fungus Cephalosporium acremonium, strain LU4-79-6, produced more of the antibiotic cephalosporin C than a non-recombinant strain, strain 394-4, from which it was derived by transformation. Strain 394-4, derived from C. acremonium ATCC 11550 by multiple rounds of mutagenesis and screening for improved antibiotic biosynthesis, has been useful for producing cephalosporin C at industrial scale. Strain LU4-79-6 has one insert of pPS56 DNA in its high molecular weight DNA. Plasmid pPS56 includes a dominant hygromycin B resistance marker and a 7 kb BamH1 C. acremonium DNA fragment containing the cefEF gene. The cefEF gene codes for a bifunctional protein that exhibits two sequentially-acting cephalosporin biosynthetic enzyme activities: deacetoxycephalosporin C synthetase (DAOCS) and deacetylcephalosporin C synthetase (DACS). Extracts of strain LU4-79-6 contained ∼2-fold more DAOCS activity than corresponding extracts of its non-recombinant parent. Strain LU4-79-6 excretes less penicillin N, the substrate of DAOCS, than strain 394-4. Resistance to hygromycin B, presence of pPS56 DNA, elevated intracellular DAOCS, decreased penicillin N production, and increased cephalosporin C production are retained after growth in a medium free of hygromycin B. The superiority of the recombinant strain has been confirmed at pilot scale.Cite this article
Skatrud, P., Tietz, A., Ingolia, T. et al. Use of Recombinant DNA to Improve Production of Cephalosporin C By Cephalosporium acremonium. Nat Biotechnol 7, 477–485 (1989). https://doi.org/10.1038/nbt0589-477 