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Use of Recombinant DNA to Improve Production of Cephalosporin C By Cephalosporium acremonium

Bio/Technology volume 7, pages 477–485 (1989)Cite this article

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.

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Author notes

  1. Stephen W. Queener: Corresponding Author

Authors and Affiliations

  1. Lilly Research Laboratories, a Division of Eli Lilly and Company, Indianapolis, IN, 46285

    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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  3. Thomas D. Ingolia
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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

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