Cloning and Expression of the Fungal Expandase/hydroxylase Gene Involved in Cephalosporin Biosynthes
Author: ["Suellen M. Samson","Joe E. Dotzlaf","Mark L. Slisz","Gerald W. Becker","Richard M. Van Frank","Loraine E. Veal","Wu-Kuang Yeh","James R. Miller","Stephen W. Queener","Thomas D. Ingolia"]
Publication: Bio/Technology
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Abstract
Cephalosporins are antibiotics characterized by a cephem ring system in which a beta–lactam ring is fused to a dihydrothiazine ring. The cephem ring system is synthesized by expansion of the five–membered thiazolidine ring of the penicillin to the six–membered dihydrothiazine ring. The enzyme that catalyzes this reaction, deacetoxycephalosporin C synthetase (DAOCS)—often called expandase—has been purified from C. acremonium. During purification DAOCS activity never separated from and remained in constant ratio to deacetylcephalosporin C synthetase (DACS, or hydroxylase) activity. A partial amino acid sequence of the C. acremonium DAOCS/DACS protein has been obtained and this information used to clone a corresponding gene. The open reading frame observed upon sequencing this gene was expressed in E. coli and both DAOCS and DACS activities were found in extracts of these E. coli cells, providing unambiguous proof of the bifunctional nature of the DAOCS/DACS polypeptide from C. acremonium. The cloned DAOCS/DACS gene represents an important tool for enhancing production of known cephalosporins and for synthesis of novel cephalosporins.
Cite this article
Samson, S., Dotzlaf, J., Slisz, M. et al. Cloning and Expression of the Fungal Expandase/hydroxylase Gene Involved in Cephalosporin Biosynthesis. Nat Biotechnol 5, 1207–1214 (1987). https://doi.org/10.1038/nbt1187-1207
