PRODUCTION OF CEPHALOSPORIN INTERMEDIATES BY FEEDING ADIPIC ACID TO RECOMBINANT PENICILLIUM-CHRYSOGENUM STRAINS EXPRESSING RING EXPANSION ACTIVITY

We demonstrate a novel and efficient bioprocess for production of the cephalosporin intermediates, 7-aminocephalosporanic acid (7-ACA) or 7- amino deacetoxycephalosporanic acid (7-ADCA). The Streptomyces clavuli gerus expandase gene or the Cephalosporium acremonium expandase-hydrox ylase gene, with and without the acetyltransferase gene, were expresse d in a penicillin production strain of Penicillium chrysogenum. Growth of these transformants in media containing adipic acid as the side ch ain precursor resulted in efficient production of cephalosporins havin g an adipyl side chain, proving that adipyl-6-APA is a substrate for e ither enzyme in vivo. Strains expressing expandase produced adipyl-7-A DCA, whereas strains expressing expandase-hydroxylase produced both ad ipyl-7-ADCA and adipyl-7-ADAC (aminodeacetylcephalosporanic acid). Str ains expressing expandase-hydroxylase and acetyltransferase produced a dipyl-7-ADCA, adipyl-7-ADAC and adipyl-7-ACA. The adipyl side chain of these cephalosporins was easily removed with a Pseudomonas-derived am idase to yield the cephalosporin intermediates.