CHARACTERIZATION OF THE CEFF GENE OF NOCARDIA LACTAMDURANS ENCODING A3'-METHYLCEPHEM HYDROXYLASE DIFFERENT FROM THE 7-CEPHEM HYDROXYLASE

The cefF gene of Nocardia lactamdurans, encoding a functional 2-oxoglu tarate-dependent 3'-methylcephem hydroxylase (deacetoxycephalosporin C hydroxylase) has been found to be closely linked to the pcbC gene in the cephamycin C gene cluster. The open-reading frame is 933 bp long a nd could encode a protein of M(r) 34366. Introduction of ceJF in the c ephamycin-non-producer Streptomyces lividans conferred 3'-methylcephem -hydroxylating activity to the transformants but did not result in hyd roxylation at carbon 7 of cephamycin. No 3'-methylcephem hydroxylase a ctivity was observed when the ceJF gene was introduced in S. lividans in pIJ699 (a vector containing two transcriptional terminators that pr event readthrough expression), which suggests that this gene lacks an independent promoter.