Isolation and characterization of a gene from Streptomyces sp strain C5 that confers the ability to convert daunomycin to doxorubicin on Streptomyces lividans TK24

DNA sequence analysis of a region of the Streptomyces sp. strain C5 daunomycin biosynthesis gene cluster, located between the daunomycin polyketide biosynthesis gene cluster and a durl (transcriptional activator) homolog, revealed the presence of a gene encoding a P-450-like enzyme with a deduced M(r) of 46,096. Expression of this gene, named herein doxA, in Streptomyces lividans TK24 resulted in in vivo bioconversion of daunomycin to doxorubicin. DoxA showed specificity for only daunomycin and 13-dihydrodaunomycin, both of which were converted to doxorubicin. Daunomycinone (daunomycin aglycone), carminomycin, 13-dihydrocarminomycin, idarubicin, and aklavin were not apparent substrates for DoxA. In vector controls or in vectors in which doxA was poorly expressed, S. lividans catalyzed the reduction of daunomycin and other 13-oxo-anthracyclines and -anthracyclinones to their 13-dihydro homologs.