Gene Replacement of dnmV in Daunorubicin Producer SIPI-DM

Abstract:

C4-keto reduction in the TDP-daunosamine pathway of daunorubicin biosynthesis was catalyzed by DnmV ketoreductase. Epidaunorubicin producer has been constructed previously by disruption of dnmV gene and integration of aveBIV originated from biosynthesis pathway of avermectin. In this work, dnrIJ and dnmZU were amplified from genome of SIPI-DM, a high-daunorubicin producer. Recombinant plasmid pYG838 was constructed with aveBIV insertion between these two genes, which used as homologous recombinant arms. After screening and validation, an epidaunorubicin producer with dnmV replacing by aveBIV which is genetically and metabolically stable was obtained. The lack of antibiotic resistant marker in chromosome of this mutant will be helpful for further gene disruption.