Streptomyces peucetius daunorubicin biosynthesis gene, dnrF: sequence and heterologous expression

Microbiology. 1995 Apr;141 ( Pt 4):1007-16. doi: 10.1099/13500872-141-4-1007.

Abstract

The dnrF gene, responsible for conversion of aklavinone to epsilon-rhodomycinone via C-11 hydroxylation, was mapped in the daunorubicin (Dnr) gene cluster of Streptomyces peucetius ATCC 29050, close to drrAB, one of the anthracycline-resistance genes. The dnrF gene was sequenced and should encode a protein of 489 amino acids with a molecular mass of 52 kDa. The deduced DnrF protein shows significant similarities with bacterial FAD- and NADPH-dependent hydroxylases either required to introduce hydroxyl groups into polycyclic aromatic polyketide antibiotics or involved in catabolism of aromatic compounds. Heterologous expression of dnrF in Streptomyces lividans TK23 and in Escherichia coli demonstrated that the gene encodes a NADPH-dependent hydroxylase catalysing the hydroxylation of aklavinone to yield epsilon-rhodomycinone. The enzyme is inactive on anthracyclines glycosylated at position C-7 and its activity decreases to a different extent with other substrate modifications, indicating that DnrF has a significant substrate specificity.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Aryl Hydrocarbon Hydroxylases / genetics
  • Aryl Hydrocarbon Hydroxylases / metabolism
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Base Sequence
  • Chromosome Mapping
  • Cloning, Molecular
  • DNA Primers / genetics
  • DNA, Bacterial / genetics
  • Daunorubicin / biosynthesis*
  • Escherichia coli / genetics
  • Gene Expression Regulation, Bacterial
  • Genes, Bacterial*
  • Molecular Sequence Data
  • Open Reading Frames
  • Sequence Homology, Amino Acid
  • Streptomyces / genetics*
  • Streptomyces / metabolism
  • Substrate Specificity

Substances

  • Bacterial Proteins
  • DNA Primers
  • DNA, Bacterial
  • aklavinone 11-hydroxylase
  • Aryl Hydrocarbon Hydroxylases
  • Daunorubicin

Associated data

  • GENBANK/U18082