Functional analysis of the validamycin biosynthetic gene cluster and engineered production of validoxylamine A

Chem Biol. 2006 Apr;13(4):387-97. doi: 10.1016/j.chembiol.2006.02.002.

Abstract

A 45 kb DNA sequencing analysis from Streptomyces hygroscopicus 5008 involved in validamycin A (VAL-A) biosynthesis revealed 16 structural genes, 2 regulatory genes, 5 genes related transport, transposition/integration or tellurium resistance; another 4 genes had no obvious identity. The VAL-A biosynthetic pathway was proposed, with assignment of the required genetic functions confined to the sequenced region. A cluster of eight reassembled genes was found to support VAL-A synthesis in a heterologous host, S. lividans 1326. In vivo inactivation of the putative glycosyltransferase gene (valG) abolished the final attachment of glucose for VAL production and resulted in accumulation of the VAL-A precursor, validoxylamine, while the normal production of VAL-A could be restored by complementation with valG. The role of valG in the glycosylation of validoxylamine to VAL-A was demonstrated in vitro by enzymatic assay.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • DNA, Bacterial / genetics
  • Gene Targeting
  • Genes, Bacterial
  • Genes, Regulator
  • Genetic Complementation Test
  • Genetic Engineering
  • Glycosylation
  • Glycosyltransferases / genetics
  • Glycosyltransferases / metabolism
  • Inositol / analogs & derivatives*
  • Inositol / biosynthesis
  • Inositol / genetics
  • Molecular Sequence Data
  • Molecular Structure
  • Multigene Family
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Streptomyces / genetics*
  • Streptomyces / metabolism*
  • Streptomyces lividans / genetics
  • Streptomyces lividans / metabolism

Substances

  • DNA, Bacterial
  • Recombinant Proteins
  • validamycin A
  • validoxylamine A
  • Inositol
  • Glycosyltransferases

Associated data

  • GENBANK/AY753181