In vivo characterization of NcsB3 to establish the complete biosynthesis of the naphthoic acid moiety of the neocarzinostatin chromophore

FEMS Microbiol Lett. 2010 Oct;311(2):119-25. doi: 10.1111/j.1574-6968.2010.02075.x. Epub 2010 Aug 23.

Abstract

Neocarzinostatin (NCS) is an enediyne antibiotic produced by Streptomyces carzinostaticus. The NCS chromophore consists of an enediyne core, a sugar moiety, and a naphthoic acid (NA) moiety. The latter plays a key role in binding the NCS chromophore to its apoprotein to protect and stabilize the bioactive NCS chromophore. In this study, we expressed three genes: ncsB (naphthoic acid synthase), ncsB3 (P450 hydroxylase), and ncsB1 (O-methyltransferase), in Streptomyces lividans TK24. The three genes were sufficient to produce 2-hydroxy-7-methoxy-5-methyl-1-naphthoic acid. Production was analyzed and confirmed by LC-MS and nuclear magnetic resonance. Here, we report the functional characterization of ncsB3 and thereby elucidate the complete biosynthetic pathway of NA moiety of the NCS chromophore.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Bacterial Agents / biosynthesis*
  • Anti-Bacterial Agents / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Biosynthetic Pathways
  • Carboxylic Acids / chemistry
  • Carboxylic Acids / metabolism*
  • Cytochrome P-450 Enzyme System / genetics
  • Cytochrome P-450 Enzyme System / metabolism*
  • Gene Expression Regulation, Bacterial
  • Molecular Structure
  • Naphthalenes / chemistry
  • Naphthalenes / metabolism*
  • Streptomyces / chemistry
  • Streptomyces / enzymology
  • Streptomyces / metabolism*
  • Zinostatin / analogs & derivatives
  • Zinostatin / biosynthesis
  • Zinostatin / chemistry

Substances

  • Anti-Bacterial Agents
  • Bacterial Proteins
  • Carboxylic Acids
  • Naphthalenes
  • 1-naphthoic acid
  • neocarzinostatin chromophore
  • Zinostatin
  • Cytochrome P-450 Enzyme System