Properties and substrate specificity of RppA, a chalcone synthase-related polyketide synthase in Streptomyces griseus

J Biol Chem. 2002 Feb 15;277(7):4628-35. doi: 10.1074/jbc.M110357200. Epub 2001 Nov 26.

Abstract

RppA, a chalcone synthase-related polyketide synthase (type III polyketide synthase) in the bacterium Streptomyces griseus, catalyzes the formation of 1,3,6,8-tetrahydroxynaphthalene (THN) from five molecules of malonyl-CoA. The K(m) value for malonyl-CoA and the k(cat) value for THN synthesis were determined to be 0.93 +/- 0.1 microm and 0.77 +/- 0.04 min(-1), respectively. RppA accepted aliphatic acyl-CoAs with the carbon lengths from C(4) to C(8) as starter substrates and catalyzed sequential condensation of malonyl-CoA to yield alpha-pyrones and phloroglucinols. In addition, RppA yielded a hexaketide, 4-hydroxy-6-(2',4',6'-trioxotridecyl)-2-pyrone, from octanoyl-CoA and five molecules of malonyl-CoA, suggesting that the size of the active site cavity of RppA is larger than any other chalcone synthase-related enzymes found so far in plants and bacteria. RppA was also found to synthesize a C-methylated pyrone, 3,6-dimethyl-4-hydroxy-2-pyrone, by using acetoacetyl-CoA as the starter and methylmalonyl-CoA as an extender. Thus, the broad substrate specificity of RppA yields a wide variety of products.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acyltransferases / chemistry*
  • Acyltransferases / metabolism*
  • Chromatography, High Pressure Liquid
  • Indicators and Reagents / chemistry
  • Kinetics
  • Models, Chemical
  • Multienzyme Complexes / chemistry*
  • Naphthols / chemistry
  • Phloroglucinol / chemistry
  • Spectrophotometry
  • Streptomyces griseus / enzymology*
  • Substrate Specificity
  • Time Factors

Substances

  • 1,3,6,8-tetrahydroxynaphthalene
  • Indicators and Reagents
  • Multienzyme Complexes
  • Naphthols
  • Phloroglucinol
  • Acyltransferases
  • flavanone synthetase