Characterization of chlorophyll a and bacteriochlorophyll a synthases by heterologous expression in Escherichia coli

J Biol Chem. 1997 Apr 11;272(15):9671-6. doi: 10.1074/jbc.272.15.9671.

Abstract

Genes coding for putative chlorophyll a synthase (chlG) from Synechocystis sp. PCC 6803 and bacteriochlorophyll a synthase (bchG) from Rhodobacter capsulatus were amplified by the polymerase chain reaction and cloned into T7 RNA polymerase-based expression plasmids. In vitro enzymatic assays indicated that heterologous expression of the chlG and bchG gene products in Escherichia coli conferred chlorophyll a and bacteriochlorophyll a synthase activity, respectively. Chlorophyll a synthase utilized chlorophyllide a, but not bacteriochlorophyllide a, as a substrate, whereas bacteriochlorophyll a synthase utilized bacteriochlorophyllide a, but not chlorophyllide a. Both enzymes were also observed to exhibit a marked preference for phytyl diphosphate over geranylgeranyl diphosphate.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Bacteriochlorophylls / biosynthesis*
  • Carbon-Oxygen Ligases*
  • Chlorophyll / biosynthesis*
  • Chlorophyll A
  • Chromatography, High Pressure Liquid
  • Escherichia coli
  • Ligases / genetics*
  • Ligases / metabolism
  • Pigments, Biological / biosynthesis*
  • Spectrophotometry, Ultraviolet

Substances

  • Bacteriochlorophylls
  • Pigments, Biological
  • Chlorophyll
  • Ligases
  • Carbon-Oxygen Ligases
  • bacteriochlorophyll a synthase
  • chlorophyll synthetase
  • Chlorophyll A