Engineered biosynthesis of bacteriochlorophyll b in Rhodobacter sphaeroides

Biochim Biophys Acta. 2014 Oct;1837(10):1611-6. doi: 10.1016/j.bbabio.2014.07.011. Epub 2014 Jul 21.

Abstract

Bacteriochlorophyll b has the most red-shifted absorbance maximum of all naturally occurring photopigments. It has a characteristic ethylidene group at the C8 position in place of the more common ethyl group, the product of a C8-vinyl reductase, which is carried by the majority of chlorophylls and bacteriochlorophylls used in photosynthesis. The subsequent and first step exclusive to bacteriochlorophyll biosynthesis, the reduction of the C7=C8 bond, is catalyzed by chlorophyllide oxidoreductase. It has been demonstrated that the enzyme from bacteriochlorophyll a-utilizing bacteria can catalyze the formation of compounds carrying an ethyl group at C8 from both ethyl- and vinyl-carrying substrates, indicating a surprising additional C8-vinyl reductase function, while the enzyme from organisms producing BChl b could only catalyze C7=C8 reduction with a vinyl substrate, but this product carried an ethylidene group at the C8 position. We have replaced the native chlorophyllide oxidoreductase-encoding genes of Rhodobacter sphaeroides with those from Blastochloris viridis, but the switch from bacteriochlorophyll a to b biosynthesis is only detected when the native conventional C8-vinyl reductase is absent. We propose a non-enzymatic mechanism for ethylidene group formation based on the absence of cellular C8-vinyl reductase activity.

Keywords: Bacteriochlorophyll; Chlorophyll; Chlorophyllide oxidoreductase; Pathway engineering; Photosynthesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteriochlorophylls / biosynthesis*
  • Base Sequence
  • Biocatalysis
  • Chromatography, High Pressure Liquid
  • DNA Primers
  • Genes, Bacterial
  • Pigments, Biological / isolation & purification
  • Polymerase Chain Reaction
  • Rhodobacter sphaeroides / genetics
  • Rhodobacter sphaeroides / metabolism*

Substances

  • Bacteriochlorophylls
  • DNA Primers
  • Pigments, Biological
  • bacteriochlorophyll b