Expression of holo-proteorhodopsin in Synechocystis sp. PCC 6803

Metab Eng. 2016 May:35:83-94. doi: 10.1016/j.ymben.2016.02.001. Epub 2016 Feb 8.

Abstract

Retinal-based photosynthesis may contribute to the free energy conversion needed for growth of an organism carrying out oxygenic photosynthesis, like a cyanobacterium. After optimization, this may even enhance the overall efficiency of phototrophic growth of such organisms in sustainability applications. As a first step towards this, we here report on functional expression of the archetype proteorhodopsin in Synechocystis sp. PCC 6803. Upon use of the moderate-strength psbA2 promoter, holo-proteorhodopsin is expressed in this cyanobacterium, at a level of up to 10(5) molecules per cell, presumably in a hexameric quaternary structure, and with approximately equal distribution (on a protein-content basis) over the thylakoid and the cytoplasmic membrane fraction. These results also demonstrate that Synechocystis sp. PCC 6803 has the capacity to synthesize all-trans-retinal. Expressing a substantial amount of a heterologous opsin membrane protein causes a substantial growth retardation Synechocystis, as is clear from a strain expressing PROPS, a non-pumping mutant derivative of proteorhodopsin. Relative to this latter strain, proteorhodopsin expression, however, measurably stimulates its growth.

Keywords: All-trans retinal; Apo- and holo-protein; Cytoplasmic membrane; Proton motive force; Proton pump; Thylakoid membrane.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Gene Expression*
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics
  • Rhodopsins, Microbial / biosynthesis*
  • Rhodopsins, Microbial / genetics
  • Synechocystis / genetics
  • Synechocystis / metabolism*

Substances

  • Recombinant Proteins
  • Rhodopsins, Microbial
  • proteorhodopsin