# Mining Thermophile Photosynthesis Genes: A Synthetic Operon Expressing Chloroflexota Species Reaction Center Genes in Rhodobacter sphaeroides

## TL;DR

Scientists used a lab strain to express photosynthesis genes from hot spring microbes, revealing new insights into high-temperature photosynthesis.

## Contribution

A synthetic operon enabled the expression of Chloroflexota photosynthesis genes in Rhodobacter sphaeroides, enabling functional studies.

## Key findings

- A functional reaction center complex was produced from Chloroflexota genes in Rhodobacter sphaeroides.
- Metagenomic analysis identified diverse anoxygenic type II reaction center sequences from high-temperature springs.
- The approach provides a resource for studying photosynthesis in thermophilic environments and potential biotech applications.

## Abstract

Photosynthesis is the foundation of the vast majority of life systems, and is therefore the most important bioenergetic process on earth. The greatest diversity of photosynthetic systems is found in microorganisms. However, our understanding of the biophysical and biochemical processes that transduce light into chemical energy is derived from a relatively small subset of proteins from microbes that are amenable to cultivation, in contrast to the huge number of predicted proteins that catalyze the initial photochemical reactions deposited in databases, such as from metagenomics. We describe the use of a Rhodobacter sphaeroides laboratory strain for the expression of heterologous photosynthesis genes to demonstrate the feasibility of mining this resource, focusing on hot spring Chloroflexota gene sequences. Using a synthetic operon of genes, we produced a photochemically active complex of reaction center proteins in our biological system. We also present bioinformatic analyses of anoxygenic type II reaction center sequences from metagenomic samples collected from hot (42–90 °C) springs available through the JGI IMG database, to generate a resource of diverse sequences that are potentially adapted to photosynthesis at such temperatures. These data provide a view into the natural diversity of anoxygenic photosynthesis, through a lens focused on high-temperature environments. The approach we took to express such genes can be applied for potential biotechnology purposes as well as for studies of fundamental catalytic properties of these heretofore inaccessible protein complexes.