Anchoring a plant cytochrome P450 via PsaM to the thylakoids in Synechococcus sp. PCC 7002: evidence for light-driven biosynthesis

• Published in: PloS one 2014-07, Vol.9 (7), p.e102184-e102184
• Main Authors: Lassen, Lærke Münter, Nielsen, Agnieszka Zygadlo, Olsen, Carl Erik, Bialek, Wojciech, Jensen, Kenneth, Møller, Birger Lindberg, Jensen, Poul Erik
• Format: Article
• Language: English
• Subjects: Alternative energy sources; Anchoring; Biodiesel fuels; Biological activity; Biology and Life Sciences; Biosynthesis; Biotechnology; Catalysis; Chemical synthesis; Coupling (molecular); Cyanobacteria; Cytochrome; Cytochrome P-450 Enzyme System - genetics; Cytochrome P-450 Enzyme System - metabolism; Cytochrome P450; Electron transport; Electron transport chain; Endoplasmic reticulum; Engineering; Environmental science; Enzyme Activation; Enzymes; Gene Order; Homologous Recombination; Hydrogen; Light; Membranes; Metabolism; Metabolites; Nanoparticles; Photocatalysis; Photosynthesis; Photosystem I; Photosystem I Protein Complex - chemistry; Photosystem I Protein Complex - genetics; Photosystem I Protein Complex - metabolism; Phytochemistry; Plant sciences; Plants (botany); Protein Transport; Proteins; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Sorghum; Sorghum bicolor; Synechococcus - genetics; Synechococcus - metabolism; Synechocystis; Synthetic biology; Thylakoids; Thylakoids - metabolism; Transformation, Bacterial
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0102184

Plants produce an immense variety of specialized metabolites, many of which are of high value as their bioactive properties make them useful as for instance pharmaceuticals. The compounds are often produced at low levels in the plant, and due to their complex structures, chemical synthesis may not be feasible. Here, we take advantage of the reducing equivalents generated in photosynthesis in developing an approach for producing plant bioactive natural compounds in a photosynthetic microorganism by functionally coupling a biosynthetic enzyme to photosystem I. This enables driving of the enzymatic reactions with electrons extracted from the photosynthetic electron transport chain. As a proof of concept, we have genetically fused the soluble catalytic domain of the cytochrome P450 CYP79A1, originating from the endoplasmic reticulum membranes of Sorghum bicolor, to a photosystem I subunit in the cyanobacterium Synechococcus sp. PCC 7002, thereby targeting it to the thylakoids. The engineered enzyme showed light-driven activity both in vivo and in vitro, demonstrating the possibility to achieve light-driven biosynthesis of high-value plant specialized metabolites in cyanobacteria.