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

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 b...

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Bibliographic Details
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
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Summary: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.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0102184