Manipulating Fatty Acid Biosynthesis in Microalgae for Biofuel through Protein-Protein Interactions

Microalgae are a promising feedstock for renewable fuels, and algal metabolic engineering can lead to crop improvement, thus accelerating the development of commercially viable biodiesel production from algae biomass. We demonstrate that protein-protein interactions between the fatty acid acyl carri...

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Veröffentlicht in:	PloS one Jg. 7; H. 9; S. e42949
Hauptverfasser:	Blatti, Jillian L., Beld, Joris, Behnke, Craig A., Mendez, Michael, Mayfield, Stephen P., Burkart, Michael D.
Format:	Journal Article
Sprache:	Englisch
Veröffentlicht:	United States Public Library of Science 13.09.2012 Public Library of Science (PLoS)
Schlagworte:	Acyl carrier protein Acyl Carrier Protein - chemistry Acyl Carrier Protein - metabolism Algae Alternative fuels Amino Acid Sequence Amino acids Antibiotics Aquatic microorganisms BASIC BIOLOGICAL SCIENCES Biochemistry Biodiesel fuels Biofuels Bioinformatics Biology Biomass Biomass energy Biosynthesis Brassica napus Chemistry Chlamydomonas Chlamydomonas reinhardtii Chlamydomonas reinhardtii - genetics Chlamydomonas reinhardtii - metabolism Chloroplasts Chloroplasts - metabolism Computer simulation Crop improvement Cross-linking Crosslinking Diesel fuels Docking Engineering Enzymes Fatty acids Fatty Acids - biosynthesis Fuels Gene Expression Genetic engineering Hydrolysis Metabolic engineering Metabolism Microalgae Microalgae - genetics Microalgae - metabolism Molecular Docking Simulation Molecular Sequence Data Neural networks Phaeodactylum tricornutum Plant Proteins - metabolism Plants Plants, Genetically Modified Polymer crosslinking Protein Binding Protein Conformation Protein interaction Protein-protein interactions Proteins Proteins - metabolism Renewable fuels Science & Technology - Other Topics Sequence Alignment Substrate Specificity Thioesterase Thiolester Hydrolases - chemistry Thiolester Hydrolases - metabolism La Jolla California United States > US California
ISSN:	1932-6203, 1932-6203
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Zusammenfassung:	Microalgae are a promising feedstock for renewable fuels, and algal metabolic engineering can lead to crop improvement, thus accelerating the development of commercially viable biodiesel production from algae biomass. We demonstrate that protein-protein interactions between the fatty acid acyl carrier protein (ACP) and thioesterase (TE) govern fatty acid hydrolysis within the algal chloroplast. Using green microalga Chlamydomonas reinhardtii (Cr) as a model, a structural simulation of docking CrACP to CrTE identifies a protein-protein recognition surface between the two domains. A virtual screen reveals plant TEs with similar in silico binding to CrACP. Employing an activity-based crosslinking probe designed to selectively trap transient protein-protein interactions between the TE and ACP, we demonstrate in vitro that CrTE must functionally interact with CrACP to release fatty acids, while TEs of vascular plants show no mechanistic crosslinking to CrACP. This is recapitulated in vivo, where overproduction of the endogenous CrTE increased levels of short-chain fatty acids and engineering plant TEs into the C. reinhardtii chloroplast did not alter the fatty acid profile. These findings highlight the critical role of protein-protein interactions in manipulating fatty acid biosynthesis for algae biofuel engineering as illuminated by activity-based probes.
Bibliographie:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 14 content type line 23 USDOE Office of Energy Efficiency and Renewable Energy (EERE) National Science Foundation (NSF) California Energy Commission EE0003373; 0742551; CILMSF 500-10-039 Conceived and designed the experiments: MDB JLB. Performed the experiments: JB JLB. Analyzed the data: MDB MM SPM CB JB JLB. Contributed reagents/materials/analysis tools: MM CB. Wrote the paper: MDB JLB. Competing Interests: MM and CB are employees of Sapphire Energy, which has competing interest in the biofuel arena. SPM was a founder of Sapphire and thus owns significant shares in the company. The research in this manuscript is performed through a US Department of Energy-funded collaboration between Sapphire Energy and UCSD. This does not alter the authors' adherence to all the PLoS ONE policies on sharing data and materials. Therefore, the rest of the authors declare no competing interest.
ISSN:	1932-6203 1932-6203
DOI:	10.1371/journal.pone.0042949