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Open Access Highly Accessed Research article

Bio-crude transcriptomics: Gene discovery and metabolic network reconstruction for the biosynthesis of the terpenome of the hydrocarbon oil-producing green alga, Botryococcus braunii race B (Showa)*

István Molnár12*, David Lopez3, Jennifer H Wisecaver4, Timothy P Devarenne5, Taylor L Weiss5, Matteo Pellegrini3 and Jeremiah D Hackett24

Author Affiliations

1 Natural Products Center, School of Natural Resources and the Environment, The University of Arizona, 250 E. Valencia Rd, Tucson, AZ, 85739, USA

2 Bio5 Institute, The University of Arizona, 1657 E. Helen St, Tucson, AZ, 85721, USA

3 Department of Molecular, Cell and Developmental Biology, University of California Los Angeles, P. O. Box 951606, Los Angeles, CA, 90095, USA

4 Department of Ecology and Evolutionary Biology, The University of Arizona, 1041 E. Lowell St, Tucson, AZ, 85721, USA

5 Department of Biochemistry and Biophysics, Texas A&M University, 2128 TAMU, College Station, TX, 77843, USA

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BMC Genomics 2012, 13:576  doi:10.1186/1471-2164-13-576

Published: 30 October 2012

Abstract

Background

Microalgae hold promise for yielding a biofuel feedstock that is sustainable, carbon-neutral, distributed, and only minimally disruptive for the production of food and feed by traditional agriculture. Amongst oleaginous eukaryotic algae, the B race of Botryococcus braunii is unique in that it produces large amounts of liquid hydrocarbons of terpenoid origin. These are comparable to fossil crude oil, and are sequestered outside the cells in a communal extracellular polymeric matrix material. Biosynthetic engineering of terpenoid bio-crude production requires identification of genes and reconstruction of metabolic pathways responsible for production of both hydrocarbons and other metabolites of the alga that compete for photosynthetic carbon and energy.

Results

A de novo assembly of 1,334,609 next-generation pyrosequencing reads form the Showa strain of the B race of B. braunii yielded a transcriptomic database of 46,422 contigs with an average length of 756 bp. Contigs were annotated with pathway, ontology, and protein domain identifiers. Manual curation allowed the reconstruction of pathways that produce terpenoid liquid hydrocarbons from primary metabolites, and pathways that divert photosynthetic carbon into tetraterpenoid carotenoids, diterpenoids, and the prenyl chains of meroterpenoid quinones and chlorophyll. Inventories of machine-assembled contigs are also presented for reconstructed pathways for the biosynthesis of competing storage compounds including triacylglycerol and starch. Regeneration of S-adenosylmethionine, and the extracellular localization of the hydrocarbon oils by active transport and possibly autophagy are also investigated.

Conclusions

The construction of an annotated transcriptomic database, publicly available in a web-based data depository and annotation tool, provides a foundation for metabolic pathway and network reconstruction, and facilitates further omics studies in the absence of a genome sequence for the Showa strain of B. braunii, race B. Further, the transcriptome database empowers future biosynthetic engineering approaches for strain improvement and the transfer of desirable traits to heterologous hosts.

Keywords:
Biofuel; Terpene biosynthesis; Fatty acid biosynthesis; Triacylglycerol biosynthesis; Starch biosynthesis; ABC transporter; Autophagy; Transcriptome; Botryococcus braunii; Botryococcene