Acyclic isoprenoid biomarkers and evolution of biosynthetic pathways in green microalgae of the genus Botryococcus

Acyclic isoprenoids were among the earliest lipids on Earth and today > 30,000 isoprenoid-derived compounds are known, testifying to the remarkable diversification in isoprenoid biosynthetic pathways over time. Many of the isoprenoids have proven to be useful biomarkers in geochemical studies and evidence from sedimentary studies has the potential to provide a timeline for the evolution of different types of isoprenoid biosynthesis. A single green microalgal species termed Botryococcus braunii has been recognised as a major contributor of organic matter to sediments as old as the Precambrian. Modern studies of the taxonomy of B. braunii using molecular biology techniques have shown that the major clades identified from 18S rRNA gene sequences correspond to races A, B, L and S, which are defined on the basis of their hydrocarbon composition. The biosynthetic pathways by which C₃₀–C₃₇ botryococcenes are produced by the B race have now been shown to be due to a duplication of the squalene synthase gene, followed by subsequent changes to the genes, such that one pathway leads to the production of the C₃₀ botryococcene and the other to squalene. Both products are then methylated to produce botryococcenes and methylated squalenes having higher carbon numbers. The mode of biosynthesis of lycopadiene in race L is unclear, but may involve coupling of two C₂₀ phytyl diphosphates. From an examination of the geological record of botryococcenes and lycopadiene it seems likely that these pathways probably evolved early in the Eocene (ca. 55 Ma) and thus are more recently evolved than the genes for highly branched isoprenoid (HBI) alkenes first produced by diatoms about 92 Ma ago. Botryococcane produced from botryococcenes and a monoaromatic hydrocarbon produced from lycopadiene-related lipids, presumably under anoxic conditions, show promise as age diagnostic biomarkers. In view of these results, it seems likely that the Botryococcus species recorded in sediments predating the Eocene lacked the ability to produce botryococcenes or lycopadiene, but nonetheless still contained polymeric non-isoprenoid alkyl chains in the form of an algaenan which on preservation gave rise to a multitude of organic rich rocks.