Abstract

Botryococcus braunii is a promising microalga for the production of biofuels and other chemicals because of its high content of internal lipids and external hydrocarbons. However, due to the very thick cell wall of B. braunii, traditional chemical/physical downstream processing very often is not as effective as expected and requires high amounts of energy. In this cases, the application of two-phase aqueous-organic solvent systems could be an alternative to cultivate microalgae allowing for a simultaneous extraction of the valuable compounds without significant negative effects on cell growth. Two-phase systems have been applied before, however, there are no studies so far on the mechanisms used by microalgae to survive in contact with solvents present as a second-phase. In this study, the effects of the solvents limonene, n-decane and n-decanol on growth of the microalga B. braunii as well as the adaptive cell response in terms of their phospholipid fatty acid contents were analized. A concentration-dependent negative effect of all three solvents on cell growth was observed. Effects were accompanied by changes of the membrane fatty acid composition of the alga as manifested by a decrease of the unsaturation . In addition, an association was found between the solvent hydrophobicity (given as log octanol–water partition coefficient (text {P}_{O-W}) values) and their toxic effects, whereby n-decanol and n-decane emerged as the most and least toxic solvent respectively. Among the tested solvents, the latter promises to be the most suitable for a two-phase extraction system.

Highlights

  • Botryococcus braunii is a microalga that can be found in fresh, brackish, and saline water all around the world (Aaronson et al 1983)

  • Cell growth The effect of three solvents of different log PO−W on B. braunii growth was measured (Fig. 1). n-decanol was found to be the most toxic solvent tested, resulting in a lower cell concentration compared to n-decane and limonene at quasi identical solvent concentrations (p-value < 0.01)

  • Limonene presents a monotonic ascending trend for C16:0 and the opposite for C18:1, for increasing solvent concentrations. These changes in fatty acid profile were reflected by different unsaturation index (UI), showing differences in response to both the solvent type and solvent concentration, suggesting predominance of saturated fatty acids and those fatty acids with one double bond

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Summary

Introduction

Botryococcus braunii is a microalga that can be found in fresh, brackish, and saline water all around the world (Aaronson et al 1983). For extracting biotechnologically valuable products from microorganisms generally two different methods are used: (i) intensive extraction from harvested biomass (Cooney et al 2009; Kumar et al 2015) and (ii) continuous extraction in a two-phase aqueous-organic solvent system, during ongoing microbial growth (Kleinegris et al 2011). This second approach has been used to extract valuable compounds such as carotenoids, lipids, and hydrocarbons from microalgae maintaining, whereby cell growth is, at least partially, maintained (Hejazi and Wijffels 2004; Sim et al 2001; Zhang et al 2011a).

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