Development of a cooperative two-factor adaptive-evolution method to enhance lipid production and prevent lipid peroxidation in Schizochytrium sp.

Abstract

BackgroundSchizochytrium sp. is a marine microalga with great potential as a promising sustainable source of lipids rich in docosahexaenoic acid (DHA). This organism’s lipid accumulation machinery can be induced by various stress conditions, but this stress induction usually comes at the expense of lower biomass in industrial fermentations. Moreover, oxidative damage induced by various environmental stresses can result in the peroxidation of lipids, and especially polyunsaturated fatty acids, which causes unstable DHA production, but is often ignored in fermentation processes. Therefore, it is urgent to develop new production strains that not only have a high DHA production capacity, but also possess strong antioxidant defenses.ResultsAdaptive laboratory evolution (ALE) is an effective method for the development of beneficial phenotypes in industrial microorganisms. Here, a novel cooperative two-factor ALE strategy based on concomitant low temperature and high salinity was applied to improve the production capacity of Schizochytrium sp. Low-temperature conditions were used to improve the DHA content, and high salinity was applied to stimulate lipid accumulation and enhance the antioxidative defense systems of Schizochytrium sp. After 30 adaptation cycles, a maximal cell dry weight of 126.4 g/L and DHA yield of 38.12 g/L were obtained in the endpoint strain ALE-TF30, which was 27.42 and 57.52% higher than parental strain, respectively. Moreover, the fact that ALE-TF30 had the lowest concentrations of reactive oxygen species and malondialdehyde among all strains indicated that lipid peroxidation was greatly suppressed by the evolutionary process. Accordingly, the ALE-TF30 strain exhibited an overall increase of gene expression levels of antioxidant enzymes and polyketide synthases compared to the parental strain.ConclusionThis study provides important clues on how to overcome the negative effects of lipid peroxidation on DHA production in Schizochytrium sp. Taken together, the cooperative two-factor ALE process can not only increase the accumulation of lipids rich in DHA, but also prevent the loss of produced lipid caused by lipid peroxidation. The strategy proposed here may provide a new and alternative direction for the industrial cultivation of oil-producing microalgae.Electronic supplementary materialThe online version of this article (10.1186/s13068-018-1065-4) contains supplementary material, which is available to authorized users.

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“…Thus, the development of new strains that simultaneously produce high amounts of lipids and show antioxidant robustness are desired. An adaptive laboratory evolution (ALE) was constructed by Sun et al [27] to enhance the Shizochytrium DHA production. The authors used two different approaches for the ALE: low temperatures and high salinities to improve the antioxidant resistance of the microalga.…”

“…Most of the recent works reporting lipid/DHA production from heterotrophic microalgae use traditional methods to monitor the microalgae growth during the culture development, such as optical density, dry cell weight and cell counting [19,[22][23][24][25][26][27][28][29][30][31][32][33][34][35][36][37][38][39][40]. However, these techniques provide no information on cell physiological states.…”