Effects of sulfur and phosphorus concentration on the lipid accumulation and fatty acid profile in Chlorella vulgaris (Chlorophyta).

Nutrient deficiency induces a variety of cellular responses, including an increase in lipid accumulation in microalgae. Nitrogen starvation is the most studied deprivation. Here, we determine the effects of phosphorus and sulfur limitation on lipid accumulation in Chlorella vulgaris. A set of 9 experiments were performed, varying the initial concentration of these nutrients (set to 0, 50, and 100% of their original composition in Bold's basal medium). According to our results, the variation of P and S modified the specific growth rate, lag phase, and cell generation time. The ratio of 50%P and 0%S significantly increased the total lipid concentration. The fatty acid profile was dominated by C16:0, C18:0, and C18:1; a considerable increase in C20:5 was observed with 0%P and 50%S and 0%P and 100%S. Regarding neutral lipids, the response surface methodology (RSM) indicates that the maximum was observed when S was between 40 and 60% and P was between 95 and 100%. Therefore, the enhanced production of lipids caused by P and S limitation may contribute to the efficient oil production useful for algal biofuels.

Effect of the double bond conjugation on the vascular physiology and nitric oxide production of isomers of eicosapentaenoic and docosahexaenoic acids prepared from shark oil.

A collection of evidence suggests that conjugation of double bonds of eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids, omega-3 polyunsaturated fatty acids (n-3 PUFAs), increases their anticarcinogenic activity; however, the effect of such conjugation on vascular tone activity remains unknown. We propose that the mixture of conjugated PUFAs exerts higher vasorelaxation activity than the corresponding mixture of nonconjugated PUFAs. The vascular response to different concentrations of conjugated and nonconjugated isomers of EPA and DHA, among other fatty acids (FAs) naturally present in shark oil, and the role of nitric oxide (NO) as a vasorelaxant agent were investigated. Both conjugated EPA (CEPA) and conjugated DHA (CDHA) were prepared by alkaline isomerization of all PUFAs contained in shark oil. Different concentrations of conjugated and nonconjugated PUFAs were placed in contact with precontracted aortic rings of Wistar rats to assess their effect on vascular tone. All tested samples exerted a vasorelaxant effect. Compared to nonconjugated PUFAs, conjugated isomers exhibited an increase in the dilatation of the aortic rings (P<0.001) in a dose-dependent manner (P<0.001). In addition, nonconjugated PUFAs produced nitric oxide (NO) in a dose-dependent manner, while conjugated PUFAs did not, suggesting that their dilatation mechanism is not totally dependent on NO.

pH effects on the lipid and fatty acids accumulation in Chlamydomonas reinhardtii.

pH variations influence the delivery of essential nutrients and CO2 solubility, which impact algae metabolism. In this study the microalgal growth and chlorophyll, lipid, and fatty acids content; along with the expression of some genes implicated in the biosynthesis of lipids were examined in Chlamydomonas reinhardtii subjected to pH values of 7.0, 7.8, and 8.5. At pH 7.8 an increase in cell growth was observed with a significant accumulation of chlorophyll (1.75-fold) when compared with growth at pH 7, while at pH 8.5 a sharp decrease in both parameters was observed when compared with the other pH values tested. Lipid content increased 3.0 (14.81% of dry cell weight, dcw) and 2.3 times (11.43% dcw) at pH 7.8 and 8.5, respectively, when compared with the experiment at pH 7 (4.97% dcw). The compositions of major fatty acids in the strains growing at pH 7.0, 7.8, or 8.5 were 25.7, 28.0, and 32.1% for palmitic acid; 17.3, 14.7, and 25.7% for oleic acid; and 9.8, 12.1, and 4.6% for linoleic acid; respectively. qRT-PCR analysis showed that the transcripts of ß-carboxyltransferase, Acyl carrier protein 1, acyl-ACP thiolase 1, acyl-sn-glycerol-3-phosphate acyltransferase, and diacylglycerol acyl transferase isoform 3 were significantly induced at pH 7.8 when compared with the other two pH conditions. These results indicate that the induction of genes implicated in the early and final steps of lipid biosynthesis contributes to their accumulation in the stationary phase. Our research suggests that a pH of 7.8 might be ideal to maximize growth and lipid accumulation.

Effect of green and red light in lipid accumulation and transcriptional profile of genes implicated in lipid biosynthesis in Chlamydomonas reinhardtii.

Microalgae have the potential to accumulate triacylglycerols under different light spectra. In this work, Chlamydomonas reinhardtii was grown under white (400-700 nm), red (650 nm), and green (550 nm) lights. According to our results, red light (650 nm) has a positive effect in the microalgae growth and chlorophyll concentration. About the lipid content, the control culture (white light-illuminated) reached a 4.4% of dry cell weight (dcw), whereas the culture grown at 550 nm showed an increase of 1.35-fold in the lipids accumulation (5.96% dcw). Interestingly, the most significant accumulation was found in the culture grown at 650 nm (14.78% dcw) which means 3.36-fold higher with respect to the white light-illuminated culture. The most abundant fatty acids found in lipid extracts obtained from the cultures under different light wavelength were palmitic (C16: 0), oleic (C18: 1n9), stearidonic (C18: 4), and linoleic (C18: 2), which are useful in the biodiesel production. Changes in gene expression in response to different wavelength illuminations were assessed; however, an in-depth analysis of a larger number of genes involved in lipid biosynthesis is necessary to fully explain the highest accumulation of lipids in the culture grown under red light. This approach will be useful to find a sustainable source of lipids for biodiesel production. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1404-1411, 2016.

Over-expression of Dof-type transcription factor increases lipid production in Chlamydomonas reinhardtii.

The high demand for less polluting, newer, and cheaper fuel resources has increased the search of the most innovative options for the production of the so-called biofuels. Chlamydomonas reinhardtii is a photosynthetic unicellular algae with multiple biotechnological advantages such as easy handling in the laboratory, a simple scale-up to industrial levels, as well as a feasible genetic modification at nuclear and chloroplast levels. Besides, its fatty acids can be used to produce biofuels. Previous studies in plants have found that the over expression of DOF-type transcription factor genes increases the synthesis and the accumulation of total lipids in seeds. In this context, the over-expression of a DOF-type transcription factor in C. reinhardtii was applied as approach to increase the amount of lipids. The results indicate higher amounts (around 2-fold) of total lipids, which are mainly fatty acids, in the genetically C. reinhardtii modified strains when compared with the non-genetically modified strain. In order to elucidate the possible function of the introduced Dof-type transcription factor, we performed a transcription profile of 8 genes involved in fatty acid biosynthesis and 6 genes involved in glycerolipid biosynthesis, by quantitative real time (qRT-PCR). Differential expression profile was observed, which can explain the increase in lipid accumulation. However, these strains did not show notable changes in the fatty acid profile. This work represents an early effort in generating a strategy to increase fatty acids production in C. reinhardtii and their use in biofuel synthesis.