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Optimization of a novel lipid extraction process from microalgae.

ABSTRACT: Previous study found that the solvent extraction efficiency of lipid in microalgae could be greatly improved by washing algae cells before the second time extraction. Based on the "organic solvents-water-organic solvents" method, this research further studied the effect of four solvent systems (acetone, chloroform/methanol, chloroform/methanol/water, dichloromethane/methanol), two types of water treatment (vortex and ultrasonic), three water treatment time gradient (0 s, 30 s, 120 s) on the lipid extraction at three different microalgae growth stages (3rd day, 5th day, 9th day). The results show that the combination of water treatment type, treatment time and solvent is very important to the efficiency of lipid extraction. The total lipid extracted was generally increased by 10-30% after water treatment. Especially under the condition of 120 s vortex water treatment with dichloromethane/methanol as extraction solvent, the total lipid extracted increased by 61.14%. In addition, microalgae cells at different culture stages had different sensitivity to water treatment. In this study, under the combination of chloroform/methanol/water as extraction solvent and vortex water treatment for 120 s, the highest lipid yield was obtained on the ninth day of cell culture, which accounts 47.88% of the cell dry weight (478 mg/g cell dry weight). The changes of cell morphology and structure after water treatment were studied by scanning electron microscope, and it was found that water treatment could seriously destroy the cell membrane damaged by solvent, thus promoting the release of lipids. This study further optimizes the "solvent-water-solvent" lipid extraction method, which neither produces impurities nor damages the lipid quality, and can reduce the amount of organic solvent applied in the classical lipid extraction method with the same lipid yield, so it has a broad application prospect.

SUBMITTER: Ren X

PROVIDER: S-EPMC8511141 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BackgroundRecent studies of lipid extraction from microalgae have focused primarily on dewatered or dried samples, and the processes are simple with high lipid yield. Yet, the dewatering with drying step is energy intensive, which makes the energy input during the lipid production more than energy output from obtained lipid. Thus, exploring an extraction technique for just a thickened sample without the dewatering, drying and auxiliary operation (such as cell disruption) is very significant. Whereas lipid extraction from the thickened microalgae is complicated by the high water content involved, and traditional solvent, hence, cannot work well. Dimethyl ether (DME), a green solvent, featuring a high affinity for both water and organic compounds with an ability to penetrate the cell walls has the potential to achieve this goal.ResultsThis study investigated an energy-saving method for lipid extraction using DME as the solvent with an entrainer solution (ethanol and acetone) for flocculation-thickened microalgae. Extraction efficiency was evaluated in terms of extraction time, DME dosage, entrainer dosage, and ethanol:acetone ratio. Optimal extraction occurred after 30 min using 4.2 mL DME per 1 mL microalgae, with an entrainer dosage of 8% at 1:2 ethanol:acetone. Raw lipid yields and its lipid component (represented by fatty acid methyl ester) contents were compared against those of common extraction methods (Bligh and Dryer, and Soxhlet). Thermal gravimetry/differential thermal analysis, Fourier-transform infrared spectroscopy, and C/H/N elemental analyses were used to examine differences in lipids extracted using each of the evaluated methods. Considering influence of trace metals on biodiesel utilization, inductively coupled plasma mass spectrometry and inductively coupled plasma atomic emission spectroscopy analyses were used to quantify trace metals in the extracted raw lipids, which revealed relatively high concentrations of Mg, Na, K, and Fe.ConclusionsOur DME-based method recovered 26.4% of total raw lipids and 54.4% of total fatty acid methyl esters at first extraction with remnants being recovered by a 2nd extraction. In additional, the DME-based approach was more economical than other methods, because it enabled simultaneous dewatering with lipid extraction and no cell disruption was required. The trace metals of raw lipids indicated a purification demand in subsequent refining process.

Project description:BackgroundProtein N-glycosylation is initiated within the endoplasmic reticulum through the synthesis of a lipid-linked oligosaccharides (LLO) precursor. This precursor is then transferred en bloc on neo-synthesized proteins through the action of the oligosaccharyltransferase giving birth to glycoproteins. The N-linked glycans bore by the glycoproteins are then processed into oligomannosides prior to the exit of the glycoproteins from the endoplasmic reticulum and its entrance into the Golgi apparatus. In this compartment, the N-linked glycans are further maturated in complex type N-glycans. This process has been well studied in a lot of eukaryotes including higher plants. In contrast, little information regarding the LLO precursor and synthesis of N-linked glycans is available in microalgae.MethodsIn this report, a user-friendly extraction method combining microsomal enrichment and solvent extractions followed by purification steps is described. This strategy is aiming to extract LLO precursor from microalgae. Then, the oligosaccharide moiety released from the extracted LLO were analyzed by multistage tandem mass spectrometry in two models of microalgae namely the green microalgae, Chlamydomonas reinhardtii and the diatom, Phaeodactylum tricornutum.ResultsThe validity of the developed method was confirmed by the analysis of the oligosaccharide structures released from the LLO of two xylosyltransferase mutants of C. reinhardtii confirming that this green microalga synthesizes a linear Glc3Man5GlcNAc2 identical to the one of the wild-type cells. In contrast, the analysis of the oligosaccharide released from the LLO of the diatom P. tricornutum demonstrated for the first time a Glc2Man9GlcNAc2 structure.ConclusionThe method described in this article allows the fast, non-radioactive and reliable multistage tandem mass spectrometry characterization of oligosaccharides released from LLO of microalgae including the ones belonging to the Phaeodactylaceae and Chlorophyceae classes, respectively. The method is fully adaptable for extracting and characterizing the LLO oligosaccharide moiety from microalgae belonging to other phyla.

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Optimization of a novel lipid extraction process from microalgae.

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