The performance of microwaves irradiation (MAE and VMAE) to extract pigments from two marine microalgae was compared to conventional processes (cold and hot soaking and ultrasound-assisted extraction). Pigments were quantified by RP-HPLC and extraction performance was assessed regarding rapidity, reproducibility and extraction yields. Scanning electron microscopy was used at all extraction steps to assess the impact of the process on microalgal cell integrity. Freeze-drying and pigments extraction preserved microalgae cell integrity (except sonication) and evoked agglutination in superposed cells layers. All processes performed on Dunaliella tertiolecta (chlorophyte) lead to rapid pigments extraction, and equivalent pigments extraction yields, the absence of frustule allowing immediate solvent penetration in microalgae cells. In contrast, presence of the frustule in the diatom Cylindrotheca closterium (bacillariophyte) constituted a mechanical barrier to pigment extraction. MAE was identified as the best extraction process for CC pigments as it combined rapidity, reproducibility, homogeneous heating and high extraction yields.
In the present study, microwave-assisted extraction was first employed to extract the phycobiliproteins of Porphyridium purpureum (Pp). Freeze-dried Pp cells were subjected to microwave-assisted extraction (MAE) to extract phycoerythin (PE), phycocyanin (PC), and allophycocyanin (APC). MAE combined reproducibility and high extraction yields and allowed a 180- to 1,080-fold reduction of the extraction time compared to a conventional soaking process. The maximal PE extraction yield was obtained after 10-s MAE at 40 °C, and PE was thermally damaged at temperatures higher than 40 °C. In contrast, a flash irradiation for 10 s at 100 °C was the best process to efficiently extract PC and APC, as it combined a high temperature necessary to extract them from the thylakoid membrane to a short exposure to thermal denaturation. The extraction order of the three phycobiliproteins was coherent with the structure of Pp phycobilisomes. Moreover, the absorption and fluorescence properties of MAE extracted phycobiliproteins were stable for several months after the microwave treatment. Scanning electron microscopy indicated that MAE at 100 °C induced major changes in the Pp cell morphology, including fusion of the exopolysaccharidic cell walls and cytoplasmic membranes of adjacent cells. As a conclusion, MAE is a fast and high yield process efficient to extract and pre-purify phycobiliproteins, even from microalgae containing a thick exopolysaccharidic cell wall.
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