Phycocyanin is a blue colored pigment, synthesized by several species of cyanobacteria and red algae. Besides the application as a food-colorant, the pigmented protein is of high interest as a pharmaceutically and nutritionally valuable compound. Since cyanobacteria-derived phycocyanin is thermolabile, red algae that are adapted to high temperatures are an interesting source for phycocyanin extraction. Still, the extraction of high quality phycocyanin from red algae is challenging due to the strong and rigid cell wall. Since standard techniques show low yields, alternative methods are needed. Recently, spark discharges have been shown to gently disintegrate microalgae and thereby enable the efficient extraction of susceptible proteins. In this study, the applicability of spark discharges for phycocyanin extraction from the red alga Cyanidium caldarium was investigated. The efficiency of 30 min spark discharges was compared with standard treatment protocols, such as three times repeated freeze-thaw cycles, sonication, and pulsed electric fields. Input energy for all physical methods were kept constant at 11,880 J to ensure comparability. The obtained extracts were evaluated by photometric and fluorescent spectroscopy. Highest extraction yields were achieved with sonication (53 mg/g dry weight (dw)) and disintegration by spark discharges (4 mg/g dw) while neither freeze-thawing nor pulsed electric field disintegration proved effective. The protein analysis via LC-MS of the former two extracts revealed a comparable composition of phycobiliproteins. Despite the lower total concentration of phycocyanin after application of spark discharges, the purity in the raw extract was higher in comparison to the extract attained by sonication.
Microalgae possess a cell wall with remarkable physical and chemical strength, which has proved to be a bottleneck for extraction techniques. With the application of spark discharges instigated by 100-ns high voltage pulses, a novel method was established that was found to be effective and yet gentle for the extraction of sensitive, especially heat-sensitive, compounds. The spark, which was ignited directly in the submerged algae suspension, is characterised by several physical and chemical processes that might promote a successful disintegration of the cells. Among other characteristics, the conceivably most destructive mechanism, i.e. strong shockwaves, was examined with schlieren diagnostics to evaluate associated pressures. The results were compared with the tensile strength, determined by atomic force microscopy, of Chlorella vulgaris, which was chosen as model organism. A shockwave pressure of 500 MPa in the close vicinity of the discharge by far exceeded the elasticity modulus of the algae with 13.95 MPa, confirming the potential of mechanically breaking the cell wall. At the same time, bulk temperatures could be maintained close to room temperature by adjusting the operating parameters for the spark-application, hence, especially encouraging the extraction of thermally unstable intracellular substances.
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