In light of climate change, there is a growing interest for sustainable energy. Microalgae are a promising resource for biofuel production, although their industrial use is limited by the lack of effective harvesting techniques. Flocculation consists in the aggregation and adhesion of cells into flocs that can be more easily removed from water than individual cells. Although it is an efficient harvesting technique, contamination is a major issue as chemical flocculants are often used. An alternative is to use natural biopolymers flocculants, such as chitosan. Chitosan is a bio-based nontoxic polymer, which has been effectively used to harvest Chlorella vulgaris cells at pH lower than its pKa (6.5). While the flocculation mechanism reported relied on electrostatic interactions between chitosan and the negative cell surface, no molecular evidence has yet confirmed this mechanism. In this study, we performed force spectroscopy AFM experiments to probe the interactions between C. vulgaris cells and chitosan at the molecular scale to decipher its flocculation mechanism. Our results showed that at pH 6, chitosan interacts with C. vulgaris cell wall through biological interactions, rather than electrostatic interactions. These observations were confirmed by comparing the data with cationically modified cellulose nanocrystals, for which the flocculation mechanism, relying on an electrostatic patch mechanism, has already been described for C. vulgaris. Further AFM experiments also showed that a different mechanism was at play at higher pH, based on chitosan precipitation. Thus this AFM-based approach highlights the complexity of chitosan-induced flocculation mechanisms for C. vulgaris.
In microalgae harvesting, flocculation is usually a compulsory preliminary step to further separation by sedimentation or flotation. For some microalgae species, and under certain growth conditions, flocculation can occur naturally. Natural flocculation presents many advantages as it does not require the addition of any flocculants to the culture medium and shows high efficiency rate. But because natural flocculation is so specific to the species and conditions, and thanks to the knowledge accumulated over the last years on flocculation mechanisms, researchers have developed strategies to induce this natural harvesting. In this review, we first decipher at the molecular scale the underlying mechanisms of natural flocculation and illustrate them by selected studies from the literature. Then we describe the developed strategies to induce natural flocculation that include the use of biopolymers, chemically modified or not, or involve mixed species cultures. But all these strategies need the addition of external compounds or microorganism which can present some issues. Thus alternative directions to completely eliminate the need for an external molecule, through genetic engineering of microalgae strains, are presented and discussed in the third part of this review.
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