Alternative Solvents for the Biorefinery of Spirulina: Impact of Pretreatment on Free Fatty Acids with High Added Value.

The growing demand for molecules of interest from microalgal biomass, such as phycobiliproteins, has led to an accumulation of unused by-products. For example, phycocyanin, obtained by the extraction of Spirulina, generated cakes rich in non-polar molecules of interest, such as free fatty acids (FFAs). These FFAs were generally considered as markers of lipidome degradation, but represented a relevant alternative to topical antibiotics, based on a biomimetic approach. In order to develop a sustainable Spirulina biorefinery scheme, different pretreatments and alternative solvents were screened to identify the best combination for the valorization of FFAs. Thus, five pre-treatments were studied including a phycocyanin extraction by-product. The following three biobased solvents were selected: ethyl acetate (EtOAc), dimethyl carbonate (DMC) and a fatty acid-based natural deep eutectic solvent (NaDES). The pigment and fatty acid profiles were established by spectroscopic and chromatographic approaches. NaDES demonstrated superior extraction capacity and selectivity compared to other biobased solvents, regardless of pretreatment. In contrast, EtOAc and DMC showed a greater diversity of FFAs, with a predominance of polyunsaturated fatty acids (PUFAs). The by-product has also been highlighted as a relevant raw material facilitating the recovery of FFAs. These results pave the way for a green biorefinery of the lipid fraction and phycobiliproteins of microalgae.

Monitoring the water content in NADES extracts from spirulina biomass by means of ATR-IR spectroscopy.

Attenuated total reflectance-infrared spectroscopy (ATR-IR) coupled with partial least squares regression (PLSR) was evaluated as a rapid, label free and cost-effective tool to quantify water content in extracts obtained from spirulina wet biomass using a glucose glycerol natural deep eutectic solvent (NADES). NADESs are green, renewable and biodegradable solvents with unique properties outcompeting existing organic solvents, for instance, for plant or biomass extraction. The properties of NADESs depend critically on their water concentration, and therefore, it is essential to develop methods to monitor it, to ensure optimal extraction efficiency and experimental repeatability to achieve a better standardization of extraction protocols. First, Karl Fischer titration was performed on a set of 20 NADES extracts in order to obtain reference water concentrations. Secondly, ATR-IR spectra were collected and subjected to datamining to construct PLSR predictive models. An R2 value of 0.9996, a mean root mean square error of cross validation of 0.136% w/w and a root mean square error of prediction of 0.130% w/w highlight the feasibility and reliability to perform quantitative analysis using ATR-IR. Moreover, the mean relative error percentage achieved, ∼0.5%, confirms the high accuracy of water concentration determination in NADES extracts. This work demonstrates that powerful alternatives are available to provide more environmentally responsible analytical protocols. ATR-IR spectroscopy applied to NADES extracts does not require any sample preparation, reagents or solvents and has minimal requirements for single use consumables. The technique is consistent with current concerns to develop greener chemistry, especially in the field of extraction of natural compounds from plants which currently represents a major focus of interest in both research and industry.

Nanovectorized Microalgal Extracts to Fight Candida albicans and Cutibacterium acnes Biofilms: Impact of Dual-Species Conditions.

Biofilm-related infections are a matter of concern especially because of the poor susceptibility of microorganisms to conventional antimicrobial agents. Innovative approaches are needed. The antibiofilm activity of extracts of cyanobacteria Arthrospira platensis, rich in free fatty acids, as well as of extract-loaded copper alginate-based nanocarriers, were studied on single- and dual-species biofilms of Candida albicans and Cutibacterium acnes. Their ability to inhibit the biofilm formation and to eradicate 24 h old biofilms was investigated. Concentrations of each species were evaluated using flow cytometry. Extracts prevented the growth of C. acnes single-species biofilms (inhibition > 75% at 0.2 mg/mL) but failed to inhibit preformed biofilms. Nanovectorised extracts reduced the growth of single-species C. albicans biofilms (inhibition > 43% at 0.2 mg/mL) while free extracts were weakly or not active. Nanovectorised extracts also inhibited preformed C. albicans biofilms by 55% to 77%, whereas the corresponding free extracts were not active. In conclusion, even if the studied nanocarrier systems displayed promising activity, especially against C. albicans, their efficacy against dual-species biofilms was limited. This study highlighted that working in such polymicrobial conditions can give a more objective view of the relevance of antibiofilm strategies by taking into account interspecies interactions that can offer additional protection to microbes.