Progress towards a targeted biorefinery of Chromochloris zofingiensis: a review.

Biorefinery approaches offer the potential to improve the economics of the microalgae industry by producing multiple products from a single source of biomass. Chromochloris zofingiensis shows great promise for biorefinery due to high biomass productivity and a diverse range of products including secondary carotenoids, predominantly astaxanthin; lipids such as TAGs; carbohydrates including starch; and proteins and essential amino acids. Whilst this species has been demonstrated to accumulate multiple products, the development of an integrated downstream process to obtain these is lacking. The objective of this review paper is to assess the research that has taken place and to identify the steps that must be taken to establish a biorefinery approach for C. zofingiensis. In particular, the reasons why C. zofingiensis is a promising species to target for biorefinery are discussed in terms of cellular structure, potential products, and means to accumulate desirable components via the alteration of culture conditions. Future advances and the challenges that lie ahead for successful biorefinery of this species are also reviewed along with potential solutions to address them. Supplementary Information: The online version contains supplementary material available at 10.1007/s13399-022-02955-7.

Filterability of exopolysaccharides solutions from the red microalga Porphyridium cruentum by tangential filtration on a polymeric membrane.

The red microalga Porphyridium cruentum is exploited industrially for its exopolysaccharides (EPS) and pigments production. EPS produced by P. cruentum are partially released and dissolved into the surrounding environment, they can be recovered from the culture medium after removing the cells. This paper presents a parametric study of the ultrafiltration of EPS solutions on organic membrane. The EPS solutions were produced in conditions representative of an industrial production. They were filtered at lab-scale on a flat, PES 50 kDa MWCO membrane in a complete recirculation mode of permeate and retentate. Permeate flux-transmembrane pressure (TMP) curves were established up to the limiting flux for the filtration of solutions with various values of concentration in EPS (0.10-1.06 kg GlcEq m-3), fluid tangential velocity (0.3-1.2 m s-1) and temperature (20°C and 40°C). The reversible and irreversible parts of fouling were evaluated for each experiment and the critical flux was determined for an intermediate EPS concentration (0.16 kg GlcEq m-3). The results showed that EPS solutions had a strong fouling capacity. When filtering the lowest concentrated solution (0.10 kg GlcEq m-3) with moderate fouling conditions, the overall fouling resistance was approximately half of the membrane and the share of irreversible/reversible fouling was 88% and 12%. However, the part of reversible fouling becomes predominant when approaching the limiting flux. Permeate fluxes which were obtained allow to estimate that a VRR of approximately 10 could be obtained when concentrating EPS solutions using PES membranes in flat or tubular modules but not in spiral-wound.

What Is in Store for EPS Microalgae in the Next Decade?

Microalgae and their metabolites have been an El Dorado since the turn of the 21st century. Many scientific works and industrial exploitations have thus been set up. These developments have often highlighted the need to intensify the processes for biomass production in photo-autotrophy and exploit all the microalgae value including ExoPolySaccharides (EPS). Indeed, the bottlenecks limiting the development of low value products from microalgae are not only linked to biology but also to biological engineering problems including harvesting, recycling of culture media, photoproduction, and biorefinery. Even respecting the so-called "Biorefinery Concept", few applications had a chance to emerge and survive on the market. Thus, exploiting EPS from microalgae for industrial applications in some low-value markets such as food is probably not a mature proposition considering the competitiveness of polysaccharides from terrestrial plants, macroalgae, and bacteria. However, it does not imply drawing a line on their uses but rather "thinking them" differently. This review provides insights into microalgae, EPS, and their exploitation. Perspectives on issues affecting the future of EPS microalgae are also addressed with a critical point of view.

Unravelling the matrix effect of fresh sampled cells for in vivo unbiased FTIR determination of the absolute concentration of total lipid content of microalgae.

Over the past years, the substitution of the classical biochemical quantification techniques by Fourier transform infrared (FTIR) spectroscopy has been widely studied on microalgae because of its tremendous application potential for bioprocess monitoring. In the present work, mandatory aspects that have never been approached by FTIR end-users working onto fresh biomass were assessed. We demonstrated first that fresh cells' FTIR spectra main characteristics could be severely and unspecifically altered when the properties of the sampled biomass were not monitored. Microscopy indicated that important cell reorganization could occur when diminishing the cells density of the sample. Molecular probing approach suggested that such a modification could provoke an alteration of the hydrogen-bonding network of the sample. The sample heterogeneity was found to impact also the shape and intensity of the recorded FTIR bands, participating then to a matrix effect uncharacterized until now. In the second part of our study, we selected FTIR spectra not influenced by this matrix effect and the corresponding accurate calibration data obtained by the whole cell analytical procedure to elaborate an optimized total lipid quantification PLS-R model. Results demonstrated that our strategy could provide a small volume sampling (1 mL of fresh culture), rapid (within minutes), robust (physiological condition independent), and accurate (as accurate as the reference method could be) FTIR absolute quantification method to determine the fresh microalgae intracellular total lipid content. To validate our unbiased FTIR approach, a photobioprocess monitoring pipeline was developed and allowed assessing the effect of light attenuation on total lipid production by the marine microalga Nannochloropsis oculata.

Physicochemical factors affecting the stability of two pigments: R-phycoerythrin of Grateloupia turuturu and B-phycoerythrin of Porphyridium cruentum.

Phycoerythrin is a major light-harvesting pigment of red algae, which could be used as a natural dye in foods. The stability of R-phycoerythrin of Grateloupia turuturu and B-phycoerythrin of Porphyridium cruentum in relation to different light exposure times, pHs, and temperatures was studied. Regarding the light exposure time, after 48h, the reduction in concentrations of B-phycoerythrin and R-phycoerythrin were 30±2.4% and 70±1%, respectively. Phycoerythrins presented good stability from pH 4 to 10. At pH 2, the reduction in concentration was 90±4% for B-phycoerythrin and 40±2.5% for R-phycoerythrin while, at pH 12, the phycoerythrins were degraded. Phycoerythrins showed good stability toward temperature, up to 40°C. At 60°C, the reduction in concentrations of B-phycoerythrin and R-phycoerythrin were 50±3.4% and 70±0.18%, respectively. Moreover, the best conditions of storage (-20°C) were determined.

Highly valuable microalgae: biochemical and topological aspects.

The past decade has seen a surge in the interest in microalgae culture for biodiesel production and other applications as renewable biofuels as an alternative to petroleum transport fuels. The development of new technologies for the culture of these photosynthetic microorganisms and improved knowledge of their biochemical composition has spurred innovation in the field of high-value biomolecules. These developments are only economically viable if all the microalgae fractions are valorized in a biorefinery strategy. Achieving this objective requires an understanding of microalgae content and the cellular localization of the main biomolecular families in order to develop efficient harvest and sequential recovery technologies. This review summarizes the state of the art in microalgae compositions and topologies using some examples of the main industrially farmed microalgae.

Separation and fractionation of exopolysaccharides from Porphyridium cruentum.

In this work the extraction of EPSs from culture media of Porphyridium cruentum, by dialysis, solvent-precipitation with 3 polar alcohols (methanol, ethanol and isopropanol) and membrane separation techniques has been studied. Diafiltration (DF) using a membrane with a 300 kDa molecular weight cut off was the most efficient technique compared to solvent-extraction and dialysis methods. After extraction, EPS fraction was characterized in terms of rheological properties and biochemical content. The product exhibited shear thinning behavior and a critical overlap concentration equal to 0.6 g/L. The monosaccharide composition was investigated after acidic hydrolysis. Xylose, galactose, glucose and glucuronic acid were identified as the main constitutive monomers.