Multi-Product Microalgae Biorefineries: From Concept Towards Reality.

Although microalgae are a promising biobased feedstock, industrial scale production is still far off. To enhance the economic viability of large-scale microalgae processes, all biomass components need to be valorized, requiring a multi-product biorefinery. However, this concept is still too expensive. Typically, downstream processing of industrial biotechnological bulk products accounts for 20-40% of the total production costs, while for a microalgae multi-product biorefinery the costs are substantially higher (50-60%). These costs are high due to the lack of appropriate and mild technologies to access the different product fractions such as proteins, carbohydrates, and lipids. To reduce the costs, simplified processes need to be developed for the main unit operations including harvesting, cell disruption, extraction, and possibly fractionation.

Understanding the salinity effect on cationic polymers in inducing flocculation of the microalga Neochloris oleoabundans.

A mechanistic study was performed to evaluate the effect of salinity on cationic polymeric flocculants, that are used for the harvesting of microalgae. The polyacrylamide Synthofloc 5080H and the polysaccharide Chitosan were employed for the flocculation of Neochloris oleoabundans. In seawater conditions, a maximum biomass recovery of 66% was obtained with a dosage of 90mg/L Chitosan. This recovery was approximately 25% lower compared to Synthofloc 5080H reaching recoveries greater than 90% with dosages of 30mg/L. Although different recoveries were obtained with both flocculants, the polymers exhibit a similar apparent polymer length, as was evaluated from viscosity measurements. While both flocculants exhibit similar polymer lengths in increasing salinity, the zeta potential differs. This indicates that polymeric charge dominates flocculation. With increased salinity, the effectivity of cationic polymeric flocculants decreases due to a reduction in cationic charge. This mechanism was confirmed through a SEM analysis and additional experiments using flocculants with various charge densities.

Dosage effect of cationic polymers on the flocculation efficiency of the marine microalga Neochloris oleoabundans.

A mechanistic mathematical model was developed to predict the performance of cationic polymers for flocculating salt water cultivated microalgae. The model was validated on experiments carried out with Neochloris oleoabundans and three different commercial flocculants (Zetag 7557®, Synthofloc 5080H® and SNF H536®). For a wide range of biomass concentrations (0.49-1.37 g L(-1)) and flocculant dosages (0-150 mg L(-1)) the model simulations predicted well the optimal flocculant-to-biomass ratio between 43 and 109 mgflocculant/gbiomass. At optimum conditions biomass recoveries varied between 88% and 99%. The cost of the usage of commercial available flocculants is estimated to range between 0.15$/kgbiomass and 0.49$/kgbiomass.

Cationic polymers for successful flocculation of marine microalgae.

Flocculation of microalgae is a promising technique to reduce the costs and energy required for harvesting microalgae. Harvesting marine microalgae requires suitable flocculants to induce the flocculation under marine conditions. This study demonstrates that cationic polymeric flocculants can be used to harvest marine microalgae. Different organic flocculants were tested to flocculate Phaeodactylum tricornutum and Neochloris oleoabundans grown under marine conditions. Addition of 10 ppm of the commercial available flocculants Zetag 7557 and Synthofloc 5080H to P. tricornutum showed a recovery of, respectively, 98% ± 2.0 and 94% ± 2.9 after flocculation followed by 2h sedimentation. Using the same flocculants and dosage for harvesting N. oleoabundans resulted in a recovery of 52% ± 1.5 and 36% ± 11.3. This study shows that cationic polymeric flocculants are a viable option to pre-concentrate marine cultivated microalgae via flocculation prior to further dewatering.

Mechanism behind autoflocculation of unicellular green microalgae Ettlia texensis.

The oleaginous Ettlia texensis is an autoflocculating green microalga that can be used for bio-flocculation of other microalgae species to facilitate harvesting. In this study the mechanism behind autoflocculation of E. texensis was revealed by scanning electron microscopy (SEM) analysis and by characterisation of the cell surface properties. SEM analysis and measurement of extracellular polymeric substances (EPS) showed that autoflocculation of E. texensis is due to the EPS containing mainly glycoproteins patched to the cell surface. Despite the presence of charged groups on the cell surface, they do not seem to attribute to autoflocculation of E. texensis. During bio-flocculation of E. texensis with Chlorella vulgaris EPS structures between both microalgal species were observed. EPS thus not only play a predominant role in autoflocculation of E. texensis but also in bio-flocculation when using this microalga to harvest others.

Modeling microalgal flocculation and sedimentation.

In this study, a combined flocculation and sedimentation model is developed. The model predicts the time needed to reach a desired concentration of microalgal suspension in a sedimentation tank. The concentration of the particles as function of the time and the position in the tank is described. The model was validated with experimental data for Ettlia texensis. The concentration changes measured in time at different heights in the sedimentation vessel corresponded well with model predictions. The model predicts that it takes 25 h to reach a final concentration of 5.2 gDW L(-1), when the initial concentration is 0.26 gDW L(-1) and the tank height is 1m. This example illustrates the use of this model for the design of the settling tank needed for pre-concentration of microalgal biomass before further dewatering.

Effect of growth phase on harvesting characteristics, autoflocculation and lipid content of Ettlia texensis for microalgal biodiesel production.

The effect of growth phase on the recovery of the autoflocculating microalgae Ettlia texensis was studied. In the stationary phase, 90% recovery was achieved after 3h settling. Scanning electron microscopic pictures revealed that extracellular polymeric substances (EPS) on the cell surface were involved in autoflocculation. During the stationary phase an increase of the protein fraction in the EPS was observed while the total fatty acids content increased. The autoflocculating properties of E. texensis combined with favourite fatty acid content and composition make this microalgae an excellent candidate for biodiesel production if harvested at the end of the stationary phase.

Ratio between autoflocculating and target microalgae affects the energy-efficient harvesting by bio-flocculation.

The effect of ratio between autoflocculating and target microalgae in bio-flocculation was studied with emphasis on the recovery, sedimentation rate and energy demand for harvesting the target microalgae. When the autoflocculating microalgae Ettlia texensis, Ankistrodesmus falcatus and Scenedesmus obliquus were added to Chlorella vulgaris at a ratio of 0.25, the recovery of C. vulgaris increased from 25% to, respectively, 40%, 36% and 31%. The sedimentation rate increased as well. Addition of Tetraselmis suecica to Neochloris oleoabundans at a ratio of 0.25 increased the recovery from 40% to 50%. Application of bio-flocculation at a ratio of 0.25, followed by centrifugation reduces the energy demand for harvesting of the target microalgae from 13.8 MJ kgDW(-1) if only centrifugation is used to 1.83, 1.81, 1.53 and 1.34 MJ kgDW(-1), respectively, using T. suecica, E. texensis, A. falcatus and S. obliquus and 3h sedimentation before centrifugation.

Growth inhibition of Monodus subterraneus by free fatty acids.

Monodus subterraneus is a microalga, which is known for its high eicosapentaenoic acid (EPA; 20:5omega3) content. To produce EPA commercially, high volumetric productivities of microalgae are required. These high productivities can be reached in flat panel photobioreactors with small optical paths that have to be operated at high cell densities (>10 g/L). However, at these cell densities a reduction of productivity is observed. This growth inhibition is probably caused by growth inhibitors released by the microalgae, which have been suggested to be fatty acids. Our aim was to investigate if free fatty acids produced by M. subterraneus inhibited growth of this species. Therefore a bioassay was developed and saturated, unsaturated and poly-unsaturated fatty acids occurring in Monodus were tested on their growth inhibiting properties. Growth of M. subterraneus was completely inhibited at a saturated concentration (96 microM) of palmitoleic acid (16:1omega7). But, the saturated fatty acid palmitic acid (16:0) and the mono-saturated oleic acid (18:1omega9) were much stronger inhibitors. Growth was inhibited for 50% already at concentrations of 0.4 microM 16:0 and 3 microM 18:1omega9, respectively. These fatty acids probably cause the growth inhibition in high cell density cultures of M. subterraneus.