Cultivation and Biorefinery of Microalgae (Chlorella sp.) for Producing Biofuels and Other Byproducts: A Review

Microalgae-based carbon dioxide (CO2) biofixation and biorefinery are the most efficient methods of biological CO2 reduction and reutilization. The diversification and high-value byproducts of microalgal biomass, known as microalgae-based biorefinery, are considered the most promising platforms for the sustainable development of energy and the environment, in addition to the improvement and integration of microalgal cultivation, scale-up, harvest, and extraction technologies. In this review, the factors influencing CO2 biofixation by microalgae, including microalgal strains, flue gas, wastewater, light, pH, temperature, and microalgae cultivation systems are summarized. Moreover, the biorefinery of Chlorella biomass for producing biofuels and its byproducts, such as fine chemicals, feed additives, and high-value products, are also discussed. The technical and economic assessments (TEAs) and life cycle assessments (LCAs) are introduced to evaluate the sustainability of microalgae CO2 fixation technology. This review provides detailed insights on the adjusted factors of microalgal cultivation to establish sustainable biological CO2 fixation technology, and the diversified applications of microalgal biomass in biorefinery. The economic and environmental sustainability, and the limitations and needs of microalgal CO2 fixation, are discussed. Finally, future research directions are provided for CO2 reduction by microalgae.

Assuaging Microalgal Harvesting Woes via Attached Growth: A Critical Review to Produce Sustainable Microalgal Feedstock

Third-generation biofuels that are derived from microalgal biomass have gained momentum as a way forward in the sustainable production of biodiesel. Such efforts are propelled by the intention to reduce our dependence on fossil fuels as the primary source of energy. Accordingly, growing microalgal biomass in the form of suspended cultivation has been a conventional technique for the past few decades. To overcome the inevitable harvesting shortcomings arising from the excessive energy and time needed to separate the planktonic microalgal cells from water medium, researchers have started to explore attached microalgal cultivation systems. This cultivation mode permits the ease of harvesting mature microalgal biomass, circumventing the need to employ complex harvesting techniques to single out the cells, and is economically attractive. However, the main bottleneck associated with attached microalgal growth is low biomass production due to the difficulties the microalgal cells have in forming attachment and populating thereafter. In this regard, the current review encompasses the novel techniques adopted to promote attached microalgal growth. The physicochemical effects such as the pH of the culture medium, hydrophobicity, as well as the substratum surface properties and abiotic factors that can determine the fate of exponential growth of attached microalgal cells, are critically reviewed. This review aims to unveil the benefits of an attached microalgal cultivation system as a promising harvesting technique to produce sustainable biodiesel for lasting applications.

Biodegradable, metal-chelating compounds as alternatives to EDTA for cultivation of marine microalgae

Iron (Fe) is an essential nutrient for microalgal metabolism. The low solubility of Fe in oxic aquatic environments can be a growth-limiting factor for phytoplankton. Synthetic chelating agents, such as ethylenediaminetetraacetic acid (EDTA), are used widely to maintain Fe in solution for microalgal cultivation. The non-biodegradable nature of EDTA, combined with sub-optimal bioavailability of Fe-EDTA complexes to microalgae, indicates opportunity to improve microalgal cultivation practices that amplify production efficiency and environmental compatibility. In the present study, the effects of four organic chelating ligands known to form readily bioavailable organic complexes with Fe in natural aquatic environments were investigated in relation to growth and biochemical composition of two marine microalgae grown as live feeds in shellfish hatcheries (Chaetoceros calcitrans and Tisochrysis lutea). Three saccharides, alginic acid (ALG), glucuronic acid (GLU), and dextran (DEX), as well as the siderophore desferrioxamine B (DFB), were compared to EDTA. Organic ligands characterized by weaker binding capacity for cationic metals (i.e., ALG, GLU, DEX) significantly improved microalgal growth and yields in laboratory-scale static batch cultures or bubbled photobioreactors. Maximal microalgal growth enhancement relative to the control (e.g., EDTA) was recorded for GLU, followed by ALG, with 20–35% increase in specific growth rate in the early stages of culture development of C. calcitrans and T. lutea. Substitution of EDTA with GLU resulted in a 27% increase in cellular omega 3-polyunsaturetd fatty acid content of C. calcitrans and doubled final cell yields. Enhanced microalgal culture performance is likely associated with increased intracellular Fe uptake efficiency combined with heterotrophic growth stimulated by the organic ligands. Based upon these results, we propose that replacement of EDTA with one of these organic metal-chelating ligands is an effective and easily implementable strategy to enhance the environmental compatibility of microalgal cultivation practices while also maximizing algal growth and enhancing the nutritional quality of marine microalgal species commonly cultured for live-feed applications in aquaculture.

Phosphite Reduces the Predation Impact of Poterioochromonas malhamensis on Cyanobacterial Culture

Contamination by the predatory zooplankton Poterioochromonas malhamensis is one of the major threats that causes catastrophic damage to commercial-scale microalgal cultivation. However, knowledge of how to manage predator contamination is limited. Previously, we established a phosphite (Pt)-based culture system by engineering Synechococcus elongatus, which exerted a competitive growth advantage against microbial contaminants that compete with phosphate source. Here, we examined whether Pt is effective in suppressing predator-type contamination. Co-culture experiment of Synechococcus with isolated P. malhamensis revealed that, although an addition of Pt at low concentrations up to 2.0 mM was not effective, increased dosage of Pt (~20 mM) resulted in the reduced grazing impact of P. malhamensis. By using unsterilized raw environmental water collected from rivers or ponds, we found that the suppression effect of Pt was dependent on the type of environmental water used. Eukaryotic microbial community analysis of the cultures using environmental water samples revealed that Paraphysomonas, a colorless Chrysophyceae, emerged and dominated under high-Pt conditions, suggesting that Paraphysomonas is insensitive to Pt compared to P. malhamensis. These findings may provide a clue for developing a strategy to reduce the impact of grazer contamination in commercial-scale microalgal cultivation.

Functional Diversity Facilitates Stability Under Environmental Changes in an Outdoor Microalgal Cultivation System

Functionally uniform monocultures have remained the paradigm in microalgal cultivation despite the apparent challenges to avoid invasions by other microorganisms. A mixed microbial consortium approach has the potential to optimize and maintain biomass production despite of seasonal changes and to be more resilient toward contaminations. Here we present a 3-year outdoor production of mixed consortia of locally adapted microalgae and bacteria in cold temperate latitude. Microalgal consortia were cultivated in flat panel photobioreactors using brackish Baltic Sea water and CO2 from a cement factory (Degerhamn, Cementa AB, Heidelberg Cement Group) as a sustainable CO2 source. To evaluate the ability of the microbial consortia to maintain stable biomass production while exposed to seasonal changes in both light and temperature, we tracked changes in the microbial community using molecular methods (16S and 18S rDNA amplicon sequencing) and monitored the biomass production and quality (lipid, protein, and carbohydrate content) over 3 years. Despite changes in environmental conditions, the mixed consortia maintained stable biomass production by alternating between two different predominant green microalgae (Monoraphidium and Mychonastes) with complementary tolerance to temperature. The bacterial population was few taxa co-occured over time and the composition did not have any connection to the shifts in microalgal taxa. We propose that a locally adapted and mixed microalgal consortia, with complementary traits, can be useful for optimizing yield of commercial scale microalgal cultivation.