Microalgae-mediated biofixation as an innovative technology for flue gases towards carbon neutrality: A comprehensive review.

Microalgae-mediated industrial flue gas biofixation has been widely discussed as a clean alternative for greenhouse gas mitigation. Through photosynthetic processes, microalgae can fix carbon dioxide (CO2) and other compounds and can also be exploited to obtain high value-added products in a circular economy. One of the major limitations of this bioprocess is the high concentrations of CO2, sulfur oxides (SOx), and nitrogen oxides (NOx) in flue gases, according to the origin of the fuel, that can inhibit photosynthesis and reduce the process efficiency. To overcome these limitations, researchers have recently developed new technologies and enhanced process configurations, thereby increased productivity and CO2 removal rates. Overall, CO2 biofixation rates from flue gases by microalgae ranged from 72 mg L-1 d -1 to over 435 mg L-1 d-1, which were directly influenced by different factors, mainly the microalgae species and photobioreactor. Additionally, mixotrophic culture have shown potential in improving microalgae productivity. Progress in developing new reactor configurations, with pilot-scale implementations was observed, resulting in an increase in patents related to the subject and in the implementation of companies using combustion gases in microalgae culture. Advancements in microalgae-based green technologies for environmental impact mitigation have led to more efficient biotechnological processes and opened large-scale possibilities.

Enhancing environmental performance in biogas production from wastewater-grown microalgae: A life cycle assessment perspective.

The production of biogas from microalgae has gained attention due to their rapid growth, CO2 sequestration, and minimal land use. This study uses life cycle assessment to assess the environmental impacts of biogas production from wastewater-grown microalgae through anaerobic digestion within an optimized microalgae-based system. Using SimaPro® 9 software, 3 scenarios were modeled considering the ReCiPe v1.13 midpoint and endpoint methods for environmental impact assessment in different categories. In the baseline scenario (S1), a hypothetical system for biogas production was considered, consisting of a high rate algal pond (HRAP), a settling, an anaerobic digester, and a biogas upgrading unit. The second scenario (S2) included strategies to enhance biogas yield, namely co-digestion and thermal pre-treatment. The third scenario (S3), besides considering the strategies of S2, proposed the biogas upgrading in the HRAP and the digestate recovery as a biofertilizer. After normalization, human carcinogenic toxicity was the most positively affected category due to water use in the cultivation step, accounted as avoided product. However, this category was also the most negatively affected by the impacts of the digester heating energy. Anaerobic digestion was the most impactful step, constituting on average 60.37% of total impacts. Scenario S3 performed better environmentally, primarily due to the integration of biogas upgrading within the cultivation reactor and digestate use as a biofertilizer. Sensitivity analysis highlighted methane yield's importance, showing potential for an 11.28% reduction in ionizing radiation impacts with a 10% increase. Comparing S3 biogas with natural gas, the resource scarcity impact was reduced sixfold, but the human health impact was 23 times higher in S3.

Effect of microalgae and bacteria inoculation on the startup of bioreactors for paper pulp wastewater and biofuel production.

The use of microalgae and bacteria as a strategy for the startup of bioreactors for the treatment of industrial wastewater can be a sustainable and economically viable alternative. This technology model provides satisfactory results in the nitrification and denitrification process for nitrogen removal, organic matter removal, biomass growth, sedimentation, and byproducts recovery for added-value product production. The objective of this work was to evaluate the performance of microalgae and bacteria in their symbiotic process when used in the treatment of paper pulp industry wastewater. The experiment, lasting fourteen days, utilized four bioreactors with varying concentrations in mgVSS/L of microalgae to bacteria ratio (R1-100:100, R2-100:300, R3-100:500, R4-300:100) in the startup process. Regarding the sludge volumetric index (SVI), the results show that the R1 and R2 reactors developed SVI30/SVI10 biomass in the range of 85.57 ± 7.33% and 84.72 ± 8.19%, respectively. The lipid content in the biomass of reactors R1, R2, R3 e R4 was 13%, 7%, 19%, and 22%, respectively. This high oil content at the end of the batch, may be related to the nutritional stress that the species underwent during this feeding regime. In terms of chlorophyll, the bioreactor with an initial inoculation of 100:100 showed better symbiotic growth of microalgae and bacteria, allowing exponential growth of microalgae. The total chlorophyll value for this bioreactor was 801.46 ± 196.96 µg/L. Biological removal of nitrogen from wastewater from the paper pulp industry is a challenge due to the characteristics of the effluent, but the four reactors operated in a single batch obtained good nitrogen removal. Ammonia nitrogen removal performances were 91.55 ± 9.99%, 72.13 ± 19.18%, 64.04 ± 21.34%, and 86.15 ± 30.10% in R1, R2, R3, and R4, respectively.

Cultivation of yeasts on liquid digestate to remove organic pollutants and nutrients and for potential application as co-culture with microalgae.

In this study, the growth of yeast and yeast-like fungi in the liquid digestate from vegetable wastes was investigated in order to remove nutrients and organic pollutants, and for their application as co-culture members with green microalgae. The studied yeast strains were characterized for their assimilative and enzymatic profiles as well as temperature requirements. In the first experimental stage, the growth dynamics of each strain were determined, allowing to select the best yeasts for further studies. In the subsequent stage, the ability of selectants to remove organic pollutants was assessed. Different cultivation media containing respectively 1:3, 1:1, 3:1 vol ratio of liquid digestate and the basal minimal medium were used. Among all tested yeast strains, Rhodotorula mucilaginosa DSM 70825 showed the most promising results, demonstrating the highest potential for removing organic substrates and nutrients. Depending on the medium, this strain achieved 50-80% sCOD, 45-60% tVFAs, 21-45% TN, 33-52% PO43- reduction rates. Similar results were obtained for the strain Candida sp. OR687571. The high nutrient and organics removal efficiency by these yeasts could likely be linked to their ability to assimilate xylose (being the main source of carbon in the liquid digestate). In culture media containing liquid digestate, both yeast strains achieved good viability and proliferation potential. In the liquid digestate medium, R. mucilaginosa and Candida sp. showed vitality at the level of 51.5% and 45.0%, respectively. These strains seem to be a good starting material for developing effective digestate treatment strategies involving monocultures and/or consortia with other yeasts or green microalgae.

Factors impacting the microbial production of eicosapentaenoic acid.

The increasing applications for eicosapentaenoic acid (EPA) and the potential shortfall in supply due to sustainability and contamination issues related with its conventional sources (i.e., fish oils; seafood) led to an extensive search for alternative and sustainable sources, as well as production processes. The present mini-review covers all the steps involved in the production of EPA from microorganisms, with a deeper focus on microalgae. From production systems to downstream processing, the most important achievements within each area are briefly highlighted. Comparative tables of methodologies are also provided, as well as additional references of recent reviews, so that readers may deepen their knowledge in the different issues addressed. KEY POINTS: • Microorganisms are more sustainable alternative sources of EPA than fish. • Due to the costly separation from DHA, species that produce only EPA are preferable. • EPA production can be optimised using non-genetic and genetic tailoring engineering.

Bioremediation potential of microalgae for sustainable soil treatment in India: A comprehensive review on heavy metal and pesticide contaminant removal.

The escalating environmental concerns arising from soils contamination with heavy metals (HMs) and pesticides (PSTs) necessitate the development of sustainable and effective remediation strategies. These contaminants, known for their carcinogenic properties and toxicity even at small amounts, pose significant threats to both environmental ecology and human health. While various chemical and physical treatments are employed globally, their acceptance is often hindered by prolonged remediation times, high costs, and inefficacy in areas with exceptionally high pollutant concentrations. A promising emerging trend in addressing this issue is the utilization of microalgae for bioremediation. Bioremediation, particularly through microalgae, presents numerous benefits such as high efficiency, low cost, easy accessibility and an eco-friendly nature. This approach has gained widespread use in remediating HM and PST pollution, especially in large areas. This comprehensive review systematically explores the bioremediation potential of microalgae, shedding light on their application in mitigating soil pollutants. The paper summarizes the mechanisms by which microalgae remediate HMs and PSTs and considers various factors influencing the process, such as pH, temperature, pollutant concentration, co-existing pollutants, time of exposure, nutrient availability, and light intensity. Additionally, the review delves into the response and tolerance of various microalgae strains to these contaminants, along with their bioaccumulation capabilities. Challenges and future prospects in the microalgal bioremediation of pollutants are also discussed. Overall, the aim is to offer valuable insights to facilitate the future development of commercially viable and efficient microalgae-based solutions for pollutant bioremediation.

Characterization and potentiality of plant-derived silver nanoparticles for enhancement of biomass and hydrogen production in Chlorella sp. under nitrogen deprived condition.

Energy is a crucial entity for the development and it has various alternative forms of energy sources. Recently, the synthesis of nanoparticles using benign biocatalyst has attracted increased attention. In this study, silver nanoparticles were synthesized and characterized using Azadirachta indica plant-derived phytochemical as the reducing agent. Biomass of the microalga Chlorella sp. cultivated in BG11 medium increased after exposure to low concentrations of up to 0.48 mg L-1 AgNPs. In addition, algal cells treated with 0.24 mg L-1 AgNPs and cultivated in BG110 medium which contained no nitrogen source showed the highest hydrogen yield of 10.8 mmol L-1, whereas the untreated cells under the same conditions showed very low hydrogen yield of 0.003 mmol L-1. The enhanced hydrogen production observed in the treated cells was consistent with an increase in hydrogenase activity. Treatment of BG110 grown cells with low concentration of green synthesized AgNPs at 0.24 mg L-1 enhanced hydrogenase activity with a 5-fold increase of enzyme activity compared to untreated BG110 grown cells. In addition, to improve photolytic water splitting efficiency for hydrogen production, cells treated with AgNPs at 0.24 mg L-1 showed highest oxygen evolution signifying improvement in photosynthesis. The silver nanoparticles synthesized using phytochemicals derived from plant enhanced both microalgal biomass and hydrogen production with an added advantage of CO2 reduction which could be achieved due to an increase in biomass. Hence, treating microalgae with nanoparticles provided a promising strategy to reduce the atmospheric carbon dioxide as well as increasing production of hydrogen as clean energy.

Assessing nitrous oxide emissions from algal-bacterial photobioreactors devoted to biogas upgrading and digestate treatment.

Nitrous oxide (N2O) emissions in High Rate Algal Ponds (HRAP) can negatively affect the sustainability of algal-bacterial processes. N2O emissions from a pilot HRAP devoted to biogas upgrading and digestate treatment were herein monitored for 73 days. The influence of the pH (7.5, 8.5, and 9.5), nitrogen sources (100 mg L-1 of N-NO2-, N-NO3-, and N-NH4+) and illumination on N2O emissions from the algal-bacterial biomass of the HRAP was also assessed in batch tests. Significantly higher N2O gas concentrations of 311.8 ± 101.1 ppmv were recorded in the dark compared to the illuminated period (236.9 ± 82.6 ppmv) in the HRAP. The batch tests revealed that the highest N2O emission rates (49.4 mmol g-1 TSS·h-1) occurred at pH 8.5 in the presence of 100 mg N-NO2-/L under dark conditions. This study revealed significant N2O emissions in HRAPs during darkness.

Microalgae cultivation trials in a membrane bioreactor operated in heterotrophic, mixotrophic, and phototrophic modes using ammonium-rich wastewater: The study of fouling.

In this work, microalgae cultivation trials were carried out in a membrane bioreactor to investigate fouling when the cultures of Chlorellavulgaris were grown under mixotrophic, heterotrophic, and phototrophic cultivation regimes. The Chlorella cultures were cultivated in wastewater as a source of nutrients that contained a high concentration of ammonium. In mixotrophic cultivation trials, the results showed that the elevated contents of carbohydrates in the soluble microbial product and proteins in extracellular polymeric substances probably initiated membrane fouling. In this case, the highest protein content was also found in extracellular polymeric substances due to the high nitrogen removal rate. Consequently, transmembrane pressure significantly increased compared to the phototrophic and heterotrophic regimes. The data indicated that cake resistance was the main cause of fouling in all cultivations. Higher protein content in the cake layer made the membrane surface more hydrophobic, while carbohydrates had the opposite effect. Compared to a mixotrophic culture, a phototrophic culture had a larger cell size and higher hydrophobicity, leading to less membrane fouling. Based on our previous data, the highest ammonia removal rate was reached in the mixotrophic cultures; nevertheless, membrane fouling appeared to be the fundamental problem.

Cohesive phycoremediation of pyrene by freshwater microalgae Selenastrum sp. and biodiesel production and its assessment.

In this study, the freshwater microalgae Selenastrum sp. was assessed for the effective degradation of pyrene and simultaneous production of biodiesel from pyrene-tolerant biomass. The growth of algae was determined based on the cell dry weight, cell density, chlorophyll content, and biomass productivity under different pyrene concentrations. Further, lipids from pyrene tolerant culture were converted into biodiesel by acid-catalyzed transesterification, which was characterized for the total fatty acid profile by gas chromatography. Increased pyrene concentration revealed less biomass yield and productivity after 20 days of treatment, indicating potent pyrene biodegradation by Selenastrum sp. Biomass yield was unaffected till the 20 mg/L pyrene. A 95% of pyrene bioremediation was observed at 20 days of culturing. Lipid accumulation of 22.14%, as evident from the estimation of the total lipid content, indicated a marginal increase in corroborating pyrene stress in the culture. Fatty acid methyl esters yield of 63.06% (% per 100 g lipids) was noticed from the pyrene tolerant culture. Moreover, fatty acid profile analysis of biodiesel produced under 10 mg/L and 20 mg/L pyrene condition showed escalated levels of desirable fatty acids in Selenastrum sp., compared to the control. Further, Selenastrum sp. and other freshwater microalgae are catalogued for sustainable development goals attainment by 2030, as per the UNSDG (United Nations Sustainable Development Goals) agenda. Critical applications for the Selenastrum sp. in bioremediation of pyrene, along with biodiesel production, are enumerated for sustainable and renewable energy production and resource management.

Central transcriptional regulator controls photosynthetic growth and carbon storage in response to high light.

Carbon capture and biochemical storage are some of the primary drivers of photosynthetic yield and productivity. To elucidate the mechanisms governing carbon allocation, we designed a photosynthetic light response test system for genetic and metabolic carbon assimilation tracking, using microalgae as simplified plant models. The systems biology mapping of high light-responsive photophysiology and carbon utilization dynamics between two variants of the same Picochlorum celeri species, TG1 and TG2 elucidated metabolic bottlenecks and transport rates of intermediates using instationary 13C-fluxomics. Simultaneous global gene expression dynamics showed 73% of the annotated genes responding within one hour, elucidating a singular, diel-responsive transcription factor, closely related to the CCA1/LHY clock genes in plants, with significantly altered expression in TG2. Transgenic P. celeri TG1 cells expressing the TG2 CCA1/LHY gene, showed 15% increase in growth rates and 25% increase in storage carbohydrate content, supporting a coordinating regulatory function for a single transcription factor.

The Biophysical Properties of Microalgal Cell Surfaces Govern Their Interactions with an Amphiphilic Chitosan Derivative Used for Flocculation and Flotation.

Microalgae show great promise for producing valuable molecules like biofuels, but their large-scale production faces challenges, with harvesting being particularly expensive due to their low concentration in water, necessitating extensive treatment. While methods such as centrifugation and filtration have been proposed, their efficiency and cost-effectiveness are limited. Flotation, involving air-bubbles lifting microalgae to the surface, offers a viable alternative, yet the repulsive interaction between bubbles and cells can hinder its effectiveness. Previous research from our group proposed using an amphiphilic chitosan derivative, polyoctyl chitosan (PO-chitosan), to functionalize bubbles used in dissolved air flotation (DAF). Molecular-scale studies performed using atomic force microscopy (AFM) revealed that PO-chitosan's efficiency correlates with cell surface properties, particularly hydrophobic ones, raising the question of whether this molecule can in fact be used more generally to harvest different microalgae. Evaluating this, we used a different strain of Chlorella vulgaris and first characterized its surface properties using AFM. Results showed that cells were hydrophilic but could still interact with PO-chitosan on bubble surfaces through a different mechanism based on specific interactions. Although force levels were low, flotation resulted in 84% separation, which could be explained by the presence of AOM (algal organic matter) that also interacts with functionalized bubbles, enhancing the overall separation. Finally, flocculation was also shown to be efficient and pH-independent, demonstrating the potential of PO-chitosan for harvesting microalgae with different cell surface properties and thus for further sustainable large-scale applications.

Nickel adsorbed algae biochar based oxygen reduction reaction catalyst.

Lately, the bio electrochemical systems are emerging as an efficient wastewater treatment and energy conversion technology. However, their scaling-up is considerably restrained by slow-rate of cathodic oxygen reduction reaction (ORR) or otherwise by the high cost associated with the available efficient ORR catalysts. In this investigation, a cost-effective and eco-friendly approach for synthesizing Ni based ORR catalyst utilizing biosorption property of microalgae is accomplished. The synthesised Ni adsorbed algal biochar (NAB) served as an efficient cathode catalyst for enhancing ORR in a microbial carbon-capture cell (MCC). On increasing the initial concentration of Ni2+ in the aqueous medium from 100 mgL-1 to 500 mgL-1, the biosorption capacity was found to increase from 3 mgg-1 to 32 mgg-1 of algae cell. The MCC operated with NAB based cathode catalyst loading of 2 mgcm-2 exhibited 3.5 times higher power density (4.69 Wm-3) as compared to the one with commercial activated carbon. A significant organic matter removal (82 %) in the anodic chamber with simultaneous algal biomass productivity in the cathodic chamber was attained by MCC with cathode loaded with 2 mgcm-2 of NAB. Hence, this easily synthesised low-cost catalyst, out of waste stream, proved its ability to improve the performance of MCC.

Developing biomass augmentation strategy for cultivation of Marvania coccoides using fruit waste and wastewater based growth medium for biodiesel production.

Microalgae cultivation using waste as nutrient source can minimize the use of expensive chemical nutrients and valuable freshwater. In present work, novel microalgae Marvania coccoides was cultivated in fruit waste (FW) and wastewater (WW) based growth medium. To further augment metabolites and biomass, the culture was supplemented with phytohormone, kinetin (K). Various pre-treatment methods were investigated for improving the nutrient release and bacterial load reduction in waste-based medium. Phytohormone supplemented fruit waste and wastewater media (WW + FW + K) showed improved biomass productivity of 117.14 mg.L-1.d-1 compared to both waste-based and synthetic medium. Biomass harvested from WW + FW + K showed increased lipid content (22 %) as compared to lipid content (19 %) of biomass from synthetic medium. Biodiesel yield of 91.50 % was observed with fatty acids C16:0, C16:2, C18:0, C18:1, C18:2, C19:0, C20:1, C20:2 and C22:1 in composition. M. coccoides can be cultivated in waste medium and used as promising feedstock for production of biodiesel.

A comprehensive review on the heterotrophic production of bioactive compounds by microalgae.

Bioactive compounds derived from microalgae have garnered considerable attention as valuable resources for drugs, functional foods, and cosmetics. Among these compounds, photosynthetic pigments and polyunsaturated fatty acids (PUFAs) have gained increasing interest due to their numerous beneficial properties, including anti-oxidant, anti-viral, anti-bacterial, anti-fungal, anti-inflammatory, and anti-tumor effects. Several microalgae species have been identified as rich sources of bioactive compounds, including the Chlorophyceae Dunaliella and Haematococcus, the Bacillariophyta Phaeodactylum and Nitzschia, and the dinoflagellate Crypthecodinium cohnii. However, most of the reported microalgae species primarily grow through autotrophic mechanisms, resulting in low yields and high production costs of bioactive compounds. Consequently, the utilization of heterotrophic microalgae, such as Chromochloris zofingiensis and Nitzschia laevis, has shown significant advantages in the production of astaxanthin and eicosapentaenoic acid (EPA), respectively. These heterotrophic microalgae exhibit superior capabilities in synthesizing target compounds. This comprehensive review provides a thorough examination of the heterotrophic production of bioactive compounds by microalgae. It covers key aspects, including the metabolic pathways involved, the impact of cultivation conditions, and the practical applications of these compounds. The review discusses how heterotrophic cultivation strategies can be optimized to enhance bioactive compound yields, shedding light on the potential of microalgae as a valuable resource for high-value product development.

Light management by algal aggregates in living photosynthetic hydrogels.

Rapid progress in algal biotechnology has triggered a growing interest in hydrogel-encapsulated microalgal cultivation, especially for the engineering of functional photosynthetic materials and biomass production. An overlooked characteristic of gel-encapsulated cultures is the emergence of cell aggregates, which are the result of the mechanical confinement of the cells. Such aggregates have a dramatic effect on the light management of gel-encapsulated photobioreactors and hence strongly affect the photosynthetic outcome. To evaluate such an effect, we experimentally studied the optical response of hydrogels containing algal aggregates and developed optical simulations to study the resultant light intensity profiles. The simulations are validated experimentally via transmittance measurements using an integrating sphere and aggregate volume analysis with confocal microscopy. Specifically, the heterogeneous distribution of cell aggregates in a hydrogel matrix can increase light penetration while alleviating photoinhibition more effectively than in a flat biofilm. Finally, we demonstrate that light harvesting efficiency can be further enhanced with the introduction of scattering particles within the hydrogel matrix, leading to a fourfold increase in biomass growth. Our study, therefore, highlights a strategy for the design of spatially efficient photosynthetic living materials that have important implications for the engineering of future algal cultivation systems.

Sulfadiazine proliferated antibiotic resistance genes in the phycosphere of Chlorella pyrenoidosa: Insights from bacterial communities and microalgal metabolites.

The phycosphere is an essential ecological niche for the proliferation of antibiotic resistance genes (ARGs). However, how ARGs' potential hosts change and the driving mechanism of metabolites under antibiotic stress in the phycosphere have seldom been researched. We investigated the response of Chlorella pyrenoidosa and the structure and abundance of free-living (FL) and particle-attached (PA) bacteria, ARGs, and metabolites under sulfadiazine by using real-time quantitative PCR, 16 S rRNA high-throughput. The linkage of key bacterial communities, ARGs, and metabolites through correlations was established. Through analysis of physiological indicators, Chlorella pyrenoidosa displayed a pattern of "low-dose promotion and high-dose inhibition" under antibiotic stress. ARGs were enriched in the PA treatment groups by 117 %. At the phylum level, Proteobacteria, Bacteroidetes, and Actinobacteria as potential hosts for ARGs. At the genus level, potential hosts included Sphingopyxis, SM1A02, Aquimonas, Vitellibacter, and Proteiniphilum. Middle and high antibiotic concentrations induced the secretion of metabolites closely related to potential hosts by algae, such as phytosphingosine, Lysophosphatidylcholine, and α-Linolenic acid. Therefore, changes in bacterial communities indirectly influenced the distribution of ARGs through alterations in metabolic products. These findings offer essential details about the mechanisms behind the spread and proliferation of ARGs in the phycosphere.

Investigating quantitative approach for microalgal biomass using deep convolutional neural networks and image recognition.

The effective monitoring of microalgae cultivation is crucial for optimizing their energy utilization efficiency. In this paper, a quantitative analysis method, using microalgae images based on two convolutional neural networks, EfficientNet (EFF) and residual network (RES), is proposed. Suspension samples prepared from two types of dried microalgae powders, Rhodophyta (RH) and Spirulina (SP), were used to mimic real microalgae cultivation settings. The method's prediction accuracy of the algae concentration ranges from 0.94 to 0.99. RH, with a distinctively pronounced red-green-blue value shift, achieves a higher prediction accuracy than SP. The prediction results of the two algorithms were significantly superior to those of a linear regression. Additionally, RES outperforms EFF in terms of its generalization ability and robustness, which is attributable to its distinct residual block architecture. The RES provides a viable approach for the image-based quantitative analysis.

Evolving perspectives on lutein production from microalgae - A focus on productivity and heterotrophic culture.

Increased consumer awareness for healthier and more sustainable products has driven the search for naturally sourced compounds as substitutes for chemically synthesized counterparts. Research on pigments of natural origin, such as carotenoids, particularly lutein, has been increasing for over three decades. Lutein is recognized for its antioxidant and photoprotective activity. Its ability to cross the blood-brain barrier allows it to act at the eye and brain level and has been linked to benefits for vision, cognitive function and other conditions. While marigold flower is positioned as the only crop from which lutein is extracted from and commercialized, microalgae are proposed as an alternative with several advantages over this terrestrial crop. The main barrier to scaling up lutein production from microalgae to the commercial level is the low productivity compared to the high costs. This review explores strategies to enhance lutein production in microalgae by emphasizing the overall productivity over lutein content alone. Evaluation of how culture parameters, such as light quality, nitrogen sufficiency, temperature and even stress factors, affect lutein content and biomass development in batch phototrophic cultures was performed. Overall, the total lutein production remains low under this metabolic regime due to the low biomass productivity of photosynthetic batch cultures. For this reason, we describe findings on microalgal cultures grown under different metabolic regimes and culture protocols (fed-batch, pulse-feed, semi-batch, semi-continuous, continuous). After a careful literature examination, two-step heterotrophic or mixotrophic cultivation strategies are suggested to surpass the lutein productivity achieved in single-step photosynthetic cultures. Furthermore, this review highlights the urgent need to develop technical feasibility studies at a pilot scale for these cultivation strategies, which will strengthen the necessary techno-economic analyses to drive their commercial production.