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Microalgal-indigenous bacterial wastewater treatment (MBWT) emerges as a promising approach for the concurrent removal of nitrogen (N) and phosphorus (P). Despite its potential, the prevalent use of MBWT in batch systems limits its broader application. Furthermore, the success of MBWT critically depends on the stable self-adaptation and synergistic interactions between microalgae and indigenous bacteria, yet the underlying biological mechanisms are not fully understood. Here we explore the viability and microbial dynamics of a continuous flow microalgae-indigenous bacteria advanced wastewater treatment system (CFMBAWTS) in processing actual secondary effluent, with a focus on varying hydraulic retention times (HRTs). The research highlights a stable, mutually beneficial relationship between indigenous bacteria and microalgae. Microalgae and indigenous bacteria can create an optimal environment for each other by providing essential cofactors (like iron, vitamins, and indole-3-acetic acid), oxygen, dissolved organic matter, and tryptophan. This collaboration leads to effective microbial growth, enhanced N and P removal, and energy generation. The study also uncovers crucial metabolic pathways, functional genes, and patterns of microbial succession. Significantly, the effluent NH4+-N and P levels complied with the Chinese national Class-II, Class-V, Class-IA, and Class-IB wastewater discharge standards when the HRT was reduced from 15 to 6 h. Optimal results, including the highest rates of CO2 fixation (1.23 g L−1), total energy yield (32.35 kJ L−1), and the maximal lipid (33.91%) and carbohydrate (41.91%) content, were observed at an HRT of 15 h. Overall, this study not only confirms the feasibility of CFMBAWTS but also lays a crucial foundation for enhancing our understanding of this technology and propelling its practical application in wastewater treatment plants.

Abstract Background Chlorella sorokiniana FZU60 is a promising lutein producing microalga. A mixotrophy/photoautotrophy two-stage strategy can achieve high biomass concentration at stage 1 and high lutein content at stage 2, leading to excellent lutein production efficiency in C. sorokiniana FZU60. However, the underlying molecular mechanisms are still unclear, restraining the further improvement of lutein production. Results In this study, physiological and biochemical analysis revealed that photochemical parameters (Fv/Fm and NPQ) and photosynthetic pigments contents increased during the shift from mixotrophy to photoautotrophy, indicating that photosynthesis and photoprotection enhanced. Furthermore, transcriptomic analysis revealed that the glyoxylate cycle and TCA cycle were suppressed after the shift to photoautotrophy, leading to a decreased cell growth rate. However, the gene expression levels of photosynthesis, CO2 fixation, autophagy, and lutein biosynthesis were upregulated at the photoautotrophy stage, demonstrating that microalgal cells could obtain more precursor to synthesize lutein for enhancing photosynthesis and reducing reactive oxygen species. Conclusions The findings help to elucidate the molecular mechanisms for high lutein production efficiency of C. sorokiniana FZU60 under the mixotrophy/photoautotrophy strategy, identify key functional genes responsible for lutein biosynthesis, and shed light on further improvement of lutein production by genetic or metabolic engineering in future studies.

Highlights Machine learning, techno-economic analysis, and life cycle analysis are imperative for various conversion approaches of high availability and low utilization biomass (HALUB). The conversion of HALUB to sustainable energy and materials has a positive consequence on mitigating climate change and building a green future. Microfluidic and micro/nanomotors-powered sustainable materials are of high potential for advanced applications.

Conventional biological treatment usually cannot achieve the same high water quality as advanced treatment when conducted under varied temperatures. Here, satisfactory wastewater treatment efficiency was observed in a microalgae-bacteria consortia (MBC) over a wide temperature range because of the predominance of microalgae. Microalgae contributed more toward wastewater treatment at low temperature because of the unsatisfactory performance of the accompanying bacteria, which experienced cold stress (e.g., bacterial abundance below 3000 sequences) and executed defensive strategies (e.g., enrichment of cold-shock proteins). A low abundance of amoA-C and hao indicated that conventional nitrogen removal was replaced through the involvement of microalgae. Diverse heterotrophic bacteria for nitrogen removal were identified at medium and high temperatures, implying this microbial niche treatment contained diverse flexible consortia with temperature variation. Additionally, pathogenic bacteria were eliminated through microalgal photosynthesis. After fitting the neutral community model and calculating the ecological niche, microalgae achieved a maximum niche breadth of 5.21 and the lowest niche overlap of 0.38, while the accompanying bacterial community in the consortia were shaped through deterministic processes. Finally, the maximum energy yield of 87.4 kJ L−1 and lipid production of 1.9 g L−1 were achieved at medium temperature. Altogether, this study demonstrates that advanced treatment and energy reclamation can be achieved through microalgae-bacteria niche strategies.

Efficient wastewater treatment accompanied by sustainable “nutrients/pollutants waste-wastewater-resources/energy nexus” management is acting as a prominent and urgent global issue since severe pollution has occurred increasingly. Diverting wastes from wastewater into the value-added microalgal-biomass stream is a promising goal using biological wastewater treatment technologies. This review proposed an idea of upgrading the current microalgal wastewater treatment by using immobilized microalgal system. Firstly, a systematic analysis of microalgal immobilization technology is displayed through an in-depth discussion on why using immobilized microalgae for wastewater treatment. Subsequently, the main technical approaches employed for microalgal immobilization and pollutant removal mechanisms by immobilized microalgae are summarized. Furthermore, from high-tech technologies to promote large-scale production and application potentials in diverse wastewater and bioreactors to downstream applications lead upgradation closer, the feasibility of upgrading existing microalgal wastewater treatment into immobilized microalgal systems is thoroughly discussed. Eventually, several research directions are proposed toward the future immobilized microalgal system for microalgal wastewater treatment upgrading. Together, it appears that using immobilization for further upgrading the microalgae-based wastewater treatment can be recognized as an achievable alternative to make microalgal wastewater treatment more realistic. The information and perspectives provided in this review also offer a feasible reference for upgrading conventional microalgae-based wastewater treatment.

Solar-driven desalination systems have been recognized as an effective technology to address the water crisis. Recently, evaporators prepared based on advanced manufacturing technologies have emerged as a promising tool in enhancing ocean energy utilization. In this review, we discussed the thermal conversion, energy flow, salt deposition mechanisms, and design strategies for solar-driven desalination systems, and explored how to improve the desalination performance and energy use efficiency of the systems through advanced manufacturing technologies. In future perspectives, we determined the feasibility of coupling solar-driven solar desalination systems with multi-stage energy utilization systems and emerging artificial intelligence technologies, for which conclusions are given and new directions for future desalination system development are envisioned. Finally, exciting opportunities and challenges in the face of basic research and practical implementation are discussed, providing promising solutions and blueprints for green and novel desalination technologies while achieving sustainable development.

The rapid expansion of both the global economy and the human population has led to a shortage of water resources suitable for direct human consumption. As a result, water remediation will inexorably become the primary focus on a global scale. Microalgae can be grown in various types of wastewaters (WW). They have a high potential to remove contaminants from the effluents of industries and urban areas. This review focuses on recent advances on WW remediation through microalgae cultivation. Attention has already been paid to microalgae-based wastewater treatment (WWT) due to its low energy requirements, the strong ability of microalgae to thrive under diverse environmental conditions, and the potential to transform WW nutrients into high-value compounds. It turned out that microalgae-based WWT is an economical and sustainable solution. Moreover, different types of toxins are removed by microalgae through biosorption, bioaccumulation, and biodegradation processes. Examples are toxins from agricultural runoffs and textile and pharmaceutical industrial effluents. Microalgae have the potential to mitigate carbon dioxide and make use of the micronutrients that are present in the effluents. This review paper highlights the application of microalgae in WW remediation and the remediation of diverse types of pollutants commonly present in WW through different mechanisms, simultaneous resource recovery, and efficient microalgae-based co-culturing systems along with bottlenecks and prospects.

Microbial fuel cells (MFCs) have gained remarkable attention as a novel wastewater treatment that simultaneously generates electricity. The low activity of the oxygen reduction reaction (ORR) remains one of the most critical bottlenecks limiting the development of MFCs. To date, although research on biochar as an electrocatalyst in MFCs has made tremendous progress, further improvements are needed to make it economically practical. Recently, biochars have been considered to be ORR electrocatalysts with developmental potential. In this review, the ORR mechanism and the essential requirements of ORR catalysts in MFC applications are introduced. Moreover, the focus is to highlight the material selection, properties, and preparation of biochar electrocatalysts, as well as the evaluation and measurement of biochar electrodes. Additionally, in order to provide comprehensive information on the specific applications of biochars in the field of MFCs, their applications as electrocatalysts, are then discussed in detail, including the uses of nitrogen-doped biochar and other heteroatom-doped biochars as electrocatalysts, poisoning tests for biochar catalysts, and the cost estimation of biochar catalysts. Finally, profound insights into the current challenges and clear directions for future perspectives and research are concluded.

Wastewater treatment methods based on catalytic technology are receiving more and more attention in recent years. In order to achieve the high-value utilization of agricultural residues, the development of greener, more efficient, and economically favorable catalysts is the focus of this review. The main contents include: (i) discussions around the differences in surface morphology, chemical stability, electrical conductivity, porosity, and the specific surface area of the prepared biochar according to the biochemical composition of different agricultural residues; (ii) through the above comparison, the shortlisting and selection of suitable catalysts for the preparation of agricultural residues to improve the adsorption and catalytic performance; (iii) discussions around the interaction mode and catalytic mechanism between the surface properties of biochar and pollutants; (iv) based upon future needs, discussions around making full use of the distribution of the pyrolysis products of agricultural residue, and designing biochar-based catalysts to achieve the efficient degradation of pollutants in wastewater. The review aims to provide a waste disposal alternative for the alleviation of environmental pollution and the promotion of green chemistry as well as sustainable development.

Lutein is a high-value carotenoid with many human health benefits. Lycopene β- and ε-cyclases (LCYB and LCYE, respectively) catalyze the cyclization of lycopene into distinct downstream branches, one of which is the lutein biosynthesis pathway, via α-carotene. Hence, LCYB and LCYE are key enzymes in lutein biosynthesis. In this study, the coding genes of two lycopene cyclases (CsLCYB and CsLCYE) of a lutein-enriched marine green microalga, Chlorella sorokiniana FZU60, were isolated and identified. A sequence analysis and computational modeling of CsLCYB and CsLCYE were performed using bioinformatics to identify the key structural domains. Further, a phylogenetic analysis revealed that CsLCYB and CsLCYE were homogeneous to the proteins of other green microalgae. Subcellular localization tests in Nicotiana benthamiana showed that CsLCYB and CsLCYE localized in chloroplasts. A pigment complementation assay in Escherichia coli revealed that CsLCYB could efficiently β-cyclize both ends of lycopene to produce β-carotene. On the other hand, CsLCYE possessed a strong ε-monocyclase activity for the production of δ-carotene and a weak ε-bicyclic activity for the production of ε-carotene. In addition, CsLCYE was able to catalyze lycopene into β-monocyclic γ-carotene and ultimately produced α-carotene with a β-ring and an ε-ring via γ-carotene or δ-carotene. Moreover, the co-expression of CsLCYB and CsLCYE in E. coli revealed that α-carotene was a major product, which might lead to the production of a high level of lutein in C. sorokiniana FZU60. The findings provide a theoretical foundation for performing metabolic engineering to improve lutein biosynthesis and accumulation in C. sorokiniana FZU60.

Abstract Co-production of multiple compounds is an efficient approach to enhance the economic feasibility of microalgae-based metabolites production. In this study, Chlorella sorokiniana FZU60 was cultivated under different bioprocess strategies to enhance the co-production of lutein and protein. Results showed that both lutein and protein content (7.72 and 538.06 mg/g, respectively) were highest at the onset of nitrogen deficiency under batch cultivation. Semi-batch III strategy, with 75% microalgal culture replacement by fresh medium, obtained similar content, productivity, and yield of lutein and protein as batch cultivation, demonstrating that it can be used for stable and continuous production. Fed-batch II strategy, feeding with 1/3 modified BG11 medium, achieved super-high lutein and protein yield (28.81 and 1592.77 mg/L, respectively), thus can be used for high-output production. Besides, two-stage strategy, combining light intensity shift and semi-batch cultivation, gained extremely high lutein and protein productivity (15.31 and 1080.41 mg/L/day, respectively), thereby is a good option for high-efficiency production. Moreover, the fed-batch II and two-stage strategy achieved high-quality lutein and protein, thus are promising for the co-production of lutein and protein in C. sorokiniana FZU60 for commercial application.

Aquatic pollution caused by antibiotics poses a significant threat to human health and the ecosystem. Inspired from “Emmental Cheese” that owns lots of natural pores, we here fabricated a hierarchical cheese-like porous Spirulina residue biochar (KSBC) activated by KHCO3 for efficiently boosting the removal of sulfathiazole (STZ). Through learning form nature that the CO2 produced by bacteria can serve as the natural pore maker (like cheese-making), KHCO3 was thus selected as the gas generating agent in this study. The effect of adding KHCO3 on the surface properties of KSBC was comprehensively investigated. Benefiting from the activation, the KSBC with the mass ratio of 2:1 (2K-SBC) possessed the largest specific surface areas (1100 m2 g−1), which was approximately 81 times that of the original (not activated) Spirulina residue biochar (SBC) (13.56 m2 g−1). Moreover, 2K-SBC exhibited the maximum adsorption capacity for STZ (218.4 mg g−1), dramatically higher than the SBC (25.78 mg g−1). The adsorption kinetics and adsorption isotherms exhibited that the adsorption behavior of 2K-SBC for STZ was consistent with the pseudo-second-order and Langmuir models. Additionally, the adsorption thermodynamics revealed that the adsorption of STZ on 2K-SBC was spontaneous and exothermic. The pore-filling and electrostatic interaction were considered the main mechanism for the adsorption of STZ on 2K-SBC, whereas the π-π electron donor-acceptor (EDA) interaction and hydrogen bond would also partially contribute to the adsorption process.

Abstract Background Scenedesmus obliquus belongs to green microalgae and is widely used in aquaculture as feed, which is also explored for lipid production and bioremediation. However, genomic studies of this microalga have been very limited. Cell self-flocculation of microalgal cells can be used as a simple and economic method for harvesting biomass, and it is of great importance to perform genome-scale studies for the self-flocculating S. obliquus strains to promote their biotechnological applications. Results We employed the Pacific Biosciences sequencing platform for sequencing the genome of the self-flocculating microalga S. obliquus AS-6-11, and used the MECAT software for de novo genome assembly. The estimated genome size of S. obliquus AS-6-11 is 172.3 Mbp with an N50 of 94,410 bp, and 31,964 protein-coding genes were identified. Gene Ontology (GO) and KEGG pathway analyses revealed 65 GO terms and 428 biosynthetic pathways. Comparing to the genome sequences of the well-studied green microalgae Chlamydomonas reinhardtii, Chlorella variabilis, Volvox carteri and Micractinium conductrix, the genome of S. obliquus AS-6-11 encodes more unique proteins, including one gene that encodes D-mannose binding lectin. Genes encoding the glycosylphosphatidylinositol (GPI)-anchored cell wall proteins, and proteins with fasciclin domains that are commonly found in cell wall proteins might be responsible for the self-flocculating phenotype, and were analyzed in detail. Four genes encoding both GPI-anchored cell wall proteins and fasciclin domain proteins are the most interesting targets for further studies. Conclusions The genome sequence of the self-flocculating microalgal S. obliquus AS-6-11 was annotated and analyzed. To our best knowledge, this is the first report on the in-depth annotation of the S. obliquus genome, and the results will facilitate functional genomic studies and metabolic engineering of this important microalga. The comparative genomic analysis here also provides new insights into the evolution of green microalgae. Furthermore, identification of the potential genes encoding self-flocculating proteins will benefit studies on the molecular mechanism underlying this phenotype for its better control and biotechnological applications as well.

The existence of continually increasing concentrations of antibiotics in the environment is a serious potential hazard due to their toxicity and persistence. Unfortunately, conventional treatment techniques, such as those utilized in wastewater treatment plants, are not efficient for the treatment of wastewater containing antibiotic. Recently, algae-based technologies have been found to be a sustainable and promising technique for antibiotic removal. Therefore, this review aims to provide a critical summary of algae-based technologies and their important role in antibiotic wastewater treatment. Algal removal mechanisms including bioadsorption, bioaccumulation, and biodegradation are discussed in detail, with using algae-bacteria consortia for antibiotic treatment, integration of algae with other microorganisms (fungi and multiple algal species), hybrid algae-based treatment and constructed wetlands, and the factors affecting algal antibiotic degradation comprehensively described and assessed. In addition, the use of algae as a precursor for the production of biochar is highlighted, along with the modification of biochar with other materials to improve its antibiotic removal capacity and hybrid algae-based treatment with advanced oxidation processes. Furthermore, recent novel approaches for enhancing antibiotic removal, such as the use of genetic engineering to enhance the antibiotic degradation capacity of algae and the integration of algal antibiotic removal with bioelectrochemical systems are discussed. Finally, some based on the critical review, key future research perspectives are proposed. Overall, this review systematically presents the current progress in algae-mediated antibiotic removal technologies, providing some novel insights for improved alleviation of antibiotic pollution in aquatic environments.

Regulated disposal or re-utilization of dewatered sludge is of economic benefits and can avoid secondary contamination to the environment; however, feasible and effective management strategies are still lacking. In this study, a peroxydisulfate/zero-valent iron (PDS-ZVI) system is proposed to destroy proteins in soluble extracellular polymeric substances (S-EPS) and loosely bound EPS (LB-EPS) in anaerobic digested sludge (ADS) to improve the dewaterability. Moreover, ADS derived biochars supported via iron oxides (Fe-ADSBC) were generated by dewatering and thermal annealing. Intriguingly, the iron species was discovered to gradually transform from Fe3O4 to FeO with increased pyrolysis temperatures from 600 to 1000 °C. The manipulated iron species on the biochar can remarkably impact the catalytic activity in PDS activation and degradation of sulfamethazine (SMT). The in situ radical scavenging and capturing tests revealed that the principal reactive oxygen species (ROS) in Fe-ADSBC/PDS system experienced a variation from OH into SO4− at higher annealing temperature (1000 °C). In addition, the carbonaceous ADSBC can promote the catalytic activity of iron oxides by synergistically facilitating the adsorption of reactants and charge transfer through COFe bonds at the interfaces. This study enables the first insights into the properties and catalytic performance of Fe-ADSBC, meanwhile unveils the mechanism, reaction pathways, and environmental impacts of the ultimate transformation products (TPs) from SMT degradation in the Fe-ADSBC/PDS system. The study also contributes to developing value-added green biochar catalysts from bio-wastes towards environmental purification. Keywords: Anaerobic digested sludge, Biochar, Peroxydisulfate, Sulfamethazine

Summary: Biomass-derived carbon quantum dots (CQDs) are attractive to serve as fluorescent nanosensors owing to their superior environmental compatibility and biocompatibility. However, the detection range has been limited, only in partial agreement with the experimental data. Thus, an advanced kinetic model for quantifying the fluorescence quenching over a wide range is on demand. Here, we describe a nanosensor for Fe(Ⅲ) detection in real waters, which is developed via microalgal residue-derived CQDs with an advanced kinetic model. The multiple-order kinetic model is established to resolve the incoherence of previous models and unveil the entire quenching kinetics. The results show that the detection range of Fe(Ⅲ) can reach up to 10 mM in the high detection end. The newly obtained kinetic model exhibits satisfactory fittings, clearly elucidating a dynamic quenching mechanism. This work provides a new insight into CQDs-based detection of heavy metals in real water samples by establishing an innovative multiple-order kinetic model.

Formation of acid mine drainage (AMD) is a widespread environmental issue that has not subsided throughout decades of continuing research. Highly acidic and highly concentrated metallic streams are characteristics of such streams. Humans, plants and surrounding ecosystems that are in proximity to AMD producing sites face immediate threats. Remediation options include active and passive biological treatments which are markedly different in many aspects. Sulfate reducing bacteria (SRB) remove sulfate and heavy metals to generate non-toxic streams. Passive systems are inexpensive to operate but entail fundamental drawbacks such as large land requirements and prolonged treatment period. Active bioreactors offer greater operational predictability and quicker treatment time but require higher investment costs and wide scale usage is limited by lack of expertise. Recent advancements include the use of renewable raw materials for AMD clean up purposes, which will likely achieve much greener mitigation solutions.

Abstract Background Organic carbon sources have been reported to simultaneously increase the growth and lipid accumulation in microalgae. However, there have been no studies of the mixotrophic growth of Porphyridium purpureum in organic carbon media. In this study, three organic carbon sources, glucose, sodium acetate, and glycerol were used as substrates for the mixotrophic growth of P. purpureum. Moreover, a novel trait-based approach combined with Generalized Additive Modeling was conducted to determine the dosage of each organic carbon source that optimized the concentration of cell biomass or fatty acid. Results A 0.50% (w/v) dosage of glucose was optimum for the enhancement of the cell growth of P. purpureum, whereas sodium acetate performed well in enhancing cell growth, arachidonic acid (ARA) and eicosapentaenoic acid (EPA) content, and glycerol was characterized by its best performance in promoting both cell growth and ARA/EPA ratio. The optimum dosages of sodium acetate and glycerol for the ARA concentration were 0.25% (w/v) and 0.38% (v/v), respectively. An ARA concentration of 211.47 mg L−1 was obtained at the optimum dosage of glycerol, which is the highest ever reported. Conclusions The results suggested that a comprehensive consider of several traits offers an effective strategy to select an optimum dosage for economic and safe microalgae cultivation. This study represents the first attempt of mixotrophic growth of P. purpureum and proved that both biomass and ARA accumulation could be enhanced under supplements of organic carbon sources, which brightens the commercial cultivation of microalgae for ARA production.

Abstract Background Oil leakages frequently occur during oil product development and oil transportation. These incidents are a vital factor in water contamination, thus leading to serious environmental destruction. Therefore, superhydrophobic/superoleophilic material is one of the solutions to treat oily wastewater. Results This study aimed to develop a simple, fast and low-cost method to treat oily wastewater by synthesizing a new superhydrophobic/superoleophilic corn straw fiber via conventional impregnation. The corresponding results illustrate that abundant homogeneous silica (SiO2) granules evenly accreted on the surface of the prepared fiber were conducive to high surface roughness. Meanwhile, (Heptadecafluoro-1,1,2,2-tetradecyl) trimethoxysilane, a sort of silane coupling agent, could greatly reduce surface free energy by grafting with SiO2 particles on the corn straw fiber surface. The obtained superhydrophobic/superoleophilic corn straw fiber exhibited a water contact angle of 152° and an oil contact angle of 0° for various oils, strongly demonstrating its considerable application as an oil absorbent that can be applied for oil cleanup. In addition, the prepared fiber displayed a great chemical stability and environmental durability. Conclusions Due to its high absorption capacity and absorption efficiency, the prepared fiber has great potential as a new oil absorbent for treatment of oily water.

Abstract Background With the current rapid economic growth, demands for energy are progressively increasing. Energy shortages have attracted significant attention due to the shrinking availability of non-renewable resources. Therefore, thermal energy storage is one of the solutions that lead to saving of fossil fuels and make systems more cost-effective by the storage of wasted thermal energy. In particular, the application of phase change materials (PCMs) is considered as an effective and efficient approach to thermal energy storage because of the high latent heat storage capacity at small temperature intervals. Nevertheless, leakage problems and low thermal conductivity limit the practical applications of PCMs. Therefore, form-stable phase change materials with high thermal conductivity are urgently needed. Results A novel form-stable composite phase change material was prepared by incorporating PEG into waste sawdust with 5% EG. In the composites, PEG served as a phase change material, while waste sawdust acted as a carrier matrix. EG was added to help increase the thermal conductivity of the composites. The melting temperature of CPCMs-4 with 5% EG was found to be 58.6 °C with a phase change enthalpy of 145.3 kJ/kg, while the solidifying temperature was 48.5 °C with a phase change enthalpy of 131.4 kJ/kg. The thermal conductivity of CPCMs-4 with 5% EG increased by 23.8% compared with that of CPCMs-4. Moreover, no obvious changes in melting, solidifying temperature, or latent heat after 200 heating–cooling cycles were detected. The supercooling extent of CPCMs-4 with 5% EG decreased by 19.2% compared with PEG. The volume change properties and wettability properties of CPCMs-4 with 5% EG are suitable for thermal energy in terms of practical application. Conclusions The prepared composites have excellent thermal and form-stable properties and they can be recognized as potential candidates for thermal energy storage as form-stable composite phase change materials. Using simple impregnation techniques with waste sawdust as a supporting material, this study demonstrates an innovative technology for practically and markedly enhancing the adsorption capacity of phase change materials.

Our environment is facing several serious challenges from energy utilization, such as fossil fuel exhaustion, air pollution, deteriorated atmospheric greenhouse effect, global warming, climate change, etc [...]

Abstract Background The light/dark cycle is one of the most important factors affecting the microalgal growth and lipid accumulation. Biomass concentration and lipid productivity could be enhanced by optimization of light/dark cycles, and this is considered an effective control strategy for microalgal cultivation. Currently, most research on effects of light/dark cycles on algae is carried out under autotrophic conditions and little information is about the effects under mixotrophic cultivation. At the same time, many studies related to mixotrophic cultivation of microalgal strains, even at large scale, have been performed to obtain satisfactory biomass and lipid production. Therefore, it is necessary to investigate cellular metabolism under autotrophic and mixotrophic conditions at different light/dark cycles. Even though microalgal lipid production under optimal environmental factors has been reported by some researchers, the light/dark cycle and temperature are regarded as separate parameters in their studies. In practical cases, light/dark cycling and temperature variation during the day occur simultaneously. Therefore, studies about the combined effects of light/dark cycles and temperature variation on microalgal lipid production are of practical value, potentially providing significant guidelines for large-scale microalgal cultivation under natural conditions. Results In this work, cell growth and lipid accumulation of an oleaginous microalgal mutant, Scenedesmus sp. Z-4, were investigated at five light/dark cycles (0 h/24 h, 8 h/16 h, 12 h/12 h, 16 h/8 h, and 24 h/0 h) in batch culture. The results showed that the optimal light/dark cycle was 12 h/12 h, when maximum lipid productivity rates of 56.8 and 182.6 mg L−1 day−1 were obtained under autotrophic and mixotrophic cultivation, respectively. Poor microalgal growth and lipid accumulation appeared in the light/dark cycles of 0 h/24 h and 24 h/0 h under autotrophic condition. Prolonging the light duration was unfavorable to the production of chlorophyll a and b, which was mainly due to photooxidation effect. Polysaccharide was converted into lipid and protein when the light irradiation time increased from 0 to 12 h; however, further increasing irradiation time had a negative effect on lipid accumulation. Due to the dependence of autotrophically cultured cells on light energy, the light/dark cycle has a more remarkable influence on cellular metabolism under autotrophic conditions. Furthermore, the combined effects of temperature variation and light/dark cycle of 12 h/12 h on cell growth and lipid accumulation of microalgal mutant Z-4 were investigated under mixotrophic cultivation, and the results showed that biomass was mainly produced at higher temperatures during the day, and a portion of biomass was converted into lipid under dark condition. Conclusions The extension of irradiation time was beneficial to biomass accumulation, but not in favor of lipid production. Even though effects of light/dark cycles on autotrophic and mixotrophic cells were not exactly the same, the optimal lipid productivities of Scenedesmus sp. Z-4 under both cultivation conditions were achieved at the light/dark of 12 h/12 h. This may be attributed to its long-term acclimation in natural environment. By combining temperature variation with optimal light/dark cycle of 12 h/12 h, this study will be of great significance for practical microalgae-biodiesel production in the outdoor conditions.

Marine microalgae are regarded as potential feedstock because of their multiple valuable compounds, including lipids, pigments, carbohydrates, and proteins. Some of these compounds exhibit attractive bioactivities, such as carotenoids, ω-3 polyunsaturated fatty acids, polysaccharides, and peptides. However, the production cost of bioactive compounds is quite high, due to the low contents in marine microalgae. Comprehensive utilization of marine microalgae for multiple compounds production instead of the sole product can be an efficient way to increase the economic feasibility of bioactive compounds production and improve the production efficiency. This paper discusses the metabolic network of marine microalgal compounds, and indicates their interaction in biosynthesis pathways. Furthermore, potential applications of co-production of multiple compounds under various cultivation conditions by shifting metabolic flux are discussed, and cultivation strategies based on environmental and/or nutrient conditions are proposed to improve the co-production. Moreover, biorefinery techniques for the integral use of microalgal biomass are summarized. These techniques include the co-extraction of multiple bioactive compounds from marine microalgae by conventional methods, super/subcritical fluids, and ionic liquids, as well as direct utilization and biochemical or thermochemical conversion of microalgal residues. Overall, this review sheds light on the potential of the comprehensive utilization of marine microalgae for improving bioeconomy in practical industrial application.

Microalgae are considered as excellent candidates for bioactive compounds, yet microalgal residues remaining after the extraction of one or two compounds are usually discarded, which is not economical. This study demonstrates the alkaline extraction of proteins from Chlorella pyrenoidosa residue after lipid and pigment extractions, and their functional properties. Single-factor experiments and response surface methodology were used to obtain the optimal conditions for protein extraction. Based on our results, a maximum protein yield of 722.70 mg/g, was obtained under the following extraction conditions: sodium hydroxide concentration 7.90%, extraction temperature 70.00 °C, extraction time 34.80 min, and microalgal residue concentration 8.20 mg/mL. The molecular weight of microalgal residue protein isolate (MRPI) was mainly distributed at the regions of 0.18−0.50 kDa, 0.50−1.50 kDa, and 1.50−5.00 kDa. The essential amino acid content was greater than the values recommended by FAO/WHO standards; a high essential amino acid index value (1.49) was another good indication that MRPI is suitable for human consumption. Moreover, MRPI exhibited excellent emulsifying properties and antioxidant activity, which suggests it may be useful as an emulsifying agent and antioxidant. These findings could improve the extraction methods of functional protein from microalgal residue and add value to microalgae-based bioactive compound production processes.