The growth of microalgae under alkaline conditions ensures an ample supply of CO2 from the atmosphere, with a low risk of crashing due to contamination and predators. The present study investigated the mixotrophic cultivation of two alkaliphilic microalgae (Tetradesmus obliquus and Cyanothece sp.) using cheese whey as an organic carbon source. The variation in cheese whey concentration (0.5-4.5% (v/v)), culture pH (7-11), and NaNO3 concentrations (0-2 gL-1) was evaluated using central composite design in response to biomass productivity and the contents of lipids, total proteins, and soluble carbohydrates. Both investigated microalgae effectively utilized cheese whey as an organic carbon source. The optimum conditions for simultaneously maximizing biomass and lipid productivity in T. obliquus were 3.5% (v/v) whey, pH 10.0, and 0.5 g L-1 NaNO3. Under these conditions, the biomass, lipid, soluble carbohydrate, and protein productivities were 48.69, 20.64, 7.02, and 10.97 mg L-1 day-1, respectively. Meanwhile, Cyanothece produced 52.78, 11.42, 4.31, and 7.89 mg L-1 day-1 of biomass, lipid, carbohydrate, and protein, respectively, at 4.5% (v/v) whey, pH 9.0, and 1.0 g L-1 NaNO3. The lipids produced under these conditions were rich in saturated fatty acids (FAs) and monounsaturated FAs, with no polyunsaturated FAs in both microalgae. Moreover, several biodiesel characteristics were estimated, and results fell within the ranges specified by international standards. These findings indicate that the mixotrophic cultivation of alkaliphilic microalgae could open new avenues for promoting microalgae productivity through low-cost biofuel production.

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.

Heterotrophic-sulfur autotrophic denitrification (HAD) has been proposed to be a prospective nitrogen removal process. In this work, the potential of fermentation liquid (FL) from waste-activated sludge (WAS) as the electron donor for denitrification in the HAD system was explored and compared with other conventional carbon sources. Results showed that when FL was used as a carbon source, over 99% of NO3--N was removed and its removal rate exceeded 14.00 mg N/g MLSS/h, which was significantly higher than that of methanol and propionic acid. The produced sulfate was below the limit value and the emission of N2O was low (1.38% of the NO3--N). Microbial community analysis showed that autotrophic denitrifiers were predominated in the HAD system, in which Thiobacillus (16.4%) was the dominant genus. The economic analysis showed the cost of the FL was 0.062 €/m3, which was 30% lower than that in the group dosed with methanol. Our results demonstrated the FL was a promising carbon source for the HAD system, which could reduce carbon emission and cost, and offer a creative approach for waste-activated sludge resource reuse.

Fucoxanthin is a versatile substance in the food and pharmaceutical industries owing to its excellent antioxidant and anti-obesity properties. Several microalgae, including the haptophyte Pavlova spp., can produce fucoxanthin and are potential industrial fucoxanthin producers, as they lack rigid cell walls, which facilitates fucoxanthin extraction. However, the commercial application of Pavlova spp. is limited owing to insufficient biomass production. In this study, we aimed to develop a mixotrophic cultivation method to increase biomass and fucoxanthin production in Pavlova gyrans OPMS 30543X. The effects of culturing OPMS 30543X with different organic carbon sources, glycerol concentrations, mixed-nutrient conditions, and light intensities on the consumption of organic carbon sources, biomass production, and fucoxanthin accumulation were analyzed. Several organic carbon sources, such as glycerol, glucose, sucrose, and acetate, were examined, revealing that glycerol was well-consumed by the microalgae. Biomass and fucoxanthin production by OPMS 30543X increased in the presence of 10 mM glycerol compared to that observed without glycerol. Metabolomic analysis revealed higher levels of the metabolites related to the glycolytic, Calvin-Benson-Bassham, and tricarboxylic acid cycles under mixotrophic conditions than under autotrophic conditions. Cultures grown under mixotrophic conditions with a light intensity of 100 µmol photons m-2 s-1 produced more fucoxanthin than autotrophic cultures. Notably, the amount of fucoxanthin produced (18.9 mg/L) was the highest reported thus far for Pavlova species. In conclusion, the use of mixotrophic culture is a promising strategy for increasing fucoxanthin production in Pavlova species. KEY POINTS: • Glycerol enhances biomass and fucoxanthin production in Pavlova gyrans • Metabolite levels increase under mixotrophic conditions • Mixotrophic conditions and medium-light intensity are appropriate for P. gyrans.

CO2 absorption-microalgae conversion (CAMC) system is a promising carbon capture and utilization technology. However, the use of HCO3- as a carbon source often led to a slower growth rate of microalgae, which also limited the application of CAMC system. In this study, the assimilation efficiency of HCO3- in CAMC system was improved through mixotrophic, and the potential mechanism was investigated. The HCO3- assimilation efficiency and biomass under mixotrophic were 34.79% and 31.76% higher than that of control. Mixotrophic increased chlorophyll and phycocyanin content, which were beneficial to capture more light energy. The content of ATP and NADPH reached 566.86 μmol/gprot and 672.86 nmol/mgprot, which increased by 31.83% and 27.67% compared to autotrophic. The activity of carbonic anhydrase and Rubisco increased by 18.52% and 22.08%, respectively. Transcriptome showed that genes related to photosynthetic and respiratory electron transport were up-regulated. The synergy of photophosphorylation and oxidative phosphorylation greatly improved energy metabolism efficiency, thus accelerating the assimilation of HCO3-. These results revealed a potential mechanism of promoting the HCO3- assimilation under mixotrophic, it also provided a guidance for using CAMC system to serve carbon neutrality.

Microbially mediated NO3--N and Cr(VI) reduction is being recognized as an eco-friendly and cost-effective remediation strategy. Iron sulfide mineral, as a natural inorganic electron donor, has a strong influence on NO3--N and Cr(VI) transformation, respectively. However, little is known about the simultaneous nitrate and chromium removal performance and underlying mechanism in an iron sulfide mineral-involved mixotrophic biofilter. This study demonstrated that the NO3--N and Cr(VI) removal efficiencies were stable at 62 ± 8% and 56 ± 10%, and most of them were eliminated in the 0-100-mm region of the biofilter. Cr(VI) was reduced to insoluble Cr(III) via microbial and chemical pathways, which was confirmed by the SEM-EDS morphology and the XPS spectra of biofilm and pyrite particles. SO42- was as a main byproduct of pyrite oxidation; however, the bacterial SO42- reduction synchronously occurred, evidenced by the variations of TOC and SO42- concentrations. These results suggested that there were complicated and intertwined biochemical relations between NO3--N/Cr(VI)/SO42-/DO (electron acceptors) and pyrite/organics (electron donors). Further investigation indicated that both the maximal biomass and greatest denitrifiers\' relative abundances in microbial sample S1 well explained why the pollutants were removed in the 0-100-mm region. A variety of denitrifiers such as Pseudoxanthomona, Acidovorax, and Simplicispira were enriched, which probably were responsible for both NO3--N and Cr(VI) removal. Our findings advance the understanding of simultaneous nitrate and chromium removal in pyrite-involved mixotrophic systems and facilitate the new strategy development for nitrate and chromium remediation.

The green microalga Chlamydomonas reinhardtii is a model microorganism for several areas of study. Among the different microalgae species, it presents advantageous characteristics, such as genomes completely sequenced and well-established techniques for genetic transformation. Despite that, C. reinhardtii production is still not easily commercially viable, especially due to the low biomass yield. So far there are no reports of scientometric study focusing only on C. reinhardtii biomass production process. Considering the need for culture optimization, a scientometric research was conducted to analyze the papers that investigated the growth regimes effects in C. reinhardtii cultivation. The search resulted in 130 papers indexed on Web of Science and Scopus platforms from 1969 to December 2022. The quantitative analysis indicated that the photoautotrophic regime was the most employed in the papers. However, when comparing the three growth regimes, the mixotrophic one led to the highest production of biomass, lipids, and heterologous protein. The production of bioproducts was considered the main objective of most of the papers and, among them, biomass was the most frequently investigated. The highest biomass production reported among the papers was 40 g L-1 in the heterotrophic growth of a transgenic strain. Other culture conditions were also crucial for C. reinhardtii growth, for instance, temperature and cultivation process.

There are two major problems in the world, fuel deficiency and environmental pollution by fossil fuels. Microalgae are regarded as one of the most feasible feedstocks for the manufacturing of biofuels and are used in the degradation of fossil fuel spills. The present study was possessed to investigate the ability of green alga Chlorella vulgaris, blue-green alga Synechococcus sp, and its consortium to grow and degrade hydrocarbon such as kerosene (k) with different concentrations (0, 0.5, 1, and 1,5%), and also using algal biomasses to produce biofuel. The algal growth was estimated by optical density (O.D) at 600 nm, pigment contents such as Chlorophyll a,b carotenoid, and dry weight. The kerosene degradation was estimated by FT-IR analysis after and before the cultivation of algae and its consortium. The components of the methanol extract were determined by GC-MS spectroscopy. The results denote the best growth was determined by O.D, algae consortium with 1.5% Kerosene after ten days, meanwhile, the highest dry weight was with C. vulgaris after ten days of cultivation. The FT-IR demonstrated the algae and consortium possessed high efficacy to degrade kerosene. After 15 days of algae cultivation with 1% K, C.vulgaris produced the maximum amount of lipids (32%). The GC-MS profile of methanol extract of two algae and consortium demonstrated that Undecane was presented in high amounts, C.vulgaris (19.9%), Synechococcussp (82.16%), algae consortium (79.51%), and also were presented moderate amounts of fatty acid methyl ester in Synechococcus sp. Overall, our results indicate that a consortium of algae can absorb and remove kerosene from water, and at the same time produce biofuels like biodiesel and petroleum-based fuels.

The present study investigated the effects of different phenol concentrations (200 - 1000 mg L-1) towards Chlorella sp. under different culture conditions (light vs. dark) and NaNO3 concentrations (0 - 0.1 g L-1) using central composite design. Phenol induced hormesis effects on the algal growth and cellular metabolites. Nitrate was identified as a crucial factor for promoting the uptake of phenol by Chlorella cells, while light was a limiting factor for growth, but the phyco-toxicity of phenol was decreased in the dark. The pigment contents were generally increased in the treated cells to protect against the oxidative phenol stress. The incorporation of 200 mg L-1 phenol and 0.05 g L-1 NaNO3 to the illuminated cells markedly promoted biomass and lipid contents to 0.22 g L-1 and 26.26% w/w, which was 44 and 112% higher than the phenol-less control, respectively. Under the same conditions, the increase of phenol concentration to 600 mg L-1, the protein contents were increased to 18.59% w/w. Conversely, the algal cells were able to accumulate more than 60% w/w of soluble carbohydrates under dark conditions at 600 mg L-1 of phenol. Nitrate replete conditions stimulated lipid accumulation at the expense of protein biosynthesis. Furthermore, most of the treatments showed an increase of H2O2 and malonaldehyde contents, especially for the illuminated cells. However, catalase activity tended to increase under dark conditions, especially at low phenol and nitrate concentrations. This study is valuable in indicating the effects of phenol on microalgae by exploiting response surface methodology, which can be applied as a powerful tool in growth monitoring and toxicity assessment.

At deep-sea hydrothermal vents, sulfur oxidation and iron oxidation are of the highest importance to microbial metabolisms, which are thought to contribute mainly in chemolithoautotrophic groups. In this study, 17 mixotrophic neutrophilic thiosulfate- and iron-oxidizing bacteria were isolated from hydrothermal fields on the Carlsberg Ridge in the Indian Ocean, nine to the γ-proteobacteria (Halomonas (4), Pseudomonas (2), Marinobacter (2), and Rheinheimera (1)), seven to the α-proteobacteria (Thalassospira, Qipengyuania, Salipiger, Seohaeicola, Martelella, Citromicrobium, and Aurantimonas), and one to the Actinobacteria (Agromyces), as determined by their 16S rRNA and genome sequences. The physiological characterization of these isolates revealed wide versatility in electron donors (Fe(II) and Mn(II), or thiosulfate) and a variety of lifestyles as lithotrophic or heterotrophic, microaerobic, or anaerobic. As a representative strain, Pseudomonas sp. IOP_13 showed its autotrophic gowth from 105 cells/ml to 107 cells/ml;carbon dioxide fixation capacity with the δ13CVPDB in the biomass increased from -27.42‱ to 3460.06‱; the thiosulfate-oxidizing ability with produced SO42- increased from 60 mg/L to 287 mg/L; and the iron-oxidizing ability with Fe(II) decreased from 10 mM to 5.2 mM. In addition, iron-oxide crust formed outside the cells. Gene coding for energy metabolism involved in possible iron, manganese, and sulfur oxidation, and denitrification was identified by their genome analysis. This study sheds light on the function of the mixotrophic microbial community in the iron/manganese/sulfur cycles and the carbon fixation of the hydrothermal fields.