Chlorella Vulgaris Growth Research Articles

Growth of Chlorella vulgaris during Active Immobilisation in Calcium Alginate

Growth of Chlorella vulgaris during Active Immobilisation in Calcium Alginate

Effects of coexisting nanomaterials on the photodegradation behavior and ecotoxicity of antibiotics in the aqueous

Nanomaterials (NPs) and antibiotics, as two emergent pollutants, forms a complex contamination through their interaction, potentially causing adverse effects on the organism. This study systematically examined the influence of two NPs (CuO NPs and carbon nanotubes, CNTs) on the photodegradation behavior of tetracycline (TC) and their combined toxic effects on Chlorella vulgaris. The results showed that CuO NPs significantly accelerated TC photodegradation compared to CNTs, increasing the TC photodegradation rate constant by187.6%. Electron spin resonance (ESR) indicated that under the coexistence of CuO NPs or CNTs, 1O2、O2•- and •OH were the main active species promoting TC photodegradation. Probe and quenching experiments confirmed the predominant role of O2•- and 1O2 in the presence of CuO NPs and CNTs. Additionally, three possible TC photodegradation pathways were proposed for the coexistence of CuO NPs and CNTs. In the Chlorella vulgaris growth inhibition experiment, the combined toxicity of CuO NPs or CNTs and TC was higher than that of individual substance, indicating significant synergistic effects, especially with the combination of CNTs and TC. This study provides a new perspective on accurately assessing the environmental behaviors and risks when NPs and antibiotics coexist.

Maximizing resource efficiency with dairy and municipal wastewater as nutrient media for Chlorella vulgaris with simultaneous CO2 sequestration

Addressing critical environmental challenges such as greenhouse gas emissions, global warming, and eutrophication demands urgent and innovative solutions. In recent years, microalgae have emerged as a promising avenue for addressing these pressing issues. In this study, the combination of dairy and municipal wastewater is proposed as a culture medium for cultivating microalgae strains capable of sequestering atmospheric CO2. Specifically, the growth of Chlorella vulgaris was investigated using Bold's basal medium, along with varying concentrations of municipal and dairy wastewater, both with and without CO2 supplementation, to assess their CO2 capture potential. Concurrently, the efficiency of chemical oxygen demand (COD), total nitrogen, potassium, and phosphate removal from the wastewater was evaluated. Additionally, the combination of wastewater media with CO2 supplementation yielded the highest CO2 uptake rates, indicating the feasibility of simultaneous CO2 capture during microalgae cultivation. Media composition with 25% municipal wastewater: 75% dairy wastewater supplemented with CO2 demonstrated superior COD elimination with a higher percentage of nutrient removal from wastewater compared to other wastewater proportions. The nutrient removal capacity of aforementioned media also comes in line with high CO2 sequestration rate (13.57 mg L−1 h−1). These findings underscore the potential of utilizing wastewater from diverse sources as a viable culture medium for microalgae cultivation, facilitating concurrent CO2 capture and wastewater treatment.

Computational fluid dynamics (CFD) simulation modeling for the cultivation of microalgal monoculture in axenic enclosed bubble column photobioreactor (BCPBR)

Researchers are more concerned with axenic-enclosed PBRs, where there is less or no chance of contamination during the production of biochemical and highly valuable metabolites, and monocultures of microalgae are being grown more frequently. It is a closed, manufactured vessel that aids in the photosynthesis of microalgal cells using artificial light or sunlight as the energy source. In this study, the bubble column PBR (BCPBR) was selected because it possessed some advantages over other PBRs for the growth of Chlorella vulgaris. The BCPBR system prevents contact between the enclosed microalgal cells and the environment, allowing the culturing of microalgae species that are difficult to grow in open pond systems. To compare BCPBR performance quantitatively, the efficient mixing expected in BCPBR, as discussed in the literature, was applied to the CFD model. The experimental results observed during the cultivation of C. vulgaris with restaurant wastewater (RWW) in BCPBR clearly showed better mixing, high growth, and improved treatment efficiency. CFD analysis was conducted on the evolution of bubbles in the BCPBR. The Pressure-Implicit with Splitting of Operators (PISO) pressure correction method is used for velocity and pressure coupling. A geo-reconstruct approach is used to construct the interface, and a second-order upwind calculation technique is used to determine the flow parameters. Therefore, CFD simulation in this study will contribute to the following aspects: (i) the volume fraction contours and velocity contours are going to validate the experimental study as the homogenous mixing favors the growth and productivity, (ii) To study how the size of the nozzle and inlet velocity affect the turbulence generated by bubbles in a BCPBR to identify the optimal nozzle size and velocity for the required turbulence.

Regulating active oxygen species and acidity of Ca doped LaMnO3 perovskite catalysts toward efficient n-butylamine oxidation

It is of great significance to design catalysts with high activity, N2 selectivity and stability for nitrogen-containing volatile organic compound (NVOC) catalytic purification. Meanwhile, the role of oxygen species in NVOC catalytic oxidation is still ambiguous. Herein, the oxygen species of La-Mn perovskite catalysts were engineered by doping Ca in La site to elucidate their role in catalytic n-butylamine oxidation. Results demonstrate that La0.9Ca0.1MnO3 exhibits the highest activity owing to its abundant adsorbed oxygen species and high surface area, achieving 100 % n-butylamine conversion (500 ppm; GHSV=60,000 mL g−1h−1) at 200 °C with N2 selectivity of ca. 95 %. More adsorbed oxygen species generated due to Ca doping accelerates the oxidation rate and C-C cleavage of alcohol and acetate intermediates, resulting in higher CO2 yield. Meanwhile, Ca-doping also modifies the acidity of catalysts, which affects the adsorption/desorption process of alkaline substances and yield of nitrogen-containing products. In addition, La0.9Ca0.1MnO3 shows superior stability, high water and SO2 resistance, and acceptable activity for other common NVOCs, demonstrating a promising catalyst for NVOC oxidation even under harsh operation conditions. Furthermore, the toxic effect of pollutant or products during n-butylamine catalytic oxidation on the growth of Chlorella Vulgaris was investigated and results suggest that catalytic oxidation is an environmentally friendly method for NVOC purification. This work sheds light on the relationships among oxygen species, catalyst acidity, and n-butylamine catalytic oxidation, which expand the understanding of rational design of efficient catalyst for NVOC oxidation.

From waste to resource: Effects of digested rotten potato supernatant on the growth, total biomass and nutrient composition of Chlorella vulgaris

A study was carried out to evaluate the growth performance of Chlorella vulgaris, a green microalga, in three different concentrations of digested rotten potato supernatant (DRPS) for 16 days. C. vulgaris was grown in 20 % (T1), 40 % (T2), and 60 % (T3) of the DRPS and at the same time in Bold Basal Medium (BBM) as a control (T4). A significantly highest cell density of C. vulgaris was found in T1 (192.83 ± 1.75 × 105 cells mL−1) in comparison to T2 (136.83 ± 5.58 × 105 cells mL−1), and T3 (99.11 ± 5.38 × 105 cells mL−1) (p < 0.001 for all comparisons) while the cell density at T1 (192.83 ± 1.75 × 105 cells mL−1) and T4 (180.907 ± 4.58 × 105 cells mL−1) did not differ significantly (p = 0.227). Moreover, the mean daily division rate of C. vulgaris was significantly higher in T1 (0.340 ± 0.001 divisions day−1) in comparison to other concentrations of DRPS (p < 0.001 for all comparisons). The maximum value of total biomass (1.07 ± 0.10 g L−1) was found in T1 which was statistically similar to those in T4 and T2. In addition, there was no significant difference between the mean maximum values of chlorophyll-a content and optical density of C. vulgaris in T1 and T4. The highest protein content of 42.67 ± 0.57 % was observed in T4 which was significantly higher than T1 (39.43 ± 1.67 %) (p = 0.027). It is also worth mentioning that there was no significant difference in the crude lipid content of the microalgae grown in T1 (10.06 ± 0.17 %) and T4 (9.88 ± 0.14 %) (p = 0.616). Hence, 20 % DRPS can be used as an alternative culture media of BBM for C. vulgaris with a broad aim to accelerate the sustainable advancement of microalgal production.

Towards a circular economy - Repurposing side streams from the potato processing industry by Chlorella vulgaris

Common wastewater treatment strategies in the food industry do not include efficient remediation strategies for nitrogen, phosphorous and organic carbon. Incorporating microalgae in water treatment plants is rising in popularity because of their high nutrient and trace element uptake driven by light. In this study, four different side streams from an Austrian potato processing company have been screened for their applicability of microalgal cultivation. The side streams were assessed for Chlorella vulgaris growth and their requirement of any additional pretreatment or media supplementation. One side stream specifically, called blanching water II, a stream generated by boiling the potatoes for ease of peeling, turned out very useful to cultivate Chlorella vulgaris and concomitantly remedy the wastewater. Compared to a state-of-the-art cultivation in BG11, cultivating Chlorella vulgaris in blanching water II led to a 45 % increase in specific growth rate of 1.29 day−1 and a 48% increase in biomass productivity to 294.6 mg/L/day, while all nitrogen and phosphate present in the side stream were metabolized. Overall, the results demonstrate that the water remediation process for blanching water II shows vast potential in regard to water purification and waste to value approaches.

Using PyCaret to model Chlorella vulgaris's growth response to salinity and oil contamination for crude oil bioremediation

ABSTRACT Crude oil spills significantly impact aquatic ecosystems, necessitating innovative remediation strategies. Microalgae-based bioremediation, particularly with Chlorella vulgaris, offers a promising solution. This study introduces a novel framework that evaluates the combined effects of selected environmental stressors on microalgal adaptability, advancing beyond traditional isolated factor analyses. By integrating a factorial experimental design with a machine learning approach using PyCaret AutoML and SHAP values, we provide a detailed examination of how crude oil concentration, salinity, and exposure duration affect C. vulgaris growth. The Extra Trees Regressor model emerged as highly accurate in predicting biomass concentration, a crucial adaptability indicator, achieving an MAE of 0.0202, RMSE of 0.029, and an R² of 0.8875. SHAP analysis highlighted salinity and crude oil as significant growth influencers, with exposure duration playing a minor role. Notably, C. vulgaris exhibited more sensitivity to salinity than to crude oil, indicating potential high-salinity challenges but also a strong tolerance to oil pollutants. These findings enhance our understanding of microalgal responses in polluted environments and suggest improved bioremediation approaches for saline waters affected by oil spills, leveraging the synergy of environmental factors and machine learning insights.

Bioconversion of Furanic Compounds by Chlorella vulgaris-Unveiling Biotechnological Potentials.

Lignocellulosic biomass is abundant on Earth, and there are multiple acidic pretreatment options to separate the cellulose, hemicellulose, and lignin fraction. By doing so, the fermentation inhibitors 5-Hydroxymethylfurfural (HMF) and furfural (FF) are produced in varying concentrations depending on the hydrolyzed substrate. In this study, the impact of these furanic compounds on Chlorella vulgaris growth and photosynthetic activity was analyzed. Both compounds led to a prolonged lag phase in Chlorella vulgaris growth. While the photosynthetic yield Y(II) was not significantly influenced in cultivations containing HMF, FF significantly reduced Y(II). The conversion of 5-Hydroxymethylfurfural and furfural to 5-Hydroxymethyl-2-Furoic Acid and 2-Furoic Acid was observed. In total, 100% of HMF and FF was converted in photoautotrophic and mixotrophic Chlorella vulgaris cultivations. The results demonstrate that Chlorella vulgaris is, as of now, the first known microalgal species converting furanic compounds.

Enhancing immobilized Chlorella vulgaris growth with novel buoyant barium alginate bubble beads

Microalgae immobilization in alginate beads shows promise for biomass production and water pollution control. However, carrier instability and mass transfer limitations are challenges. This study introduces buoyant barium alginate bubble beads (BABB), which offer exceptional stability and enhance Chlorella vulgaris growth. In just 12 days, compared to traditional calcium alginate beads, BABB achieved a 20 % biomass increase while minimizing cell leakage and simplifying harvesting. BABB optimization involved co-immobilization with BG-11 medium, enrichment of CO2 in internal bubbles, and the integration of Fe nanoparticles (FeNPs). In the open raceway pond reactor, these optimizations resulted in a 39 % increase in biomass over 7 days compared to the unoptimized setup in closed flasks. Furthermore, enhancements in pigment and organic matter production were observed, along with improved removal of ammonia nitrogen and phosphate. These results highlight the overall advantages of BABB for microalgae immobilization, offering a scientific foundation for their effective utilization.

The responses and tolerance of photosynthetic system in Chlorella vulgaris to the pharmaceutical pollutant carbamazepine

Screening for sensitive toxicological indicators and understanding algal tolerance to pharmaceutical contaminants (PhCs) are essential for assessing PhCs risk and their removal by microalgae. Carbamazepine (CBZ) showed adverse effects on microalgae, but the specific toxicity mechanisms on the most sensitive algal photosynthetic system (PS) remain limited. This study delved into the impact of CBZ exposure on the growth, cell viability, pigment content, and PS of Chlorella vulgaris. The findings revealed a notable inhibition of C. vulgaris growth by CBZ, with an IC50 value of 27.2 mg/L at 96 h. CBZ exposure induced algal membrane damage and cell viability. Intriguingly, CBZ drastically diminished intracellular pigment levels, notably showing "low promotion and high inhibition" of chlorophyll b (Chl b) by 72 h. Moreover, the study identified a decreased number of active reaction centers (RCs) within algal PSII alongside inhibited electron transport from QA to QB on the PSII receptor side, leading to PSII disruption. As an adaptive response to CBZ stress, C. vulgaris stimulated its Chl b synthesis, increased non-photochemical quenching (NPQ), and adapted its tolerance to bright light. Additionally, the alga attempted to compensate for the CBZ-induced reduction in electron transfer efficiency at the PSII receptor side and light energy utilization by increasing its electron transfer from downstream. Principal component analysis (PCA) further verified that the parameters on non-photochemical dissipation, electron transport, and integrative performance were the most sensitive algal toxicological indicators for CBZ exposure, and algal PS has energy protection capability through negative feedback regulation. However, prolonged exposure to high doses of CBZ will eventually result in permanent damage to the algal PS. Hence, attention should be paid to the concentration of CBZ in the effluent and the exposure time, while methods to mitigate algal photodamage should be appropriately sought for algal treatment of dense effluents.

Parametric Study of the Effect of Increased Magnetic Field Exposure on Microalgae Chlorella vulgaris Growth and Bioactive Compound Production

Parametric Study of the Effect of Increased Magnetic Field Exposure on Microalgae Chlorella vulgaris Growth and Bioactive Compound Production

Optimal growth conditions to enhance Chlorella vulgaris biomass production in indoor phyto tank and quality assessment of feed and culture stock

Commercial microalgae cultivation is a dynamic field with ongoing efforts to improve efficiency, reduce costs, and explore new applications. We conducted a study to examine how different light exposure periods affect Chlorella vulgaris's growth. We employed a Phyto tank batch system of approximately 3.5 L with LED light control, controlled airflow, and sterilized bags, maintained at 22.0 ± 2.0 °C indoors. Various methods, including spectrophotometry, and cell counter were employed to monitor Chlorella vulgaris growth under different light exposure cycles. Additionally, quality analysis as feed source was employed by proximate, amino acid, beta-glucan, and microbial content analysis. The results revealed significant variations in C. vulgaris biomass production based on light exposure duration. Notably, the 16:8-h light-dark photoperiod exhibited the highest biomass concentration, reaching 6.48 × 107 ± 0.50 cells/mL with an optical density (OD) of 1.165 absorbance at 682 nm. The 12:12-h light-dark photoperiod produced the second-highest biomass concentration, with 2.305 × 106 ± 0.60 cells/mL at an OD of 0.489. Proximate analysis of dry algae powder revealed low lipid content (0.48 %), high protein content (37.61 %), variable ash concentration (average 10.75 %), and a significant carbohydrate fraction (51.16 %) during extended daylight and shorter dark periods. Amino acid analysis identified nine essential amino acids, with glutamic acid being the most abundant (17.7 %) and methionine the least (0.4 %). Furthermore, quality analysis and microbiological assays demonstrated that the C. vulgaris biomass is well-suited for fish and livestock use as a feed source and possibility as human nutraceuticals. These findings can be considered more environmentally friendly and ethically sound due to the absence of genetic modification.

Improving Microalgae Feasibility Cultivation: Preliminary Results on Exhausted Medium Reuse Strategy

Although microalgae exploitation is very promising, process sustainability is undermined by biomass production and harvesting. Among the various bottlenecks of the production process, particular attention should be paid to the water footprint. Indeed, a huge volume of water is required in microalgae production. Water reuse can support both the water footprint and medium cost reduction, saving water and unconverted substrates. The present study reports preliminary results regarding the utilization of a water reuse strategy for two Chlorophyta microalgae under batch conditions. Growth parameters and chlorophyll content are monitored and the optimal amount of reused medium is assessed. The results show that 70% of the medium can be reused with no loss of specific growth rate and chlorophyll fraction for Pseudococcomyxa simplex in three consecutive batch cultivations. By contrast, a significant decline in Chlorella vulgaris growth was observed after the first cultivation in reused medium, across all tested conditions.

A growth phase analysis on the influence of light intensity on microalgal stress and potential biofuel production

It is urgent to find alternatives to fossil fuels to mitigate the effects of climate change. High light intensity during microalgal growth can promote lipid and carbohydrate accumulation (feedstocks to biofuels). However, more research needs to be done concerning the effect of this factor on microalgal productivity and biochemical composition from a growth phase point of view. This study evaluated the impact of high light intensities (291, 527 and 1107 μmol m−2 s−1) on Chlorella vulgaris growth in synthetic wastewater, nutrient assimilation and biochemical composition in late-exponential and stationary growth phases. Under the highest light intensity, a higher growth rate (0.54 ± 0.02 d−1) and maximum productivity (166 ± 13 mg/L d−1) were achieved. Also, nutrient removal efficiencies at the late-exponential phase reached 97.0 ± 0.2 % and 98.2 ± 0.1 % for nitrogen and phosphorus, respectively. Under all light intensities tested at the stationary phase, microalgae also achieved high nutrient removal efficiencies (>95 %). Extreme light intensity (1107 μmol m−2 s−1) boosted the accumulation of carbohydrates (30 ± 3 % w/w) and lipids (13.0 ± 0.4 % w/w). Under lower light intensities, the content of these compounds did not change significantly between growth phases. The chlorophyll content decreased with the increasing light intensity and increasing exposure time. Exposing C. vulgaris to high light intensity seems promising to simultaneously treat wastewater and boost lipid and carbohydrate accumulation, which can be used for biofuel production.

Prediction of acute toxicity for Chlorella vulgaris caused by tire wear particle-derived compounds using quantitative structure-activity relationship models

Tire wear particles (TWPs) enter aquatic ecosystems through various pathways, such as rainwater and urban runoff. Additives in TWPs can harm aquatic organisms in these ecosystems. Therefore, it is essential to investigate their toxicity to aquatic organisms. In our study, we initially recorded the median effective concentrations of 21 TWP-derived compounds on Chlorella vulgaris growth, ranging from 0.04 to 8.60 mg/L. Subsequently, through an extensive review of the literature, we incorporated 112 compounds with specific toxicity endpoints to construct the QSAR model using genetic algorithm and multiple linear regression techniques, followed by the construction of the consensus model and the quantitative read-across structure-activity relationship (q-RASAR) model. Meanwhile, we employed rigorous internal and external validation measures to assess the performance of the model. The results indicated that the developed q-RASAR model exhibited strong adaptation, robustness, and reliable prediction, with q-RASAR indicators of Q2LOO = 0.7673, R2tr = 0.8079, R2test = 0.8610, Q2Fn = 0.8285−0.8614, and CCCtest = 0.9222. Based on an external dataset containing 128 emerging TWP-derived compounds, the model's applicability domain coverage was 90.6 %. The q-RASAR model predicted that the structure of diphenylamine was associated with higher toxicity, possibly liked to the SpMax2_Bhm and LogBCF descriptors. The established model reliably provides prediction and fills a critical data gap. These findings highlight the potential risks posed by emerging TWP-derived compounds to aquatic organisms.

Combined toxic effects of polystyrene nanoplastics and lead on Chlorella vulgaris growth, membrane lipid peroxidation, antioxidant capacity, and morphological alterations.

In recent years, there has been a significant rise in the utilization of amino-functionalized polystyrene nanoplastics (PS-NH2). This surge in usage can be attributed to their exceptional characteristics, including a substantial specific surface area, high energy, and strong reactivity. These properties make them highly suitable for a wide range of industrial and medical applications. Nevertheless, there is a growing apprehension regarding their potential toxicity to aquatic organisms, particularly when considering the potential impact of heavy metals like lead (Pb) on the toxicity of PS-NH2. Herein, we examined the toxic effects of sole PS-NH2 (90nm) at five concentrations (e.g., 0, 0.125, 0.25, 0.5, and 1mg/L), as well as the simultaneous exposure of PS-NH2 and Pb2+ (using two environmental concentrations, e.g., 20μg/L for Pb low (PbL) and 80μg/L for Pb higher (PbH)) to the microalga Chlorella vulgaris. After a 96-h exposure, significant differences in chlorophyll a content and algal growth (biomass) were observed between the control group and other treatments (ANOVA, p < 0.05). The algae exposed to PS-NH2, PS-NH2 + PbL, and PS-NH2 + PbH treatment groups exhibited dose-dependent toxicity responses to chlorophyll a content and biomass. According to the Abbott toxicity model, the combined toxicity of treatment groups of PS-NH2 and PbL,H showed synergistic effects. The largest morphological changes such as C. vulgaris' size reduction and cellular aggregation were evident in the medium treated with elevated concentrations of both PS-NH2 and Pb2+. The toxicity of the treatment groups followed the sequence PS-NH2 < PS-NH2 + PbL < PS-NH2 + PbH. These results contribute novel insights into co-exposure toxicity to PS-NH2 and Pb2+ in algae communities.

Comparison of Different Pretreatment Processes Envisaging the Potential Use of Food Waste as Microalgae Substrate.

A significant fraction of the food produced worldwide is currently lost or wasted throughout the supply chain, squandering natural and economic resources. Food waste valorization will be an important necessity in the coming years. This work investigates the ability of food waste to serve as a viable nutritional substrate for the heterotrophic growth of Chlorella vulgaris. The impact of different pretreatments on the elemental composition and microbial contamination of seven retail food waste mixtures was evaluated. Among the pretreatment methods applied to the food waste formulations, autoclaving was able to eliminate all microbial contamination and increase the availability of reducing sugars by 30%. Ohmic heating was also able to eliminate most of the contaminations in the food wastes in shorter time periods than autoclave. However, it has reduced the availability of reducing sugars, making it less preferable for microalgae heterotrophic cultivation. The direct utilization of food waste containing essential nutrients from fruits, vegetables, dairy and bakery products, and meat on the heterotrophic growth of microalgae allowed a biomass concentration of 2.2 × 108 cells·mL-1, being the culture able to consume more than 42% of the reducing sugars present in the substrate, thus demonstrating the economic and environmental potential of these wastes.

Photoheterotroph improved the growth and nutrient levels of Chlorella vulgaris and the related molecular mechanism

Microalgae are rich in fatty acids, proteins, and other nutrients, which have gained the general attention of researchers all over the world. For the development of Chlorella vulgaris in food and feed industry, this study was conducted to investigate the differences in C. vulgaris’ growth and nutritional components under different culture conditions (autotrophic, heterotrophic, photoheterotrophic) and the internal factors through cell counting in combination with transcriptome and nutrient analyses. The results showed that, under the photoheterotrophic condition, Chlorella’s growth and the contents of lipid and protein were significantly higher than that under the heterotrophic condition, and the moisture content was lower than that under the heterotrophic condition. The saturated fatty acid content under the photoheterotrophic condition was the lowest, while the polyunsaturated fatty acid content was significantly higher than those under the other two conditions. There were 46,583 differentially expressed genes (DEGs), including 33,039 up-regulated DEGs (70.93%) and 13,544 down-regulated DEGs (29.07%), under the photoheterotrophic condition in comparison with the autotrophic condition. The fold change between the two conditions of samples of up-regulated genes was higher than that of the down-regulated genes. The KEGG enrichment showed that the up-regulated DEGs in the photoheterotrophic condition were significantly enriched in 5 pathways, including protein processing in endoplasmic reticulum pathway, photosynthesis pathway, photosynthesis-antenna protein pathway, endocytosis pathway, and phosphonate and phosphinate metabolism pathway. DEGs related to fatty acid metabolic pathways were significantly enriched in the fatty acid biosynthesis pathway and the biosynthesis of unsaturated fatty acid pathway. The qPCR analysis showed that the expression pattern of the selected genes was consistent with that of transcriptome analysis. The results of this study lay a theoretical foundation for the large-scale production of Chlorella and its application in food, feed, and biodiesel.Key points• Nutrient levels under photoheterotrophic condition were higher than other conditions.• Six important pathways were discovered that affect changes in nutritional composition.• Explored genes encode important enzymes in the differential metabolic pathways.

Development of a new sterilization method for microalgae media using calcium hypochlorite as the sterilant.

Calcium hypochlorite (Ca(ClO)2), which can be stably stored in powder form for a long period, is widely used as a disinfectant in various fields. A new sterilization process was developed in the present study, where a microalgal medium was sterilized using 0.02% Ca(ClO)2, followed by complete neutralization of the Ca(ClO)2 within 8h through catalytic reaction of an MnCl2-Na2EDTA complex and a synergistic effect of glucose. When comparing the growth of Chlorella vulgaris in the autoclaved medium, a 2.65 times greater maximum cell growth was observed in cells grown in the medium prepared by treatment of Ca(ClO)2. This result indicates that denaturation of the medium by heat can hinder the growth of some microorganisms. In the case of cultivation of Euglena gracilis, successful culture growth was achieved without growth inhibition or contamination on a medium prepared in the same manner.