Chlorella Products Research Articles

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.

Method Improvement for Quantitative Analysis of Chlorophyll Degradation Compounds, Including Pheophorbide, in Chlorella Products

An official analytical method for chlorophyll degradation compounds, including pheophorbide, in chlorella products, is described in notification Kanshoku No. 99 (May 8, 1981). However, this method has several operational issues, such as the formation of emulsion during liquid-liquid partitioning. Additionally, impurities present in the reagents (sodium sulfate decahydrate or anhydrous sodium sulfate) used to prepare saturated sodium sulfate solution can degrade pheophorbide and other related compounds, resulting in a significant decrease in analytical values. In this study, we thoroughly examined each step of the official method to enhance the operability and develop an alternative method that eliminates the need for saturated sodium sulfate solution. The developed method was evaluated for pheophorbide a and pyropheophorbide a at 100 mg%. Satisfactory analytical performance was achieved with trueness of 100% for pheophorbide a and 90% for pyropheophorbide a, and relative standard deviations of intra- and inter-day precision below 5% for both compounds. The proposed method is considered suitable for regulatory analysis of chlorophyll degradation compounds and would be useful for quality control of chlorella products.

Evaluation of Technical and Economic Indicators for the Production Process of Microalgae Lipids Considering CO2 Capture of a Thermoelectric Plant and Use of Piggery Wastewater

Microalgae are highly studied microorganisms for the production of high-value products due to their high content of proteins, lipids, carbohydrates, and chlorophyll. These compounds are refined to obtain profitable industrial products. This article analyzes the lipid production of Chlorella sp. biomass, considering 18 scenarios for its production, with 9 of these being partially supplemented with swine wastewater. A 1 ha area was considered for biomass cultivation, primary and secondary biomass harvesting, and lipid extraction. Using simulation in the software SuperPro Designer v10, parameters such as CO2 capture (from a thermoelectric power plant), freshwater consumption, wastewater consumption, energy consumption, and unit production cost were evaluated. The results show that the production cost is high, ranging from 836.9 US $/kg to 1131.5 US $/kg of produced lipids, with a maximum CO2 capture of 454 kg of CO2/kg of lipids. The use of wastewater reduces the production cost by approximately 10%. The evaluation of technical and economic parameters allows us to identify bottlenecks and implement strategies to reduce production costs.

Valorization of nutrients from fruit residues for the growth and lipid production of Chlorella sp.: A vision of the circular economy in Peru

Valorization of nutrients from fruit residues for the growth and lipid production of Chlorella sp.: A vision of the circular economy in Peru

Responses of Chlorella vulgaris to the native bacteria in real wastewater: Improvement in wastewater treatment and lipid production

Alga–bacterium interaction can improve wastewater treatment efficiency. To unravel the mystery of the interaction between microalgae and bacteria in wastewater, mono-cultures and co-cultures of Chlorella vulgaris and native bacteria in pretreated biochemical wastewater from landfill leachate were investigated. The results showed that the microalgae selected dominant commensal bacteria, creating a further reduction in species richness for the co-culture, which in turn aids in the dominant commensal bacteria's survival, thereby enhancing algal and bacterial metabolic activity. Strikingly, the lipid productivity of Chlorella in co-culture — namely 41.5 mg/L·d — was 1.4 times higher than in algal monoculture. Additionally, pollutant removal was enhanced in co-cultures, attributed to the bacterial community associated with pollutants' degradation. Furthermore, this study provides an important advance towards observations on the migration and transformation pathways of nutrients and metals, and bridges the gap in algal–bacterial synergistic mechanisms in real wastewater, laying the theoretical foundation for improving wastewater treatment.

Enhancing Lipid Production of Chlorella sp. by Mixotrophic Cultivation Optimization

Mixotrophic microalgal cultivation can utilize CO2 and organic carbon sources. This study optimized the cultivation nitrogen source (peptone, urea, yeast extract, NH4Cl, (NH4)2SO4, NH4NO3, NaNO3 and KNO3), carbon combination (glucose, glycerol, sucrose), (NH4)2SO4 nitrogen source and pH (6–11) in a local microalgae species with three characteristics (high pH-resistant, high growth rate and high lipid content). Chlorella sp. G3H3-1-2 biomass production and lipid accumulation were estimated using the fatty acid methyl esters (FAMEs) concentration. The Chlorella sp. G3H3-1-2 FAME content was strongly influenced by the carbon, nitrogen sources and pH variations. The pH ranged from 6.0 to 8.0, which produced the highest specific growth rate of 1.22 day−1 for Chlorella sp. G3H3-1-2 while using glucose as the single carbon source. However, the highest total FAMEs content of 59% in the Chlorella sp. G3H3-1-2 biomass of 1.75 g/L was obtained while using the combination of 1 g-glucose/L as the carbon source and 0.2 g-(NH4)2SO4/L as the nitrogen source at the high pH value of 10.

A Low-Cost Fertilizer Medium Supplemented with Urea for the Lutein Production of Chlorella sp. and the Ability of the Lutein to Protect Cells against Blue Light Irradiation.

This study aimed to investigate the use of organic fertilizers instead of modified f/2 medium for Chlorella sp. cultivation, and the extracted lutein of the microalga to protect mammal cells against blue-light irradiation. The biomass productivity and lutein content of Chlorella sp. cultured in 20 g/L fertilizer medium for 6 days were 1.04 g/L/d and 4.41 mg/g, respectively. These values are approximately 1.3- and 1.4-fold higher than those achieved with the modified f/2 medium, respectively. The cost of medium per gram of microalgal biomass reduced by about 97%. The microalgal lutein content was further increased to 6.03 mg/g in 20 g/L fertilizer medium when supplemented with 20 mM urea, and the cost of medium per gram lutein reduced by about 96%. When doses of ≥1 μM microalgal lutein were used to protect mammal NIH/3T3 cells, there was a significant reduction in the levels of reactive oxygen species (ROS) produced by the cells in the following blue-light irradiation treatments. The results show that microalgal lutein produced by fertilizers with urea supplements has the potential to develop anti-blue-light oxidation products and reduce the economic challenges of microalgal biomass applied to carbon biofixation and biofuel production.

Agriculture of microalgae Chlorella vulgaris for polyunsaturated fatty acids (PUFAs) production employing palm oil mill effluents (POME) for future food, wastewater, and energy nexus

Malaysia is one of the second largest world palm oil exporters; correspondingly, the massive production of palm oil mill effluent (POME) is a significant concern. The present investigation aimed to isolate and characterize the potential microalgae and maximize the growth and biomass productivity of Chlorella sp. using POME. Chlorella vulgaris was cultured in BG-11 (control) and POME with 1:1, 1:2, 1:3, and 1:4 dilution ratios. Binary solvent extraction was applied to extract lipids, and polyunsaturated fatty acids (PUFAs) in the fatty acid methyl ethers were determined using gas chromatography coupled with mass spectrometry (GC-MS). Results suggest the lipid level of 21 wt.% was the highest amount obtained from biomass Chlorella vulgaris cultivated in 1:4 diluted POME. Chlorella sp. in POME with a 1:4 dilution ratio gives the highest biomass yield. It also yields the maximum dry cell weight of 0.42 g/L. Therefore, this study indicates that diluted POME with different dilutions is a suitable growth medium for Chlorella vulgaris biomass cultivation. Pairing wastewater with algae culture reduces biomass production and water treatment costs. Further studies demand more attention for future food, wastewater, and energy nexus.

Research on the manipulation of iron ions and alkalis in Chlorella vulgaris culture

• The experimental results shown that the variant concentration of iron ions in the medium and the variant pH of the initial algal fluid did different effects on the growth of Chlorella vulgaris . The Chlorella vulgaris can thrive better in the culture fluid with iron ions raising. The initial pH of the algae fluid affects its growth mainly in 1–3 days. Meanwhile, in a weak alkaline environment, such as pH 8.27 is beneficial to the growth of Chlorella vulgaris during the initial developing period. Also we find out in a certain range of alkaline environments such as pH 8 to 9 is the more conducive to its growth. By concluding the results of the research, we can obtain the optimal chlorella aquaculture production cultivation conditions with economic benefits. The use of microalgae as raw materials to produce various new biomass energy in the aquaculture industry has become a key issue in building a friendly ecology. However, maximizing algal growth and profit by regulating iron ion concentration (Fe 3+ ) and acid-base value (pH) remains a major challenge. Therefore, this study utilized Chlorella vulgaris to explore the influence of Fe 3+ concentration and initial pH value on algal growth to obtain optimum economic benefits and chemical culture conditions. Using an air-lift columnar photobioreactor for cultivation and a fluorescent lamp as a light source, the experiments were conducted at a light intensity of 3537 Lux at approximately 24 °C. To achieve economically optimal culture conditions, the daily OD 660 value of the Chlorella vulgaris cultures was used to explore the effects of different iron ion concentrations and initial pH values on algae growth and biomass accumulation. Our results demonstrated that different iron ion concentrations and pH conditions of the algal culture rendered different effects on Chlorella vulgaris growth. Overall, the enriched iron-containing medium was more favorable for algae growth. Over time, the medium with higher iron concentration (1.0 g/L) achieved higher OD faster and its accumulated chlorophyll was higher. Moreover, the initial pH of the algae culture medium affected its growth mainly within the first three days. Particularly, a weak alkaline environment (e.g., pH 8.27) benefited Chlorella vulgaris growth during the initial growth period. With the results of this study, our research determined the optimal Chlorella production conditions that maximize economic benefits.

Accurate assessment of the effect of Rhodamine 6G solution as a spectral converter on biomass production of microalgae Chlorella sp., nitrate uptake, and energy consumption by the light source.

Spectral conversion of light introduced as a new technique to manage incoming light photons into the culture medium of photosynthetic microorganisms which can increase biomass production. In this work, a much more accurate value of the effect of spectral conversion on the growth of Chlorella sp. was obtained. For this purpose, the light source and Rhodamine 6G solution, as a spectral converter, was placed in the center of the culture flask, which was covered with aluminum foil. The loss of photons was largely avoided due to the additional homogeneous distribution of light. Modification of white light using the solution has increased biomass productivity up to 300%. Also, for biomass productivity of 1mg L-1day-1, the energy consumption of the white light source was reduced up to 75%. Nitrate uptake has also increased up to 28%, and intracellular lipid content has decreased, as well.

Optimization of CO2 Supply for the Intensive Cultivation of Chlorella sorokiniana IPPAS C-1 in the Laboratory and Pilot-Scale Flat-Panel Photobioreactors.

Microalgae are increasingly being used for capturing carbon dioxide and converting it into valuable metabolites and biologically active compounds on an industrial scale. The efficient production of microalgae biomass requires the optimization of resources, including CO2. Here, we estimated the productivity of Chlorella sorokiniana IPPAS C-1 depending on CO2 concentrations and the ventilation coefficient of the gas-air mixture (GAM) in flat-panel photobioreactors (FP-PBRs) at laboratory (5 L) and pilot (18 L) scales. For the laboratory scale, the PBRs operated at 900 µmol quanta m−2 s−1 and 35.5 ± 0.5 °C; the optimal CO2 flow rate was estimated at 3 mL CO2 per 1 L of suspension per minute, which corresponds to 1.5% CO2 in the GAM and an aeration rate of 0.2 vvm. These parameters, being scaled up within the pilot PBRs, resulted in a high specific growth rate (µ ≈ 0.1 h−1) and high specific productivity (Psp ≈ 1 g dw L−1 d−1). The principles of increasing the efficiency of the intensive cultivation of C. sorokiniana IPPAS C-1 are discussed. These principles are relevant for the development of technological regimes for the industrial production of Chlorella in flat-panel PBRs of various sizes.

Biologically active or just “pseudo”-vitamin B12 as predominant form in algae-based nutritional supplements?

In order to find out whether algae-based nutritional supplements contain both physiologically active vitamin B12 and its non-active pseudo-form, a simple and fast ultra-high performance liquid chromatography method with UV detection was developed to separate these two forms. A total of 57 commercially available microalgae products based on Chlorella and Spirulina were analyzed. Results obtained showed a broad variety of the content of total vitamin B12 ranging from not-detectable to 445.9 μg 100g−1 dry matter for Chlorella samples, and between 92.8 and 164.1 μg 100g−1 dry matter for Spirulina products. However, there was a big variation in the concentration of pseudo-vitamin B12 within all samples analyzed. Chlorella products contained mainly physiologically active cobalamin, while pseudo-vitamin B12 was the prevailing form in Spirulina-labeled nutritional supplements. The importance of being able to differentiate between cobalamin and its pseudo-form in algae is not only relevant for food analysts, but also regarding consumer protection since they have to be able to rely on the correct labeling of nutritional supplements.

Biomass, pigment production, and nutrient uptake of Chlorella sp. under different photoperiods

Abstract. Fakhri M, Riyani E, Ekawati AW, Arifin NB, Yuniarti A, Widyawati Y, Saputra IK, Samuel PD, Arif MZ, Hariati AM. 2021. Biomass, pigment production, and nutrient uptake of Chlorella sp. under different photoperiods. Biodiversitas 22: 5344-5349. Chlorella sp. is well-known as a functional feed in fish culture and has been utilized in the food industry. In phototrophic cultivation, photoperiod plays a fundamental part in the growth and pigment content of microalgae. This work was purposed to evaluate the effect of light:dark cycle on the growth rate, production of biomass and pigment and nutrient utilization of Chlorella sp. Four photoperiods (8:16, 12:12, 18:6, and 24:0 h light:dark regimes) under a constant light intensity of 4500 lux were applied in this study. The results demonstrated that increasing light duration led to increased cell growth, biomass, and pigment production of Chlorella sp. The best cell concentration, specific growth rate, and biomass production were 28.5 x 106 cells mL-1, 1.47 day-1, and 0.815 g L-1 dry weight, respectively, under continuous illumination. The maximum chlorophyll a of 19.205 mg L-1 and carotenoid of 4.656 mg L-1 were obtained at 24:0 h photoperiod. The highest uptake of nitrate (66.331%) and phosphate (76.191%) by Chlorella sp. were achieved under 24 h light regime. Improving the uptake of nutrients resulted in enhanced growth and pigment content of Chlorella sp. We conclude that continuous illumination is the best photoperiod to produce biomass and pigment and improve the nutrient removal of Chlorella sp.

The utilization of photophosphorylation uncoupler to improve lipid production of Chlorella, a case study using transcriptome and functional gene expression analysis to reveal its mechanism

Uncoupling of photophosphorylation appears to have positive effect on lipid production of microalgae, however, underlying mechanisms involved in the effect are still unresolved. In this study, an uncoupler of photophosphorylation, carbonyl cyanide m-chlorophenyl hydrazone (CCCP), was used for the promotion of microalgal lipid accumulation. The results indicated that addition of CCCP was able to enhance the lipid productions of all tested species of Chlorella, especially when cultivated under mixotrophic condition. Based on the transcriptome and functional gene expression analyses, the functional genes involving in photosynthetic electron transport (PET), fatty acids synthesis, tricarboxylic acid (TCA) cycle, glycolysis and pentose phosphate (PP) pathway were found to be up-regulated while the genes participating in photosystem II, ATP synthesis during photosynthesis and calvin cycle were to be down-regulated by the presence of CCCP. According to these observations, the mechanism of CCCP to the promotion of lipid synthesis probably through channeling the NAD(P)H metabolic fluxes from carbon fixation towards fatty acid biosynthesis and lipid production, which was accomplished by (1) down-regulation of ribulose-1,5-bisphosphate carboxylase/oxygenase genes that involving in CO2 fixation via calvin cycle to reduce the NADPH consumption; (2) up-regulation of genes involved in TCA cycle and PP pathway to produce more NAD(P)H. The results of this study provide a new method to promote microalgal lipid production.

The growth, production and chemical compounds of Chlorella sp. in various concentrations of palm oil wastewater in laboratory culture media

The growth, production, and content of chemical compounds in Chlorella sp. in various concentrations of liquid waste from palm oil were examined from June to November 2019, located in the Palm Oil Industry, SeiGaluh, Kampar, Indonesia. Five different concentrations of oil palm wastewater, i.e., 15 %, 20 %, 25 %, 30 % and 0.0 % (control) were studied. The results of this study indicated that oil palm wastewater was able to increase the growth, production, and chemical content of Chlorella sp. The best growth for Chlorella sp. was found at a concentration of 25 % oil palm wastewater, with cell density (8.5 × 106 cells/mL), biomass (0.77 g/L), after being cultured for 13 days (culture period). Chemical compounds in Chlorella sp. also increased at a concentration of 25 %, and it was found that protein content was 45.09 %, lipid was 10.85 %, and carbohydrate was 12.77 %. The concentration of nitrate and phosphate in culture media was known to have decreased, where the highest reduction in nitrate occurred at the concentration of oil palm liquid waste of 25 %, from the initial state of 14.34 to 3.17 mg/L at the end of culture. There was a decrease in phosphate, from the initial conditions, 1.82 mg/L to 0.34. mg/L at the end of the study. Chlorella biomass harvested in this study was used as a complementary food material (pellets) for livestock in the province of Riau.

Evaluation of the operational conditions in the production and morphology of Chlorella sp.

It was evaluated the effect of operational conditions in the production of Chlorella sp. after its selection from genus Chlorella sp., Scenedesmus sp., Nannochloris sp., Tetraselmis sp. and Dunaliella salina. Microalgae were inoculated in drinking water with addition of NPK fertilizer (N 24%, P 24%, K 18%), at a concentration of 0.5 g/L, agitation of 150 rpm, temperature 25 °C, light intensity of 1680 lumens at a color temperature of 6400K, without pH control for 8 days. The cellular concentrations obtained were 3.72x107 (Chlorella sp.), 1.36x107 (Scenedesmus sp.), 3.55x107 (Tetraselmis sp.), 5.74x107 (Nannochloris sp.) and 3.45x106 (Dunaliella salina), where the microalgae Chlorella sp., shows invasive capacity in drinking water cultivations. Applying the 2n-p fractional factorial design concept for the elemental composition of the microalgae and the cellular morphology, it was obtained 44.33% of C, 7.09% of H, 8.53% of N and 0.84% ​​of S for the Chlorella sp.

Relative abundance of lipid types among Chlorella sp. and Scenedesmus sp. and ameliorating homogeneous acid catalytic conditions using central composite design (CCD) for maximizing fatty acid methyl ester yield

The present study has tested the biodiesel potential of two hyper lipid producing strains Chlorella sp. and Scenedesmus sp. in terms of biomass yield, quantity and quality of lipid and fatty acid composition. Biomass yield of Chlorella sp. and Scenedesmus sp. was 1.26 and 1.33 g/L, respectively on day 18 and 20. The lipid content and lipid productivity of Chlorella sp. and Scenedesmus sp. was estimated to be 21.3, 26.5% and 12.33, 14.74 mg/L/d, respectively. Notably, relative abundance of lipid types in both the strains revealed >60% neutral lipids followed by glycolipids and phospholipids in minimal level. Central composite design based optimization revealed 69 and 65.4% FAME yield from Chlorella sp. and Scenedesmus sp. at 3% sulphuric acid and 65 °C reaction temperature. Eventually, higher levels of saturated fatty acids (~45%) and monounsaturated fatty acids (~34%) and make Scenedesmus sp. a promising parent material for workable biodiesel production.

Adjustments of the photosynthetic unit and compensation mechanisms of tolerance to high ammonia concentration in Chlorella sp. grown in food waste digestate

Anaerobically digested food waste (ADF) is a waste effluent produced from anaerobic digestion of food wastes. It is difficult to treat but also holds high potential for valorization. Microalgae cultivation is a promising path for its treatment and valorization. However, ADF is very turbid and has toxic concentration of ammonia nitrogen (NH3-N) (up to 5000 mg L−1 NH3-N). Here we looked at the physiological responses and acclimation patterns of Chlorella sp., Mur 269 to toxicity and turbidity in ADF, compared to bolds basal medium (BBM), under different irradiances. Indicators obtained via pulse amplitude modulation (PAM) fluorometry such as electron transport rate (ETR), fv′/fm′ and alpha (α), alongside oxygenesis as an indicator of photosynthesis, were used to study the state of the photosystem of the organism. Pigment (chlorophylls and carotenoids) ratios were studied as indicators of adjustments in the photosynthetic unit and growth rates and productivities were used to monitor cell growth. μmax and biomass productivities were 63 and 47% higher in ADF than in BBM. μmax and biomass productivity of Chlorella sp., Mur 269, remained close to their optimal values at 0.681 ± 0.03 d−1 and 165 ± 8 mg L−1 d−1 even at high irradiance intensities of 1500 μmol m−2 s−1, indicating applicability of this species in outdoor cultures at high irradiance. On the other hand, the results showed that in comparison with BBM, the efficiency of Chlorella sp., Mur 269 photosystem II at optimum irradiance, as reflected in Fv′/Fm′ values, was reduced by 16% in ADF. A critical look at the photosynthesis of the species by PAM fluorometry revealed that the adaptive methods of this microalgae to toxicity involves adjustment of the photosynthetic unit to maximize absorption of light and compensation mechanisms for reductions in PSII activity including switching to mixotrophic growth mode.

Influence of zinc and manganese enrichments on growth, biosorption and photosynthetic efficiency of Chlorella sp.

Treating biosolids from industrial, urban, and agricultural plants produces high amounts of water. After organic pollutants and non-essential heavy metals have been removed, these wastewaters are still rich in trace elements such as zinc (Zn), copper, or manganese (Mn) and have high conductivity and extremely variable pH. In this study, an isolated Chlorella sp. strain was grown for 21days in nutrient solutions enriched with known amounts of Zn or Mn to obtain concentrations three (4.0mgL-1)- and six (1.0mgL-1)-fold higher than the basal medium levels, respectively, and over the limits permitted in aquatic environments. The green alga exhibited high tolerance to Zn and Mn, with the maximum abatement of Zn (28-30%) and Mn (60-63.5%) after 14 and 7days of culture, respectively. Mn stimulated the growth rate and biomass production of Chlorella, which showed the highest carbon levels just in the first week. In both treatments, the nitrogen and protein contents remarkably increased. The photosynthetic pigments increased until the 14th day, with a higher extent in the Zn-enriched solution. An increasing photochemical efficiency was observed after 7days of treatment, when the microalgae grown in Zn- and Mn-enriched solutions showed a slightly higher maximum photochemical efficiency than control. The autotrophic and controlled growth system adopted was designed to monitor the dynamic balance of Zn and Mn contents in the solutions and in the algal biomass. This system has proved to be useful in identifying the optimal nutritional conditions of the microalgae, along with the optimal temporal patterns of both metal biosorption capacity for water remediation and element bioaccumulation in the algal biomass.

Anaerobic digestion wastewater decolorization by H2O2-enhanced electro-Fenton coagulation following nutrients recovery via acid tolerant and protein-rich Chlorella production

Farmland wastewater such as swine waste effluent contains rich nutrients needed for the production of microalgae. However, the dark color and high turbidity in the effluent hinder light penetration and thus the photosynthesis of the microalgae. In this study, the H2O2-enhanced electro-Fenton coagulation (EFC) was employed to remove chemical oxygen demand (COD), total suspended solid (TSS), color, and phosphorus effectively in anaerobic digested (AD) swine wastewater. During the enhanced EFC process, lower pH (pH 3), lower temperature (10 °C), higher H2O2 concentration (>200 mg·L−1) and higher wastewater dilution factors (>2) all had positive effects on pollutant removal. After EFC pretreatment, the penetration of light into the medium was improved and the photosynthesis of Chlorella sp. was effectively increased. The initial N:P ratio in Chlorella cultures was adjusted to various levels (8:1 ~ 32:1) by KH2PO4 addition. When the N:P ratio was 8:1, the maximum biomass accumulation (2.07 g·L−1), maximum protein content (54.1%) and protein productivity (62.2 mg·L−1·d−1) were obtained. In the process of Chlorella growth, due to the rapid consumption of ammonia, the pH of wastewater reached as low as 4, but the Chlorella still survived, showing that this strain of Chlorella was strongly acid-tolerant. This study showed the H2O2-enhanced EFC as an effective means of pretreatment of anaerobic effluent for aqueous culture of protein-rich Chlorella, which demonstrated to be an effective combination for nutrients recycling, and favorable to high value-added biomass production.