Metabolomic Study of Heterotrophically Grown Chlorella sp. Isolated from Wastewater in Northern Sweden

There are numerous strains of Chlorella with a corresponding variety of metabolic pathways. A strain we previously isolated from wastewater in northern Sweden can grow heterotrophically as well as autotrophically in light and has higher lipid contents under heterotrophic growth conditions. The aims of the present study were to characterize metabolic changes associated with the higher lipid contents in order to enhance our understanding of lipid production in microalgae and potentially identify new compounds with utility in sustainable development. Inter alia, the amino acids glutamine and lysine were 7-fold more abundant under heterotrophic conditions, the key metabolic intermediate alpha-ketoglutarate was more abundant under heterotrophic conditions with glucose, and maltose was more abundant under heterotrophic conditions with glycerol than under autotrophic conditions. The metabolite 3-hydroxy-butyric acid, the direct precursor of the biodegradable plastic PHB (poly-3-hydroxy-butyric acid), was also more abundant under heterotrophic conditions. Our metabolomic analysis has provided new insights into the alga’s lipid production pathways and identified metabolites with potential use in sustainable development, such as the production of renewable, biodegradable plastics, cosmetics, and nutraceuticals, with reduced pollution and improvements in both ecological and human health.     

Cultivation of Chlorella vulgaris in Dairy Wastewater Pretreated by UV Irradiation and Sodium Hypochlorite

There is potential in the utilization of microalgae for the purification of wastewater as well as recycling the resource in the wastewater to produce biodiesel. The large-scale cultivation of microalgae requires pretreatment of the wastewater to eliminate bacteria and protozoa. This procedure is costly and complex. In this study, two methods of pretreatment, UV irradiation, and sodium hypochlorite (NaClO), in various doses and concentrations, were tested in the dairy wastewater. Combining the efficiency of biodiesel production, we proposed to treat the dairy wastewater with NaClO in the concentration of 30 ppm. In this condition, The highest biomass productivity and lipid productivity of Chlorella vulgaris reached 0.450 g L^?1 day^?1 and 51 mg L^?1 day^?1 after a 4-day cultivation in the dairy wastewater, respectively.     

Cultivation of Chlorella protothecoides with Urban Wastewater in Continuous Photobioreactor: Biomass Productivity and Nutrient Removal

The capability to grow microalgae in nonsterilized wastewater is essential for an application of this technology in an actual industrial process. Batch experiments were carried out with the species in nonsterilized urban wastewater from local treatment plants to measure both the algal growth and the nutrient consumption. Chlorella protothecoides showed a high specific growth rate (about 1 day^?1), and no effects of bacterial contamination were observed. Then, this microalgae was grown in a continuous photobioreactor with CO₂–air aeration in order to verify the feasibility of an integrated process of the removal of nutrient from real wastewaters. Different residence times were tested, and biomass productivity and nutrients removal were measured. A maximum of microalgae productivity was found at around 0.8 day of residence time in agreement with theoretical expectation in the case of light-limited cultures. In addition, N-NH₄ and P-PO₄ removal rates were determined in order to model the kinetic of nutrients uptake. Results from batch and continuous experiments were used to propose an integrated process scheme of wastewater treatment at industrial scale including a section with C. protothecoides.     

Lipid Production by Culturing Oleaginous Yeast and Algae with Food Waste and Municipal Wastewater in an Integrated Process

Food waste and municipal wastewater are promising feedstocks for microbial lipid biofuel production, and corresponding production process is to be developed. In this study, different oleaginous yeast strains were tested to grow in hydrolyzed food waste, and growths of Cryptococcus curvatus, Yarrowia lipolytica, and Rhodotorula glutinis in this condition were at same level as in glucose culture as control. These strains were further tested to grow in municipal primary wastewater. C. curvatus and R. glutinis had higher production than Y. lipolytica in media made from primary wastewater, both with and without glucose supplemented. Finally, a process was tested to grow C. curvatus and R. glutinis in media made from food waste and municipal wastewater, and the effluents from these processes were further treated with yeast culture and phototrophic algae culture; 1.1 g/L C. curvatus and 1.5 g/L R. glutinis biomass were further produced in second-step yeast cultures, as well as 1.53 and 0.58 g/L Chlorella sorokiniana biomass in phototrophic cultures. The residual nitrogen concentrations in final effluents were 33 mg/L and 34 mg/L, respectively, and the residual phosphorus concentrations were 1.5 and 0.6 mg/L, respectively. The lipid contents in the produced biomass were from 18.7% to 28.6%.     

Marine Diatom, Navicula sp. Strain JPCC DA0580 and Marine Green Alga, Chlorella sp. Strain NKG400014 as Potential Sources for Biodiesel Production

Marine diatom, strain JPCC DA0580, and marine green microalga strain NKG400014 were selected as high neutral lipid-producers from marine microalgal culture collection toward biodiesel production. These strains were tentatively identified as Navicula sp. and Chlorella sp., respectively, by 18S rDNA analysis. Growth and lipid accumulation conditions of both strains were analyzed by changing nutrient concentrations in growth media and initial illuminance intensity. The highest productivity of fatty acid methyl ester (FAME) reached to 154 mg/L/week for NKG400014 and 185 mg/L/week for JPCC DA0580. Gas chromatography/mass spectrometry analysis indicates that FAME fraction from NKG400014 mainly contained 9-12-15-octadecatrienoate (C18:3) and that from JPCC DA0580 mainly contained methyl palmitate (C16:0) and methyl palmitoleate (C16:1). Furthermore, calorimetric analysis revealed that the energy content of strain was 4,233?±?55 kcal/kg (i.e., 15.9?±?0.2 MJ/kg) for NKG400014 and 6,423?±?139 kcal/mg (i.e., 26.9?±?0.6 MJ/kg) for JPCC DA0580, respectively. The value from JPCC DA0580 was equivalent to that of coal. The strains NKG400014 and JPCC DA0580 will become a promising resource that can grow as dominant species in the open ocean toward production of both liquid and solid biofuels.     

Growth and Nutrient Utilization of Green Algae in Batch and Semicontinuous Autotrophic Cultivation Under High CO2 Concentration

Applied Biochemistry and Biotechnology - The growth performance of Chlorella protothecoides, Chlorella pyrenoidosa, and Chlorella sp. in autotrophic cultivation with 10% carbon dioxide (CO2) was...     

城市污水处理厂污泥中卤代甲基磺酸的液相色谱-串联质谱分析方法研究

建立了测定城市污水处理厂污泥样品中5种卤代甲基磺酸的高效液相色谱-串联质谱(HPLC-MS/MS)方法。通过对萃取溶剂、提取方式和萃取次数等条件进行优化,确定最佳萃取条件为:萃取溶剂为甲醇-水(体积比1∶1)混合溶液,摇床振荡提取1 h,萃取1次。在该条件下对一氯甲基磺酸(Cl-MSA)、二氯甲基磺酸(Cl_2-MSA)、三氯甲基磺酸(Cl_3-MSA)、一溴甲基磺酸(Br-MSA)和三氟甲基磺酸(F_3-MSA)的萃取效率分别为(48. 4±2. 8)%、(68. 3±0. 9)%、(81. 0±4. 8)%、(38. 0±1. 1)%和(99. 1±1. 2)%。该方法的线性范围为0. 05~50μg·kg~(-1),相关系数(r)大于0. 991,各化合物的检出限(S/N=3)为0. 003~0. 018μg·kg~(-1),定量下限为0. 010~0. 059μg·kg~(-1);对实际样品的加标回收率为73. 8%~101%,相对标准偏差为2. 3%~12. 5%,可满足污泥样品中痕量卤代甲基磺酸的分析需求。     

Pretreatment for Simultaneous Production of Total Lipids and Fermentable Sugars from Marine Alga, Chlorella sp.

The goal of this study was to determine the optimal pretreatment process for the extraction of lipids and reducing sugars to facilitate the simultaneous production of biodiesel and bioethanol from the marine microalga Chorella sp. With a single pretreatment process, the optimal ultrasonication pretreatment process was 10 min at 47 KHz, and extraction yields of 6.5 and 7.1 (percentage, w/w) of the lipids and reducing sugars, respectively, were obtained. The optimal microwave pretreatment process was 10 min at 2,450 MHz, and extraction yields of 6.6 and 7.0 (percentage, w/w) of the lipids and reducing sugars, respectively, were obtained. Lastly, the optimal high-pressure homogenization pretreatment process was two cycles at a pressure of 20,000 psi, and extraction yields of 12.5 and 12.8 (percentage, w/w) of the lipids and reducing sugars, respectively, were obtained. However, because the single pretreatment processes did not markedly improve the extraction yields compared to the results of previous studies, a combination of two pretreatment processes was applied. The yields of lipids and reducing sugars from the combined application of the high-pressure homogenization process and the microwave process were 24.4 and 24.9 % (w/w), respectively, which was up to three times greater than the yields obtained using the single pretreatment processes. Furthermore, the oleic acid content, which is a fatty acid suitable for biodiesel production, was 23.39 % of the fatty acids (w/w). The contents of glucose and xylose, which are among the fermentable sugars useful for bioethanol production, were 77.5 and 13.3 % (w/w) of the fermentable sugars, respectively, suggesting the possibility of simultaneously producing biodiesel and bioethanol. Based on the results of this study, the combined application of the high-pressure homogenization and microwave pretreatment processes is the optimal method to increase the extraction yields of lipids and reducing sugars that are essential for the simultaneous production of biodiesel and bioethanol.     

Kinetic Model of Photoautotrophic Growth of Chlorella sp. Microalga,Isolated from the Setúbal Lagoon

In this work, a kinetic expression relating light availability in the culture medium with the rate of microalgal growth is obtained. This expression, which is valid for low illumination conditions, was derived from the reactions that take part in the light‐dependent stage of photosynthesis. The kinetic expression obtained is a function of the biomass concentration in the culture, as well as of the local volumetric rate of absorption of photons, and only includes two adjustable parameters. To determine the value of these parameters and to test the validity of the hypotheses made, autotrophic cultures of the Chlorella sp. strain were carried out in a modified BBM medium at three CO₂ concentrations in the gas stream, namely 0.034%, 0.34% and 3.4%. Moreover, the local volumetric rate of photon absorption was predicted based on a physical model of the interaction of the radiant energy with the suspended biomass, together with a Monte Carlo simulation algorithm. The proposed intrinsic expression of the biomass growth rate, together with the Monte Carlo radiation field simulator, are key to scale up photobioreactors when operating under low irradiation conditions, independently of the configuration of the reactor and of its light source.     

Chemical Characterization,Antiproliferative and Antioxidant Activities of Polyunsaturated Fatty Acid-Rich Extracts from Chlorella sp. S14

Microalgae is a rich source of polyunsaturated fatty acid. This study was conducted to identify and isolate microalgal strain with the potentials for producing polyunsaturated fatty acids (PUFAs) and determine its cytotoxic effect on some cancer cells. The algal strain (Chlorella sp. S14) was cultivated using modified BG-11 media, and algal biomass obtained was used for fatty acid extraction. Gas chromatographic–mass spectrometry was used to identify and quantify the levels of the fatty acid constituents. The total content of monounsaturated fatty acids (1.12%) was low compared to polyunsaturated fatty acids (PUFAs) (52.87%). Furthermore, n-3 PUFAs accounted for (12.37%) of total PUFAs with the presence of α-linolenic acid (2.16%) and cis-11,14,17-eicosatrienoic acid (2.16%). The PUFA-rich extract did not exhibit a cytotoxic effect on normal cells. Treatment with the PUFA-rich extract (150 µg/mL) significantly reduced cell viability in MCF-7 (31.58%) and A549 (62.56%) cells after the 48 h treatment. Furthermore, treatment of MCF-7 with fatty acid extracts (125 and 150 µg/mL) showed a significant reduction in MDA levels, increase in catalase activities and decrease in GSH level compared to untreated cells. However, a slight decrease in MDA level was observed in A549 cells after the 48 h treatment. There are no significant changes in catalase activities and GSH level in treated A549 cells. However, a slight reduction of NO levels was observed in treated MCF-7 and A549 cells. These results indicate the potentials of PUFA-rich extracts from Chlorella sp. S14 to reduce viability and modulate redox status in A549 and MCF-7 cells.     

Enhanced Crude Oil Sorption by Modified Plant Materials in Oilfield Wastewater Treatment

The treatment of oilfield wastewater with high crude oil content and complex composition is a problem requiring considerable attention. In order to effectively remove crude oil contained in wastewater, in this work, rice straw, as an oil-absorbing material, was modified and used as a sorbent for crude oil. Rice straw was modified with alkali and cetyltrimethylammonium chloride (CTAC) by simple substitution reaction. The adsorption capacity of modified rice straw for oil was evaluated. The results illustrate that the adsorption rate of rice straw for crude oil was increased from 0.83 to 8.49 g/g, with the optimal conditions of 18% NaOH reacted for 90 min at 50 °C and 2% CTAC reacted for 60 min at 20 °C. The proposed modification method could be used for different materials to enhance the adsorption rate. The results of the contact angle test show that the modified straw changed from hydrophilic to hydrophobic, which may be the main reason for the improvement in the oil absorption rate. Finally, the surface structure of rice straw was characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR) and N₂ adsorption–desorption isotherms, which further confirmed the hydrophobicity of the modified rice straw.     

Effect of Microalgae/Activated Sludge Ratio on Cooperative Treatment of Anaerobic Effluent of Municipal Wastewater

In this work, capability of the green microalga (MA), Chlorella vulgaris, in treating synthetic anaerobic effluent of municipal wastewater was investigated. While pure C. vulgaris (100 % MA) provided maximum soluble chemical oxygen demand (sCOD) and N???NH ₄ ⁺ removal efficiencies of 27 and 72 % respectively, addition of activated sludge (AS) to MA in different mass ratios (91, 80, 66.7, 9 % MA) improved wastewater treatment efficiency. Thus giving maximum sCOD and N???NH ₄ ⁺ removal efficiencies 85 and 86.3 % (for MA/AS?=?10/1), respectively. Utilizing AS without C. vulgaris, for treating the synthetic wastewater resulted in 87 % maximum sCOD and 42 % maximum N???NH ₄ ⁺ removal efficiencies. Furthermore, algal growth and specific growth rates were measured in the systems with microalga as the dominant cellular population. As a result, faster algal growth was observed in mixed systems. Specific growth rate of C. vulgaris was 0.14 (day^?1) in 100 % MA and 0.39 (day^?1) in 80 % MA. Finally, data gathered by online measurement of dissolved oxygen indicate that algae-activated sludge mixture improves photosynthetic activity of examined microalga strain during anaerobic effluent treatment.     

Literally Green Chemical Synthesis of Artemisinin from Plant Extracts

Active pharmaceutical ingredients are either extracted from biological sources—where they are synthesized in complex, dynamic environments—or prepared in stepwise chemical syntheses by reacting pure reagents and catalysts under controlled conditions. A combination of these two approaches, where plant extracts containing reagents and catalysts are utilized in intensified chemical syntheses, creates expedient and sustainable processes. We illustrate this principle by reacting crude plant extract, oxygen, acid, and light to produce artemisinin, a key active pharmaceutical ingredient of the most powerful antimalarial drugs. The traditionally discarded extract of Artemisia annua plants contains dihydroartemisinic acid—the final biosynthetic precursor—as well as chlorophyll, which acts as a photosensitizer. Efficient irradiation with visible light in a continuous‐flow setup produces artemisinin in high yield, and the artificial biosynthetic process outperforms syntheses with pure reagents.     

Effectiveness of Sal Deoiled Seed Cake as an Inducer for Protease Production from Aeromonas sp. S1 for its Application in Kitchen Wastewater Treatment

The present study is an attempt to demonstrate the feasibility of sal (Shorea robusta) deoiled cake—a forest-based industrial by-product—as a cheaper media supplement for augmented protease production from Aeromonas sp. S1 and application of protease in the treatment of kitchen wastewater. Under optimized conditions, protease production could successfully be enhanced to 5.13-fold (527.5 U mL^?1) on using sal deoiled seed cake extract (SDOCE), as medium additive, compared to an initial production of 102.7 U mL^?1 in its absence. The culture parameters for optimum production of protease were determined to be incubation time (48 h), pH (7.0), SDOCE concentration (3 % (v/v)), inoculum size (0.3–0.6 % (v/v)), and agitation rate (100 rpm). The enzyme was found to have an optimum pH and temperature of 8.0 and 60 °C, respectively. The protease preparation was tested for treatment of organic-laden kitchen wastewater. After 96 h of wastewater treatment under static condition, enzyme preparation was able to reduce 74 % biological oxygen demand, 37 % total suspended solids, and 41 % oil and grease. The higher and improved level of protease obtained using sal deoiled seed cake-based media hence offers a new approach for value addition to this underutilized biomass through industrial enzyme production. The protease produced using this biomass could also be used as pretreatment tool for remediation of organic-rich food wastewater.     

红树植物海杧果内生真菌Penicillium sp.中的抗菌活性成分

对红树植物海杧果内生真菌Penicillium sp.发酵液的乙酸乙酯提取物进行了柱层析分离, 得到3个化合物, 经MS, NMR, 1H-1H COSY, HMQC和HMBC等鉴定, 分别为4-(3-hydroxybutan-2-yl)-3,6-dimethylbenzene-1,2-diol(1), 4-(3-hydroxybutan-2-yl)-3-methyl-6-acetylbenzene-1,2-diol(2)和3,4,5-trimethyl-1,2-benzenediol(3). 其中化合物1和2为新化合物, 化合物3为新的天然产物. 抗菌活性测试结果表明, 化合物1和3均具有抗耐甲氧西林金黄色葡萄球菌的活性, 化合物2则未显示此活性.     

模拟胃肠道消化液中小球藻类金属硫蛋白对镉离子的结合作用研究

利用锌诱导小球藻,通过提取分离和纯化,获得锌结合类金属硫蛋白(MT-like蛋白).通过透析过程,研究模拟胃、肠道消化液中MT-like蛋白对Cd2+的结合作用.结果表明,在模拟胃液中MT-like蛋白处于脱金属状态,与Cd2+的结合能力弱.而在肠消化液中,MT-like蛋白能与Cd2+结合,20 mg·L-1 MT-...     

核事故产生的放射性废水处理方法研究进展

核事故会对人们生命安全及环境健康造成巨大危害。为最大限度地控制放射性物质的传播扩散,放射性废水的有效处理和处置应当引起我们足够的重视。本文总结了切尔诺贝利核爆炸、日本福岛核泄漏对放射性废水的处理经验,列举了美国、英国等国推荐使用的有效处理方法,并对吸附法、离子交换法、膜分离法以及联用工艺的研究进展进行了概述,为我国建立和完善核事故放射性废水处理和处置方案提供借鉴。     

解偶联剂CCCP对莱茵衣藻光照产氢过程的调控

本文研究莱茵衣藻在含硫和缺硫连续光照条件下, 不同浓度的CCCP对PSⅡ光化学效率、 光照产氢、 光合放氧以及光能的吸收和转换效率等的影响和调控规律.     

LC Determination of 4-Bromoaniline in Green Algae Chlamydomonas reinhardtii After Continuous-Flow Microextraction

In this study, the determination of 4-Bromoaniline (4-BA) in green algae Chlamydomonas reinhardtii (C. reinhardtii) was investigated by applying continuous-flow microextraction (CFME) combined with high-performance liquid chromatography (HPLC). Continuous-flow microextraction was conducted in a homemade glass chamber, i.e. the sample solution flowed through a constant volume drop of solvent in the chamber at a constant flow rate. The effects of different factors on extraction efficiencies were also investigated and these factors included the kind of extraction solvent, solvent drop volume, sample flow rate, extraction time and addition amount of salt. Under the optimum extraction conditions (extraction solvent, carbon tetrachloride; solvent drop volume, 3.5 μL; sample flow rate, 1.0 mL min⁻¹; extraction time, 10 min; no addition of salt), the calibration plot was set up by plotting peak area against a series of 4-Bromoaniline concentrations (0.01–10 μg mL⁻¹) in aqueous solution. The correlation coefficient (r) was 0.9990. The limit of detection (LOD) was 0.6 ng mL⁻¹. The precision of this method was obtained by successive five time analyses of 100-ng mL⁻¹ standard solution of 4-Bromoaniline, and the relative standard deviation (RSD) was 3.5%. The concentration factor was calculated by the ratio of peak area of the analyte obtained after and before extraction and found to be 10.6. 4-Bromoaniline residues in Chlamydomonas. reinhardtii cells and tap water samples were satisfactorily analyzed according to the method described above.