A two-step sequential hydrothermal liquefaction (SEQHTL) model for simultaneous extraction of polysaccharide at the first step followed by bio-oil in the second was established. The effects of reaction temperature, residence time, and biomass/water ratio on the product distribution of each SEQHTL step were evaluated. Maximum yield (32 wt.%) of polysaccharides was obtained at 160 °C, 20 min and 1:9 biomass/water ratio. Considering the operation cost and bio-oil yield (>30%); 240 °C, 20 min and 1:9 biomass/water ratio was preferred as ideal SEQHTL condition for bio-oil extraction. SEQHTL always produced ∼5% more bio-oil and ∼50% less bio-char than direct hydrothermal liquefaction (DHTL). Free fatty acid content of the bio-oils exhibited a sharp decrease with increase in temperature. Comparative analysis of the energy input and net energy balance showed that SEQHTL requires ∼15% less MJ/kg bio-oil than DHTL. Energy recovery rate for SEQHTL is nearly 4% higher than the DHTL.
Hydrogen production from Arthrospira (Spirulina) platensis wet biomass through heterofermentation by the [FeFe] hydrogenase of hydrogenogens (hydrogen-producing bacteria) and autofermentation by the [NiFe] hydrogenase of Arthrospira platensis was discussed under dark anaerobic conditions. In heterofermentation, wet cyanobacterial biomass without pretreatment was hardly utilized by hydrogenogens for hydrogen production. But the carbohydrates in cyanobacterial cells released after cell wall disruption were effectively utilized by hydrogenogens for hydrogen production. Wet cyanobacterial biomass was pretreated with boiling and bead milling, ultrasonication, and ultrasonication and enzymatic hydrolysis. Wet cyanobacterial biomass pretreated with ultrasonication and enzymatic hydrolysis achieved the maximum reducing sugar yield of 0.407 g/g-DW (83.0% of the theoretical reducing sugar yield). Different concentrations (10 g/l to 40 g/l) of pretreated wet cyanobacterial biomass were used as substrate to produce fermentative hydrogen by hydrogenogens, which were domesticated with the pretreated wet cyanobacterial biomass as carbon source. The maximum hydrogen yield of 92.0 ml H2/g-DW was obtained at 20 g/l of wet cyanobacterial biomass. The main soluble metabolite products (SMPs) in the residual solutions from heterofermentation were acetate and butyrate. In autofermentation, hydrogen yield decreased from 51.4 ml H2/g-DW to 11.0 ml H2/g-DW with increasing substrate concentration from 1 g/l to 20 g/l. The main SMPs in the residual solutions from autofermentation were acetate and ethanol. The hydrogen production peak rate and hydrogen yield at 20 g/l of wet cyanobacterial biomass in heterofermentation showed 110- and 8.4-fold increases, respectively, relative to those in autofementation.
Astaxanthin (AXA) is a carotenoid with strong antioxidant activity that has been reported to have beneficial effects on human health. Unlike the case of Haematococcus pluvialis cysts, the study of other AXA-producing vegetative microalgae as alternative AXA sources has been minimal, because of inefficient AXA extraction yield and difficulty in separating chlorophyll and AXA. In this study, a novel method was developed to extract AXA from a vegetative green microalga (Monoraphidium sp. GK12) and to separate chlorophyll and AXA using a supercritical-CO2 extraction (SCE) and acid treatment. AXA in the microalgal biomass was extracted completely by SCE, while simple ethanol soaking could not extract AXA well. The addition of ethanol as a co-solvent improved AXA extraction yield. The method employed low CO2 pressure and reaction temperature as well as a short reaction time, making it energy-efficient. Chlorophyll in AXA-containing extract was easily removed by acid addition (H2SO4 or HCl) and centrifugation. Since those acids can be neutralized by alkaline, which produces nontoxic salt, and AXA in the extract can be condensed by reducing pressure, the developed method is expected to stimulate commercialization of a vegetative microalga as an alternative AXA source for food applications.
Optimization of the light conditions for biofuel production by the microalga Botryococcus braunii BOT-22 (race B) was performed using monochromatic red light. The lipid and sugar contents were approximately 40% and 20–30% of the cell dry weight, respectively, and about half of the lipids were liquid hydrocarbons. The half-saturation intensities for the production rate of lipids, hydrocarbons, and sugars were 63, 49, and 44 μmol m−2 s−1, respectively. Fluorescence microscopic images of Nile Red-stained cells showed an increased number of intracellular neutral lipid granules due to increased light intensity. After 16 days of incubation in the dark, lipid and sugar, but not hydrocarbon content decreased. Growth, metabolite production, and photosynthesis were saturated at 100, 200 and 1000 μmol m−2 s−1, respectively. These results indicate that photosynthetically captured energy is not used efficiently for metabolite production; thus, improvements in metabolic regulation may increase hydrocarbon production.
There is increasing interest in the use of microalgae as a renewable source for the production of fuels and chemicals, but improvements are needed in all steps of this process, including harvesting. A continuous microalgae harvest system was developed based on electrolysis, referred to here as a continuous electrolytic microalgae (CEM) harvest system. This innovative system combines cultivation and harvesting and enables continuous and efficient concentration of microalgae. The electrodes were subject to a polarity exchange (PE) in the middle of the operation to further improve the harvest efficiency. Use of PE, rather than conventional electro-coagulation-flotation (ECF), led to more efficient cell recovery and more uniform recovery over the entire harvest chamber. In addition, PE increased the cell growth rate and the circulated cells remained intact after harvesting.
There are two major energy and cost constraints to bulk production of single cell microalgae for biofuels or feed: expensive culture systems with high capital costs and high energy requirements for mixing and gas exchange; and the cost of harvesting using high-speed continuous centrifugation for dewatering. This report deals with the latter; harvesting by flocculation where theory states that alkaline flocculants neutralize the repelling surface charge of algal cells, allowing them to coalesce into a floc. It had been assumed that with such electrostatic flocculation, the more cells to be flocculated, the more flocculant needed, in a linear stoichiometric fashion, rendering flocculation overly expensive. Counter to theory of electrostatic flocculation, we find that the amount of alkaline flocculant needed is a function of the logarithm of cell density, with dense cultures requiring an order of magnitude less base than dilute suspensions, with flocculation occurring at a lower pH. Various other theories abound that flocculation can be due to multi-valent cross-linking, or co-precipitation with phosphate or with magnesium and calcium, but are clearly not relevant with the flocculants we used. Monovalent bases that cannot cross-link or precipitate phosphate work with the same log-linear stoichiometry as the divalent bases, obviating those theories, leaving electrostatic flocculation as the only tenable theory of flocculation with the materials used. The cost of flocculation of dense cultures with this procedure should be below $1.00/T algae for mixed calcium:magnesium hydroxides.
Microalgae exhibit a number of heavy metal uptake process by different metabolism. In this study, the ability of microalgae for removal of heavy metal from wastewater was studied. Growth and biochemical contents of microalgae were determined by spectrophotometer. Heavy metal analysis of wastewater effluents were performed by atomic absorption spectrophotometer before and after treatment at laboratory scale. The growth of Scenedesmus bijuga and Oscillatoria quadripunctulata in sewage wastewater was higher than those grown in synthetic medium. Whereas, the growth of S. bijuga and O. quadripunctulata in sterilized petrochemical effluents was slightly lower than that grown in the standard synthetic medium. The chlorophyll, carotenoid and protein content of S. bijuga and O. quadripunctulata grown in sterilized sewage wastewater were higher than those grown in the standard medium. Similarly S. bijuga and O. quadripunctulata grown in sterilized petrochemical effluents showed lower contents of pigments and protein than those grown in sewage and synthetic medium. Heavy metals copper, cobalt, lead and zinc were removed by 37-50, 20.3-33.3, 34.6-100 and 32.1-100%, respectively from sewage wastewater and petrochemical effluent using Ocillatoria culture. The metal absorption by S. bijuga were (Cu, Co, Pb, Zn) 60-50, 29.6-66, 15.4-25 and 42.9-50%, respectively from sewage and petrochemical effluents. Both species showed high level of heavy metal removal efficiency and metal sorption efficiency of both microalgae depended on the type of biosorbent, the physiological status of the cells, availability of heavy metal, concentration of heavy metal and chemical composition of wastewater.
Lipids from microalgae have become an important commodity in the last 20 years, biodiesel and supplementing human diets with ω-3 fatty acids are just two of the many applications. Acetyl-CoA carboxylase (ACCase) is a key enzyme in the lipid synthesis pathway. In general, ACCases consist of four functional domains: the biotin carboxylase (BC), the biotin carboxyl binding protein (BCCP), and α-and ß-carboxyltransferases (α-and ß-CT). In algae, like in plants, lipid synthesis is another function of the chloroplast. Despite being well researched in plants and animals, there is a distinct lack of information about this enzyme in the taxonomically diverse algae. In plastid-containing organisms, ACCases are present in the cytosol and the plastid (chloroplasts) and two different forms exist, the heteromeric (prokaryotic) and homomeric (eukaryotic) form. Despite recognition of the existence of the two ACCase forms, generalized published statements still list the heteromeric form as the one present in algal plastids. In this study, the authors show this is not the case for all algae. The presence of heteromeric or homomeric ACCase is dependent on the origin of plastid. The authors used ACCase amino acid sequence comparisons to show that green (Chlorophyta) and red (Rhodophyta) algae, with the exception of the green algal class Prasinophyceae, contain heteromeric ACCase in their plastids, which are of primary symbiotic origin and surrounded by two envelope membranes. In contrast, algal plastids surrounded by three to four membranes were derived through secondary endosymbiosis (Heterokontophyta and Haptophyta), as well as apicoplast containing Apicomplexa, contain homomeric ACCase in their plastids. Distinctive differences in the substrate binding regions of heteromeric and homomeric α-CT and β-CT were discovered, which can be used to distinguish between the two ACCase types. Furthermore, the acetyl-CoA binding region of homomeric α-CT can be used to distinguish between cytosolic and plastidial ACCase. The information provided here will be of fundamental importance in ACCase expression and activity research to unravel impacts of environmental and physicochemical parameters on lipid content and productivity.
Marine microalgae represent a potentially valuable feedstock for biofuel production; however, large-scale production is not yet economically viable. Optimisation of productivity and lipid yields is required and the cost of biomass harvesting and dewatering must be significantly reduced. Microalgae produce a wide variety of biologically active metabolites, many of which are involved in inter- and intra-specific interactions (the so-called infochemicals). The majority of infochemicals remain unidentified or uncharacterised. Here, we apply known and candidate (undefined extracts) infochemicals as a potential means to manipulate the growth and lipid content of Nannochloropsis oculata-a prospective species for biofuel production. Five known infochemicals (β-cyclocitral, trans,trans-2,4-decadienal, hydrogen peroxide, norharman and tryptamine) and crude extracts prepared from Skeletonema marinoi and Dunaliella salina cultures at different growth stages were assayed for impacts on N. oculata over 24 h. The neutral lipid content of N. oculata increased significantly with exposure to three infochemicals (β-cyclocitral, decadienal and norharman); however the effective concentrations affected a significant decrease in growth. Exposure to particular crude extracts significantly increased both growth and neutral lipid levels. In addition, water-soluble extracts of senescent S. marinoi cultures induced a degree of flocculation in the N. oculata. These preliminary results indicate that artificial manipulation of N. oculata cultures by application of algae infochemicals could provide a valuable tool towards achieving economically viable large-scale algae biofuel production.
Reduction of carbon loss from the effluent is one of the most important aspects of photobioreactors design. In this study, a novel gas sparger of bubble tank was adopted in a photobioreactor to enhance carbon dioxide (CO(2)) mass transfer rate as well as alleviate dissolved oxygen (DO) accumulation. The results showed that low DO level in the culture can be obtained due to the turbulent hydrodynamic condition provided by the bubble tank. The effects of CO(2) concentration, flow rate of influent, and light intensity on CO(2) removal efficiency were investigated. The maximum CO(2) removal efficiency was 94% at flow rate of 30mLmin(-1), light intensity of 179μmolm(-2)s(-1) and CO(2) concentration of 10%, implying that the novel gas sparger is a promising alternative for CO(2) removal from CO(2)-enriched air by cultivating microalgae in the photobioreactor.
Nitrogen-doped carbon materials are synthesized via an effective, sustainable, and green one-step route based on the hydrothermal carbonization of microalgae with high nitrogen content (ca. 11 wt %). The addition of the monosaccharide glucose to the reaction mixture is found to be advantageous, enhancing the fixation of nitrogen in the synthesized carbons, resulting in materials possessing nitrogen content in excess of 7 wt %, and leading to promising reaction yields. Increasing the amount of glucose leads to a higher nitrogen retention in the carbons, which suggests co-condensation of the microalgae and glucose-derived degradation/hydrolysis products via Maillard-type cascade reactions, yielding nitrogen-containing aromatic heterocycles (e.g., pyrroles) as confirmed by several analytical techniques. Increasing the HTC processing temperature leads to a further aromatization of the chemical structure of the HTC carbon and the formation of increasingly more condensed nitrogen-containing functional motifs (i.e., pyridinic and quaternary nitrogen).
The current interest in microalgae as a sustainable source of next generation biofuels and other valuable substances is driving exploration of their use as unique biotechnological production systems. To design and optimise appropriate production strategies, the behaviour of particular microalgal species should be well characterised under different culture conditions. Thus, flow cytometric (FCM) methods, which are already well established in environmental and toxicological studies of microalgae, are also useful for analysing the physiological state of microalgae, and have the potential to contribute to the rapid development of feasible bioprocesses. These methods are commonly based on the examination of intrinsic features of individual cells within a population (such as autofluorescence or size). Cells possessing the desired physiological or morphological features, which are detectable with or without fluorescent staining, are counted or isolated (sorted) using an FCM device. The options for implementation of FCM in the development of biotechnological processes detailed in this review are (i) analysing the chemical composition of biomass, (ii) monitoring cellular enzyme activity and cell viability, and (iii) sorting cells to isolate those overproducing the target compound or for the preparation of axenic cultures.
The feasibility of using a microalga Chlorella vulgaris YSW-04 was investigated for removal of nutrients from piggery wastewater effluent. The consequent lipid production by the microalga was also identified and quantitatively determined. The wastewater effluent was diluted to different concentrations ranging from 20 to 80 % of the original using either synthetic media or distilled water. The dilution effect on both lipid production and nutrient removal was evaluated, and growth rate of C. vulgaris was also monitored. Dilution of the wastewater effluent improved microalgal growth, lipid productivity, and nutrient removal. The growth rate of C. vulgaris was increased with decreased concentration of piggery wastewater in the culture media regardless of the diluent type. Lipid production was relatively higher when using synthetic media than using distilled water for dilution of wastewater. The composition of fatty acids accumulated in microalgal biomass was dependent upon both dilution ratio and diluent type. The microalga grown on a 20 % concentration of wastewater effluent diluted with distilled water was more promising for generating high-efficient biodiesel compared to the other culture conditions. The highest removal of inorganic nutrients was also achieved at the same dilution condition. Our results revealed the optimal pretreatment condition for the biodegradation of piggery wastewater with microalgae for subsequent production of high-efficient biodiesel.
Factors such as an aging population, increased health care costs and rapid advances in science and technology are likely driving the increase interest among consumers in attaining wellness through diet, which is in turn, fuelling interest in functional foods and changing the way that people eat.
Microalgae have been largely cultured and commercialized as food and feed additives, their potential as source of high-added value compounds and their ability to positively affect human’s health due to their original chemical composition, is well known. Considering pasta is a main staple food, the objective of this study was to prepare fresh spaghetti enriched with different amounts of microalgae and to compare the fatty acid profile of pastas before and after cooking, with standard semolina spaghetti.
The results show that fatty acid profile of pastas prepared with Isochrysis galbana and Diacronema vlkianum biomass incorporation, presented a high resistance to the thermal treatment applied during the cooking procedure. The increase of the amount of the algae lead to a significant increase of EPA (Eicosapentaenoic Acid) and DHA (Docosahexaenoic Acid) both in raw and cooked pastas, omega-3 fatty acids that can be obtained through seafood.
This paper primarily presents an overall review of the use of microalgae as a biofuel feedstock. Among the microalgae that have potential as biofuel feedstock, Chlorella, specifically, was thoroughly discussed because of its ability to adapt both to heterotrophic and phototrophic culture conditions. The lipid content and biomass productivity of microalgae can be up to 80% and 7.3 g/l/d based on the dried weight of biomass, respectively, making microalgae an ideal candidate as a biofuel feedstock. The set-up of the system and the biomass productivity of microalgae cultivated in an open pond and a photobioreactor were also compared in this work. The effect of the culture condition is discussed based on the two-stage culture period. The issues that were discussed include the light condition and the CO2, DO and N supply. The microalgal productivities under heterotrophic and phototrophic culture conditions were also compared and highlighted in this work. The harvesting process and type of flocculants used to aid the harvesting were highlighted by considering the final yield of biomass. A new idea regarding how to harvest microalgae based on positive and negative charges was also proposed in this work. The extraction methods and solvents discussed were primarily for the conventional and newly invented techniques. Conversion processes such as transesterification and thermochemical processes were discussed, sketched in figures and summarized in tables. The cost–benefit analysis of heterotrophic culture and the cultivation system was highlighted at the end of this work. Other benefits of microalgae are also mentioned in this work to give further support for the use of microalgae as a feedstock for biofuel production.
In order to utilize microalgae and their residues as biomass solid fuels for co-firing in the existing pulverized coal-fired power plants, this study is to investigate the properties of fuels from those feedstock prepared by the torrefaction process. The microalgae were cultivated by using flue gas emitted from coal-fired power plants of Taipower Co., and the residue was from extraction of microalgae by hot water. The operating conditions for the torrefaction temperature and residence time were set as 200, 250, 300 and 350 °C, and 30, 60, 120 min, respectively. Also, 30 °C/min and 50 °C/min as the heating rates were chosen to examine the mass yield of solid torrefied biomass from microalgae. After torrefaction, the ultimate analysis, proximate analysis, and higher heating value analysis of the torrefied biomass were carried out. In addition, the Hardgrove Grindability Index (HGI) of microalgal residue and its torrefied solid were measured. The results show that carbon content, ash content, fixed carbon content, higher heating value, and HGI of torrefied biomass increased with increasing the operating temperature and residence time. At the condition of 300 °C and 30 min, the higher heating value of torrefied microalgae and microalgal residue were 25.92 MJ/kg and 26.76 MJ/kg, respectively. Compared to raw materials, the increases were 26.70% and 17.16%, respectively. At the same circumstances, hydrogen content, oxygen content, moisture content and mass yield of solid torrefied biomass decreased with increasing temperature and residence time. After torrefaction at 300 °C, the moisture content were all down to 1%, and the mass yield of solid torrefied biomass under 350 °C by torrefaction were also kept up to 50%. The HGI of torrefied microalgal residue was 48.53, which exceed the sub-bituminous coal when the operation condition temperature was up to 250 °C.
The use flocculants on the recovery of freshwater microalgae is studied. Flocculants tested include metal salts, chitosan, and polyelectrolytes used in wastewater treatment processes. Influence of flocculant, but also the doses and biomass concentrations affecting biomass recovery as well as the concentration factor has been evaluated. Results showed that the use of metal salts or chitosan was not efficient, whereas polyelectrolytes allow the efficient recovery of biomass, at doses of 2-25 mg per gram of microalgae biomass. The required doses depend on the particular polyelectrolyte and the freshwater strain present; but cationic polyelectrolytes are generally recommended. The use of polyelectrolytes does not adversely affect water reuse in the production process. The concentration factors obtained are higher than 35 in most cases. Such high concentration factors allow a reduction in the equipment size necessary for biomass dewatering, thus improving the viability of using these microorganisms in biofuel or wastewater processes.
In this study, the focus is on magnetic separation of fresh water algae Chlamydomonas reinhardtii and Chlorella vulgaris as well as marine algae Phaeodactylum tricornutum and Nannochloropsis salina by means of silica-coated magnetic particles. Due to their small size and low biomass concentrations, harvesting algae by conventional methods is often inefficient and cost-consuming. Magnetic separation is a powerful tool to capture algae by adsorption to submicron-sized magnetic particles. Hereby, separation efficiency depends on parameters such as particle concentration, pH and medium composition. Separation efficiencies of > 95 % were obtained for all algae while maximum particle loads of 30 g/g and 77 g/g were measured for C. reinhardtii and P. tricornutum at pH 8 and 12, respectively. This study highlights the potential of silica-coated magnetic particles for the removal of fresh water and marine algae by high gradient magnetic filtration and provides critical discussion on future improvements.
Excess greenhouse gas emissions and the concomitant effect on global warming have become significant environmental, social and economic threats. In this context, the development of renewable, carbon-neutral and economically feasible biofuels is a driving force for innovation worldwide. A lot of effort has been put into developing biodiesel from microalgae. However, there are still a number of technological, market and policy barriers that are serious obstacles to the economic feasibility and competitiveness of such biofuels. Conversely, there are also a number of business opportunities if the production of such alternative biofuel becomes part of a larger integrated system following the Biorefinery strategy. In this case, other biofuels and chemical products of high added value are produced, contributing to an overall enhancement of the economic viability of the whole integrated system. Additionally, dual purpose microalgae-bacteria-based systems for treating wastewater and production of biofuels and chemical products significantly contribute to a substantial saving in the overall cost of microalgae biomass production. These types of systems could help to improve the competitiveness of biodiesel production from microalgae, according to some recent Life Cycle Analysis studies. Furthermore, they do not compete for fresh water resources for agricultural purposes and add value to treating the wastewater itself. This work reviews the most recent and relevant information about these types of dual purpose systems. Several aspects related to the treatment of municipal and animal wastewater with simultaneous recovery of microalgae with potential for biodiesel production are discussed. The use of pre-treated waste or anaerobic effluents from digested waste as nutrient additives for weak wastewater is reviewed. Isolation and screening of microalgae/cyanobacteria or their consortia from various wastewater streams, and studies related to population dynamics in mixed cultures, are highlighted as very relevant fields of research. The species selection may depend on various factors, such as the biomass and lipid productivity of each strain, the characteristics of the wastewater, the original habitat of the strain and the climatic conditions in the treatment plant, among others. Some alternative technologies aimed at harvesting biomass at a low cost, such as cell immobilization, biofilm formation, flocculation and bio-flocculation, are also reviewed. Finally, a Biorefinery design is presented that integrates the treatment of municipal wastewater with the recovery of oleaginous microalgae, together with the use of seawater supplemented with anaerobically digested piggery waste for cultivating Arthrospira (Spirulina) and producing biogas, biodiesel, hydrogen and other high added value products. Such strategies offer new opportunities for the cost-effective and competitive production of biofuels along with valuable non-fuel products.
Chlorella ellipsoidea is a single-celled eukaryotic green microalgae with high nutritional value. Its value may be further increased if a simple, reliable and cost-effective transformation method for C. ellipsoidea can be developed. In this paper, we describe a novel transformation method for C. ellipsoidea . This system is based on treatment of C. ellipsoidea cells with cellulolytic enzymes to weaken their cell walls, making them become competent to take up foreign DNA. To demonstrate the usefulness and effectiveness of this method, we treated C. ellipsoidea cells with a cell wall-degrading enzyme, cellulase, followed by transformation with plasmid pSP-Ubi-GUS harbouring both the zeocin resistance gene and the beta-glucuronidase (GUS) reporter gene that serve as selective makers for transformation. Transformants were readily obtained on zeocin selection medium, reaching transformation efficiency of 2.25 × 10(3) transformants/μg of plasmid DNA. PCR analysis has also demonstrated the presence of the GUS reporter gene in the zeocin-resistant transformants. Histochemical assays further showed the expression of the GUS activity in both primary transformants and transformants after long-term growth (10 months) with antibiotic selection on and off. Availability of a simple and efficient transformation system for C. ellipsoidea will accelerate the exploration of this microalga for a broader range of biotechnological applications, including its use as a biologic factory for the production of high-value human therapeutic proteins