Strategies to enhance production of microalgal biomass and lipids for biofuel feedstock

ABSTRACT

Microalgae have enormous potential as feedstock for biofuel production compared with other sources, due to their high areal productivity, relatively low environmental impact, and low impact on food security. However, high production costs are the major limitation for commercialization of algal biofuels. Strategies to maximize biomass and lipid production are crucial for improving the economics of using microalgae for biofuels. Selection of suitable algal strains, preferably from indigenous habitats, and further improvement of those ‘platform strains’ using mutagenesis and genetic engineering approaches are desirable. Conventional approaches to improve biomass and lipid productivity of microalgae mainly involve manipulation of nutritional (e.g. nitrogen and phosphorus) and environmental (e.g. temperature, light and salinity) factors. Approaches such as the addition of phytohormones, genetic and metabolic engineering, and co-cultivation of microalgae with yeasts and bacteria are more recent strategies to enhance biomass and lipid productivity of microalgae. Improvement in culture systems and the use of a hybrid system (i.e. a combination of open ponds and photobioreactors) is another strategy to optimize algal biomass and lipid production. In addition, the use of low-cost substrates such as agri-industrial wastewater for the cultivation of microalgae will be a smart strategy to reduce production costs. Such systems not only generate high algal biomass and lipid productivity, but are also useful for bioremediation of wastewater and bioremoval of waste CO₂. The aim of this review is to highlight the advances in the use of various strategies to enhance production of algal biomass and lipids for biofuel feedstock.     

Photoautotrophic cultivating options of freshwater green microalgal Chlorococcum humicola for biomass and carotenoid production

Photoautotrophic cultivation of Chlorococcum humicola was performed in batch and continuous modes in different cultivating system arrangements to compare biomass and carotenoids’ concentration and their productivities. Batch result from stirred tank and airlift photobioreactors indicated the positive effect of increasing light intensity on growth and carotenoid production, whereas the finding from continuous cultivation indicated that carotenoid enhancement preferred high light intensity and nitrogen-deficient environment. The highest biomass (1.31?±?0.04?g?L^?1) and carotenoid (4.59?±?0.06?mg?L^?1) concentration as well as the highest productivities, 0.46?g?L^?1 d^?1 for biomass and 1.61?mg?L^?1 d^?1 for carotenoids, were obtained when maintaining high light intensity of 10 klx, BG-11 medium and 2% (v/v) CO₂ simultaneously, while the highest carotenoid content (4.84?mg?g^?1) was associated with high light intensity and nitrogen-deficient environment, which was induced by feed-modified BG-11 growth medium containing nitrate 20 folds lower than the original medium. Finally, the cultivating system arranged into smaller stirred tank photobioreactors in series yielded approximately 2.5 folds increase in both biomass and carotenoid productivities relative to using single airlift photobioreactor with equivalent working volume and similar operating condition.     

Nutrient removal and biomass production: advances in microalgal biotechnology for wastewater treatment

Owing to certain drawbacks, such as energy-intensive operations in conventional modes of wastewater treatment (WWT), there has been an extensive search for alternative strategies in treatment technology. Biological modes for treating wastewaters are one of the finest technologies in terms of economy and efficiency. An integrated biological approach with chemical flocculation is being conventionally practiced in several-sewage and effluent treatment plants around the world. Overwhelming responsiveness to treat wastewaters especially by using microalgae is due to their simplest photosynthetic mechanism and ease of acclimation to various habitats. Microalgal technology, also known as phycoremediation, has been in use for WWT since 1950s. Various strategies for the cultivation of microalgae in WWT systems are evolving faster. However, the availability of innovative approaches for maximizing the treatment efficiency, coupled with biomass productivity, remains the major bottleneck for commercialization of microalgal technology. Investment costs and invasive parameters also delimit the use of microalgae in WWT. This review critically discusses the merits and demerits of microalgal cultivation strategies recently developed for maximum pollutant removal as well as biomass productivity. Also, the potential of algal biofilm technology in pollutant removal, and harvesting the microalgal biomass using different techniques have been highlighted. Finally, an economic assessment of the currently available methods has been made to validate microalgal cultivation in wastewater at the commercial level.     

Outdoor helical tubular photobioreactors for microalgal production: modeling of fluid-dynamics and mass transfer and assessment of biomass productivity

The production of the microalga Phaeodactylum tricornutum in an outdoor helical reactor was analyzed. First, fluid dynamics, mass-transfer capability, and mixing of the reactor was evaluated at different superficial gas velocities. Performance of the reactor was controlled by power input per culture volume. A maximum liquid velocity of 0.32 m s(-1) and mass transfer coefficient of 0.006 s(-1) were measured at 3200 W m(-3). A model of the influence of superficial gas velocity on the following reactor parameters was proposed: gas hold-up, induced liquid velocity, and mass transfer coefficient, with the accuracy of the model being demonstrated. Second, the influence of superficial gas velocity on the yield of the culture was evaluated in discontinuous and continuous cultures. Mean daily values of culture parameters, including dissolved oxygen, biomass concentration, chlorophyll fluorescence (F(v)/F(m) ratio), growth rate, biomass productivity, and photosynthetic efficiency, were determined. Different growth curves were measured when the superficial gas velocity was modified-the higher the superficial gas velocity, the higher the yield of the system. In continuous mode, biomass productivity increased by 35%, from 1.02 to 1.38 g L(-1) d(-1), when the superficial gas velocity increased from 0.27 to 0.41 m s(-1). Maximal growth rates of 0.068 h(-1), biomass productivities up to 1.4 g L(-1) d(-1), and photosynthetic efficiency of up to 15% were obtained at the higher superficial gas velocity of 0.41 m s(-1). The fluorescence parameter, F(v)/F(m), which reflects the maximal efficiency of PSII photochemistry, showed that the cultures were stressed at average irradiances within the culture higher than 280 microE m(-2) s(-1) at every superficial gas velocity. For nonstressed cultures, the yield of the system was a function of average irradiance inside the culture, with the superficial gas velocity determining this relationship. When superficial gas velocity was increased, higher growth rates, biomass productivities, and photosynthetic efficiencies were obtained for similar average irradiance values. The higher the superficial gas velocity, the higher the liquid velocity, with this increase enhancing the movement of the cells inside the culture. In this way the efficiency of the cells increased and higher biomass concentrations and productivities were reached for the same solar irradiance.     

A novel polyculture growth model of native microalgal communities to estimate biomass productivity for biofuel production

Native polyculture microalgae is a promising scheme to produce microalgal biomass as biofuel feedstock in an open raceway pond. However, predicting biomass productivity of native polycultures microalgae is incredibly complicated. Therefore, developing polyculture growth model to forecast biomass yield is indispensable for commercial-scale production. This research aims to develop a polyculture growth model for native microalgal communities in the Minamisoma algae plant and to estimate biomass and biocrude oil productivity in a semicontinuous open raceway pond. The model was built based on monoculture growth of polyculture species and it is later formulated using species growth, polyculture factor (k_value), initial concentration, light intensity, and temperature. In order to calculate species growth, a simplified Monod model was applied. In the simulation, 115 samples of the 2014–2015 field dataset were used for model training, and 70 samples of the 2017 field dataset were used for model validation. The model simulation on biomass concentration showed that the polyculture growth model with k_value had a root-mean-square error of 0.12, whereas model validation provided a better result with a root-mean-square error of 0.08. Biomass productivity forecast showed maximum productivity of 18.87 g/m²/d in June with an annual average of 13.59 g/m²/d. Biocrude oil yield forecast indicated that hydrothermal liquefaction process was more suitable with a maximum productivity of 0.59 g/m²/d compared with solvent extraction which was only 0.19 g/m²/d. With satisfactory root-mean-square errors less than 0.3, this polyculture growth model can be applied to forecast the productivity of native microalgae.     

Biofuel production from microalgae as feedstock: current status and potential

Algal biofuel has become an attractive alternative of petroleum-based fuels in the past decade. Microalgae have been proposed as a feedstock to produce biodiesel, since they are capable of mitigating CO₂ emission and accumulating lipids with high productivity. This article is an overview of the updated status of biofuels, especially biodiesel production from microalgae including fundamental research, culture selection and engineering process development; it summarizes research on mathematical and life cycle modeling on algae growth and biomass production; and it updates global efforts of research and development and commercialization attempts. The major challenges are also discussed.     

Microalgae for phosphorus removal and biomass production: a six species screen for dual‐purpose organisms

Microalgae biofuel production can be feasible when a second function is added, such as wastewater treatment. Microalgae differ in uptake of phosphorus (P) and growth, making top performer identification fundamental. The objective of this screen was to identify dual‐purpose candidates capable of high rates of P removal and growth. Three freshwater – Chlorella sp., Monoraphidium minutum sp., and Scenedesmus sp. – and three marine – Nannochloropsis sp., N. limnetica sp., and Tetraselmis suecica sp. – species were batch cultured in 250 mL flasks over 16 days to quantitate total phosphorus (TP) removal and growth as a function of P loads (control, and 5, 10, and 15 mg L^?1 enrichment of control). Experimental design used 100 μmol m^?2 s^?1 of light, a light/dark cycle of 14/10 h, and no CO₂ enrichment. Phosphorus uptake was dependent on species, duration of exposure, and treatment, with significant interaction effects. Growth was dependant on species and treatment. Not all species showed increased P removal with increasing P addition, and no species demonstrated higher growth. Nannochloropsis sp and N. limnetica sp. performed poorly across all treatments. Two dual‐purpose candidates were identified. At the 10 mg L^?1 treatment Monoraphidium minutum sp. removed 67.1% (6.66 mg L^?1 ± 0.60 SE) of TP at day 8, 79.3% (7.86 mg L^?1 ± 0.28 SE) at day 16, and biomass accumulation of 0.63 g L^?1 ± 0.06 SE at day 16. At the same treatment Tetraselmis suecica sp. removed 79.4% (6.98 mg L^?1 ± 0.24 SE) TP at day 8, 83.0% (7.30 mg L^?1 ± 0.60 SE) at day 16, and biomass of 0.55 g L^?1 ± 0.02 SE at day 16. These species merit further study using high‐density wastewater cultures and lipid profiling to assess suitability for a nutrient removal and biomass/biofuel production scheme.     

Bioprospecting for oil producing microalgal strains: evaluation of oil and biomass production for ten microalgal strains

Microalgae have the ability to grow rapidly, synthesize and accumulate large amounts (approximately 20-50% of dry weight) of lipids. A successful and economically viable algae based oil industry depends on the selection of appropriate algal strains. In this study ten species of microalgae were prospected to determine their suitability for oil production: Chaetoceros gracilis, Chaetoceros mulleri, Chlorella vulgaris, Dunaliella sp., Isochrysis sp., Nannochloropsis oculata, Tetraselmis sp., Tetraselmis chui, Tetraselmis tetrathele and Thalassiosira weissflogii. The study was carried out in 3 L glass flasks subjected to constant aeration and controlled artificial illumination and temperature at two different salinities. After harvesting, the extraction of oil was carried out using the Bligh and Dyer method assisted by ultrasound. Results showed that C. gracilis presented the highest oil content and that C. vulgaris presented the highest oil production.     

微藻能源技术开发和产业化的发展思路与策略

随着石油资源的日益减少及实现低碳经济的迫切需要,微藻能源已成为世界各国重点研究与发展的战略方向。微藻能源关系国家能源重大战略储备,因此我国迫切需要自主开发微藻能源产业化技术。文中分析了我国发展微藻能源的优势,及目前微藻能源产业化中存在的瓶颈和亟待解决的问题,既包括基础科学研究内容,也包括产业化进程中亟需攻克的关键技术问题。在此基础上,提出微藻能源的发展思路和策略,指出了其产业化中的主要环节的技术发展方向,展望了产业化进程。     

Recent advances in microalgal bioscience in Japan,with special reference to utilization of biomass and metabolites: a review

Japan is one of leading countries in the utilization of and research on microalgae, and various findings have been obtained. Many papers, however, have been published in Japanese, which prevents the information spreading far and wide. The purpose of this review is to introduce recent advances in the utilization of microalgae as well as their basic research in Japan. The discussion covers practical applications ofChlorella andSpirulina biomass to health foods, food additives and feed supplements. The current use of microalgae as live feeds for larvae in aquaculture is also summarized. With respect to microalgal metabolites the present status of research is described with a greater emphasis on bioactive compounds, pigments and oils as potential drugs, coloring matters and biofuels, respectively.     

Land cover changes with the development of anaerobic digestion for biogas production in France

Anaerobic digestion is developing in various countries worldwide to produce renewable energy. In addition, the resulting digestates provide readily available nutrients when applied to cropping systems as fertilizers. The introduction of a biogas plant on a farm can induce land cover changes, in relation to the production of feedstock for the biogas plant and/or to the modification of the farming system. The aim of this study was therefore to characterize and quantify the land cover changes in farms associated with biogas plants in France. We combined two national spatialized databases: the Land Parcel Identification System (yearly French land cover at the parcel scale with farm identifier per parcel) and the SINOE database (biogas plant location and year of start-up). We showed that, on average, the changes were limited, with an increase in maize areas (+3.4% of the total farm areas) compensated by a decrease in wheat and rapeseed areas (−1.8% and −1.9%, respectively), but with a certain variability. The French regulation and market limiting the use of dedicated energy crops seems to have limited land cover changes in France compared to other countries. However, we elaborated a typology of land cover changes and characterized five clusters of farms across the country. The main one (67% of the farms) corresponded to unchanged land cover after the introduction of a biogas plant. The four other clusters showed contrasting changes, for example, an increase or a decrease in grassland areas, a strong increase in maize areas, or a replacement of winter wheat by winter barley. The diversity and the driving factors behind these changes deserve to be better studied and understood through further farmer surveys.     

Evaluation of halophilic microalgae isolated from Rabigh Red Sea coastal area for biodiesel production: Screening and biochemical studies

In the present study, different water samples from Red Sea coastal area at Rabigh city, Saudi Arabia were studied for their dominant algal species. Microalgal isolation was carried out based on dilution method and morphologically examined using F/2 as a growth medium. Dry weight and main biochemical composition (protein, carbohydrates, lipids) of all species were performed at the end of the growth, and biodiesel characteristics were estimated. Nannochloropsis sp., Dunaliella sp., Tetraselmis sp., Prorocentrum sp., Chlorella sp., Nitzschia sp., Coscinodiscus sp., and Navicula sp. were the most dominant species in the collected water samples and were used for further evaluation. Nannochloropsis sp. surpassed all other isolates in concern of biomass production with the maximum recorded dry weight of 0.89 g L^?1, followed by Dunaliella sp. (0.69 g L^?1). The highest crude protein content was observed in Nitzschia sp. (38.21%) and Dunaliella sp. (18.01%), while Nannochloropsis sp. showed 13.38%, with the lowest recorded lipid content in Coscinodiscus sp. (10.09%). Based on the growth, lipid content, and biodiesel characteristics, the present study suggested Dunaliella sp. and Nitzschia sp. as promising candidates for further large-scale biodiesel production.     

A quantitative analysis of microalgal lipids for optimization of biodiesel and omega‐3 production

The lipid characteristics of microalgae are known to differ between species and change with growth conditions. This work provides a methodology for lipid characterization that enables selection of the optimal strain, cultivation conditions, and processing pathway for commercial biodiesel production from microalgae. Two different microalgal species, Nannochloropsis sp. and Chlorella sp., were cultivated under both nitrogen replete and nitrogen depleted conditions. Lipids were extracted and fractionated into three major classes and quantified gravimetrically. The fatty acid profile of each fraction was analyzed using GC–MS. The resulting quantitative lipid data for each of the cultures is discussed in the context of biodiesel and omega‐3 production. This approach illustrates how the growth conditions greatly affect the distribution of fatty acid present in the major lipid classes and therefore the suitability of the lipid extracts for biodiesel and other secondary products. Biotechnol. Bioeng. 2013; 110: 2096–2104. © 2013 Wiley Periodicals, Inc.     

Photosynthetic performance of a cyanobacterium in a vertical flat-plate photobioreactor for outdoor microalgal production and fixation of CO2

A vertical flat-plate photobioreactor was developed for the outdoor culture of microalgae using sunlight as the light source. The ability for biomass production and CO₂ fixation was evaluated by using a cyanobacterium, Synechocystis aquatilis SI-2. The average areal productivity was 31 g biomass m^–2 d^–1, which corresponded to a CO₂ fixation rate of 51 g CO₂ m^–2 d^–1, sustainable in the northern region of Japan during the winter time (January and February). The relationships between the efficiency of solar energy utilization of the reactor and its effect factors (cell concentration and irradiation) were investigated.     

Biotechnology of algal biomass production: a review of systems for outdoor mass culture

Microalgae are very efficient solar energy converters and they can produce a great variety of metabolites. Man has always tried to take advantage of these proporties through algal mass culture. Despite the fact that many applications for microalgae have been described in the literature, these micro-organisms are still of minor economic importance. Industrial reactors for algal culture are at present, all designed as open race-ways (shallow open ponds where culture is circulated by a paddle-wheel). Technical and biological limitations of these open systems have given rise to the development of enclosed photoreactors (made of transparent tubes, sleeves or containers and where light source may be natural or artificial). The present review surveys advances in these two technologies for cultivation of microalgae. Starting from published results, the advantages and disadvantages of open systems and closed photobioreactors are discussed. A few open systems are presented for which particularly reliable results are available. Emphasis is then put on closed systems, which have been considered as capital intensive and are justified only when a fine chemical is to be produced.     

Phycoremediation coupled production of algal biomass,harvesting and anaerobic digestion: Possibilities and challenges

Biogas produced from anaerobic digestion is a versatile and environment friendly fuel which traditionally utilizes cattle dung as the substrate. In the recent years, owing to its high content of biodegradable compounds, algal biomass has emerged as a potential feedstock for biogas production. Moreover, the ability of algae to treat wastewater and fix CO₂ from waste gas streams makes it an environmental friendly and economically feasible feedstock. The present review focuses on the possibility of utilizing wastewater as the nutrient and waste gases as the CO₂ source for algal biomass production and subsequent biogas generation. Studies describing the various harvesting methods of algal biomass as well as its anaerobic digestion have been compiled and discussed. Studies targeting the most recent advancements on biogas enrichment by algae have been discussed. Apart from highlighting the various advantages of utilizing algal biomass for biogas production, limitations of the process such as cell wall resistivity towards digestion and inhibitions caused due to ammonia toxicity and the possible strategies for overcoming the same have been reviewed. The studies compiled in the present review indicate that if the challenges posed in translating the lab scale studies on phycoremediation and biogas production to pilot scale are overcome, algal biogas could become the sustainable and economically feasible source of renewable energy.     

Kimchi microflora: history,current status,and perspectives for industrial kimchi production

Kimchi, a traditional Korean food made by the fermentation of vegetables, has become popular globally because of its organoleptic, beneficial, and nutritional properties. Spontaneous kimchi fermentation in unsterilized raw materials leads to the growth of various lactic acid bacteria (LAB), which results in variations in the taste and sensory qualities of kimchi products and difficulties in the standardized industrial production of kimchi. Raw materials, kimchi varieties, ingredients, and fermentation conditions have significant effects on the microbial communities and fermentative characteristics of kimchi during fermentation. Heterofermentative LAB belonging to the genera Leuconostoc, Lactobacillus, and Weissella are likely to be key players in kimchi fermentation and have been subjected to genomic and functional studies to gain a better understanding of the fermentation process and beneficial effects of kimchi. The use of starter cultures has been considered for the industrial production of high quality, standardized kimchi. Here, we review the composition and biochemistry of kimchi microflora communities, functional and genomic studies of kimchi LAB, and perspectives for industrial kimchi production.     

Energy and greenhouse gas balance of bioenergy production from poplar and willow: a review

Short‐rotation woody crops (SRWC) such as poplar and willow are an important source of renewable energy. They can be converted into electricity and/or heat using conventional or modern biomass technologies. In recent years many studies have examined the energy and greenhouse gas (GHG) balance of bioenergy production from poplar and willow using various approaches. The outcomes of these studies have, however, generated controversy among scientists, policy makers, and the society. This paper reviews 26 studies on energy and GHG balance of bioenergy production from poplar and willow published between 1990 and 2009. The data published in the reviewed literature gave energy ratios (ER) between 13 and 79 for the cradle‐to‐farm gate and between 3 and 16 for cradle‐to‐plant assessments, whereas the intensity of GHG emissions ranged from 0.6 to 10.6 g CO₂ Eq MJ_biomass^?1 and 39 to 132 g CO₂ Eq kWh^?1. These values vary substantially among the reviewed studies depending on the system boundaries and methodological assumptions. The lack of transparency hampers meaningful comparisons among studies. Although specific numerical results differ, our review revealed a general consensus on two points: SRWC yielded 14.1–85.9 times more energy than coal (ER_coal～0.9) per unit of fossil energy input, and GHG emissions were 9–161 times lower than those of coal (GHG_coal～96.8). To help to reduce the substantial variability in results, this review suggests a standardization of the assumptions about methodological issues. Likewise, the development of a widely accepted framework toward a reliable analysis of energy in bioenergy production systems is most needed.     

Thermophilic,anaerobic co-digestion of microalgal biomass and cellulose for H<Subscript>2</Subscript> production

Microalgal biomass has been a focus in the sustainable energy field, especially biodiesel production. The purpose of this study was to assess the feasibility of treating microalgal biomass and cellulose by anaerobic digestion for H₂ production. A microbial consortium, TC60, known to degrade cellulose and other plant polymers, was enriched on a mixture of cellulose and green microalgal biomass of Dunaliella tertiolecta, a marine species, or Chlorella vulgaris, a freshwater species. After five enrichment steps at 60°C, hydrogen yields increased at least 10% under all conditions. Anaerobic digestion of D. tertiolecta and cellulose by TC60 produced 7.7 mmol H₂/g volatile solids (VS) which were higher than the levels (2.9–4.2 mmol/g VS) obtained with cellulose and C. vulgaris biomass. Both microalgal slurries contained satellite prokaryotes. The C. vulgaris slurry, without TC60 inoculation, generated H₂ levels on par with that of TC60 on cellulose alone. The biomass-fed anaerobic digestion resulted in large shifts in short chain fatty acid concentrations and increased ammonium levels. Growth and H₂ production increased when TC60 was grown on a combination of D. tertiolecta and cellulose due to nutrients released from algal cells via lysis. The results indicated that satellite heterotrophs from C. vulgaris produced H₂ but the Chlorella biomass was not substantially degraded by TC60. To date, this is the first study to examine H₂ production by anaerobic digestion of microalgal biomass. The results indicate that H₂ production is feasible but higher yields could be achieved by optimization of the bioprocess conditions including biomass pretreatment.     

Stirring and biomass starter influences the anaerobic digestion of different substrates for biogas production

Here, we present the results of lab‐scale experiments conducted in a batch mode to determine the biogas yield of lipid‐rich waste and corn silage under the effect of stirring. Further semi‐continuous experiments were carried out for the lipid‐rich waste with/without stirring. Additionally, it was analyzed how the starter used for the batch experiment influences the digestion process. The results showed a significant stirring effect on the anaerobic digestion only when seed sludge from a biogas plant was used as a starter. In this case, the experiments without stirring yielded only about 50% of the expected biogas for the investigated substrates. The addition of manure slurry to the batch reactor as part of the starter improved the biogas production. The more diluted media in the reactor allowed a better contact between the bacteria and the substrates making stirring not necessary.