Bioprospecting for hyper-lipid producing microalgal strains for sustainable biofuel production

Global petroleum reserves are shrinking at a fast pace, increasing the demand for alternate fuels. Microalgae have the ability to grow rapidly, and synthesize and accumulate large amounts (approximately 20-50% of dry weight) of neutral lipid stored in cytosolic lipid bodies. A successful and economically viable algae based biofuel industry mainly depends on the selection of appropriate algal strains. The main focus of bioprospecting for microalgae is to identify unique high lipid producing microalgae from different habitats. Indigenous species of microalgae with high lipid yields are especially valuable in the biofuel industry. Isolation, purification and identification of natural microalgal assemblages using conventional techniques is generally time consuming. However, the recent use of micromanipulation as a rapid isolating tool allows for a higher screening throughput. The appropriate media and growth conditions are also important for successful microalgal proliferation. Environmental parameters recorded at the sampling site are necessary to optimize in vitro growth. Identification of species generally requires a combination of morphological and genetic characterization. The selected microalgal strains are grown in upscale systems such as raceway ponds or photobireactors for biomass and lipid production. This paper reviews the recent methodologies adopted for site selection, sampling, strain selection and identification, optimization of cultural conditions for superior lipid yield for biofuel production. Energy generation routes of microalgal lipids and biomass are discussed in detail.     

Closed photobioreactors for production of microalgal biomasses

Microalgal biomasses have been produced industrially for a long history for application in a variety of different fields. Most recently, microalgae are established as the most promising species for biofuel production and CO(2) bio-sequestration owing to their high photosynthesis efficiency. Nevertheless, design of photobioreactors that maximize solar energy capture and conversion has been one of the major challenges in commercial microalga biomass production. In this review, we systematically survey the recent developments in this field.     

Extraction of oil from microalgae for biodiesel production: A review

The rapid increase of CO(2) concentration in the atmosphere combined with depleted supplies of fossil fuels has led to an increased commercial interest in renewable fuels. Due to their high biomass productivity, rapid lipid accumulation, and ability to survive in saline water, microalgae have been identified as promising feedstocks for industrial-scale production of carbon-neutral biodiesel. This study examines the principles involved in lipid extraction from microalgal cells, a crucial downstream processing step in the production of microalgal biodiesel. We analyze the different technological options currently available for laboratory-scale microalgal lipid extraction, with a primary focus on the prospect of organic solvent and supercritical fluid extraction. The study also provides an assessment of recent breakthroughs in this rapidly developing field and reports on the suitability of microalgal lipid compositions for biodiesel conversion.     

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.     

Micro and macroalgal biomass: A renewable source for bioethanol

Population outburst together with increased motorization has led to an overwhelming increase in the demand for fuel. In the milieu of economical and environmental concern, algae capable of accumulating high starch/cellulose can serve as an excellent alternative to food crops for bioethanol production, a green fuel for sustainable future. Certain species of algae can produce ethanol during dark-anaerobic fermentation and thus serve as a direct source for ethanol production. Of late, oleaginous microalgae generate high starch/cellulose biomass waste after oil extraction, which can be hydrolyzed to generate sugary syrup to be used as substrate for ethanol production. Macroalgae are also harnessed as renewable source of biomass intended for ethanol production. Currently there are very few studies on this issue, and intense research is required in future in this area for efficient utilization of algal biomass and their industrial wastes to produce environmentally friendly fuel bioethanol.     

Two-stage heterotrophic and phototrophic culture strategy for algal biomass and lipid production

A two-stage heterotrophic and phototrophic culture strategy for algal biomass and lipid production was studied, wherein high density heterotrophic cultures of Chlorellasorokiniana serve as seed for subsequent phototrophic growth. The data showed growth rate, cell density and productivity of heterotrophic C.sorokiniana were 3.0, 3.3 and 7.4 times higher than phototrophic counterpart, respectively. Hetero- and phototrophic algal seeds had similar biomass/lipid production and fatty acid profile when inoculated into phototrophic culture system. To expand the application, food waste and wastewater were tested as feedstock for heterotrophic growth, and supported cell growth successfully. These results demonstrated the advantages of using heterotrophic algae cells as seeds for open algae culture system. Additionally, high inoculation rate of heterotrophic algal seed can be utilized as an effective method for contamination control. This two-stage heterotrophic phototrophic process is promising to provide a more efficient way for large scale production of algal biomass and biofuels.     

Oil extraction from microalgae for biodiesel production

This study examines the performance of supercritical carbon dioxide (SCCO₂) extraction and hexane extraction of lipids from marine Chlorococcum sp. for lab-scale biodiesel production. Even though the strain of Chlorococcum sp. used in this study had a low maximum lipid yield (7.1 wt% to dry biomass), the extracted lipid displayed a suitable fatty acid profile for biodiesel [C18:1 (∼63 wt%), C16:0 (∼19 wt%), C18:2 (∼4 wt%), C16:1 (∼4 wt%), and C18:0 (∼3 wt%)]. For SCCO₂ extraction, decreasing temperature and increasing pressure resulted in increased lipid yields. The mass transfer coefficient (k) for lipid extraction under supercritical conditions was found to increase with fluid dielectric constant as well as fluid density. For hexane extraction, continuous operation with a Soxhlet apparatus and inclusion of isopropanol as a co-solvent enhanced lipid yields. Hexane extraction from either dried microalgal powder or wet microalgal paste obtained comparable lipid yields.     

Systematic investigation of biomass and lipid productivity by microalgae in photobioreactors for biodiesel application

We describe a methodology to investigate the potential of given microalgae species for biodiesel production by characterizing their productivity in terms of both biomass and lipids. A multi-step approach was used: determination of biological needs for macronutrients (nitrate, phosphate and sulphate), determination of maximum biomass productivity (the “light-limited” regime), scaling-up of biomass production in photobioreactors, including a theoretical framework to predict corresponding productivities, and investigation of how nitrate starvation protocol affects cell biochemical composition and triggers triacylglycerol (TAG) accumulation. The methodology was applied to two freshwater strains, Chlorella vulgaris and Neochloris oleoabundans, and one seawater diatom strain, Cylindrotheca closterium. The highest total lipid content was achieved with N. oleoabundans (25-37% of DW), while the highest TAG content was found in C. vulgaris (11-14% of DW). These two species showed similar TAG productivities.     

Local bioprospecting for high-lipid producing microalgal strains to be grown on concentrated municipal wastewater for biofuel production

Mass cultivation of microalgae for biofuel production depends heavily on the performance of the microalgae strains used. In this study, 60 algae-like microorganisms collected from different sampling sites in Minnesota were examined using multi-step screening and acclimation procedures to select high-lipid producing facultative heterotrophic microalgae strains capable of growing on concentrated municipal wastewater (CMW) for simultaneous energy crop production and wastewater treatment. Twenty-seven facultative heterotrophic microalgae strains were found, among which 17 strains were proved to be tolerant to CMW. These 17 top-performing strains were identified through morphological observation and DNA sequencing as Chlorella sp., Heynigia sp., Hindakia sp., Micractinium sp., and Scenedesmus sp. Five strains were chosen for other studies because of their ability to adapt to CMW, high growth rates (0.455-0.498 d⁻¹) and higher lipid productivities (74.5-77.8 mg L⁻¹ d⁻¹). These strains are considered highly promising compared with other strains reported in the literature.     

Advances in direct transesterification of microalgal biomass for biodiesel production

Microalgae biomass is becoming an interesting raw material to produce biodiesel, where several approaches in the transesterification process have been applied such as different catalysts, different acyl acceptors, incorporation of co-solvents and the different operational conditions. However there are some drawbacks that must be solved before any industrial application could be intended. The main problems are related with the high water content of the biomass (over 80 %) and the several process steps involved in biodiesel production such as: drying, cell disruption, oils extraction, transesterification and biodiesel refining. In comparison to other alternatives, the use of direct transesterification could be a suitable alternative since cell disruption, lipids extraction and transesterification are carried out in one step, with a direct reaction of oil-bearing biomass to biodiesel. This process could be applied even using biomass with high water content, and its efficiency could be improved by the incorporation of promising technologies such as microwave or ultrasonication that can enhance the mass transfer rate between immiscible phases, simultaneously diminishing the reaction time. However, it is still necessary to decrease the costs of these technologies so they can be suitable alternatives in future industrial applications.     

Aqueous ammonia pretreatment of oil palm empty fruit bunches for ethanol production

Oil palm empty fruit bunches (EFB) were pretreated by aqueous ammonia soaking for ethanol production. Pretreated EFB, which were pretreated at the optimal conditions of 60 °C, 12 h, and 21% (w/w) aqueous ammonia, showed 19.5% and 41.4% glucose yields during an enzymatic digestibility test for 96 h when using 15 and 60 FPU of cellulase, respectively. Using the pretreated EFB, simultaneous saccharification and fermentation for 168 h with 5% (w/v) glucan loading and 60 FPU of cellulase and 30 CBU of β-glucosidase per gram glucan resulted in ethanol production of 18.6 g/L titer, 65.6% of theoretical maximum yield, and 0.11 g/L/h of productivity.     

Properties of oil and char derived from slow pyrolysis of Tetraselmis chui

Pyrolysis of biomass is a means to industrially manufacture renewable oil and gas, in addition to biochar for soil amendment and long-term carbon fixation. In this work, oil and char derived from the slow pyrolysis of the unicellular marine diatom Tetraselmis chui are analysed using a variety of techniques. The pyrolytic oil fraction exhibits a wide variety of fatty acids, alkanes, alkenes, amides, aldehydes, terpenes, pyrrolidinines, phytol and phenols, with a high heating value (HHV) of 28 MJ/kg. The biochar produced has a HHV of 14.5 MJ/kg and reveals a number of properties that are potentially valuable from an agronomic point of view, including high cation exchange capacity (CEC), large concentration of N, and a low C:N ratio. The quantity of C in T. chui biochar that can be expected to stabilise in soil amounts to approximately 9%/wt of the original feedstock, leading to a potential net reduction in atmospheric CO₂.     

Microalgae biodiversity and biomass status in Qua Iboe Estuary mangrove swamp,Nigeria

Microalgae composition, abundance, diversity and biomass of the Qua Iboe Estuary mangrove swamp were studied. The results revealed the rich assemblage of the brackish ecosystem. Six major taxanomic classes were encountered. These were the Bacillariophyceae, Cyanophyceae, Chlorophyceae, Chrysophyceae, Euglenophyceae and Phaeophyceae. Their composition, abundance and diversity exhibits strong seasonal variation. Variations between pelagic and sedimentary habitats were also noticed. The diatoms (Bacillariophyceae) dominated the habitats. Actinoptychus undalatus, Navicula radiosa and Amphora ovalis co-dominated the pelagic water column; while the epipellic (intertidal) and benthic (subtidal) sediments were co-dominated by A. ovalis and Actinoptychus undulatus. Analyses of the Shannon’s index of general diversity (H¹), McArthur-Terborgh species equitability index (E) and Simpson’s index of dominance (D) of the microalgae communities revealed that A. ovalis with a mean H ¹ -value of 0.25, E-value of 0.08 and D-value of 0.05 was the most prevalent genus in the mangrove ecosystem despite its uneven distribution. The densities of the microalgae communities corresponded with their biomass statuses and were seriously impacted by oil spillage. This forms the basis of concern because the estuary is associated with a high probability of major oil pollution with serious consequences for ecological stability and fisheries.     

Cultivation, photobioreactor design and harvesting of microalgae for biodiesel production: A critical review

Microalgae have the ability to mitigate CO₂ emission and produce oil with a high productivity, thereby having the potential for applications in producing the third-generation of biofuels. The key technologies for producing microalgal biofuels include identification of preferable culture conditions for high oil productivity, development of effective and economical microalgae cultivation systems, as well as separation and harvesting of microalgal biomass and oil. This review presents recent advances in microalgal cultivation, photobioreactor design, and harvesting technologies with a focus on microalgal oil (mainly triglycerides) production. The effects of different microalgal metabolisms (i.e., phototrophic, heterotrophic, mixotrophic, and photoheterotrophic growth), cultivation systems (emphasizing the effect of light sources), and biomass harvesting methods (chemical/physical methods) on microalgal biomass and oil production are compared and critically discussed. This review aims to provide useful information to help future development of efficient and commercially viable technology for microalgae-based biodiesel production.     

Integration of microalgal cultivation system for wastewater remediation and sustainable biomass production

Untreated wastewaters have been a great concern and can cause major pollution problems for environment. Conventional approaches for treating wastewater involve tremendous capital cost, have major short comings and are not sustainable. Microalgae culture offers an interesting step for wastewater treatment. Microalgae serve the dual purpose of phycoremediation along with the production of potentially valuable biomass, which can be used for several purposes. The ability of microalgae to accumulate nitrogen, phosphorus, heavy metals and other toxic compounds can be integrated with wastewater treatment system to offer an elegant solution towards tertiary and quaternary treatment. The current review explores possible role of microalgal based wastewater treatment and explores the current progress, key challenges, limitations and future prospects with special emphasis on strategies involved in harvesting, boosting biomass and lipid yield.     

Optimization of the biomass production of oil algae Chlorella minutissima UTEX2341

High production cost is a major obstacle to the extensive use of microalgae biodiesel. To cut the cost and achieve higher biomass productivity, Chlorella minutissima UTEX2341 was cultured under photoheterotrophic conditions. With the carbon, nitrogen and phosphorus concentration of 26.37, 2.61 and 0.03 g L?1 d?1 respectively, a maximum biomass productivity of 1.78 g L?1 d?1 was obtained, which was 59 times more than that cultured under autotrophic condition. The lipid productivity reached 0.29 g L?1 d?1, which was 11.9 times higher than the highest value reported by Oh et al. (2010). The conversion rate of microalgae lipids to FAME was found to be elevated from 45.65% to 62.97% and the FAME productivity increased from 1.16 to 180.68 mg L?1 d?1 after the optimization. 94% of the fatty acid of C. minutissima UTEX2341 was found to be composed of palmitic, oleic, linoleic and γ linoleic and the unsaturated fatty acids were the main parts (79.42%).     

Two-phase systems: potential for in situ extraction of microalgal products

Algae are currently used for production of niche products and are becoming increasingly interesting for the production of bulk commodities, such as biodiesel. For the production of these goods to become economically feasible, production costs will have to be lowered by one order of magnitude. The application of two-phase systems could be used to lower production costs. These systems circumvent the costly step of cell harvesting, whilst the product is extracted and prepared for downstream processing. The mechanism of extraction is a fundamental aspect of the practical question whether two-phase systems can be applied for in situ extraction, viz, simultaneous growth, product formation and extraction, or as a separate downstream processing step. Three possible mechanisms are discussed; 1) product excretion 2) cell permeabilization, and 3) cell death. It was shown that in the case of product excretion, the application of two-phase systems for in situ extraction can be very valuable. With permeabilization and cell death, in situ extraction is not ideal, but the application of two-phase systems as downstream extraction steps can be part of a well-designed biorefinery process. In this way, processing costs can be decreased while the product is mildly and selectively extracted.Thus far none of the algal strains used in two-phase systems have been shown to excrete their product; the output has always been the result of cell death. Two-phase systems can be a good approach as a downstream processing step for these species. For future applications of two-phase in situ extraction in algal production processes, either new species that show product excretion should be discovered, or existing species should be modified to induce product excretion.     

The potential of microalgal biomass production for biotechnological purposes using wastewater resources

The utilization of microalgae for wastewater treatment represents an attractive opportunity for wastewater valorization through the use of the produced biomass. Five strains of microalgae were isolated from municipal wastewater and grown in autoclaved and non-autoclaved effluent at 30 °C and 150 μmol photons m^?2 s^?1 to study biomass production, nutrient removal, and the biochemical composition of the biomass. All strains reached high biomass productivity (35.6 to 54.2 mg dry weight L^?1 day^?1) within 4 days of batch culturing. In this period, ammonium-N and phosphate were reduced by more than 60 and 90 %, respectively. The high growth rate (0.57 to 1.06 day^?1) ensured a rapid removal of nutrients and thereby a short retention time. By the fourth day of cultivation, the algal biomass contained 32 % protein, but only 11 % lipids and 18 % carbohydrates. It was found that the biomass was a suitable raw material for biogas production by anaerobic digestion. Biodigestion of obtained biomass was simulated by employing the Aspen HYSYS modeling software, resulting in methane yields comparable to those found in the literature. The elemental analysis of the algal biomass showed very low concentrations of pollutants, demonstrating the potential of use of the digestate from biodigestion as a bio-fertilizer.     

Simultaneous flue gas bioremediation and reduction of microalgal biomass production costs

A flue gas originating from a municipal waste incinerator was used as a source of CO₂ for the cultivation of the microalga Chlorella vulgaris, in order to decrease the biomass production costs and to bioremediate CO₂ simultaneously. The utilization of the flue gas containing 10–13% (v/v) CO₂ and 8–10% (v/v) O₂ for the photobioreactor agitation and CO₂ supply was proven to be convenient. The growth rate of algal cultures on the flue gas was even higher when compared with the control culture supplied by a mixture of pure CO₂ and air (11% (v/v) CO₂). Correspondingly, the CO₂ fixation rate was also higher when using the flue gas (4.4 g CO₂ l⁻¹ 24 h⁻¹) than using the control gas (3.0 g CO₂ l⁻¹ 24 h⁻¹). The toxicological analysis of the biomass produced using untreated flue gas showed only a slight excess of mercury while all the other compounds (other heavy metals, polycyclic aromatic hydrocarbons, polychlorinated dibenzodioxins and dibenzofurans, and polychlorinated biphenyls) were below the limits required by the European Union foodstuff legislation. Fortunately, extending the flue gas treatment prior to the cultivation unit by a simple granulated activated carbon column led to an efficient absorption of gaseous mercury and to the algal biomass composition compliant with all the foodstuff legislation requirements.     

The potential of sustainable algal biofuel production using wastewater resources

The potential of microalgae as a source of renewable energy has received considerable interest, but if microalgal biofuel production is to be economically viable and sustainable, further optimization of mass culture conditions are needed. Wastewaters derived from municipal, agricultural and industrial activities potentially provide cost-effective and sustainable means of algal growth for biofuels. In addition, there is also potential for combining wastewater treatment by algae, such as nutrient removal, with biofuel production. Here we will review the current research on this topic and discuss the potential benefits and limitations of using wastewaters as resources for cost-effective microalgal biofuel production.