Microalgal carbohydrates: an overview of the factors influencing carbohydrates production, and of main bioconversion technologies for production of biofuels

Microalgal biomass seems to be a promising feedstock for biofuel generation. Microalgae have relative high photosynthetic efficiencies, high growth rates, and some species can thrive in brackish water or seawater and wastewater from the food- and agro-industrial sector. Today, the main interest in research is the cultivation of microalgae for lipids production to generate biodiesel. However, there are several other biological or thermochemical conversion technologies, in which microalgal biomass could be used as substrate. However, the high protein content or the low carbohydrate content of the majority of the microalgal species might be a constraint for their possible use in these technologies. Moreover, in the majority of biomass conversion technologies, carbohydrates are the main substrate for production of biofuels. Nevertheless, microalgae biomass composition could be manipulated by several cultivation techniques, such as nutrient starvation or other stressed environmental conditions, which cause the microalgae to accumulate carbohydrates. This paper attempts to give a general overview of techniques that can be used for increasing the microalgal biomass carbohydrate content. In addition, biomass conversion technologies, related to the conversion of carbohydrates into biofuels are discussed.     

The development of microalgal biotechnology in the Czech Republic

Microscopic algae and cyanobacteria are excellent sources of numerous compounds, from raw biomass rich in proteins, oils, and antioxidants to valuable secondary metabolites with potential medical use. In the former Czechoslovakia, microalgal biotechnology developed rapidly in the 1960s with the main aim of providing industrial, high-yield sources of algal biomass. Unique cultivation techniques that are still in use were successfully developed and tested. Gradually, the focus changed from bulk production to more sophisticated use of microalgae, including production of bioactive compounds. Along the way, better understanding of the physiology and cell biology of productive microalgal strains was achieved. Currently, microalgae are in the focus again, mostly as possible sources of bioactive compounds and next-generation biofuels for the 21st century.     

Microalgae-based carbohydrates for biofuel production

Microalgae are considered as the most promising renewable feedstock for biofuel production and biorefineries, due to their advantages of fast growth, efficient carbon dioxide fixation, not competing for arable lands and potable water, and potentially accumulating high amounts of lipids and carbohydrates. Since carbohydrates in microalgae biomass are mainly cellulose in the cell wall and starch in the plastids without lignin and low hemicelluloses contents, they can be readily converted into fermentable sugars. However, to date there are very few studies focusing on the use of microalgae-based carbohydrates for biofuel production, which requires more understanding and knowledge to support the technical feasibility of this next-generation feedstock. This review article elucidates comprehensive information on the characteristics and metabolism of main fermentable microalgal carbohydrates (e.g., starch and cellulose), as well as the key factors and challenges that should be addressed during production and saccharification of microalgal carbohydrates. Furthermore, developments on the utilization of microalgae-based feedstock in producing liquid and gaseous biofuels are summarized. The objective of this article is to provide useful knowledge and information with regard to biochemistry, bioprocess engineering, and commercial applications to assist in the viable technology development of for biofuels generation from microalgae-based carbohydrates.     

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.     

Microalgae potential as a biogas source: current status,restraints and future trends

In recent years, the world energy demands have had a recurrent increase. For this reason the alternative to the fossil fuel resources are trend topics in investigation. Microalgae have been extensively studied as a source of biofuels and as one of the most promising alternatives in this new framework. One of the possibilities of obtaining renewable energy from microalgae is biogas production using anaerobic digestion process. This process is considered a significant component for biofuels and waste management, since represent an opportunity for energy generation using different wastewater products; also, the economic viability of microalgae liquid biofuel production could be improved. However, the anaerobic digestion of microalgae biomass is still not optimized because of the numerous technical limitations such as the microalgae characteristics, low carbon:nitrogen ratio, ammonia toxicity and even salinity. The present review summarizes and compares information concerning to anaerobic digestion of microalgal biomass and future directions for research. Besides, specific operational factors and potential inhibitory parameters of the process are analyzed and compared. Additionally, the paper covers the state or art concerning in methane production enhancement from algal biomass.     

Current advances on fermentative biobutanol production using third generation feedstock

Biobutanol is gaining more attention as a potential alternative to ethanol, and the demand for fermentative biobutanol production has renewed interest. The main challenge faced in biobutanol production is the availability of feedstock. Using conventional agricultural biomass as feedstock is controversial and less efficient, while microalgae, the third generation feedstock, are considered promising feedstock for biobutanol production due to their high growth rate and high carbohydrates content. This review is primarily focused on biobutanol production by using carbohydrate-rich microalgal feedstock. Key technologies and challenges involved in producing butanol from microalgae are discussed in detail and future directions are also presented.     

Anaerobic digestion of microalgal biomass for bioenergy production,removal of nutrients and microcystin: current status

Using renewable microalgal biomass as active feedstocks for biofuels and bioproducts is explored to substitute petroleum-based fuels and chemicals. In the last few years, the importance of microalgae biomass has been realized as a renewable feedstock due to several positive attributes associated with it. Biorefinery via anaerobic digestion (AD) of microalgal biomass is a promising and sustainable method to produce value-added chemicals, edible products and biofuels. Microalgal biomass pretreatment is a significant process to enhance methane production by AD. Findings on the AD microbial community’s variety and organization can give novel in turn on digester steadiness and presentation. This review presents a vital study of the existing facts on the AD microbial community and AD production. Co-digestion of microalgal biomass with different co-substrates was used in AD to enhance biogas production, and the process was economically viable with improved biodegradability. Microcystins, which are produced by toxic cyanobacterial blooms, create a severe hazard to environmental health. Anaerobic biodegradation is an effective method to degrade the microcystins and convert into nontoxic products. However, for the cost-effective conversion of biomass to energy and other beneficial byproducts, additional highly developed research is still required for large-scale AD of microalgal biomass.     

Cyanobacteria and microalgae: a positive prospect for biofuels

Biofuel-bioenergy production has generated intensive interest due to increased concern regarding limited petroleum-based fuel supplies and their contribution to atmospheric CO2 levels. Biofuel research is not just a matter of finding the right type of biomass and converting it to fuel, but it must also be economically sustainable on large-scale. Several aspects of cyanobacteria and microalgae such as oxygenic photosynthesis, high per-acre productivity, non-food based feedstock, growth on non-productive and non-arable land, utilization of wide variety of water sources (fresh, brackish, seawater and wastewater) and production of valuable co-products along with biofuels have combined to capture the interest of researchers and entrepreneurs. Currently, worldwide biofuels mainly in focus include biohydrogen, bioethanol, biodiesel and biogas. This review focuses on cultivation and harvesting of cyanobacteria and microalgae, possible biofuels and co-products, challenges for cyanobacterial and microalgal biofuels and the approaches of genetic engineering and modifications to increase biofuel production.     

Methods of downstream processing for the production of biodiesel from microalgae

Despite receiving increasing attention during the last few decades, the production of microalgal biofuels is not yet sufficiently cost-effective to compete with that of petroleum-based conventional fuels. Among the steps required for the production of microalgal biofuels, the harvest of the microalgal biomass and the extraction of lipids from microalgae are two of the most expensive. In this review article, we surveyed a substantial amount of previous work in microalgal harvesting and lipid extraction to highlight recent progress in these areas. We also discuss new developments in the biodiesel conversion technology due to the importance of the connectivity of this step with the lipid extraction process. Furthermore, we propose possible future directions for technological or process improvements that will directly affect the final production costs of microalgal biomass-based biofuels.     

Microalgae for high-value compounds and biofuels production: A review with focus on cultivation under stress conditions

Microalgal biomass as feedstock for biofuel production is an attracting alternative to terrestrial plant utilization for biofuels production. However, today the microalgal cultivation systems for energy production purposes seem not yet to be economically feasible. Microalgae, though cultivated under stress conditions, such as nutrient starvation, high salinity, high temperature etc. accumulate considerable amounts (up to 60–65% of dry weight) of lipids or carbohydrates along with several secondary metabolites. Especially some of the latter are valuable compounds with an enormous range of industrial applications. The simultaneous production of lipids or carbohydrates for biofuel production and of secondary metabolites in a biorefinery concept might allow the microalgal production to be economically feasible. This paper aims to provide a review on the available literature about the cultivation of microalgae for the accumulation of high-value compounds along with lipids or carbohydrates focusing on stress cultivation conditions.     

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.     

微藻异养高密度培养研究进展与发展趋势*

微藻细胞可以积累大量油脂、蛋白质、多糖、色素、不饱和脂肪酸等物质,在能源、食品、饵料、保健品及药品等行业有巨大的应用价值。然而,微藻在传统光自养模式下很难实现高密度培养来大量生产这些重要的物质,进而限制了微藻的实际应用。相反,微藻在异养模式下生长速度快、生物质浓度高,可以短时间内获得大量微藻生物质。因此,异养高密度培养微藻具备大规模、高效率培养微藻生产目标产物的巨大潜力。阐述微藻异养培养的优缺点及相应技术难点的解决思路、影响微藻异养生长及目标产物积累的主要营养因子和环境因子、微藻异养高密度培养的方式及微藻异养高密度培养的当前发展水平。结合文献报道分析微藻异养高密度培养的四个具有极大发展潜力的发展方向,以期更好地利用异养模式来高效率、低成本培养微藻生产大量目标产物,满足上述多个行业对微藻原材料的巨大需求,从而加速微藻产业的发展。     

The dynamics of heterotrophic algal cultures

In this work, the time varying characteristics of microalgal cultures are investigated. Microalgae are a promising source of biofuels and other valuable chemicals; a better understanding of their dynamic behavior is, however, required to facilitate process scale-up, optimization and control. Growth and oil production rates are evaluated as a function of carbon and nitrogen sources concentration. It is found that nitrogen has a major role in controlling the productivity of microalgae. Moreover, it is shown that there exists a nitrogen source concentration at which biomass and oil production can be maximized. A mathematical model that describes the effect of nitrogen and carbon source on growth and oil production is proposed. The model considers the uncoupling between nutrient uptake and growth, a characteristic of algal cells. Validity of the proposed model is tested on fed-batch cultures.     

Biodiesel production with microalgae as feedstock: from strains to biodiesel

Due to negative environmental influence and limited availability, petroleum-derived fuels need to be replaced by renewable biofuels. Biodiesel has attracted intensive attention as an important biofuel. Microalgae have numerous advantages for biodiesel production over many terrestrial plants. There are a series of consecutive processes for biodiesel production with microalgae as feedstock, including selection of adequate microalgal strains, mass culture, cell harvesting, oil extraction and transesterification. To reduce the overall production cost, technology development and process optimization are necessary. Genetic engineering also plays an important role in manipulating lipid biosynthesis in microalgae. Many approaches, such as sequestering carbon dioxide from industrial plants for the carbon source, using wastewater for the nutrient supply, and maximizing the values of by-products, have shown a potential for cost reduction. This review provides a brief overview of the process of biodiesel production with microalgae as feedstock. The methods associated with this process (e.g. lipid determination, mass culture, oil extraction) are also compared and discussed.     

Integration of microalgae cultivation with industrial waste remediation for biofuel and bioenergy production: opportunities and limitations

There is currently a renewed interest in developing microalgae as a source of renewable energy and fuel. Microalgae hold great potential as a source of biomass for the production of energy and fungible liquid transportation fuels. However, the technologies required for large-scale cultivation, processing, and conversion of microalgal biomass to energy products are underdeveloped. Microalgae offer several advantages over traditional 'first-generation' biofuels crops like corn: these include superior biomass productivity, the ability to grow on poor-quality land unsuitable for agriculture, and the potential for sustainable growth by extracting macro- and micronutrients from wastewater and industrial flue-stack emissions. Integrating microalgal cultivation with municipal wastewater treatment and industrial CO(2) emissions from coal-fired power plants is a potential strategy to produce large quantities of biomass, and represents an opportunity to develop, test, and optimize the necessary technologies to make microalgal biofuels more cost-effective and efficient. However, many constraints on the eventual deployment of this technology must be taken into consideration and mitigating strategies developed before large scale microalgal cultivation can become a reality. As a strategy for CO(2) biomitigation from industrial point source emitters, microalgal cultivation can be limited by the availability of land, light, and other nutrients like N and P. Effective removal of N and P from municipal wastewater is limited by the processing capacity of available microalgal cultivation systems. Strategies to mitigate against the constraints are discussed.     

Flow cytometry for the development of biotechnological processes with microalgae

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.     

Microalgae–bacteria symbiosis in microalgal growth and biofuel production: a review

Photosynthetic microalgae can capture solar energy and convert it to bioenergy and biochemical products. In nature or industrial processes, microalgae live together with bacterial communities and may maintain symbiotic relationships. In general interactions, microalgae exude dissolved organic carbon that becomes available to bacteria. In return, the bacteria remineralize sulphur, nitrogen and phosphorous to support the further growth of microalgae. In specific interactions, heterotrophic bacteria supply B vitamins as organic cofactors or produce siderophores to bind iron, which could be utilized by microalgae, while the algae supply fixed carbon to the bacteria in return. In this review, we focus on mutualistic relationship between microalgae and bacteria, summarizing recent studies on the mechanisms involved in microalgae–bacteria symbiosis. Symbiotic bacteria on promoting microalgal growth are described and the relevance of microalgae–bacteria interactions for biofuel production processes is discussed. Symbiotic microalgae–bacteria consortia could be utilized to improve microalgal biomass production and to enrich the biomass with valuable chemical and energy compounds. The suitable control of such biological interactions between microalgae and bacteria will help to improve the microalgae-based biomass and biofuel production in the future.     

Dual purpose microalgae-bacteria-based systems that treat wastewater and produce biodiesel and chemical products within a Biorefinery

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.     

Co-Cultivation of Fungal and Microalgal Cells as an Efficient System for Harvesting Microalgal Cells,Lipid Production and Wastewater Treatment

The challenges which the large scale microalgal industry is facing are associated with the high cost of key operations such as harvesting, nutrient supply and oil extraction. The high-energy input for harvesting makes current commercial microalgal biodiesel production economically unfeasible and can account for up to 50% of the total cost of biofuel production. Co-cultivation of fungal and microalgal cells is getting increasing attention because of high efficiency of bio-flocculation of microalgal cells with no requirement for added chemicals and low energy inputs. Moreover, some fungal and microalgal strains are well known for their exceptional ability to purify wastewater, generating biomass that represents a renewable and sustainable feedstock for biofuel production. We have screened the flocculation efficiency of the filamentous fungus A. fumigatus against 11 microalgae representing freshwater, marine, small (5 µm), large (over 300 µm), heterotrophic, photoautotrophic, motile and non-motile strains. Some of the strains are commercially used for biofuel production. Lipid production and composition were analysed in fungal-algal pellets grown on media containing alternative carbon, nitrogen and phosphorus sources contained in wheat straw and swine wastewater, respectively. Co-cultivation of algae and A. fumigatus cells showed additive and synergistic effects on biomass production, lipid yield and wastewater bioremediation efficiency. Analysis of fungal-algal pellet''s fatty acids composition suggested that it can be tailored and optimised through co-cultivating different algae and fungi without the need for genetic modification.     

Evaluation of electro‐coagulation–flocculation for harvesting marine and freshwater microalgae

Although microalgae are considered as a promising feedstock for biofuels, the energy efficiency of the production process needs to be significantly improved. Due to their small size and low concentration in the culture medium, cost‐efficient harvesting of microalgae is a major challenge. In this study, the use of electro‐coagulation–flocculation (ECF) as a method for harvesting a freshwater (Chlorella vulgaris) and a marine (Phaeodactylum tricornutum) microalgal species is evaluated. ECF was shown to be more efficient using an aluminum anode than using an iron anode. Furthermore, it could be concluded that the efficiency of the ECF process can be substantially improved by reducing the initial pH and by increasing the turbulence in the microalgal suspension. Although higher current densities resulted in a more rapid flocculation of the microalgal suspension, power consumption, expressed per kg of microalgae harvested, and release of aluminum were lower when a lower current density was used. The aluminum content of the harvested microalgal biomass was less than 1% while the aluminum concentration in the process water was below 2 mg L⁻¹. Under optimal conditions, power consumption of the ECF process was around 2 kWh kg⁻¹ of microalgal biomass harvested for Chlorella vulgaris and ca. 0.3 kWh kg⁻¹ for Phaeodactylum tricornutum. Compared to centrifugation, ECF is thus more energy efficient. Because of the lower power consumption of ECF in seawater, ECF is a particularly attractive method for harvesting marine microalgae. Biotechnol. Bioeng. 2011;108: 2320–2329. © 2011 Wiley Periodicals, Inc.