Characterization of the acylglycerols and resulting biodiesel derived from vegetable oil and microalgae (Thalassiosira pseudonana and Phaeodactylum tricornutum)

Algal biofuels are a growing interest worldwide due to their potential in terms of sustainable greenhouse gas displacement and energy production. This article describes a comparative survey of biodiesel production and conversion yields of biodiesel via alkaline transesterification of acylglycerols extracted from the microalgae Thalassiosira pseudonana and Phaeodactylum tricornutum, grown under silicate or nitrate limitation, and that of model vegetable oils: soybean, and rapeseed oil. Acylglycerols were extracted with n-hexane and the total yield per biomass was determined by gravimetric assay. Under our conditions, the total acylglycerol yield from the microalgae studied was 13-18% of total dry weight. The biodiesel samples were analyzed using gas chromatography-flame ionization detector to determine quantitative information of residual glycerol, mono-, di-, and tri-acylglycerol concentrations in the biodiesel. All of the algal-based biodiesel demonstrated less mono-, di-, and tri-acylglycerol concentrations than the vegetable-based biodiesel under identical transesterification conditions. The fatty acid compositions of all the feedstock oils and their resultant biodiesel were also analyzed and reported. Based on the fatty acid methyl ester compositions of our samples we qualitatively assessed the suitability of the algal-derived biodiesel in terms of cetane number (CN), cold-flow properties, and oxidative stability.     

Biocatalytic transesterification of triglycerides and alcohols for the production of biodiesel using cutinase in organic media

Abstract

The world's energy supply is mainly composed of fossil fuels, which are a non-renewable source of energy that is rapidly running out. To overcome this concern, industry has been focusing on the production of biofuels such as biodiesel. A range of approaches has been considered to transform oils into applicable biodiesel: dilutions, microemulsifications, pyrolysis and transesterification. The latter method consists of the conversion of triglycerides to a mixture of alkyl esters and glycerol, in the presence of an acyl acceptor and a catalyst. Due to high selectivity when using enzymes as catalysts, and mild operating conditions, biocatalytic transesterification has proven to be an efficient method. Cutinase, from the superfamily α/β hydrolases, is an enzyme with lipolytic activity that effectively catalyses transesterification reactions. This article highlights the use of cutinase microencapsulated in bis(2ethylhexyl) sodium sulfosuccinate (AOT)-reversed micelles to perform the biocatalytic transesterification of triglycerides, with low chain-length alcohols (e.g. methanol), in organic media to produce biodiesel.     

Biodiesel fuel production via transesterification of oils using lipase biocatalyst

Biodiesel has gained widespread importance in recent years as an alternative, renewable liquid transportation fuel. It is derived from natural triglycerides in the presence of an alcohol and an alkali catalyst via a transesterification reaction. To date, transesterification based on the use of chemical catalysts has been predominant for biodiesel production at the industrial scale due to its high conversion efficiency at reasonable cost. Recently, biocatalytic transesterification has received considerable attention due to its favorable conversion rate and relatively simple downstream processing demands for the recovery of by-products and purification of biodiesel. Biocatalysis of the transesterification reaction using commercially purified lipase represents a major cost constraint. However, more cost-effective techniques based on the immobilization of both extracellular and intracellular lipases on support materials facilitate the reusability of the catalyst. Other variables, including the presence of alcohol, glycerol and the activity of water can profoundly affect lipase activity and stability during the reaction. This review evaluates the current status for lipase biocatalyst-mediated production of biodiesel, and identifies the key parameters affecting lipase activity and stability. Pioneer studies on reactor-based lipase conversion of triglycerides are presented.     

Energy recovery from lipid extracted,transesterified and glycerol codigested microalgae biomass

The production of methane (CH₄) via the anaerobic digestion of microalgae biomass residues from the biodiesel production process has the potential to meet some of the energy requirements of the primary biomass to fuel conversion process. This paper investigates the practical CH₄ yields achievable from the anaerobic conversion of the microalgae residues (as well as codigestion with glycerol) after biodiesel production using both the conventional and in situ transesterification methods. Results demonstrate that the type of lipid extraction solvent utilized in the conventional transesterification process could inhibit subsequent CH₄ production. On the basis of actual CH₄ production, a recoverable energy of 8.7–10.5 MJ kg^?1 of dry microalgae biomass residue was obtained using the lipid extracted and transesterified microalgae samples. On codigesting the microalgae residues with glycerol, a 4–7% increase in CH₄ production was observed.     

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.     

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.     

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.     

微藻生物柴油研发态势分析

微藻是光合效率最高的原始植物之一,与农作物相比,单位面积的产率可高出数十倍。微藻生物柴油技术首先包括微藻的筛选和培育,获得性状优良的高含油量藻种,然后在光生物反应器中吸收阳光、CO2等,生成微藻生物质,最后经过采收、加工,转化为微藻生物柴油。完整的微藻生物柴油成套技术链涵盖多个技术环节,是一个复杂的系统工程,包括微藻生物工程技术、微藻高效规模化养殖技术,以及微藻生物质采收、加工与转化技术等。其中,降低生产成本是当前微藻生物柴油研究面临的主要挑战,各国的研究机构为此开展了多方面的研究。     

An efficient and scalable extraction and quantification method for algal derived biofuel

Microalgae are capable of synthesizing a multitude of compounds including biofuel precursors and other high value products such as omega-3-fatty acids. However, accurate analysis of the specific compounds produced by microalgae is important since slight variations in saturation and carbon chain length can affect the quality, and thus the value, of the end product. We present a method that allows for fast and reliable extraction of lipids and similar compounds from a range of algae, followed by their characterization using gas chromatographic analysis with a focus on biodiesel-relevant compounds. This method determines which range of biologically synthesized compounds is likely responsible for each fatty acid methyl ester (FAME) produced; information that is fundamental for identifying preferred microalgae candidates as a biodiesel source. Traditional methods of analyzing these precursor molecules are time intensive and prone to high degrees of variation between species and experimental conditions. Here we detail a new method which uses microwave energy as a reliable, single-step cell disruption technique to extract lipids from live cultures of microalgae. After extractable lipid characterization (including lipid type (free fatty acids, mono-, di- or tri-acylglycerides) and carbon chain length determination) by GC–FID, the same lipid extracts are transesterified into FAMEs and directly compared to total biodiesel potential by GC–MS. This approach provides insight into the fraction of total FAMEs derived from extractable lipids compared to FAMEs derived from the residual fraction (i.e. membrane bound phospholipids, sterols, etc.). This approach can also indicate which extractable lipid compound, based on chain length and relative abundance, is responsible for each FAME. This method was tested on three species of microalgae; the marine diatom Phaeodactylum tricornutum, the model Chlorophyte Chlamydomonas reinhardtii, and the freshwater green alga Chlorella vulgaris. The method is shown to be robust, highly reproducible, and fast, allowing for multiple samples to be analyzed throughout the time course of culturing, thus providing time-resolved information regarding lipid quantity and quality. Total time from harvesting to obtaining analytical results is less than 2 h.     

Zero-waste biorefinery of oleaginous microalgae as promising sources of biofuels and biochemicals through direct transesterification and acid hydrolysis

Oleaginous microalgae are considered as promising sources of biofuels and biochemicals due to their high lipid content and other high-value components such as pigments, carbohydrate and protein. This study aimed to develop an efficient biorefinery process for utilizing all of the components in oleaginous microalgae. Acetone extraction was used to recover microalgal pigments prior to processes for the other products. Microalgal lipids were converted into biodiesel (fatty acid methyl ester, FAME) through a conventional two-step process of lipid extraction followed by transesterification, and alternatively a one-step direct transesterification. The comparable FAME yields from both methods indicate the effectiveness of direct transesterification. The operating parameters for direct transesterification were optimized through response surface methodology (RSM). The maximum FAME yield of 256 g/kg-biomass was achieved when using chloroform:methanol as co-solvents for extracting and reacting reagents at 1.35:1 volumetric ratio, 70 °C reaction temperature, and 120 min reaction time. The carbohydrate content in lipid-free microalgal biomass residues (LMBRs) was subsequently acid hydrolyzed into sugars under optimized conditions from RSM. The maximum sugar yield obtained was 44.8 g/kg-LMBRs and the protein residues were recovered after hydrolysis. This biorefinery process may contribute greatly to zero-waste industrialization of microalgae based biofuels and biochemicals.     

Isolation of thermo-tolerant and high lipid content green microalgae: oil accumulation is predominantly controlled by photosystem efficiency during stress treatments in Desmodesmus

Discoveries of new microalgae with thermo-tolerance, high growth rate, and high lipid content are crucial to algal biodiesel production in tropical and subtropical zones. Four new green microalgae were isolated in southern Taiwan. All four species are members of the genus Desmodesmus under the family Scenedesmaceae based on molecular and morphological analyses. Two of the four species survived at 45 °C for 24 h, with 5-13% of mortality rates caused by the heat. Total lipid contents of the two species reached over 50% in dry biomass under nitrogen starvation, and their triacylglycerols constituted around 75% of the total lipids. Thus the two species are good potential feedstocks for biodiesel production. Oil accumulation in the four species positively correlates with their photosystem II efficiencies during stress treatments (R2=0.90). This finding further supports that photosynthesis is essential for oil body formation under nitrogen starvation in green microalgae.     

Production of Biodiesel Using Immobilized Lipase—A Critical Review

Increase in volume of biodiesel production in the world scenario proves that biodiesel is accepted as an alternative to conventional fuel. Production of biodiesel using alkaline catalyst has been commercially implemented due to its high conversion and low production time. For the product and process development of biodiesel, enzymatic transesterification has been suggested to produce a high purity product with an economic, environment friendly process at mild reaction conditions. The enzyme cost being the main hurdle can be overcome by immobilization. Immobilized enzyme, which has been successfully used in various fields over the soluble counterpart, could be employed in biodiesel production with the aim of reducing the production cost by reusing the enzyme. This review attempts to provide an updated compilation of the studies reported on biodiesel production by using lipase immobilized through various techniques and the parameters, which affect their functionality.     

Effects of outdoor cultures on the growth and lipid production of <Emphasis Type="Italic">Phaeodactylum tricornutum</Emphasis> using closed photobioreactors

One of the principal challenges for large scale production of microalgae is the high costs of biomass production. Aiming for minimize this problem, microalgal biodiesel production should focus on outdoors cultures, using available solar light and allowing lower energy cost process. Testing species that proved to be common and easy to culture may be a good approach in this process. The present work reports indoor-outdoor cultures of Phaeodactylum tricornutum using different bioreactors types, using cell growth, biochemical composition, and the profiles of the fatty acids produced as the parameters to test the optimization processes. The results show that the use of outdoor cultures is a good choice to obtain P. tricornutum biomass with a good potential for biodiesel production. The microalgae produced reached better growth efficiency, major lipid content and showed an increment in the percentage of saturated fatty acids (required on the biodiesel production) respect indoor cultures. These results are important to show the relevance of using outdoor cultures as a way to improve the efficiency and the energetic balance of the biodiesel production with P. tricornutum algae.     

Isolation and evaluation of oil-producing microalgae from subtropical coastal and brackish waters

Microalgae have been widely reported as a promising source of biofuels, mainly based on their high areal productivity of biomass and lipids as triacylglycerides and the possibility for cultivation on non-arable land. The isolation and selection of suitable strains that are robust and display high growth and lipid accumulation rates is an important prerequisite for their successful cultivation as a bioenergy source, a process that can be compared to the initial selection and domestication of agricultural crops. We developed standard protocols for the isolation and cultivation for a range of marine and brackish microalgae. By comparing growth rates and lipid productivity, we assessed the potential of subtropical coastal and brackish microalgae for the production of biodiesel and other oil-based bioproducts. This study identified Nannochloropsis sp., Dunaniella salina and new isolates of Chlorella sp. and Tetraselmis sp. as suitable candidates for a multiple-product algae crop. We conclude that subtropical coastal microalgae display a variety of fatty acid profiles that offer a wide scope for several oil-based bioproducts, including biodiesel and omega-3 fatty acids. A biorefinery approach for microalgae would make economical production more feasible but challenges remain for efficient harvesting and extraction processes for some species.     

A meta-analysis of the life cycle greenhouse gas balances of microalgae biodiesel

Purpose

Microalgae biodiesel has attracted considerable attention as a potential substitute for fossil fuels and biodiesel from food crops. Nevertheless, its reported climate impacts in the scientific literature vary significantly. This article describes and synthesizes the range of results found in the life cycle assessment (LCA) literature regarding microalgae biodiesel studies to investigate whether particular parameters, e.g. technologies, were associated with higher or lower greenhouse gas (GHG) emissions so that a best practice can be inferred from currently available LCA data and thereby recommended.

Methods

A systematic literature review and meta-regression analysis (MRA) of 36 LCA studies that report on the GHG emissions of microalgae biodiesel was conducted. An assessment of key aspects, including modelling choices and technologies, was performed. Furthermore, MRA models were formulated considering several variables of interest describing both technical and modelling choices to identify the main causes for the variability in GHG emissions per MJ of biodiesel. Variables chosen include: microalgae species; culture medium; cultivation system; source of CO₂; extraction technology; conversion technology; system boundary; geographical scope; inclusion or exclusion of capital goods; and how multifunctionality was handled.

Results and discussion

The reviewed studies altogether reported 308 results ranging from ?0.7 to 3.8 kg CO₂ eq. MJ^?1_biodiesel, portraying 19 different system configurations. Despite the comprehensive range of variables assessed, the models generated could not plausibly explain that the variability in GHG emissions depends either on the technologies considered or on the methodological choices adopted. However, the following relationships could be observed: location in Europe and high oil productivity were associated with lower emissions, whilst dry extraction should be avoided for leading to higher GHG emissions, on average.

Conclusions

There is a large degree of variability within the technologies considered, as well as the methodological choices adopted, so that no robust conclusions could be drawn from the MRA. Notwithstanding, average GHG emissions reported were more than twice as high as fossil diesel and, while there are some studies showing large benefits, none of the various algae technologies performed consistently better than fossil diesel, questioning the climate-mitigation potential of microalgae biodiesel.

     

微藻油脂合成代谢途径及调控机制的研究进展

随着经济的快速发展,各国对石油的需求仍有增无减,但是石油作为不可再生能源不利于可持续发展。相比之下,生物柴油应运而生且已发展到了第三代,而微藻作为第三代生物柴油的主角因为具有生长速度快,不占用耕地等优势,已逐渐成为具有极大发展潜力的能源原料,所以利用微藻生产生物柴油的技术近年来也成为研究的重点。本综述针对微藻容易进行基因改造的优势,综述了国内外对微藻油脂代谢通路的研究现状及进展,讨论了目前的基因改造方法对通路中关键酶产生的影响,以求找到能有效并稳定增强微藻油脂代谢途径的方法,为以后的研究提供理论指导。     

Biodiesel production from Jatropha curcas: a critical review

The fuel crisis and environmental concerns, mainly due to global warming, have led researchers to consider the importance of biofuels such as biodiesel. Vegetable oils, which are too viscous to be used directly in engines, are converted into their corresponding methyl or ethyl esters by a process called transesterification. With the recent debates on “food versus fuel,” non-edible oils, such as Jatropha curcas, are emerging as one of the main contenders for biodiesel production. Much research is still needed to explore and realize the full potential of a green fuel from J. curcas. Upcoming projects and plantations of Jatropha in countries such as India, Malaysia, and Indonesia suggest a promising future for this plant as a potential biodiesel feedstock. Many of the drawbacks associated with chemical catalysts can be overcome by using lipases for enzymatic transesterification. The high cost of lipases can be overcome, to a certain extent, by immobilization techniques. This article reviews the importance of the J. curcas plant and describes existing research conducted on Jatropha biodiesel production. The article highlights areas where further research is required and relevance of designing an immobilized lipase for biodiesel production is discussed.     

我国微藻生物柴油的研究背景与发展战略

生物柴油是可再生能源开发利用的重要发展方向。藻类制备生物柴油具有产油量高、生长速度快、环境适应能力强、不与农作物争夺农田和淡水资源等优势。从微藻制备生物柴油着手,简要介绍了生物柴油的生产原料与发展历程、微藻油脂的组成与生物合成途径、微藻制备柴油的工艺与瓶颈及解决策略,最后对微藻制备生物柴油技术提出了近中远期发展目标及展望。     

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.     

Life-cycle assessment of microalgae culture coupled to biogas production

Due to resource depletion and climate change, lipid-based algal biofuel has been pointed out as an interesting alternative because of the high productivity of algae per hectare and per year and its ability to recycle CO₂ from flue gas. Another option for taking advantage of the energy content of the microalgae is to directly carry out anaerobic digestion of raw algae in order to produce methane and recycle nutrients (N, P and K). In this study, a life-cycle assessment (LCA) of biogas production from the microalgae Chlorella vulgaris is performed and the results are compared to algal biodiesel and to first generation biodiesels. These results suggest that the impacts generated by the production of methane from microalgae are strongly correlated with the electric consumption. Progresses can be achieved by decreasing the mixing costs and circulation between different production steps, or by improving the efficiency of the anaerobic process under controlled conditions. This new bioenergy generating process strongly competes with others biofuel productions.