Development of a new method for simultaneous extraction of chlorophylls and carotenoids from microalgal biomass

Journal of Applied Phycology - There are still limitations in the pigment extraction methods used in microalgae biomass, especially for laboratory scale. This work aimed to develop a simple method...     

应用叶绿素荧光法测定微藻生物量的方法

为了探求叶绿素荧光值与海洋微藻生物量的关系,利用TD-700型叶绿素荧光仪测定了6种典型海洋赤潮藻在不同生长期(指数生长期和稳定生长期)时的叶绿素荧光值,同时用传统方法测定细胞密度,且利用Leica Qwin软件测量细胞大小。结果显示:微藻在同一生长时期细胞密度与叶绿素荧光值存在极显著的正相关关系,不同生长时期细胞密度也与单位叶绿素荧光值存在较显著正相关关系。因此,利用叶绿素荧光仪测定微藻生物量的方法是切实可行的,具有快捷方便、灵敏度高、可靠性强的优点。     

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.     

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.     

Recovery of microalgal biomass and metabolites: process options and economics

Commercial production of intracellular microalgal metabolites requires the following: (1) large-scale monoseptic production of the appropriate microalgal biomass; (2) recovery of the biomass from a relatively dilute broth; (3) extraction of the metabolite from the biomass; and (4) purification of the crude extract. This review examines the options available for recovery of the biomass and the intracellular metabolites from the biomass. Economics of monoseptic production of microalgae in photobioreactors and the downstream recovery of metabolites are discussed using eicosapentaenoic acid (EPA) recovery as a representative case study.     

Anaerobic digestion of microalgal biomass with ultrasonic disintegration

The use of photosynthetic microalgae for nutrient removal and biofuel production has been widely discussed. Anaerobic digestion of waste microalgal biomass to produce biogas is a promising technology for bioenergy production. However, the methane yield from this anaerobic process was limited because of the hard cell wall of Chlorella vulgaris. The use of ultrasound has proven to be successful at improving the disintegration and anaerobic biodegradability of Chlorella vulgaris. Ultrasonic pretreatment in the range of 5–200 J ml⁻¹ was applied to waste microalgal biomass, which was then used for batch digestion. Ultrasound techniques were successful and showed higher soluble COD at higher applied energy. During batch digestion, cell disintegration due to ultrasound increased in terms of specific biogas production and the degradation rate. Compared to the untreated sample, the specific biogas production was increased in the ultrasound-treated sample by 90% at an energy dose of 200 J ml⁻¹. For the disintegrated samples, volatile solids reduction was also increased according to the energy input and degradation. These results indicate that the hydrolysis of microalgal cells is the rate-limiting step in the anaerobic digestion of microalgal biomass.     

Interactive biosorption by microalgal biomass as a tool for fluoride removal

Maximum biosorption of Ca²⁺ was at 50 mg Ca²⁺ l^–1 with both Anabaena fertilissima (2.8 mg Ca²⁺ g^–1 dry wt) and Chlorococcum humicola (4.4 mg g^–1). Such Ca²⁺-treated biomasses, accumulated, respectively, 7 mg F g^–1 DW from an aqueous solution of 10 mg F l^–1 and 4.5 mg F g^–1 DW from 15 mg F l^–1. Data for both Ca²⁺ and F^– biosorption fitted the Langmuir adsorption isotherm indicating monolayer adsorption at a constant energy.     

Harnessing energy from plant biomass

Biofuels derived from renewable plant biomass offer a potential carbon-neutral replacement for current liquid transportation fuels. Progress toward this initiative requires development of new methods to engineer energy crops with the desired chemical composition and physical characteristics, depolymerize lignocellulose to fermentable units, and program microbial metabolism for efficient conversion of sugars to ethanol.     

Spouting of biomass particles: a review

Recent research on biomass multiphase flow in spouted beds is reviewed, beginning with fundamental work on hydrodynamic parameters, such as minimum spouting velocity, pressure drop and fountain height. We then consider experimental studies on biomass multiphase flow in such processes as pulp drying, liquid spouting of pulp fibres, drying and coating of agricultural biomass, and bioreactors. Finally, we summarize modelling efforts with respect to spouting of biomass particles.     

基于文献计量学的微藻生物技术发展趋势

微藻作为一类单细胞光合生物,具有光能利用率高、生长速度快、生物活性成分和储能物质含量高等特点,在食品、饲料、生物能源、碳减排和废水处理等领域具有广阔的应用前景。本文系统查阅了近年来国内外微藻领域发表的相关文献和专利,藉此对微藻生物技术领域的研究进展及存在问题进行了梳理和分析,并对我国微藻生物技术的科技布局、重要成果及发展瓶颈进行了概述,最后对微藻生物技术的发展趋势予以展望。     

Methods for DNA extraction from various soils: a comparison

Seven methods for bacterial DNA extraction and purification from soil samples were compared. Holben's direct lysis method recovered significantly greater amounts of DNA than the other methods tested, while CsCl-ethidium bromide density gradient ultracentrifugation was better than gel filtration at removing humic acid from crude DNA isolated from soil. When both these methods were combined, 5·94 μg of DNA (A_260/280 ratio around 1·754) was yielded g⁻¹ oven-dried sandstone shale alluvial soil; similarly satisfactory yields were obtained from Taiwan clay, and sandstone shale and slate alluvial soil managed under different farming practices. DNA obtained by these methods was readily digested by Eco R I and Hind III. When soil samples were stored for 3 weeks at 4 °C, the fraction of high-molecular-weight DNA was reduced significantly. Thus, DNA extraction should be carried out as soon as possible after a soil sample has been collected from the field. When hyphae of Pythium aphanidermatum and Fusarium solani were subjected to the above lysis method, DNA could not be detected in the extract.     

Stylized facts in microalgal growth: interpretation in a dynamic energy budget context

A dynamic energy budget (DEB) model for microalgae is proposed. This model deviates from the standard DEB model as it needs more reserves to cope with the variation of assimilation pathways, requiring a different approach to growth based on the synthesizing unit (SU) theory for multiple substrates. It is shown that the model is able to accurately predict experimental data in constant and light-varying conditions with most of the parameter values taken directly from the literature. Also, model simulations are shown to be consistent with stylized facts (SFs) concerning NC ratio. These SFs are reinterpreted and the general conclusion is that all forcing variables (dilution rate, temperature and irradiance) impose changes in the nitrogen or carbon limitation status of the population, and consequently on reserve densities. Model predictions are also evaluated in comparison with SFs on chlorophyll concentration. It is proposed that an extra structure, more dependent on the nitrogen reserve, is required to accurately model chlorophyll dynamics. Finally, SFs concerning extracellular polymeric substances (EPSs) production by benthic diatoms are collected and interpreted and a formulation based on product synthesis and rejection flux is proposed for the EPSs production rate.     

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.     

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.     

Recycling and reuse of spent microalgal biomass for sustainable biofuels

The use of microalgal biomass (MAB) for biofuel production has been recognized since long. Despite distinct advantages of algal biofuels, however, their sustainability and economic viability is still doubtful. Overall process cost and low energy recovery need to be significantly improved. The use of MAB, after extracting primary fuels in the form of hydrogen, methane, biodiesel and bioethanol, can be one promising route. This algal biomass, collectively termed as spent microalgal biomass (SMAB), contains even up to 70% of its initial energy level and also retains nutrients including proteins, carbohydrates, and lipids. Potential application routes include diet for animals and fish, the removal of heavy metals and dyes from wastewater, and the production of bioenergy (e.g., biofuels and electricity). Unlike whole algae biomass whose applications are relatively well documented, SMAB has been studied only to limited degree. Therefore, this work gives a brief overview of various ways of SMAB applications. An insight into current status, barriers and future prospects on SMAB research is provided. The feasibility of each application is evaluated on the basis of its energy recovery, economic viability, and future perspectives are provided.     

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.     

The potential of microalgal biotechnology: a review of production and uses of microalgae

An overview of the various aspects, promises and limitations of microalgal biotechnology is presented. The factors of importance in microalgal cultivation as well as the culture systems are briefly described. Microalgal biomasses can fulfil the nutritional requirements of aquatic larvae and organisms. The biochemical composition of algae can be improved by the manipulation of culture conditions. The nutritive value of the microalgal biomasses for human and animal consumption is also commented upon as well as some socio-economical aspects. Among the sources of required nutrients (N, P), wastewaters and manures can upgraded as culture media for microalgae the safety of which has to be evaluated. Harvesting of the biomass is one of the bottlenecks. The various techniques, physical, physico-chemical and biological are outlined and their feasibility and economic interest examined. Microalgal biomasses can be submitted to various technological transformations. Various processes are reviewed in the light of their effects on safety and nutritional value. The possible extraction of fine chemicals and the preparation of protein concentrates is also reported on. The various uses of microalgae lead to a possible competition, to be evaluated, between systems for the production of food, energy and chemicals. The review finally covers the application of genetic manipulation to microalgae.     

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.     

Ethanol production from lignocellulosic biomass of energy cane

Ethanol produced from lignocellulosic biomass is a renewable alternative to diminishing petroleum based liquid fuels. The release of many new sugarcane varieties by the United States Department of Agriculture to be used as energy crops is a promising feedstock alternative. Energy cane produces large amounts of biomass that can be easily transported, and production does not compete with food supply and prices because energy cane can be grown on marginal land instead of land for food crops. The purpose of this study was to evaluate energy cane for lignocellulosic ethanol production. Energy cane variety L 79-1002 was pretreated with weak sulfuric acid to remove lignin. In this study, 1.4 M sulfuric acid pretreated type II energy cane had a higher ethanol yield after fermentation by Klebsiella oxytoca without enzymatic saccharification than 0.8 M and 1.6 M sulfuric acid pretreated type II energy cane. Pretreated biomass was inoculated with K. oxytoca for cellulose fermentation and Pichia stipitis for hemicellulose fermentation under simultaneous saccahrification and fermentation (SSF) and separate hydrolysis and fermentation (SHF) conditions. For enzymatic saccharification of cellulose, the cellulase and ??-glucanase cocktail significantly increased ethanol production compared to the ethanol production of fermented acid pretreated energy cane without enzymatic saccharification. The results revealed that energy cane variety L 79-1002 produced maximum cellulosic ethanol under SHF (6995 mg/L) and produced 3624 mg/L ethanol from fermentation of hemicellulosic sugars.