Engineered chlorophyll catabolism conferring predator resistance for microalgal biomass production

Production of valuable compounds including biofuels and pharmaceutical precursors derived from microalgae has garnered significant interest. Stable production of algal biomass is essential to make the microalgal industry commercially feasible. However, one of the largest issues is severe biological contamination by predators grazing the algal biomass, resulting in the crash of outdoor cultures. In the present study, we propose a novel engineering strategy for microalgae to cope with predators. The overexpression of plant chlorophyllase (CLH) in a microalga resulted in the enhancement of resistance to the predator. This result supported our hypothesis that CLH promotes chlorophyll breakdown in the chloroplasts of the microalgae when they are digested by the predator, generating the phototoxic catabolite chlorophyllide that damages the predator. To the best of our knowledge, this is the first study to establish predator-resistant microalgae by enhancing the CLH activity.     

Microalgal triacylglycerols as feedstocks for biofuel production: perspectives and advances

Microalgae represent an exceptionally diverse but highly specialized group of micro-organisms adapted to various ecological habitats. Many microalgae have the ability to produce substantial amounts (e.g. 20–50% dry cell weight) of triacylglycerols (TAG) as a storage lipid under photo-oxidative stress or other adverse environmental conditions. Fatty acids, the building blocks for TAGs and all other cellular lipids, are synthesized in the chloroplast using a single set of enzymes, of which acetyl CoA carboxylase (ACCase) is key in regulating fatty acid synthesis rates. However, the expression of genes involved in fatty acid synthesis is poorly understood in microalgae. Synthesis and sequestration of TAG into cytosolic lipid bodies appear to be a protective mechanism by which algal cells cope with stress conditions, but little is known about regulation of TAG formation at the molecular and cellular level. While the concept of using microalgae as an alternative and renewable source of lipid-rich biomass feedstock for biofuels has been explored over the past few decades, a scalable, commercially viable system has yet to emerge. Today, the production of algal oil is primarily confined to high-value specialty oils with nutritional value, rather than commodity oils for biofuel. This review provides a brief summary of the current knowledge on oleaginous algae and their fatty acid and TAG biosynthesis, algal model systems and genomic approaches to a better understanding of TAG production, and a historical perspective and path forward for microalgae-based biofuel research and commercialization.     

Microalgae biofuel potentials (Review)

With the decrease of fossil based fuels and the environmental impact of them over the planet, it seems necessary to seek the sustainable sources of clean energy. Biofuels, is becoming a worldwide leader in the development of renewable energy resources. It is worthwhile to say that algal biofuel production is thought to help stabilize the concentration of carbon dioxide in the atmosphere and decrease global warming impacts. Also, among algal fuels’ attractive characteristics, algal biodiesel is non toxic, with no sulfur, highly biodegradable and relatively harmless to the environment if spilled. Algae are capable of producing in excess of 30 times more oil per acre than corn and soybean crops. Currently, algal biofuel production has not been commercialized due to high costs associated with production, harvesting and oil extraction but the technology is progressing. Extensive research was conducted to determine the utilization of microalgae as an energy source and make algae oil production commercially viable.     

Microalgae biofuel potentials (review)

With the decrease of fossil based fuels and the environmental impact of them over the planet, it seems necessary to seek the sustainable sources of clean energy. Biofuels, is becoming a worldwide leader in the development of renewable energy resources. It is worthwhile to say that algal biofuel production is thought to help stabilize the concentration of carbon dioxide in the atmosphere and decrease global warming impacts. Also, among algal fuels' attractive characteristics, algal biodiesel is non toxic, with no sulfur, highly biodegradable and relatively harmless to the environment if spilled. Algae are capable of producing in excess of 30 times more oil per acre than corn and soybean crops. Currently, algal biofuel production has not been commercialized due to high costs associated with production, harvesting and oil extraction but the technology is progressing. Extensive research was conducted to determine the utilization of microalgae as an energy source and make algae oil production commercially viable.     

Microalgae as bioreactors

Microalgae already serve as a major natural source of valuable macromolecules including carotenoids, long-chain polyunsaturated fatty acids and phycocolloids. As photoautotrophs, their simple growth requirements make these primitive plants potentially attractive bioreactor systems for the production of high-value heterologous proteins. The difficulty of producing stable transformants has meant that the field of transgenic microalgae is still in its infancy. Nonetheless, several species can now be routinely transformed and algal biotechnology companies have begun to explore the possibilities of synthesizing recombinant therapeutic proteins in microalgae and the engineering of metabolic pathways to produce increased levels of desirable compounds. In this review, we compare the current commercially viable bioreactor systems, outline recent progress in microalgal biotechnology and transformation, and discuss the potential of microalgae as bioreactors for the production of heterologous proteins.     

High-value products from microalgae—their development and commercialisation

Microalgae (including the cyanobacteria) are established commercial sources of high-value chemicals such as β-carotene, astaxanthin, docosahexaenoic acid, eicosahexaenoic acid, phycobilin pigments and algal extracts for use in cosmetics. Microalgae are also increasingly playing a role in cosmaceuticals, nutraceuticals and functional foods. In the last few years, there has been renewed interest in microalgae as commercial sources of these and other high-value compounds, driven in part by the attempts to develop commercially viable biofuels from microalgae. This paper briefly reviews the main existing and potential high-value products which can be derived from microalgae and considers their commercial development with a particular focus on the various aspects which need to be considered on the path to commercialisation, using the experience gained in the commercialisation of existing algae products. These considerations include the existing and potential market size and market characteristics of the product, competition by chemically synthesised products or by ‘natural’ compounds from other organisms such as fungi, bacteria, higher plants, etc., product quality requirements and assurance, and the legal and regulatory environment.     

Algal biotechnology products and processes — matching science and economics

Several microalgae, such as species ofChlorella, Spirulina andDunaliella, are grown commercially and algal products such as -carotene and phycocyanin are available. The main focus of algal biotechnology continues to be on high value fine chemicals and on algae for use as aquaculture feeds. This paper provides the outline for a rational approach in evaluating which algae and which algal products are the most likely to be commercially viable. This approach involves some simple market analysis followed by economic modelling of the whole production process. It also permits an evaluation of which steps in the production process have the greatest effect on the final production cost of the alga or algal product, thus providing a guide as to what area the research and development effort should be directed to. An example of this approach is presented and compared with other models. The base model used here gives a production cost of microalgal biomass at about AS 14 to 15 kg^–1, excluding the costs of further processing, packaging and marketing. The model also shows that some of the key factors in microalgal production are productivity, labor costs and harvesting costs. Given the existing technology, high value products such as carotenoids and algal biomass for aquaculture feeds have the greatest commercial potential in the short term.This paper was presented at the Symposium on Applied Phycology at the Fourth International Phycological Congress, Duke University.     

Review of advances in biological CO<Subscript>2</Subscript> mitigation technology

CO₂ fixation by microalgae has emerged as a promising option for CO₂ mitigation. Intensive research work has been carried out to develop a feasible system for removing CO₂ from industrial exhaust gases. However, there are still several challenging points to overcome in order to make the process more practical. In this paper, recent research activities on three key technologies of biological CO₂ fixation, an identification of a suitable algal strain, development of high efficient photobioreactor and utilization of algal cells produced, are described. Finally the barriers, progress, and prospects of commercially developing a biological CO₂ fixation process are summarized.     

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.     

Bioflocculation as an innovative harvesting strategy for microalgae

Microalgae are a promising new source of biomass for the production of third generation biofuels but, so far, the majority of microalgal biomass has been used for high-value applications. New low-cost technologies are needed to make the production and processing of microalgae economically feasible for low-value applications. A major challenge lies in the harvesting of microalgae, which requires a cost-efficient separation technology. Flocculation, especially bioflocculation, is an attractive low-cost separation technology. Various new bioflocculation strategies have been claimed to generate major advances in cost-efficient harvesting. Here, we review the recent advances in bioflocculation based on algal–bacterial, algal–fungal, or algal–algal interactions within the framework of microalgae biomass harvesting for biofuel production. We also discuss recent advances using infochemicals and genetic engineering for the induction of bioflocculation.     

Bioprocess engineering of microalgae to optimize lipid production through nutrient management

Microalgae have been used commercially as a feedstock for the production of high-value compounds, pigments, cosmetics, and nutritional supplements. In addition, because of their rapid growth rates, high photosynthetic efficiency, and high lipid and protein content, commodity products including biodiesel, feed supplements, and polyunsaturated fatty acids derived from algal biomass are of current interest. Since microalgae lack non-photosynthetic structures and float in water, they do not need massive amounts of structural cellulose found in land plants. Thus, under optimal culture conditions, some oleaginous species can allocate up to 70 % of their biomass to lipids. Lipid production and its regulation in microalgae are species-specific and influenced by environmental conditions. Various strategies have been developed to improve lipid productivity and fatty acid composition to meet specific production goals. Manipulation of physiochemical parameters, trophic modes, and nutrient levels, known as process engineering, is a simple approach that leads to desired alterations in the biochemical composition of algal biomass, including lipid quantity and quality. In this paper, we review the effects of manipulating biochemical parameters such as necessary nutrients (C, N, P, S, Fe, and Si), NaCl concentration, and pH of culture medium to optimize lipid content and profile in some algae strains with commercial potential.     

Cell fragility — The key problem of microalgae mass production in closed photobioreactors

The gap between the theoretical biological potential of microalgae and the biomass productivity obtained with algal culture in tubular biophotoreactors is due to a reduced growth rate related to hydrodynamic stress of pumping. High levels of mixing are necessary to reach a turbulent flow of the culture, in order to optimize the light regime. The optimal conditions of pumping to produce this significant liquid mixing may produce some cell damage. Factors affecting this hydrodynamic stress (geometry of the bioreactor involved, type of pump utilized, morphology of algal cells, physiological conditions of microalgae, etc.) are discussed.     

Effects of bacterial communities on biofuel-producing microalgae: stimulation,inhibition and harvesting

Despite the great interest in microalgae as a potential source of biofuel to substitute for fossil fuels, little information is available on the effects of bacterial symbionts in mass algal cultivation systems. The bacterial communities associated with microalgae are a crucial factor in the process of microalgal biomass and lipid production and may stimulate or inhibit growth of biofuel-producing microalgae. In addition, we discuss here the potential use of bacteria to harvest biofuel-producing microalgae. We propose that aggregation of microalgae by bacteria to achieve >90% reductions in volume followed by centrifugation could be an economic approach for harvesting of biofuel-producing microalgae. Our aims in this review are to promote understanding of the effects of bacterial communities on microalgae and draw attention to the importance of this topic in the microalgal biofuel field.     

Haemagglutinating and antibiotic activities of freshwater microalgae

We analysed the haemagglutinating activity of algal extracts from 44 species of freshwater microalgae against native and trypsin/papain-treated cow, pig, sheep, and human A-, B-, and O-type erythrocytes. Algal extracts obtained with aqueous ethanol exhibited higher haemagglutinating activity than those obtained with aqueous acetone. Most of the algal extracts agglutinated at least one of the erythrocyte types analysed. Human erythrocytes were the most sensitive of the cell types analysed. In the other species, the sensitivity of algal haemagglutinating activity for erythrocytes was pig > sheep > cow. Pre-treating erythrocytes with trypsin and papain improved the detection of most algal agglutinins and increased the haemagglutination titre; pre-treatment with papain was most effective for pig erythrocytes. Algal extracts stored at –20 °C for 4 months lost their haemagglutinating activity. Algal extracts also exhibited strong antibiotic activity against food pathogenic bacteria, especially against Bacillus. Our numerical taxonomy data showed that these microalgae might be grouped into several clusters according to their haemagglutinating activity. The detection of haemagglutinating activity may provide an efficient biochemical or physiological character to classify and differentiate microalgae. Our results suggest that freshwater microalgae might provide a potent source of haemagglutinins and antibacterial compounds for biochemical and medical studies and applications.     

Mild disintegration methods of microalgae–bacteria flocs from wastewater treatment

The use of microalgae in biological wastewater treatment has been widely studied. However, there is a dearth of information about estimating the microalgae and bacteria concentrations. In order to maintain a stable algal-bacterial system, it is necessary to quantify both the algal and bacterial biomasses. Typically, microalgae and bacteria from flocs in activated sludge contribute to better biomass settleability. However, flocs cause problems when it comes to estimating the individual biomass concentrations of microalgae and bacteria in a symbiotic algae-bacteria aggregate. This study aimed to find the best disintegration treatment with low influence on the viability of the microalgal cell determined by its photosynthetic activity. In the present work, biological (enzyme solution), chemical (formaldehyde), mechanical (glass bead-beating), and physical (sonication) treatments were performed on microalgae–bacteria flocs (ALBA flocs) to disintegrate the community as a pre-treatment step in order to develop a method for estimating the algal and bacterial concentration and to quantify the degree of disintegration. The effectiveness of the methods to disintegrate ALBA flocs in descending order are the following: sonication, bead-beating, formaldehyde and enzyme application. Sonication treatment (40 W, 6 min) showed the best disintegration performance of the microalgal-bacterial flocs, up to 90 % with 17 % loss of the algal photosynthetic activity. Bead-beating (3 mm diameter, 80 s) achieved 80 % of disintegration with only 6 % loss of its photosynthetic activity. These results demonstrate the possibility of mild disintegration of compact ALBA flocs without having any adverse impact on the microalgae cell. After these treatments, it becomes possible to estimate the individual biomass concentrations of algae and bacteria manually such as with a cell-counting chamber.     

有机磷农药对微藻的毒性作用研究概述

论文就有机磷农药对藻类生长的影响、毒性机理以及对浮游植物群落结构的影响进行综述。相对于水生甲壳类和鱼类,有机磷农药对藻类毒性较低,EC₅₀一般高于1mg/L,对藻类生长的影响大致呈现低浓度促进、高浓度抑制的趋势,有机磷农药之间及其与其它化合物之间具有联合毒性作用。有机磷农药对藻细胞酶活性具有一定影响,而对光合作用的影响也是有机磷农药对藻类毒性效应的重要致毒机制。浮游植物对有机磷农药敏感性差异以及施药引起的浮游动物对藻类的选择性摄食,可能会导致水生态系统中浮游植物群落结构的变化。     

Microalgae aquaculture feeds

Microalgae feeds are currently used in relatively small amounts in aquaculture, mainly for the production of larvae and juvenile shell- and finfish, as well as for raising the zooplankton required for feeding of juvenile animals. The blue-green algaSpirulina is used in substantial amounts (over 100 t y^–1) as a fish and shrimp feed, and even larger markets can be projected if production costs could be reduced. Another potential large-scale application of microalgae is the cultivation ofHaematococcus for the production of the carotenoid astaxanthin, which gives salmon flesh its reddish color. In the long-term microalgae biomass high in lipids (omega-3 fatty acids) may be developed as substitutes for fish oil-based aquaculture feeds. In shrimp ponds the indigenous algal blooms supply a part of the dietary requirements of the animals, but it is difficult to maximize algal productivities. A separate algal production system could feed the shrimps and minimize the need for added feed. Bivalves feed essentially exclusively on marine microalgae throughout their life cycle. The development of cultivation technologies for such microalgae would allow the onshore production of these animals, with greatly improved product quality and safety.This paper was presented at the Symposium on Applied Phycology at the Fourth International Phycological Congress, Duke University.     

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.     

A green light for engineered algae: redirecting metabolism to fuel a biotechnology revolution

Microalgae have the potential to revolutionize biotechnology in a number of areas including nutrition, aquaculture, pharmaceuticals, and biofuels. Although algae have been commercially cultivated for over 50 years, metabolic engineering now seems necessary in order to achieve their full processing capabilities. Recently, the development of a number of transgenic algal strains boasting recombinant protein expression, engineered photosynthesis, and enhanced metabolism encourage the prospects of designer microalgae. Given the vast contributions that these solar-powered, carbon dioxide-sequestering organisms can provide to current global markets and the environment, an intensified focus on microalgal biotechnology is warranted. Ongoing advances in cultivation techniques coupled with genetic manipulation of crucial metabolic networks will further promote microalgae as an attractive platform for the production of numerous high-value compounds.     

Axenic cultures for microalgal biotechnology: Establishment,assessment, maintenance,and applications

The impact of microalgae (including blue-green algae or cyanobacteria) on human life can be both beneficiary and deleterious. While microalgae can be cultivated and used as feedstocks for the production of bioenergy and high value-added products in nutraceuticals, pharmaceuticals, and aquaculture feeds, some microalgae cause harmful algal blooms (HABs) that cause large-scale mortality in aquatic environments around the world. Thus, with the development of microalgal biotechnology and increasing concern about HABs, research on microscopic algae has increased significantly. However, this growth of academic research and application fields has been hindered by difficulties in obtaining axenic cultures. Therefore, this review provides a brief explanation of diverse establishment techniques, along with their strengths and weaknesses, with the hope of facilitating successful axenic cultures. A compilation of research fields and relevant important findings is also presented to clarify the importance of pure algal cultures. Finally, several controversial and sometimes overlooked issues related to the establishment, maintenance, and utilization of axenic cultures are discussed.