Biofortification of safflower: an oil seed crop engineered for ALA-targeting better sustainability and plant based omega-3 fatty acids

Alpha-linolenic acid (ALA) deficiency and a skewed n6:n3 fatty acid ratio in the diet is a major explanation for the prevalence of cardiovascular diseases and inflammatory/autoimmune diseases. There is mounting evidence of the health benefits associated with omega-3 long chain polyunsaturated fatty acids (LC PUFA’s). Although present in abundance in fish, a number of factors limit our consumption of fish based omega-3 PUFA’s. To name a few, overexploitation of wild fish stocks has reduced their sustainability due to increased demand of aquaculture for fish oil and meal; the pollution of marine food webs has raised concerns over the ingestion of toxic substances such as heavy metals and dioxins; vegetarians do not consider fish-based sources for supplemental nutrition. Thus alternative sources are being sought and one approach to the sustainable supply of LC-PUFAs is the metabolic engineering of transgenic plants with the capacity to synthesize n3 LC-PUFAs. The present investigation was carried out with the goal of developing transgenic safflower capable of producing pharmaceutically important alpha-linolenic acid (ALA, C18:3, n3). This crop was selected as the seeds accumulate ~?78% of the total fatty acids as linoleic acid (LA, C18:2, n6), the immediate precursor of ALA. In the present work, ALA production was achieved successfully in safflower seeds by transforming safflower hypocotyls with Arabidopsis specific delta 15 desaturase (FAD3) driven by truncated seed specific promoter. Transgenic safflower fortified with ALA is not only potentially valuable nutritional superior novel oil but also has reduced ratio of LA to ALA which is required for good health.     

Heterotrophic production of eicosapentaenoic acid by microalgae

Eicosapentaenoic acid (EPA) is an omega-3 polyunsaturated fatty acid that plays an important role in the regulation of biological functions and prevention and treatment of a number of human diseases such as heart and inflammatory diseases. As fish oil fails to meet the increasing demand for purified EPA, alternative sources are being sought. Microalgae contain large quantities of high-quality EPA and they are considered a potential source of this important fatty acid. Some microalgae can be grown heterotrophically on cheap organic substrate without light. This mode of cultivation can be well controlled and provides the possibility to maximize EPA production on a large scale. Numerous strategies have been investigated for commercial production of EPA by microalgae. These include screening of high EPA-yielding microalgal strains, improvement of strains by genetic manipulation, optimization of culture conditions, and development of efficient cultivation systems. This paper reviews recent advances in heterotrophic production of EPA by microalgae with an emphasis on the use of diatoms as producing organisms.     

Comparative life cycle assessment of rapeseed oil and palm oil

Background, aim and scope  

The environmental effect of globalisation has been debated intensively in the last decades. Only few well-documented analyses of global versus local product alternatives exist, whilst recommendations on buying local are vast. At the same time, the European Environmental Agency’s Third Assessment concludes that the resource use within the EU is stabilising at the expense of increased resource use for import of products to the EU. Taking its point of departure in vegetable oils, this article compares rapeseed oil and palm oil as a local and a global alternative for meeting the increasing demand for these products in the EU. By using detailed life cycle assessment (LCA), this study compares the environmental impacts and identifies alternative ways of producing rapeseed oil and palm oil to the EU market in order to reduce environmental impacts.     

The synthesis and accumulation of stearidonic acid in transgenic plants: a novel source of 'heart-healthy' omega-3 fatty acids

Dietary omega-3 polyunsaturated fatty acids have a proven role in reducing the risk of cardiovascular disease and precursor disease states such as metabolic syndrome. Although most studies have focussed on the predominant omega-3 fatty acids found in fish oils (eicosapentaenoic acid and docosahexaenoic acid), recent evidence suggests similar health benefits from their common precursor, stearidonic acid. Stearidonic acid is a Δ6-unsaturated C18 omega-3 fatty acid present in a few plant species (mainly the Boraginaceae and Primulaceae ) reflecting the general absence of Δ6-desaturation from higher plants. Using a Δ6-desaturase from Primula vialii , we generated transgenic Arabidopsis and linseed lines accumulating stearidonic acid in their seed lipids. Significantly, the P. vialii Δ6-desaturase specifically only utilises α-linolenic acid as a substrate, resulting in the accumulation of stearidonic acid but not omega-6 γ-linolenic acid. Detailed lipid analysis revealed the accumulation of stearidonic acid in neutral lipids such as triacylglycerol but an absence from the acyl-CoA pool. In the case of linseed, the achieved levels of stearidonic acid (13.4% of triacylglycerols) are very similar to those found in the sole natural commercial plant source ( Echium spp.) or transgenic soybean oil. However, both those latter oils contain γ-linolenic acid, which is not normally present in fish oils and considered undesirable for heart-healthy applications. By contrast, the stearidonic acid-enriched linseed oil is essentially devoid of this fatty acid. Moreover, the overall omega-3/omega-6 ratio for this modified linseed oil is also significantly higher. Thus, this nutritionally enhanced linseed oil may have superior health-beneficial properties.     

Biodiesel production from Jatropha oil by catalytic and non-catalytic approaches: an overview

Biodiesel (fatty acids alkyl esters) is a promising alternative fuel to replace petroleum-based diesel that is obtained from renewable sources such as vegetable oil, animal fat and waste cooking oil. Vegetable oils are more suitable source for biodiesel production compared to animal fats and waste cooking since they are renewable in nature. However, there is a concern that biodiesel production from vegetable oil would disturb the food market. Oil from Jatropha curcas is an acceptable choice for biodiesel production because it is non-edible and can be easily grown in a harsh environment. Moreover, alkyl esters of jatropha oil meet the standard of biodiesel in many countries. Thus, the present paper provides a review on the transesterification methods for biodiesel production using jatropha oil as feedstock.     

LCA of soybean meal

Background, Aim and Scope  Soybean meal is an important protein input to the European livestock production, with Argentina being an important supplier. The area cultivated with soybeans is still increasing globally, and so are the number of LCAs where the production of soybean meal forms part of the product chain. In recent years there has been increasing focus on how soybean production affects the environment. The purpose of the study was to estimate the environmental consequences of soybean meal consumption using a consequential LCA approach. The functional unit is ‘one kg of soybean meal produced in Argentina and delivered to Rotterdam Harbor’. Materials and Methods  Soybean meal has the co-product soybean oil. In this study, the consequential LCA method was applied, and co-product allocation was thereby avoided through system expansion. In this context, system expansion implies that the inputs and outputs are entirely ascribed to soybean meal, and the product system is subsequently expanded to include the avoided production of palm oil. Presently, the marginal vegetable oil on the world market is palm oil but, to be prepared for fluctuations in market demands, an alternative product system with rapeseed oil as the marginal vegetable oil has been established. EDIP97 (updated version 2.3) was used for LCIA and the following impact categories were included: Global warming, eutrophication, acidification, ozone depletion and photochemical smog. Results  Two soybean loops were established to demonstrate how an increased demand for soybean meal affects the palm oil and rapeseed oil production, respectively. The characterized results from LCA on soybean meal (with palm oil as marginal oil) were 721 gCO₂ eq. for global warming potential, 0.3 mg CFC11 eq. for ozone depletion potential, 3.1 g SO₂ eq. for acidification potential, −2 g NO₃ eq. for eutrophication potential and 0.4 g ethene eq. for photochemical smog potential per kg soybean meal. The average area per kg soybean meal consumed was 3.6 m²year. Attributional results, calculated by economic and mass allocation, are also presented. Normalised results show that the most dominating impact categories were: global warming, eutrophication and acidification. The ‘hot spot’ in relation to global warming, was ‘soybean cultivation’, dominated by N₂O emissions from degradation of crop residues (e.g., straw) and during biological nitrogen fixation. In relation to eutrophication and acidification, the transport of soybeans by truck is important, and sensitivity analyses showed that the acidification potential is very sensitive to the increased transport distance by truck. Discussion  The potential environmental impacts (except photochemical smog) were lower when using rapeseed oil as the marginal vegetable oil, because the avoided production of rapeseed contributes more negatively compared with the avoided production of palm oil. Identification of the marginal vegetable oil (palm oil or rapeseed oil) turned out to be important for the result, and this shows how crucial it is in consequential LCA to identify the right marginal product system (e.g., marginal vegetable oil). Conclusions  Consequential LCAs were successfully performed on soybean meal and LCA data on soybean meal are now available for consequential (or attributional) LCAs on livestock products. The study clearly shows that consequential LCAs are quite easy to handle, even though it has been necessary to include production of palm oil, rapeseed and spring barley, as these production systems are affected by the soybean oil co-product. Recommendations and Perspectives  We would appreciate it if the International Journal of Life Cycle Assessment had articles on the developments on, for example, marginal protein, marginal vegetable oil, marginal electricity (related to relevant markets), marginal heat, marginal cereals and, likewise, on metals and other basic commodities. This will not only facilitate the work with consequential LCAs, but will also increase the quality of LCAs.     

Sustainable production of eicosapentaenoic acid-rich oil from microalgae: Towards an algal biorefinery

Utilization of sustainable natural resources such as microalgae has been considered for the production of biofuels, aquaculture feed, high-value bioactives such as omega-3 fatty acids, carotenoids, etc. Eicosapentaenoic acid (EPA) is an omega-3 fatty acid present in fish oil, which is of physiological importance to both humans and fish. Marine microalgae are sustainable sources of lipid rich in EPA, and different species have been explored for the production of EPA as a single product. There has been a rising interest in the concept of a multi-product biorefinery, focusing on the maximum valorization of the algal biomass. Targeting one or more value-added compounds in a biorefinery scenario can improve the commercial viability of low-value products such as triglycerides for biofuel. This approach has been viewed by technologists and experts as a sustainable and economically feasible possibility for the large-scale production of microalgae for its potential applications in biodiesel and jet fuel production, nutraceuticals, animal and aquaculture feeds, etc. In this review paper, we describe the recent developments in the production of high-value EPA-rich oil from microalgae, emphasizing the upstream and downstream bioprocess techniques, and the advantages of considering an EPA-rich oil-based biorefinery.     

Microbial and genetically engineered oils as replacements for fish oil in aquaculture feeds

As the global population grows more of our fish and seafood are being farmed. Fish are the main dietary source of the omega-3 (n-3) long-chain polyunsaturated fatty acids (LC-PUFA), eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids, but these cannot be produced in sufficient quantities as are now required for human health. Farmed fish have traditionally been fed a diet consisting of fishmeal and fish oil, rich in n-3 LC-PUFA. However, the increase in global aquaculture production has resulted in these finite and limited marine ingredients being replaced with sustainable alternatives of terrestrial origin that are devoid of n-3 LC-PUFA. Consequently, the nutritional value of the final product has been partially compromised with EPA and DHA levels both falling. Recent calls from the salmon industry for new sources of n-3 LC-PUFA have received significant commercial interest. Thus, this review explores the technologies being applied to produce de novo n-3 LC-PUFA sources, namely microalgae and genetically engineered oilseed crops, and how they may be used in aquafeeds to ensure that farmed fish remain a healthy component of the human diet.     

Effect of dietary lipids on plasma fatty acid profiles and prostaglandin and leptin production in gilthead seabream (Sparus aurata)

The aim of this study was to investigate the effects of different levels of substitution of fish oil by vegetable oils rich in oleic, linoleic and linolenic acids on gilthead seabream plasma and leukocyte fatty acid compositions and prostaglandin (PG) and leptin production. Juvenile seabream of 24 g initial body mass were fed four iso-energetic and iso-proteic experimental diets for 281 days. Fatty acid composition of plasma lipids was markedly affected by the inclusion of vegetable oils (VO). ARA (arachidonate), EPA (eicosapentaenoate) and DHA (docosahexaenoate) were preferentially incorporated into polar lipids of plasma, and DHGLA (di-homogammalinoleate) accumulated with increased vegetable oil inclusion. Dietary treatments resulted in alterations of DHGLA/ARA ratios, but not ARA/EPA. ARA-derived PGE₂ production in plasma was not affected by vegetable oils, in agreement with similar eicosanoid precursor ratio (ARA/EPA) in leukocytes total lipids and plasma phospholipids among fish fed with the different dietary treatments. Feeding vegetable oils leads to a decrease in plasma EPA which in turn reduced plasma PGE₃ concentration. Moreover, PGE₃ was the major prostaglandin produced in plasma of fish fed fish oil based diet. Such findings point out the importance of EPA as a precursor of prostaglandins in marine fish, at least for the correct function of the blood cells, and correlates well with the predominant role of this fatty acid in immune regulation in this species. A negative correlation was found between plasma PGE₂ and leptin plasma concentration, suggesting that circulating levels of leptin may act as a metabolic signal modulating PGE₂ release. The present study has shown that increased inclusion of vegetable oils in diet for gilthead seabream may profoundly affect the fatty acid composition of plasma and leukocytes, specially HUFA (highly unsaturated fatty acids), and consequently the production of PGE₃, which can be a major PG in plasma. Alteration in the amount and type of PG produced can be at least partially responsible for the changes in the immune system and health parameters of fish fed diets with high inclusion of VO.     

Engineering oilseeds to produce nutritional fatty acids

There is a growing body of evidence suggesting that regular consumption of foods rich in omega-3 long chain polyunsaturated fatty acids has multiple positive health benefits. The fats and oils from marine fish contain high contents of these beneficial fatty acids but increased consumer demand has also increased strain on the ability of the world's fisheries to meet demand from wild capture. Many consumers are choosing fish oil supplements or are eating foods that have been complemented with fish oils instead of consuming fish directly. However, removing undesirable odors, flavors and contaminants is expensive. In contrast, oils derived from land plants such as soybean are inexpensive and contaminant free. Recent strides in plant molecular biology now allow the engineering of oilseeds for the production of novel fats and oils, including those synthesized by complex, multigene biosynthetic pathways such as the omega-3 long chain polyunsaturated fatty acids. Given the potential benefits to the environment with regards to overfishing and the health prospects of increased consumption of these healthy fatty acids, producing these fatty acids in oilseeds is a desirable and worthy goal. In this review, we will describe the recent advances in this field along with some of the technical hurdles encountered thus far.     

Mechanism and challenges in commercialisation of algal biofuels

Biofuels made from algal biomass are being considered as the most suitable alternative energy in current global and economical scenario. Microalgae are known to produce and accumulate lipids within their cell mass which is similar to those found in many vegetable oils. The efficient lipid producer algae cell mass has been reported to contain more than 30% of their cell weight as lipids. According to US DOE microalgae have the potential to produce 100 times more oil per acre land than any terrestrial plants. This article reviews up to date literature on the composition of algae, mechanism of oil droplets, triacylglycerol (TAG) production in algal biomass, research and development made in the cultivation of algal biomass, harvesting strategies, and recovery of lipids from algal mass. The economical challenges in the production of biofuels from algal biomass have been discussed in view of the future prospects in the commercialisation of algal fuels.     

Laurie oil resources

Lauric oils and their derivatives have many applications both in the food and chemical industries. The major sources and some alternative raw materials for this multi-billon dollar business are discussed in the light of their ability to supply future market needs. There should be ample supply of lauric oils—except when drought and possibly disease affect large areas of coconut plantations—because of the rapid increase in palm kernel oil production expected from the African oil palm during the next decade. Most other sources are unlikely to be important in the short term because of the generally adequate supply of lauric oils and the considerable amount of research still needed to convert the best options into viable crops. However, a dramatic effect on supply can be expected if it becomes possible to manipulate the appropriate genes from lauric oil producing species of Cuphea into a conventional oil crop like rape. Future demand for lauric oils will be affected by the relative price of other vegetable oils and petroleum feedstocks that can be used to replace them in the manufacture of an increasing number of end products in both the food and chemical industries.     

An alternative to fish oils: Metabolic engineering of oil-seed crops to produce omega-3 long chain polyunsaturated fatty acids

It is now accepted that omega-3 polyunsaturated fatty acids, especially eicosapentaenoic acid (EPA; 20:5Δ5,8,11,14,17) and docosahexaenoic acid (DHA, 22:6Δ4,7,10,13,16,19) play important roles in a number of aspects of human health, with marine fish rich in these beneficial fatty acids our primary dietary source. However, over-fishing and concerns about pollution of the marine environment indicate a need to develop alternative, sustainable sources of very long chain polyunsaturated fatty acids (VLC-PUFAs) such as EPA and DHA. A number of different strategies have been considered, with one of the most promising being transgenic plants “reverse-engineered” to produce these so-called fish oils. Considerable progress has been made towards this goal and in this review we will outline the recent achievements in demonstrating the production of omega-3 VLC-PUFAs in transgenic plants. We will also consider how these enriched oils will allow the development of nutritionally-enhanced food products, suitable either for direct human ingestion or for use as an animal feedstuff. In particular, the requirements of aquaculture for omega-3 VLC-PUFAs will act as a strong driver for the development of such products. In addition, biotechnological research on the synthesis of VLC-PUFAs has provided new insights into the complexities of acyl-channelling and triacylglycerol biosynthesis in higher plants.     

Influence of extraction solvent system on the extractability of lipid components from the biomass of Nannochloropsis gaditana

Microalgae oils are considered to be promising alternative sources of omega-3 LC-PUFA. The aim of this work was therefore to evaluate different solvent (mixtures), currently accepted for use in the food industry, for the extraction of lipids from Nannochloropsis gaditana, an omega-3 LC-PUFA-rich microalga. Importantly, not only the total lipid yield but also the lipid class, eicosapentaenoic acid, carotenoid, and sterol yield were investigated. It was shown that the highest yield for each of the components was obtained with dichloromethane/ethanol (1:1). All extracts except the one obtained with dichloromethane/ethanol (1:1) were enriched in neutral lipids and depleted in polar lipids, when compared to the total lipid extract (chloroform/methanol 1:1). Hexane/isopropanol (3:2) seems to be the second best option: it has the advantage of performing better at criteria such as toxicity, but has the disadvantage that almost half of the interesting oil cannot be recovered.     

Stimulation of anaphylaxis in the mouse footpad by dietary fish oil fatty acids

The effect of fish oil-derived omega-3 (omega-3) fatty acids on anaphylaxis, Arthus and delayed type hypersensitivity reactions in mice has been investigated. Mice on a normal chow diet were fed eicosapentaenoic acid and docosahexaenoic acid at a dose of 500 and 333 mg/kg/day, respectively, by a gastric tube over a period of 61 days. Control groups were given water, safflower oil or oleic acid. Anaphylactic and Arthus type reactions were induced in the mouse footpad using bovine serum albumin as an antigen. Carrageenin was utilized to produce a delayed type hypersensitivity reaction. The animals fed omega-3 fatty acids induced a more anaphylactic foodpad reaction. There was no significant effect of the diet on Arthus and delayed type hypersensitivity responses. There was no effect of the fish oil-supplemented diet on production of antibodies to bovine serum albumin. Synthesis of prostaglandin E2 by peritoneal macrophages was significantly inhibited in the animals fed omega-3 fatty acid-enriched fish oil, while leukotriene B4 production was not affected. These results suggest that a diet enriched in omega-3 fatty acids modulates production of arachidonic acid metabolites and this may influence anaphylaxis, but not Arthus and cellular mediated hypersensitivity responses.     

Integrated Environmental Assessment of sunflower oil production

The high energy and petroleum based fuels demand of the world require use of alternative fuels from materials available within each country. Therefore biofuels may be an alternative to mineral diesel. A local pilot production of sunflower oil was implemented in order to test the possibility to reduce fossil fuels consumption on a local scale. Vegetable oils can be directly obtained from oilseed plants and can be used as energy sources in internal combustion engines.Environmental Impact Indicators were provided by Material Flow Accounting, Embodied Energy Analysis and Emergy Accounting. All three approaches reveal that the agricultural phase is the critical step in the whole production line from the point of view of environmental impact. The renewability calculated for the sunflower oil is 33.6%.A comparative Life Cycle Assessment analysis for the sunflower oil production line from conventional farming with organic farming showed environmental advantages.Furthermore use of vegetable oils instead of diesel oil as energy sources in internal combustion engines reduces carbon dioxide emissions of 59%.This study represents an opportunity for farms to reduce dependence on petroleum and to explore energy production systems exploiting renewable energy.     

Enzyme catalysed production of vegetable oils containing omega-3 polyunsaturated fatty acid

Summary The application of enzymatic interesterification for production of vegetable oils containing omega-3 polyunsaturated fatty acids was investigated. Six veteable oils were used as substrates, together with omega-3 polyunsaturated fatty acid, and reactions were catalysed by immobilized Mucor miehei lipase in organic solvent. The degree of incorporation of eicosapentaenoic acid and docosahexaenoic acid into corn oil, sunflower oil, peanut oil, olive oil and soybean oil were 17.71, 17.59, 16.79, 14.89, 13.91 and 10.48%, respectively, after a 12 h incubation period.     

Dietary fish oils modify adipocyte structure and function.

Dietary fish oils, enriched with omega-3 fatty acids (e.g., MaxEPA fish oil), inhibit lipogenesis and have a marked hypotriglyceridemic effect in man and experimental animals. Dietary omega-3 fatty acids also reduce adipose tissue trophic growth in rats. To understand the metabolic basis for this, we measured the effect of fish oil feeding upon rat plasma triglyceride concentration, fat pad mass, fat cell size, fat cell lipolysis, as well as lipoprotein binding to adipocyte plasma membranes. In adolescent (250 g) male Wistar rats fed 20% (w/w) fish oil supplemented diets for 3 weeks, plasma triglyceride levels and epididymal and perirenal fat pad mass were significantly (P less than 0.005) reduced compared to pair-fed controls given 20% lard diets. These differences in fat pad mass between the diets were greater than differences in whole animal mass or in the mass of livers, testes, kidneys, spleens, or hearts. Isoproterenol-stimulated lipolysis was significantly (P less than 0.005) higher in fish oil fed rats than in pair-fed controls. In young (100 g) rats plasma triglyceride levels were 10 times lower in the fish oil fed group after 5 weeks as compared to the lard-fed controls. This was accompanied by a reduction in epididymal and perirenal fat pad mass as well as a 2-3-fold decrease in adipocyte volumes; there was no significant difference between the two groups in fat cell number in each region. Plasma membranes of epididymal adipocytes from fish oil fed rats bound significantly (P less than 0.001) less HDL1 than the lard-fed rats, possibly as a result of a reduction in fat cell size and/or alteration of plasma membrane structure. Thus in both young and old rats, the reduction in plasma triglyceride concentration in conjunction with increased hormone-stimulated lipolysis may explain in part the selective reduction in adipose tissue trophic growth accompanying fish oil consumption.     

Fatty acid alkyl esters: perspectives for production of alternative biofuels

The global economy heads for a severe energy crisis: whereas the energy demand is going to rise, easily accessible sources of crude oil are expected to be depleted in only 10–20 years. Since a serious decline of oil supply and an associated collapse of the economy might be reality very soon, alternative energies and also biofuels that replace fossil fuels must be established. In addition, these alternatives should not further impair the environment and climate. About 90% of the biofuel market is currently captured by bioethanol and biodiesel. Biodiesel is composed of fatty acid alkyl esters (FAAE) and can be synthesized by chemical, enzymatic, or in vivo catalysis mainly from renewable resources. Biodiesel is already established as it is compatible with the existing fuel infrastructure, non-toxic, and has superior combustion characteristics than fossil diesel; and in 2008, the global production was 12.2 million tons. The biotechnological production of FAAE from low cost and abundant feedstocks like biomass will enable an appreciable substitution of petroleum diesel. To overcome high costs for immobilized enzymes, the in vivo synthesis of FAAE using bacteria represents a promising approach. This article points to the potential of different FAAE as alternative biofuels, e.g., by comparing their fuel properties. In addition to conventional production processes, this review presents natural and genetically engineered biological systems capable of in vivo FAAE synthesis.