Comparative life cycle assessment of a commercial algal multiproduct biorefinery and wild caught fishery for small pelagic fish

Purpose

The purpose of this study was to compare the environmental impacts of omega-3 fatty acid (n-3), high protein feed and biofuel production from algae to the impacts of the production of those products from fish.

Methods

The functional unit was the production of one metric ton of omega-3 fatty acids from algae (fish) and the accompanying co-products of biofuel and high protein feed. This was a cradle to gate LCA. Four scenarios were used in this model. The algae multiproduct model (MPM) scenario was the baseline using only unit operations currently in use at the reference facility (Cellana LLC). A low-energy centrifuge replaced the existing conventional centrifuge (MPM (LE)) to reduce energy consumption. The MPM was improved in a different manner, employing membrane filtration prior to centrifugation (MPM (MF)). These three scenarios were compared to the conventional production of the same products from fish (conventional product model: CPM). This life cycle assessment investigated the following impacts: ozone depletion potential, global warming potential, smog formation potential, acidification potential, and eutrophication potential.

Results and discussion

The environmental impacts of producing omega-3 fatty acids from algae were higher than producing omega-3 fatty acids from fish if membrane filtration was not used. Membrane filtration reduced most of the environmental impacts of the algae system by more than 50%. Fuel consumption was the only factor that caused the fish systems to change by greater than 10% from the baseline. Productivity, membrane filtration electricity, and annual operating days could each affect the environmental impacts of the algae system by greater than 10% from the baseline. Improvements to the algae system depend on improvements to cultivation and harvesting, with the impacts from processing being very small.

Conclusions

This study presented results comparing the environmental impacts from a multiproduct system from algae and from fish. The results of this study can serve as a benchmark for the environmental impacts of an algal multiproduct biorefinery compared to the conventional production of those same products from fish. Areas of improvement have been identified for the algae production system for dewatering and cultivation. The amount of n-3 had little impact on the n-3 market but had a significant effect on the existing algal n-3 market. The amount of fuel and feed produced had a negligible effect on both markets.

     

Fatty acids filamentous green algae

Fatty acid analyses of several filamentous green algae were conducted using gas-liquid chromatography. Two bryophytes were also examined. Qualitatively, the genera of algae studied were divided into two groups: (A) algae that have significant amounts of polyunsaturated C₂₀ fatty acids and (B) algae that lack or only have very small amounts of the C₂₀ acids. On the basis of fatty acid content, the algae of Group A more closely resemble the bryophytes than do the algae of Group B. Culture age was shown to cause quantitative but not qualitative variations in fatty acid content. It is evident from this study that extrapolation to land plants, from studies on the fatty acid content of the green algae, should include the filamentous forms.     

Snow and Glacial Algae: A Review1

Snow or glacial algae are found on all continents, and most species are in the Chlamydomonadales (Chlorophyta) and Zygnematales (Streptophyta). Other algal groups include euglenoids, cryptomonads, chrysophytes, dinoflagellates, and cyanobacteria. They may live under extreme conditions of temperatures near 0°C, high irradiance levels in open exposures, low irradiance levels under tree canopies or deep in snow, acidic pH, low conductivity, and desiccation after snow melt. These primary producers may color snow green, golden-brown, red, pink, orange, or purple-grey, and they are part of communities that include other eukaryotes, bacteria, archaea, viruses, and fungi. They are an important component of the global biosphere and carbon and water cycles. Life cycles in the Chlamydomonas–Chloromonas–Chlainomonas complex include migration of flagellates in liquid water and formation of resistant cysts, many of which were identified previously as other algae. Species differentiation has been updated through the use of metagenomics, lipidomics, high-throughput sequencing (HTS), multi-gene analysis, and ITS. Secondary metabolites (astaxanthin in snow algae and purpurogallin in glacial algae) protect chloroplasts and nuclei from damaging PAR and UV, and ice binding proteins (IBPs) and polyunsaturated fatty acids (PUFAs) reduce cell damage in subfreezing temperatures. Molecular phylogenies reveal that snow algae in the Chlamydomonas–Chloromonas complex have invaded the snow habitat at least twice, and some species are polyphyletic. Snow and glacial algae reduce albedo, accelerate the melt of snowpacks and glaciers, and are used to monitor climate change. Selected strains of these algae have potential for producing food or fuel products.     

Extraction of pigments and fatty acids from the green alga Scenedesmus obliquus (Chlorophyceae)

In this paper, the efficiency of pigment and fatty acid extraction from resistant algae using Scenedesmus obliquus as an example was examined. We found that adding quartz sand and solvent to freeze-dried algal material and subsequent extraction in an ultrasound bath for 90min at –4°C resulted in excellent extraction of these compounds. This extraction method was compared with a method regularly used for extraction of fatty acids and pigments, i.e. addition of solvents to algal material with subsequent incubation. Our extraction using the ultrasound and sand method was about twice as efficient as this method for both pigments and fatty acids. The ultrasound method is simple, extracts over 90% of the different substances in one step and conserves the relationships of pigments and fatty acids. In addition, no alteration- or breakdown products were observed with the new method. Thus, this method allows accurate quantitative extraction of both pigments and fatty acids from Scenedesmus obliquus and other algae. The method was also been found to be as effective for Cryptomonas erosa (Cryptophyceae), Cyclotella meneghiniana (Bacillariophyceae), Microcystis aeruginosa (Cyanophyceae), and Staurastrum paradoxum (Chlorophyceae, Desmidiaceae) and is thus applicable to a wide spectrum of algae.     

Lipid composition ofAcetabularia mediterranea (Lamour.) Variations during cap morphogenesis

Summary YoungAcetabularia mediterranea cells without cap have a fatty acid composition different from other green algae currently used for biological research. They contain important quantities of palmitic and oleic acid, but are very poor in polyunsaturated fatty acids such as linoleic and linolenic acid. (These polyunsaturated fatty acids are predominant in higher plants and many green algae.)     

Genomic and biochemical analysis of lipid biosynthesis in the unicellular rhodophyte Cyanidioschyzon merolae: lack of a plastidic desaturation pathway results in the coupled pathway of galactolipid synthesis

The acyl lipids making up the plastid membranes in plants and algae are highly enriched in polyunsaturated fatty acids and are synthesized by two distinct pathways, known as the prokaryotic and eukaryotic pathways, which are located within the plastids and the endoplasmic reticulum, respectively. Here we report the results of biochemical as well as genomic analyses of lipids and fatty acids in the unicellular rhodophyte Cyanidioschyzon merolae. All of the glycerolipids usually found in photosynthetic algae were found, such as mono- and digalactosyl diacylglycerol, sulfolipid, phosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol. However, the fatty acid composition was extremely simple. Only palmitic, stearic, oleic, and linoleic acids were found as major acids. In addition, 3-trans-hexadecanoic acid was found as a very minor component in phosphatidylglycerol. Unlike the case for most other photosynthetic eukaryotes, polyenoic fatty acids having three or more double bonds were not detected. These results suggest that polyunsaturated fatty acids are not necessary for photosynthesis in eukaryotes. Genomic analysis suggested that C. merolae lacks acyl lipid desaturases of cyanobacterial origin as well as stearoyl acyl carrier protein desaturase, both of which are major desaturases in plants and green algae. The results of labeling experiments with radioactive acetate showed that the desaturation leading to linoleic acid synthesis occurs on phosphatidylcholine located outside the plastids. Monogalactosyl diacylglycerol is therefore synthesized by the coupled pathway, using plastid-derived palmitic acid and endoplasmic reticulum-derived linoleic acid. These results highlight essential differences in lipid biosynthetic pathways between the red algae and the green lineage, which includes plants and green algae.     

A review of nutritional effects on fat composition of animal products with special emphasis on n-3 polyunsaturated fatty acids

The fatty acid composition of animal products (eggs, milk and meat) is the reflect of both the tissue fatty acid biosynthesis and the fatty acid composition of ingested lipids. This relationship is stronger in monogastrics (pigs, poultry and rabbits) than in ruminants, where dietary fatty acids are hydrogenated in the rumen. There is an increasing recognition of the health benefits of polyunsaturated fatty acids (PUFA), because these fatty acids are essential for humans. In addition, the ratio n-6/n-3 fatty acids in the human diet is important. This ratio by far exceeds the recommended value of 5. Therefore, inclusion of fish meals, or n-3 PUFA rich oils, or linseed in animal diets is a valid means of meeting consumer demand for animal products that are nutritionally beneficial.The studies that are undertaken on animals mainly use diets supplemented with linseed, as a source of n-3 fatty acids. The use of linseed diets generally leads to an increased n-3 fatty acid content in animal products (egg, meat, milk) in ruminants and monogastrics. Recent studies have also demonstrated that neither the processing nor the cooking affects the PUFA content of pork meat or meat products.The ability of unsaturated fatty acids, especially those with more than two double bonds, to rapidly oxidise, is important in regulating the shelf life of animal products (rancidity and colour deterioration); however, a good way to avoid such problems is to use antioxidant products (such as vitamin E) in the diet.Some studies also show that it is not necessary to feed animals with linseed-supplemented diets for a long time to have the highest increase in PUFA content of the products. So, short-term diet manipulation can be a practical reality for industry.As the market for n-3 PUFA enriched products is today limited in most countries, other studies must be undertaken to develop this kind of production.     

Mouse models for disorders of mitochondrial fatty acid beta-oxidation

Mitochondrial beta-oxidation of fatty acids is vital for energy production in periods of fasting and other metabolic stress. Human patients have been identified with inherited disorders of mitochondrial beta-oxidation of fatty acids with enzyme deficiencies identified at many of the steps in this pathway. Although these patients exhibit a range of disease processes, Reye-like illness (hypoketotic-hypoglycemia, hyperammonemia and fatty liver) and cardiomyopathy are common findings. There have been several mouse models developed to aid in the study of these disease conditions. The characterized mouse models include inherited deficiencies of very long-chain acyl-CoA dehydrogenase, long-chain acyl-CoA dehydrogenase, short-chain acyl-CoA dehydrogenase, mitochondrial trifunctional protein-alpha, and medium-/short-chain hydroxyacyl-CoA dehydrogenase. Mouse mutants developed, but presently incompletely characterized as models, include carnitine palmitoyltransferase-1a and medium-chain acyl-CoA dehydrogenase deficiencies. In general, the mouse models of disorders of mitochondrial fatty acid beta-oxidation have shown clinical signs that include Reye-like syndrome and cardiomyopathy, and many are cold intolerant. It is expected that these mouse models will provide vital contributions in understanding the mechanisms of disease pathogenesis of fatty acid oxidation disorders and the development of appropriate treatments and supportive care.     

Current progress in the fatty acid metabolism in Cryptosporidium parvum

Cryptosporidium parvum is one of the apicomplexans that can cause severe diarrhea in humans and animals. The slow development of anti-cryptosporidiosis chemotherapy is primarily due to the poor understanding on the basic metabolic pathways in this parasite. Many well-defined or promising drug targets found in other apicomplexans are either absent or highly divergent in C. parvum. The recently discovered apicoplast and its associated Type II fatty acid synthetic enzymes in Plasmodium, Toxoplasma, and Eimeria apicomplexans are absent in C. parvum, suggesting this parasite is unable to synthesize fatty acids de novo. However, C. parvum possesses a giant Type I fatty acid synthase (CpFAS1) that makes very long chain fatty acids using mediate or long chain fatty acids as precursors. Cryptosporidium also contains a Type I polyketide synthase (CpPKS1) that is probably involved in the production of unknown polyketide(s) from a fatty acid precursor. In addition to CpFAS1 and CpPKS1, a number of other enzymes involved in fatty acid metabolism have also been identified. These include a long chain fatty acyl elongase (LCE), a cytosolic acetyl-CoA carboxylase (ACCase), three acyl-CoA synthases (ACS), and an unusual "long-type" acyl-CoA binding protein (ACBP), which allows us to hypothetically reconstruct the highly streamlined fatty acid metabolism in this parasite. However, C. parvum lacks enzymes for the oxidation of fatty acids, indicating that fatty acids are not an energy source for this parasite. Since fatty acids are essential components of all biomembranes, molecular and functional studies on these critical enzymes would not only deepen our understanding on the basic metabolism in the parasites, but also point new directions for the drug discovery against C. parvum and other apicomplexan-based diseases.     

Natural halogenated fatty acids: their analogues and derivatives

A comprehensive survey has been made of all fatty acids containing halogen atoms covalently bonded to carbon and which are deemed as naturally occurring. Generally thought to be minor components produced by many different organisms, these interesting compounds now number more than 300. Recent research, especially in the marine area, indicates this number will increase in the future. Sources of halogenated fatty acids include microorganisms, algae, marine invertebrates, and higher plants and some animals. Their possible biological significance has also been discussed     

Photoheterotrophy in the production of phytoplankton organisms

Interest is growing in algae as sources of medicinal and other potentially useful compounds, as well as their use in fish rearing. We are interested in their production of polyunsaturated fatty acids (PUFA). Photoautotrophic growth gives the highest levels of unsaturation in the fatty acid pool, but biomass concentrations are low. Heterotrophy on sugars gives higher biomass but seems to give more saturation in the fatty acids. In freshwater algae acetate has proved to be a good carbon source for photoheterotrophic growth, giving a crop with reasonably high levels of PUFA. In addition it is possible to regulate acetic acid addition through the pH change as acetate is used up in a well-aerated system, so achieving high biomass yields in the presence of relatively low acetate concentration. When we attempted to extend this to marine algae (principally species used in fish farming), we found that acetic acid was ineffective or sometimes toxic to most species tested, even at high pH. However, glycerol stimulated growth in a number of the algae. We report on this stimulation, and on the fatty acid composition of the resulting algal crop, discuss the problems in regulating the addition of this metabolite to algal cultures, and speculate on applications in the production of other useful algal metabolites. We also show that some of these algae used in fish farming grow best when the salinity of the water is rather less than that found in standard sea water.     

Metabolic acidosis: separation methods and biological relevance of organic acids and lactic acid enantiomers

Metabolic acidosis can result from accumulation of organic acids in the blood due to anaerobic metabolism or intestinal bacterial fermentation of undigested substrate under certain conditions. These conditions include short-bowel syndrome, grain overfeeding of ruminants and, as recently reported, severe gastroenteritis. Measuring fermentation products such as short-chain fatty acids (SCFAs) and lactic acid in various biological samples is integral to the diagnosis of bacterial overgrowth. Stereospecific measurement of D- and L-lactic acid is necessary for confirmation of the origin and nature of metabolic acidosis. In this paper, methods for the separation of SCFAs and lactic acid are reviewed. Analysis of the organic acids involved in carbohydrate metabolism has been achieved by enzymatic methods, gas chromatography, high-performance liquid chromatography and capillary electrophoresis. Sample preparation techniques developed for these analytes are also discussed.     

Free and bound fatty acid oxidation products in archaeological ceramic vessels

While oxidation products of unsaturated fatty acids, for example dicarboxylic acids (hereafter diacids), must form during the use of unglazed ceramic vessels for the processing of animal and plant products, such components have never been observed during studies of absorbed lipids. Their absence from the extractable lipid fraction is presumed to be the result of their loss from potsherds through groundwater leaching. Lipid oxidation products including short-chain dicarboxylic acids, ω-hydroxy acids and longer-chain hydroxy and dihydroxy acids have now been observed as components probably covalently bound into solvent insoluble residues of potsherds recovered from waterlogged deposits. These components were only revealed following alkaline treatment of the insoluble residues. A similar mixture of diacids was observed in high abundance in the free lipid fraction of vessels recovered from an exceptionally arid deposit where groundwater leaching would never have occurred. These results confirm the formation of oxidation and probable polymerization products of unsaturated fatty acids during vessel use and burial.     

Hydroxy long-chain fatty acids in fungi

Hydroxy long-chain fatty acids occur widely in animals and plants and have important physiological activities in these eukaryotes. There are indications that these compounds are also common and important in fungi. The occurrence of hydroxy-polyunsaturated fatty acids (hydroxy-PUFAs) is of biotechnological importance, because these compounds are potentially high-value lipid products with medical applications. This review pays particular attention to the production of hydroxy-PUFAs by yeasts and other fungi. Hydroxy-PUFAs derived from lipoxygenase activity appear to be present in most fungi, while hydroxy-PUFAs from cyclooxygenase activity (i.e. prostaglandins) have mainly been implicated in the Oomycota and in yeasts from the genus Dipodascopsis. The occurrence of other hydroxy long-chain fatty acids in fungi is also discussed briefly; these include hydroxy fatty acids that are generally associated with cytochrome P-450 monooxygenase activity (i.e. terminal and sub-terminal hydroxy acids and diols derived from the corresponding epoxides) as well as 2-hydroxy-fatty acids and 3-hydroxy-fatty acids.The authors are with the Department of Microbiology and Biochemistry, University of the Orange Free State, P.O. Box 339, Bloemfontein, 9300, South Africa     

Alteration of the binding specificity of cellular retinol-binding protein II by site-directed mutagenesis.

Rat cellular retinol-binding protein II (CRBP II) is an abundant 134-residue intestinal protein that binds all-trans-retinol and all-trans-retinal. It belongs to a family of homologous, 15-kDa cytoplasmic proteins that bind hydrophobic ligands in a noncovalent fashion. These binding proteins include a number of proteins that bind long chain fatty acids. X-ray analyses of the structure of two family members, rat intestinal fatty acid-binding protein and bovine myelin P2 protein, indicate that they have a high degree of conformational similarity and that the carboxylate group of their bound fatty acid interacts with a delta-guanidium group of at least 1 of 2 "buried" arginine residues. These 2 Arg residues are conserved in other family members that bind long chain fatty acids and in cellular retinoic acid-binding protein, but are replaced by Gln109 and Gln129 in CRBP II. We have genetically engineered two amino acid substitutions in CRBP II: 1) Gln109 to Arg and 2) Gln129 to Arg. The purified Escherichia coli-derived CRBP II mutant proteins were analyzed by fluorescence and nuclear magnetic resonance spectroscopy. Both mutants exhibit markedly decreased binding of all-trans-retinol and all-trans-retinaldehyde, but no increased binding of all-trans-retinoic acid. Arg substitution for Gln109 but not for Gln129 produces a dramatic increase in palmitate binding activity. Analysis of the endogenous fatty acids associated with the purified E. coli-derived proteins revealed that E. coli-derived intestinal fatty acid binding protein and the Arg109 CRBP II mutant are complexed with endogenous fatty acids in a qualitatively and quantitatively similar manner. These results provide evidence that this internal Arg may play an important role in the binding of long chain fatty acids by members of this protein family.     

Cell signal mechanisms,conjugated linoleic acids (CLAs) and anti-tumorigenesis

Fats have been adversely implicated in the aetiology of many forms of cancer yet evidence is accumulating that certain types of fatty acids have anticancer properties. This is well documented for fish-oil fatty acids of the n-3 family. Recently, fatty acids found to occur naturally in ruminant-derived food products were found to have anticancer properties. These fatty acids were identified as conjugated linoleic acids (CLAs) derived from the parent linoleic acid by its partial hydrogenation by rumen bacteria. Studies with tumour-bearing animals have shown that consumption of CLAs particularly with regard to breast and prostate cancer is beneficial. Studies with cancer cells have also shown that these fatty acids can inhibit cell proliferation and induce cell death. However, little is known regarding the mechanisms of action of these CLAs. In particular, which cellular signal mechanisms are regulated by CLAs which can explain their anticancer properties. We have shown that CLAs specifically up-regulate cell signal systems at the level of gene expression (mRNA, protein) in human breast and prostate cancer cells which are responsible for the induction of apoptosis or programmed cell death. These findings support the anticancer effects of CLA found in animal models and indicate similar effects could occur in man.     

Lipid and fatty acid composition of the green oleaginous alga Parietochloris incisa,the richest plant source of arachidonic acid

We have hypothesized that among algae of alpine environment there could be strains particularly rich in long chain polyunsaturated fatty acids (LC-PUFA). Indeed, the chlorophyte (Trebuxiophyceae) Parietochloris incisa isolated from Mt. Tateyama, Japan, was found to be the richest plant source of the pharmaceutically valuable LC-PUFA, arachidonic acid (AA, 20:4omega6). The alga is also extremely rich in triacylglycerols (TAG), which reaches 43% (of total fatty acids) in the logarithmic phase and up to 77% in the stationary phase. In contrast to most algae whose TAG are made of mainly saturated and monounsaturated fatty acids, TAG of P. incisa are the major lipid class where AA is deposited, reaching up to 47% in the stationary phase. Except for the presence of AA, the PUFA composition of the chloroplastic lipids resembled that of green algae, consisting predominantly of C(16) and C(18) PUFAs. The composition of the extrachloroplastic lipids is rare, including phosphatidylcholine (PC), phosphatidylethanolamine (PE) as well as diacylglyceryltrimethylhomoserine (DGTS). PC and PE are particularly rich in AA and are also the major depots of the presumed precursors of AA, l8:3omega6 and 20:3omega6, respectively.     

Algae as promising organisms for environment and health

Algae, like other plants, produce a variety of remarkable compounds collectively referred to as secondary metabolites. They are synthesized by these organisms at the end of the growth phase and/or due to metabolic alterations induced by environmental stress conditions. Carotenoids, phenolic compounds, phycobiliprotein pigments, polysaccharides and unsaturated fatty acids are same of the algal natural products, which were reported to have variable biological activities, including antioxidant activity, anticancer activity, antimicroabial activity against bacteria-virus-algae-fungi, organic fertilizer and bioremediation potentials.Key words: algae, biological activities, active ingredients