Reversed-Phase Medium-Pressure Liquid Chromatography Purification of Omega-3 Fatty Acid Ethyl Esters Using AQ-C18.

Omega-3 fatty acids are in high demand due to their efficacy in treating hypertriglyceridemia and preventing cardiovascular diseases. However, the growth of the industry is hampered by low purity and insufficient productivity. This study aims to develop an efficient RP-MPLC purification method for omega-3 fatty acid ethyl esters with high purity and capacity. The results indicate that the AQ-C18 featuring polar end-capped silanol groups outperformed C18 and others in retention time and impurity separation. By injecting pure fish oil esters with a volume equivalent to a 1.25% bed volume on an AQ-C18 MPLC column using a binary isocratic methanol-water (90:10, v:v) mobile phase at 30 mL/min, optimal omega-3 fatty acid ethyl esters were obtained, with the notable purity of 90.34% and a recovery rate of 74.30%. The total content of EPA and DHA produced increased from 67.91% to 85.27%, meeting the acceptance criteria of no less than 84% set by the 2020 edition of the Pharmacopoeia of the People's Republic of China. In contrast, RP-MPLC significantly enhanced the production efficiency per unit output compared to RP-HPLC. This study demonstrates a pioneering approach to producing omega-3 fatty acid ethyl esters with high purity and of greater quantity using AQ-C18 RP-MPLC, showing this method's significant potential for use in industrial-scale manufacturing.

Supercritical CO2 extraction increases the recovery levels of omega-3 fatty acids in Tetraselmis chuii extracts.

Tetraselmis chuii is a microalgae marketed as ingredient meeting the acceptance criteria for novel foods established by the European Union and can be an important source of healthy fatty acids (FA). The aim of this research was to characterize the FA profile of T. chuii fractions obtained by supercritical carbon dioxide (SCCO2) extraction operating with two sequential co-solvents and to evaluate the effect of biomass pretreatment (freeze/thaw cycles followed by ultrasounds). T. chuii biomass was confirmed to be an important source of omega-3 FA, mainly due to the abundance of α-linolenic acid, and pre-treatment significantly improved the lipid yield. Other omega-3 FA, such as 16:3, 16:4, 18:4, 18:5, 20:3 and 20:5, were also detected in different proportions. When SCCO2 extraction of pretreated and un-pretreated T. chuii was compared with conventional solvent extraction, the nutritional quality indices of the extracts were improved by the use of SCCO2.

Variability of Omega-3/6 Fatty Acid Obtained Through Extraction-Transesterification Processes from Phaeodactylum tricornutum.

The effect of direct transesterification methods on the omega-3/6 composition of extracts from Phaeodactylum tricornutum was studied. The aim of this work was to identify an extraction method which allowed to obtain the most suitable profile of fatty acids in terms of its potential benefits to health, particularly if further used in the food industry. Seven methods using acids, alkalis, and heterogeneous-catalysts, (namely methods from 1 to 7, abbreviated as M1-M7) were performed to determine α-linolenic (ALA), linoleic (LA), docosahexaenoic (DHA) and eicosapentaenoic (EPA) acids. The composition of fatty acids was in all cases characterized by the major abundance of palmitic (23.95-34.08%), palmitoleic (30.94-35.56%), oleic acids (3.00-7.41%), and EPA (0.5-6.45%). Unsaturated fatty acids extraction yield was higher with a two-step transesterification process (M6, 63.65%). The total fatty acid methyl ester content (FAME) obtained with acid-transesterification (M1) reached about 21% wt, and 60% w/w total lipids. ALA higher relative content (ALA/LA ratio) was obtained when a lipid pre-extraction step was performed prior to acid-catalysis (M4). The transesterification method based on alkali-catalyst (M3, KOH catalyst) led to obtain higher DHA relative contents (DHA/EPA ratio up to 0.11), although its FAME content was 3.75-fold lower than that obtained with acid-transesterification (M1). Overall, this study shows that direct transesterification with alkali-catalyst (M3) improves the determination of PUFA content from the diatom through a more efficient transesterification-based extraction process, and thus allow to assess the value of the biomass more accurately for application in the food industry.

A metabolomics comparison of plant-based meat and grass-fed meat indicates large nutritional differences despite comparable Nutrition Facts panels.

A new generation of plant-based meat alternatives-formulated to mimic the taste and nutritional composition of red meat-have attracted considerable consumer interest, research attention, and media coverage. This has raised questions of whether plant-based meat alternatives represent proper nutritional replacements to animal meat. The goal of our study was to use untargeted metabolomics to provide an in-depth comparison of the metabolite profiles a popular plant-based meat alternative (n = 18) and grass-fed ground beef (n = 18) matched for serving size (113 g) and fat content (14 g). Despite apparent similarities based on Nutrition Facts panels, our metabolomics analysis found that metabolite abundances between the plant-based meat alternative and grass-fed ground beef differed by 90% (171 out of 190 profiled metabolites; false discovery rate adjusted p < 0.05). Several metabolites were found either exclusively (22 metabolites) or in greater quantities in beef (51 metabolites) (all, p < 0.05). Nutrients such as docosahexaenoic acid (ω-3), niacinamide (vitamin B3), glucosamine, hydroxyproline and the anti-oxidants allantoin, anserine, cysteamine, spermine, and squalene were amongst those only found in beef. Several other metabolites were found exclusively (31 metabolites) or in greater quantities (67 metabolites) in the plant-based meat alternative (all, p < 0.05). Ascorbate (vitamin C), phytosterols, and several phenolic anti-oxidants such as loganin, sulfurol, syringic acid, tyrosol, and vanillic acid were amongst those only found in the plant-based meat alternative. Large differences in metabolites within various nutrient classes (e.g., amino acids, dipeptides, vitamins, phenols, tocopherols, and fatty acids) with physiological, anti-inflammatory, and/or immunomodulatory roles indicate that these products should not be viewed as truly nutritionally interchangeable, but could be viewed as complementary in terms of provided nutrients. The new information we provide is important for making informed decisions by consumers and health professionals. It cannot be determined from our data if either source is healthier to consume.

In vitro intestinal digestion of lipids from the marine diatom Porosira glacialis compared to commercial LC n-3 PUFA products.

Marine sources of long chain omega-3 polyunsaturated fatty acids (LC n-3 PUFA) are in high demand for use in health supplements. Mass cultivated marine microalgae is a promising and sustainable source of LC n-3 PUFA, which relieves pressure on natural fish stocks. The lipid class profile from cultivated photosynthetic algae differ from the marine organisms currently used for the production of LC n-3 PUFA. The objective of this study was to compare in vitro intestinal digestion of oil extracted from the cold-adapted marine diatom Porosira glacialis with commercially available LC n-3 PUFA supplements; cod liver oil, krill oil, ethyl ester concentrate, and oil from the copepod Calanus finmarchicus (Calanus® oil). The changes in the free fatty acids and neutral and polar lipids during the enzymatic hydrolysis were characterized by liquid and gas chromatography. In Calanus® oil and the Ethyl ester concentrate, the free fatty acids increased very little (4.0 and 4.6%, respectively) during digestion. In comparison, free fatty acids in Krill oil and P. glacialis oil increased by 14.7 and 17.0%, respectively. Cod liver oil had the highest increase (28.2%) in free fatty acids during the digestion. Monounsaturated and saturated fatty acids were more easily released than polyunsaturated fatty acids in all five oils.

Shrimp Oil Extracted from Shrimp Processing By-Product Is a Rich Source of Omega-3 Fatty Acids and Astaxanthin-Esters, and Reveals Potential Anti-Adipogenic Effects in 3T3-L1 Adipocytes.

The province of Newfoundland and Labrador, Canada, generates tons of shrimp processing by-product every year. Shrimp contains omega (n)-3 polyunsaturated fatty acids (PUFA) and astaxanthin (Astx), a potent antioxidant that exists in either free or esterified form (Astx-E). In this study, shrimp oil (SO) was extracted from the shrimp processing by-product using the Soxhlet method (hexane:acetone 2:3). The extracted SO was rich in phospholipids, n-3 PUFA, and Astx-E. The 3T3-L1 preadipocytes were differentiated to mature adipocytes in the presence or absence of various treatments for 8 days. The effects of SO were then investigated on fat accumulation, and the mRNA expression of genes involved in adipogenesis and lipogenesis in 3T3-L1 cells. The effects of fish oil (FO), in combination with Astx-E, on fat accumulation, and the mRNA expression of genes involved in adipogenesis and lipogenesis were also investigated. The SO decreased fat accumulation, compared to untreated cells, which coincided with lower mRNA expression of adipogenic and lipogenic genes. However, FO and FO + Astx-E increased fat accumulation, along with increased mRNA expression of adipogenic and lipogenic genes, and glucose transporter type 4 (Glut-4), compared to untreated cells. These findings have demonstrated that the SO is a rich source of n-3 PUFA and Astx-E, and has the potential to elicit anti-adipogenic effects. Moreover, the SO and FO appear to regulate adipogenesis and lipogenesis via independent pathways in 3T3-L1 cells.

Farmed Gilthead Sea Bream (Sparus aurata) by-Products Valorization: Viscera Oil ω-3 Enrichment by Short-Path Distillation and In Vitro Bioactivity Evaluation.

This study shows a pilot scale protocol aimed to obtain an omega 3-enriched oil after the processing of farmed gilthead sea bream viscera (SBV); this was oil was tested in vitro for bioactivity, attesting to the possibility to turn waste into profit The quality of the oil, in terms of requirements for animal and human consumption, was assessed by determining some chemical parameters, such as peroxide value (PV), thiobarbituric acid reactive substances (TBARS), ρ-anisidine (ρ-AV) content, total oxidation value (TOTOX), and phospholipids and free fatty acid (%), both in crude viscera oil (CVO) and refined viscera oil (RVO). Among the extraction conditions, the higher CVO yields were obtained at 60 °C for 10 min (57.89%) and at 80 °C for 10 min (67.5%), and the resulting oxidation levels were low when utilizing both extraction conditions. RVO, obtained from CVO extracted at 60 °C, showed the highest quality on the basis of the assessed parameters. The ethyl esters of the total fatty acid (TFA) contents extracted from RVO were enriched in the ω-3 polyunsaturated fatty acid fraction (PUFAE) up to almost 56% via short path distillation (SPD). Antioxidant activities and adipogenic properties were tested in vitro. PUFAE protected 3T3 L1 cells from oxidative stress and exerted an anti-adipogenic effect in Dicentrarchus labrax pre-adipocytes, attesting to the beneficial properties for both farmed fish and human health. These results could stimulate the adoption of solutions aimed to recover and utilize aquaculture by-products at a higher scale, turning "waste into profit" and indicating a strategy to reach more sustainable business models in aquaculture resource utilization according to the principles of the circular economy.

Simultaneous Treatment of Long-chain Monounsaturated Fatty Acid and n-3 Polyunsaturated Fatty Acid Decreases Lipid and Cholesterol Levels in HepG2 Cell.

The n-3 type polyunsaturated fatty acids (n-3PUFAs), including eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), from fish oil exhibit health benefits such as triacylglycerol- and cholesterol-lowering effects. Some pelagic fishes contain long-chain monounsaturated fatty acids (LC-MUFAs) such as eicosenoic acid (C20:1), which exert health-promoting effects. However, no study has evaluated beneficial effects of n-3PUFA and LC-MUFA combination. Here, we investigated effects of simultaneous treatment with n-3PUFA (EPA and DHA) and LC-MUFA (cis-5-C20:1 and cis-7-C20:1) and found that n-3PUFA and LC-MUFA combination significantly decreased lipid accumulation and reduced total cholesterol in HepG2 cells. Cholesterol level was significantly lower in DHA + cis-7-C20:1 group than in DHA + EPA group. These results suggest the importance of LC-MUFA as a functional molecule in fish oil.

Biobased Solvents for Pressurized Liquid Extraction of Nannochloropsis gaditana Omega-3 Lipids.

To develop greener extraction alternatives for microalgae biomass, ultrasound assisted extraction (UAE) and pressurized liquid extraction (PLE) with different biobased solvents were investigated, demonstrating that both techniques are useful alternatives for algal lipid extraction. Specifically, Nannochloropsis gaditana lipids were extracted by UAE and PLE at different temperatures and extraction times with sustainable solvents like 2-Methyltetrahydrofuran (2-MeTHF) and its mixtures with ethanol and other alcohols. The best oil yields for both PLE and UAE of N. gaditana were achieved with the mixture of 2-MeTHF:ethanol (1:3), reaching yields of up to 16.3%, for UAE at 50 °C and up to 46.1% for PLE at 120 °C. Lipid composition of the extracts was analyzed by HPLC-ELSD and by GC-MS to determine lipid species and fatty acid profile, respectively. Different fractionation of lipid species was achieved with PLE and solvent mixtures of different polarity. Thus, for the extraction of glycolipids, ethanolic extracts contained higher amounts of glycolipids and EPA, probably due to the higher polarity of the solvent. The optimized method was applied to microalgae Isochrysis galbana and Tetraselmis chuii showing the potential of mixtures of biobased solvents like 2-methyl-THF and ethanol in different proportions to efficiently extract and fractionate lipids from microalgal biomass.

Beyond Fish Oil Supplementation: The Effects of Alternative Plant Sources of Omega-3 Polyunsaturated Fatty Acids upon Lipid Indexes and Cardiometabolic Biomarkers-An Overview.

Cardiovascular diseases remain a global challenge, and lipid-associated biomarkers can predict cardiovascular events. Extensive research on cardiovascular benefits of omega-3 polyunsaturated fatty acids (n3-PUFAs) is geared towards fish oil supplementation and fish-rich diets. Nevertheless, vegetarianism and veganism are becoming more popular across all segments of society, due to reasons as varied as personal, ethical and religious values, individual preferences and environment-related principles, amongst others. Due to the essentiality of PUFAs, plant sources of n3-PUFAs warrant further consideration. In this review, we have critically appraised the efficacy of plant-derived n3-PUFAs from foodstuffs and supplements upon lipid profile and selected cardiometabolic markers. Walnuts and flaxseed are the most common plant sources of n3-PUFAs, mainly alpha-linolenic acid (ALA), and feature the strongest scientific rationale for applicability into clinical practice. Furthermore, walnuts and flaxseed are sources of fibre, potassium, magnesium, and non-essential substances, including polyphenols and sterols, which in conjunction are known to ameliorate cardiovascular metabolism. ALA levels in rapeseed and soybean oils are only slight when compared to flaxseed oil. Spirulina and Chlorella, biomasses of cyanobacteria and green algae, are important sources of n3-PUFAs; however, their benefits upon cardiometabolic markers are plausibly driven by their antioxidant potential combined with their n3-PUFA content. In humans, ALA is not sufficiently bioconverted into eicosapentaenoic and docosahexaenoic acids. However, evidence suggests that plant sources of ALA are associated with favourable cardiometabolic status. ALA supplementation, or increased consumption of ALA-rich foodstuffs, combined with reduced omega-6 (n6) PUFAs intake, could improve the n3/n6 ratio and improve cardiometabolic and lipid profile.

γ-Linolenic and Stearidonic Acids from Boraginaceae of Diverse Mediterranean Origin.

Thirty Boraginaceae species from different tribes were evaluated in a search of γ-linolenic (GLA, 18:3n-6) and stearidonic acid (SDA, 18:4n-3)-rich oils. The high GLA percentages were found in the seed oils of Symphytum bulbosum and S. tuberosum subsp. tuberosum (27.6 and 27.2 % of total fatty acids (FA)), which are unusually high values for GLA-oils in the current literature. On the whole seed, noticeable GLA percentages were found in S. grandiflorum, S. tuberosum subsp. tuberosum and Borago officinalis (7.43, 4.90, and 4.51 g/100 g, respectively). The main SDA-taxa detected in this study were Buglossoides arvensis, B. incrassata and Glandora oleifolia (21.3, 18.9, and 16.3 % of total fatty acids). On total seed weight, Glandora rosmarinifolia showed the highest SDA content (3.57 g/100 g). Finally, the higher FA contents were found in S. grandiflorum and Paramoltkia doerfleri seeds (35.2 and 37.0 g/100 g, respectively). Principal component analysis showed that similarities in FA profiles allow grouping species as botanical criteria for Boraginaceae tribes do, while the FA groupings confirm the metabolic activities of desaturase and elongase enzymes. Data on the FA composition of the seed oils analyzed here suggest their potential use as functional foods and can be considered as novel sources of SDA and GLA.

Nutraceutical Status and Scientific Strategies for Enhancing Production of Omega-3 Fatty Acids from Microalgae and their Role in Healthcare.

Adherence to Omega-3 fatty acids (O3FAs) as Nutraceuticals for medicinal applications provides health improvement. The prevention and treatment of diseases with O3FAs hold promise in clinical therapy and significantly reduces the risk of chronic disorders. Polyunsaturated fatty acids (PUFA) O3FAs have beneficial effects in the treatment of cardiovascular disorders, diabetic disease, foetal development, Alzheimer's disease, retinal problem, growth and brain development of infants and antitumor effects. Association to current analysis promotes the application of algal biomass for production of O3FAs, mode of action, fate, weight management, immune functions, pharmaceutical and therapeutic applications serving potent sources in healthcare management. A search of the literature was conducted in the databases of WHO website, Sci.org, PubMed, academics and Google. The authors performed search strategies and current scenario of O3FAs in health associated disorders. Promising outcomes and future strategies towards O3FAs may play a pivotal role in Nutraceutical industries in the cure of human health in the future.

Walnut Oil Alleviates Intestinal Inflammation and Restores Intestinal Barrier Function in Mice.

Ulcerative colitis belongs to inflammatory bowel diseases, which is a group of chronic disorders of the gastrointestinal tract. It is a debilitating condition with a wide range of symptoms including rectal bleeding, diarrhea, and visceral pain. Current dietary habits often lead to imbalance in n-6/n-3 polyunsaturated fatty acids (PUFA) in favor of n-6 PUFA. Recent data showed the potential anti-inflammatory advantage of n-3 PUFA. Walnut oil (WO) is rich in those fatty acids and mainly consists of linoleic and linolenic acids that may act via free fatty acids receptors (FFARs). We assessed the anti-inflammatory effect of WO in the mouse model of dextran sulfate sodium (DSS)-induced colitis. Moreover, we examined changes in the expression of tight junction proteins (TJ), pro-inflammatory cytokines, and FFAR proteins in the inflamed mouse colon. WO improves the damage score in inflamed tissue, significantly restoring ion transport and colonic wall permeability. Inflammation caused changes in TJ, FFAR, and pro-inflammatory gene proteins expression, which WO was able to partially reverse. WO has anti-inflammatory properties; however, its exact mechanism of action remains unclear. This stems from the pleiotropic effects of n-3 PUFA ligands associated with receptor distribution and targeted signaling pathways.

Effect on Adipose Tissue of Diabetic Mice Supplemented with n-3 Fatty Acids Extracted from Microalgae.

BACKGROUND: Type 2 Diabetes Mellitus (T2DM) is considered a chronic noncommunicable disease in which oxidative stress is expected as a result of hyperglycaemia. One of the most recent approaches is the study of microalgae fatty acids and their possible antioxidant effect. OBJECTIVE: This study aimed to analyse the effect of supplementation with n-3 fatty acids extracted from microalgae on the total antioxidant capacity (TAC) and lipid peroxidation of adipose tissue and plasma from diabetic (db/db) and healthy (CD1) mice. METHODS: Mice were supplemented with lyophilized n-3 fatty acids extracted from microalgae or added to the diet, from week 8 to 16. TAC assay and Thiobarbituric Acid Reactive Substances assay (TBARS) were performed on adipose tissue and plasma samples. RESULTS: The supplementation of lyophilized n-3 fatty acids from microalgae increased the total antioxidant capacity in adipose tissue of diabetic mice (615.67µM Trolox equivalents vs 405.02µM Trolox equivalents from control mice, p<0.01) and in the plasma of healthy mice (1132.97±85.75µM Trolox equivalents vs 930.64±32µM Trolox equivalents from modified diet mice, p<0.01). There was no significant effect on lipid peroxidation on both strains. CONCLUSION: The use of n-3 fatty acids extracted from microalgae could be a useful strategy to improve total antioxidant capacity in T2DM.

Novel Vibrio spp. Strains Producing Omega-3 Fatty Acids Isolated from Coastal Seawater.

Omega-3 long-chain polyunsaturated fatty acids (LC-PUFAs), such as eicosapentaenoic acid (EPA) (20:5n-3) and docosahexaenoic acid (DHA) (22:6n-3), are considered essential for human health. Microorganisms are the primary producers of omega-3 fatty acids in marine ecosystems, representing a sustainable source of these lipids, as an alternative to the fish industry. Some marine bacteria can produce LC-PUFAs de novo via the Polyunsaturated Fatty Acid (Pfa) synthase/ Polyketide Synthase (PKS) pathway, which does not require desaturation and elongation of saturated fatty acids. Cultivation-independent surveys have revealed that the diversity of microorganisms harboring a molecular marker of the pfa gene cluster (i.e., pfaA-KS domain) is high and their potential distribution in marine systems is widespread, from surface seawater to sediments. However, the isolation of PUFA producers from marine waters has been typically restricted to deep or cold environments. Here, we report a phenotypic and genotypic screening for the identification of omega-3 fatty acid producers in free-living bacterial strains isolated from 5, 500, and 1000 m deep coastal seawater from the Bay of Biscay (Spain). We further measured EPA production in pelagic Vibrio sp. strains collected at the three different depths. Vibrio sp. EPA-producers and non-producers were simultaneously isolated from the same water samples and shared a high percentage of identity in their 16S rRNA genes, supporting the view that the pfa gene cluster can be horizontally transferred. Within a cluster of EPA-producers, we found intraspecific variation in the levels of EPA synthesis for isolates harboring different genetic variants of the pfaA-KS domain. The maximum production of EPA was found in a Vibrio sp. strain isolated from a 1000 m depth (average 4.29% ± 1.07 of total fatty acids at 10 °C, without any optimization of culturing conditions).

Integrated Multitrophic Aquaculture By-Products with Added Value: The Polychaete Sabella spallanzanii and the Seaweed Chaetomorpha linum as Potential Dietary Ingredients.

Aquaculture expansion is limited by the negative environmental impact of the waste and the need for alternative sources in the diet of reared fish. In this framework, for the first time, the survival rates, biomass gain, and fatty acid profiles of the polychaete Sabella spallanzanii and the macroalga Chaetomorpha linum, reared/cultivated as bioremediators in an integrated multitrophic aquaculture system (IMTA), were evaluated for their potential reuse applications. Results showed that these organisms represent a natural source of omega-3 and omega-6. On account of the overall results and the high biomass obtained as by-products, a preliminary study was performed employing both S. spallanzanii and C. linum as new dietary ingredients to feed different sized Dicentrarchus labrax. Fish survival rate, biomass growth, and specific growth rate were determined resulting in no significant differences between control and treated fishes. Histological analyses showed no alterations of the stomach tunica mucosa and submucosa in treated fishes. The eco-friendly approaches applied in the here-realized IMTA system could guarantee the achievement of sustainable by-products represented by the bioremediators S. spallanzanii and C. linum, as well as their reliability as a natural source of compounds beneficial to fish and human health.

Isolation, Characterization and Biotechnological Potentials of Thraustochytrids from Icelandic Waters.

The following study reports on the first thraustochytrid isolates identified from Iceland. They were collected from three different locations off the northern coast of the country (Location A, Skagaströnd; Location B, Hveravík; and Location C, Eyjafjörður). Using 18S rDNA sequence analysis, isolates from Locations A and B were identified within the Thraustochytrium kinnei species while other isolates within the Sicyoidochytrium minutum species when compared to other known strains. Cells isolated from Locations A ( 2 . 10 ± 0 . 70 g/L) and B ( 1 . 54 ± 0 . 17 g/L) produced more biomass than the ones isolated from Location C ( 0 . 43 ± 0 . 02 g/L). This study offers the first-time examination of the utility of byproducts from fisheries as a nitrogen source in media formulation for thraustochytrids. Experiments showed that isolates produced more biomass (per unit of substrate) when cultured on nitrogen of marine ( 2 . 55 ± 0 . 74 g/L) as compared to of commercial origin (  1 . 06 ± 0 . 57 g/L). Glycerol ( 2 . 43 ± 0 . 56 g/L) was a better carbon source than glucose ( 1 . 84 ± 0 . 57 g/L) in growth studies. Fatty acid (FA) profiles showed that the isolates from Location C (S. minutum) had low ratios of monounsaturated ( 4 . 21 ± 2 . 96 % ) and omega-6 ( 0 . 68 ± 0 . 59 % ) FAs. However, the isolates also had high ratios of docosahexaenoic acid (DHA; 35 . 65 ± 1 . 73 % ) and total omega-3 FAs ( 40 . 39 ± 2 . 39 % ), indicating that they could serve as a source of marine oils for human consumption and in aquaculture feeds. The T. kinnei isolates from Location A could be used in biodiesel production due to their high ratios of monounsaturated ( 18 . 38 ± 6 . 27 % ) long chain ( 57 . 43 ± 8 . 27 % ) FAs.

Selective Extraction of ω-3 Fatty Acids from Nannochloropsis sp. Using Supercritical CO2 Extraction.

In this article, microalgae Nannochloropsis sp. was used for fatty acid (FA) extraction, using a supercritical fluid-carbon dioxide (SF-CO2) extraction method. This study investigated the influence of different pre-treatment conditions by varying the grinding speed (200-600 rpm), pre-treatment time (2.5-10 min), and mixing ratio of diatomaceous earth (DE) and Nannochloropsis sp. biomass (0.5-2.0 DE/biomass) on FAs extraction. In addition, the effect of different operating conditions, such as pressure (100-550 bar), temperature (50-75 °C), and CO2 flow rate (7.24 and 14.48 g/min) on eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) recovery, was analyzed. Experimental data evidenced that, keeping constant the extraction conditions, the pre-treatment step enhanced the FAs extraction yield up to 3.4 fold, thereby the maximum extracted amount of FAs (61.19 mg/g) was attained with the pre-treatment with a ratio of DE/biomass of 1 at 600 rpm for 5 min. Moreover, by increasing both SF-CO2 pressure and temperature, the selectivity towards EPA was enhanced, while intermediate pressure and lower pressure promoted DHA recovery. The highest amount of extracted EPA, i.e., 5.69 mg/g, corresponding to 15.59%, was obtained at 75 °C and 550 bar with a CO2 flow rate of 14.48 g/min, while the maximum amount of extracted DHA, i.e., ~0.12 mg/g, equal to 79.63%, was registered at 50 °C and 400 bar with a CO2 flow rate of 14.48 g/min. Moreover, the increased CO2 flow rate from 7.24 to 14.48 g/min enhanced both EPA and DHA recovery.

Tandem Mass Tag Based Quantitative Proteomics of Developing Sea Buckthorn Berries Reveals Candidate Proteins Related to Lipid Metabolism.

Sea buckthorn ( Hippophae L.) is an economically important shrub or small tree distributed in Eurasia. Most of its well-recognized medicinal and nutraceutical products are derived from its berry oil, which is rich in monounsaturated omega-7 (C16:1) fatty acid and polyunsaturated omega-6 (C18:2) and omega-3 (C18:3) fatty acids. In this study, tandem mass tags (TMT)-based quantitative analysis was used to investigate protein profiles of lipid metabolism in sea buckthorn berries harvested 30, 50, and 70 days after flowering. In total, 8626 proteins were identified, 6170 of which were quantified. Deep analysis results for the proteins identified and related pathways revealed initial fatty acid accumulation during whole-berry development. The abundance of most key enzymes involved in fatty acid and triacylglycerol (TAG) biosynthesis peaked at 50 days after flowering, but TAG synthesis through the PDAT (phospholipid: diacylglycerol acyltransferase) pathway mostly occurred early in berry development. In addition, the patterns of proteins involved in lipid metabolism were confirmed by combined quantitative real-time polymerase chain reaction, enzyme-linked immunosorbent assay, and parallel reaction monitoring analyses. Our data on the proteomic spectrum of sea buckthorn berries provide a scientific basic for understanding lipid metabolism and related pathways in the developing berries.

Production of Valuable Compounds and Bioactive Metabolites from By-Products of Fish Discards Using Chemical Processing, Enzymatic Hydrolysis, and Bacterial Fermentation.

The objective of this report was to investigate the isolation and recovery of different biocompounds and bioproducts from wastes (skins and heads) that were obtained from five species discarded by fishing fleets (megrim, hake, boarfish, grenadier, and Atlantic horse mackerel). Based on chemical treatments, enzymatic hydrolysis, and bacterial fermentation, we have isolated and produced gelatinous solutions, oils that are rich in omega-3, fish protein hydrolysates (FPHs) with antioxidant and antihypertensive activities, and peptones. FPHs showed degrees of hydrolysis higher than 13%, with soluble protein concentrations greater than 27 g/L and in vitro digestibilities superior to 90%. Additionally, amino acids compositions were always valuable and bioactivities were, in some cases, remarkable. Peptones that were obtained from FPHs of skin and the heads were demonstrated to be a viable alternative to expensive commercial ones indicated for the production of biomass, lactic acid, and pediocin SA-1 from Pediococcus acidilactici.