Enrichment Of Polyunsaturated Fatty Acids Research Articles

Direct Comparison of 18 Carbon n–3 and n–6 Fatty Acids at Equal Levels in an Oil Blend on Tissue Enrichment of Long-Chain Polyunsaturated Fatty Acid in Broiler Chickens

BackgroundOmega-3 (n-3) and omega-6 (n-6) polyunsaturated fatty acids (PUFAs) are of interest because of their health effects. However, most experiments use natural oils and are confounded by PUFA concentrations and other fatty acids (FAs) that impact biosynthesis of the very long-chain derivatives (VLC). ObjectivesTo directly compare the effect of 18 C n-3 or n-6 FA fed at similar rates on their elongation and desaturation to VLC PUFA and their incorporation into tissues. MethodsOil blends that substituted ∼23% points of stearidonic acid (SDA) with alpha-linolenic acid (ALA), gamma-linolenic acid (GLA), or linoleic acid (LA) while minimizing differences in other FA were prepared. COBB500 broilers were fed the oil blends at 1.25% of the diet from day 14–35 age. ResultsThere was greater enrichment of VLC PUFA in breast, thigh, liver, and plasma when diets were supplemented with high-SDA and high-GLA oil blends than high-ALA and high-LA oil blends. The efficiency of VLCn-3 PUFA synthesis from SDA and ALA was lower than the efficiency of VLCn-6 PUFA synthesis from GLA and LA, suggesting that the elongation and desaturation enzymes more efficiently utilized n-6 substrates. The efficiency of biotransformation of SDA to VLCn-3 PUFA was greater than that of high-ALA, and synthesis of VLCn-6 PUFA from GLA was higher than that of high-LA in breast, thigh, liver, and plasma. There were minimal effects on tissue-saturated and monounsaturated FA. ConclusionsThe high-SDA and high-GLA oil blends efficiently enriched tissues with their VLC-PUFA more than high-ALA and high-LA treatments.

Characterization of a New Thermostable and Organic Solution-Tolerant Lipase from Pseudomonas fluorescens and Its Application in the Enrichment of Polyunsaturated Fatty Acids.

The search for and characterization of new lipases with excellent properties has always been urgent and is of great importance to meet industrial needs. In this study, a new lipase, lipB, from Pseudomonas fluorescens SBW25, belonging to the lipase subfamily I.3, was cloned and expressed in Bacillus subtilis WB800N. Enzymatic properties studies of recombinant LipB found that it exhibited the highest activity towards p-nitrophenyl caprylate at 40 °C and pH 8.0, retaining 73% of its original activity after incubation at 70 °C for 6 h. In addition, Ca2+, Mg2+, and Ba2+ strongly enhanced the activity of LipB, while Cu2+, Zn2+, Mn2+, and CTAB showed an inhibiting effect. The LipB also displayed noticeable tolerance to organic solvents, especially acetonitrile, isopropanol, acetone, and DMSO. Moreover, LipB was applied to the enrichment of polyunsaturated fatty acids from fish oil. After hydrolyzing for 24 h, it could increase the contents of polyunsaturated fatty acids from 43.16% to 72.18%, consisting of 5.75% eicosapentaenoic acid, 19.57% docosapentaenoic acid, and 46.86% docosahexaenoic acid, respectively. The properties of LipB render it great potential in industrial applications, especially in health food production.

Endogenous production of ω-3 polyunsaturated fatty acids mitigates cisplatin-induced myelosuppression by regulating NRF2-MDM2-p53 signaling pathway.

Cisplatin is a chemotherapy medication used to treat a wide range of cancers. A common side effect of cisplatin is myelosuppression. Research suggests that oxidative damages are strongly and consistently related to myelosuppression during cisplatin treatment. ω-3 polyunsaturated fatty acids (PUFAs) can enhance the antioxidant capacity of cells. Herein, we investigated the protective benefit of endogenous ω-3 PUFAs on cisplatin-induced myelosuppression and the underlying signaling pathways using a transgenic mfat-1 mouse model. The expression of mfat-1 gene can increase endogenous levels of ω-3 PUFAs by enzymatically converting ω-6 PUFAs. Cisplatin treatment reduced peripheral blood cells and bone marrow nucleated cells, induced DNA damage, increased the production of reactive oxygen species, and activated p53-mediated apoptosis in bone marrow (BM) cells of wild-type mice. In the transgenics, the elevated tissue ω-3 PUFAs rendered a robust preventative effect on these cisplatin-induced damages. Importantly, we identified that the activation of NRF2 by ω-3 PUFAs could trigger an antioxidant response and inhibit p53-mediated apoptosis by increasing the expression of MDM2 in BM cells. Thus, endogenous ω-3 PUFAs enrichment can strongly prevent cisplatin-induced myelosuppression by inhibiting oxidative damage and regulating the NRF2-MDM2-p53 signaling pathway. Elevation of tissue ω-3 PUFAs may represent a promising treatment strategy to prevent the side effects of cisplatin.

Microalgal polyunsaturated fatty acids: Hotspots and production techniques.

Algae play a crucial role in the earth's primary productivity by producing not only oxygen but also a variety of high-value nutrients. One such nutrient is polyunsaturated fatty acids (PUFAs), which are accumulated in many algae and can be consumed by animals through the food chain and eventually by humans. Omega-3 and omega-6 PUFAs are essential nutrients for human and animal health. However, compared with plants and aquatic sourced PUFA, the production of PUFA-rich oil from microalgae is still in the early stages of exploration. This study has collected recent reports on algae-based PUFA production and analyzed related research hotspots and directions, including algae cultivation, lipids extraction, lipids purification, and PUFA enrichment processes. The entire technological process for the extraction, purification and enrichment of PUFA oils from algae is systemically summarized in this review, providing important guidance and technical reference for scientific research and industrialization of algae-based PUFA production.

P068 Polyunsaturated fatty acid excess fuels pro-inflammatory transcriptional networks in GPX4 deficient intestinal epithelial cells

Abstract Background There is increasing evidence that the rise in Crohn’s disease (CD) incidence is partially driven by constituents of a Western-style diet such as polyunsaturated fatty acids (PUFA). Recently we could demonstrate that PUFA excess induces a CD-like enteritis in mice lacking antioxidant GPX4 in their intestinal epithelial cells (IECs) and that CD patients clinical disease activity positively correlates with PUFA intake. In this project we investigated the role of epithelial lipid composition and lipid mediators during PUFA induced metabolic gut inflammation. Furthermore, we transcriptionally profiled GPX4 deficient small intestinal cells after stimulation with ω3- and ω6-PUFA. Methods Mice lacking one allele of Gpx4 in their IECs (GPX4+/-IEC) and wild-type controls (WT) were fed a PUFA (10% fish oil) enriched Western diet (PUFA-WD) for 12 weeks. Intestinal inflammation was assessed by histopathology. IECs were harvested by scraping of small intestinal sections and lipid profiles and mediators were analysed with LC/MS. GPX4 in murine small intestinal cells (MODE-K) was knocked down by siRNA (siGpx4) scrambled siRNA served as control (siCtrl). MODE-K were stimulated with ω3-, ω6-PUFA, 5,6- and 8,9-DHET or vehicle. Cytokine expression in cell culture supernatant was analysed by ELISA and transcriptome was analysed by bulk RNA sequencing. Results Histopathology revealed gut inflammation in GPX4+/-IEC but not WT mice after 12 weeks PUFA-WD feeding. LC/MS analysis of IECs derived from GPX4+/-IEC and WT mice fed the PUFA-WD depicted a significant increase in eicosapentaenoic acid (20:5), docosapentaenoic acid (22:5) and docosahexaenoic acid (22:6) containing phosphatidylethanolamine (PE) and phosphatidylcholine (PC) species (Fig 1a). All other PE and PC species showed a reduced amount, confirming the enrichment of the specific PUFAs within the IECs by the PUFA-WD. Lipid mediator analysis further revealed an increase in 5,6-DHET and 8,9-DHET only in IECs derived from GPX4+/-IEC but not WT mice (Fig 1b). When stimulated with PUFA, siGpx4 MODE-K produced high amounts of neutrophil attracting chemokine CXCL-1 which is attenuated by the supplementation of anti-inflammatory lipid mediators 5,6-DHET and 8,9-DHET (Fig 2). Transcriptome analysis revealed the upregulation of pro-inflammatory pathways including TNFa, MAPK and NF-κB in siGpx4 vs. siCtrl MODE-K after PUFA stimulation (Fig 3). Conclusion By integrating lipidomic and transcriptomic data of a reductionist cell culture system and a model organism, we could show that PUFA excess in a GPX4 deficient individual fuels a pro-inflammatory transcriptional network which outweighs anti-inflammatory ω3-PUFA metabolites influence culminating in gut inflammation.

Correction of the fatty acid composition of purpose dietary fats

It has been indicated that, according to very numerous studies, substantial indicators of the functionality of fatty products are the ratio of polyunsaturated fatty acids (PUFAs) of ω6 and ω3 families, as well as the ratio of the sum of saturated, monounsaturated and polyunsaturated fatty acids (SFA:MUFA:PUFA). Points related to the fortification of the functional properties of fat products of a target (special) purpose by combining their composition with the use of crushed non-defatted (full-fat) masses of oily flax and sesame seeds have been considered. The biological potential of the crushed non-defatted mass of oil flax seeds as a promising component for the enrichment of PUFA of ω3 group of new food vegetable-fat composite mixtures for special purposes have been theoretically substantiated. Insofar as margarines, which are often not balanced in terms of the ratio of fatty acids ω6 and ω3, are usually included as a fat component in the recipes of many bakery and flour confectionery products, this article proposes pre-mixing margarine with non-defatted crushed mass of oil flax and sesame seeds, without extracting the oil from their composition, before adding it to the dough.

Feeding laying hens docosa hexaenoic acid-rich microalgae oil at 40 g/kg diet causes hypotriglyceridemia, depresses egg production, and attenuates expression of key genes affecting hepatic triglyceride synthesis and secretion, but is rescued by dietary co-supplementation of high-oleic sunflower oil

The primary goal of this study was to investigate the effect of feeding White Leghorn hens graded levels of a docosahexaenoic acid (DHA)-rich microalgae oil (MAO) on productive performance and enrichment of eggs with very long-chain (VLC) omega-3 (n-3) polyunsaturated fatty acids (PUFA). Forty-nine-week-old hens (8 per diet) were fed the following diets for 28 d: 1) A corn-soybean meal-based diet with no supplemental oil (CON); 2) CON+10 g/kg MAO; 3) CON+20 g/kg MAO; 4) CON+30 g/kg MAO; 5) CON+40 g/kg MAO; 6) CON+40 g/kg MAO+20 g/kg high-oleic sunflower oil (HOSO); and 7) CON+40 g/kg MAO+40 g/kg HOSO. Diets 6 and 7 were included because we previously reported that co-feeding high-oleic acid oils with n-3 PUFA-containing oils attenuated egg yolk n-3 PUFA contents vs. feeding hens the n-3 oils alone. All data were collected on an individual hen basis. Egg VLC n-3 PUFA enrichment plateaued, in terms of statistical significance, at the 30 g/kg MAO level (266 mg/yolk). Hens fed 40 g/kg MAO had greatly attenuated measures of hen performance, marked liver enlargement, an altered ovarian follicle hierarchy, greatly lowered circulating triglyceride levels, and depressed hepatic expression of key genes involved in triglyceride synthesis and secretion. As compared to hens fed 40 g/kg MAO alone, feeding hens 40 g/kg MAO co-supplemented with HOSO (Diets 6 and 7) restored egg production, ovarian morphology, and all other measures of hen productive performance to CON levels, elevated plasma triglyceride levels, prevented liver enlargement, and increased the hepatic expression of key genes involved in triglyceride synthesis and secretion. In conclusion, MAO can greatly enrich hens' eggs with VLC n-3 PUFA, but its recommended dietary inclusion should not exceed 20 g/kg. This would allow for near-maximal yolk VLC n-3 PUFA enrichment without impairing hen productive performance, altering the ovarian follicle hierarchy or, based on the work of others, presumably imparting off-flavors in the egg.

Exposure to the Brominated Flame Retardant Hexabromocyclododecane Results in Systemic Changes in Polyunsaturated Fatty Acid Metabolism Within Phospholipids of Mice

Brominated flame retardants are used in many household products to reduce flammability, but often leach into the surrounding environment over time. Hexabromocyclododecane (HBCD) is one brominated flame retardant detected in human blood across the world. HBCD exposure can result in neurological problems and altered lipid metabolism, but to date the two remain unlinked. As lipids constitute ~50% of brain dry weight, lipid metabolism plays a critical role in neuronal function and homeostasis. To determine the effect of HBCD exposure on brain lipid metabolism, young adult male mice were exposed to 1 mg/kg HBCD every 3 days for 28 days. HBCD exposure altered lipid abundance of 8 major lipid classes in the dorsal striatum, including fatty acids (FA), lysophosphatidylcholines (LPC), lysophosphatidylethanolamines (LPE), phosphatidylcholines (PC), phosphatidylethanolamines (PE) alkylphosphatidylethanolamines (PEO), phosphatidylethanolamine plasmalogens (PEP), and sphingomyelins (SM). The liver, as a major lipid metabolism hub, and blood were also examined to determine whether these alterations correlated to systemic changes in lipid metabolism. Although several lipid classes changed in the liver and blood, only PE was consistently altered between organs. Despite the lack of commonalities between lipid class alterations of differing organs, phospholipid fatty acid tails displayed a consistent enrichment of Polyunsaturated Fatty Acids (PUFAs) such as Arachidonic Acid (AA) and Docosahexaenoic Acid (DHA) across lipid classes and organs. Free AA was enriched in the blood as well as AA contained within phospholipids of the liver (in PE), blood (in LPC, PEO, and PEP), and dorsal striatum (in PE, PEO). Additionally, although free DHA was unchanged in the blood, DHA‐containing phospholipids were enriched in the liver (in PE), blood (in LPC, LPE), and dorsal striatum (in PE, PEO, PEP, PS). These studies indicate that HBCD can cause systemic alterations in PUFA metabolism. Further study is necessary to determine the mechanisms underlying the lipid alterations and their functional consequences.

Application of undecanedicarboxylic acid to prepare cross-linked enzymes (CLEs) of Rhizomucor miehei lipase (RML); Selective enrichment of polyunsaturated fatty acids

We have introduced 1,11-undecanedicarboxylic acid as a new cross-linker to prepare cross-linked preparation of Rhizomucor miehei lipase (RML) from its soluble form by applying an isocyanide-based multi-component reaction. This one-step cross-linking of soluble enzyme allowed skipping the prior step of enzyme aggregation or crystallization which is inevitable in the traditional methods of cross-linking. High immobilization yields were achieved up to 92% in an extremely mild condition. The addition of bovine serum albumin (BSA) as a feeder protein affected the final properties of RML by producing 2.5-fold higher specific activity compared to the specific activity of free RML. Furthermore, dynamic light scattering analysis (DLS) showed that by applying BSA, the cross-linked enzymes (CLEs) particle sizes of 1.75–3.91 µm were obtained. Considerable improvement in the thermal and co-solvent stability was observed, in particular for the derivative obtained by immobilization in the presence of BSA. This derivative remained considerably active at 60 °C by keeping 79% of its initial activity after 2 h of incubation and retained 75% and 86% of its activity after 24 h incubation in the presence of 20% of methanol and ethanol, respectively. The cross-linked preparations were then applied for the selective hydrolysis of the essential omega-3 fatty acids, including eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), from fish oil. The results showed that the immobilized derivatives tend to preferentially hydrolyze EPA.

Utilization of Live‐Food Enrichment with Polyunsaturated Fatty Acids (PUFA) for the Intensive Culture of Yellow Perch Larvae

AbstractChallenges associated with first feeding have impeded the intensive culture of larval Yellow Perch Perca flavescens. Live‐food enrichment with polyunsaturated fatty acids (PUFAs) can increase the growth and survival of larval fish, but this method has not been tested with Yellow Perch. This study includes two experiments that were meant to evaluate (1) the relative proportion of docosahexaenoic acid (DHA; 22:6[n‐3]) and arachidonic acid (ARA; 20:4[n‐6]) in enrichment emulsions and (2) the provision of PUFA emulsions in ethyl ester (EE) or triacylglycerol (TG) form on Yellow Perch growth and survival. Fish were provided with live rotifers Brachionus plicatilis and brine shrimp Artemia nauplii for the first 10 d of exogenous feeding within a specialized recirculating system (phase I). Fish were then transferred to flow‐through tanks and were fed Artemia nauplii for 3 d before gradually transitioning to a formulated starter diet (5–7 d; phase II). Fish size, growth, survival, swim bladder inflation rates, and lipid/fatty acid concentrations were evaluated after each phase. Overall, rotifers assimilated ARA in higher concentrations (23.3% of total fatty acids) than Artemia (0.6–0.7% of total fatty acids). Rotifers also tended to assimilate PUFAs better in EE form than in TG form (71.3% versus 66.5% Σn‐3 + n‐6 fatty acids). In the first experiment, fish from the ARA‐ and DHA‐enriched diet groups had greater swim bladder inflation rates (phase I) and growth rates (phase II) than the unenriched control group. In phase I of the second experiment, the EE enrichment group had a significantly faster growth rate than the TG enrichment group (specific growth rate: mean ± SD = 40.5 ± 0.9% and 36.6 ± 1.8%, respectively). Fatty acid composition of zooplankton was heavily influenced by enrichments, and fatty acid composition of larvae/juveniles reflected that of their live prey. The results of this study suggest that PUFA enrichment of live feeds can accelerate Yellow Perch growth and reduce the time spent in the critical period of early development.

Enrichment of polyunsaturated fatty acids in black soldier fly larvae (Hermetia illucens) fortified with squid liver oil

Black soldier fly larvae (BSFL;Hermetia illucens) are known as an alternative feed for livestock, but their lack of polyunsaturated fatty acids (such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)) makes it less valuable. To overcome these problems, BSFL substrate (chicken feed) was fortified with squid liver oil (SLO) at five different concentrations (0, 2.5, 5, 10, and 20%). The growth rate, feed conversion, and nutritional content of BSFL were evaluated at day 15 of rearing. Of the five concentrations, SLO 5% showed the highest growth increase (25.82-fold) among the other treatments (20.63-22.98-fold;P<0.05). The fortification of SLO 5% did not result in differences in survival and feed conversion of larvae compared to larvae fed the control substrate. By rearing BSFL in a substrate containing SLO 5%, the lipid content of the larvae was 32% higher than that of larvae fed the control substrate (P<0.05). The accumulation of lipids was faster in the substrate containing SLO 5% (33.20% for 8 days) than in the control substrate (24.36% for 15 days). The control group of BSFL contained no DHA or EPA, but after rearing on the SLO 5% fortified substrate, the larvae contained DHA at an average level of 2.99 g/100 g lipid and EPA was 2.68 g/100 g lipid. Harvested larvae from SLO 5% treatment was within safe levels of Pb, Cd, As, and Hg (840, 370, 860, and 26.7 μg/kg, respectively), under the EU threshold for animal feed. Based on our results, it concluded that BSFL enriched with PUFAs, DHA and EPA can be considered as important nutritional components of animal feed without excessive heavy metals accumulation by feeding SLO in an appropriate amount.

Screening the rate-limiting genes in the ω6 polyunsaturated fatty acid biosynthesis pathway in Nannochloropsis oceanica

The enrichment of polyunsaturated fatty acids has made Nannochloropsis sp. a promising candidate for the production of polyunsaturated fatty acids. Although most of the genes involved in polyunsaturated fatty acid biosynthesis have been cloned, the regulatory mechanism at the molecular level has not been elucidated. To target the rate-limiting genes in the ω6 polyunsaturated fatty acid biosynthesis pathway in Nannochloropsis oceanica, we performed the following experiments. The function of the Δ9 fatty acid desaturase gene of N. oceanica was verified in Saccharomyces cerevisiae with a substrate preference for stearic acid. With gas chromatography–mass spectrometry (GC–MS) analysis, the fatty acid profiles in N. oceanica under different stress conditions (high nitrogen, low nitrogen, high temperature, and low temperature) were measured. The results showed that high nitrogen and low temperature stresses played significant roles in promoting polyunsaturated fatty acid biosynthesis. In contrast, low nitrogen and high temperature stresses were beneficial for the accumulation of monounsaturated fatty acids. We further measured the transcriptional abundances of all six genes in the ω6 polyunsaturated fatty acid biosynthesis pathway in N. oceanica and found that the Δ6 fatty acid elongase and the Δ12, Δ5, and Δ17 fatty acid desaturase genes were more sensitive to the stresses set in this study. Pearson correlation analysis showed that the Δ9 fatty acid desaturase was significantly associated with monounsaturated fatty acid biosynthesis, while Δ12, Δ5, and Δ17 fatty acid desaturases were proposed to be the rate-limiting enzymes in polyunsaturated fatty acid biosynthesis. This study helps to thoroughly elucidate the responses of N. oceanica to nitrogen and temperature stresses and optimize cultivation for high yield and quality of polyunsaturated fatty acids. More importantly, the selected rate-limiting genes may benefit further genetic engineering in N. oceanica.

A comparative study of fatty acids profile of two Indian major carps (Gibelion catla, Hamilton, 1822 and Cirrhinus mrigala, Hamilton, 1822) using value added fish feed

Freshwater fishes are not only a major source of protein but they also possess nutritionally valuable lipids in the form of Polyunsaturated fatty acids (PUFAs), which play a crucial role in the normal growth, disease prevention, development, cardiovascular health and reproduction of human. The present study was performed to determine the incorporation rate of fatty acids profile and their composition in two common freshwater carps as Gibelion catla and Cirrhinus mrigala (in situ trial and experimental) in the different experimental time period (0 days, i.e. initial, 90 days and 180 days) by using of value added feed like flaxseed (?-linolenic acids, 51.26% – 54.94%) and soybean oil (?-linolenic acids, 7.95%-9.01%) as omega-3 supplements. To determine the specific growth pattern Length-Weight Relationships (LWRs) are analyzed where it showed positive allometric growth (b=3.20 in 90 days, b=3.11 in 180 days for Catla and b=3.18 in 90 days, b=3.1 in 180 days for Mrigala fish). The Gas Chromatography-Mass Spectrometry (GC/MS) method also confirmed that the percentages of EPA (eicosapentaenoic acid) and DHA (docosahexaenoic acid) increased significantly (P< 0.05) in experimental (0.096a±0.41, initial; 5.16a±0.27, 90 days; 6.21b±0.36, 180 days Catla fish species and 0.019a±0.96 initial; 3.74b±0.37, 90 days; 3.50a±0.46 180 days for Mrigal fish species) fishes rather than controls (4.28a±0.27, 90 days; 4.36b±0.36, 180 days for Catla species and 2.24b±0.31 90 days; 2.50a±0.11 180 days for Mrigal species). Therefore, it was clearly indicated that formulated diet performed significantly to maintain the positive allometric growth as well as successive enrichment of PUFAs in experimental specimens, which is beneficial for human health as high source of protein and PUFAs as well.

Effect of lowering the amount of dietary linoleic acid on tissue omega-3 fatty acid contents of broilers fed supplemental flaxseed oil from 18 to 35 days of age

Chickens can hepatically synthesize very long-chain omega-3 ( VLC n-3 ) polyunsaturated fatty acids ( PUFA ) from α-linolenic acid ( ALA ; 18:3 n-3); however, the efficiency is attenuated by linoleic acid ( LA ; 18:2 n-6), which competes with ALA for the same elongase and desaturase enzymes. Here, we fed broilers supplemental ALA-rich flaxseed oil ( FLAX ) while reducing the dietary level of LA to determine if greater tissue n-3 PUFA enrichment could be attained. Male Ross 708 chicks were fed a common starter diet from day 1 to 17. On day 18, 4 pens of 36 birds were each fed one of 5 diets: (1) reduced-LA/low-n-3 fatty acid base + 60 g/kg high-oleic sunflower oil ( HOSO ); (2) reduced-LA base + 50 g/kg FLAX+ 10 g/kg HOSO; (3) standard-LA base + 50 g/kg FLAX + 10 g/kg HOSO; (4) standard-LA base + 50 g/kg FLAX + 10 g/kg high-LA safflower oil; and (5) standard-LA base + 50 g/kg FLAX + 10 g/kg fish oil. There was no effect ( P > 0.05) of diet on day-35 BW, FCR, breast, thigh, or abdominal fat pad weights, while birds fed diet 2 had higher thigh ALA, total n-3, and total VLC n-3 PUFA than birds fed diets 1, 3, and 4. Breast data followed a similar pattern, although the differences were not significant ( P > 0.05). In conclusion, reducing the LA content of a FLAX-supplemented diet promoted greater deposition of n-3 PUFA in broiler muscles, although the highest tissue contents of VLC n-3 PUFA were observed in birds fed fish oil (diet 5).

Enrichment of Polyunsaturated fatty acids from catfish oil with immobilized Rhizopus oryzea lipase

Nowadays, Pangasius hypophthalmus fillet is one of the main marine products in Vietnamese Mekong River Delta with over three hundred thousand tons per year. One of the byproducts of this industry is fat, about one hundred thousand tons per year. That byproduct has been only used in animal foodstuff industry after oil extracting or it becomes waste. In order to increase the quality of Pangasius hypophthalmus oil, we have enriched PUFA in the oil by hydrolysis with immobilized Rhizopus oryzea lipase. Lipase has been immobilized on celite by absorption method. Immobilization experiments have been designed in a two-level full factorial model with three factors. The fish oil has been hydrolyzed with immobilized lipase to enrich PUFA with optimized parameters found out: the ratio
 of enzyme to support of 1:10 g/g, the ratio of phosphate buffer volume to support weight of 30.55 ml/g, incubation time of 5.15 hours. The result has showed that 60-minute hydrolysis is the most appropriate. Additionally, the oil iodine value has increased from 71.4 to 97.4, and DHA concentration has risen from 1156mg/kg to 1916mg/kg.

Abstract B48: Ovarian cancer risk in laying hens is reduced by dietary polyunsaturated fatty acids: Implications for soluble E-cadherin, de novo lipogenesis, and mitochondrial metabolism

Abstract Aim: The specific aim of our research is to understand the relationship between soluble E-cadherin (sE-cad) expression, de novo lipogenesis, and mitochondrial metabolism in ovarian tumors from White Leghorn laying hens, in response to enrichment with diets that contain high levels of polyunsaturated fatty acids (PUFAs). Background: The shedding of the 80 kd extracellular domain of E-cadherin (E-cad) causes the production of sE-cad, which is implicated as a putative ligand for growth, survival, and angiogenesis in solid tumors. Little is known with regard to the relationship between sE-cad production and de novo lipogenesis or mitochondrial metabolism in ovarian tumors. De novo lipogenesis has been argued to be indispensable for their growth and proliferation. Vast physiologic research indicates that de novo lipogenesis is inhibited by PUFAs, indicating that dietary PUFAs could provide negative feedback to an essential metabolic pathway in ovarian tumors. Epidemiologic studies of human populations indicate that ovarian cancer (OvCa) risk is reduced 30-40% by high dietary consumption of omega-3 or omega-6 PUFAs. Hypothesis: We hypothesize that high dietary PUFA consumption downregulates de novo lipogenesis in ovarian tumors of laying hens, correlating with reduced sE-cad production and increased mitochondrial oxidative phosphorylation (OXPHOS) capacity. Methods: We used qPCR to measure genes related to de novo lipogenesis and mitochondrial OXPHOS. Gas chromatography was used measure the fatty acid content in ovarian tumors. Western blot was used to measure E-cad protein fragments. Results: Our lab observed a 20-40% reduction of OvCa risk in laying hens that were supplemented with whole flaxseed, flaxseed oil, corn oil or fish oil. qPCR results indicate reduced expression of genes related to de novo lipogenesis (i.e., Fatty Acid Synthase). Gas chromatography results indicate that the content of C16:0, C18:0 and C18:1 fatty acids is similarly reduced by dietary PUFA enrichment. qPCR results also indicate increased expression of the catalytic subunits for complexes 3, 4, and 5 of the mitochondrial electron transport chain when diets are enriched with PUFAs. Lastly, although the expression of E-cad protein was unaffected by diet, we did detect a significant reduction of the 37 kd carboxy-terminal fragment of E-cad (CTF1-Ecad) in the fish oil diet. This is an important finding, because CTF1-E-cad is the complementary fragment of sE-cad. Conclusions: OvCa risk in hens is reduced by consumption of omega-3 and omega-6 PUFAs, and these risk reductions associate with reduced de novo lipogenesis, reduced CTF1-E-cad expression, and improved mitochondrial OXPHOS capacity. This study reveals an additional anti-ovarian cancer mechanism of a flaxseed-supplemented diet. (NIH-RO1AT005295) Citation Format: Chris Weston, Matthew Young, Karen Hales, Dale Buchanan Hales. Ovarian cancer risk in laying hens is reduced by dietary polyunsaturated fatty acids: Implications for soluble E-cadherin, de novo lipogenesis, and mitochondrial metabolism [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research; 2019 Sep 13-16, 2019; Atlanta, GA. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(13_Suppl):Abstract nr B48.

A transferase interactome that may facilitate channeling of polyunsaturated fatty acid moieties from phosphatidylcholine to triacylglycerol

Polyunsaturated fatty acids (PUFAs) such as α-linolenic acid (ALA, 18:3Δ9cis,12cis,15cis ) have high nutritional and industrial values. In oilseed crops, PUFAs are synthesized on phosphatidylcholine (PC) and accumulated in triacylglycerol (TAG). Therefore, exploring the mechanisms that route PC-derived PUFA to TAG is essential for understanding and improving PUFA production. The seed oil of flax (Linum usitatissimum) is enriched in ALA, and this plant has many lipid biosynthetic enzymes that prefer ALA-containing substrates. In this study, using membrane yeast two-hybrid and bimolecular fluorescence complementation assays, we probed recombinant flax transferase enzymes, previously shown to contribute to PUFA enrichment of TAG, for physical interactions with each other under in vivo conditions. We found that diacylglycerol acyltransferases, which catalyze the final reaction in acyl-CoA-dependent TAG biosynthesis, interact with the acyl-editing enzymes phosphatidylcholine: diacylglycerol cholinephosphotransferase, and lysophosphatidylcholine acyltransferase. Physical interactions among the acyl-editing enzymes were also identified. These findings reveal the presence of an assembly of interacting transferases that may facilitate the channeling of PUFA from PC to TAG in flax and possibly also in other oleaginous plants that produce seeds enriched in PC-modified fatty acids.

Ovarian Cancer Risk in Laying Hens Is Reduced by Dietary Polyunsaturated Fatty Acids: A Case for Redox Homeostasis and Reprogrammed Mitochondrial Metabolism in Ovarian Tumors (OR04-05-19)

ObjectivesIn 2012, ovarian cancer accounted for 239,000 new cancer cases and 152,000 cancer-related deaths worldwide. Epidemiological studies around the world suggest that high dietary consumption of polyunsaturated fatty acids (PUFAs) reduces a woman’s odds of developing ovarian cancer. We hypothesize that PUFA-enriched diets will decrease ovarian cancer risk in White Leghorn laying hens. MethodsThe specific aim of our research was to investigate ovarian cancer risk and molecular events in ovarian tumors of White Leghorn laying hens consuming PUFA-enriched diets. We exposed laying hens to long-term (46-week) dietary PUFA treatment, using Whole Flaxseed (WFX), Flaxseed Oil (FXO), Fish Oil (FHO), Corn Oil (CNO), and a low-PUFA diet, Defatted Flaxseed (DFX). At the end of the dietary treatment we collected 88 ovarian tumors and conducted qPCR analysis for gene expression. ResultsIn comparison to a control diet lacking a PUFA supplement, the odds ratios of hens developing ovarian cancer decreased with PUFA enrichment (Odd Ratios: DFX = 0.74, WFX = 0.69, FXO = 0.61, FHO = 0.57, CNO = 0.57), corroborating human epidemiological observations of ovarian cancer risk. Gene expression analysis from hen ovarian tumors suggests that PUFA-rich diets (ie WFX, FXO, FHO, and CNO) improve mitochondrial electron transport chain function by upregulating Cytochrome B (Complex 3) and Cyctochrome C Oxidases 1, 2, and 3 (Complex 4), and likely reduce superoxide emission from Complex 1 as evidenced by the downregulation of NADH Dehydrogenase 2. PUFA enrichment also reduced the expression of enzymes associated with reactive oxygen species (ROS), reactive nitrogen species (RNS), and lipid peroxides, suggesting overall major improvement in the redox state of tumors from PUFA-enriched diets. The FXO diet was uniquely distinguished by increasing the expression of ATP5B1 (catalytic subunit of ATP Synthase) and the ADP/ATP Translocase (ANT1), suggesting increased mitochondrial cristae space and improved oxidative phosphorylation capacity in FXO-enriched ovarian tumors. ConclusionsDietary PUFA enrichment (omega-3 or omega-6) reduces end point risk for ovarian cancer in laying hens. The underlying etiology appears to center on decreasing oxidative stress and increasing mitochondrial function in ovarian tumors. Funding SourcesNational Institutes of Health.

N-3 polyunsaturated fatty acids: An innovative strategy against obesity and related metabolic disorders, intestinal alteration and gut microbiota dysbiosis

Obesity is now widely recognized to be associated with low-grade systemic inflammation. It has been shown that high-fat feeding modulates gut microbiota which strongly increased intestinal permeability leading to lipopolysaccharide absorption causing metabolic endotoxemia that triggers inflammation and metabolic disorders. N-3 polyunsaturated fatty acids (PUFAs) have been shown associated with anti-obesity properties, but results still remain heterogeneous and very few studies underlined the metabolic pathways involved. Thus, the use of Fat-1 transgenic mice allows to better understanding whether endogenous n-3 PUFAs enrichment contributes to obesity and associated metabolic disorders prevention. It specially evidence that such effects occur through modulations of gut microbiota and intestinal permeability. Then, by remodeling gut microbiota, endogenous n-3 PUFAs improve HF/HS-diet induced features of the metabolic syndrome which in turn affects host metabolism. Thus, increasing anti-obesogenic microbial species in the gut microbiota population (i.e Akkermansia) by appropriate n-3 PUFAs may represent a promising strategy to control or prevent metabolic diseases.

Lipase-Mediated Selective Methanolysis of Fish Oil for Biodiesel Production and Polyunsaturated Fatty Acid Enrichment

Fish oil, containing quite a large amount of polyunsaturated fatty acids (PUFAs), is well-recognized as a good source for biodiesel and health care product production. A two-step process involving free lipase NS81006-mediated methanolysis followed by immobilized lipase Novozym 435-catalyzed conversion was proposed for the utilization of fish oil in this paper. During the lipase NS81006-catalzyed methanolysis process, the selectivity of lipase on different free fatty acids (FFAs) was studied systematically, and it was found that the length of the carbon chain and the number and position of C–C double bonds had a varied influence on the selectivity of lipase. The reaction rate and biodiesel yield decreased with the increase of the carbon chain length; higher conversion could be obtained with unsaturated FFAs (C18:1, C18:2, and C18:3) compared to unsaturated FFA (C18:0), among which FFA with three double bonds gave the highest conversion. Through this two-step lipase-mediated catalysis, both the conventional...