Upgrading of maatjes herring byproducts: production of crude fish oil

Fish oil has been extracted from byproducts of the maatjes (salted) herring production using a pilot plant consisting of a mincer, heat exchanger, and three-phase decanter. The crude herring oil obtained had an initial peroxide value (PV), anisidine value (AV) and free fatty acids (FFA) level of only 3 mequiv of peroxide/kg of lipid, 8.9, and 2.9%, respectively. 5,8,11,14,17-Eicosapentaenoic acid (EPA) and 4,7,10,13,16,19-docosahexaenoic acid (DHA) were present in considerable amounts (99 and 91 g/kg, respectively). During storage of the oil, no photooxidation could be detected. Storage at room temperature led to significant autoxidation over time, apparent from primary and tertiary oxidation products, measured by a decrease of hydroperoxides and an increase of fluorescent compounds (FC). Storage at 50 degrees C resulted in significant increases in secondary (AV) and tertiary oxidation (FC) products. At all storage conditions, the FFA contents remained low (<3%) and the alpha-tocopherol content remained constant. These results open the possibility for fish oil production of good quality using salted herring byproducts.     

Lipid class composition of the microalga Pavlova lutheri: eicosapentaenoic and docosahexaenoic acids

The lipid classes of Pavlova lutheri, cultivated in semicontinuous mode, were studied by thin-layer chromatography and gas chromatography in attempts to describe the distribution of fatty acid residues within its lipid pool, with special emphasis on eicosapentaenoic (C20:5n-3, EPA) and docosahexaenoic (C22:6n-3, DHA) acids. Neutral lipids and glycolipids were the major constituents and accounted for approximately 57 and 24% of the total fatty acid residues (TFA), respectively. Phospholipids accounted for approximately 10% of TFA. Two lipid classes, acylated steryl glycosides (SG) and diphosphatidylglycerols (DPG), were eventually identified in P. lutheri for the first time. The nonpolar fraction was mainly composed of triacylglycerol (TAG), whereas the polar fraction was mainly composed of monogalactosylacylglycerols (MGDG). The distribution of total EPA and DHA within the lipid pool was calculated in attempts to ascertain the quality of said microalgae as a feed source, as well as the possibility of enhancement of individual fatty acid production and extraction thereafter. EPA was especially concentrated in MGDG (approximately 45%) and TAG (approximately 33%); conversely, DHA was dispersed through various classes, especially within TAG (approximately 27%), DPG (approximately 22%), and betaine lipids (21%).     

Synthesis of structured lipids containing medium-chain and omega-3 fatty acids

The ability of different lipases to incorporate omega3 fatty acids, namely, eicosapentaenoic acid (EPA, C20:5n-3), docosapentaenoic acid (DPA, C22:5n-3), and docosahexaenoic acid (DHA, C22:6n-3), into a high-laurate canola oil, known as Laurical 35, was studied. Lipases from Mucor miehei (Lipozyme-IM), Pseudomonas sp. (PS-30), and Candida rugosa (AY-30) catalyzed optimum incorporation of EPA, DPA, and DHA into Laurical 35, respectively. Other lipases used were Candida anatrctica (Novozyme-435) and Aspergillus niger (AP-12). Response surface methodology (RSM) was used to obtain a maximum incorporation of EPA, DPA, and DHA into high-laurate canola oil. The process variables studied were the amount of enzyme (2-6%), reaction temperature (35-55 degrees C), and incubation time (12-36 h). The amount of water added and mole ratio of substrates (oil to n-3 fatty acids) were kept at 2% and 1:3, respectively. The maximum incorporation of EPA (62.2%) into Laurical 35 was predicted at 4.36% of enzyme load and 43.2 degrees C over 23.9 h. Under optimum conditions (5.41% enzyme; 38.7 degrees C; 33.5 h), the incorporation of DPA into high-laurate canola oil was 50.8%. The corresponding maximum incorporation of DHA (34.1%) into Laurical 35 was obtained using 5.25% enzyme, at 43.7 degrees C, over 44.7 h. Thus, the number of double bonds and the chain length of fatty acids had a marked effect on the incorporation omega3 fatty acids into Laurical 35. EPA and DHA were mainly esterified to the sn-1,3 positions of the modified oils, whereas DPA was randomly distributed over the three positions of the triacylglycerol molecules. Meanwhile, lauric acid remained esterified mainly to the sn-1 and sn-3 positions of the modified oils. Enzymatically modified Laurical 35 with EPA, DPA, or DHA had higher conjugated diene (CD) and thiobarbituric acid reactive substance (TBARS) values than their unmodified counterpart. Thus, enzymatically modified oils were more susceptible to oxidation than their unmodified counterparts, when both CD and TBARS values were considered.     

Effect of aqueous and lipophilic mullet (Mugil cephalus) Bottarga extracts on the growth and lipid profile of intestinal Caco-2 cells

The importance of n-3 polyunsaturated fatty acid (n-3 PUFA) intake has long been recognized in human nutrition. Although health benefits, n-3 PUFA are subject to rapid and/or extensive oxidation during processing and storage, resulting in potential alteration in nutritional composition and quality of food. Bottarga, a salted and semi-dried mullet ( Mugil cephalus ) ovary product, is proposed as an important source of n-3 PUFA, having high levels of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). In this work, we investigated the extent of lipid oxidation of grated bottarga samples during 7 months of storage at -20 °C and room temperature under light exposure. Cell viability, lipid composition, and lipid peroxidation were measured in intestinal differentiated Caco-2 cell monolayers after 6-48 h of incubation with lipid and hydrophilic extracts obtained from bottarga samples at different storage conditions. The storage of bottarga did not affect the n-3 PUFA level, but differences were observed in hydroperoxide levels in samples from different storage conditions. All tested bottarga extracts did not show a toxic effect on cell viability of differentiated Caco-2 cells. Epithelial cells incubated with bottarga oil had significant changes in fatty acid composition but not in cholesterol levels with an accumulation of EPA, DHA, and 22:5. Cell hydroperoxides were higher in treated cells, in relation to the oxidative status of bottarga oil. Moreover, the bottarga lipid extract showed an in vitro inhibitory effect on the growth of a colon cancer cell line (undifferentiated Caco-2 cells).     

Lipid characterization of mangrove thraustochytrid--Schizochytrium mangrovei

Lipid class composition and distribution of fatty acids within the lipid pool of microalga, Schizochytrium mangrovei FB3 harvested at the late exponential phase, was studied, with special emphasis on the distribution of docosahexaenoic acid (C22:6 n-3, DHA). Neutral lipids were the major lipid constituents (95.90% of total lipids) in which triacylglyerol (TAG) was the predominant component and accounted for 97.20% of the neutral lipids. Phosphatidylcholine (PC) was the major polar lipid. Phosphatic acid and phosphatidylserine were the two classes in phospholipids reported for the first time in thraustochytrids. Both TAG and PC were primarily saturated and consisted of C16:0 at approximately 50% of total fatty acids. DHA was found to be distributed in all lipid classes and to be the major polyunsaturated fatty acid. TAG contained the highest amount of DHA, although the percentage of DHA in total fatty acids in TAG (29.74%) was lower than that in PC (39.61%). The result from this study would be useful for further optimization of DHA production by S. mangrovei.     

Effects of season and processing on oil content and fatty acids of baltic herring (Clupea harengus membras)

Fatty acid composition, oil content, free fatty acid content, and peroxide value of Baltic herring (Clupea harengus membras) and two processed products (fried fillets and fish burgers) were investigated. The highest oil content of the fillets was found in autumn (10%), at the time when the free fatty acids had their minimum (1.4%). The main fatty acids were oleic (18-23%), palmitic (17%), palmitoleic (8-12%), and docosahexaeneoic (8-10%) acids. The proportion of saturated fatty acids was a constant 23% all year around, whereas mono- and polyunsaturated acids varied from 34 to 39% and 33 to 37%, respectively. During processing the oil content doubled and the fatty acid composition changed to the pattern of the rapeseed oil used for frying. Oleic acid was a major fatty acid in the products comprising over 40% of the total fatty acids. The proportion of n-3 acids decreased during processing but the total amount of polyunsaturated acids remained fairly constant.     

Baking reduces prostaglandin, resolvin, and hydroxy-fatty acid content of farm-raised Atlantic salmon (Salmo salar)

The consumption of seafood enriched in n-3 polyunsaturated fatty acids (PUFA) is associated with a decreased risk of cardiovascular disease. Several n-3 oxidation products from eicosapentaenoic acid (EPA; 20:5n-3) and docosahexaenoic acid (22:6n-3) have known protective effects in the vasculature. It is not known whether the consumption of cooked seafood enriched in n-3 PUFA causes appreciable consumption of lipid oxidation products. We tested the hypothesis that baking Atlantic salmon (Salmo salar) increases the level of n-3 and n-6 PUFA oxidation products over raw salmon. We measured the contents of several monohydroxy-fatty acids (MHFA), prostanoids, and resolvins. Our data demonstrate that baking did not change the overall total levels of MHFA. However, baking resulted in selective regioisomeric loss of hydroxy fatty acids from arachidonic acid (20:4n-6) and EPA, while significantly increasing hydroxyl-linoleic acid levels. The contents of prostanoids and resolvins were reduced several-fold with baking. The inclusion of a coating on the salmon prior to baking reduced the loss of some MHFA but had no effect on prostanoid losses incurred by baking. Baking did not decrease n-3 PUFA contents, indicating that baking of salmon is an acceptable means of preparation that does not alter the potential health benefits of high n-3 seafood consumption. The extent to which the levels of MHFA, prostanoids, and resolvins in the raw or baked fish have physiologic consequence for humans needs to be determined.     

Fatty acid composition and squalene content of the marine microalga Schizochytrium mangrovei

Certain species of thraustochytrids are being explored as potential producer of polyunsaturated fatty acids for nutritional enrichment of food products and use as feed additives in aquaculture. The fatty acid composition and squalene content were determined in the thraustochytrid, Schizochytrium mangrovei that was newly isolated from decaying Kandelia candel leaves in Hong Kong mangrove habitat. The major fatty acid constituents identified in all three S. mangrovei strains were tetradeanoic acid (C14:0), hexadecanoic acid (C16:0), docosapentaenoic acid (C22:5 n-6, DPA) and docosahexaenoic acid (C22:6 n-3, DHA). DHA was the most predominant polyunsaturated fatty acid, and the percentage of DHA (of total fatty acids) in all these strains varied from 32.29 to 39.14%. Only slight changes were observed in fatty acid composition of the S. mangrovei strains harvested at their early (day 3) and late stationary (day 5) phases. In contrast, the cellular squalene content was affected significantly by the culture time; the largest decrease of squalene content from 0.162 mg/g to 0.035 mg/g was found in S. mangrovei FB1 as the culture aged.     

Free polyunsaturated fatty acids cause taste deterioration of salmon during frozen storage

Increased intensity of train oil taste, bitterness, and metal taste are the most pronounced sensory changes during frozen storage of salmon (Refsgaard, H. H. F.; Brockhoff, P. B.; Jensen, B. Sensory and Chemical Changes in Farmed Atlantic Salmon (Salmo salar) during Frozen Storage. J. Agric. Food Chem. 1998a, 46, 3473-3479). Addition of each of the unsaturated fatty acids: palmitoleic acid (16:1, n - 7), linoleic acid (C18:2, n - 6), eicosapentaenoic acid (EPA; C20:5, n - 3) and docosahexaenoic acid (DHA; C22:6, n - 3) to fresh minced salmon changed the sensory perception and increased the intensity of train oil taste, bitterness, and metal taste. The added level of each fatty acid ( approximately 1 mg/g salmon meat) was equivalent to the concentration of the fatty acids determined in salmon stored as fillet at -10 degrees C for 6 months. The effect of addition of the fatty acids on the intensity of train oil taste, bitterness and metal taste was in the order: DHA > palmitoleic acid > linoleic acid > EPA. Formation of free fatty acids was inhibited by cooking the salmon meat before storage. Furthermore, no changes in phospholipid level were observed during frozen storage. The results suggest that enzymatic hydrolysis of neutral lipids plays a major role in the sensory deterioration of salmon during frozen storage.     

Effect of the polyunsaturated fatty acid composition of beef muscle on the profile of aroma volatiles

The effect of n-3 polyunsaturated fatty acids (PUFAs) in beef muscle on the composition of the aroma volatiles produced during cooking was measured. The meat was obtained from groups of steers fed different supplementary fats: (i) a palm-oil-based control; (ii) bruised whole linseed, which increased muscle levels of alpha-linolenic (C18:3 n-3) and eicosapentaenoic acid (EPA, C20:5 n-3); (iii) fish oil, which increased EPA and docosahexaenoic acid (C22:6 n-3); (iv) equal quantities of linseed and fish oil. Higher levels of lipid oxidation products were found in the aroma extracts of all of the steaks with increased PUFA content, after cooking. In particular, n-alkanals, 2-alkenals, 1-alkanols, and alkylfurans were increased up to 4-fold. Most of these compounds were derived from the autoxidation of the more abundant mono- and di-unsaturated fatty acids during cooking, and such autoxidation appeared to be promoted by increased levels of PUFAs.     

Variation in fatty acid composition of Artemia salina nauplii enriched with microalgae and baker's yeast for use in larviculture

The high content of the essential fatty acids in some microalgae and baker's yeast has made them excellent diets for boosting the fatty acid content of livefood Artemia. The influences of baker's yeast (Saccharomyces cerevisiae) and three microalgae, viz., Chlorella salina, Chaetoceros calcitrans, and Nannochloropsis salina, were tested as diet components in marine livefeed brine shrimp Artemia salina nauplii to improve the polyunsaturated fatty acid (PUFA) composition. Artemia nauplii submerged in these diets for four different enrichment intervals (3, 6, 8, and 24 h) were found to incorporate essential fatty acids, and the percentage composition of different fatty acids was measured in the enriched Artemia nauplii and enrichment diets. N. salina produced higher levels of arachidonic acid (AA, 20:4n6, 9.50%), eicosapentaenoic acid (EPA, 20:5n3, 25.80%), and docosahexaenoic acid (DHA, 22:6n3, 4.18%) as compared to other diets. The total PUFA content of the enriched Artemia by N. salina increased by 56.50% with enrichment periods up to 8 h, followed by a significant reduction in the final 24 h. N. salina yielded Artemia nauplii with considerable EPA (8.05%), AA (14.15%), and DHA (1.85%) after 8 h of enrichment, which are significantly higher levels than in nauplii fed with the other three diets (p = 0.05). The DHA/EPA values in Artemia enriched for 6 h by N. salina and C. calcitrans were found to be, respectively, 88.46 and 25% higher than freshly hatched Artemia. Artemia enriched by C. salina and baker's yeast exhibited a reduction in PUFA content even at 6 h of enrichment. Significant relative decreases in DHA, EPA, and total PUFA in Artemia enriched with all of the diets were apparent, with a corresponding increase in the total saturated fatty acid content (26.95 +/- 9.75%) in the final stages (24 h) of enrichment (p = 0.05).     

Compositional characteristics and antioxidant properties of fresh and processed sea cucumber (Cucumaria frondosa)

The antioxidant activity of fresh and rehydrated sea cucumber (Cucumaria frondosa) samples with/without internal organs was evaluated for the first time. In addition, their proximate, amino acid, and fatty acid compositions were examined. Rehydrated sea cucumber samples in distilled water were prepared from oven-dried products. All samples contained 83-90% moisture, but showed a significant difference among groups in their protein and lipid contents. Glutamic acid was the predominant amino acid in sea cucumber, followed by glycine and aspartic acid. Essential amino acids such as leucine and lysine were also present at high levels. The trend for free amino acid was different from that of total amino acids and varied among groups. Lipids in sea cucumber were dominated by eicosapentaenoic acid (EPA, C20:5n-3), ranging from 43.2 to 56.7% of the total fatty acids. Docosahexaenoic acid (DHA, C22:6n-3) was present at a much lower concentration of 2.0-5.8%. All sea cucumber samples exhibited radical scavenging property against 2,2'-azobis(2-aminopropane) dihydrochloride (AAPH) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals, with rehydrated samples, especially those with internal organs, possessing higher antioxidant activity than their fresh counterparts. No correlation existed between radical scavenging capacity and total phenolics content, suggesting that other components, in addition to phenolic compounds, contribute to the antioxidant activity of sea cucumber.     

Lipid [corrected] classes, fatty acids, and sterols in seafood from Gilbert Bay, southern labrador

Seafood from Gilbert Bay, southern Labrador, was sampled for lipid classes, fatty acid, and sterol composition. Gilbert Bay is a proposed Marine Protected Area, and the composition of seafood from this region is interesting from both human health and ecological perspectives. Analyses included four species of bivalves and flesh and liver samples from four fish species. Lipids from a locally isolated population of northern cod (Gadus morhua) were also compared to lipids from other cod populations. Lipid classes were analyzed by Chromarod/Iatroscan TLC-FID, fatty acids by GC, and sterols by GC-MS. Three cod populations had similar levels of total lipid per wet weight (0.6%) with triacylglycerols (TAG), sterols, and phospholipids comprising on average 13, 11, and 51%, respectively, of their total lipids. Fatty fish such as capelin and herring contained on average 8.4% lipid with 86% present as TAG. Fish livers from cod and herring showed opposite trends, with cod having elevated lipid (27%) and TAG (63%) and herring containing only 3.8% lipid and 20% TAG. Shellfish averaged 0.6% lipid; however, significant lipid class differences existed among species. Fatty acid analysis showed few significant differences in cod populations with on average 57% polyunsaturated fatty acids (PUFA), 18% monounsaturated fatty acids (MUFA), and 24% saturated fatty acids (SFA). Cod livers had lower PUFA (34%) and elevated MUFA (44%) relative to flesh. Bivalves averaged 25% SFA, 18% MUFA, and 57% PUFA, whereas scallop adductor muscle had the highest PUFA levels (63%). Bivalves contained 20 different sterols with cholesterol present as the major sterol (19-39%). trans-22-Dehydrocholesterol, brassicasterol, 24-methylenecholesterol, and campesterol individually accounted for >10% in at least one species. High levels of PUFA and non-cholesterol sterols observed in Gilbert Bay seafood demonstrate their positive attributes for human nutrition.     

富含EPA/DHA型结构磷脂的酶法合成条件优化及表征

为建立富含二十碳五烯酸(EPA)和二十二碳六烯酸(DHA)型结构磷脂的制备方法,本试验以南极磷虾磷脂为原料,以EPA和DHA总结合率为指标,通过响应面法优化富含EPA/DHA型结构磷脂的酶法合成反应参数,并采用气相色谱(GC)和亲水作用色谱-质谱联用(HILIC-MS)技术对其结构进行表征。结果表明,最优酶催化反应条件为:底物质量比(游离脂肪酸:磷脂, m/m)5.15,反应温度55.22℃,水分添加量(以反应底物总质量计)0.92%,酶添加量(以反应底物总质量计)20%,正己烷3 mL,反应时间24 h,所得结构磷脂中EPA/DHA结合率达64.35%;经GC和HILIC-MS综合分析可知,结构磷脂中磷脂酰胆碱(PC)亚类40:10(20:5/20:5)(18.46%),42:11(20:5/22:6)(12.78%)和38:6(18:1/20:5&16:0/22:6) (7.07%)等分子种含量较高;溶血磷脂酰胆碱LPC亚类20:5(20:5/0:0)(34.40%),22:6(22:6/0:0)(25.99%)和O-16:0(O-16:0/0:0)(10.22%)等分子种含量较高;结构磷脂的PC和LPC组分中脂肪酸单链平均长度≥20的分子种相对含量较原料磷脂分别提高了33.12和37.28个百分点;结构磷脂的PC组分中双键数≥5和双键数≥10的分子种相对含量较原料磷脂分别提高了24.89和27.53个百分点,表明该方法有效提高了结构磷脂中EPA和DHA的含量。本研究结果为进一步探索和丰富酶促合成EPA/DHA型结构磷脂的机理和实践提供了一定的数据支持。     

不同产地海带脂肪酸组成的比较分析

为比较分析不同产地海带的脂肪酸组成,本研究采用溶剂法对海带总脂提取条件进行优化,并通过气相色谱-质谱技术对福州、宁波和青岛3个产地海带的总脂脂肪酸组成进行比较分析。结果表明,二氯甲烷-甲醇法提取海带总脂的效率较高,是海带总脂提取的理想方法;福州、宁波和青岛海带的总脂含量依次为0.75%、0.69%和0.86%,且无显著差异。从海带总脂中共鉴定出30种脂肪酸,以 C16:0、C18:1n-9、C20:4n-6(AA)和C20:5n-3(EPA)为主,且3个产地海带在脂肪酸组成上存在显著差异。福州产地海带中的C20:4n-6和C20:5n-3含量分别为10.12%、6.18%,其中C20:4n-6含量显著高于宁波(9.41%)和青岛(8.77%),C20:5n-3含量显著低于宁波(8.13%)和青岛(9.40%),进而使3个产地海带中的n-3 PUFA、n-6 PUFA、n-3/n-6、EPA+AA和EPA/AA等指标存在显著差异。本研究结果为不同产地海带总脂提取、脂肪酸比较分析及营养评价提供了一定的理论依据。     

Improving the fatty acid profile of fairy shrimp, Streptocephalus dichotomus, using a lipid emulsion rich in highly unsaturated fatty acids

Fatty acids are the largest component of lipids and have become a useful tool in the determination of live feeds to a variety of cultured species. Bioencapsulation is a technique which allows high-level incorporation of desired components (i.e., fatty acids, vitamins, antibiotics, etc.) in live feeds, which in turn can be supplemented to the consumer organisms. The procedure described in the present study serves as a platform of technology for enriching the Streptocephalus dichotomus. Uptake of two enrichment diets (ALGAMAC2000 and DHA-SELCO) by adult S. dichotomus was investigated. The fatty acid profile supports the hypothesis that the enrichment diet increases the level of essential fatty acids, such as linolic, linolenic, eicosapentenoic, and docosahexaenoic acids. The average content (percent of total fatty acids detected) of the enriched organism by different highly unsaturated fatty acid (HUFA) products were as follows: ALGAMAC2000 showed 14-22% saturated fatty acid (SFA), 17-18% monounsaturated fatty acid (MUFA), 28-41% polyunsaturated fatty acid (PUFA), 23-34% n-3, and 4.9-7.5% n-6, whereas DHA-SELCO showed about 20-23% SFA, 20-26% MUFA, 38% PUFA, 28-31% n-3, and 7.5-10% n-6. Our present investigation proves that both HUFA-rich diets appear to be an appropriate enrichment diet, and further provides an additional rationale for using fairy shrimp as a maturation diet for any cultivable freshwater organism.     

Up-regulated desaturase and elongase gene expression promoted accumulation of polyunsaturated fatty acid (PUFA) but not long-chain PUFA in Lates calcarifer, a tropical euryhaline fish, fed a stearidonic acid- and γ-linoleic acid-enriched diet

The limited activity of Δ6 fatty acid desaturase (FAD6) on α-linolenic (ALA, 18:3n-3) and linoleic (LA, 18:2n-6) acids in marine fish alters the long-chain (≥C(20)) polyunsaturated fatty acid (LC-PUFA) concentration in fish muscle and liver when vegetable oils replace fish oil (FO) in aquafeeds. Echium oil (EO), rich in stearidonic acid (SDA, 18:4n-3) and γ-linoleic acid (GLA, 18:3n-6), may enhance the biosynthesis of n-3 and n-6 LC-PUFA by bypassing the rate-limiting FAD6 step. Nutritional and environmental modulation of the mechanisms in LC-PUFA biosynthesis was examined in barramundi, Lates calcarifer , a tropical euryhaline fish. Juveniles were maintained in either freshwater or seawater and fed different dietary LC-PUFA precursors present in EO or rapeseed oil (RO) and compared with FO. After 8 weeks, growth of fish fed EO was slower compared to the FO and RO treatments. Irrespective of salinity, expression of the FAD6 and elongase was up-regulated in fish fed EO and RO diets, but did not lead to significant accumulation of LC-PUFA in the neutral lipid of fish tissues as occurred in the FO treatment. However, significant concentrations of eicosapentaenoic acid (EPA, 20:5n-3) and arachidonic acid (ARA, 20:4n-6), but not docosahexaenoic acid (DHA, 22:6n-3), appeared in liver and, to a lesser extent, in muscle of fish fed EO with marked increases in the phospholipid fraction. Fish in the EO treatment had higher EPA and ARA in their liver phospholipids than fish fed FO. Endogenous conversion of dietary precursors into neutral lipid LC-PUFA appears to be limited by factors other than the initial rate-limiting step. In contrast, phospholipid LC-PUFA had higher biosynthesis, or selective retention, in barramundi fed EO rather than RO.     

Enzyme-assisted acidolysis of borage (Borago officinalis L.) and evening primrose (Oenothera biennis L.) oils: incorporation of omega-3 polyunsaturated fatty acids.

Lipase-catalyzed acidolysis of borage (Borago officinalis L.) and evening primrose (Oenothera biennisL.) oils with long-chain omega3 polyunsaturated fatty acids (PUFA), namely, eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids, was carried out in hexane, and the products were analyzed using gas chromatography. The most effective lipase for incorporation of omega3 PUFA into these oils was Pseudomonas sp. as compared to lipases from Mucor miehei and Candida antarctica. Response surface methodology was used to obtain a maximum yield of EPA+DHA incorporation while using the minimum amount of enzyme possible. The process variables studied were the amount of enzyme (150-350 units), reaction temperature (30-60 degrees C), and reaction time (6-30 h). All experiments were carried out according to a face-centered cube design. Under optimum conditions, incorporation of EPA+DHA was 35.5% in borage oil and 33. 6% in evening primrose oil. The modified borage and evening primrose oils containing gamma-linolenic acid, EPA, and DHA were successfully produced and may have potential health benefits.     

Enrichment of eicosapentaenoic acid from sardine oil with Delta5-olefinic bond specific lipase from Bacillus licheniformis MTCC 6824

Lipase derived from Bacillus licheniformis MTCC 6824 was purified to homogeneity by anion exchange chromatography on Amberlite IRA 410 (Cl-) and gel filtration using Sephadex G-100 as judged by denaturing polyacrylamide gel electrophoresis. The purified lipase was used for hydrolysis of triacylglycerol in sardine oil to enrich Delta5-polyunsaturated fatty acids (Delta5-PUFAs) namely, arachidonic acid (5,8,11,14-eicosatetraenoic acid, ARA, 20:4n-6) and eicosapentaenoic acid (5,8,11,14,17-eicosapentaenoic acid, EPA, 20:5n-3). The individual fatty acids were determined as fatty acid methyl esters (FAMEs) by gas-liquid chromatography and gas chromatography-mass spectroscopy as FAMEs and N-acyl pyrrolidides. The enzyme exhibited hydrolytic resistance toward ester bonds of Delta5-PUFAs as compared to those of other fatty acids and was proved to be effective for increasing the concentration of EPA and ARA from sardine oil. Utilizing this fatty acid specificity, EPA and ARA from sardine oil were enriched by lipase-mediated hydrolysis followed by urea fractionation at 4 degrees C. The purified lipase produced the highest degree of hydrolysis for SFAs and MUFAs (81.5 and 72.3%, respectively, from their initial content in sardine oil) after 9 h. The profile of conversion by lipase catalysis showed a steady increase up to 6 h and thereafter plateaued down. Lipase-catalyzed hydrolysis of sardine oil followed by urea adduction with methanol provided free fatty acids containing 55.4% EPA and 5.8% ARA, respectively, after complexation of saturated and less unsaturated fatty acids. The combination of enzymatic hydrolysis and urea complexation proved to be a promising method to obtain highly concentrated EPA and ARA from sardine oil.     

Annual evolution of fatty acid profile from muscle lipids of the common carp (Cyprinus carpio) in Madagascar inland waters

Annual evolution of muscle lipids fatty acid (FA) from common carp (Cyprinus carpio) has been determined in 2001 through monthly samplings in the reserve pond of Sisaony (SIS series) and Itasy Lake (ITA series) of the Madagascar highlands. Total lipids from muscle were extracted and quantified according to the Bligh and Dyer method. FA identification was performed by GC-MS of FA methyl esters and FA pyrrolidides and led to the identification of 41 FA; routine analyses of FA were made by capillary GC. Principal component analysis (PCA) was performed on the data set to compare FA profiles. Lipid content is low, ranging from 0.91 to 1.73% of wet muscle, with a low stage during the hot season (January-April) and a higher stage during the cold season (July-October). Three FA dominated the FA composition: oleic acid (17.0-21.5%), palmitic acid (13.1-16.1%), and linoleic acid (9.6-13.2%). Polyunsaturated fatty acids (PUFA) were present in appreciable amounts: arachidonic acid (AA; 2.9-5.9%), docosahexaenoic acid (DHA; 2.9-6.7%), eicosapentaenoic acid (EPA; 1.9-3.4%), and docosapentaenoic acid (DPA; 1.9-4.3%). Two opposite evolution schemes appear within two groups of FA; on the one hand PUFA (both n-3 and n-6 series) show a maximum in August-October and a minimum in January-April, and, on the other hand, oleic, palmitic, and linoleic acids show the opposite maxima and minima. PCA results give confirmation of these evolution schemes, the two groups of FA giving opposite high factor loadings on axis 1. The SIS and ITA series are differentiated by axis 2 by mean of minor FA, mostly odd- and branched-chain. Results indicate that common carp, the second most abundant freshwater fish in Madagascar highlands waters, may be an interesting source of dietary PUFA.