n-3多不饱和脂肪酸富集纯化的研究进展

n-3多不饱和脂肪酸为一类独特的生物活性物质,在生物体内具有广泛的生理功能.随着对其生理功能的认识和研究的不断深入,对n-3多不饱和脂肪酸富集纯化的研究也成为热点.主要介绍n-3多不饱和脂肪酸的富集纯化方法,并对植物种子中α-亚麻酸的富集以及鱼油中DHA和EPA富集的研究进行了综述.     

脂肪酶催化醇解制备富含n-3 PUFAs的甘油酯

n-3 PUFAs具有多种生理活性。以鱼油为原料,采用脂肪酶催化醇解法对其中的n-3 PUFAs进行富集,得到富含n-3 PUFAs的甘油酯。结果表明：来自米曲霉的脂肪酶Eversa Transform 2.0(ET 2.0)对n-3 PUFAs表现出良好的醇解特异性。通过单因素实验确定最佳工艺条件为：甲醇体积分数40%,m甲醇∶m鱼油=1.6∶1,温度35℃,脂肪酶添加量4%,反应时间2 h。在此条件下,鱼油甘油酯中n-3 PUFAs的含量从39.88%上升至67.87%,EPA和DHA的含量从36.61%上升至65.22%,n-3 PUFAs的得率为56.33%。脂肪酶在最佳反应条件下对不同底物的富集效果是：金枪鱼油甘油酯中n-3 PUFAs的含量从38.81%上升到61.35%,藻油甘油酯中n-3 PUFAs的含量从45.96%上升到64.09%。上述方法获得的富含n-3PUFAs甘油酯在食品和制药行业有很大的应用潜力。     

功能性n-3多不饱和脂肪酸的研究进展

脂肪酸是生命代谢活动中一种重要化合物,根据脂肪酸碳链数上的双键数将脂肪酸分为饱和脂肪酸、单不饱和脂肪酸、多不饱和脂肪酸三类。文章对n-3多不饱和脂肪酸来源、合成过程、n-3和n-6多不饱和脂肪酸的结构差异以及生物活性进行综述,为n-3多不饱和脂肪酸在功能保健食品、医学药用领域及基础研究进一步探索与全面开发提供理论依据。     

鱼油n-3 PUFA富集、稳态化技术及生物活性研究进展

鱼油富含人体所需的多不饱和脂肪酸,在科研与应用领域具有广泛价值。本文重点围绕其n-3 PUFA富集工艺、稳态化技术及生物活性进行综述,阐述不同方法的优、缺点,分析工业化生产的可行性,展望未来鱼油加工领域的发展趋势,旨在为鱼油类资源的合理、健康、可持续发展提供理论依据。     

超声辅助脂肪酶催化合成富含n-3多不饱和脂肪酸磷脂的研究

以脂肪酶Lipozyme TL IM为催化剂,研究了合成富含n-3多不饱和脂肪酸磷脂的最佳工艺条件,即以正庚烷:异辛烷:正已烷=8:1:1为超声介质,溶剂(mL)/底物(g)=2,酶用量为25%,超声介质中水分质量分数为2%,超声功率70 W,55℃下连续超声5.5 h,在此条件下酯交换到磷脂上的DHA和EPA总质量分数为22.99%。由此可见,通过超声辅助及酯交换条件的优化,不仅可以大大缩短反应所需的时间,而且也能充分激活Lipozyme TL IM的酶活力,从而提高酯化反应的效率。     

高n-3多不饱和脂肪酸鸡蛋的开发研究进展

介绍了n-3多不饱和脂肪酸类别、营养功能及使用哪些饲料原料可以生产高n-3多不饱和脂肪酸鸡蛋,分析了鸡蛋中多不饱和脂肪酸的沉积规律和影响因素,并概述了高n-3多不饱和脂肪酸鸡蛋特点,以及存在的问题和解决途径,为开发高n-3多不饱和脂肪酸鸡蛋提供依据。     

有机溶剂中酶促酯化合成n-3 PUFA甘油酯的研究

利用脂肪酶催化游离型的n-3PUFA与甘油发生酯化反应生成甘油酯,研究了多种因素对酯化程度的影响.研究结果表明：利用脂肪酶Novozym435,在6mL的正己烷中,反应温度40℃,甘油用量2g,n-3PUFA0.4g,初始水分含量0.5%,在反应1h后添加1g分子筛,酶添加量25mg,震荡频率150r/min,在此条件下反应24h酯化度可达90%以上.n-3 PUFA甘油酯中以甘油二酯（56.06%）为主,其次是甘油三酯（31.25%）和甘油一酯（12.07%）,游离脂肪酸（0.39%）的含量比较低.n-3 PUFA甘油酯的脂肪酸组成以DHA（73.36%）和EPA（13.49%）为主,单不饱和脂肪酸的含量仅有1.69%,几乎不合饱和脂肪酸.     

n-3多不饱和脂肪酸分类、来源与疾病防治功能

n-3多不饱和脂肪酸包括ALA、EPA、DPA和DHA等。增加n-3多不饱和脂肪酸摄取量可以促进婴幼儿视网膜、大脑和神经系统发育；n-3多不饱和脂肪酸通过各种途径降低人体心血管疾病和炎症的发生，降低糖尿病患者血清LDL-C和TG水平，抑制体外培养的乳腺、前列腺和结肠癌细胞增生，促进细胞凋亡。     

酶促鱼油选择性水解制备EPA、DHA甘油酯的研究

研究了假丝酶母脂肪酶催化鱼油选择性水解反应 ,确定了油水比、脂肪酶浓度、反应温度、pH、激活剂、溶剂、反应时间、水解率等因素对反应的影响 ,同时对鱼油水解产物进行了分离和检测 ,使EPA、DHA在甘油酯型产品中含量达到 5 0 %以上     

酶法制备甘油二酯的研究进展北大核心CSCD

甘油二酯(DAG)可抑制体内脂肪堆积、降低血清甘油三酯水平从而具有减肥效果。与化学法相比,酶法制备DAG具有反应条件温和、选择性强、环境友好等优点。综述了国内外酶法制备DAG的研究现状,对酶促酯化合成、酶促甘油解以及酶促水解法制备DAG进行了详细介绍,着重阐述了如何提高DAG的选择性,总结了脂肪酶在DAG制备中的关键问题,以期为DAG的工业化生产提供参考。     

Production of n-3 polyunsaturated fatty acid concentrate from sardine oil by immobilized Candida rugosa lipase

ABSTRACT:  This study was conducted to develop an immobilized-enzyme system to entrap lipase in chitosan-alginate-CaCl₂ beads for the purpose of concentrating n-3 polyunsaturated fatty acids (n-3 PUFAs) from sardine oil. Lipase was immobilized by an ionotropic gelatin method analyzed for characteristics. Optimum pH of immobilized lipase shifted from pH 7.0 to 6.0 and immobilized lipase showed higher stability against pH and temperature changes. Original sardine oil contained 38.1% n-3 PUFAs (25.2% 20:5n3 and 7.20% 22:6n3), and the concentration was significantly increased to 65.3% (40.2% 20:5n3 and 15.5% 22:6n3) with free lipase and to 64.8% (39.6% 20:5n3 and 15.3% 22:6n3) with immobilized lipase after 90 min of repeated hydrolysis. Fatty acid content of the free fatty acid (FFA) fraction of hydrolyzed oil showed that lipase preferably hydrolyzed 16:0, 16:1n7 and 18:0 accounting for 76.6% and 69.5% of total FFAs (after 1st and 2nd hydrolysis, respectively). This study shows that use of immobilized lipase systems for increasing n-3 PUFA concentration in sardine oil provides new processing opportunities for the industry.     

n-3 polyunsaturated fish fatty acids in a fish-oil-supplemented bread

White bread enriched in n-3 fatty acids in the form of gelatine-coated fish oil and marketed in Denmark since 1990 under the name of ‘Omega Bread’, was found to be a reliable and significant source of higher n-3 polyunsaturated fatty acids (with 20 and 22 C atoms). The bread from six bakeries contained 40-55 mg per 100 g more n-3 fatty acids. By eating 200 g daily of this bread one gets some 25-30% of the amount of higher n-3 polyunsaturated fatty acids present in 30 g of mixed fish. The daily ingestion of this amount of fish over a period of 20 years is known to reduce mortality from coronary heart disease by more than 50%. The supplement of n-3 polyunsaturated fatty acids obtained by eating Omega Bread daily in place of other types of white bread is expected to reduce coronary heart disease. In addition, such an increased daily ingestion of n-3 polyunsaturated fatty acids over a long-term period may possibly have other beneficial health effects. It is recommended that the intake of higher n-3 fatty acids in industrialised Western societies should be increased by more than can be obtained by a daily ration of Omega Bread. It is therefore advisable that consumption of Omega Bread is combined with an increased intake of fish and/or fish oil capsules and/or fish-oil-enriched substitutes for butter and margarine.     

Effects of feeding rumen-protected linseed fat to postpartum dairy cows on plasma n-3 polyunsaturated fatty acid concentrations and metabolic and reproductive parameters

High-yielding dairy cows experience a negative energy balance and inflammatory status during the transition period. Fat supplementation increases diet energy density, and plasma n-3 polyunsaturated fatty acids (PUFA) have been proposed to improve immune function. This study tested the hypothesis that dietary supplementation with a rumen-protected and n-3 PUFA-enriched fat could ameliorate both the energetic deficit and immune status of postpartum high-yielding dairy cows, improving overall health and reproductive efficiency. At 11 d in milk (DIM), cows were randomly allocated to groups (1) n-3 PUFA (n = 29), supplemented with encapsulated linseed oil supplying additional up to 64 g/d (mean 25 ± 4 g/d) of α-linolenic acid (ALA), or (2) control (n = 31), supplemented with hydrogenated palm oil without ALA content. Fat supplements of the n-3 PUFA and control groups were available through an automated, off-parlor feeding system, and intake depended on the cow's feeding behavior. Plasma ALA concentrations were higher in n-3 PUFA than control cows, following a linear relation with supplement ingestion, resulting in a lower n-6/n-3 ratio in plasma. Metabolic parameters (body condition score and glucose and β-hydroxybutyric acid blood concentrations) were unaffected, but milk yield improved with increased intake of fat supplements. Plasma total adiponectin concentrations were negatively correlated with ingestion of n-3 PUFA-enriched fat supplement, following a linear relation with intake. Conception rate to first AI increased with higher intake of both fats, but a decrease of calving-to-conception interval occurred only in n-3 PUFA cows. Postpartum ovarian activity and endometrial inflammatory status at 45 DIM were unaffected. In conclusion, this study evinced a positive linear relation between rumen-protected linseed fat intake and plasma n-3 PUFA concentrations, which modulated adiponectin expression and improved reproductive parameters.     

n-3多不饱和脂肪酸对疾病的预防与治疗作用

n-3多不饱和脂肪酸主要源于亚麻籽、胡桃仁、大豆以及海洋生物。增加n-3多不饱和脂肪酸摄取量可以促进视网膜、大脑和神经系统发育。n-3多不饱和脂肪酸通过各种途径降低人体心血管疾病和炎症的发生,降低糖尿病患者血清低密度脂蛋白胆固醇(LDL-C)和甘油三酯(TG)水平。膳食中增加n-3多不饱和脂肪酸的含量可以对心血管疾病、糖尿病、炎症等患者有辅助治疗效果;n-3多不饱和脂肪酸可以作用于神经细胞,对注意力不集中、抑郁症、精神分裂症有一定的疗效。n-3多不饱和脂肪酸可抑制体外培养的乳腺、前列腺和结肠癌细胞增生,促进细胞凋亡。     

Repeated hydrolysis process is effective for enrichment of omega 3 polyunsaturated fatty acids in salmon oil by Candida rugosa lipase

Enrichment of omega 3 polyunsaturated fatty acids (PUFA) in the glyceride fraction of salmon oil was performed by Candida rugosa lipase (CRL)-catalysed hydrolysis. Total omega 3 PUFA content in the product was 38.71% (mol.%), more than double of the initial level. The hydrolysis reaction was scaled up to 1 l in a stirred tank reactor without any decrease in the selectivity of the reaction. Moreover, non-steroselectivity and acyl chain specificity of CRL-catalysed hydrolysis, as well as CRL’s recognition of the whole triacylglycerol molecule, were proved. The product was purified by short path distillation, which was also shown to contribute to the recovery of omega 3 PUFA in residue. After the removal of free fatty acids, the final product was subjected to a second round of hydrolysis to concentrate omega 3 PUFA further to 50.58%. Recoveries of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) decreased slightly after the second round of hydrolysis, while oleic acid (OA) content was not affected.     

Plant polyphenols associated with vitamin E can reduce plasma lipoperoxidation in dairy cows given n-3 polyunsaturated fatty acids

Diets rich in n-3 polyunsaturated fatty acids (PUFA) improve the nutritional value of ruminant products but also increase the risk of lipoperoxidation in plasma and tissues. The relative effectiveness of dietary antioxidants such as vitamin E (vit E) given alone or with plant extracts rich in polyphenols (PERP) containing rosemary, grape, citrus, and marigold was investigated in the plasma of mid-lactation dairy cows given diets enriched in 18:3 n-3. For a 30-d period, the animals were given a maize silage-based diet (control group C, n = 6) or the same basal diet supplemented with extruded linseed rich in 18:3 n-3 [50 g of oil/kg of diet dry matter (DM); group L, n = 6], extruded linseed + vit E (375 international units/kg of diet DM; 7,500 IU/cow per day; group LE, n = 6), or extruded linseed + vit E + PERP (10 g/kg of diet DM; group LEP, n = 5). Plasma susceptibility to lipoperoxidation was evaluated using in vitro parameters of conjugated diene formation (lag phase and maximum oxidation rate). Plasma indicators of lipoperoxidation and antioxidant status were analyzed in the 4 experimental groups as well as the fatty acid (FA) composition of total plasma lipids. At d 30, group L significantly increased plasma cholesterol esters (+57%) and phospholipids (+35%) compared with group C. It also increased plasma n-3 PUFA (4.7-fold increase) to the detriment of n-6 PUFA (−30%), leading to a higher peroxidizability index (+20%). Plasma in vitro lipoperoxidation was higher in group L (rich in 18:3 n-3) than in group C. Vitamin E alone had no effect on lipoperoxidation, whereas vit E in association with PERP lowered lipoperoxidation by increasing the resistance time against peroxidation (+47%) and by decreasing the oxidation rate (−48%) compared with group L at d 30. Surprisingly, in vivo plasma lipoperoxidation estimated by the plasma level of the major lipoperoxidation product (malondialdehyde) was not significantly increased in group L. This study shows, for the first time, that PERP supplied in association with vit E were able to reduce lipoperoxidation in lactating cows given a diet rich in 18:3 n-3, thereby helping to protect cows against the deleterious consequences of lipoperoxidation and potentially ensuring antioxidant potential for 18:3 n-3–enriched dairy products.     

Enhancement of dairy sheep cheese eating quality with increased n-3 long-chain polyunsaturated fatty acids

This study investigated the effect of different plant oil-infused and rumen-protected wheat-based pellets containing eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) on n-3 long-chain (≥C₂₀) polyunsaturated fatty acid (LC-PUFA) content, fatty acid recovery, and sensory attributes of ripened cheese from dairy sheep. During a 10-wk supplementary feeding trial, 60 dairy ewes balanced by live weight, milk yield, parity, and sire breed were randomly divided into 6 groups that were (1) supplemented with on-farm existing commercial wheat-based pellets without oil inclusion (control) or supplemented with wheat-based pellets infused with 50 mL/kg dry matter of oils from (2) canola, (3) rice bran, (4) flaxseed, (5) safflower, and (6) rumen-protected EPA + DHA. Milk samples from each treatment were collected separately by sire breed during the experimental period for cheese processing at the end of the experiment. Twelve batches of cheese (2 batches per treatment) were processed and ripened for 120 d. Three cheese samples were collected and analyzed for each cheese making session (total of 36 cheese samples) at d 120 of ripening. Processed cheese of rumen-protected EPA + DHA had the most efficiency at elevating total n-3 LC-PUFA [total EPA + DHA + docosapentaenoic acid (DPA, 22:5n-3] content compared with the control (0.49 vs. 0.28%). Flaxseed elicited the greatest enhancement of α-linolenic acid (ALA, 18:3n-3), whereas safflower was the most effective diet in enhancing the level of linoleic acid (18:2n-6) in cheese (1.29 vs. 0.71% and 4.8 vs. 3.3%, respectively). Parallel recoveries of n-3 and n-6 LC-PUFA were observed across all treatments except for α-linolenic acid and EPA. Cheese eating sensory traits were also highly affected by oil supplementation, with the highest score of 7.5 in cheese from the rice bran and flaxseed treatments. These results provide new insights into the biological mechanisms and processes that determine dairy ewe milk productivity by underpinning the vital biological role of n-3 LC-PUFA in not only enhancing the healthy composition of cheese from ewes but also translating it into consumer acceptability.     

N3系多不饱和脂肪酸膳食参考摄入量的研究进展

近年来n-3系多不饱和脂肪酸（N-3 Polyunsaturated Fatty Acids,n-3 PUFAs）在提高婴幼儿认知及视力发育、增强免疫及预防精神疾病、癌症、炎症、心血管疾病（Cardiovascular Disease,CVD）等方面的作用倍受人们的重视,n-3PUFAs的膳食参考摄入量也成为这些研究领域的重要关注焦点之一。为了更好地发挥n-3 PUFAs在促进人体营养、发育、健康以及疾病预防等方面的重要作用,本文综述了世界上许多知名的研究机构及官方组织关于对不同人群的n-3 PUFAs推荐摄入量（Recommended Nutrient Intake,RNI）及适宜摄入量（Adequate Intake,AI）,以及它们对n-6/n-3脂肪酸的摄入比例以及n-3 PUFAs的可耐受最高摄入量（Tolerable Upper Intake Level,UL）进行的科学研究、探讨,同时还对n-3 PUFAs在使用过程中的不稳定性及注意事项进行了阐述,可以为我国不同人群的n-3 PUFAs膳食参考摄入量、普通食品中应用添加n-3 PUFAs以及n-3 PUFAs摄入补充方面提供更加充分、完整的科学依据和参考。      

Enrichment of eggs in n-3 polyunsaturated fatty acids by feeding hens with different amount of linseed oil in diet

The production of n-3 polyunsaturated fatty acid enriched eggs by addition of linseed oil to the laying hens' diet has been evaluated in terms of production parameters and n-6/n-3 ratio. A total of 150 18weeks old Lohmann Brown laying hens were housed in cages and fed with basal diet and four experimental diets containing 1%, 2%, 3% or 4% of linseed oil added to the basal diet. The effect of the altered level of linseed oil on hens laying performance, fatty acid content and composition and cholesterol content in egg yolk has been evaluated during 13weeks of experiment. Egg weight, yolk fat content, yolk weight, yolk percentage and shape index were not influenced by dietary treatment. The ratio between n-6 and n-3 polyunsaturated fatty acids in eggs decreased in first 5weeks and then remained stable until the end of the experiment for all experimental groups. Different contents of linseed oil in feed highly influenced the n-6/n-3 ratio (P<0.0001). Addition of linseed oil did not influence the cholesterol content in yolks (P=0.5200) while the only factor affecting the cholesterol content was the hens age (P<0.0001).     

Endogenous production of n-3 and n-6 fatty acids in mammalian cells

Polyunsaturated fatty acids (PUFA) are important components of mammalian diets, and the beneficial effects of n-3 PUFA on human development and cardiovascular health have been well documented. Caenorhabditis elegans is one of the few animals known to be able to produce linoleic (LA, 18:2n-6) and alpha-linolenic (ALA, 18:3n-3) essential fatty acids. These essential PUFA are generated by the action of desaturases that successively direct the conversion of monounsaturated fatty acids (MUFA) to PUFA. The cDNA coding sequences of the C. elegans Delta(12) and n-3 fatty acid desaturases were each placed under the control of separate constitutive eukaryotic promoters and simultaneously introduced into HC11 mouse mammary epithelial cells by adenoviral transduction. Phospholipids from transduced cells showed a significant decrease in the ratios of both MUFA:PUFA and n-6:n-3 fatty acids relative to control cultures. The fatty acid profile of transduced cellular phospholipids revealed significant decreases in MUFA and arachidonic acid (20:4n-6), and increases in LA, ALA, and eicosapentaenoic acid (20:5n-3). The fatty acid composition of triacylglycerols derived from transduced cells was similarly, but less dramatically, affected. These results demonstrate the functionality of C. elegans fatty acid desaturase enzymes in mammalian cells. Expression of these desaturases in livestock might act to counterbalance the saturating effect that rumen microbial biohydrogenation has on the fatty acid profile of ruminant products, and allow for the development of novel, land-based dietary sources of n-3 PUFA.