发酵法生产多价不饱和脂肪酸的研究

γ-亚麻酸在医药、食品、化妆品和饲料领域拥有广阔的应用前景,研究发现采用小于16％糖浓度的流加工艺进行发酵时,γ-亚麻酸产量为2.4g/L,生产周期短,生产消耗低,适合实际生产应用,适宜的产物提取方法是乙醇-正己烷两步法;DHA、EPA等不饱和脂肪酸主要结合在甘油的2-位,利用脂肪酶对油脂选择性水解的特点对鱼油进行水解反应可以达到富集多价不饱和脂肪酸的目的,筛选了两种目标酶并研究了其固定化方法;用被孢霉属菌株在100L发酵罐中生产花生四烯酸,菌干重可达36g/L,花生四烯酸达4.9g/L。     

卤虾油的醇解工艺

利用正交试验研究了酯交换法制备卤虾油脂肪酸乙酯的工艺,分析了催化剂用量、反应温度、反应时间、无水乙醇用量等对醇解率的影响.结果表明：当乙醇钠催化剂为卤虾油质量的0.5%,反应温度60℃,反应时间3 h,乙醇和卤虾油质量比为0.40的条件下,卤虾油的醇解率可达98.04%.从而为使卤虾油替代鱼油成为保健品行业的原材料提供依据.     

尿素包合法富集鱼油中的EPA和DHA的工艺研究

运用尿素包合法富集鱼油中的EPA和DHA。考察了尿脂比、反应温度、结晶温度和结晶时间对EPA和DHA总纯度的影响。随尿脂比的增大,总纯度先增大后趋于稳定,适宜的尿脂比为2．5：1。随反应温度的升高,总纯度逐渐增大,适宜的反应温度为55℃。随结晶温度的降低,总纯度逐渐增大,适宜的结晶温度为0℃。随结晶时间的增大,总纯度先增大后趋于稳定,适宜的结晶时间为1h。     

多烯脂肪酸乙酯的制备与提纯

本文在传统工艺的基础上,对鱼油中的多烯脂肪酸进行酯化,并对其酯化产物的提纯提出新的工艺路线即采用尿素包合法与高真空精馏法相结合的方法,以克服原有工艺中的有不足,提高产品质量。     

毛细管气相色谱法测定鱼油脂肪乳注射液中DHA和EPA的含量

采用直接进样毛细管气相色谱法测定鱼油脂肪乳注射液中DHA和EPA的含量,色谱柱为PEG-20M毛细管柱,柱温采用程序升温,载气为氦气,流速0.5 mL/min,采用FID检测器,温度为280℃,进样口温度300℃,分流比5∶1,进样量1μL。结果表明,DHA甲酯及EPA甲酯浓度分别为0.197～0.690 g/L 0,.208～0.743 g/L时线性关系良好r,分别为0.999 50,.998 4;平均回收率分别为99.99%9,9.41%,RSD分别为0.52%,0.47%。该法简便、准确,重现性好,可用于鱼油脂肪乳注射液中DHA和EPA的含量测定。     

分子筛吸附鱼油中DHA与EPA特性的研究

实验改进了硝酸银修饰分子筛的方法.研究了修饰分子筛对EPA乙酯和DHA乙酯的吸附特性.结果表明,分子筛对EPA乙酯和DHA乙酯的吸附属于单分子层吸附.修饰分子筛在40℃时对EPA乙酯和DHA乙酯的最大吸附量分别为37.4 mg/g和24.2 mg/g.     

DHA合成条件探究

以乙酰乙酸乙酯为原料合成脱氢醋酸,并通过熔点测定、IR、MS、1HNMR对产物进行简单的表征。探讨了催化剂的种类和用量、反应时间以及反应温度等对产物产率的影响。结果表明：当选用碳酸氢钠做催化剂合成DHA时,催化剂用量为0.11%,反应釜温度控制在180℃～190℃,时间为120 min,产物产率最高为51.3%,所得产物为浅黄色晶体;通过薄层色谱分析,所含杂质少。     

Enzymatic and chemical synthesis of phosphatidylcholine regioisomers containing eicosapentaenoic acid or docosahexaenoic acid

Regioselective incorporation of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) into phosphatidylcholine (PC) was carried out using enzymatic and chemical synthesis. Incorporation at the sn‐1 position was successfully achieved by lipase‐catalysed esterification of 2‐palmitoyl‐lysophosphatidylcholine (LPC), although in most cases, the enzymes incorporated EPA and DHA at lower rates than other fatty acids. For the incorporation of DHA, Candida antarctica lipase B was the only useful enzyme, while incorporation of EPA was efficiently carried out using either this enzyme or Rhizopus arrhizus lipase. The highest yields in the lipase‐catalysed reactions were obtained at the lowest water activity (close to 0). However, by carrying out the reactions at a higher water activity of 0.22, more EPA and DHA were incorporated. Esterification of 2‐palmitoyl‐LPC with pure EPA at this water activity converted 66 mol‐% of LPC to PC using Rhizopus arrhizus lipase as catalyst. When the fatty acid was DHA and the catalyst Candida antarctica lipase B, 45 mol‐% of PC was obtained. For incorporation of EPA and DHA at the sn‐2 position, phospholipase A₂ was used, but the reaction was very slow. Chemical coupling of 1‐palmitoyl‐LPC and EPA or DHA was more efficient, resulting in complete conversion of LPC.     

Oxidative Stability of Liposomes Composed of Docosahexaenoic Acid-Containing Phospholipids

Oxidative stability of liposomes made of (Docosahexaenoic acid) DHA-containing phosphatidylcholine (PC) was examined during preparation and storage. After preparation of the liposomes, the concentration of primary (conjugated dienes) and secondary oxidation products (Thiobarbituric acid-reactive substances, TBARS) were significantly higher compared to the initial value. During cold storage, formation of conjugated dienes and TBARS remained more or less constant in large unilamellar vesicles (LUV), whereas in mulitilamellar vesicles (MLV) they were seen to increase over a period of 21 days. Evaporation of solvent traces from a lipid film should preferably be done under nitrogen as vacuum evaporation was found to increase oxidation of the phospholipid.     

超临界CO2流体从紫菜中萃取EPA和DHA的研究

采用超临界CO2 流体萃取技术从紫菜中萃取EPA和DHA ,研究了紫菜粉碎粒度、萃取温度、萃取压力和萃取时间对萃取率的影响。结果表明 :超临界CO2 流体从紫菜中萃取EPA和DHA的工艺可行 ,最佳的萃取条件为 :紫菜粉碎粒度为 2 0～ 80目 ,萃取温度 3 5℃ ,萃取压力 2 5MPa ,萃取时间 1 .5h。     

高纯度EPA/DHA甘油三酯的酶法合成

利用脂肪酶分别催化游离型EPA和DHA与甘油发生酯化反应生成甘油三酯,考察了合成的影响因素. 结果表明,正己烷6 mL,甘油/EPA(DHA)摩尔比为1:3(以0.4 mmol甘油为基准),Novozym 435添加量为100 mg,反应温度40℃,振荡频率150 r/min,反应24 h后添加1 g分子筛,反应时间48 h,EPA甘油三酯与DHA甘油三酯的得率分别可达88.64%和88.07%,EPA与DHA酯化度分别可达95.0%和94.5%. 分析结果表明,所得产物为EPA甘油三酯和DHA甘油三酯.     

DHA and Its Metabolites Have a Protective Role against Methylmercury-Induced Neurotoxicity in Mouse Primary Neuron and SH-SY5Y Cells

The consumption of fish now involves a risk of methylmercury (MeHg) exposure but also provides the benefit of ω-3 polyunsaturated fatty acids (ω-3 PUFAs) such as docosahexaenoic acid (DHA). Some epidemiological studies have suggested that the intake of DHA can alleviate the neurotoxicity of MeHg, but the underlying mechanism is not known. Herein, we observed that pretreatment with 0.1–1 µM DHA suppressed MeHg-induced cytotoxicity in human neuroblastoma (SH-SY5Y) cells and mouse primary neuronal cells. These effects of DHA were canceled in the presence of the retinoid X receptor (RXR) antagonist UVI3003. An RXR agonist, bexarotene, suppressed the cytotoxicity of MeHg. DHA also suppressed the MeHg-induced production of reactive oxygen species (ROS) via an induction of antioxidant genes (catalase and SOD1). Pretreatment with DHA did not change the incorporation of MeHg. We showed previously that in the brain, the intake of DHA increased the level of 19,20-DHDP, which is the metabolite produced by cytochrome P450 and soluble epoxide hydrolase from DHA. In the present study, we observed that 19,20-DHDP also suppressed neurotoxicity from MeHg. These results indicate that DHA and its metabolites have a protective role in MeHg-induced neurotoxicity.     

超临界CO2从螺旋藻中萃取EPA和DHA工艺研究

研究采用超临界ＣＯ２从螺旋藻中萃取ＥＰＡ和ＤＨＡ的最优化工艺条件，得最高萃取率ＥＰＡ为９３％，ＤＨＡ为８９．３％，该法比溶剂法优越，为开发和综合利用海藻资源开辟了新的应用途径。     

国内EPA及DHA研究现状和发展趋势

本文对国内有关EPA（Eicosapentaenoicacid）及DHA（Docosahexaenoicacid）的特性、分离纯化方法及应用等研究现状进行了评述，并指出了与国际发达国家的差距，提出了今后工作的建议．     

Artesunate and dihydroartemisinin (DHA): unusual decomposition products formed under mild conditions and comments on the fitness of DHA as an antimalarial drug

Artesunate drug substance, for which a rectal capsule formulation is under development for the treatment of severe malaria, when heated at 100 degrees C for 39 h gives beta-artesunate, artesunate dimers, 9,10-anhydrodihydroartemisinin (glycal), a DHA beta-formate ester, and smaller amounts of other products that arise via intermediate formation of dihydroartemisinin (DHA) and subsequent thermal degradation. Solid DHA at 100 degrees C provides an epimeric mixture of a known peroxyhemiacetal, arising via ring opening to a hydroperoxide and re-closure, smaller amounts of a 3:1 mixture of epimers of a known tricarbonyl compound, and a single epimer of a new dicarbonyl compound. The latter arises via homolysis of the peroxide and an ensuing cascade of alpha-cleavage reactions which leads to loss of formic acid incorporating the C10 carbonyl group of DHA exposed by this 'unzipping' cascade. The tricarbonyl compound that arises via peroxide homolysis and extrusion of formic acid from a penultimate hydroxyformate ester incorporating C12 of the original DHA, is epimeric at the exocyclic 1'-aldehyde, and not in the cyclohexanone moiety. It is converted into the dicarbonyl compound by peroxide-induced deformylation. The dicarbonyl compound is not formed during anhydrous ferrous bromide mediated decomposition of DHA at room temperature, which provides the 1'-R epimer of the tricarbonyl compound as the dominant product; this equilibrates at room temperature to the 3:1 mixture of epimers of the tricarbonyl compound obtained from thermolysis. Each of artesunate and DHA decomposes readily under aqueous acidic conditions to provide significant amounts of the peroxyhemiacetal, which, like DHA, decomposes to the inert end product 2-deoxyartemisinin under acidic or basic conditions. DHA and the peroxyhemiacetal are the principal degradants in aged rectal capsule formulations of artesunate. TGA analysis and thermal degradation of DHA reveals a thermal lability which would pose a problem not only in relation to ICH stability testing guidelines, but in the use of DHA in fixed formulations currently under development. This thermolability coupled with the poor physicochemical properties and relative oral bioavailability of DHA suggests that it is inferior to artesunate in application as an antimalarial drug.     

Genomic Analysis of Genes Involved in the Biosynthesis of Very Long Chain Polyunsaturated Fatty Acids in Thraustochytrium sp. 26185

Thraustochytrium sp. 26185 is a marine protist that can produce a large amount of docosahexaenoic acid (DHA, 22:6n‐3), an ω3 very long chain polyunsaturated fatty acid (VLCPUFA) of nutritional importance. However, the mechanism of how this fatty acid is synthesized and assembled into the storage lipid triacylglycerol is unclear. Here we report sequencing of the whole genome and genomic analysis of genes involved in the biosynthesis and assembly of the fatty acids in this species. Genome sequencing produced a total of 2,418,734,139 bp clean sequences with about 62 fold genome coverage. Annotation of the genome sequences revealed 10,797 coding genes. Among them, 10,216 genes could be assigned into 25 KOG classes where 451 genes were specifically assigned to the group of lipid transport and metabolism. Detailed analysis of these genes revealed co‐existence of both aerobic pathway and anaerobic pathways for the biosynthesis of DHA in this species. However, in the aerobic pathway, a key gene encoding stearate Δ9 desaturase introducing the first double bond to long chain saturated fatty acid 18:0 was missing from the genome. Genomic survey of genes involved in the acyl trafficking among glycerolipids showed that, unlike plants, this protist did not possess phosphatidylcholine:diacylglycerol cholinephosphotransferase, an important enzyme in bridging two types of glycerolipids, diacylglycerols (DAG) and phosphatidylcholines (PtdCho). These results shed new insight on the biosynthesis and assembly of VLCPUFA in the Thraustochytrium.     

Strategies for the enzymatic enrichment of PUFA from fish oil

PUFA from oil extracted from Nile perch viscera were enriched by selective enzymatic esterification of the free fatty acids (FFA) or by hydrolysis of ethyl esters of the fatty acids from the oil (FA‐EE). Quantitative analysis was performed using RP‐HPLC coupled to an evaporative light scattering detector (RP‐HPLC‐ELSD). The lipase from Thermomyces lanuginosus discriminated against docosahexaenoic acid (DHA) most, resulting in the highest DHA/DHA‐EE enrichment while lipase from Pseudomonas cepacia discriminated against eicosapentaenoic acid (EPA) most, resulting in the highest EPA/EPA‐EE enrichment. The lipases discriminated between DHA and EPA with a higher selectivity when present as ethyl esters (EE) than when in FFA form. Thus when DHA/EPA were enriched to the same level during esterification and hydrolysis reactions, the DHA‐EE/EPA‐EE recoveries were higher than those of DHA/EPA‐FFA. In reactions catalysed by lipase from T. lanuginosus, at 26 mol% DHA/DHA‐EE, DHA recovery was 76% while that of DHA‐EE was 84%. In reactions catalysed by lipase from P. cepacia, at 11 mol% EPA/EPA‐EE, EPA recovery was 79% while that of EPA‐EE was 92%. Both esterification of FFA and hydrolysis of FA‐EE were more effective for enriching PUFA compared to hydrolysis of the natural oil and are thus attractive process alternatives for the production of products highly enriched in DHA and/or EPA. When there is only one fatty acid residue in each substrate molecule, the full fatty acid selectivity of the lipase can be expressed, which is not the case with triglycerides as substrates.     

工业DHA微胶囊物理性质的表征

二十二碳六烯酸（DHA）是一种对人体非常重要的多元不饱和脂肪酸。目前,有关DHA微胶囊物理性质的表征有限。本文从DHA微胶囊的表观形态、粒径分布、堆积密度、孔径分布、比表面积、流动性、喷流性、复水性和热力学性质等几个方面对所选定的典型工业DHA微胶囊进行表征。实验结果表明,DHA微胶囊的震实密度会随震动次数、震动频率、震动幅度的增加而增加;在震动次数、频率和震幅分别达到1000次、112min。和9mm之后趋于稳定。采用carr流动指数法,通过加权计算,结果表明该DHA微胶囊流动性质一般,喷流性很强。通过显微镜观察DHA微胶囊复水过程中的形态变化,表明此DHA微胶囊复水性质良好。     

Bioavailability of EPA and DHA delivered by gelled emulsions and soft gel capsules

This study presents data comparing the bioavailability of DHA and EPA delivered by two different formulations: One group received TAG fish oil in traditional soft gel capsules, whereas the other group received the TAG oil as droplets trapped inside a gelatin matrix (gelled emulsions). The incremental area under the curve (AUC_{0–26 h}) of EPA and EPA + DHA in blood plasma from the gelled emulsions was significantly increased by 44.9 and 43.3%, respectively, compared to soft gel capsules. The maximum incremental concentration of EPA and EPA + DHA was significantly increased by 100.4 and 105.6%, respectively, compared to soft gel capsules. These results suggest that improved bioavailability of EPA and DHA may be achieved by incorporating emulsified TAG fish oil in a gel matrix prior to oral ingestion. Practical applications: This study presents a new type of vehicle for the delivery of PUFAs. The vehicles are soft and chewable with the possibility of adding flavours, sweeteners and colour, and this makes the vehicles ideal for delivery of PUFAs to consumers having problems swallowing large capsules or cod liver oil. The vehicles are already applied in products in several countries, including Norway.