脂肪酶催化棕榈酸和大豆磷脂的酸解反应研究

以大豆磷脂为原料,LipozymeTLIM为酶试剂,正己烷为溶剂,棕榈酸为酰基供体,对大豆磷脂和棕榈酸进行酶法酸解研究。通过实验得出较优的反应条件:液料比(正己烷/磷脂)5∶1,底物摩尔比(棕榈酸/磷脂)5∶1,加酶量(以底物总量为准)20%,反应温度65℃,反应时间60h,水分添加量(以酶量为准)2%,该条件下验证得磷脂中棕榈酸的含量可达到34.45%。     

Interesterification of milkfat with soybean oil catalysed by Rhizopus oryzae lipase immobilised on SiO2‐PVA on packed bed reactor

The potential of the lipase from Rhizopus oryzae immobilised on SiO₂‐PVA to catalyse the interesterification of the milkfat with soybean oil in a packed bed reactor running on continuous mode was evaluated. The reactor operated continuously for 35 days at 45 °C, and during 12 days, no significant decrease in the initial lipase activity was verified. Interesterification yields were in the range from 35 to 38%wt, which gave an interesterified product having 59% lower consistency in relation to non‐interesterified blend. Results showed the potential of the lipase from Rhizopus oryzae to mediate the interesterification of milkfat with soybean oil in packed bed reactor, attaining a more spreadable product under a cool temperature. The biocatalyst operational stability was assessed and an inactivation profile was found to follow the Arrhenius model, revealing values of 34 days and 0.034 day^?1, for half‐life and a deactivation coefficient, respectively.     

无溶剂体系中酶催化合成结构脂质条件初探

以菜籽油和辛酸为原料,在无溶剂体系中用脂肪酶催化酸解合成结构脂质。对6种不同来源的脂肪酶进行筛选,结果表明Lipozyme RMIM催化活性高、Sn-1,3位特异性强。以Lipozyme RMIM为催化用酶,考察了反应时间、反应温度、底物比(菜籽油与辛酸摩尔比)、加酶量、体系水分含量对酸解反应的影响。结果表明,在反应时间15 h,反应温度50℃,底物比1∶4,加酶量10%条件下,辛酸合成率达40%。     

Milk production, conjugated linoleic acid content, and in vitro ruminal fermentation in response to high levels of soybean oil in dairy ewe diet

Feeding vegetable oils rich in linoleic acid has been demonstrated to be an effective strategy to enrich milk with conjugated linoleic acid (CLA). However, high amounts of vegetable oil in the diet in free form could adversely affect animal performance, mainly in sheep. The aim of this work was to improve the ewe milk fatty acid profile by increasing potentially healthy acids such as CLA without any detrimental effects on milk production and ruminal fermentation with soybean oil (SBO) diet supplementation. Twenty-four ewes were assigned to 2 treatments and fed 2 diets (control or supplemented with 6% of SBO; 2 lots of 6 animals per treatment) and fed ad libitum for 4 wk. The forage:concentrate ratio was 20:80. Batch cultures of rumen microorganisms were used to study in vitro rumen fermentation. Changes in fatty acid profile were characterized as a reduction in C6:0 to C16:0 at the expense of an increase in C18:0, C18:1 isomers, and CLA concentrations. Proportions of milk CLA and trans-11 C18:1 (vaccenic acid) went from 1.04 to 3.44 and 2.08 to 6.20 g/100 g of total fatty acids, respectively. However, the SBO diet also increased trans-10 C18:1 and other trans C18:1 content. No significant decreases were found in the treatments for dry matter intake and milk production. The notable increases in trans-10, cis-12 and trans-9, cis-11 were not accompanied by fat level decreases in ewe milk. Concerning in vitro ruminal fermentation, no significant differences were found in the extent and rate of gas production, effective degradability, in vitro true digestibility, and volatile fatty acid production. The results demonstrate that dairy sheep milk CLA content can be substantially increased (more than 3-fold) by adding high levels of SBO in the diet as free oil, without any negative effects on animal performance.     

酶催化桑蚕蛹油酯交换制备中长链脂肪酸甘油酯

以桑蚕蛹油为底物,通过酶法催化其与三辛酸甘油酯进行酯交换反应,制备富含α-亚麻酸的中长链脂肪酸甘油三酯(MLCTs)。通过单因素实验考察了酶种类、底物质量比、反应温度、加酶量以及反应时间对酯交换反应的影响,对酯交换条件进行了优化。结果表明,最佳反应条件为：采用Lipozyme TL IM脂肪酶,三辛酸甘油三酯与桑蚕蛹油质量比1∶ 4,加酶量为底物质量的8%,反应温度45 ℃,反应时间10 h。在最佳反应条件下,酯交换反应的转化率为98.42%,酯交换产物中中长链脂肪酸甘油三酯含量为98.73%,辛酸含量为20.00%,α-亚麻酸含量为30.09%。     

酶催化大豆油酸解制备质构脂质反应条件的优化

探讨了正己烷体系中各因素对酶催化大豆油酸解制备质构脂质反应的影响,在单因素实验基础上,采用响应面设计实验,以辛酸插入率为考察指标,确定了最佳反应条件。结果表明,最优反应条件为:反应温度35.8℃,加酶量11.9%(底物的质量分数),底物摩尔比5.7∶1(辛酸/大豆油),加水量15.4%(酶的质量分数),反应时间20.4 h。在此条件下,实验测得辛酸插入率高达44.903 7%,与模型预测值45.689 8%非常吻合。     

酶法改良大豆油制备质构脂质的研究

研究了固定化脂肪酶催化大豆油与辛酸酸解制备质构脂质的工艺.脂肪酶筛选实验表明,在所选用的三种脂肪酶中,来自Rhizomucor miehei的固定化脂肪酶RM IM催化效果最好.以RM IM为催化剂,进一步考察了酶用量、底物摩尔比、水分添加量、反应温度、反应时间等因素对辛酸插入率的影响.结果表明:当大豆油500mg时,最佳的反应条件为:反应温度40℃,底物摩尔比6∶1(辛酸∶大豆油),固定化酶10%(底物重量百分比),水分添加量10%(酶的重量百分比),反应24h,辛酸的插入率为43%,质构脂质的脂肪酸分布最合理.     

茶油酶促改性一步反应影响因素研究

中国南方几省茶油有一定产量，而国外茶油产量极小，基本上没有市场。利用茶油的特殊结构进行酶促改性制类可可脂很少见有报道。选择１，３位定向脂肪酶，在茶油、硬脂酸、软脂酸混合非水溶剂（己烷）体系中进行一步酯交换反应，最终制出了类似可可脂结构和组成的代用品。本研究主要对茶油酶促改性的主要影响因素进行了探讨。     

利用月桂酸和山茶油的酸解反应分析脂肪酶的位置专一性

脂肪酶的位置专一性在结构酯的合成和油脂改性中具有重要意义。由于水体系和非水体系的差异,传统的水解判定法得出的专一性与该酶在合成反应中的表现可能并不一致。本研究利用月桂酸和山茶油的酸解反应来达到直接评估酶在无溶剂体系中位置专一性的目的。反应产物的脂肪酸组成和位置分布由气相色谱测定。通过此方法,Lipozyme RM IM、L02、L03和L04被鉴定为sn-1(3)位专一性,L01为弱专一性,Novozym 435近似无专一性。通过替换酶的底物,模型反应的可预测性得到验证。根据酸解法和水解法结果的对比分析,2种条件下酶的位置专一性通常是相同的,除了易受到溶剂体系影响的Novozym 435。因此,新方法能够避免水解判定结果在合成反应中应用的局限性。此外,它还降低了对底物纯度的要求,这意味着其能在低成本下用于脂肪酶的大规模筛选。     

Feeding soybean oil to dairy goats increases conjugated linoleic acid in milk

A total of 24 Murciano-Granadina dairy goats milked once daily throughout lactation were used to study the effects of including soybean oil (SBO) in the diet on lactational performance and milk fatty acid (FA) content, particularly conjugated linoleic acid (CLA) and trans-vaccenic acid (trans-11 C18:1, TVA). Three weeks after parturition, goats were allocated to 2 balanced groups according to lactation number, body weight, and daily milk yield, and were kept in separate pens. The experiment consisted of a 2-period (28 d each) crossover with 2 dietary treatments: control and SBO (6% as fed in the concentrate). Goats were fed dehydrated fescue (ad libitum), alfalfa pellets (0.5 kg/d), and concentrate (1 kg/d) to which the SBO was or was not added. Forage was fed in the pens, and concentrate was fed individually in 2 equal portions at milking (0900 h) and in the afternoon (1700 h). Final SBO content in the consumed SBO diet was 2.5% (dry matter basis). Diets were isonitrogenous (17.4% crude protein), but their total FA content varied from 2.2% (control) to 4.6% (SBO). There was no effect of SBO on dry matter intake, milk yield, energy-corrected milk, body weight, or body condition score. Compared with the control diet, feeding SBO increased milk fat content (4.57 vs. 5.24%) and yield as well as total solids content. Soybean oil had no effect on milk crude and true protein contents, but it reduced milk casein content (2.48 vs. 2.34%). Short- and medium-chain FA decreased by feeding SBO, whereas long-chain FA increased. Feeding preformed linoleic acid through SBO increased milk concentrations of linoleic, oleic, and stearic FA but reduced levels of linolenic and palmitic FA. As a consequence, feeding SBO decreased the saturated-to-unsaturated FA ratio and the atherogenicity index. Compared with the control treatment, milk contents of cis-9, trans-11 CLA (0.68 vs. 2.03%) and TVA (2.04 vs. 6.41%) in the SBO treatment increased by approximately 200%. In conclusion, feeding a moderate dose of SBO to dairy goats was a useful way to increase milk fat, CLA, and TVA contents in milk and to reduce the atherogenicity index without negative effects on intake, milk yield, and protein content.     

酶法合成高含量共轭亚油酸甘油酯

选用AB-8大孔弱极性树脂对脂肪酶进行固定化,固定化脂肪酶的活力为210U／g。用此固定化酶催化共轭亚油酸乙酯和大豆油合成富含共轭亚油酸（CLA）的改性大豆油,最佳反应条件为：底物摩尔比（n（CLA乙酯）：n（大豆油））1：0．33,酶加量21U／g,反应温度60℃。放大反应体系,对反应产物中的CLA含量进行检测分析表明,其含量可达37％。     

Production tactic and physiochemical properties of low ω6/ω3 ratio structured lipid synthesised from perilla and soybean oil

Structured lipid (SL) was synthesised by enzymatic interesterification of soybean oil and perilla oil. Response surface methodology (RSM) was used to determine the effects of three variables on the lipase‐catalysed interesterification. Based on ridge analysis, combination of reaction time (X₁; 18 h), reaction temperature (X₂; 60 °C), and substrate mole ratio (X₃; 1:1) were optimised for higher incorporation of ω3 (alpha (α) linolenic acid (ALA)) (Y). The predictive model was found to be adequate due to no significant lack of fit and satisfactory level of coefficient of determination (R² = 0.97). Experiments were conducted under optimised conditions which were predicted by the model equations, obtained from RSM yielded SL with linoleic (44.01%) and ALA (35.82%) were detected at sn‐2 position. The effects of antioxidants such as catechin, butylated hydroxytoluene (BHT) BHT and rosemary extracted on the oxidative stability in SL were investigated. Among all antioxidants, the highest stability was obtained from catechin.     

无溶剂体系酶法催化合成共轭亚油酸薄荷酯

在无溶剂体系中采用脂肪酶AY 30催化共轭亚油酸(CLA)和L-薄荷醇反应,合成共轭亚油酸薄荷酯。研究了酶用量、水用量、反应温度和反应时间对酯化率和产物共轭亚油酸薄荷酯组成的影响。结果表明:最佳酯化率的反应条件为酶用量2%,水用量8%,反应温度45℃,反应时间48h;产物中c9t,11-CLA含量最高时的反应条件为酶用量2%,水用量4%,反应温度40℃,反应时间12 h,在此条件下,产物中c9,t11-CLA含量可达86.32%。     

棕榈油硬脂和大豆油酶法酯交换的研究

进行了固定化脂肪酶Lipozyme TL IM催化棕榈油硬脂和大豆油酶法酯交换的研究,考察了酶法酯交换分批操作的反应动力学曲线,发现5%的加酶量、70℃下反应,反应3 h内达到平衡,反应温度在70~90℃对反应速度无明显影响。采用填充床式反应器进行连续反应,结果表明通过装有10 g脂肪酶柱子的最佳流速为30 g/h。对化学酯交换和酶法酯交换产品性质的比较发现,二者SFC无明显差异。采用填充床式反应器对40∶60和30∶70的棕榈油硬脂和大豆油的混合物进行酶法酯交换反应,测定了反应产物的SFC曲线,为将来的应用开发提供参考数据。     

响应面优化脂肪酶催化酸解合成共轭亚油酸磷脂

采用商业化脂肪酶Lipozyme RM IM作为生物催化剂,催化共轭亚油酸(CLA,80％)和大豆磷脂( PC90)酸解反应合成富含CLA的结构磷脂.利用响应面分析方法研究了在正己烷溶剂体系中,底物摩尔比、酶用量、反应温度和反应时间对产物中CLA含量的影响.通过分析验证得到最佳反应条件为∶CLA与大豆磷脂的摩尔比6∶1,酶用量30％(以底物总质量计),反应温度48℃,反应时间64 h.在最佳反应条件下,产物中CLA的含量为24.18％.     

超声强化镍催化油脂共轭反应的研究

研究了超声强化镍催化剂催化油脂共轭反应的效果.在超声作用下,以大豆油为原料,镍为固体催化剂,氮气为保护气,通过单因素和正交实验,分别考察了超声功率、超声作用方式、反应温度、反应时间、催化剂用量等因素对油脂共轭反应的影响.结果表明最佳工艺条件为:超声功率25W,超声作用方式5 s/7 s,催化剂用量10％,反应时间50 min,反应温度140℃.在最佳工艺条件下得到油脂共轭亚油酸含量为7.498 mg/mL.     

酶法制取低热量大豆功能性油脂的研究

以大豆油和丙酸为实验原料,采用无溶剂体系,酶法制备低热量功能性油脂。结果表明,固定化1-3定向脂肪酶LipozymeRM IM比Li-pozyme TL IM具有较高的催化活性。根据Box-Behnken中心组合实验设计原理,确定了制备低热量功能性油脂的最佳反应条件为:温度64.4℃,时间8.8h,酶添加量8.4%,底物摩尔比3.3∶1。在此最佳反应条件下,丙酸插入率可达14.85%。      

大豆油脱臭馏出物酶法甲酯化工艺研究

Eversa Transform 2. 0脂肪酶价格低、催化活性好,具有微量高效的优点。以Eversa Transform2. 0脂肪酶为催化剂,对大豆油脱臭馏出物进行酶法甲酯化工艺研究。通过单因素试验研究了醇酸摩尔比、酶用量、反应温度、反应时间对酯化率的影响,并进行正交试验,对酶法甲酯化工艺进行优化。在最佳工艺条件下,通过放大试验对试验结果进行验证,并采用薄层色谱法对甘油三酯的转化进行分析。结果表明:Eversa Transform 2. 0脂肪酶催化大豆油脱臭馏出物甲酯化反应的最佳工艺条件为醇酸摩尔比(甲醇与游离脂肪酸摩尔比) 2∶1、酶用量0. 3%(相对于大豆油脱臭馏出物的质量)、反应温度35℃、反应时间6 h,在此条件下酯化率达到95%以上,且甘油三酯基本反应完全。