酶催化合成蔗糖酯研究进展

本文对蔗糖酯的酶促合成研究进行了总结,其中主要涉及到酶、反应媒介和酰基供体等影响因素。研究发现,蛋白酶、脂肪酶和抗体酶均能区域选择地酰化蔗糖。酶催化合成蔗糖酯通常在N,N-二甲基甲酰胺(DMF),二甲基亚砜(DMSO)和吡啶等有毒溶剂中进行。近几年来,使用混合溶剂或叔丁醇等毒性较小的溶剂代替毒性较大的溶剂,以及无溶剂条件下酶催化合成蔗糖酯的研究,也越来越受重视。活化酯作为酰基供体得到广泛的应用。     

锦葵啶葡萄糖苷酶促酰化反应体系的研究及效果评价

试验旨在建立锦葵啶葡萄糖苷酶促酰化反应的反应体系,探究酶促酰化反应的影响因素;探讨花色苷酶促酰化反应产物的亲脂性和稳定性.以酶活为指标筛选出最佳反应介质,以花色苷的残留率和色差确定最适合花色苷酶促酰化反应的脂肪酶、最佳酶添加量、最佳分子筛添加量和酰基供体.结果表明:二甲基四氢呋喃(2-MeTHF)为最佳反应介质;Candila antarctica脂肪酶B(CAL-B,Novozym 435)为最佳添加酶试剂;肉桂酸乙烯酯为最佳酰基供体;当反应体系加入2 mL脱水2-MeTHF、0.01 mmol锦葵啶葡萄糖苷、0.15 mmol肉桂酸乙烯酯时,酶和分子筛的最佳添加量分别为50 U和0.15 g.按照试验所建立的酶促酰化反应体系完成锦葵啶葡萄糖苷的酰化反应,对比酰化前后试样的紫外-可见光谱图,结果显示生成了酰化锦葵啶葡萄糖苷,证明了酶促酰化处理具有酰化效果.酰化反应后花色苷的亲脂性和稳定性显著提高,可以推断酰化是使花色苷稳定性提高的根本原因.研究结果可为花色苷结构稳定化提供借鉴.     

一种深海微生物脂肪酶在非水介质中催化合成乙酸肉桂酯的应用研究

从一株深海来源微生物Streptomyces sp.SCSIO 13580的基因组中克隆了1个序列新颖的脂肪酶编码基因,含有1005个碱基,编码334个氨基酸,与最相近的蛋白有51%相似。在大肠杆菌中异源表达了这个脂肪酶,发现其具有在非水介质中催化合成乙酸肉桂酯的活性。在单因素实验的基础上,考察了酰基供体、溶剂、肉桂醇/乙酸乙烯酯摩尔浓度之比、温度等因素对该酶合成乙酸肉桂酯的活性的影响,得到的最佳反应条件是:以乙酸乙烯酯为酰基供体,以正己烷为溶剂,肉桂醇/乙酸乙烯酯的摩尔浓度之比为1∶6,反应温度为40℃,优化后的转化率达到48.3%,酶粉的酶活力为5.59 U/g。     

重组脂肪酶全细胞催化单糖酯的合成

利用毕赤酵母表面展示南极假丝酵母脂肪酶B(Candida antarctica lipaseB,CALB)为全细胞催化剂,以不同单糖(葡萄糖、果糖和甘露糖)为酰基受体,不同脂肪酸(月桂酸、油酸和棕榈酸)为酰基供体,在以不同有机溶剂为介质的非水相体系中催化合成糖酯.探讨全细胞脂肪酶CALB合成糖酯的反应条件,结果表明:毕赤酵母表面展示CALB全细胞催化剂在叔戊醇/二甲基亚砜(4:1,v/v)介质中可以有效的合成糖酯,研究中各种酰基受体和酰基供体在全细胞脂肪酶催化下均能合成糖酯,其中果糖作为酰基受体产物浓度最高.     

酶促酸解合成结构脂反应中酰基迁移的研究进展

酶促酸解合成结构脂具有显著的优越性,其中酰基迁移副反应是结构脂合成中亟需研究和控制的关键。对酶催化酸解反应中酰基迁移的理论和实践研究进行综述,归纳总结温度、时间、酶、载体、水分活度、溶剂、底物比、甘油酯结构等因素对酰基迁移的影响。目前,对酰基迁移的研究主要在酶促反应水平上,机理尚不深入,且优化酶促酸解条件提高接入率、产率与减少酰基迁移是不可兼得的矛盾关系。今后需要进行酰基迁移在复杂体系下的影响机制和评测方法的深入研究,以期更高效地开发特殊用途的结构脂产品。     

花色苷酶促酰化反应研究进展

花色苷的功能、稳定性与其结构存在密切关联,特别是其酰基结构。酰化反应是提高花色苷结构稳定性的重要方法。传统化学酰化方法存在许多不足和缺陷,而酶促酰化受到广泛关注,因其反应条件温和,选择性强,产品质量高,可运用在食品,药品,化工合成等领域。总结了酰化反应对提高花色苷稳定性的效用,酶促糖苷酰化反应中催化酶、酰基供体以及溶剂体系方面的研究进展,以期对花色苷结构稳定化技术提供借鉴。     

表没食子儿茶素没食子酸酯的化学修饰及生物活性研究进展

表没食子儿茶素没食子酸酯(epigallocatechin gallate, EGCG)是茶叶中的一种重要儿茶素,具有良好的生物活性,但口服生物利用度低、脂溶性及稳定性差等缺点使其在功能性健康产品开发中的应用受到限制。研究发现,通过醚化、酯化和糖苷化等化学修饰方法,在EGCG结构中引入烷基、酰基、糖基等基团可得到不同EGCG衍生物,显著改善EGCG的理化性质,并特异性增强EGCG的抗氧化、抗炎、抗病毒、抗衰老、抗癌等生物活性。醚化修饰的操作简便,但产物复杂、不利于后续的分离纯化;酯化修饰的研究多集中在小分子试剂上;糖苷化修饰的合成技术难度较高,但是该修饰方法可定向制备目标产物。本文整理近几年的国内外相关文献,对EGCG化学修饰的常见方法及EGCG衍生物的活性研究进行了系统的总结,为EGCG的构效关系研究及开发EGCG类功能性健康产品提供参考。     

脂肪酶催化结构酯的合成及其应用概况

综述通过脂肪酶催化特异性作用合成结构酯产物(specific structured triacylglycerols，SSTs)，同时对合成SSTs的反应底物、脂肪酶和酰基供体，反应类型及其影响因素以及应用概况进行了较为详尽的介绍。     

高油酸花生油酶促酯交换合成类可可脂工艺优化

为促进高油酸花生油的高值化利用,在无溶剂体系中以高油酸花生油为原料,棕榈酸乙酯、硬脂酸乙酯为酰基供体,酶促酯交换合成类可可脂。以目标甘三酯1,3-二棕榈酸-2-油酸(POP)、1-棕榈酸-2-油酸-3-硬脂酸(POS)、1,3-二硬脂酸-2-油酸(SOS)含量,硬脂酸指数和酰基位移率为评价指标,在单因素实验的基础上采用响应面法对酶促酯交换合成类可可脂的工艺条件进行优化。结果表明：酶促酯交换合成类可可脂的最佳工艺条件为硬脂酸乙酯与棕榈酸乙酯物质的量比1.3∶1、酰基供体与高油酸花生油物质的量比12∶1、加酶量2.7%(以底物质量计)、反应温度60℃、反应时间7 h,在该条件下产物甘三酯中POP、POS、SOS的含量分别为14.55%、48.87%、25.17%,硬脂酸指数为0.56,酰基位移率为7.35%。产物的目标甘三酯组成和脂肪酸组成与可可脂相近,可作为可可脂替代品应用。     

无溶剂体系中毕赤酵母表面展示CALB全细胞催化合成肉桂醇酯

利用表面展示南极假丝酵母脂肪酶B(Candida antarctica lipase B,CALB)的毕赤酵母细胞(Pp-CALB)为全细胞催化剂,以不同脂肪酸作为酰基供体,在无溶剂体系中和肉桂醇发生酯化反应合成肉桂醇酯。探究了Pp-CALB对于不同酰基供体的选择性,并且对反应温度、摇床转速、酶加量、底物摩尔比、酶的水活度和反应体系等一系列影响酶催化反应的因素进行了优化,建立了无溶剂体系中肉桂醇酯的酶法合成工艺。结果表明,在底物摩尔比为2:1,摇床转速为120 r/min,反应温度为50℃的条件下,添加初始水活度为0.53的Pp-CALB 0.05 g,反应3h,肉桂醇的酯化率可达81.34%,在最适反应条件下,使用Pp-CALB连续反应8次后,肉桂醇的酯化率仍然能达到70.00%以上,具有较好的操作稳定性。     

Antioxidant properties of modified rutin esters by DPPH,reducing power,iron chelation and human low density lipoprotein assays

Practical limitations exist regarding the effectiveness of flavonoids as antioxidants in many food systems, possibly due to their poor solubility and miscibility in lipidic environments. Current strategies to improve these properties include enzymatically acylating flavonoids with lipophilic moieties. Herein, two derivatives of rutin (possessing C12:0 or C16:0 acyl groups) were assessed for their antioxidant properties, and compared with their parent compound, rutin and with butylated hydroxytoluene (BHT). While all compounds exhibited relatively strong radical scavenging abilities, modified rutin compounds exhibited decreased reducing power and metal chelating abilities as compared to rutin. Conversely, investigations on the oxidation of human low density lipoprotein (LDL) revealed that rutin laurate was most effective in inhibiting oxidation by prolonging LDL lag time for an in vitro system. With regards to in vivo considerations, a pre-treatment step confirmed that the ester bond linking rutin and acyl moieties was most susceptible to hydrolysis by digestive enzymes, while rutin itself was not degraded. Thus, acylation of rutin with medium or long chain fatty acids may result in improved antioxidant abilities in more complex systems, including LDL-oxidation assays. Likely reasons may include improved lipophilic solubility and partitioning properties allowing for better accessibility to the actual site of oxidation.     

Loss of rutin and antioxidant activity of asparagus juice caused by a pectolytic enzyme preparation from Aspergillus niger

We found that a commercial pectolytic enzyme preparation from Aspergillus niger (pectinase AN) contained laccase activity that decreased rutin content and antioxidant activity of asparagus juice. This research investigated the effects of pH, temperature, and concentration of pectinase AN on pectinase AN’s laccase activity to decrease rutin content and antioxidant activity of asparagus juice. Asparagus juice was incubated with pectinase AN at different pHs (3.2, 4.5 and 5.8), temperatures (25, 37, and 50 °C) and enzyme concentrations (0.1%, 0.5% and 1%). Rutin content and antioxidant activity of samples was determined by HPLC and 2,2′-diphenyl-1-picrylhydrazyl (DPPH) free radical method, respectively. The rate of loss of rutin and antioxidant activity of asparagus juice was smaller at pH 3.2 than at pH 4.5 and pH 5.8, smaller for 0.1% pectinase AN than 0.5% and 1% pectinase AN. The rate of loss of rutin of asparagus juice was greater at 25 °C than at the other two temperatures. Pectinase AN can decrease rutin content and antioxidant activity of asparagus juice at the selected conditions. But rutin content and antioxidant activity of asparagus juice produced using pectinase AN could be less decreased at pH 3.2 and 0.1% of enzyme with less than 2 h of incubation time. This information was helpful for juice industry to produce juices with high antioxidant activity using pectinase AN.     

Subcritical Water Induced Complexation of Soy Protein and Rutin: Improved Interfacial Properties and Emulsion Stability

Rutin is a common dietary flavonoid with important antioxidant and pharmacological activities. However, its application in the food industry is limited mainly because of its poor water solubility. The subcritical water (SW) treatment provides an efficient technique to solubilize and achieve the enrichment of rutin in soy protein isolate (SPI) by inducing their complexation. The physicochemical, interfacial, and emulsifying properties of the complex were investigated and compared to the mixtures. SW treatment had much enhanced rutin‐combined capacity of SPI than that of conventional method, ascribing to the well‐contacted for higher water solubility of rutin with stronger collision‐induced hydrophobic interactions. Compared to the mixtures of rutin with proteins, the complex exhibited an excellent surface activity and improved the physical and oxidative stability of its stabilized emulsions. This improving effect could be attributed to the targeted accumulation of rutin at the oil–water interface accompanied by the adsorption of SPI resulting in the thicker interfacial layer, as evidenced by higher interfacial protein and rutin concentrations. This study provides a novel strategy for the design and enrichment of nanovehicle providing water‐insoluble hydrophobic polyphenols for interfacial delivery in food emulsified systems.     

Computational and experimental validation of antioxidant properties of synthesized bioactive ferulic acid derivatives

Ferulic acid, a well-known natural phenolic compound, is considerably reported for its hydroxyl and peroxyl radical scavenging activities. However, the antioxidant capacity of ferulic acid is limited by its relatively low solubility in hydrophobic media thereby preventing its application for autoxidation of fats and oils. To this end, several research efforts are being made to maximize the therapeutic benefits of ferulic acid and these efforts included but not limited to structural modification of ferulic acid to produce its derivatives. In this study, we synthesized ester and amide derivatives of ferulic acid and evaluated them for in vitro antioxidant potential as well as molecular docking properties. Data revealed that ferulic ester and amide derivatives had excellent antioxidant capacity and demonstrated strong inhibitory potential. Analysis of molecular docking indicated ferulic ester as potent inhibitor of target proteins in breast cancer as well as in oxidative stress. Taken together, the findings support potent antioxidant properties by these ferulic derivatives. Findings may become relevant where structural modification to enhance physico-chemical properties without compromising the antioxidant and/or medicinal potential are desirable.     

酶法改性影响膳食纤维的构成及生物作用效果的研究进展

膳食纤维根据溶解性分为可溶性膳食纤维与不溶性膳食纤维。一些食品中可溶性膳食纤维的含量低限制了其在食品工业中的应用,因此将不溶性膳食纤维转变成可溶性膳食纤维的改性方法成为人们研究的目标。众多方法之中,在添加外源酶的作用下对不溶性膳食纤维进行酶法改性可以有效地增加膳食纤维溶解性。酶法改性后,膳食纤维的单糖组成、官能团、结构与结晶度等构成和降胆固醇、降血糖、抗氧化、离子交换能力等生物作用效果均发生变化,可以提高其应用价值。本文论述了近年来对不溶性膳食纤维酶法改性所需的酶种类、方法及改性前后构成和生物作用效果的变化,为人们全面了解酶改性对膳食纤维的影响,为未来将酶改性的膳食纤维应用于食品提供理论基础,同时为可溶性膳食纤维工业化生产及其应用提供参考。     

Lipase-catalyzed direct and regioselective acylation of flavonoid glucoside for mechanistic investigation of stable plant pigments

One-step and regioselective acylation of flavonoid glucosides was achieved by lipase-catalyzed transesterification in dry organic media. For example, isoquercitrin (quercetin 3-O-glucoside), one of the flavonol monoglucosides, was converted efficiently to the corresponding aromatic acid ester (isoquercitrin 6'-O-cinnamate) by a lipase with vinyl cinnamate as an acyl donor. The method described, which is the first allowing the direct synthesis of aromatic acid esters of flavonoid glucosides, can be applied to the acylation of other glucosides, including naringin, rutin, callistephin, and chrysanthemin.     

脂溶性儿茶素类化合物的制备研究进展

儿茶素类化合物是一类具有多种生理活性的多酚类抗氧化剂,本文综述了国内外利用分子修饰法制备脂溶性儿茶素类化合物的进展。化学法主要利用酰氯在化学催化剂作用下反应制备脂溶性儿茶素,但是酰化反应发生的位点有所不同,总的来说酰基供体碳链长度一般大于等于8;酶法主要利用脂肪酶使脂肪酸或脂肪酸酯通过直接酯化或酯交换反应制备脂溶性儿茶素,在儿茶素及其化合物上键合烷基,由此提高了儿茶素类化合物在油脂体系中的溶解度,拓展了其应用领域。     

Effectiveness of rutin and its lipophilic ester in improving oxidative stability of sardine oil containing trace water

Poor oxidative stability exhibited by n‐3 polyunsaturated fatty acid rich sardine oil is a major challenge for its utilisation in industry. Considering the fact that water is always present in bulk oil in trace amounts during storage, an effort was made to understand and compare the effectiveness of rutin and its corresponding lipophilic ester in enhancing oxidative stability of refined sardine oil containing trace water (0.16% w/w). Peroxide value, conjugated diene value, p‐anisidine value and thiobarbituric acid reactive substances (TBARS) value were determined during 20 days storage. Rutin fatty ester showed 50% reduction in primary oxidation and 42.46% reduction in secondary oxidation, whereas rutin showed 20.6% and 20.43% reduction in primary and secondary oxidation, respectively, by the end of 20 days storage. Thus, it is clearly established that rutin fatty ester is more effective than hydrophilic rutin in sardine oil containing trace water, which contradicts the polar paradox theory.     

芦丁诱导猪肉肌原纤维蛋白结构变化对蛋白凝胶特性的改善作用

向猪肉肌原纤维蛋白氧化体系（40?mg/mL蛋白、10?μmol/L?FeCl3、100?μmol/L?VC和1?mmol/L?H2O2）中添加不同量的芦丁（0、10、50、100、150?μmol/g,以蛋白计）,测定蛋白质的巯基含量、表面疏水性、电泳、溶解度、凝胶强度和保水性、流变特性,研究芦丁对肌原纤维蛋白结构和凝胶特性的影响。结果表明,添加芦丁使肌原纤维蛋白的巯基含量显著下降（P＜0.05）,表面疏水性先下降后上升;芦丁使肌球蛋白重链（myosin heavy chain,MHC）强度减弱,肌动蛋白条带强度在其较高含量下（100?μmol/g和150?μmol/g）降低,添加β-巯基乙醇后,MHC和肌动蛋白条带大部分被还原;溶解度随着芦丁添加量的增加而降低;芦丁用量增加,蛋白的凝胶强度与保水性显著增强（P＜0.05）,凝胶最终形成阶段的储能模量（G’）提高。因此,芦丁可通过与肌原纤维蛋白的共价交联或适度增加蛋白的表面疏水性而改善蛋白的凝胶特性。