Utilizing biotechnology in producing fats and oils with various nutritional properties

The role of dietary fat in health and wellness continues to evolve. In today's environment, trans fatty acids and obesity are issues that are impacted by dietary fat. In response to new information in these areas, changes in the amount and composition of edible fats and oils have occurred and are occurring. These compositional changes include variation in fatty acid composition and innovation in fat structure. Soybean, canola, and sunflower are examples of oilseeds with varied fatty acid composition, including mid-oleic, high-oleic, and low-linolenic traits. These trait-enhanced oils are aimed to displace partially hydrogenated vegetable oils primarily in frying applications. Examples of oils with innovation in fat structure include enzyme interesterified (EIE) fats and oils and diacylglycerol oil. EIE fats are a commercial edible fat innovation, where a lipase is used to modify the fat structure of a blend of hard fat and liquid oil. EIE fats are aimed to displace partially hydrogenated vegetable oils in baking and spread applications. Diacylglycerol and medium-chain triglyceride (MCT)-based oils are commercial edible oil innovations. Diacylglycerol and MCT-based oils are aimed for individuals looking to store less of these fats as body fat when they are used in place of traditional cooking and salad oils.     

Lipase incorporated ionic liquid polymers as active, stable and reusable biocatalysts

The aim of this study was to develop ionic liquid (IL) polymer materials incorporating enzymes that can be used as active, stable and reusable biocatalysts. To this goal, Candida rugosa lipase has been microencapsulated in surfactant aggregates formed in an IL monomer or the solution of an IL monomer/IL and then incorporated into polymer frameworks through the free radical polymerization of an IL (1-vinyl-3-ethylimidazolium bis(trifluoromethyl-sulfonyl) amide) ([veim][Tf(2)N]). The activity, stability and reusability of such IL polymer materials containing lipase were evaluated using lipase-catalyzed hydrolysis of p-nitrophenyl butyrate (p-PNB) as a model reaction. Lipase encapsulated within ionic liquid polymer materials remained active and exhibited excellent stability in aqueous solutions. More importantly, these biopolymer materials retained most of their activity after five reaction cycles, in which biopolymers were recovered from the reaction mixture simply by centrifugation. This study promulgates a direction toward the design of IL - an interesting class of tunable and designable solvents - based polymer materials containing biomolecules via a combination of polymer and supramolecular chemistry for diverse applications.     

Increased enantioselectivity and remarkable acceleration of lipase-catalyzed transesterification by using an imidazolium PEG-alkyl sulfate ionic liquid

Several types of imidazolium salt ionic liquids were prepared derived from poly(oxyethylene)alkyl sulfate and used as an additive or coating material for lipase-catalyzed transesterification in an organic solvent. A remarkably increased enantioselectivity was obtained when the salt was added at 3-10 mol % versus substrate in the Burkholderia cepacia lipase (lipase PS-C)-catalyzed transesterification of 1-phenylethanol by using vinyl acetate in diisopropyl ether or a hexane solvent system. In particular, a remarkable acceleration was accomplished by the ionic liquid coating with lipase PS in an iPr(2)O solvent system while maintaining excellent enantioselectivity; it reached approximately 500- to 1000-fold acceleration for some substrates with excellent enantioselectivity. A similar acceleration was also observed for IL 1-coated Candida rugosa lipase. MALDI-TOF mass spectrometry experiments of the ionic-liquid-coated lipase PS suggest that ionic liquid binds with lipase protein.     

An improved method for sn-2 position analysis of triacylglycerols in edible oils and fats based on immobilised lipase D (Rhizopus delemar)

The use of lipase D (Rhizopus delemar) immobilised on microporous polypropylene as a replacement for the standard pancreatic lipases used in the stereospecific sn-2 position analysis of triacylglycerols from edible oils and fats is studied. Excellent hydrolysis characteristics are obtained in hexane/methanol solvents at reaction temperatures up to 60 degrees C with hydrolysis times of only 10-20 min. The favourable conditions for the hydrolysis reaction allow fats with higher melting points to be analysed and facilitate coupling of the hydrolysis reaction to the later steps in the analytical protocol. The performance of the new method is compared to that of the standard method using pancreatic lipase. The novel procedure is faster, manual sample handling is reduced, while the results obtained with both methods are comparable. The influence of alkyl-chain length on hydrolysis rates seems to be negligible for the most common vegetable fatty acids. Acyl migration was found to be absent. The short-term repeatability of the method ranges from 10% for fatty acids present at levels close to the detection limits to less than 1% for the major fatty acids. The detection limit is approximately 0.05%. Although the application of the immobilised enzyme in fully automated sn-2 position analysis seems to be promising, the attempts to do this using a packed bed reactor were not successful due to a rapid loss of enzyme activity.     

Dicationic ionic liquid stationary phase for GC-MS analysis of volatile compounds in herbal plants

The seeming “dual nature” of ionic liquids (ILs) for separating both apolar and polar compounds suggests that ILs may have a great potential for complex samples like essential oils from herbal plants that contain a great variety of compounds. In the present work, a geminal dicationic IL, 1,9-di(3-vinylimidazolium)nonane bis[(trifluoromethyl)sulfonyl]imidate, was investigated for this purpose. To find the best way to achieve satisfactory separations simultaneously for the compounds in essential oils, the dicationic IL was used as the stationary phase for capillary gas chromatography (GC) in two ways, either in its pure state or as a mixed stationary phase with monocationic ILs and a polysiloxane diluent. Interestingly, it was found that the mixed stationary phase exhibited a much better selectivity for polar and nonpolar compounds than either the dicationic IL or the polysiloxane, suggesting that a kind of synergistic effect occurred when these stationary phases were combined in the way described. A comparison with two commercial stationary phases (polar and nonpolar) indicated that this novel mixed stationary phase behaved in a way closer to a polar stationary phase in terms of selectivity and elution order. The present work demonstrates that the mixed stationary phase is efficient and selective and can be an alternative choice for the GC analysis of samples of complex composition. Figure Divinyldiimidazolium-based ionic liquid stationary phase     

An efficient fluorination of β-ketosulfones promoted by a room-temperature ionic liquid at ambient conditions under ultrasound irradiation using Selectfluor~(TM) F-TEDA-BF_4

<正>The fluorination reaction involving aβ-ketosulfones by Selectfluor~(TM) was efficiently promoted by the ionic liquid,[Hbim]BF_4 (EL) as a reaction medium with methanol as co-solvent at room temperature under ultrasonic irradiation to afford the corresponding mono and difluoro-β-ketosulfones in excellent yields.The advantages of this method include among others the use of a recyclable, non-volatile ionic liquid,which promotes this protocol under room temperature without the requirement of any added catalyst under ultrasonic irradiation.     

水饱和离子液体中萘普生的酶法拆分

 考察了五种水饱和离子液体反应介质中固定化脂肪酶催化外消旋萘普生甲酯的水解. 结果表明,这些具有不同阴阳离子组合的咪唑基离子液体作为反应介质都能促进水解反应的有效进行,但在水饱和1-正丁基-3-甲基咪唑六氟磷酸盐(［bmim］PF6)中,反应进行得更快,转化率更高. 这些离子液体对脂肪酶均有一定的溶解作用并使溶解的脂肪酶失活. 脂肪酶在离子液体中的溶解度与其剩余活性之间有一定的相关性. 为了解决脂肪酶在离子液体中的溶解问题,使用非极性、高比表面积的无定形多孔硅胶YWG-C6H5对脂肪酶进行了固定化. 在水饱和［bmim］PF6中使用固定化脂肪酶催化反应,反应72 h的转化率为28.3%, 产物的对映体过量值为98.2%, 继续进行反应,转化率将增加,但产物的对映体过量值明显下降. 利用离子液体有别于传统有机溶剂的特性,对离子液体的循环使用、产物的回收和水的补充方法进行了研究. 在反复批式反应中,固定化脂肪酶连续使用五次,活性仅略微下降.     

Isolation, Characterization, and Catalytic Properties of a Novel Lipase Which Is Activated in Ionic Liquids and Organic Solvents

A novel extracellular lipase with organic solvent tolerance was isolated from a local Pseudomonas species. The lipase gene was cloned and expressed in Escherichia coli as a heterologous host and purified by affinity chromatography. The activity of purified lipase was investigated in the presence of imidazolium-based ionic liquids (ILs) such as EMIM[Cl], BMIM[Cl], and HMIM[Cl]. It has been found that the activity of treated lipase with ILs was higher than untreated control in the hydrolysis reaction. Also, the results indicated that the enzymatic activity strongly depends on IL concentration in reaction media. The best concentration of the IL was 30%, 45%, and 50% (v/v) for HMIM[Cl], BMIM[Cl], and EMIM[Cl], respectively. Additionally, the enzyme exhibited excellent stability in the presence of 25% of n-hexane, toluene, acetone, and t-butanol. The optimum values of pH and temperature were determined 10 and 55 °C, respectively. The K (m) and V (max) values were calculated 0.4 mM and 1.92 U/ml, respectively, using p-nitrophenyl palmitate as substrate. With respect to the biochemical properties of the newly isolated lipase such as high-level stability and noticeable activity in the presence of organic solvents and ionic liquids, the newly isolated lipase seems to be a good candidate for environmental and industrial processes carried out in non-aqueous media.     

MODIFICATION OF FATS AND OILS BY LIPASES

Enzymatic modification of fats and oils using lipases has recently come into limelight, giving rise to a breakthrough in modification technology. This paper describes the latest development and future trend of this new technology with respect to the enzyme lipases, lipase reactions, and bioreactor systems. Particular attention is paid to the utilization of this technology in industrial production of functional fats and oils. A new topic of the reasearch work of an enzymatic reaction of conversion from diglycerides to triglycerides in crude palm olein is presented in detail.     

Enzymatic conversion of waste cooking oils into alternative fuel—Biodiesel

Production of biodiesel from pure oils through chemical conversion may not be applicable to waste oils/fats. Therefore, enzymatic conversion using immobilized lipase based on Rhizopus orzyae is considered in this article. This article studies this technological process, focusing on optimization of several process parameters, including the molar ratio of methanol to waste oils, biocatalyst load, and adding method, reaction temperature, and water content. The results indicate that methanol/oils ratio of 4, immobilized lipase/oils of 30 wt% and 40°C are suitable for waste oils under 1 atm. The irreversible inactivation of the lipase is presumed, and a stepwise addition of methanol to reduce inactivation of immobilized lipases is proposed. Under the optimum conditions the yield of methyl esters is around 88–90%.     

柱状假丝酵母脂肪酶同工酶的一种高效液相色谱分离新方法

建立一种“疏水界面亲和色谱”分离柱状假丝酵母脂肪酶同工酶的高效液相色谱新方法。将商品化的CRL经离子交换色谱分离为两个同工酶组分 (CRLA和CRLB) ,在极低离子强度下 ,根据同工酶活性中心周围处于“开放”构象的疏水腔具亲疏水界面的特性 ,用疏水界面亲和色谱在NucleosilC4 (10 μ ,3 0 0 ,2 5 0× 4.60mm)柱上将CRLA和CRLB都分离为 4种同工酶组分。疏水界面亲和色谱非常适用于分离这种结构差异轻微的同工酶组分     

The [BMI][Tf2N] ionic liquid/water binary system: a molecular dynamics study of phase separation and of the liquid-liquid interface

We report molecular dynamics (MD) simulations of the aqueous interface of the hydrophobic [BMI][Tf2N] ionic liquid (IL), composed of 1-butyl-3-methylimidazolium cations (BMI+) and bis(trifluoromethylsulfonyl)imide anions (Tf2N-). The questions of water/IL phase separation and properties of the neat interface are addressed, comparing different liquid models (TIP3P vs TIP5P water and +1.0/-1.0 vs +0.9/-0.9 charged IL ions), the Ewald vs the reaction field treatments of the long range electrostatics, and different starting conditions. With the different models, the "randomly" mixed liquids separate much more slowly (in 20 to 40 ns) than classical water-oil mixtures do (typically, in less than 1 ns), finally leading to distinct nanoscopic phases separated by an interface, as in simulations which started with a preformed interface, but the IL phase is more humid. The final state of water in the IL thus depends on the protocol and relates to IL heterogeneities and viscosity. Water mainly fluctuates in hydrophilic basins (rich in O(Tf2N) and aromatic CH(BMI) groups), separated by more hydrophobic domains (rich in CF3(Tf2N) and alkyl(BMI) groups), in the form of monomers and dimers in the weakly humid IL phase, and as higher aggregates when the IL phase is more humid. There is more water in the IL than IL in water, to different extents, depending on the model. The interface is sharper and narrower (approximately 10 A) than with the less hydrophobic [BMI][PF6] IL and is overall neutral, with isotropically oriented molecules, as in the bulk phases. The results allow us to better understand the analogies and differences of aqueous interfaces with hydrophobic (but hygroscopic) ILs, compared to classical organic liquids.     

The heterogeneous catalytic heck reaction in an ionic liquid

The C-C cross coupling reaction between bromobenzene and styrene in the presence of Pd deposited on mesoporous acetylene soot (Pd/C) occurred in a tetraalkylammonium ionic liquid (IL) as a truly heterogeneous catalytic process. The active palladium form was not transferred into the reaction solution. Deposited palladium activation was observed when the Pd/C catalyst was preliminarily heated in the IL in the presence of dibutylamine as a base.     

Design and exploration of caffeine-based Brönsted acidic ionic liquid (CaffBAIL) for the synthesis of DHPMs,xanthenediones, and acridinediones

A novel caffeine-based acidic ionic liquid was synthesized, characterized, and utilized as an efficient and green catalyst for synthesizing DHPMs, xanthenediones, and acridinediones under solvent-free conditions. The prepared IL showed notable catalytic activity in organic reactions under solvent-free conditions with good reusability of four runs. The advantage of the present protocol is underlined by the originality of the prepared ionic liquid, absence of toxic solvents, high yields of products, and short reaction times.

     

Optimization of distilled monoglycerides production

Monoglycerides (MG) are emulsifiers widely used in food and pharmaceutical industries. Current industrial processes for MG production consist of the interesterification of triglycerides with glycerol (GL), in the presence of inorganic catalysts at high temperatures (>200 degrees C). This reaction is known as glycerolysis and produces a mixture of approx 50% of MG. This level of concentration is suitable for many applications, although, for some specific uses like margarine, shortening, icing, and cream filling, require distilled MGs, which are purified MG (min. 90%) obtained by the molecular distillation process. Therefore, in this work, a 2(3) factorial design was employed to evaluate the effects of reaction parameters in the MG content after the interesterification reaction of refined soybean oil with GL in the presence of sodium hydroxide as catalyst. After that, the MG content in the reaction product was enhanced through the molecular distillation process in order to obtain distilled MG.     

Ordered Mesoporous Organosilica with Ionic‐Liquid Framework: An Efficient and Reusable Support for the Palladium‐Catalyzed Suzuki–Miyaura Coupling Reaction in Water

The preparation of a novel palladium‐supported periodic mesoporous organosilica based on alkylimidazolium ionic liquid (Pd@PMO‐IL) in which imidazoilium ionic liquid is uniformly distributed in the silica mesoporous framework is described. Both Pd@PMO‐IL and the parent PMO‐IL were characterized by N₂‐adsorption–desorption, diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS), TEM, and solid‐state NMR spectroscopy. We have demonstrated that Pd@PMO‐IL is an efficient and reusable catalyst for the Suzuki–Miyaura coupling reaction of various types of iodo‐, bromo‐, and even deactivated aryl chlorides in water. It was also found that although the PMO‐IL nanostructure acts as reservoir for soluble Pd species, it can also operate as a nanoscaffold to recapture the Pd nanoparticles into the mesochannels thus preventing extensive agglomeration of Pd. This observation might be attributed to the isolated ionic liquid units that effectively control the reaction mechanism by preventing Pd agglomeration and releasing and recapturing Pd nanoparticles during the reaction process. The catalyst can be recovered and reused for at least four reaction cycles without significant loss of activity.     

己内酰胺对甲基苯磺酸离子液体中甲苯的选择性硝化反应

制备了一种低毒性的新型Brφnsted酸性离子液体己内酰胺对甲基苯磺酸([CP]PTSA)作为催化剂和溶剂,用于催化甲苯与等摩尔的体积分数为67%HNO3的硝化反应,用核磁共振、红外光谱对[CP]PTSA的结构进行了表征。考察了不同的反应温度、反应时间及催化剂用量等因素对甲苯硝化反应的产率和对位选择性的影响。得出最佳反应条件为:温度55℃,时间24h,离子液体用量占甲苯摩尔分数的20%,产物硝基甲苯中邻与对位异构体质量比为1.2,较硝-硫混酸硝化的相应比值(1.6)显著降低,硝基甲苯的产率达37.1%。离子液体重复使用4次仍表现出较好的催化活性及对位选择性。用质谱分析对回收的离子液体的结构进行了表征。结果表明,该离子液体的结构稳定。     

Lipase-catalyzed glucose fatty acid ester synthesis in ionic liquids

[reaction: see text] Glucose fatty acid ester synthesis with poly(ethylene glycol)-modified Candida antarctica lipase B (CAL-B) was performed in pure 1-butyl-3-methyl imidazolium tetrafluoroborate [BMIM][BF(4)] (30% conversion) and in pure 1-butyl-3-methyl imidazolium hexafluorophosphate [BMIM][PF(6)] (35% conversion). In a solvent system composed of ionic liquid and 40% t-BuOH conversions up to 90% and isolated yields of up to 89% were achieved using fatty acid vinyl esters as acyl donors and commercial CAL-B.     

A novel synthesis of 1-aryl tetrazoles promoted by employing the synergy of the combined use of DMSO and an ionic liquid as the solvent system at ambient temperature

The synergy of the combined use of DMSO and an ionic liquid viz. (bbim)⁺Br⁻ has brought about a mild, convenient, efficient, and rapid protocol for the synthesis of 1-substituted-1H-1,2,3,4-tetrazoles via the condensation of amines, triethyl orthoformate, and sodium azide at ambient temperature in excellent isolated yields (85-90%). The inherent Bronsted acidity of ionic liquid and high polarity of both IL and DMSO resulted in a significant enhancement in the reaction rate.     

Optimization of distilled monoglycerides production

Monoglycerides (MG) are emulsifiers widely used in food and pharmaceutical industries. Current industrial processes for MG production consist of the interesterification of triglycerides with glycerol (GL), in the presence of inorganic catalysts at high temperatures (>200°C). This reaction is known as glycerolysis and produces a mixture of approx 50% of MG. This level of concentration is suitable for many applications, although, for some specific uses like margarine, shortening, icing, and cream filling, require distilled MGs, which are purified MG (min. 90%) obtained by the molecular distillation process. Therefore, in this work, a 2³ factorial design was employed to evaluate the effects of reaction parameters in the MG content after the interesterification reaction of refined soybean oil with GL in the presence of sodium hydroxide as catalyst. After that, the MG content in the reaction product was enhanced through the molecular distillation process in order to obtain distilled MG.