Enzymatically Modified Fats Based on Mutton Tallow and Rapeseed Oil Suitable for Fatty Emulsions

The aim of this work was to develop a new fat by enzymatic interesterification of mutton tallow with rapeseed oil. It was assumed that by inducing hydrolysis of fats by addition of water to the enzymatic preparation (8, 10, 15 wt%) natural emulsifiers would be produced in the reaction environment. Fat blends obtained from the enzymatic reactions were evaluated as a fat base for emulsion systems. It was found that the fat resulting from interesterification in the presence of Lipozyme RM IM (immobilized lipase from Rhizomucor miehei, Novozymes Bagsvaerd, Denmark) with 15 wt% of water possessed the highest content of polar fraction (MAG and DAG), and served as the most suitable blend for emulsification, producing emulsions that exhibited the highest stability.     

Characterization of Dispersion Systems Prepared with Mutton Tallow/Hemp Seed Oil-Based Diacylglycerols Using Ultrasonic or Mechanical Homogenization

The objective of this work was to compare the physical stability and physicochemical properties of emulsions, containing enzymatically modified fatty base and homogenized mechanically or by ultrasounds. In the study, lipase-catalyzed interesterification of mutton tallow and hemp seed oil, in a ratio of 3:1, 3:3, and 1:3 w/w, was performed in order to produce fatty bases of the emulsions. Reaction conditions were selected to obtain increased amount of the by-products (MAG and DAG), which were applied as the only emulsifiers in dispersion systems. The higher ratio of animal fat in the interesterified fatty basis of an emulsion had an impact on the greater stability of these systems. The correlation between thickening agent concentration in the prepared emulsions and stability was not observed. Smaller particle size was found in emulsions manufactured by ultrasonic homogenization, although it did not contribute to greater long-term stability of these emulsions. It was concluded that emulsions containing enzymatically modified fats and homogenized mechanically revealed greater physical stability than their counterparts homogenized with ultrasounds.     

Trans‐free fats through interesterification of canola oil/palm olein or fully hydrogenated soybean oil blends

Binary blends of canola oil (CO) and palm olein (POo) or fully hydrogenated soybean oil (FHSBO) were interesterified using commercial lipase, Lypozyme TL IM, or sodium methoxide. Free fatty acids (FFA) and soap content increased and peroxide value (PV) decreased after enzymatic or chemical interesterification. No difference was observed between the PV of enzymatically and chemically interesterified blends. Enzymatically interesterified fats contained higher FFA and lower soap content than chemically prepared fats. Slip melting point (SMP) and solid‐fat content (SFC) of CO and POo blends increased, whereas those of CO and FHSBO blends decreased after chemical or enzymatic interesterification. Enzymatically interesterified CO and POo blends had lower SMP and SFC (at some temperatures) than chemically interesterified blends. The status was reverse when comparing chemically and enzymatically interesterified CO and FHSBO blends. The induction period for oxidation at 120°C of blends decreased after interesterification. However, chemically interesterified blends were more oxidatively stable than enzymatically interesterified blends. Interesterified blends of CO and POo or FHSBO displayed characteristics suited to application as trans‐free soft tub, stick, roll‐in and baker's margarine, cake shortening and vanaspati fat.     

Enzymatic Production of Highly Unsaturated Monoacyglycerols and Diacylglycerols and Their Emulsifying Effects on the Storage Stability of a Palm Oil Based Shortening System

MDs [monoacylglycerols (MAGs) and diacylglycerols (DAGs) mixture] are widely‐used emulsifiers in specialty fats industrial production. An enzymatic production of highly unsaturated MDs (HUSMDs) and its effects on the storage stability of a palm oil‐based shortening system are reported. Oleic acid and corn oil were used to produce HUSMDs in a bubble column reactor (BCR) system in the presence of Novozyme 435. Under the optimized reaction conditions, the content of HUSMDs in the products was above 82 wt% with 46.67 wt% of MAGs and 35.56 wt% DAGs, respectively. Moreover, in the subsequent evaluation of MDs’ effects on the storage stabilities of a palm oil‐based shortening system (IEPO), HUSMDs proved to be a potent emulsifier with decent aeration properties and a possible alternative to saturated MAGs and DAGs (SMDs) made from fully hydrogenated high erucic acid colza oil. Compared with SMDs, HUSMDs decreased the crystallization rate significantly. The microstructure of them shows improved stability of β′ crystals, and no obvious aggregation of crystals was recorded in IEPO with HUSMDs, which also demonstrated the most stable hardness.     

Enzymatic interesterification of tallow-sunflower oil mixtures

In an effort to improve the physical and/or thermal characteristics of solid fats, the enzymatic interesterification of tallow and butterfat with high-oleic sunflower oil and soybean oil was investigated. The two simultaneously occurring reactions, interesterification and hydrolysis, were followed by high-performance liquid chromatography of altered glycerides and by gas-liquid chromatography of liberated free fatty acids. The enzymes used in these studies were immobilized lipases that included either a 1,3-acyl-selective lipase or acis-9-C₁₈-selective lipase. The degree of hydrolysis of the fat/oil mixtures was dependent upon the initial water content of the reaction medium. The extent of the interesterification reaction was dependent on the amount of enzyme employed but not on the reaction temperature over the range of 50–70°C. Changes in melting characteristics of the interesterified glyceride mixtures were followed by differential scanning calorimetry of the residual mixed glycerides after removal of free fatty acids. Interesterification of the glyceride mixes with the two types of enzymes allowed for either a decrease or increase in the solid fat content of the initial glyceride mix.     

Detection of lard and randomized lard as adulterants in refined-bleached-deodorized palm oil by differential scanning calorimetry

A study was conducted to assess the use of differential scanning calorimetry (DSC) for detecting the presence of lard/randomized lard as adulterants in refined-bleached-deodorized (RBD) palm oil. Lard extracted from the adipose tissues of pig was chemically interesterified using sodium methoxide as catalyst. DSC thermal profiles of both genuine lard and randomized lard were compared with those of other common animal fats such as beef tallow, mutton tallow, and chicken fat. Lard and randomized lard were then blended with RBD palm oil in two series, in proportions ranging from 0.2 to 20%, and DSC analyses were obtained. The DSC cooling profiles of adulterated RBD palm oil samples showed an adulteration peak corresponding to lard/randomized lard in the low-temperature region. This peak was confirmed as an indicator of the presence of lard in RBD palm oil since similar experiments carried out using other common animal fats such as mutton tallow, beef tallow, and chicken fat showed that the lard adulteration peak could be distinctly identified. Using this method, a detection limit of 1% lard/randomized lard was reached (P<0.0001).     

Lipase-catalyzed interesterification of oils and fats

Extracellular microbial lipases can be used as catalysts for the interesterification of oils and fats. Use of specific lipases gives products which are unobtainable by chemical interesterification methods. Some of these products have properties of value to the oils and fats industry. The catalysts for enzymatic interesterification are prepared by coating inorganic support materials with the lipases. For batch interesterification reactions, the catalyst particles are activated by addition of a small amount of water and then stirred with a reactant mixture dissolved in petroleum ether. At the end of the reaction period, the catalyst particles are removed by filtration, and the interesterified triglycerides isolated by conventional fat fractionation techniques. The catalyst can be used in subsequent batch reactions. As an alternative to the batch reaction system, continuous enzymatic interesterification processes can be operated by pumping water containg feedstock through a packed bed of activated catalyst.     

Storage stability of margarines produced from enzymatically interesterified fats compared to those prepared by conventional methods – Chemical properties

In this study, four margarine hardstocks were produced, two from enzymatically interesterified fats at 80 and 100% conversion, one from chemically randomized fat and one from physically mixed fat. These four hardstocks, blended with 50% sunflower oil, were mainly used for the production of table margarines in a pilot plant. Storage stability studies were carried out at storage temperatures of 5 and 25 °C for 12 wk. Margarines from the enzymatically interesterified fats were compared to the margarines produced by the conventional methods (chemical interesterification and physical blending) and to selected commercial margarines. The changes in the chemical properties of the products, including peroxide values (PV), tocopherols, free fatty acids, volatile oxidation products, and sensory evaluation, were examined during storage. It was observed that the margarine produced from the chemically interesterified fat had higher PV in weeks 4, 8 and 10 than the margarines produced from the enzymatically interesterified fats and the physically blended fat. These differences were not caused by different contents of tocopherols in the hardstocks. The differences between the processes for chemical and enzymatic interesterification, including further treatment stages, might be responsible for the development of a high PV in the margarine produced from the chemically interesterified fat. However, the contents of volatiles did not show the same tendency as observed for PV for the margarines stored at 25 °C during 12 wk. Storage at 25 °C accelerated oxidation compared to storage at 5 °C. The content of δ‐ and γ‐tocopherols decreased faster than the content of α‐ and β‐tocopherols during storage. This phenomenon was only affected by storage time, not by storage temperature. Sensory analysis did not show consistent differences between the produced margarines and commercial margarines, and no hydrolysis occurred for these four margarines during storage. The margarines produced from the enzymatically interesterified fats had low PV and a similar taste and smell compared to the margarine produced from the chemically interesterified fat.     

Characteristics of Eutectic Compositions of Restructured Palm Oil Olein,Palm Kernel Oil and Their Mixtures

The physico-chemical characteristics of blends of palm olein and palm kernel oil which were further modified by chemical interesterification were studied. The slip melting points of non-interesterified blends were 19.7, 16.2, 14.5, 14.5 and 14.4 °C while those of the chemically interesterified blends were 17.7, 16.2, 19.8, 18.7 and 18.7 °C at 40, 30, 20, 10 and 0% palm kernel oil, respectively. Chemical interesterification lowered the solid fat content of the pure samples and blends across different temperatures except 90% palm olein at 15 °C where the solid fat content was higher than for non-interesterified samples. Palm kernel oil, palm olein and their blends before and after chemical interesterification, crystallized mainly in the β′ form. However, chemical interesterification modified the microstructure from a combination of fat particles with void regions of crystalline materials to fat particles without regions of void crystalline materials. Palm olein and palm kernel oil blends are mainly used for food preparation in Nigeria. This study has shown that there are no significant differences in the physical and chemical properties of non-chemically interesterified and chemically interesterified blends of palm olein and palm kernel oil. This implies that blending of palm olein and palm kernel oil without chemical interesterification can provide the fluidity desirable at ambient temperatures for food applications in the tropics.     

Microstructure and Thermal Profile of Structured Lipids Produced by Continuous Enzymatic Interesterification

Enzymatic interesterification has been shown to be an alternative for the production of structured lipids resembling human milk fat. The knowledge of the physical properties of fat is an important tool for the implementation of this fat in a food matrix. The enzymatic interesterification reaction modifies the composition of triacylglycerols changing the crystallization properties and polymorphic form of fats. Blends containing different proportions of lard and soybean oil (80:20, 70:30, 40:60, 30:70 and 20:80) were enzymatically interesterified in a continuous flow tubular reactor and analyzed for crystalline structure by polarized light microscopy, the polymorphic form using X-ray diffraction and thermal properties by differential scanning calorimetry. The structural modifications resulting from continuous enzymatic interesterification changed the crystallization behavior and thermal profile of the samples, reducing enthalpy values. Structural changes were also evident on polarized light microscopy images, disclosing an increase in the crystallization rate among the samples after the continuous enzymatic reaction.     

Chemical and enzymatic interesterification of a beef tallow and rapeseed oil equal‐weight blend

A mixture of beef tallow and rapeseed oil (1:1, wt/wt) was interesterified using sodium methoxide or immobilized lipases from Rhizomucor miehei (Lipozyme IM) and Candida antarctica (Novozym 435) as catalysts. Chemical interesterifications were carried out at 60 and 90 °C for 0.5 and 1.5 h using 0.4, 0.6 and 1.0 wt‐% CH₃ONa. Enzymatic interesterifications were carried out at 60 °C for 8 h with Lipozyme IM or at 80 °C for 4 h with Novozym 435. The biocatalyst doses were kept constant (8 wt‐%), but the water content was varied from 2 to 10 wt‐%. The starting mixture and the interesterified products were separated by column chromatography into a pure triacylglycerol fraction and a nontriacylglycerol fraction, which contained free fatty acids, mono‐, and diacylglycerols. It was found that the concentration of free fatty acids and partial acylglycerols increased after interesterification. The slip melting points and solid fat contents of the triacylglycerol fractions isolated from interesterified fats were lower compared with the nonesterified blends. The sn‐2 and sn‐1,3 distribution of fatty acids in the TAG fractions before and after interesterification were determined. These distributions were random after chemical interesterification and near random when Novozym 435 was used. When Lipozyme IM was used, the fatty acid composition at the sn‐2 position remained practically unchanged, compared with the starting blend. The interesterified fats and isolated triacylglycerols had reduced oxidative stabilities, as assessed by Rancimat induction times. Addition of 0.02% BHA and BHT to the interesterified fats improved their stabilities.     

Functions of emulsifiers in food systems

The function of food grade emulsifiers in various food products (emulsions, starch based food, yeast raised bakery products, etc.) are reviewed. The stability of emulsions against coalescence of dispersed droplets is among other factors dependent on monoor multimolecular interfacial films with viscoelastic properties formed by adsorbed emulsifier molecules. Agglomeration of fat globules in whippable emulsion is needed to obtain desired foam stability and texture and can be controlled by lipophilic emulsifiers. Complex formation with starch components (amylose) is influenced both by the chemical structure of an emulsifier and by its physical behaviour in water. Interaction with proteins takes place primarily with anionic emulsifiers or very hydrophilic, nonionic types, which thereby improves the rheological properties of wheat gluten. Emulsifiers are also used as crystal modifying agents in fats where polymorphic changes during storage creates texture problems.     

Application of palm olein in the production of zero‐trans Iranian vanaspati through enzymatic interesterification

The utilization of palm olein in the production of zero‐trans Iranian vanaspati through enzymatic interesterification was studied. Vanaspati fat was made from ternary blends of palm olein (POL), low‐erucic acid rapeseed oil (RSO) and sunflower oil (SFO) through direct interesterification of the blends or by blending interesterified POL with RSO and SFO. The slip melting point (SMP), the solid fat content (SFC) at 10–40 °C, the carbon number (CN) triacylglycerol (TAG) composition, the induction period (IP) of oxidation at 120 °C (IP₁₂₀) and the IP of crystallization at 20 °C of the final products and non‐interesterified blends were evaluated. Results indicated that all the final products had higher SMP, SFC, IP of crystallization and CN 48 TAG (trisaturated TAG), and lower IP₁₂₀, than their non‐interesterified blends. However, SMP, SFC, IP₁₂₀, IP of crystallization and CN 48 TAG were higher for fats prepared by blending interesterified POL with RSO and SFO. A comparison between the SFC at 20–30 °C of the final products and those of a commercial low‐trans Iranian vanaspati showed that the least saturated fatty acid content necessary to achieve a zero‐trans fat suitable for use as Iranian vanaspati was 37.2% for directly interesterified blends and 28.8% for fats prepared by blending interesterified POL with liquid oils.     

Physical and chemical properties of randomly interesterified blends of soybean oil and tallow for use as margarine oils

Refined, unhydrogenated soybean oil and edible beef tallow were interesterified with sodium methoxide. This was done as an alterna-tive to hydrogenation for the production of plastic fats for use as margarine oils. Using 0.5% sodium methoxide at 80 C, interesterifi-cation was complete in 30 min as determined by lipase hydrolysis. A blend of 60% soybean oil and 40% edible beef tallow was found to have physical characteristics (melting point, solid fat index) similar to those of commercial tub margarine oils. The level of poly-unsaturated fatty acids was slightly lower and the level of saturated fatty acids slightly higher than the commercial margarine oils. Iodine value andtrans fatty acid determinations indicated no dis-cernible effect on the degree of unsaturation or the level of isomeric fatty acids by the interesterification process. The interesterified blend did contain 3.0%trans fatty acids which were originally present in the tallow. Oxidative stability of the interesterified oils was estimated by peroxide value determinations over several days on samples stored at 60 C. Experimental blends treated with 0.1% citric acid had poorer stability than the partially hydrogenated margarine oils; however, 0.01% BHA significantly delayed oxidation of the experimental samples. Presented at the JOCS/AOCS annual meeting, 1979, San Francisco. Paper No. 6797, Journal Series, Nebraska Agricultural Experiment Station.     

Solidification and phase transformation behaviour of food fats — a review

Recent studies on physical behaviour of food fats have been reviewed, with an emphasis on solidification properties in bulk and emulsion states. It was shown that the use of synchrotron radiation X-ray beam highlighted the solidification behaviour of polymorphic fats, which was not unveiled by traditional X-ray beams on a laboratory scale as summarised in the following: (a) kinetic processes of molecular ordering of lamella-structured fat crystals, (b) mixing behaviour of different triacylglycerols forming miscible, eutectic, and molecular compound crystals, and (c) remarkable influences of shearing forces on the rapid solidification of the more stable forms of cocoa butter. These results have given some indications to the physical control of fat solidification, and the blending and fractionation of solid fats. The influences of hydrophobic emulsifiers on the solidification of fat crystals in O/W emulsions, which were in-situ monitored by an ultrasound velocity technique, have shown that the addition of highly hydrophobic emulsifiers having long-saturated acid moieties accelerated the nucleation of the fats at the interface areas of the emulsions. These effects are also indicative for the selection of the emulsifier for the control of the fat solidification in the emulsion phases.     

Enzymatic interesterification of palm stearin and palm kernel olein

Palm stearin (POs) and palm kernel olein (PKOo) blends were modified by enzymatic interesterification (IE) to achieve the physical properties of margarine fats. POs and PKOo are both products of the palm oil industry that presently have limited use. Rhizomucor miehei lipase (Lipozyme IM 60) was used to catalyze the interesterification of oil blends at 60°C. The progress of interesterification was monitored by following changes in triacylglyceride composition. At 60°C interesterification can be completed in 5 h. Degrees of hydrolysis obtained through IE for all blends were decreased from 2.9 to 2.0 by use of dry molecular sieves. The solid fat contents of POs/PKOo 30:70 and 70:30 interesterified blends were 9.6 and 18.1 at 20°C, and 0 and 4.1 at 35°C, respectively. The slip melting point (SMP) of POs/PKOo 30:70 was 40.0°C before interesterification and 29.9°C after IE. For POs/PKOs 70:30, SMP was 47.7 before and 37.5°C after IE. These thermal characteristics of interesterified POs/PKOo blend ratios from 30:70 to 70:30 were comparable to those of commercial margarines. Results showed that IE was effective in producing solid fats with less than 0.5% trans.     

Stability of Interesterified Fats

Stability study carried out with interesterified (i) refined cottonseed oil (CSO), blends of (ii) vanaspati and refined groundnut oil (40:60 w/w) (vanaspati-GNO), (iii) refined and bleached sal (Shorea robusta) fat and refined groundnut oil (30:70) (Sal-GNO) and (iv) edible grade sheep tallow and refined groundnut oil (30:70) (ST-GNO) indicated that rearranged CSO and ST-GNO were more and sal-GNO less prone to oxidative rancidity compared to their corresponding starting stocks. Vanaspati-GNO was more or less similar to its starting stock. In other words, the process of interesterification had different effects on different starting stocks vis-à-vis oxidative rancidity. Addition of antioxidant (BHA) improved the stability of interesterified products to a large extent. Development of oxidative rancidity as measured by peroxide value in all the interesterified products during storage was faster than that observed with vanaspati. All the interesterified products were stable towards hydrolytic rancidity during storage. When the linoleic content of a blend was about 25%, the process of interesterification had no or marginal adverse effect on the development of oxidative rancidity. Higher linoleic acid (about 50%) made the stock more unstable as a result of interesterification. Comparatively more instability of interesterified blend containing sheep tallow cannot be explained on the basis of its linoleic acid content and its migration as a result of rearrangement.     

Interesterification,chemical or enzymatic catalysis

The interesterification process is one of the oil modification processes the refiner can use to alter the physical properties of oils and fats, The reaction requires a catalyst to proceed. This can be a base or a lipase enzyme. In the currently accepted mechanism of the base‐catalysed interesterification reaction, two anionic intermediates are involved: the enolate anion and the glycerolate anion. The presence of the enolate anion explains why an equivalent amount of FAMEs are formed when sodium methanolate is added to oil and why FFAs are formed when the catalysts is inactivated with water. Based on this insight, process development can aim at avoiding these by‐products and thereby increase the cost advantage of the chemical process over the enzymatic process even further. The chemical process is also more flexible than the solely continuous enzymatic process, which latter requires extensive purification of the oil to be interesterified.     

Changes in the Contents of Cholesterol and Cholesteryl Esters Occurring during Lipase-Catalysed Interesterification Reactions

The free cholesterol and cholesteryl esters isolated on silica column and the total cholesterol from saponified fat samples obtained from untreated butter fat and butter fats interesterified with Pseudomonas fluorescens lipase as catalyst were determined on immobilised OV-351 and phenylmethylsilicone gas chromatographic columns. Fatty acid composition of the cholesteryl esters of the untreated butter fat differed markedly from the fatty acid composition of acylglycerols. The proportions of cholesteryl palmitate, stearate and oleate were clearly higher in the interesterified fats and the proportion cholesteryl linoleate was distinctly lower. Determination of the free cholesterol and cholesteryl esters and the total cholesterol in the fraction of unsaponifiables revealed that, on average, 89% of cholesterol is esterified in the interesterification products, but only 2% in intact butter fat.     

Physicochemical Properties of Interesterified Blends of Fully Hydrogenated Crambe abyssinica Oil and Soybean Oil

The chemical interesterification of blends of soybean (SO) and fully hydrogenated crambe oil (FHCO) in the ratios of 80:20, 75:25, 70:30, 65:35, and 60:40 (w/w), respectively, was investigated. FHCO is a source of behenic acid. The blends and the interesterified fats were analyzed for fatty acid and triacylglycerol composition, regiospecific distribution, slip melting point, solid profile, and consistency. The regiospecific analysis of the TAG indicated random insertion of saturated fatty acids at sn-2 of the glycerol of the interesterified blends with more significant alterations at sn-2 than at sn-1 and sn-3. The gradual addition of FHCO increased the solid fat content and the slip melting point. The chemical interesterification formed new TAG facilitating the miscibility between SO and FHCO. The 70:30 interesterified blend was suitable for general use, 60:40 for use as a base stock. At 35 °C, the 65:35 interesterified blend showed suitable plasticity for use in products with fat contents below 80 %. FHCO, rich in behenic acid, is not associated with increased total cholesterol and LDL cholesterol, and it can be used as a low trans fat. FHCO is not associated with increased total cholesterol and LDL cholesterol, and it can be used as a low trans fat alternative.