Interesterification Research Articles

Enzymatic Interesterification of Crude Palm Oil with Methyl acetate: Effect of Pre-treatment, Enzyme’s Dosage and Stability

In the present study, biodiesel was produced via the enzymatic interesterification of Crude Palm Oil (CPO) and methyl acetate within ultrasonic condition. In contrast to alcohol, methyl acetate as an acyl acceptor does not inhibit lipase activity and can create triacetin as a useful byproduct. In this work, Immobilized lipase from Candida Antartica A (CaLA) was utilized as biocatalyst and the effect of using non-pretreated CPO and pre-treated CPO as feedstock were explored. The pre-treatment of CPO involves degumming with acid, washing with water, and bleaching. The enzymatic interesterification was conducted in three-neck flasks using an ultrasonic water bath at 45o C. Few parameter effects on biodiesel production were also investigated, including the effect of molar ratio of CPO to methyl acetate, the effect amount of lipase, and the reusability of immobilized lipase (CaLA) in the interesterification reaction. The highest average Biodiesel yield of 80.6% was obtained from pretreated CPO at a molar ratio of 1:9 with 100 mg (1% w/v) of Immobilized CaLA, after three hours of reaction. Further research on the reusability of immobilized CaLA revealed that the yield of biodiesel reduced significantly after the second run. The results of the present study also demonstrated that Immobilized CaLA performed well at low concentrations but had low stability, with productivity decreasing to 92% upon reuse after the initial run. In order to make Immobilized lipase economically viable, further research must be conducted to overcome its low stability in the reaction.

Enzymatic interesterification improves the lipid composition, physicochemical properties and rheological behavior of Cinnamomum camphora seed kernel oil, Pangasius bocourti stearin and perilla seed oil blends

The study aimed to investigate the effect of enzymatic interesterification on the lipid composition, physicochemical properties and rheological behavior of Cinnamomum camphora seed kernel oil (CCSKO), Pangasius bocourti stearin (PBST) and perilla seed oil (PSO) blends. The results showed that the interesterification process significantly changed the TAG profile of the blends. Lipid products from the enzymatic interesterification (EIE) had significantly lower slide melting point and solid fat content than the non-interesterification (NIE) lipid products. Interesterification process changed the crystal polymorphic forms from β > β′ of NIE to β < β′ of EIE. The crystal morphology of EIE was smaller and more diffuse compared to the NIE. Moreover, EIE showed improved rheological behavior, which was more suitable for food margarine preparation. The findings have provided a theoretical basis for the potential application of Lipozyme TL IM modified lipid products in the food industry.

The effect of enzymatic interesterification on the high oleic-high stearic sunflower oil fractionation and the physico-chemical properties of stearins

Oil fractionation processes typically involve the crystallization of the most saturated triacylglycerols to facilitate the separation of the liquid phase. However, the fractionation of new high stearic oils derived from sunflower (HOHS) is not as efficient as that of tropical fats. In the present work we studied the effect of enzymatic inter esterification (EIE) on the HOHS sunflower oil fractionation. EIE altered the properties and melting point of the HOHS oil, so it eased fractionation allowing operation at lower temperatures (8 °C). These fractionations produced stearins with high saturated fatty acid contents at higher yields than the non-EIE oil (28.3% vs 18.7%). The melting profile of the stearins were also characterized. EIE stearins displayed broader melting profiles with higher crystallization onsets (from 26.3 °C to 34 °C). Fractionations proved to be reproducible at pilot plant scale. The resulting stearins exhibited a solid content of 30% at 20 °C and displayed solidification curves similar to palm-based filling fats. The results indicate that changes in fatty acid distribution in the TAG backbone critically affect TAG crystallization, which was faster and more reproducible, not requiring prior dewaxing and seeding. Moreover, the fractionation products differed, yielding stearins with higher levels of solids.

Physicochemical and technological properties of pracaxi oil, cupuassu fat and palm stearin blends enzymatically interesterified for food applications

Pure, blends and enzymatically interesterified blends in different proportions of pracaxi oil/cupuassu fat (PC) and pracaxi oil/palm stearin (PS) (w/w) (40:60, 50:50, 60:40 and 70:30), were characterized. Samples with the greatest food interest (70:30 w/w PC and PS) were characterized by X-ray diffraction, optical microscopy, and DSC. β crystalline form was reduced in interesterified blends. Crystals in the blends showed stable polymorphic forms, whereas interesterified blends showed a reduction in β crystalline form, and crystals in α form. PC and PS blends with pracaxi oil (70:30 w/w) showed the best atherogenicity (AI) 0.21 and 0.34 and thrombogenicity (TI) 0.38 and 0.55 indexes, respectively. Pracaxi oil has an oxidative stability of 2.32 h. Fats (cupuassu and palm stearin) has stability (3.60–6.51 h), and contributed to increase the oxidative stability of the blends. Pracaxi oil and cupuassu fat (40:60) reached oxidative stability of 3.85 h, while with palm stearin in the same proportion the stability increased to 4.34 h. Enzymatic interesterification increased the plastic range (SFC at temperatures up to 35 °C) of PC blends and reduced the SFC of PS blends. Interesterification caused a reduction in the consistency of the blends (p < 0.05), which shows the improvement in textural properties.

Trends in Fat Modifications Enabling Alternative Partially Hydrogenated Fat Products Proposed for Advanced Application

The natural properties of oils and fats do not always allow for their direct use in industry (e.g., for food, cosmetics, and pharmaceuticals). Furthermore, such raw materials are often too expensive. Nowadays, the requirements for the quality and safety of fat products are increasing. For this reason, oils and fats are subjected to various modifications that make it possible to obtain a product with the desired characteristics and good quality that meets the needs of product buyers and technologists. The modification techniques of oils and fats change their physical (e.g., raise the melting point) and chemical properties (e.g., fatty acid composition). Conventional fat modification methods (hydrogenation, fractionation, and chemical interesterification) do not always meet the expectations of consumers, nutritionists, and technologists. In particular, Hydrogenation, while it allows us to obtain delicious products from the point of view of technology, is criticised for nutritional reasons. During the partial hydrogenation process, trans-isomers (TFA), dangerous for health, are formed. One of the modifications that meets current environmental requirements and trends in product safety and sustainable production is the enzymatic interesterification of fats. The unquestionable advantages of this process are the wide spectrum of possibilities for designing the product and its functional properties. After the interesterification process, the biologically active fatty acids in the fatty raw materials remain intact. However, this method is associated with high production costs. Oleogelation is a novel method of structuring liquid oils with small oil-gelling substances (even 1%). Based on the type of oleogelator, the methods of preparation can differ. Most oleogels of low molecular weight (waxes, monoglycerides, and sterols) and ethyl cellulose are prepared by dispersion in heated oil, while oleogels of high molecular weight require dehydration of the emulsion system or solvent exchange. This technique does not change the chemical composition of the oils, which allows them to keep their nutritional value. The properties of oleogels can be designed according to technological needs. Therefore, oleogelation is a future-proof solution that can reduce the consumption of TFA and saturated fatty acids while enriching the diet with unsaturated fatty acids. Oleogels can be named “fats of the future” as a new and healthy alternative for partially hydrogenated fats in foods.

Characteristics of interesterified oils prepared from different substrates and their potential as margarine base stock

Abstract Enzymatic interesterification (EIE) of beef tallow (BT) blended with soybean oil (SO), peanut oil (PO) and canola oil (CO) in different proportions were analyzed in terms of the fatty acid compositions, thermal properties (melting and crystallization behaviors), rheological properties, hardness, crystal, and microstructure to achieve their sufficient utilization. The content of saturated fatty acids in all interesterified oils (IOs) decreased gradually with the increase of plant oil content in base stock. The higher the proportion of plant oil in the base stock, the lower the enthalpy of IOs. All IOs were in the solid-like state, and the hardness of all IOs met the requirements of traditional plastic oil. All IOs were mainly in α and β′ crystal forms and can be used as basic stock for special oils.

Enzymatic synthesis and purification of diacylglycerols from mutton tallow and hemp seed oil

Abstract: Production of preparations based on natural ingredients has become a priority in the cosmetic and pharmaceutical industries. The novelty of the study was to present a possible method for obtaining pure diacylglycerols (DAG) from natural raw materials like fats (hemp seed oil and mutton tallow). We have determined the optimum water content for enzymatic interesterification to shift the reaction equilibrium towards hydrolysis and obtain the highest DAG formation yield. Then, DAG were separated and purified by column chromatography. Raw fats were characterized by fatty acids composition and their positional distribution using gas chromatography. Triacylglycerols, diacylglycerols, monoacylglycerols and free fatty acid contents were determined using gel permeation chromatography. The main fatty acids found in sn-2 MAG of hemp seed oil was linoleic acid, which was followed by oleic acid. Saturated fatty acids in hemp seed oil’s TAG showed preference for the outer sn-1,3 positions. For mutton tallow, the majority of oleic acid were found at sn-2 position. The optimum water content applied during enzymatic interesterification for DAG formation was estimated to be 2.0 % w/w, using the following conditions: 5 % w/w of catalyst, 200 rpm, 60 °C, 6h. DAG yield was 27 %. Purification of the mutton tallow-based and hemp seed oil-based DAG yielded with 83 % w/w and 82 % w/w, respectively. High purity DAG could be applied as effective emulsifiers in food, cosmetic or pharmaceutical emulsions.

Immobilized thermophilic lipases for the interesterification of soybean oil

Vegetable oils are a major source of healthy dietary lipids. However, the physicochemical properties of vegetable oils are varied, which affects their application in foods. Interesterification is a method often used to modify the physicochemical characteristics of oils and fats. Enzymatic interesterification is a process that uses enzymes to exchange the acyl groups among triacylglycerols (TAG); an immobilized lipase is one such enzyme that can be used for interesterification. This study immobilized lipases from two strains of thermophilic bacteria (Geobacillus stearothermophilus (GS) and Anoxybacillus flavithermus (AF)) onto silica beads. These immobilized lipases were used to interesterify soybean oil (SBO) with palmitic acid (PA) to change the physicochemical properties of the oil. High-performance liquid chromatography (HPLC) was used to determine the TAG composition and free fatty acids (FFA) after interesterification. The melting point was measured using differential scanning calorimetry (DSC). Optimum reaction conditions were observed at 70°C, and 24 hrs for all reaction ratios of SBO to PA (1:0.1, 1:0.25, and 1:0.5 (v/v)). The PA significantly decreased after 24 hrs at all reaction ratios, and the TAG concentration was significantly different after interesterification. FFA concentration changed after SBO interesterification, and the melting points of the samples increased from 4°C to 40°C. The results suggest that both immobilized lipases have suitable properties for oil interesterification and industrial applications.

Preparation of human milk fat substitute and improvement of its oxidative stability

1,3-Dioleoyl-2-palmitoylglycerol (OPO) was synthesized by enzymatic interesterification using palm stearin rich in tripalmitin (PPP) and ethyl oleate. Enzymatic interesterification parameters such as temperature, water content, enzyme load, and substrate molar ratio were optimized. High contents of C52 (primarily OPO and its isomeric compounds) production (46.7%) and sn-2 palmitic acid (PA) content of 75.3% were detected. In addition, OPO-human milk fat substitute (HMFS) was blended with coconut, soybean, algal and microbial oils at a weight ratio of 0.70:0.18:0.11:0.004:0.007 to simulate fatty acids in human milk fat (HMF) according to the mathematical model. The main and important fatty acids in the Final-HMFS were within the ranges of those present in HMF. The Final-HMFS could promote the absorption of fats and minerals and the development of retina tissues in infants. The mixture of L-ascorbyl palmitate (L-AP) and vitamin E (VE) resulted in a synergistic antioxidant effect both in OPO-HMFS and OPO-HMFS emulsions. This finding has great significance in improving the quality and extending shelf-life of HMFS.

Design and simulation of a caprylic acid enzymatically modified phosphatidylcholine micelle using a coarse-grained molecular dynamics simulations approach

Computationally simulated micelle models provide useful structural information on the molecular and biological sciences. One strategy to study the self-aggregation process of surfactant molecules that make up a micelle is through molecular dynamics (MD) simulations. In this study, a theoretical approach with a coarse-grained MD simulation (CG-MD) was employed to evaluate the critical micellar concentration (CMC), the micellization process, building a tridimensional (3D) model system of a micelle using data from the experimentally enzymatically modified phospholipids (PL) by phospholipase A1 (PA1). This required enzymatic interesterification of soybean phosphatidylcholine (PC) with caprylic acid, along with purification and characterization by chromatographic techniques to measure the esterified fatty acids and the corresponding PL composition. The number of molecules used in the CG-MD simulation system was determined from the experimental CMC data which was 0.025%. The molecular composition of the system is: 1 C 18:2, 2 C 8:0/8:0, 3 C 8:0/18:3n-9, 4 C 8:0/18:0, 5 C8:0/18:2n-6, 6 C8:0/18:1n-9, and 7 C 8:0/16:0. According to our theoretical results, the micelle model is structurally stable with an average Rg of 3.64 ± 0.10 Å, and might have an elliptical form with a radius of 24.6 Å. Regarding CMC value there was a relationship between the experimental data of the modified PLs and the theoretical analysis by GC-MD, which suggest that the enzymatic modification of PLs does not affect their self-aggregation properties. Finally, the micellar system obtained in the current research can be used as a simple and useful model to design optimal biocompatible nanoemulsions as possible vehicles for bioactive small molecules.Communicated by Ramaswamy H. Sarma

Structured lipids produced from palm-olein oil by interesterification: A controllable lipase-catalyzed approach in a solvent-free system

Palm olein (POL) was modified by enzymatic interesterification with different degrees of acyl migration in a solvent-free packed bed reactor. The fatty acid and acylglycerol composition, isomer content, thermodynamic behavior, and relationship between crystal polymorphism, solid fat content (SFC), crystal microstructure, and texture before and after modification were studied. We found that the increase in sn-2 saturation interesterification was not only due to the generated tripalmitin (PPP) but also caused by acyl migration, and the SFC profiles were changed accordingly. The emergence of high melting point acylglycerols was an important factor accelerating the crystallization rate, further shortening the crystallization induction time, leading to the formation of large crystal spherulites, thereby reducing the hardness. The transformation from the β' to the β form occurred during post-hardening during storage. The isomer content also affected the physicochemical properties of the modified POL.

Glycerol free biodiesel synthesis by application of methyl formate in enzymatic interesterification of rapeseed oil

ABSTRACT The aim of studies was to investigate the process of enzymatic interesterification of rapeseed oil with methyl formate and to determine the influence of three independent variables on the product yield. After evaluating the effectiveness of three enzymatic preparations, the industrial preparation Lipozyme TL IM was selected for the research. Using the surface response methodology, a model describing the dependence of the yield of rapeseed methyl esters on the molar ratio of methyl formate to oil, the amount of catalyst and the duration of the process was created. The following conditions were determined in which the highest yield of rapeseed oil methyl esters was obtained: molar ratio of methyl formate to oil 40:1, process duration 60 h and 14.57% of catalyst Lipozyme TL IM from the amount of oil. Under these conditions, an 81.79% yield of rapeseed oil methyl esters was obtained. Comparative studies of the physical and chemical properties of the product obtained during the interesterification process were performed and their compliance with the requirements of conventional biodiesel standard EN 14214 was evaluated.

Mechanisms of interesterified fat digestibility in a muffin matrix using a dynamic gastric model.

Industrially generated trans-fats have been linked with cardiovascular disease (CVD) and have thus been replaced by interesterified (IE) fats, in foods. Interesterification rearranges fatty acids on the glycerol backbone of a triacylglycerol molecule. However, the impact of IE fat on health is unknown. We recently reported differences in lipid absorption kinetics between IE and rapeseed oil (RO). Here, we investigated the mechanisms underpinning IE fat digestion kinetics in the same muffins baked using an IE fat, non-IE fat [with the same fatty acid composition] and rapeseed oil (RO) under simulated conditions. IE and non-IE fats were largely solid in the gastric phase and strongly associated within the muffin matrix, whereas RO formed liquid droplets which separated from the matrix. No significant difference in lipolysis rates was detected between IE and non-IE fats. The lipolysis of the RO fat was slower, due to long-chain PUFAs. Interesterification itself did not affect digestibility, but the strong interaction between the hard fats and the muffin matrix resulted in extensive creaming of the matrix in the stomach, leading to delayed gastric emptying compared to the RO sample. The rate and extent of lipolysis were determined by the amount of fat available and the structure of the fat. This demonstrates the importance of the physical behaviour of the fats during digestion and provides a mechanistic understanding of the overall lipid digestion of IE fats, which relates to their physiological response.

Enzymatic Interesterification of Palm Stearin, Flaxseed Oil and Cottonseed Stearin to Produce Stable Plastic Fat with Balanced Omega-6 and Omega-3 Fatty Acids.

This study aimed to produce stable plastic fat with desired physicochemical characteristics and ω-6/ω-3 fatty acid ratio (1:1-4:1) from palm stearin (PS), flaxseed oil (FSO) and cottonseed stearin (CS) via enzymatic interesterification (EIE). For the first time, the EIE variables of the blends containing PS, FSO and CS were investigated and optimized through single-factor experiments and response surface design to achieve a high interesterification degree. The optimized enzymatic interesterification conditions were: 60°C, 6 wt% Lipase UM1, and 6 h. Lipase UM1 had a similar effect on ID values with commercial lipases. The EIE improved the compatibility of the lipid blends, with the interesterified product EIE-721 (7:2:1, PS:FSO:CS) being the best candidate base stock for shortening considering its solid fat content, desired ω-6/ω-3 fatty acid ratio, wide melting range, abundant β' form crystal, and compact microstructure. This study provides a strategy to produce balanced ω-6/ω-3 fatty acid plastic fat through enzymatic interesterification and validates the application of Lipase UM1 in the preparation of plastic fat.

Improvement of fatty systems biotechnological interesterification with immobilized enzyme preparation usage

This work research object was fat systems interesterification biotechnology using the Lipozyme TL IM immobilized enzyme preparation. The problem of enzyme preparation activation by moistening with sodium bicarbonate aqueous solution with 7.4 ... 7.7 (3 % wt.) pH was solved in the work. The obtained results made it possible to minimize the interesterification process duration with high-quality product obtaining. The proposed enzyme preparation processing made it possible to reduce the duration of the biointeresterification process in a model fat mixture (palm stearin, coconut and soybean oils in a ratio of 1:1:1, respectively) to 3.5...3.7 hours. The product with high quality indicators, namely up to 0.26 mg KOH/g acid number, up to 0.60 mmol ½ O/kg peroxide number and 1.70 c.u. anisidine number, was obtained as a result. The obtained data can be explained by a fact that effective biocatalysis with lipolytic enzymes as the protein molecules requires the existence of two phases – lipid and water. This fact was provided by the activation parameters justified in the study. The obtained results feature was possibility of enzyme preparation activation, which is not provided under industrial conditions due to the threat of raw materials and finished products hydrolytic processes, which leads to the finished product quality deterioration. The research results made it possible to minimize hydrolytic processes in fat system during interesterification with simultaneous process efficiency increase. From a practical point of view, the discovered activation mechanism made it possible to adjust the enzyme preparation processing conditions in fat systems interesterification technology. The applied aspect of scientific result using was the possibility of improving the typical technological process of fat interesterification

Preparation of functional oils rich in diverse medium and long-chain triacylglycerols based on a broadly applicable solvent-free enzymatic strategy

To address the problems of long reaction times and limited range of adaptation in enzymatic synthesis medium- and long-chain triacylglycerols (MLCTs), a broadly applicable solvent-free enzymatic interesterification strategy was proposed. Candida sp. lipase (CSL) was immobilized on hydrophobic hollow mesoporous silica spheres (HHSS) to construct a biocatalyst designated as CSL@HHSS with a 15.3 % immobilization yield and a loading amount of 94.0 mg/g. The expressed activity and the specific activity were 20.14 U/g and 173.62 U/g, which were 4.6 and 5.6 times higher than that of free CSL, respectively. This biocatalyst demonstrated higher activity, wider applicability, and excellent reusability. Linseed oil, sunflower oil, perilla seed oil, algal oil, and malania oleifera oil were applied as substrates to produce MLCTs with medium-chain triacylglycerols (MCT) catalyzed by CSL@HHSS through interesterification in yields ranging from 69.6 % to 78.0 % within 20 min. Specific fatty acids, including linolenic acid, oleic acid, DHA, and nervonic acid (the first reported), were introduced into MLCT's skeleton, respectively. The structures were finely analyzed and identified by GC and UPLC-MS. The catalytic efficiency value of CSL@HHSS in catalyzing interesterification between linseed oil and MCT (70 ℃, 20 min, lipase 6 wt%) is 0.86 g/g∙min, which is the highest ever reported. This paper presents an effective and sustainable strategy for functional MLCTs production.

Quality Evaluation of Plant Oil Blends Interesterified by Using Immobilized Rhizomucor miehei Lipase

The aim of this research was to evaluate the quality and oxidative stability of enzymatically interesterified plant oil blends. The model plant oil blends consisted of tomato seed oil and coconut oil, which were applied to enzymatic interesterification in the presence of a microbial lipase. To obtain quality characteristics of the enzymatically interesterified oil blends, the following analyses were performed: fatty acids composition and their distribution in internal position (sn-2) in triacylglycerols, oxidative induction time, melting profile, acid value (AV), and peroxide value (PV). The analyzed oil blends contain 6 to 25% monounsaturated fatty acids and 16 to 42% polyunsaturated fatty acids. Additionally, it was noticed that the major monounsaturated fatty acid was oleic acid, with its contribution ranging from 9 to 19%. In most cases, oleic and linoleic acids occupied the sn-2 position of the triacylglycerol molecules, with their contribution reaching 35 to 72% and 34 to 71%, respectively. The enzymatically interesterified oil mixtures were characterized by a relatively long oxidation induction time (41–87 min). Melting profiles of the tested samples revealed the presence of a diversified number of endothermic peaks. The AV and PV of the tested oil blends exceeded 10 mg KOH g−1 fat and 1 meq O2 kg−1 fat, respectively. In conclusion, the tested interesterified plant oil blends are characterized by acceptable thermal and oxidative stability and fatty acid profile.

Physicochemical and textural properties of natural cosmeceutical fats prepared from interesterified oil blends consisting of palm olein and palm kernel oil

Natural oil is commonly used as the base oil of high-end cosmetics. To meet the requirements of general users, the ratio of traditional vegetable oil-based interesterified fats continues to increase in industrial applications. Common interesterified fats are derived from chemical catalysis, which shows high specification requirements for raw materials and catalysts. Although enzymatic interesterification (EIE) is not commonly used in industry, it is far more economical, healthy and environmentally friendly. This study aims to use palm olein (POL) and lauric acid-based palm kernel oil (PKO) as raw materials to produce EIE fats by using Lipozyme TLIM® as the catalyst. The migration of acyl groups during EIE contributed to the modifications of the triacylglycerol compositions, the production of fats with fine crystals due to the accelerated crystallization rate, and improved fat properties such as plasticity, melting profile, viscosity, and spreadability. The interesterified cosmetic base oil showed reduced hydrophobicity and unaltered refractive index with no toxic heavy metal elements introduced during the process.

Enzymatic modification of triglycerides in conventional and surfactant-free microemulsions and in olive oil

Modification of the acylglycerol profile by interesterification of triacyglycerols (TAGs) can improve certain properties of food products. TAGs enriched in palmitic acid (PA) at the sn-2 position and in unsaturated fatty acids at the sn-1,3 position of the glycerol backbone are an important source of nutrients and energy for humans. In the present study, commercially available lipases have been used for the enzymatic interesterification of TAGs to form lipids containing PA and oleic acid (OA). Lipases A and B from Candida antarctica (CaL A and CaL B) and Thermomyces lanuginosus lipase (TLL) were tested in different environments from organic solvents to non-conventional systems such as w/o microemulsions (in the absence and presence of surfactants) and olive oil. The effects of reaction temperature, substrate molar ratio, and external energy were investigated. AOT microemulsions offered an excellent microenvironment for the interesterification reaction, while lecithin microemulsion affected TLL specificity. Surfactant-free microemulsions were also tested in terms of optimization of production process, however lower reaction yields were obtained. For the commercial immobilized CaL B, the ratio PA:triolein affected the reaction yield with the 2:1 ratio showing promising results in organic solvent environment. The main purpose of the present study was to elucidate the appropriate conditions for the enzymatic modification of olive oil. Several factors were tested such as substrates’ ratio, enzyme’s concentration, reaction temperature and external energy input. Overall, the ideal conditions for the formulation of the desired TAGs included the use of 50 mM free PA as substrate, 3 mg of immobilized CaL B/mL of olive oil while the reaction temperature was set at 40 °C and the duration was 72 h.

Enzymatic interesterification of palm olein in a continuous packed bed reactor: Effect of process parameters on the properties of fats and immobilized Thermomyces lanuginosus lipase

Palm olein (POL) was enzymatically interesterified in a continuous packed bed reactor (PBR). The effects of interesterification parameters (flow rates of 4.5–40 g/min and temperatures of 50–90 °C) on the properties of interesterified fats and immobilized Thermomyces lanuginosus lipase (Lipozyme ® TL IM) were investigated. Generally, the percentage of saturated fatty acid at sn-2 remained along with higher flow rates (acyl migration inhibited), while the percentage increased along with higher temperatures and lower flow rates (acyl migration promoted). Solid fat content and crystallization rate of the interesterified fats also showed significant modification with the reaction parameters since these properties are directly influenced by the shift in the triacylglycerol compositions. The occurrence of hydrolysis was also evident during the early stage of interesterification attributed to the lipase's water content, which diminished over time as the water concentration decreased. Under prolonged reaction, the lipase's catalytic activity reduced from 236 U to 65 U, and significant changes in the protein's secondary confirmation were observed. After 100 h of reaction, the α-helix structure decreased (from 23.37% to 16.82%) while β-pleated sheet increased (28.10%–39.59%). The increased fluorescence intensity reflected a significant change in the lipase's tertiary structures.