Lipase-catalyzed incorporation of conjugated linoleic acid into tricaprylin

Three commercially available immobilized lipases, Novozym 435 from Candida antarctica, Lipozyme IM from Rhizomucor miehei, and Lipase PS-C from Pseudomonas cepacia, were used as biocatalysts for the interesterification of conjugated linoleic acid (CLA) ethyl ester and tricaprylin. The reactions were carried out in hexane, and the products were analyzed by gas-liquid chromatography. The effects of molar ratio, enzyme load, incubation time, and temperature on CLA incorporation were investigated. Novozym 435, as compared to Lipozyme IM and Lipase PC-C, showed the highest degree of CLA incorporation into tricaprylin. By hydrolysis with pancreatic lipase, it was found that Lipozyme IM and Lipase PS-C exhibited high selectivity for the sn-1,3 position of the triacylglycerol early in the interesterification, with small extents of incorporation of CLA into the sn-2 position, probably due to acyl migration, at later reaction times. A small extent of sn-1,3 selectivity during interesterification by Novozym 435 was observed.     

Production of margarine fats by enzymatic interesterification with silica-granulated Thermomyces lanuginosa lipase in a large-scale study

Interesterification of a blend of palm stearin and coconut oil (75∶25, w/w), catalyzed by an immobilized Thermomyces lanuginosa lipase by silica granulation, Lipozyme TL IM, was studied for production of margarine fats in a 1- or 300-kg pilot-scale batch-stirred tank reactor. Parameters and reusability were investigated. The comparison was carried out between enzymatic and chemical interesterified products. Experimentally, Lipozyme TL IM had similar activity to Lipozyme IM for the interesterification of the blend. Within the range of 55–80°C, temperature had little influence on the degree of interesterification for 6-h reaction, but it had slight impact on the content of free fatty acids (FFA). Drying of Lipozyme TL IM from water content 6 to 3% did not affect its activity, whereas it greatly reduced FFA and diacylglycerol contents in the products. Lipozyme TL IM was stable in the 1-kg scale reactor at least for 11 batches and the 300-kg pilot-scale reactor at least for nine batches. Due to regiospecificity of the lipase (sn-1,3 specific), enzymatically interesterified products had different fatty acid distribution at sn-2 position from the chemically randomized products, implying the potential nutritional benefits of the new technology. Presented at the 91st American Oil Chemists' Society Annual Meeting in San Diego, April 28, 2000.     

Optimized interesterification of virgin olive oil with a fully hydrogenated fat in a batch reactor: Effect of mass transfer limitations

The lipase‐catalyzed interesterification of virgin olive oil and fully hydrogenated palm oil (FHPO) was studied in a batch reactor operating at 75 °C. The reactions between olive oil {rich in OOO (32.36%), OPO (21.7%) and OLO (11.6%) [L = linoleic; O = oleic; P = palmitic acid]} and the fully hydrogenated fat {(36.5% PSP, 28.8% PPP, 23.2% SPS) [S = stearic acid]} produced semi‐solid fats. For an initial weight ratio of olive oil to FHPO of 60 : 40, the reaction product is a complex mixture of triacylglycerol (TAG) species. The TAG profile of the fat product is time dependent. Because of the high viscosity of the liquid reagent phase, it was important to determine if mass transfer effects were significant. Hence, the reaction was optimized with respect to the type and speed of agitation employed, temperature, use of solvent, and the type of biocatalyst. Three immobilized lipases [from Thermomyces lanuginosus (TL IM), Rhizomucor miehei (RM IM) and Candida antarctica B (Novozym 435)] were compared as catalysts for the interesterification reaction. Equilibrium is reached four times faster (in 1–4 h) with a magnetic stirrer to provide agitation than when agitation is not sufficient, i.e. when orbital agitation is employed. Equilibrium was reached faster with Lipozyme TL IM than with the other two lipases. The effects of all the factors investigated on the composition of the products have also been determined. Semi‐solid fats obtained with the non‐specific Novozym 435 contain levels of unsaturated fatty acid residues on sn‐2 sites that are similar to the products obtained with the 1(3)‐regiospecific enzymes Lipozyme TL IM and RM IM. The chemical properties of the product semi‐solid fat were characterized. The fat prepared using optimal reaction conditions contained 17.20% OPO, 13.61% OOO, 11.09% POP, and 10.35% OSP isomers as the primary products. The induction time obtained in the assay of the oxidative stability of the fat product was 21 h at 98 °C. The lipases Lipozyme TL IM and Novozym 435 were very stable with residual activities of 90 and 100%, respectively, after 15 batch reaction cycles.     

Enzymatic Interesterification of High Oleic Sunflower Oil and Tripalmitin or Tristearin

The objective of this study was to produce low saturated, zero‐trans, interesterified fats with 20 or 30 % saturated fatty acids (SFA) such as C16:0 or C18:0. Tripalmitin (TP) or tristearin (TS) was blended with high oleic sunflower oil (HOSO) at different ratios (0.1:1, 0.3:1, and 0.5:1 [w/w]). Total C16:0 and C18:0 compositions of the resulting TP/HOSO and TS/HOSO blends, respectively, were plotted against blending ratios. Linear interpolation was used to estimate blending ratios that would yield physical blends (PB) with 20 or 30 % SFA. Interesterified blends (IB) were then synthesized from the customized PB using Lipozyme TL IM as the biocatalyst. Total and sn‐2 fatty acid compositions, triacylglycerol (TAG) molecular species, thermal behavior, and oxidative stability of PB and IB were compared. The total fatty acid compositions of PB and IB were similar but fatty acid positional distributions and TAG molecular species composition differed. IB contained 5–10 % more SFA at the sn‐2 position than corresponding PB. Furthermore, interesterification generated mono‐ and disaturated TAG species which resulted in broader melting profiles for IB. However, IB had lower oxidative stability than PB. The reformulation of food products with zero‐trans interesterified fats may be advantageous to the reduction of cardiovascular disease burden in the population.     

Optimization of the synthesis of lower glycerides rich in unsaturated fatty acid residues obtained via enzymatic ethanolysis of sesame oil

The lipases Novozym 435, Lipozyme TL IM and Lipozyme RM IM were employed in the production of lower acylglycerols (LG), i.e. mono‐ (MAG) and diacylglycerols (DAG), rich in unsaturated fatty acids from sesame oil in batch reactors. The effect of the molar ratio of ethanol to fatty acids on the reusability of these immobilized lipases was studied in detail. The effects of pretreatment on lipase activity for ethanolysis were investigated. Glycerol had a strong product inhibition effect on the ethanolysis reaction, and a relatively large excess of ethanol was necessary to remove the glycerol adsorbed on these biocatalysts. The enzymatic activity was drastically reduced by addition of water to the reaction medium. The presence of organic solvents (hexane and acetone) did not favor the production of LG. For the Novozym 435‐catalyzed reaction, optimum conditions were a molar ratio of ethanol to fatty acid residues of 5 : 1, 15 wt‐% lipase and 50 °C. For Lipozyme TL IM, the optimum conditions were a molar ratio of ethanol to fatty acid residues of 5 : 1, 20 wt‐% biocatalyst, and 30 °C. Novozym 435 and Lipozyme TL IM produced LG with molar ratios of unsaturated to saturated fatty acids of 20.4 in 1 h and 25.3 in 5 h, respectively. In the original oil, this ratio was 5. For trials conducted under optimum conditions, the products from the Novozym 435 trials contained 21.8 wt‐% triacylglycerols (TAG), 24 wt‐% DAG and 54.2 wt‐% MAG. The products of the Lipozyme TL IM trials consisted of 12.9 wt‐% DAG and 87.1 wt‐% MAG. No TAG species were detected.     

Interesterification of Lard and Soybean Oil Blends Catalyzed by Immobilized Lipase in a Continuous Packed Bed Reactor

Structured lipids (SL), formulated by blends of lard and soybean oil in different ratios, were subjected to continuous enzymatic interesterification catalyzed by an immobilized lipase from Thermomyces lanuginosus (Lipozyme TL IM) in a continuous packed bed reactor. The original and interesterified blends were examined for fatty acid and triacylglycerol composition, regiospecific distribution, and solid fat content. Blends of lard and soybean oil in the proportions 80:20 and 70:30 (w/w), respectively, demonstrated a fatty acid composition, and proportions of polyunsaturated/saturated fatty acids (PUFA/SFA) and monounsaturated/polyunsaturated fatty acids (MUFA/PUFA), that are appropriate for the formulation of pediatric products. These same blends were suited for this purpose after interesterification because their sn-2 positions were occupied by saturated fatty acids (52.5 and 45.4%, respectively), while unsaturated fatty acids predominantly occupied sn-1,3 positions, akin to human milk fat. Interesterification caused rearrangement of triacylglycerol species.     

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.     

Enzymatic transesterification of triolein and stearic acid and solid fat content of their products

Two systems were investigated and compared as models for making margarine-type fats. Two immobilized lipases, IM60 from Rhizomucor miehei and SP435 from Candida antarctica, were used to catalyze the transesterification of triolein with stearic acid and stearic acid methyl ester, respectively, in n-hexane. The optimal reaction temperature for both enzymes was 55°C at a mole ratio of triolein to acyl donor of 1:2. Equilibria were reached at 18 h for IM60 and 24 h for SP435. Analysis of the overall yield and incorporation of fatty acid at the sn-2 position indicated that the triacylglycerol products contained 38.4 and 16.2% 18:0 for acidolysis and 34.2 and 11.3% for interesterification reactions, respectively, at the 2-position. With SP435, the softest fat was produced after 18 h of incubation, and the hardest after 30 h. For IM60 system, 18 h of incubation gave the most plastic fat.     

Effects of selected substrate forms on the synthesis of structured lipids by two immobilized lipases

Two immobilized lipases, IM 60 from Rhizomucor miehei and SP 435 from Candida antarctica, were used to synthesize structured lipids (SL). Tricaprin and trilinolein were interesterified to produce SL that contained one linoleic acid per triacylglycerol molecule (SL1) and SL with two linoleic acids (SL2). SL1 and SL2 were separated by silver nitrate thin-layer chromatography according to their unsaturation, and the fatty acid at the sn-2 position was determined after pancreatic lipasecatalyzed hydrolysis of SL1 and SL2. With IM 60, 57.7 mol% capric acid and 42.3 mol% linoleic acid were found at the sn-2 position of SL1, while 43.3 mol% capric acid and 56.7 mol% linoleic acid were at the sn-2 position of SL2. The fatty acid at the sn-2 position of SL1 with SP 435 as biocatalyst was 43.6 mol% capric acid and 56.4 mol% linoleic acid, while SL2 contained 56.6 mol% capric acid and 43.4 mol% linoleic acid. Different structural forms of the capric acid-containing substrate (triacylglycerol vs. ethyl ester) and different chainlengths of triacylglycerol were selected to study the substrate selectivity of lipases. Results indicated that SP 435 had some degree of preference for the triacylglycerol form (tricaprin), and IM 60 produced SL more rapidly and reached steady state faster with tricaprin as substrate than with capric acid ethyl ester. For chainlength selectivity, mol% of synthesized SL from tricaprin + trilinolein and tristearin + trilinolein were compared. SP 435 exhibited no apparent preference for either tricaprin or tristearin. However, IM 60 showed a more rapid reaction with tricaprin than with tristearin.     

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.     

Structured lipids: Lipase-catalyzed interesterification of tricaproin and trilinolein

Structured lipids were synthesized by interesterification of trilinolein and tricaproin with sn-1,3-specific (IM 60) and nonspecific (SP 435) lipases. The interesterification reaction was performed by incubating a 1:2 mole ratio of trilinolein and tricaproin in 3 mL hexane at 45°C for the IM 60 lipase from Rhizomucor miehei, and at 55°C for the SP 435 lipase from Candida antarctica. Reaction products were analyzed by reverse-phase high-performance liquid chromatography with an evaporative light-scattering detector. The fatty acids at the sn-2 position were identified after pancreatic lipase hydrolysis and analysis with a gas chromatograph. IM 60 lipase produced 53,5 mol% dicaproyllinolein (total carbon number = C33) and 22.2% monocaproyldilinolein (C45). SP 435 lipase produced 41% C33 and 18% C45. When caproic acid was used in place of tricaproin as the acyl donor, the IM 60 lipase produced 62.9% C33. The effects of variation in mole ratio, temperature, added water, solvent polarity, and time course on the interesterification reaction were also investigated. In the absence of organic solvent, IM 60 lipase produced 52.3% C33.     

Enzymatic Interesterification of Extra Virgin Olive Oil with a Fully Hydrogenated Fat: Characterization of the Reaction and Its Products

The lipase-catalyzed interesterification of extra virgin olive oil (EVOO) and fully hydrogenated palm oil (FHPO) was studied in a batch reactor operating at 75 °C. The compositions of the semi-solid fat products depend on the reaction conditions and the initial ratio of EVOO to FHPO. The dependence of the quasi-equilibrium product TAG profile on the reaction time was determined for initial weight ratios of EVOO to FHPO from 80:20 to 20:80. Lipozyme TL IM, Lipozyme RM IM and Novozym 435 were employed as biocatalysts. The interesterification reaction was optimized with respect to the type and loading of biocatalyst. Equilibrium was approached in the shortest time with Novozym 435 (80% conversion in 4 h). The chemical, physical, and functional properties of the products were characterized. Appropriate choices of the reaction conditions and the initial ratio of EVOO to FHPO lead to TAG with melting profiles and solid fat contents similar to those of commercial products. Differences were observed in the solid fat contents, melting profiles, and oxidative stabilities of the various interesterified products and also between the indicated properties of each category of product and the corresponding physical blend of the precursor reagents.     

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 Emulsions Containing Interesterified Fats Based on Mutton Tallow and Walnut Oil

In this work, modified fats were produced by enzymatic interesterification of mutton tallow with walnut oil. As a result of forcing the fat hydrolysis process by addition of water to the enzymatic preparation (11.5, 13.0, 14.5, 16.0 wt %), additional levels of polar fractions (MAGs, DAGs, and FFAs) were observed. The aim of this work was to evaluate the stability of emulsions of modified fats containing natural emulsifiers resulting from enzymatic interesterification of mutton tallow with walnut oil. The physical‐chemical parameters of obtained fats were determined in this study. Using several methods, the stability of the formed emulsions was also evaluated. The results showed that the fats resulting from interesterification in the presence of Lipozyme RM IM (immobilized lipase from Rhizomucor miehei, Novozymes Bagsvaerd, Denmark) with 13.0, 14.5, and 16.0 wt % of water in the enzymatic preparation could form stable emulsion systems. On the other hand, the emulsion of the interesterification system where the amount of water in the enzymatic preparation was 11.5 % showed very low stability. The number of natural emulsifiers (MAGs and DAGs) that arose after interesterification was insufficient to stabilize the emulsion system. The work has shown the possibility of using interesterified fats as the fat phase. Emulsions formed on the basis of interesterified fats without any additional emulsifiers such as sunflower lecithin had properties comparable to emulsions containing mixed non‐interesterified fat containing additional emulsifier. The natural emulsifiers formed as part of enzymatic interesterification allow formation of stable emulsion systems.     

Preparation of Human Milk Fat Substitutes from Lard by Lipase‐Catalyzed Interesterification Based on Triacylglycerol profiles

Human milk fat substitutes (HMFSs) with triacylglycerol profiles highly similar to those of human milk fat (HMF) were prepared from lard by physical blending followed by enzymatic interesterification. Based on the fatty acid profiles of HMF, different vegetable and single‐cell oils were selected and added to the lard. Blend ratios were calculated based on established physical blending models. The blended oils were then enzymatically interesterified using a 1,3‐regiospecific lipase, Lipozyme RM IM (RML from Rhizomucor miehei immobilized on Duolite ES562; Novozymes A/S, Bagsværd, Denmark), to approximate HMF triacylglycerol (TAG) profiles, particularly with respect to the distribution of palmitic acid in the sn?2 position. The optimized blending ratios were determined to be: lard:sunflower oil:canola oil:palm kernel oil:palm oil:algal oil:microbial oil = 1.00:0.10:0.50:0.13:0.12:0.02:0.02. The optimized reaction conditions were determined to be: enzyme load of 11 wt%, temperature of 60 °C, water content of 3.5 wt%, and reaction time of 3 hours. The resulting product was evaluated for total and sn?2 fatty acids, polyunsaturated fatty acids, and TAG composition. A high degree of similarity was obtained, indicating the great potential of the product as a fat alternative for use in infant formulas.     

Increasing Stearidonic Acid (SDA) in Modified Soybean Oil by Lipase-Mediated Acidolysis

The objective of this work was to synthesize a structured lipid (SL) enriched in stearidonic acid (SDA, C18:4 ω-3), from modified soybean oil (MSO) originally containing ~25% SDA. Low temperature crystallization (LTC) of MSO triacylglycerols (TAG) and free fatty acids (FFA) was performed. The TAG and FFA crystallization products (LTC-TAG and LTC-FFA, respectively) had SDA contents of 48.72 and 60.78%, respectively. Enzymatic acidolysis between MSO and LTC-FFA was studied utilizing Novozym 435 and Lipozyme TL IM as biocatalysts. Substrate molar ratio, incubation time, solvent, and enzyme load were explored. Equilibrium was reached at 96 and 48 h for Novozym 435 and Lipozyme TL IM-catalyzed reactions, respectively. The best conditions from these studies were also applied to the acidolysis of LTC-TAG and LTC-FFA. Utilizing Lipozyme TL IM and solvent free conditions, SLs with SDA contents of 37.61 ± 1.00% (20.86 ± 6.48% at sn-2 position) and 53.46 ± 1.85% SDA (36.37 ± 3.14% at sn-2 position) were obtained from the acidolysis reaction between MSO and LTC-FFA, and LTC-TAG and LTC-FFA, respectively. Compared to the original SDA content of MSO, this process leads to a 52 and 116% increase in SDA content, respectively.     

Lipozyme IM‐catalyzed interesterification for the production of margarine fats in a 1 kg scale stirred tank reactor

Lipozyme IM‐catalyzed interesterification of the oil blend between palm stearin and coconut oil (75/25 w/w) was studied for the production of margarine fats in a 1 kg scale batch stirred tank reactor. Parameters such as lipase load, water content, temperature, and reaction time were investigated. The reusability of Lipozyme IM was also studied under optimized conditions. The interesterification products were monitored by analysis of triacylglycerol profiles, the contents of diacylglycerols, free fatty acids (FFA), and solid fat contents. The contents of some triacylglycerol species, which were categorized by equivalent carbon number (ECN), namely ECN34, 36, 48, and 50, decreased by 6.0, 5.9, 5.8, and 13.7%, respectively, after enzymatic interesterification, similar to the reduction of those species after chemical interesterification, 6.6, 6.0, 7.1, and 12.9%, respectively. On the other hand, those of ECN38, 40, 42, 44, and 46 increased by 1.1, 1.6, 6.8, 16.7, and 6.5%, respectively, in comparison with the increase of those species after chemical interesterification, 0.2, 1.5, 6.5, 17.0, and 9.2%, respectively. Lipase load and reaction time had great influence on the degree of interesterification. A Lipozyme IM load of 6% was required for a reaction of 6 h and at 60 °C, to reach a stable degree of interesterification. Temperature variation in the range of 50—75 °C did not affect the reaction degree as well as the contents of diacylglycerols, but the content of FFA slightly increased with higher temperature. Addition of water to the enzyme increased the contents of diacylglycerols and FFA in the products linearly. However, it had no effect on the degree of interesterification for the first batch when the enzyme was reused. Lipozyme IM was stable in the 10‐batch test after adjusting the water content in the system. The relationship between the content of water in the system and that of FFAs in the products was evaluated and discussed.     

Enzymatic Interesterification of Palm Stearin and Coconut Oil by a Dual Lipase System

Enzymatic interesterification of palm stearin with coconut oil was conducted by applying a dual lipase system in comparison with individual lipase-catalyzed reactions. The results indicated that a synergistic effect occurred for many lipase combinations, but largely depending on the lipase species mixed and their ratios. The combination of Lipozyme TL IM and RM IM was found to generate a positive synergistic action at all test mixing ratios. Only equivalent amount mixtures of Lipozyme TL IM with Novozym 435 or Lipozyme RM IM with Novozym 435 produced a significant synergistic effect as well as the enhanced degree of interesterification. The interesterification catalyzed by Lipozyme TL IM mixed with thermally inactivated immobilized lipase preparations indicated that the carrier property may play an important role in affecting the interaction of two mixed lipases and the subsequent reactions. A dual enzyme system, consisting of immobilized lipases and a non-immobilized one (Lipase AK), in most cases apparently endows the free lipase with a considerably enhanced activity. 70% Lipase AK mixed with 30% immobilized lipase (Lipozyme TL IM, RM IM and Novozym 435) can achieve an increase in activity greater than 100% over the theoretical value when the reaction proceeds for 2 h. The co-immobilization action of the carrier of the immobilized lipases towards the free lipase was proposed as being one of the reasons leading to the synergistic effect and this has been experimentally verified by a reaction catalyzed by a Lipase AK-inactivated preparation. No apparently synergistic effect of the combinations of Lipozyme TL IM and RM IM was observed when the dual enzyme systems applied to the continuous reaction performed in a packed bed reactor. In brief, this work demonstrated the possibility of increasing the reaction rate or enhancing the degree of conversion by employing a dual lipase system as a biocatalyst.     

Operational stability of Thermomyces lanuginosa lipase during interesterification of fat in continuous packed‐bed reactors

The operational stability of a commercial immobilized lipase from Thermomyces lanuginosa (“Lipozyme TL IM”) during the interesterification of two fat blends, in solvent‐free media, in a continuous packed‐bed reactor, was investigated. Blend A was a mixture of palm stearin (POS), palm kernel oil (PK) and sunflower oil (55 : 25 : 20, wt‐%) and blend B was formed by POS, PK and a concentrate of triacylglycerols rich in n‐3 polyunsaturated fatty acids (PUFA) (55 : 35 : 10, wt‐%). The bioreactor operated continuously at 70 °C, for 580 h (blend A) and 390 h (blend B), at a residence time of 15 min. Biocatalyst activity was evaluated in terms of the decrease of the solid fat content at 35 °C of the blends, which is a key parameter in margarine manufacture. The inactivation profile of the biocatalyst could be well described by the first‐order deactivation model: Half‐lives of 135 h and 77 h were estimated when fat blends A and B, respectively, were used. Higher levels of PUFA in blend B, which are rather prone to oxidation, may explain the lower lipase stability when this mixture was used. The free fatty acid content of the interesterified blends decreased to about 1% during the first day of operation, remaining constant thereafter.     

Mono‐Estolide Synthesis from trans‐8‐Hydroxy‐Fatty Acids by Lipases in Solvent‐Free Media and Their Physical Properties

Pseudomonas aeruginosa 42A2 is known to produce two hydroxy‐fatty acids, 10(S)‐hydroxy‐8(E)‐octadecenoic and 7,10(S,S)‐dihydroxy‐8(E)‐octadecenoic acids, when cultivated in a mineral medium using oleic acid as a single carbon source. These compounds were purified, 91 and 96 % respectively, to produce two new families of estolides: trans‐8‐estolides and saturated estolides from the monohydroxylated monomer. trans‐8‐estolides were produced by three different lipases (Novozym 435, Lipozyme RM IM and Lipozyme TL IM) with reaction yields between 68.4 ± 2.1 and 94.7 ± 2.4 % in a solvent‐free medium at 80 °C in 168 h under vacuum. Novozym 435 was found to be the most efficient biocatalyst for both hydroxy‐fatty acids with reaction yields of 71.7 ± 2.3 and 94.7 ± 2.4 %, respectively. Moreover, saturated estolides were also produced from a saturated 10(S)‐hydroxy‐8(E)‐octadecenoic. These estolides were chemically and enzymatically synthesized with Novozym 435, under the previous described reaction conditions with yields of 60.7 ± 2.1 and 71.2 ± 2.3 % respectively. Finally, viscosity, glass transition temperature, decomposition temperatures and enthalpies were determined to characterize both types of estolides. Thermal applications for both types of polyesters were improved since glass transition temperatures were lowered and decomposition temperatures were increased, with respect to their corresponding substrates.