Use of enzymatic transesterified palm stearin-sunflower oil blends in the preparation of table margarine formulation

Palm stearin–sunflower oil (PS:SO) blends, formulated by mixing 40 to 80% palm stearin in increments of 10% (w/w), were subjected to transesterification catalysed by lipases from Pseudomonas sp. and Rhizomucor miehei (Lipozyme 1M 60). The physical properties of the transesterified products were evaluated by slip melting point (SMP), differential scanning calorimetry (DSC), solid fat content (SFC) and X-ray difflaction (XRD) analyses. SMP results indicate that Pseudomonas lipase caused a bigger drop in SMP (33%) in the PS–SO (40:60) blend than the R. miehei-lipase-catalysed reaction blend (13%). The Pseudomonas-catalyzed blends of PS-SO, at 40:60 and 50:50 ratios, showed complete melting at 37 and 40°C, respectively, while the R. miehei-catalyzed PS–SO blend at 40:60 ratio had a residual SFC of 3.9% at 40°C. Pseudomonas lipase also successfully changed the polymorphic form(s) in the unreacted PS–SO mixture from a predominantly β form to a predominantly β′ form in the transesterified blends. However, no changes in polymorphic forms were observed after transesterification with R. miehei lipase (as against to the unreacted PS–SO blends). These results suggest that the Pseudomonas lipase caused a greater randomization and diversification of fatty acids, particularly palmitic acids, in palm stearin with the unsaturated fatty acids from sunflower oil than did R. miehei lipase. Based on the physical characteristics, the Pseudomonas-catalyzed 40:60 and 50:50 PS:SO blends would be the two most suitable blends to be used as table margarine formulations.     

Heat-oxidation stability of palm oil blended with extra virgin olive oil

Rancimat induction time of palm oil (PO), several extra virgin olive oils (EV) and their binary blends have been determined at three different temperatures (120, 130 and 140°C). Analytical composition and oxidation stability of PO/EV blends were found to be a linear combination of the oil partners. Induction time of pure PO was always higher than those of EV oils and blends, in which induction time increased proportionally with the percentage of PO. However, induction time of 80% PO blend was similar to that of pure PO. Fatty acid composition appeared to be the most important factor affecting heat-oxidation stability and a saturated/unsaturated ratio near 1 was the optimally stable composition. Conversely, total phenols had a zero or negative role on the oxidative stability of the blends. Finally, in heat-oxidised oils significant losses of polyunsaturated fatty acids and formation of short-chain fatty acids were recorded.     

High performance liquid chromatographic separation of interesterified palm oil with tributyrin

Short-chain triacylglycerol, tributyrin (glyceryl-sn-1,2,3-tri-butyric acids), was interesterified with palm oil to produce mixtures of structured triacylglycerol (SL-TAG) species as a low-calorie lipid. Lipozyme RM IM from Rhizomucor miehei, known as a 1,3-regioselective immobilized lipase, was used as a catalyst. During 24 h reaction, the reaction mixture was analyzed with Hypersil^®BDSCPS high performance liquid chromatography (HPLC) column to determine neutral lipids composition. Also, the compositional changes of TAG, as well as monoacylglycerol (MAG)/diacylglycerol (DAG) as by-products, were determined. After 24 h, 5.8 g/100 g DAG was found in the reaction mixture, while 89.5 g/100 g TAG was observed. The reaction mixture was also analyzed with Nova-Pak^® C18 and Chrompack Si60 columns for separation of SL-TAG species. It was found that the normal-phase (NP)-HPLC with Chrompack Si60 column readily separated SL-TAG species composed of short- and long-chain acyl residues. Further, mass spectrometer was employed for identifying the separated SL-TAG molecules. SL-TAG containing two butyryl and one long-chain acyl residue (e.g., PBB and OBB) were most abundant in the interesterified product.     

Physical properties of palm kernel olein-anhydrous milk fat mixtures transesterified using mycelium-bound lipase from Rhizomucor miehei

The transesterification activity of mycelium-bound lipase from Rhizomucor miehei on palm kernel olein:anhydrous milk fat (PKO:AMF) blends was investigated. Commercial immobilised R. miehei lipase preparation, Lipozyme IM60 (Novo Nordisk), was used as a comparison. Mixtures of PKO:AMF, at ratios of 100:0, 70:30, 60:40, 50:50 and 0:100, were transesterified using either enzyme in a solvent-free system. The triglyceride (TG) profile, slip melting point, solid fat content, melting thermogram and the polymorphic form of the unreacted and transesterified mixtures were evaluated. Results indicated that transesterification by either enzyme was able to produce an oil mixture with new TG profiles, generally lower slip melting points and solid fat contents. The melting thermograms from differential scanning calorimetry analysis indicated changes in the triglyceride's crystalline composition and an overall shift to lower melting TG. Although the catalytic activities were similar for both lipases, Lipozyme-catalysed mixtures produced higher degrees of transesterification (43–51%) than mycelium-bound lipase-catalysed (22–34%) mixtures. This study also demonstrated that the transesterified PKO:AMF mixture at 70:30 ratio completely melted at 25C, and this meets the melting criteria for fat used in ice cream formulation.     

Crystallization kinetics of palm oil in blends with palm-based diacylglycerol

Crystallization kinetics of palm oil (PO) in the presence of different concentrations (2, 5, 10, 30 and, 50% w/w) of palm-based diacylglycerol (PB-DAG) were investigated over different ranges of crystallization temperatures. Addition of 30 and 50% (w/w) of PB-DAG (high concentrations) increased significantly (P < 0.05) the melting point and crystallization onset while addition of 2 and 5% PB-DAG did not have significant (P > 0.05) effect. PO and PO blends with 2 and 5% of PB-DAG showed crystal transformation at crystallization temperatures (T_Cr) of 26, 26, 26.5 °C, respectively as reflected in corresponding changes of the Avrami parameters at below and above these T_Cr. This was especially evident for the blends containing 2 and 5% of PB-DAG. Individual comparison of induction time (T_i), Avrami exponent (n), Avrami constant (k) and half-time of crystallization (t_1/2) of blends classified under various supercooling ranges based on the supercooling closeness (± 0.1 °C), showed that addition of 5% of PB-DAG in most of the supercooling ranges significantly (P < 0.05) reduced nucleation rate as well as crystal growth velocity of PO. This was reflected in the significantly (P < 0.05) higher T_i and t_1/2 and lower k. Although the presence of 2% of PB-DAG was found to have inhibitory effect on PO crystallization, this effect was not significant (P > 0.05). Mode of crystal growth attributed to n was changed significantly only in presence of 5% of PB-DAG. Furthermore, presence of 10% PB-DAG showed ??'-stabilizing effect on PO. On the other hand, high concentrations of PB-DAG were found to significantly (P < 0.05) reduce T_i as well as t_1/2 and also increase k suggesting their promoting effects on nucleation and crystallization rate of PO even with the close supercoolings. In addition, they changed crystal growth mode of PO. Amongst the different concentrations of PB-DAG investigated, blend containing 50% of PB-DAG as compared to PO, not only, have healthier benefits but also, may have greater potential applications in plastic fat products due to its unique physical properties.     

Production of olive oil enriched with medium chain fatty acids catalysed by commercial immobilised lipases

Structured triacylglycerols, containing medium chain fatty acids, were produced by acidolysis of virgin olive oil with caprylic or capric acid, at a molar ratio of olive oil:fatty acid of 1:2, at 45 °C for 24 h, in solvent-free media or in n-hexane, catalysed by Thermomyces lanuginosa (Lipozyme TL IM), Rhizomucor miehei (Lipozyme RM IM) and Candida antarctica (Novozym 435) immobilised lipases. Incorporations were always greater for capric than for caprylic acid. For both acids, higher incorporations were always attained in solvent-free media: the highest caprylic acid incorporations were obtained with Novozym 435 (25.5 mol%) and Lipozyme RM IM (25.7 mol%), while similar capric acid incorporations were obtained with all biocatalysts (27.1–30.4 mol%).     

Zero trans shortening using rice bran oil, palm oil and palm stearin through interesterification at pilot scale

Fat blends, formulated by mixing refined, bleached and deodorised (RBD) palm oil (PO) or RBD palm stearin (PS) with RBD rice bran oil (RBO) in various ratios were subjected to chemical interesterification (CIE) at pilot scale using sodium methoxide (NaOMe) as catalyst. The resultant interesterified fat was processed through a margarine crystalliser under optimised conditions. The blends before and after CIE were investigated for triacylglycerol (TAG) composition, solid fat content (SFC) and melting characteristics, polymorphic form, fatty acid composition (FAC), bioactive (tocols, sterols, oryzanol) constituents and trans fatty acids (TFA). CIE was found to be very effective in terms of rearrangement of fatty acids (FAs) among TAGs and consequent changes in the physical characteristics. The SFC of the interesterified PS/RBO blends decreased significantly ( P  ≤ 0.05) when compared with those of PO/RBO blends. The interesterified binary blends with 50–60% PS and 40–50% RBO, and 70–80% PO and 20–30% RBO had SFC curves in the range of all-purpose type shortenings. CIE facilitated the formation of β' polymorphic forms. FAC of shortenings prepared using the optimised blends contained 15–20% C_18:2 polyunsaturated fatty acid (PUFA) and no TFA. Total tocol, sterol and oryzanol content of zero trans shortenings were 650–1145, 408–17 583 and 1309–14 430 ppm. CIE using NaOMe did not affect the bioactive constituents significantly ( P  ≤ 0.05).     

Crude palm oil from interspecific hybrid Elaeis oleifera × Elaeis guineensis: Fatty acid regiodistribution and molecular species of glycerides

The composition and structure of triacylglycerols (TAGs) and partial glycerides of crude palm oil obtained from interspecific hybrid Elaeis oleifera × Elaeis guineensis, grown in Colombia, were fully characterised and compared to data obtained by analysing crude African palm oil. Hybridisation appears to substantially modify the biosynthesis of fatty acids (FAs) rather than their assembly in TAGs. In fact, total FAs analysis showed significant differences between these two types of oil, with hybrid palm oil having a higher percentage of oleic acid (54.6 ± 1.0 vs 41.4 ± 0.3), together with a lower saturated fatty acid content (33.5 ± 0.5 vs 47.3 ± 0.1), while the percentage of essential fatty acid, linoleic acid, does not undergo significant changes. Furthermore, 34 TAG types were identified, with no qualitative differences between African and E. guineensis × E. oleifera hybrid palm oil samples. Short and medium chain FAs (8:0, 10:0, 12:0, 14:0) were utilised, together, to build a restricted number of TAG molecular species. Oil samples from the E. guineensis × E. oleifera hybrid showed higher contents of monosaturated TAGs (47.5–51.0% vs 36.7–37.1%) and triunsaturated TAGs (15.5–15.6% vs 5.2–5.4%). The sn-2 position of TAGs in hybrid palm oil was shown to be predominantly esterified with oleic acid (64.7–66.0 mol% vs 55.1–58.2 mol% in African palm oil) with only 10–15% of total palmitic acid and 6–20% of stearic acid acylated in the secondary position. The total amount of diacylglycerols (DAGs) was in agreement with the values of free acidity; DAG types found were in agreement with the representativeness of different TAG species.     

Physicochemical characteristics of soybean oil, palm kernel olein, and their binary blends

Soybean oil (SBO), palm kernel olein (PKO) and their binary blends (containing 5–40% PKO) were studied for their physicochemical characteristics. Decreases in band absorbencies of the resultant Fourier transform infrared spectra were observed in regions attributable to vibrations of the functional groups of unsaturated fatty acids, mainly the =C–H cis stretching at 3009 cm⁻¹and –C=C cis stretching at 1657 cm⁻¹. The solid fat content was measurable in the blends containing 15–40% PKO at 5 and 10 °C, ranging within 4–20% and 2–13%, respectively. The differential scanning calorimetry melting curve for SBO exhibited more complex transition peaks, suggesting a     polymorphic transformation when compared with PKO with a simpler     . Blending of SBO with PKO reduced the complexity caused by the polymorphic transformation, featuring the endotherms that only related to the β' fat crystals.     

Trans-free plastic shortenings from coconut stearin and palm stearin blends

Coconut oil was fractionated to get 40% and 60% stearins (CSt1 and CSt2), blended with a hard fraction (PSt) from palm oil (10–90%) and its suitability as a plastic fat was studied. Slip melting point increased with increase in the level of PSt and ranged from 25.9 to 49.6 °C. CSt2 did not contain any solids above 20 °C but increased on incorporating PSt and showed the wider melting range required for plastic fats. Melting profiles of the blends containing 30% and 40% PSt were comparable with that of a commercial bakery shortening. Triglyceride composition showed that the blends contained lower and higher molecular weight triglycerides desirable for plastic fats. FTIR spectra showed a distinct peak at 966 cm⁻¹ characteristic of trans fatty acids in commercial shortening while the peak was absent in blends. This study showed that the blends containing 60–70% CSt with PSt are suitable as trans-free plastic fats.     

Biocatalyzed acidolysis of olive oil triacylglycerols with 9c,11t and 10t,12c isomers of conjugated linoleic acid

Lipase catalyzed acidolysis of triacylglycerols (TAG) of olive oil with 9c,11t and 10t,12c isomers of conjugated linoleic acid (CLA) in an organic solvent was studied. The CLA isomers were first obtained in good yield starting from sunflower oil. The acidolysis reactions were carried out with two immobilized lipase, an sn-1,3-regiospecific one from Rhizomucor miehei and a nonregiospecific one from Candida antartica, in order to valuate not only the total incorporation of CLA isomers in olive TAG but also the distribution of the cited isomers in the three sn positions of TAG molecules. The effect of reaction time was also investigated; in fact two series of reactions, with the two lipases, were carried out for 24, 48 and 72 h. The TAG fractions relative to the starting olive oil TAG (OOTAG) and to the samples obtained from the acidolysis reactions were analyzed to obtain the total and positional fatty acid percentage compositions. The results show that after 24 h of reaction, high levels of CLA isomers were incorporated in OOTAG using Lipozyme IM and that slightly higher values were obtained by increasing the reaction time; Novozym 435 was less effective in catalyzing the incorporation of CLA isomers and CLA isomers were incorporated in OOTAG to a lesser degree. The results of stereospecific analysis of TAG fractions showed that, at every reaction time, the CLA isomers were incorporated mainly in sn-1 and sn-3 positions, as expected on the basis of the enzyme regiospecificity. As regards the TAG sn-2 position, the incorporation of CLA isomers was not observed after 24 h, but after 48 and 72 h; this occurrence was probably due to isomerization phenomena or regiospecificity loss after extended reaction times.     

Incorporation of selected long-chain fatty acids into trilinolein and trilinolenin

The effect of chain length, number of double bonds, the location and geometry of double bonds, the reaction conditions, and reactivity of different lipases on the incorporation of selected long-chain fatty acids (LCFA) into triacylglcerols, such as trilinolein (tri C18:2) and trilinolenin (tri C18:3) is examined. This study also discusses reasons behind different degrees of incorporation of selected LCFA into tri C18:2 or tri C18:3 on a molecular basis. Five lipases, namely Candida antarctica (Novozyme-435), Mucor miehei (Lipozyme-1M), Pseudomonas sp. (PS-30), Aspergillus niger (AP-12), and Candida rugosa (AY-30) were screened for their effect on catalyzing the acidolysis of trilinolein (tri C18:2) or trilinolenin (tri C18:3) with selected C18, C20 and C22 fatty acids (FA). Incorporation of a mixture of C18 FA into trilinolein, using Pseudomonas sp., the most effective lipase, was in the order of SA > OA > GLA > ALA > CLA. Meanwhile, the degree of n-6 FA incorporation into tri C18:2 with Pseudomonas sp. was in the order of GLA > AA > CLA. The order of incorporation of n-3 FA into trilinolein using lipases from C. antarctica and M. miehei was ALA > EPA > DPA > DHA.     

Utilization of interesterified oil blends in the production of frankfurters

Ten treatments of frankfurters were produced with interesterified oil and oil blends (palm oil, palm stearin, cottonseed oil, hazelnut oil and their mixtures) and were compared to control, produced with all animal fat. Addition of interesterified oil and oil blends affected (p < 0.05) the moisture and fat content and pH values of frankfurters. According to the colour measurements, the brightness value (L^∗) of most of the samples with interesterified oil and oil blends were higher (p < 0.05) than the control. The fatty acid composition of frankfurters was modified. The PUFA/SFA values of frankfurters were increased due to the presence of interesterified oil and oil blends in the formulation. Frankfurters with 100% interesterified cottonseed oil or with interesterified oil blends with 66.6% and 83.4% cottonseed oil had PUFA/SFA ratio higher than 0.4 and are considered better than all others from the health point of view. Frankfurters produced with 100% interesterified cottonseed and hazelnut oil or with interesterified hazelnut oil blends had the same (p > 0.05) scores for sensory attributes with the control, while all other treatments were also acceptable.     

Thermal behavior of concentrated oil-in-water emulsions based on soybean oil and palm kernel olein blends

Droplet size distribution and thermal behavior of concentrated oil-in-water emulsions based on soybean oil (SBO)/palm kernel olein (PKO) blends were investigated. The emulsions were prepared using 70% (wt./wt.) oil blends of SBO/PKO as dispersed phases and stabilized by egg yolk. An increase in PKO level (0–40% wt./wt.) in the oil dispersed phase volume fraction caused significant increases (p < 0.05) in volume-weighted mean diameter (d_4,3). The DSC data suggested that crystallization of the emulsions was induced by a ‘template effect’ of yolk constituents via a surface heterogeneous nucleation. Emulsions with 0–20% (wt./wt.) PKO levels in the dispersed phase demonstrated a good cool–heat stability even after three successive thermal cycles (from 50 °C to ?70 °C at 10 min/°C). After the first thermal cycle, emulsions with 30% and 40% PKO levels in the oil dispersed phase were destabilized due to strong coalescence and crystallized via volume-surface heterogeneous nucleation. The unstable emulsions were attributable to high level of saturated triacylglycerols from PKO, with high droplet size characteristic, causing them to be more prone to partial coalescence.     

Improvement of palm oil and sunflower oil blends by enzymatic interestrification

Palm oil (PO) and sunflower oil (SFO) blends with varying proportions were subjected to enzymatic interesterification (EIE) using a 1,3‐specific immobilised lipase. The interesterified blends were evaluated for their slip melting point (SMP), solid fat content (SFC) at 10–40 °C, p‐anisidine value, peroxide value, free fatty acids (FFA), induction period of oxidation at 110 °C (IP110) and composition of fatty acids by gas chromatography. Under EIE treatment, the blends of PO and SFO in different proportions (20:80, 40:60, 50:50, 60:40 and 80:20) had saturated and unsaturated fatty acid content in the range of 37.6–52.0% and 48.0–62.4%, respectively. The blends showed a considerable reduction in their SFC, SMP, peroxide value and oxidative stability at 110 °C, but presented increase in FFA and p‐anisidine value. The optimum condition for minimising the fatty acid in oil was obtained, at 64 °C, using 8.9% enzyme and 3 h reaction time.     

Glycidyl esters in refined palm (Elaeis guineensis) oil and related fractions. Part I: Formation mechanism

Glycidyl esters (GE) are process contaminants generated during the deodorisation step of edible oil refining. In particular, GE are found in high abundance in refined palm oil. Palm oil is unique in that it contains a high amount of diacylglycerols (DAG, 4-12%). In the present study, a series of model reactions mimicking palm oil deodorisation has been conducted with pure tri-, di- and mono-acylglycerols (MAG). Results showed that GE are formed from DAG and MAG, but not from TAG, at temperatures (T) above 200 °C. Our observations suggest that GE are formed predominantly by intramolecular elimination of a fatty acid from DAG. In addition, isomers of GE, formed from DAG heated at T > 140 °C, were identified as oxopropyl esters. These new isomers were found to represent approximately 10% of GE levels in refined palm oil. Based on these considerations, the final GE content of palm oil could be limited by reducing DAG levels before oil processing and minimising deodorisation temperatures.     

Enzymatic interesterification of palm and coconut stearin blends

Hard fractions of palm oil and coconut oil, blended in the ratios of 90:10, 85:15, 80:20 and 75:25, were interesterified for 8 h using Lipozyme TL IM. Major fatty acids in the blends were palmitic acid (41.7–48.4%) and oleic acid (26.2–30.8%). Medium‐chain fatty acids accounted for 4.5–13.1% of the blends. After interesterification (IE), slip melting point was found to decrease from 44.8–46.8 °C to 28.5–34.0 °C owing to reduction in solids content at all temperatures. At 37.5 °C, the blends containing 25% coconut stearins had 17.4–19% solids, which reduced to 0.4–1.5% on IE, and the slip melting point (28.6 and 28.8 °C) indicated their suitability as margarine base. The reduction in solid fat index of the interesterified fats is attributed to the decrease in high‐melting triacylglycerols in palm oil (GS₃ and GS₂U type) and increase in triolein (GU₃) content from 1 to 9.2%. Retention of tocopherols and β‐carotene during IE was 76 and 60.1%, respectively, in 75:25 palm stearin and coconut stearin blend.     

Influence of dietary oils and protein level on pork quality. 2. Effects on properties of fat and processing characteristics of bacon and frankfurter-style sausages

Palm kernel oil (PKO) and palm oil (PO) are used in tropical countries as cheaper substitutes for conventional feed sources such as soya bean oil (SBO) but little is known about their effects on meat quality. This study, therefore, evaluated the effects of these three dietary oils on the fatty acid composition (FA) of pork fat and the qualities of belly bacon and frankfurter sausage. The 3×2 factorial design also included high and low dietary protein. Total cooking loss, water loss and fat losses were determined in frankfurter sausages at chopping temperatures from 2 to 24°C. PKO resulted in a poor P:S ratio (0.34) and a relatively hard fat (slip point 32.8°C), but resulted in bacon with a higher tensile cohesive force and more high quality slices, judged subjectively. PO had a fatty acid composition closer to the SBO control, a better P:S ratio than PKO (0.48) and softer fat. There was a trend for total cooking losses and fat losses to be higher in PKO compared with PO and SBO at all chopping temperatures, suggesting that the firmest, most saturated fat (PKO) was least suitable for frankfurter production. The low protein diet increased the concentration of saturated fatty acids and increased fat firmness but its effect on fatty acid composition and other properties were less marked than those of oil type.     

Influence of dietary oils and protein level on pork quality. 1. Effects on muscle fatty acid composition, carcass, meat and eating quality

This study evaluated the effects of three dietary oils - palm kernel (PKO), palm (PO) and soyabean (SBO) - and two protein levels - high (HP) and low (LP) in a 3×2 factorial design involving 60 pigs on growth performance, muscle fatty acid composition and content, carcass, meat and eating qualities. Oil type did not have a significant effect on growth and carcass quality. PKO significantly reduced the polyunsaturated (PUFA) to saturated (SFA) fatty acid (P:S) ratio in longissimus muscle (P<0.001). PKO increased the concentrations of lauric (12:0), myristic (14:0), palmitic (16:0) and stearic (18:0) fatty acids and decreased linoleic acid (18:2). The LP diet increased intramuscular fat (IMF) from 1.7g/100g muscle in HP to 2.9g/100g (P<0.001), increased tenderness by 0.6 units (P<0.01) and juiciness by 0.5 units (P<0.01) on the 1-8 scale, but at the expense of lower daily weight gain (P<0.01), lower feed conversion efficiency (P<0.01), reduced P:S ratio (P<0.001) and increased lipid oxidation (P<0.01). The results suggest that PKO and PO could be used in tropical developing countries as cheaper alternatives to SBO for the production of good quality and healthy pork, but their limits of inclusion need to be determined.     

Extraction of β-carotenes from palm oil mesocarp using sub-critical R134a

Sub-critical extraction of palm oil from palm mesocarp using R134a solvent was conducted via the dynamic mode to investigate the ability of R134a to extract β-carotene. The yield of palm oil and the solubility of β-carotene were investigated at 40, 60 and 80 °C and pressure range from 45–100 bar. The extracted oil was analysed for β-carotene content using UV–Vis spectrophotometry. The results showed that palm oil yield increased with pressure and temperature. The maximum solubility of β-carotene was obtained at 100 bar and 60 °C while the lowest solubility occurred at 80 bar and 40 °C. The higher concentration of extracted β-carotene ranging from 330–780 ppm as compared to that achieved through conventional palm oil processing indicates that extraction of β-carotene using R134a is viable.