Medium- and long-chain triacylglycerol (MLCT) is a modified lipid containing medium- chain (C6-C12) and long-chain fatty acids (C14-C24) in the same triacylglycerol (TAG) molecule. It can be produced either through enzymatic (with 1,3 specific or nonspecific enzyme) or chemical methods. The specialty of this structured lipid is that it is metabolized differently compared to conventional fats and oils, which can lead to a reduction of fat accumulation in the body. Therefore, it can be used for obesity management. It also contains nutritional properties that can be used to treat metabolic problems. This review will discuss on the health benefits of MLCT, its production methods especially via enzymatic processes and its applications in food industries.
In this study, the methods for extraction and purification of miraculin from Synsepalum dulcificum were investigated. For extraction, the effect of different extraction buffers (phosphate buffer saline, Tris-HCl and NaCl) on the extraction efficiency of total protein was evaluated. Immobilized metal ion affinity chromatography (IMAC) with nickel-NTA was used for the purification of the extracted protein, where the influence of binding buffer pH, crude extract pH and imidazole concentration in elution buffer upon the purification performance was explored. The total amount of protein extracted from miracle fruit was found to be 4 times higher using 0.5M NaCl as compared to Tris-HCl and phosphate buffer saline. On the other hand, the use of Tris-HCl as binding buffer gave higher purification performance than sodium phosphate and citrate-phosphate buffers in IMAC system. The optimum purification condition of miraculin using IMAC was achieved with crude extract at pH 7, Tris-HCl binding buffer at pH 7 and the use of 300 mM imidazole as elution buffer, which gave the overall yield of 80.3% and purity of 97.5%. IMAC with nickel-NTA was successfully used as a single step process for the purification of miraculin from crude extract of S. dulcificum.
The lipase production ability of a newly isolated Acinetobacter sp. in submerged (SmF) and solid-state (SSF) fermentations was evaluated. The results demonstrated this strain as one of the rare bacterium, which is able to grow and produce lipase in SSF even more than SmF. Coconut oil cake as a cheap agroindustrial residue was employed as the solid substrate. The lipase production was optimized in both media using artificial neural network. Multilayer normal and full feed forward backpropagation networks were selected to build predictive models to optimize the culture parameters for lipase production in SmF and SSF systems, respectively. The produced models for both systems showed high predictive accuracy where the obtained conditions were close together. The produced enzyme was characterized as a thermotolerant lipase, although the organism was mesophile. The optimum temperature for the enzyme activity was 45°C where 63% of its activity remained at 70°C after 2 h. This lipase remained active after 24 h in a broad range of pH (6-11). The lipase demonstrated strong solvent and detergent tolerance potentials. Therefore, this inexpensive lipase production for such a potent and industrially valuable lipase is promising and of considerable commercial interest for biotechnological applications.
The oxidative and thermal stability of low diglycerides palm oil produced via silica treatment (sPO) and enzymatic treatment (ePO) compared with standard quality palm oil (SQ) and premium quality palm oil (PQ) was investigated. Both of the oils displayed better oxidative stability compared with SQ as well as significantly higher (P < 0.05) thermal resistance and oxidative strength than SQ and PQ due to lower amounts of partial glycerides. Although the initial induction periods (IPs) of sPO and ePO were significantly lower compared with SQ and PQ, both the oils showed slower drops in their IP values. The darkening effect after frying was significantly (P < 0.05) slower in sPO compared with SQ, PQ, and ePO. Besides, there is no difference p > 0.05 in the rate of FFA formation between sPO and PQ. The anisidine value and peroxide values were lowest in sPO, followed by ePO, PQ, and SQ.
Coconut oil is a rich source of beneficial medium chain fatty acids (MCFAs) particularly lauric acid. In this study, the oil was modified into a value-added product using direct modification of substrate through fermentation (DIMOSFER) method. A coconut-based and coconut-oil-added solid-state cultivation using a Malaysian lipolytic Geotrichum candidum was used to convert the coconut oil into MCFAs-rich oil. Chemical characteristics of the modified coconut oils (MCOs) considering total medium chain glyceride esters were compared to those of the normal coconut oil using ELSD-RP-HPLC. Optimum amount of coconut oil hydrolysis was achieved at 29% moisture content and 10.14% oil content after 9 days of incubation, where the quantitative amounts of the modified coconut oil and MCFA were 0.330 mL/g of solid media (76.5% bioconversion) and 0.175 mL/g of solid media (53% of the MCO), respectively. MCOs demonstrated improved antibacterial activity mostly due to the presence of free lauric acid. The highest MCFAs-rich coconut oil revealed as much as 90% and 80% antibacterial activities against Staphylococcus aureus and Escherichia coli, respectively. The results of the study showed that DIMOSFER by a local lipolytic G. candidum can be used to produce MCFAs as natural, effective, and safe antimicrobial agent. The produced MCOs and MCFAs could be further applied in food and pharmaceutical industries.
Separation of 1,2(2,3)- and 1,3-positional isomers of diacylglycerols (DAG) from vegetable oils by reversed-phase high-performance liquid chromatography (RP-HPLC) is investigated. The method is based on isocratic elution using 100% acetonitrile and UV detection at 205 nm. The following elution order of DAG molecular species is identified: 1,3-dilinolein < 1,2-dilinolein < 1,3-dimyristin < 1-oleoyl-3-linoleoyl-glycerol < 1,2-dimyristoyl-rac-glycerol < 1(2)-oleoyl-2(3)-linoleoyl-glycerol < 1-linolenoyl-3-stearoyl-glycerol < 1(2)-linolenoyl-2(3)-stearoyl-glycerol < 1,3-diolein < 1-palmitoyl-3-oleoyl-glycerol < 1,2-dioleoyl-sn-glycerol < 1(2)-palmitoyl-2(3)-oleoyl-glycerol < 1-linoleoyl-3-stearoyl-glycerol < 1,3-dipalmitin < 1(2)-linoleoyl-2(3)-stearoyl-glycerol < 1-oleoyl-3-stearoyl-glycerol < 1,2-dipalmitoyl-rac-glycerol < 1-palmitoyl-3-stearoyl-sn-glycerol < 1,3-distearin < 1,2-distearoyl-rac-glycerol. Linearity is observed over three orders of magnitude. Limits of detection and quantitation range 0.2-0.7 microg/mL for 1,3-dilinolein to 0.6-1.9 microg/mL for 1,2-dioleoyl-sn-glycerol, respectively. Precision and accuracy of the method are also demonstrated. The method is developed to separate mixtures of DAG molecular species produced from edible oils.
Mycelium-bound lipase (MBL) was prepared using a strain of Geotrichum candidum isolated from local soil. At the time of maximum lipase activity (54 h), the mycelia to which the lipase was bound were harvested by filtration and centrifugation. Dry MBL was prepared by lyophilizing the mycelia obtained. The yield of MBL was 3.66 g/l with a protein content of 44.11 mg/g. The lipase activity and specific lipase activity were 22.59 and 510 U/g protein, respectively. The moisture content of the MBL was 3.85%. The activity of free (extracellular) lipase in the culture supernatant (after removal of mycelia) was less than 0.2 U/ml. The MBL showed selectivity for oleic acid over palmitic acid during hydrolysis of palm olein, indicating that the lipase from G. candidum displayed high substrate selectivity for unsaturated fatty acid containing a cis-9 double bond, even in crude form. This unique specificity of MBL could be a direct, simple and inexpensive way in the fats and oil industry for the selective hydrolysis or transesterification of cis-9 fatty acid residues in natural triacylglycerols.
Soy sauce fermentation was simulated in a laboratory and subjected to 10min of sonication. A full factorial design, including different cycles, probe size, and amplitude was used. The composition of 17 free-amino acids (FAAs) was determined by the AccQ-Tag method with fluorescent detection. Main effect plots showed total FAAs extraction was favoured under continuous sonication at 100% amplitude using a 14mm diameter transducer probe, reaching 1214.2±64.3mg/100ml of total FAAs. Moreover, after 7days of fermentation, sonication treatment caused significantly higher levels (p<0.05) of glutamic acids (343.0±22.09mg/100g), total FAAs (1720.0±70.6mg/100g), and essential FAAs (776.3±7.0mg/100g) 3days sooner than the control. Meanwhile, enzymatic and microbial behaviours remained undisturbed. Collectively, the sonication to moromi resulted in maturation 57% faster than the untreated control.
An optimized refining process for kenaf seed oil was conducted. The 3-monochloro-1,2-propanediol (3-MCPD) contents, triacylglycerol composition, fatty acids composition, bioactive compounds, phosphorus contents, and oxidation indexes of the kenaf seed oil during each stage of the refining process were determined. The results showed that there was no detected 3-MCPD ester in kenaf seed oil throughout the refining process. Deodorization had slightly increased the 2-MCPD ester (9.0 μg/kg) and glycidyl ester (54.8 μg/kg). Oleic (36.53%) and linoleic acids (36.52%) were presented in the largest amount in the refined kenaf seed oil, and triacylglycerols contributed to 99.96% in the oil. There was a removal of 31.6% of phytosterol content and 17.1% of tocopherol and tocotrienol contents in kenaf seed oil after refining. The refining process was totally removed the hydroperoxides, 93% of free fatty acids and 98.8% of phosphorus content in kenaf seed oil.
Phospholipid composition in the milk fat globule membrane (MFGM) fluctuates during the entire lactation period in order to suit the growing needs of newborn infants. The present study elucidated and relatively quantified phospholipid molecular species extracted from human milk (HM), mature human milk (MHM), and infant formulas (with or without MFGM supplementation) using hydrophilic liquid chromatography-electrospray ionization-ion trap-time of flight-mass spectrometry (HILIC-ESI-IT-TOF-MS) system. Principal component analysis was used to clarify the differences between phospholipid composition in HM, MHM, and infant formulas. HM and MHM contained high concentrations of sphingomyeline (HM: 107.61 μg/mL, MHM: 227.18 μg/mL), phosphatidylcholine (HM: 59.96 μg/mL, MHM: 50.77 μg/mL), and phosphatidylethanolamine (PE) (HM: 25.24 μg/mL, MHM: 31.76 μg/mL). Significant concentrations (<300 ng/mL) of arachidonic, eicosapentanoic, and docosahexanoic acids were found to esterify to PE in HM and MHM. Meanwhile, all infant formulas were found to contain high concentrations of phosphatidic acids indicating the possibility of degradation of the fortified MFGM either during processing or storage of the infant formulas.
The formation of 3-monochloropropane-1,2-diol (3-MCPD) esters in refined palm oil during deodorisation is attributed to the intrinsic composition of crude palm oil. Utilising D-optimal design, the effects of the degumming and bleaching processes on the reduction in 3-MCPD ester formation in refined palm oil from poor-quality crude palm oil were studied relative to the palm oil minor components that are likely to be their precursors. Water degumming remarkably reduced 3-MCPD ester formation by up to 84%, from 9.79 mg/kg to 1.55 mg/kg. Bleaching with synthetic magnesium silicate caused a further 10% reduction, to 0.487 mg/kg. The reduction in 3-MCPD ester formation could be due to the removal of related precursors prior to the deodorisation step. The phosphorus content of bleached palm oil showed a significant correlation with 3-MCPD ester formation.
Structured lipid such as medium-and long-chain triacylglycerol (MLCT) is claimed to be able to suppress body fat accumulation and be used to manage obesity. Response surface methodology (RSM) with four factors and three levels (+1,0,-1) faced centered composite design (FCCD) was employed for optimization of the enzymatic interesterification conditions of palm-based MLCT (P-MLCT) production. The effect of the four variables namely: substrate ratio palm kernel oil: palm oil, PKO:PO (40:60-100:0 w/w), temperature (50-70 °C), reaction time (0.5-7.5 h) and enzyme load (5-15 % w/w) on the P-MLCT yield (%) and by products (%) produced were investigated. The responses were determined via acylglycerol composition obtained from high performance liquid chromatography. Well-fitted models were successfully established for both responses: P-MLCT yield (R (2) = 0.9979) and by-products (R (2) = 0.9892). The P-MLCT yield was significantly (P 0.05). Substrate ratio PKO: PO (100:0 w/w) gave the highest yield of P-MLCT (61 %). Nonetheless, substrate ratio of PKO: PO (90:10w/w) was chosen to improve the fatty acid composition of the P-MLCT. The optimized conditions for substrate ratio PKO: PO (90:10 w/w) was 7.26 h, 50 °C and 5 % (w/w) Lipozyme TLIM lipase, which managed to give 60 % yields of P-MLCT. Up scaled results in stirred tank batch reactor gave similar yields as lab scale. A 20 % increase in P-MLCT yield was obtained via RSM. The effect of enzymatic interesterification on the physicochemical properties of PKO:PO (90:10 w/w) were also studied. Thermoprofile showed that the P-MLCT oil melted below body temperature of 37 °C.
Diacylglycerol (DAG), which has health-enhancing properties, is sometimes added to bakery shortening to produce baked products with enhanced physical functionality. Nevertheless, the quantity present is often too little to exert any positive healthful effects. This research aimed to produce bakery shortenings containing significant amounts of palm diacyglycerol (PDG). Physicochemical, textural and viscoelastic properties of the PDG bakery shortenings during 3 months storage were evaluated and compared with those of commercial bakery shortening (CS).
The purpose of this study was to optimize the parameters involved in the production of water-soluble phytosterol microemulsions for use in the food industry. In this study, response surface methodology (RSM) was employed to model and optimize four of the processing parameters, namely, the number of cycles of high-pressure homogenization (1-9 cycles), the pressure used for high-pressure homogenization (100-500 bar), the evaporation temperature (30-70 degrees C), and the concentration ratio of microemulsions (1-5). All responses-particle size (PS), polydispersity index (PDI), and percent ethanol residual (%ER)-were well fit by a reduced cubic model obtained by multiple regression after manual elimination. The coefficient of determination (R(2)) and absolute average deviation (AAD) value for PS, PDI, and %ER were 0.9628 and 0.5398%, 0.9953 and 0.7077%, and 0.9989 and 1.0457%, respectively. The optimized processing parameters were 4.88 (approximately 5) homogenization cycles, homogenization pressure of 400 bar, evaporation temperature of 44.5 degrees C, and concentration ratio of microemulsions of 2.34 cycles (approximately 2 cycles) of high-pressure homogenization. The corresponding responses for the optimized preparation condition were a minimal particle size of 328 nm, minimal polydispersity index of 0.159, and <0.1% of ethanol residual. The chi-square test verified the model, whereby the experimental values of PS, PDI, and %ER agreed with the predicted values at a 0.05 level of significance.
Diacylglycerol (DAG) and triacylglycerol (TAG) as responses on optimization of DAG production using a dual response approach of response surface methodology were investigated. This approach takes the molecular equilibrium of DAG into account and allows for the optimization of reaction conditions to achieve maximum DAG and minimum TAG yields. The esterification reaction was optimized with four factors using a central composite rotatable design. The following optimized conditions yielded 48 wt % DAG and 14 wt % TAG: reaction temperature of 66.29 degrees C, enzyme dosage of 4 wt %, fatty acid/glycerol molar ratio of 2.14, and reaction time of 4.14 h. Similar results were achieved when the process was scaled up to a 10 kg production in a pilot packed-bed enzyme reactor. Lipozyme RM IM did not show any significant activity losses or changes in fatty acid selectivity on DAG synthesis during the 10 pilot productions. However, lipozyme RM IM displayed higher selectivity toward the production of oleic acid-enriched DAG. The purity of DAG oil after purification was 92 wt %.
Mycelium-bound lipase (MBL), from a locally isolated Geotrichum candidum strain, was produced and characterized as a natural immobilized lipase. A time course study of its lipolytic activity in 1 L liquid broth revealed the maximum MBL activity at 4 h for mycelium cells harvested after 54 h. The yield and specific activity of MBL were 3.87 g/L dry weight and 508.33 U/g protein, respectively, while less than 0.2 U/mL lipase activity was detected in the culture supernatant. Prolonged incubation caused release of the bound lipase into the growth medium. The growth pattern of G. candidum, and production and properties of MBL were not affected by the scale. The stability of mycelia harboring lipase (MBL), harvested and lyophilized after 54 h, studied at 4 °C depicted a loss of 4.3% and 30% in MBL activity after 1 and 8 months, while the activity of free lipase was totally lost after 14 days of storage. The MBL from G. candidum displayed high substrate selectivity for unsaturated fatty acids containing a cis-9 double bond, even in crude form. This unique specificity of MBL could be a direct, simple and inexpensive way in the fats and oil industry for the selective hydrolysis or transesterification of cis-9 fatty acid residues in natural triacylglycerols.
Structured lipid medium- and long-chain triacylglycerols (MLCT) are claimed to be able to manage obesity. The present study investigated the body fat influence of enzymatically interesterifed palm-based medium- and long-chain triacylglycerols (P-MLCT) on diet-induced obesity (DIO) C57BL/6J mice compared with commercial MLCT oil (C-MLCT) and a control, which was the non enzymatically modified palm kernel and palm oil blend (PKO-PO blend). It also investigated the low fat and high fat effects of P-MLCT. DIO C57BL/6J mice were fed ad libitum with low fat (7%) and high fat (30%) experimental diets for 8 weeks before being sacrificed to obtain blood serum for analysis. From the results, there is a trend that P-MLCT fed mice were found to have the lowest body weight, body weight gain, total fat pad accumulation (perirenal, retroperitoneal, epididymal and mesenteric), total triglyceride levels and efficiency in controlling blood glucose level, compared with C-MLCT and the PKO-PO blend in both low fat and high fat diets. Nevertheless, the PKO-PO blend and P-MLCT caused significantly (P < 0.05) higher total cholesterol levels compared to C-MLCT. P-MLCT present in low fat and high fat dosage were shown to be able to suppress body fat accumulation. This effect is more prominent with the low fat dosage.
The stearin fraction of palm-based diacylglycerol (PDAGS) was produced from dry fractionation of palm-based diacylglycerol (PDAG). Bakery shortening blends were produced by mixing PDAGS with either palm mid fraction, PMF (PDAGS/PMF), palm olein, POL(PDAGS/POL) or sunflower oil, SFO (PDAGS/SFO) at PDAGS molar fraction of XPDAGS=0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%. The physicochemical results obtained indicated that C16:0 and C18:1 were the dominant fatty acids for PDAGS/PMF and PDAGS/POL, while C18:1 and C18:2 were dominant in the PDAGS/SFO mixtures. SMP and SFC of the PDAGS were reduced with the addition of PMF, POL and SFO. Binary mixtures of PDAGS/PMF had better structural compatibility and full miscibility with each other. PDAGS/PMF and PDAGS/SFO crystallised in β'+β polymorphs in the presence of 0.4-0.5% PDAGS while PDAGS/POL resulted in β polymorphs crystal. The results gave indication that PDAGS: PMF at 50%:50% and 60%:40% (w/w) were the most suitable fat blend to be used as bakery shortening.
Antioxidants have been widely used in the food industry to enhance product quality by preventing oxidation of susceptible substances. This work was carried out to maximise the recovery of total phenolic content (TPC), total flavonoid content (TFC), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical-scavenging capacity and 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging capacity from Morinda citrifolia fruit via modification of the ethanol concentration, extraction time and extraction temperature at minimal processing cost. The optimised conditions yielded values of 881.57 ± 17.74 mg GAE/100 g DW for TPC, 552.53 ± 34.16 mg CE/100 g DW for TFC, 799.20 ± 2.97 µmol TEAC/100 g DW for ABTS and 2,317.01 ± 18.13 µmol TEAC/100 g DW for DPPH were 75% ethanol, 40 min of time and 57 °C. The four responses did not differ significantly (p > 0.05) from predicted values, indicating that models obtained are suitable to the optimisation of extraction conditions for phenolics from M. citrifolia. The relative amounts of flavonoids were 0.784 ± 0.01 mg quercetin/g of extract and 1.021 ± 0.04 mg rutin/g of extract. On the basis of the results obtained, M. citrifolia extract can be used as a valuable bioactive source of natural antioxidants.
Fractionation which separates the olein (liquid) and stearin (solid) fractions of oil is used to modify the physicochemical properties of fats in order to extend its applications. Studies showed that the properties of fractionated end products can be affected by fractionation processing conditions. In the present study, dry fractionation of palm-based diacylglycerol (PDAG) was performed at different: cooling rates (0.05, 0.5, 1.0, 1.5, 2.0, 2.5 and 3.0°C/min), end-crystallisation temperatures (30, 35, 40, 45 and 50°C) and agitation speeds (30, 50, 70, 90 and 110 rpm) to determine the effect of these parameters on the properties and yield of the solid and liquid portions. To determine the physicochemical properties of olein and stearin fraction: Iodine value (IV), fatty acid composition (FAC), acylglycerol composition, slip melting point (SMP), solid fat content (SFC), thermal behaviour tests were carried out. Fractionation of PDAG fat changes the chemical composition of liquid and solid fractions. In terms of FAC, the major fatty acid in olein and stearin fractions were oleic (C18:1) and palmitic (C16:0) respectively. Acylglycerol composition showed that olein and stearin fractions is concentrated with TAG and DAG respectively. Crystallization temperature, cooling rate and agitation speed does not affect the IV, SFC, melting and cooling properties of the stearin fraction. The stearin fraction was only affected by cooling rate which changes its SMP. On the other hand, olein fraction was affected by crystallization temperature and cooling rate but not agitation speed which caused changes in IV, SMP, SFC, melting and crystallization behavior. Increase in both the crystallization temperature and cooling rate caused a reduction of IV, increment of the SFC, SMP, melting and crystallization behaviour of olein fraction and vice versa. The fractionated stearin part melted above 65°C while the olein melted at 40°C. SMP in olein fraction also reduced to a range of 26 to 44°C while SMP of stearin fractions increased to (60-62°C) compared to PDAG.