• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.8
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• pp.1062-1068
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• 2009

Diacylglycerol (DAG) were synthesized by enzymatic esterification of glyceryl monooleate (GMO) and conjugated linoleic acid (CLA) in a shaking water bath. The reaction was catalyzed by Lipozyme TLIM (immobilized lipase from Thermomyces lanuginosa). Effects of reaction time, molar ratio, enzyme road and molecular sieves were studied. Results of normal-phase high performance liquid chromatography (NP-HPLC) analysis were performed. At 1:1, 2:1 and 3:1 (GMO : CLA) molar ratio and Lipozyme TLIM of 20% amount, DAG were produced in 42.6, 54.4 and 54.6 area% in 1 hr, respectively. When different Lipozyme TLIM amounts (2, 5, 10, 20%) were used with 2:1 (GMO : CLA) molar ratio, DAG were produced 21.4 (24 hr), 51.7 (12 hr), 56.2 (6 hr) and 54.4 (1 hr) area%, respectively. The reaction in the absence of molecular sieves increased DAG contents. The maximum DAG concentration conditions were obtained with molar ratio of 2:1 (GMO : CLA), lipase concentration of 10% (of substrate), 10% molecular sieves and reaction time of 6 hours at 55$^{\circ}C$. Under this reaction condition, produced DAG-rich oil was composed of 69 area% DAG, 7.9 area% TAG, 2 area% FFA, and 21.1 area% MAG.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.4
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• pp.479-485
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• 2009

The acidolysis was performed to produce structured lipid with palm mid fraction (PMF) and stearic acid for 7, 24, and 36 hr at $70^{\circ}C$. The reaction was catalyzed by lipozyme TLIM (immobilized lipase from Thermonyces lanuginosa, amount of 10% and 20% by weight of total substrates) in the shaking water bath. The reaction conditions for maximum incorporation of stearic acid on the structured lipid were obtained when molar ratio of PMF and stearic acid was 1:2; concentration of lipozyme TLIM was 20wt%; reaction temperature was $70^{\circ}C$; and reaction time was 36 hr. After reaction under this condition, incorporation of stearic acid in the structured lipid was obtained up to 36.3% while the major components of triacylglycerol were 1,2-dipalmitoyl-3-stearoylglycerol (PPS, 28.19 area%), 1-palmitoyl-2-oleoyl-3-stearoylglycerol (POS/PSO, 20.70 area%) and 1-palmitoyl-2,3-distearoylglycerol (PSS, 18.13 area%). However, the fatty acid composition at the sn-2 position suggested that the positional specificity of lipozyme TLIM was not observed due to the acyl migration.

• Korean Journal of Food Science and Technology
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• v.42 no.2
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• pp.246-249
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• 2010

Diacylglycerol (DAG) was produced from lipase by the catalyzed synthesis of soybean oil (SBO) and glyceryl monooleate (GMO) with Lipozyme TLIM (Thermomyces lanuginosa). Effects of reaction time, molar ratio and enzyme road were studied. When 2:1, 1:1 and 1:2 (SBO:GMO) molar ratios with 20% Lipozyme TLIM were applied in a 1-hr reaction, the concentrations of DAG produced were 17.8, 20.0 and 20.4 g/100 g oil, respectively. Different amounts (2, 5, 10 and 20%) of Lipozyme TLIM were used at a 1:2 (SBO:GMO) molar ratio, and the concentrations of DAG produced in a 1-hr reaction were 10.8, 14.0, 16.9 and 20.4 g/100 g oil, respectively. During a 72-hr reaction, 10.8-22.7 g/100 g oil of DAG were produced under the reaction conditions in this study.

• Korean Journal of Agricultural Science
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• v.40 no.3
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• pp.227-235
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• 2013

This study was conducted to prove the effect of irradiation on lipases (lipase AK, lipase AH, lipase PS-D, Lipozyme TLIM, Lipozyme RMIM and Novozyme SP435) which were used for interesterification reaction using batch type reactor. Through such interesterification, structured lipid (1(3)-palmitoyl-2-oleoyl-3(1)-stearoyl, POS) was synthesized by lipase treated with irradiation at different doses (0, 3, 7, 14, 29 and 59 kGy) using canola oil, palmitic ethyl ester (PEE) and stearic ethyl ester (StEE). After the reaction, fatty acid composition of triacylglycerol (TAG) in structured lipid was analyzed to compare the lipase activity. The results showed that activity of the irradiated lipase AH, PS-D and Novozyme SP435 with certain dose (3 kGy) were slightly improved. Such change of lipase activity suggested that irradiation might affect on the interesterification properties. Especially, Lipase AK, Lipozyme TLIM and Lipozyme RMIM after at 3 kGy irradiation showed that content of stearic acid ($C_{18:0}$) was increased while palmitic acid ($C_{16:0}$) decreased in the interesterified products.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.8
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• pp.1113-1120
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• 2011

Enzymatic interesterification was conducted with the palm mid fraction (PMF) and stearic ethyl ester for 1, 5, and 9 hr at 46$^{\circ}C$. The reaction was catalyzed by Lipozyme TLIM (2, 3, and 4% by weight of total substrates) in a shaking water bath at 180 rpm. As the reaction continued, oleic acid (C18:1) content at the sn-2 position decreased, whereas saturated fatty acid (C16:0 and C18:0) content increased. In the high performance chromatography analysis, 1,3-dipalmitoyl-2-oleoyl glycerol content decreased, whereas 1(3)-palmitoyl-2-oleoyl-3(1)-stearoyl glycerol (POS) content increased up to the reaction equilibrium. The rate of acyl migration increased with increasing molar ratio and enzyme load as well as reaction time. The optimal reaction conditions for maximizing POS content (53.5 area%) and minimizing acyl migration (23.1 area%) were obtained with a PMF : stearic ethyl ester=1:2 (molar ratio), Lipozyme TLIM 3 wt%, and a reaction time of 5 hr.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.9
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• pp.1320-1327
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• 2010

Lipase-catalyzed interesterification of canola (CO) and fully hydrogenated soybean oil (FHSBO) at different weight ratios (70:30, 75:25, and 80:20) was performed in a batch type reactor to produce low-trans solid fats. Each reaction was conducted in the shaking water bath for various reaction times (1, 3, 6, 18 and 24 hr) at 70oC and 220 rpm using Lipozyme TLIM (20 wt% of total substrate) from Thermomyces lanuginosus. After 24 hr reaction, solid fat content (SFC) by differential scanning calorimetry (DSC), fatty acid and triacylglycerol (TAG) composition of low-trans solid fats were determined. SFC of the products was reduced when the content of canola oil in the reaction mixture was increased. Major fatty acids were stearic acid (C18:0), oleic acid (C18:1) and linoleic acid (C18:2). Trans fatty acid content in the low-trans solid fats showed less than 0.3 wt%. In the HPLC analysis, major TAG species showed LOO (linoleyl-oleoyl-oleoyl), OOO, POO/SOL, SOO, and SOS.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.10
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• pp.1430-1437
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• 2011

Synthesis conditions of cocoa butter equivalents were optimized using the response surface method (RSM) by interesterification of canola oil (Ca), palmitic ethyl ester (PEE), and stearic ethyl ester (StEE). The reaction was catalyzed by immobilized lipase (Lipozyme TLIM) from Thermomyces lanuginosa to produce structured lipids containing a composition of triacylglycerols similar to cocoa butter. Reaction conditions were optimized using D-optimal design with the three reaction factors of the substrate molar ratio of canola oil to palmitic ethyl ester and stearic ethyl ester (Ca : PEE : StEE=1:1:3, 1:1.66:5, 1:2:6, 1:2.33:7, 1:3:9, $X_1$), enzyme ratio (2~6%, $X_2$), and reaction time (30~270 min, $X_3$). The optimal conditions that minimized acyl-migration while maximizing 1-palmitoyl-2-oleoyl-3-stearoyl glycerol (POS), 1,3-distearoyl-2-oleoyl glycerol (SOS), and 1,3-dipalmitoyl-2-oleoyl glycerol (POP) were predicted, resulting in Ca : PEE : StEE=1:3:9, 6% of enzyme ratio, and 40 min of reaction time. The reaction product of structured lipids was synthesized again under the same conditions, showing 10.43 area% of acyl-migration, 25.31 area% of POS/PSO, 19.79 area% of SOS, and 11.22 area% of POP.

• Korean Journal of Agricultural Science
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• v.45 no.2
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• pp.265-282
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• 2018

The purpose of this research was to produce symmetric (Saturated-Unsaturated-Saturated, SUS) triacylglycerol (TAG) using palm stearin fraction oil (PSFO) and high oleic sunflower oil (HOSO) as substrates to replace cocoa butter. PSFO was blended with HOSO (1 : 2 weight ratio), and $lipozyme^{(R)}$ TLIM (20 weight % of the substrate) was added. Interesterification was carried out in a shaking water bath at $55^{\circ}C$ at 220 rpm for 6 hours. The response surface methodology (RSM) through the central composite face design was employed to observe the optimized SUS-TAG. The independent factors were the reaction temperature ($X_1$: 65, 75 and $85^{\circ}C$), reaction time ($X_2$: 1, 3 and 5 hours) and ratio of TLIM ($X_3$: 10, 15 and 20 weight %). The dependent variables were $Y_1$ = Saturated-Unsaturated-Unsaturated (SUU, area %), $Y_2=SUS$ (area %), $Y_3$ = Saturated-Saturated-Unsaturated (SSU, area %), $Y_4$ = Unsaturated-Unsaturated-Unsaturated (UUU, area %), and $Y_5=sn-2$ unsaturated fatty acid (area %). The optimal conditions from the central composite face design minimized acyl migration while maximizing the presence of unsaturated fatty acid at the sn-2 position (73.43 area %). The optimal conditions were $X_1=65^{\circ}C$, $X_2=1hour$, and $X_3=20weight%$. As a result of the response surface analysis, the lack of fits was found as $Y_1=0.622$, $Y_2=0.438$, $Y_3=0.264$, $Y_4=0.526$, and $Y_5=0.215$, and their $R^2$ were 0.897, 0.944, 0.826, 0.857, and 0.867, respectively.

• Korean Journal of Agricultural Science
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• v.37 no.1
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• pp.69-72
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• 2010

This study explored the solid fat index (SFI) of structured lipids (SLs) synthesized by lipase-catalyzed (Lipozyme TLIM) interesterification using hydrogenated coconut oil (HCO), palm oil (PO) and palm stearin solid (PSS). SLs were produced using three blends of HCO/PO (60:40, w/w), HCO/PSS (40:60 and 60:40, w/w), and HCO/PO/PSS (32:48:18, w/w/w) to find a desirable confectionary fat by monitoring melting and crystallization behaviors of SFI of SLs using differential scanning calorimetry (DSC). SFI of HCO/PSS (60:40) and HCO/PO/PSS (32:48:18) at $25^{\circ}C$ were 70% and 68%, respectively. These results suggest that HCO/PSS (60:40) and HCO/PO/PSS (32:48:18) may be useful as potential SLs of a confectionary fat.

• Korean Journal of Agricultural Science
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• v.40 no.4
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• pp.367-375
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• 2013

The enzymatic interesterification was performed to produce structured lipids (SLs) with palm mid fraction (PMF) and stearic ethyl ester (STEE) for 1, 3, 6, 9, 12 and 15 hr at $80^{\circ}C$. The reaction was catalyzed by Lipozyme TLIM (immobilized lipase from Thermomyces lanuginosus, amount of 20% by weight of total substrates) in a shaking water bath set at 180 rpm. The optimum condition for synthesis of asymmetric SLs were: substrate molar ratio 1:0.5 (PMF:STEE, by weight), reaction time 6 hr, enzyme 20% (wt%, water activity=0.085) of total substrate and reaction temperature $80^{\circ}C$. After reaction at optimized condition, triacylglycerols (symmetrical and asymmetrical TAGs) from reactants were isolated. POP/PPO (1,3-palmitoyl-2-oleoyl glycerol or 1,2-palmitoyl-3-oleoyl glycerol), POS/PSO (palmitoyl-oleoyl-stearoyl glycerol or palmitoyl-stearoyl-oleoyl glycerol), SOS/SSO (1,3-stearoyl-2-oleoyl glycerol or 1,2-stearoyl-3-oleoyl glycerol) were obtained by solvent fractionation. Finally, refined SLs contained stearic acid of 16.91%. Solid fat index and thermogram of the refined SLs were obtained using differential scanning calorimetry. The degree of asymmetric triacylglycerol in the refined SLs was analyzed by Ag-HPLC equipped with evaporated light scattering detector (ELSD). The refined SLs consisted of symmetric TAG of 41.15 area% and asymmetric TAG of 58.85 area%.