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

• Bulletin of the Korean Chemical Society
• /
• v.23 no.1
• /
• pp.9-10
• /
• 2002

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.40 no.10
• /
• pp.1430-1437
• /
• 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
• /
• v.40 no.4
• /
• pp.367-375
• /
• 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%.

• Food Science and Preservation
• /
• v.18 no.5
• /
• pp.721-728
• /
• 2011

1(3)-palmitoyl-2-oleoyl-3(1)-stearoyl-(POS)-glycerol-enriched reaction products were synthesized from camellia oil, palmitic ethyl ester, and stearic ethyl ester via lipase-catalyzed interesterification. Response surface methodology (RSM) was employed to optimize the production of the POS-enriched reaction product (Y1, %) and the stearicand palmitic-acid contents at the sn-2 position due to acyl migration (Y2, %). The reaction factors were the enzyme amount (X1, 2-6%), reaction time (X2, 60-360 min), and substrate molar ratio of camellia oil to palmitic ethyl ester and stearic ethyl ester (X3, 1-3 mol). The predictive models for Y1 and Y2 were adequate and reproducible as no lack of fit was signified (0.128 and 0.237) and as there were satisfactory levels of R2 (0.968 and 0.990, respectively). The optimal conditions for the reaction product for maximizing Y1 while minimizing Y2 were predicted at the reaction combination of 5.86% enzyme amount, 60 min reaction time, and 1:3 substrate molar ratio (3 moles of palmitic ethyl ester and 3 moles of stearic ethyl ester). Actual reaction was performed under the same conditions as above, and the resulting product contained 20.19% TAG-P/O/S and 12.71% saturated fatty acids at the sn-2 position.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.38 no.4
• /
• pp.479-485
• /
• 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 Agricultural Science
• /
• v.45 no.2
• /
• pp.265-282
• /
• 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.

• Applied Chemistry for Engineering
• /
• v.5 no.2
• /
• pp.209-214
• /
• 1994

Mono alkyl glycerides have bean obtained in good yield by enzyme catalyst from soybean oil. The reaction was carried out in an oil rich microemulsion formula. Best results were obtained with sodium bis(2-ethyl hexyl) sulfo succinate(AOT), isooctane as hydrocarbon component and buffer of pH 7. The enzyme used was a 1,3-specific lipase which leaves the 2-position intact. However, the 2-monoglyceride formed slowly undergoes long chain acyl migration to 1-mono-glyceride. Optimal reaction time at $35^{\circ}C$ reaction temperature was found to be three hour.

• Journal of the Korean Applied Science and Technology
• /
• v.15 no.4
• /
• pp.35-44
• /
• 1998

All the triacylglycerols including the molecular species having ${\Delta}^5$-unsaturated fatty acids from the seeds of Pinus Koraiensis, were split into a mixture of diacylglycerols by a Grignard reagent prepared with allyl bromide without arousing acyl chains of a glycerol moiety to migration, and were also easily partially hydrolyzed to diacylglycerols by pancreatic lipase. (S)-(+)-(1-naphthyl)ethyl urethane(NEU) derivatives of the diacylglycerol mixture derived from the triacylglycerols were fractionated into sn-1, 3-, sn-1, 2- and sn-2, 3-DG-NEU by silica-HPLC and the fatty acid composition of these fractions was analysed. $C_{18:1{\omega}9}$ is distributed evenly in the three positions of TG with $C_{18:2{\omega}6}$ mainly located in sn-2 position, while ${\Delta}^5$-unsaturated fatty acids such as ${\Delta}^{5.9}-C_{18:2}$, ${\Delta}^{5.9.12}-C_{18:3}$ and ${\Delta}^{5.11.14}-C_{20:3}$ are exclusively present in the sn-3 position. These results could be confirmed by $^{13}C$-NMR spectroscopy : the signals at $^{\delta}$173.231 ppm and $^{\delta}$172.811 ppm of the carbonyl carbon of acyl moieties indicate the presence of saturated acids and/or $C_{18:1{\omega}9}$ (oleic acid) in the ${\alpha}({\alpha}')$- or ${\beta}$- positions, and $C_{18:2{\omega}6}$ including $C_{18:1{\omega}9}$ in the ${\beta}$-position, respectively. In addition, the resonance at $^{\delta}$173.044 ppm suggested a location of ${\Delta}^5$-unsaturated fatty acid moiety in the ${\alpha}({\alpha}')$-position.

• Journal of Microbiology and Biotechnology
• /
• v.9 no.5
• /
• pp.624-630
• /
• 1999

In order to investigate the position of various fatty acids attached to glycerol and the specificity of Lipolase-100T, hydrolysis of fish oil was carried out with Lipolase-100T derived from Aspergillus oryzae. The amounts of free fatty acids produced from triglyceride, 1,2(2,3)-diglyceride, 1,3-diglyceride, and 2-monoglyceride and conversion rates of 1,2(2,3)-diglyceride to 1,3-diglyceride and 2-monoglyceride to 1(3)-monoglyceride were also calculated. The ratio of 1,2-diglyceride content to 1,3-diglyceride was higher than 70 in the early period of hydrolysis. The fatty acid content of the glyceride mixture after 72 h of hydrolysis was compared with that of fish oil, and it was found that polyunsaturated fatty acids such as C16:4, C20:4 n-3, C20:5 n-3, C21:5 n-3, C22:5 n-3 and C22:6 n-3 were located in the 2-position of glycerol. Material balance of each component in the hydrolysis system was written to obtain a set of simultaneous linear equations. The theoretical quantity of free fatty acids produced from triglyceride, 1,2-diglyceride, 1,3-diglyceride, and monoglyceride, respectively, were calculated by solving the linear equation system. The conversion rate of 1,2(2,3)-diglyceride to 1,3-diglyceride and that of 2-monoglyceride to 1(3)-monoglyceride were also obtained. The results showed that the migration rate of 1,2(2,3)-diglyceride to 1,3-diglyceride was higher than the hydrolysis rate of 1,2(2,3)-diglyceride to 2-monoglyceride and the conversion rate of 2-monoglyceride to 1(3)-monoglyceride was extremely low.