Kinetic behavior of sunflower oil lipase-catalyzed acidolysis

CONSUELO PACHECO; AMALIA CARELLI; MARÍA E. CARRÍN

Seattle, USA

Congreso; 99th AOCS Annual Meeting & Expo; 2008

AOCS

Kinetic behavior of sunflower oil lipase-catalyzed acidolysis Consuelo Pacheco, Amalia A. Carelli, María E. Carrín PLAPIQUI (Universidad Nacional del Sur-CONICET), Bahía Blanca, Argentina During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was filtered and washed with hot hexane. Changes in SL composition were determined through deacidification by alkaline extraction, FAME preparation by cold transesterification with methanolic KOH, and further analysis by capillary gas chromatography (CGC). Deacidified products were analyzed to quantify by-products as glycerol (G), monoglycerides (MG) and diglycerides (DG) by silylation and CGC determination. Nonlinear regression methods were employed to determine kinetic parameters of the enzymatic mechanism representing the reaction from the initial stage until equilibrium. The effects of the reaction conditions (temperature, time, incorporated water) on enzymatic acidolysis were studied. DG formation and kinetic and equilibrium parameters showed temperature dependencies. MG formation was not observed. During enzymatic acidolysis a desired acyl group is incorporated onto a specific position of the triacylglycerol to produce structured lipids (SL). In the present study, sunflower oil (SO) was modified with a palmitic-stearic acids blend (FFA) by using an immobilized sn-1,3 specific lipase: Lipozyme RM IM (from Rhizomucor miehei, immobilized on ion-exchange resin). Experiments were performed as follow: substrates and solvents were preheated in a water bath to the required temperature; adding lipase started the reaction, which was carried out at a laboratory scale in a close system with agitation; to stop the reaction, mixture was