Method for synthesizing phytosterol ester in reverse micelle enzyme system
A technology for phytosterol esters and phytosterols, which is applied in the field of synthesizing phytosterol esters, can solve the problems of uneven droplet size, unavailability of long-term utilization, incomplete enzymatic activity, etc., and achieves mild production conditions, easy operation and simple synthesis method. Effect
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Embodiment 1
[0020] Dissolve a certain amount of AOT in isooctane, and mix it thoroughly with a vortex oscillator until it becomes clear to obtain an AOT isooctane solution, and then add a certain amount of non-immobilized water-soluble Candidarugosa Lipase buffered aqueous solution (50 mol / mL PBS, pH7.5), where [enzyme] (mg / mL): [water] (mmol / mL): [AOT] = 3:375:25, and then use concentrated energy ultrasound For homogenization, the focused ultrasonic frequency was 25 kHz, the power was 300 W, the ultrasonic temperature was 45 °C, the total ultrasonic time was 42 s, and the ultrasonic mode was "ultrasonic 5 s-stop 2 s". Then 50 mol / L β-sitosterol and 3.5 times the molar mass of lauric acid were added, and the reaction was carried out for 6 h under the conditions of 1.6 % of the total substrate and a reaction temperature of 45 °C, supplemented by diffuse ultrasound. The ultrasonic power was 35 kHz, the power was 300 W, and the ultrasonic mode was "ultrasonic 10 s-stop 20 s". After the react...
Embodiment 2
[0022] Dissolve a certain amount of AOT in isooctane, and mix thoroughly with a vortex oscillator until it becomes clear to obtain an AOT isooctane solution, and then add a certain amount of non-immobilized water-soluble Candidarugosa Lipase buffered aqueous solution (50 mol / mL PBS, pH 7), where [enzyme] (mg / mL): [water] (mmol / mL): [AOT] (mmol / mL) = 4:500:25, then Focused ultrasound was used for homogenization, the frequency of focused ultrasound was 20 kHz, the power was 300 W, the ultrasonic temperature was 50 °C, the total ultrasonic time was 56 s, and the ultrasonic mode was "ultrasound 5 s-stop 2 s". Then add 80 mol / L β-sitosterol and 3 times the molar mass of palmitic acid, and react for 6 h under the condition that the enzyme load is 1.6% of the total substrate and the reaction temperature is 50 °C, supplemented by diffuse ultrasound during the process. The ultrasonic power was 30 kHz, the power was 350 W, and the ultrasonic mode was "ultrasonic 10 s-stop 20 s". After t...
Embodiment 3
[0024] Dissolve a certain amount of AOT in isooctane, and mix it thoroughly with a vortex oscillator until it becomes clear to obtain an AOT isooctane solution, and then add a certain amount of non-immobilized water-soluble Candidarugosa Lipase buffered aqueous solution (50 mol / mL PBS, pH7.5), where [enzyme] (mg / mL): [water] (mmol / mL): [AOT] (mmol / mL) = 8:450:25, Then, homogenization was performed by focused ultrasound. The frequency of focused ultrasound was 35 kHz, the power was 350 W, the ultrasound temperature was 40 °C, the total time of ultrasound was 35 s, and the ultrasound mode was "5 s of ultrasound - 2 s off". Then add 50 mol / L β-sitosterol and α-linoleic acid with twice the molar mass, and react for 4 h under the condition that the enzyme load is 1.5% of the total substrate and the reaction temperature is 40 °C. Using divergent ultrasound, the ultrasonic power was 35 kHz, the power was 300 W, and the ultrasonic mode was "10 s of ultrasonication - 20 s off". After t...
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