Method for synthesizing diacylglycerol in reverse micelle enzyme system

Abstract

The invention discloses a method for synthesizing diacylglycerol in a reverse micelle enzyme system using activation characteristics of a lipase interface. The specific synthesis method is as follows:first, constructing the reverse micelle enzyme system, specifically, adding an immobilized water-soluble lipase buffer aqueous solution (50 mmol / mL sodium dihydrogen phosphate-disodium hydrogen phosphate, pH 6.5 to 8) in an isooctane solution of sodium bis(2-ethylhexyl) sulfosuccinate (AOT), and conducting spot energy ultrasound to obtain a clarified reverse micelle enzyme system, wherein the ratio of an enzyme (mg / mL) to water (mmol / mL) to the AOT (mmol / mL) is equal to (3-8):(200-600):(20-30); secondly, mixing 0.5 to 0.8 g / (mL reaction system) plant oil and glycerin with a molar ratio of 1:(1-4), adding a formed mixture into the reverse micelle enzyme system, conducting a reaction for 6-8 h under the conditions that the enzyme load is 1.5-3% of the amount of the total substrate and the reaction temperature is 40 to 60 DEG C, after reacting, adding water with the amounting being 2 times the volume of the reaction liquid, conducting shaking for layering, conducting centrifugation at 4200 rpm for 5min, and taking an upper oil phase which has a diacylglycerol content of 50.45% to 63.23% measured by HPLC; and finally performing twice molecular distillation to purify a product which has a purity of 80.82% to 95.87% measured by HPLC. The method has the advantage of producing diacylglycerol with high efficiency and high yield.

Description

technical field [0001] The invention belongs to the field of grain and oil processing, and in particular relates to a method for synthesizing diglyceride in a reverse micellar enzyme system. Background technique [0002] Structural lipid is a kind of oil with special nutritional effect or physiological function obtained through structural recombination or modification of natural oil. Diacylglycerol is one of them. Concerned, the study found that after 40 healthy men participated in the intake of triglyceride oil or diglyceride oil, the levels of chylomicrons and serum triglycerides in the triglyceride group were significantly higher than those in the diglyceride group. [0003] The preparation of diglycerides is mainly realized under the action of catalysts. According to different catalysts, it can be roughly divided into chemical methods and enzymatic methods. According to different reaction mechanisms, it can also be divided into direct esterification methods, glycerol hyd...

AI Technical Summary

Problems solved by technology

[0006] The preparation method of the existing reverse micellar enzyme system is relatively simple. The surfactant solution and the enzyme solution are mixed and fully shaken until clear. However, the reverse micellar system prepared by this method has uneven droplet size and relatively low stability. Bad question

On the one hand, poor stability makes the system unusable for a long time; on the other hand, the relationship between the droplet size in reverse micelles and the size of lipase is one of the important factors that determine the activity of lipase in reverse micelles. Therefore, due to the inhomogeneity of the droplet size, the enzyme activity is not fully improved and the reproducibility of the results is poor.