Method for synthesizing phytosterol ester in reverse micelle enzyme system

Abstract

The invention discloses a method for synthesizing phytosterol ester in a reverse micelle enzyme system based on lipase interface activation characteristics. The specific synthesis method comprises thefollowing steps: constructing a reverse micelle enzyme system, wherein a non-immobilized water-soluble lipase buffer aqueous solution (50 mmol/mL sodium dihydrogen phosphate-disodium hydrogen phosphate, pH 6.5-8) into an isooctane solution of 2-ethylhexyl succinate sodium sulfonate (AOT), a clarified reverse micelle enzyme system is obtained through energy-gathered ultrasound, and a ratio of [enzyme] (mg/mL) to [water] (mmol/mL) to [AOT] (mmol/mL) is (3-8):(200-600):(20-30); adding 50-100 mmol/mL phytosterol and C12-C18 fatty acid accounting for 2-4 times the molar mass of the phytosterol into the reverse micelle enzyme system, and carrying out a reaction for 3-6 hours at a temperature of 35-55 DEGC under an enzyme loading capacity accounting for 1.5-3% of the total substrate, wherein theprocess is assisted by divergent ultrasonic action, and the yield of the esterification product measured by an HPLC method is 85.79-92.82%; and purifying the product by a solvent crystallization method, wherein the purity is 98.56-99.43% measured by the HPLC method. The method has the advantage of high efficiency and high yield production of phytosterol ester.

Description

technical field [0001] The invention belongs to the field of grain and oil processing, and in particular relates to a method for synthesizing phytosterol esters in a reverse micellar enzyme system. Background technique [0002] Phytosterols are a class of triterpene natural products with a chemical structure similar to cholesterol. There are more than 100 specific types, of which the four most common are β-sitosterol, stigmasterol, brassicasterol and campesterol. As early as the 1950s , phytosterols have been proved by experiments on animals and humans to significantly reduce blood cholesterol, thereby reducing the risk of cardiovascular disease, but because of their poor oil solubility and water solubility, it is difficult to be directly ingested and utilized by the human body, so now Mainly by esterifying phytosterols to increase its solubility and increase bioavailability by 20-40%. [0003] The synthesis method of phytosterol ester can be divided into chemical synthesis...

AI Technical Summary

Problems solved by technology

[0007] 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.