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Preparation method, product and application of Rebaudioside E

A technology of reaction and stevioside, applied in the field of preparation of rebaudioside E, can solve the problems of high cost, limited feasibility, cumbersome process steps, etc., and achieve the effect of increasing yield and simple and efficient preparation method

Pending Publication Date: 2020-01-07
BINZHOU SANYUAN BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The current bioenzymatic synthesis of rebaudioside E usually requires the addition of expensive UDP-glucose as a substrate, and through the action of UDP-glucosyltransferase, stevioside or rebaudioside A is used as a substrate to catalyze the formation of rebaudioside E. Baudioside E, due to the extremely high selling price of UDP-glucose, almost completely limits the feasibility of industrialized preparation of rebaudioside E, with poor economic efficiency and lack of market competitiveness
Or the process steps are cumbersome, the cost is high, and it lacks market competitiveness

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Prepare an aqueous phase reaction system, the initial reaction concentration of stevioside is 20g / L, the initial reaction concentration of ethyl acetate is 20g / L, the initial reaction concentration of β-cyclodextrin is 60g / L, the initial reaction concentration of palmitoyltransferase The initial reaction concentration of β-cyclodextrin glycosyltransferase was 58000U / L, 27000U / L, and the reaction system was adjusted to pH=7 by using phosphate buffer.

[0041] The aqueous phase reaction system was placed in a constant temperature shaker, the reaction temperature was 30° C., and the reaction time was 24 hours. The aqueous solution containing rebaudioside E can be prepared, and the concentration of rebaudioside E is determined to be 14.7g / L by high performance liquid chromatography, and the conversion rate is 73.5%.

Embodiment 2

[0043] Prepare an aqueous phase reaction system, the initial reaction concentration of stevioside is 100g / L, the initial reaction concentration of acetyl ethyl ester is 50g / L, the initial reaction concentration of β-cyclodextrin is 300g / L, the initial reaction concentration of palmitoyltransferase The initial reaction concentration of β-cyclodextrin glycosyltransferase was 80000U / L, 45000U / L, and the pH of the reaction system was adjusted to 6 with phosphate buffer.

[0044] The aqueous phase reaction system was placed in a constant temperature shaker, the reaction temperature was 40° C., and the reaction time was 16 hours. The aqueous solution containing rebaudioside E can be obtained, and the concentration of rebaudioside E is determined to be 61.7g / L, and the conversion rate is 61.7%.

Embodiment 3

[0046] Compared with Example 1, the difference is only that "the initial reaction concentration of ethyl acetate is 6g / L", and other conditions are the same as in Example 1. Utilize the method of high performance liquid chromatography, measure wherein the concentration of rebaudioside E is 2.6g / L, conversion rate is 13%.

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PUM

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Abstract

The invention provides a preparation method, product and application of Rebaudioside E. The preparation method comprises the steps that steviol glycosides is taken as a raw material, ethyl acetate andbeta-cyclodextrin are taken as glycosyl donors, a conversion reaction takes place under the catalysis of a complex enzyme, and the Rebaudioside E is generated. According to the preparation method, the product and application of the Rebaudioside E, the cheap and easily obtained ethyl acetate and beta-cyclodextrin are taken as the glycosyl donors to replace high-cost UDP-glucose to generate the Rebaudioside E under the catalysis of the complex enzyme. The reaction opens a new way for the preparation of the Rebaudioside E, the product yield of the reaction is high, the preparation process is simple, and the reaction is applicable to industrial production and has broad application prospects.

Description

technical field [0001] The invention belongs to the field of biocatalytic conversion, and relates to a preparation method and application of rebaudioside E, in particular to a preparation of rebaudioside E with ethyl acetate and β-cyclodextrin as glycosyl donors method and its application. Background technique [0002] As the sweetest sweetener known so far, stevioside has a sweetness 250-450 times that of sucrose, and its calories are only 1 / 300 of sucrose, with a slight astringent taste. In addition, stevioside also has auxiliary pharmacological effects on hypertension, obesity, diabetes, etc. Therefore, it has attracted more and more attention. Stevioside is the general term for steviosides. Currently, more than 35 kinds of steviosides have been isolated and identified from Stevia rebaudiana, including stevioside and rebaudioside A, B, C, D, E, M, etc. The steviol glycoside structure revolves around a diterpene steviol as the backbone, and the C13-position hydroxyl grou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P33/20
CPCC12P33/20
Inventor 李丽聂在建程保华李德春
Owner BINZHOU SANYUAN BIOLOGICAL TECH
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