阅读说明：本技术 一种制备甘油葡糖苷的方法 (Method for preparing glycerol glucoside ) 是由 郭欣 陈青来 于 2020-06-16 设计创作，主要内容包括：本发明公开了一种制备甘油葡糖苷的方法,包括如下步骤：将浓度为60～300g/L的蔗糖与浓度为20～150g/L的甘油在浓度为50～150g/L的复合生物酶及浓度为20～60mmol/L的柠檬酸三钠缓冲液中以3∶1∶2∶1的体积比,在45～55℃的条件下反应30～40小时。本发明的有益效果是：直接利用菌体细胞作为生物催化剂,避免了购买昂贵的商业用酶制剂或者进行复杂的菌体破壁处理,可大大减少生产用地及固定资产的投入,生产成本更低,制备工艺简单,且甘油葡糖苷的产量高。(The invention discloses a method for preparing glycerol glucoside, which comprises the following steps: reacting 60-300 g/L of sucrose and 20-150 g/L of glycerol in 50-150 g/L of compound bio-enzyme and 20-60 mmol/L of trisodium citrate buffer solution at a volume ratio of 3: 1: 2: 1 at 45-55 ℃ for 30-40 hours. The invention has the beneficial effects that: the bacterial cells are directly used as biocatalysts, thereby avoiding purchasing expensive commercial enzyme preparations or carrying out complex bacterial wall breaking treatment, greatly reducing the investment of production land and fixed assets, having lower production cost, simple preparation process and high output of the glycerol glucoside.)

1. A method of preparing a glycerol glucoside, comprising: the method comprises the following steps: reacting 60-300 g/L sucrose and 20-150 g/L glycerol in 50-150 g/L composite biological enzyme and 20-60 mmol/L trisodium citrate buffer solution at a volume ratio of 3: 1: 2: 1 at 45-55 ℃ for 30-40 hours, and filtering through an ultrafiltration membrane to remove the enzyme; purifying by ion exchange resin, concentrating by reduced pressure distillation, and filtering to remove solid impurities to obtain glycerol glucoside finished product; wherein the pH value of the trisodium citrate buffer solution is 4.5-5.5; the compound biological enzyme comprises sucrose phosphorylase and Escherichia coli.

2. The method for producing glycerol glucoside of claim 1, wherein said method comprises the steps of: the pH of the trisodium citrate buffer is 5.5.

3. The method for producing glycerol glucoside of claim 1, wherein said method comprises the steps of: the compound biological enzyme is thallus pulp containing sucrose phosphorylase.

4. The method for producing glycerol glucoside of claim 3, wherein said method comprises the steps of: before the addition reaction of the compound biological enzyme, the culture is carried out for 10 hours under the conditions of 35 ℃ and the rotating speed of 200 rpm.

5. The method for producing glycerol glucoside of claim 4, wherein said method comprises the steps of: adding a fermentation culture medium to perform fermentation culture before the shake culture of the compound biological enzyme; the fermentation medium comprises the following components in percentage by weight: 3.5 percent of yeast extract powder, 0.2 percent of peptone, 0.4 percent of sodium chloride, 0.3 percent of potassium dihydrogen phosphate, 0.6 percent of disodium hydrogen phosphate, 0.3 percent of sucrose, 0.2 percent of glycerol, 0.1 percent of glucose, 0.005 percent of calcium chloride and the balance of water.

Technical Field

The invention belongs to the field of a preparation process of 2-O-alpha-D-glycerol glucoside, and particularly relates to a method for preparing glycerol glucoside.

Background

Glycerol glucosides (also known as 2-glycerol glucosides) are ubiquitous in nature, particularly in salt-tolerant cyanobacteria such as blue-green algae and Artocarpus heterophyllus (also known as Neurospora, Potentilla repens). The active substance is the most important active substance for the millettia capable of living and reactivating and regenerating in extreme environment, and can firmly lock the precious last water drop in the body, so that the glycerol glucoside has extremely strong physiological effects of moistening, locking and preserving moisture, can be used as a functional raw material of cosmetics, and has great market demand. In addition, glycerol glucoside has also been added to foods as a non-cariogenic sweetener, and Japanese Takenaka F. et al identified 3 different configurations of glycerol glucosides in Japanese sake and achieved chemical synthesis of these 3 substances by chemical synthesis (Biosci Biotechnol Biochem, 2000, 64: 378-385).

The current methods for preparing glycerol glucosides mainly include chemical synthesis and enzymatic synthesis. Plant extraction is limited by site, source and the like, and large-scale production cannot be realized; the chemical synthesis method has the defects of high pollution, high energy consumption, complex preparation process and the like, and has the problem of more impurities (1-glycerol glucoside is introduced in the synthesis process, and the water-locking and moisture-preserving functions of 2-glycerol glucoside are the best) so that the synthesis method mainly adopts a biological enzyme method with high specificity at present. Sucrose and glycerol are used as raw materials, and 2-glycerol glucoside is produced under the action of sucrose phosphorylase. However, the existing glycerol glucoside preparation method has the defect of high preparation cost, particularly the preparation method of the 2-O-alpha-D-glycerol glucoside has the problems of high preparation cost and low yield, and the 2-O-alpha-D-glycerol glucoside not only has the moisturizing function, but also has the repairing function, so that the application range is wider.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provides a method for preparing glycerol glucoside, which comprises the following steps: reacting 60-300 g/L sucrose and 20-150 g/L glycerol in 50-150 g/L composite biological enzyme and 20-60 mmol/L trisodium citrate buffer solution at a volume ratio of 3: 1: 2: 1 at 45-55 ℃ for 30-40 hours, and filtering through an ultrafiltration membrane to remove the enzyme; purifying by ion exchange resin, concentrating by reduced pressure distillation, and filtering to remove solid impurities to obtain glycerol glucoside finished product; wherein the pH value of the trisodium citrate buffer solution is 4.5-5.5; the compound biological enzyme comprises sucrose phosphorylase and Escherichia coli.

Further, the pH of the trisodium citrate buffer is 5.5.

Further, the compound biological enzyme is thallus pulp containing sucrose phosphorylase.

Further, the complex biological enzyme is added to the reaction solution and is cultured for 10 hours under the conditions of 35 ℃ and 200 rpm.

Further, adding a fermentation culture medium for fermentation culture before the shake culture of the compound biological enzyme; the fermentation medium comprises the following components in percentage by weight: 3.5 percent of yeast extract powder, 0.2 percent of peptone, 0.4 percent of sodium chloride, 0.3 percent of potassium dihydrogen phosphate, 0.6 percent of disodium hydrogen phosphate, 0.3 percent of sucrose, 0.2 percent of glycerol, 0.1 percent of glucose, 0.005 percent of calcium chloride and the balance of water.

The invention has the beneficial effects that: the bacterial cells are directly used as biocatalysts, thereby avoiding purchasing expensive commercial enzyme preparations or carrying out complex bacterial wall breaking treatment, greatly reducing the investment of production land and fixed assets, having lower production cost, simple preparation process and high output of the glycerol glucoside.

Detailed Description

The invention will now be further described with reference to the following examples, which are intended to better illustrate the invention and are not intended to limit the invention thereto.