Method for preparing rebaudioside by use of stevioside

A technology of steviolbioside and stevioside, which is applied in the direction of fixing on/in organic carrier, fermentation, etc., can solve the problems of short service life and long reaction time of β-glucosidase, and achieve high economic value and high Selectivity, effect of improving thermal stability

Inactive Publication Date: 2013-02-13
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But, the reaction time of these prior art preparation steviolbioside needs 5 days, and its reaction time is too long, and after adopting the beta-glucosidase immobilized

Method used

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  • Method for preparing rebaudioside by use of stevioside
  • Method for preparing rebaudioside by use of stevioside

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] Embodiment 1: Preparation of steviolbioside

[0081] The implementation steps of this embodiment are as follows:

[0082] Firstly, immobilized β-glucosidase is prepared.

[0083] A. Preparation of transparent gel

[0084] According to the ratio of the weight of chitosan in g to the volume of acetic acid aqueous solution in mL of 0.3:20, chitosan was dissolved in 2% by weight of acetic acid aqueous solution, and then the bubbles were removed by ultrasonic waves to obtain a transparent chitosan gel.

[0085] B. Preparation of chitosan microspheres

[0086]Add the transparent chitosan gel obtained in step A into a coagulation solution, then harden at a temperature of 4°C for 14 hours, then wash with deionized water until neutral, and store it overnight at a temperature of 4°C to obtain a chitosan microspheres.

[0087] The coagulation solution is composed of 10% by weight NaOH aqueous solution and 95% by weight ethanol aqueous solution according to the volume ratio of ...

Embodiment 2

[0095] Embodiment 2: Preparation of steviolbioside

[0096] The implementation steps of this embodiment are as follows:

[0097] Firstly, immobilized β-glucosidase is prepared.

[0098] A. Preparation of transparent gel

[0099] According to the ratio of the weight of chitosan microspheres in g to the volume of glutaraldehyde aqueous solution in mL 0.2:24, chitosan was dissolved in 5% by weight acetic acid aqueous solution, and then the bubbles were removed by ultrasonic method to obtain a transparent shell Glycan Gel.

[0100] B. Preparation of chitosan microspheres

[0101] Add the transparent chitosan gel obtained in step A into a coagulation solution, then harden at a temperature of 2°C for 16 hours, then wash with deionized water until neutral, and store it overnight at a temperature of 3°C to obtain a chitosan microspheres.

[0102] The coagulation solution is composed of 8% by weight NaOH aqueous solution and 98% by weight ethanol aqueous solution according to the ...

Embodiment 3

[0107] Embodiment 3: Preparation of steviolbioside

[0108] The implementation steps of this embodiment are as follows:

[0109] Firstly, immobilized β-glucosidase is prepared.

[0110] A. Preparation of transparent gel

[0111] According to the ratio of the weight of chitosan microspheres in g to the volume of glutaraldehyde aqueous solution in mL of 0.5:16, chitosan was dissolved in 1% by weight acetic acid aqueous solution, and then the bubbles were removed by ultrasonic method to obtain a transparent shell. Glycan Gel.

[0112] B. Preparation of chitosan microspheres

[0113] Add the transparent chitosan gel obtained in step A into a coagulation solution, then harden at a temperature of 6°C for 10 hours, then wash with deionized water until neutral, and store it overnight at a temperature of 3°C to obtain a chitosan microspheres.

[0114] The coagulation solution is composed of 12% by weight NaOH aqueous solution and 92% by weight ethanol aqueous solution according to...

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Abstract

The invention relates to a method for preparing rebaudioside through packed bed enzymatic hydrolysis of stevioside. The method comprises the following steps of: dissolving the stevioside or stevia rebaudiana extract in water or phosphate buffer solution; combining the parts containing rebaudioside of the solution through an immobilization beta-glucosidase packed column; and concentrating and recrystallizing to obtain the rebaudioside. The preparation of rebaudioside takes 5 days in the prior art, and the transformation activity of the beta-glucosidase repeatedly used for 4 times is only 57.72% of the original activity. Through the invention, the preparation of rebaudioside using the immobilization beta-glucosidase only takes 12 hours, and the transformation rate of stevioside after 5 times of use still exceeds 84%. Thus, the method provided by the invention is short in production period, high in production efficiency and easy to realize continuous production, and has very broad application prospects.

Description

【Technical field】 [0001] The invention belongs to the technical field of organic compound biosynthesis. More specifically, the present invention relates to a method for preparing steviolbioside by using a packed bed to enzymatically hydrolyze stevioside. 【Background technique】 [0002] Stevioside is a mixture of various diterpene glycosides extracted from the leaves of Stevia rebaudiana. It is a natural and powerful sweetener. A, RA), in which rebaudioside A has a pure sweet taste, while stevioside has a certain after-bitter taste. At present, a large number of studies mainly use enzymatic transglycosylation reaction to change the structure of stevioside to improve its sweetness characteristics. In addition, studies have shown that enzymatic hydrolysis can convert stevioside into stevioside with poor sweetness but certain physiological activity, and stevioside can be further derivatized as an intermediate product into other substances with better physiological activity. ...

Claims

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Application Information

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IPC IPC(8): C12P19/56C12N11/10C12N11/04
Inventor 夏咏梅丁力方云
Owner JIANGNAN UNIV
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