Crystallization method capable of increasing rebaudioside A1G content as well as product and application thereof

A technology for rebaudioside and crystallization products, which is applied in the field of crystallization to increase the content of rebaudioside A1G in rebaudioside A enzyme-modified products, and can solve the problems of difficult separation and purification by conventional methods, uneven taste, and product composition complex issues

Pending Publication Date: 2018-11-02
DONGTAI HIRYE BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The product of enzymatic modification of rebaudioside A is usually a mixture of glycosylation products with different sites and different amounts, which leads to problems such as complex product components and uneven taste
Moreover, since the components in the product mixture have the same parent nucleus, the molecular structure and polarity are very similar, it is difficult to separate and purify by conventional means

Method used

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  • Crystallization method capable of increasing rebaudioside A1G content as well as product and application thereof
  • Crystallization method capable of increasing rebaudioside A1G content as well as product and application thereof
  • Crystallization method capable of increasing rebaudioside A1G content as well as product and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0184] Example 1: Preparation and Identification of Rebaudioside A1G, a Rebaudioside A Double Enzyme Modified Product

[0185] according to figure 1 In the brief process shown, the modified product of rebaudioside A is prepared by a double-enzyme method. in particular:

[0186] Add 100L of purified water to the reaction kettle (Wuxi Hongqi Pressure Vessel Manufacturing Co., Ltd., 500KG emulsification pot, equipment code 21701098220170001). Weigh 6Kg rebaudioside A (Haotian Pharmaceutical Co., Ltd., RA97) and 6Kg β-cyclodextrin (Qufu Tianli Pharmaceutical Excipients Co., Ltd., 170805), put them into the reaction kettle, and heat to dissolve. Cyclodextrin glycosyltransferase (purchased from Novozymes (China) Biotechnology Co., Ltd., Toruzyme 3.0L, ACN00216, 3kNU / mL) 600kNU required for the first step of enzyme modification was added. The feed liquid temperature was maintained at 60°C, the stirring speed was 30 rpm / min, and the reaction was carried out for 24 hours. Boil at...

Embodiment 2

[0194] Example 2. The first crystallization of rebaudioside A enzyme modified product

[0195] Take 100 L of the rebaudioside A enzyme-modified product (i.e. the reaction system after boiling to terminate the reaction in Example 1), and separate it with a precision filter plate with a pore size of 5-10 μm (Shenyang Great Wall Filter Paperboard Co., Ltd., product number 1001), Pass through 100L of macroporous resin (Lanxiao Technology New Material Co., Ltd., LX-28; the new resin needs to be pretreated as follows: wash with 200L 85% ethanol solution, and then wash with purified water until the effluent has no alcohol smell, flow rate 100L / h) Adsorbed for 2 hours. The resin adsorbed with the sample was first washed with 300L of pure water to remove small molecules such as glucose mixed in the product, and then eluted with 200L of 60% (v / v) ethanol, and the eluate was collected, -0.07MPa, 75°C concentrate. Concentrate to a solid content of 50%, and then dry it in a spray dryer...

Embodiment 3

[0199] Example 3. The second crystallization of the rebaudioside A enzyme modified product

[0200] Take 7 Kg of the wet product of the first crystallization (ie the undried solid phase obtained in Example 2) and dissolve it by heating in 15 L, 65% (V / V) methanol aqueous solution. The temperature of the solution water bath was controlled at 20° C., the stirring speed was 15 rpm, and the crystallization time was 15 hours. The crystallization mixture was suction filtered with a Buchner funnel to obtain the second crystallization solid phase and liquid phase.

[0201] Gained solid phase adds ultrapure water after dissolving, detects with HPLC liquid phase (detection condition is as described in Example 1) (spectrum is as follows Figure 6 shown), the measured content of the new product rebaudioside A1G is 72.55%, and the wet weight is 5Kg (Table 3).

[0202] Add 2.5 L of purified water to the wet product of the second crystallization solid phase, dissolve it, and spray dry it...

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Abstract

The invention relates to a crystallization method capable of increasing rebaudioside A1G content as well as a product and application thereof. Stevioside rebaudioside A(RA) derivative product RA1G ispurified by a multi-time crystallization method (such as twice and three times), and high-purity RA1G with improved taste is obtained. Furthermore, crystallization liquid phase can be recycled into further production, so that the production cost is saved. The method is simple to operate, green and environmentally friendly in production process, low in cost, short in cycle and high in conversion efficiency; and the obtained product has good taste and has important application value in the industries of food, beverage and the like.

Description

technical field [0001] The invention belongs to the technical fields of biotechnology and food chemical industry. Specifically, the present invention relates to a crystallization method for increasing the content of rebaudioside A1G in rebaudioside A enzyme-modified products and its products and uses. Background technique [0002] Steviol glycosides are a series of glycosides extracted from the leaves of the Compositae herb Stevia rebaudiana Bertoni, among which the main content of natural products is stevioside (Stevioside, abbreviated as Stv) and rebaudioside A (Rebaudioside A, abbreviated as RA, also known as steviolbioside A). [0003] The molecular structure of rebaudioside A is shown below. It is a tetracyclic diterpene glycoside substance with 20 carbon atoms. Glucosyl formed by: [0004] [0005] Studies have shown that the sweetness of rebaudioside A is 200-300 times that of sucrose, and its calories are only 1 / 300 of that of sucrose. It is very stable to acid...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H15/256C07H1/06A23L2/60A23L27/30A23L33/125A61K47/26A61K8/60A61Q11/00
CPCA23L2/60A61K8/602A61K47/26A61Q11/00A23L27/36A23L33/125C07H1/06C07H15/256
Inventor 朱理平杜国营岳文艳肖敏徐莉王文正彭鹏
Owner DONGTAI HIRYE BIOTECH CO LTD
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