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Method for preparing glucosyl group-alpha-cyclodextrin

A glucose-based and cyclodextrin technology, applied in the direction of fermentation, can solve problems such as increased branching rate, and achieve the effect of effective separation

Active Publication Date: 2017-07-21
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method involves two steps: pullulanase condenses maltose and α-CD into G 2 -α-CD and G 2 -α-CD is hydrolyzed under the action of glucoamylase, because glucoamylase converts G 2 -α-CD is hydrolyzed to G 1 - α-CD, the equilibrium of the pullulanase condensation reaction is biased towards the synthesis of maltosyl α-CD, which results in an increased branching rate

Method used

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  • Method for preparing glucosyl group-alpha-cyclodextrin
  • Method for preparing glucosyl group-alpha-cyclodextrin

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Effect test

Embodiment 1

[0026] A preparation method of glucosyl-alpha-cyclodextrin, said method comprising the following specific steps:

[0027] (1) Take 10 mg of maltodextrin in a 2 mL centrifuge tube, add 1 mL of pH 5.5 (20 mM) phosphate buffer solution, add 0.36 U of CGTase, and react at 60 ° C for 12 h;

[0028] (2) Insulate for 15 minutes in a water bath at 100°C;

[0029] (3) Separating and purifying the product obtained in step (2) by using a high performance liquid phase method to finally obtain 0.19 mg of glucosyl-α-cyclodextrin.

Embodiment 2

[0031] A preparation method of glucosyl-alpha-cyclodextrin, said method comprising the following specific steps:

[0032] (1) Take 10 mg of maltodextrin in a 2 mL centrifuge tube, add 1 mL of pH 5.5 (20 mM) phosphate buffer solution, add 0.36 U of CGTase, and react at 60 ° C for 12 h;

[0033] (2) Add 0.72U glucoamylase to the reaction system, react for 12 hours under the reaction conditions of 45° C., and incubate at 100° C. for 15 minutes;

[0034] (3) Separating and purifying the product obtained in step (2) by using a high performance liquid phase method to finally obtain 0 mg of glucosyl-α-cyclodextrin.

Embodiment 3

[0036] A preparation method of glucosyl-alpha-cyclodextrin, said method comprising the following specific steps:

[0037] (1) Take 10 mg of maltodextrin in a 2 mL centrifuge tube, add 1 mL of pH 6.0 (20 mM) phosphate buffer solution, add 0.1 U of CGTase, and react at 60 ° C for 12 h;

[0038] (2) Heat in a water bath at 80°C for 25 minutes to make the enzyme activity of CGTase reach 0.001U;

[0039] (3) Add 0.5U glucoamylase to the reaction system, react for 12 hours under the reaction conditions of 45° C., and incubate at 100° C. for 15 minutes to prepare a mixture containing glucosyl-α-cyclodextrin;

[0040] (4) Separating and purifying the product obtained in step (3) by using a high performance liquid phase method to finally obtain 0.23 mg of glucosyl-α-cyclodextrin.

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Abstract

The invention discloses a method for preparing glucosyl group-alpha-cyclodextrin. The method comprises the following steps: (1) dissolving maltodextrin into a phosphate buffer solution, adding CGTase (Cyclodextrin Glycosyl Transferase), and performing reaction for 12-24 hours; (2) reducing the enzyme activity to 0.001-1.0U by using a physical method; (3) adding a saccharifying enzyme into a reaction system of the step (2), performing reaction for 12-24 hours, and performing high-temperature enzyme deactivation so as to obtain a mixture with glucosyl group-alpha-cyclodextrin; (4) performing separation and purification on the mixture obtained in the step (3) by using a high performance liquid method, thereby finally obtaining the glucosyl group-alpha-cyclodextrin. The method disclosed by the invention is mild in reaction condition, cyclodextrine and unreacted substrates are hydrolyzed by virtue of the slight coupling activity of the CGTase, and thus high-purity glucosyl group-alpha-cyclodextrin is prepared.

Description

technical field [0001] The invention relates to the technical field of cyclodextrin synthesis, in particular to a method for preparing glucosyl-alpha-cyclodextrin through double-enzyme compound hydrolysis. Background technique [0002] Cyclodextrin, also known as cyclomaltooligosaccharide, is a general term for a class of oligosaccharides that are linked by several α-D glucopyranose units through α-1,4 glycosidic bonds. The application of cyclodextrin is severely limited due to its poor water solubility, high hemolysis and nephrotoxicity, so the modification of cyclodextrin is very necessary. [0003] The modification methods of cyclodextrin include chemical method or biological enzyme method to modify cyclodextrin, grafting some specific substituents to cyclodextrin, which can keep the cavity structure of cyclodextrin basically unchanged. The modified product obtained by introducing a substituent group under normal circumstances is usually called a modified cyclodextrin, o...

Claims

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Application Information

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IPC IPC(8): C12P19/20C12P19/18C12P19/04
CPCC12P19/04C12P19/18C12P19/20
Inventor 金征宇柏玉香夏柳溪吴云徐学明谢正军王金鹏周星
Owner JIANGNAN UNIV
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