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Method for purifying gamma-cyclodextrin by utilizing cyclodextrin hydrolase

A technology of cyclodextrin and hydrolase, which is applied in the field of cyclodextrin processing, can solve the problems of flammability, toxicity, unfavorable molecule recovery, etc., and achieve the effects of simple operation, improved purity, and good application potential

Active Publication Date: 2019-03-01
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this solvent method is also relatively expensive, flammable and toxic
Moreover, the high boiling point of the solvent is not conducive to the recovery of the formed molecules

Method used

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  • Method for purifying gamma-cyclodextrin by utilizing cyclodextrin hydrolase

Examples

Experimental program
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Effect test

Embodiment 1

[0019] Prepare 5 mL of 2% (w / v) DE13-17 maltodextrin solution with 20 mM, pH=6.0 phosphate buffer, add 0.8 U of CGTase, react at 60°C for 8 h, and inactivate with boiling water for 30 min. Using the above reaction solution as a substrate, add 0.5U cyclodextrin hydrolase, react at 80°C for 1 hour, and inactivate with boiling water for 1.5 hours after the reaction.

Embodiment 2

[0021] Use 10 mL of 50 mM, pH=6.5 10 % (w / v) soluble starch solution, pregelatinize, add 5 U of CGTase, react at 50 °C for 16 h, and inactivate with boiling water for 30 min. Using the above reaction solution as a substrate, add 2.1U cyclodextrin hydrolase and react at 70°C for 4h. After the reaction, inactivate with 5mL of 0.4M NaOH and neutralize with 0.4M hydrochloric acid solution.

Embodiment 3

[0023] Use 50mM, pH=7.5 5% (w / v) soluble starch solution 30mL, pregelatinize, add 11.7U CGTase, react at 50°C for 24h, and inactivate with boiling water for 30min. Using the above reaction solution as a substrate, add 5.3U cyclodextrin hydrolase, and react at 65°C for 12h. After the reaction, inactivate with 15mL of 1M NaOH, and neutralize the solution with 1M hydrochloric acid.

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Abstract

The invention discloses a method for purifying gamma-cyclodextrin by utilizing cyclodextrin hydrolase and belongs to the technical field of processing of cyclodextrin. The method comprises the following steps of using starch or dextrin as a substrate, using buffer solution to prepare substrate solution with certain concentration, adding a certain amount of cyclodextrin glucosyltransferase with gamma-CD as a product, controlling reaction temperature and reaction time, and after reaction is ended, killing enzyme; using the reaction liquid as a substrate, adding a certain amount of cyclodextrin hydrolase, controlling reaction temperature and reaction time, and after reaction is ended, killing the enzyme. The method disclosed by the invention utilizes the cyclodextrin hydrolase with selectivedegradability to the cyclodextrin and utilizes a higher degradability of the cyclodextrin hydrolase to alpha-CD and beta-CD and a weaker degradability to gamma-CD to selectively remove the alpha-CD and the beta-CD and retain the gamma-CD so as to improve the purity of the gamma-CD. The method disclosed by the invention breaks through the original methods mostly adopting an organic solvent for precipitation in the industry, adopts the method for purifying the gamma-cyclodextrin by an enzymatic method, is simple in operation, is environment-friendly and has good application potential in the field of cyclodextrin processing.

Description

technical field [0001] The present invention relates to a method for purifying γ-cyclodextrin by using cyclodextrin hydrolase, in particular to a method for γ-cyclodextrin (γ-CD ) purification method, which belongs to the technical field of cyclodextrin processing. Background technique [0002] Cyclodextrin (CD for short) is a general term for a series of cyclic oligosaccharides produced by starch under the action of cyclodextrin glucosyltransferase (CGTase), usually containing 6-12 D-glucopyranose units. Among them, molecules with 6, 7, and 8 glucose units linked by α-1,4 glycosidic bonds, which are studied more and have important practical significance, are called α-cyclodextrin (α-CD), β-cyclodextrin, respectively. Dextrin (beta-CD) and gamma-cyclodextrin (gamma-CD). [0003] Natural β-CD is the most widely used cyclodextrin in industrial production. At present, β-CD accounts for about 70% of the global cyclodextrin production, α-CD accounts for about 15%, and γ-CD onl...

Claims

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Application Information

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IPC IPC(8): C08B37/16
CPCC08B37/0012
Inventor 金征宇柏玉香纪杭燕
Owner JIANGNAN UNIV
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