Preparation method for improving yield of beta-cyclodextrin

A cyclodextrin and glucose-based technology, which is applied in the field of preparation to increase the yield of β-cyclodextrin, can solve the problems of increasing the yield of cyclodextrin, reducing product inhibition, and affecting the final yield, so as to improve the yield of enzymes and bacteria. Utilization rate, load avoidance, and yield improvement effects

Pending Publication Date: 2020-12-08
JIANGSU OGO BIOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the addition of organic solvents can reduce product inhibition and increase the yield of cyclodextrins, the industrial application of cyclodextrins is limited to a certain extent due to its toxicity and danger.
Moreover, cyclodextrin glucosyltransferase has the functions of catalyzing cyclization, coupling and hydrolysis at the same time. It means that cyclodextrin will be hydrolyzed when the cyclization is not dominant, and the presence of small molecular sugars inhibits the cyclization reaction and promotes the coupling and hydrolysis reactions, thereby affecting the final yield
In addition, the enzymatic process has the disadvantages of one-time use of free enzymes, which cannot be recycled and reused, and the free enzymes are mixed with the product, which increases the burden of separation and purification.

Method used

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  • Preparation method for improving yield of beta-cyclodextrin
  • Preparation method for improving yield of beta-cyclodextrin
  • Preparation method for improving yield of beta-cyclodextrin

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preparation example Construction

[0020] The invention provides a preparation method for improving the yield of β-cyclodextrin, comprising the following steps:

[0021] 1) Preparation of immobilized cyclodextrin glucosyltransferase and immobilized cyclodextrin glucosyltransferase producing bacteria:

[0022] Dissolve Eudragit S-100 in ethanol solution, the concentration of Eudragit S-100 is 15-30g / L, add cyclodextrin glucosyltransferase containing 50-70U / mL and 5-10cfu / mL of cyclodextrin glucosyltransferase-producing bacteria, mix evenly, and incubate for 1-3 hours at 25-35°C and pH 6.0-8.0; after incubation, adjust the pH of the solution to <6.0 with a pH regulator, The suspension was obtained, centrifuged at 6000r / min for 30 minutes, and filtered to obtain solid microspheres: add ethanol solution to wash the microspheres, and the suspension was centrifuged at 6000r / min for 30 minutes, and the washing process was repeated three times to obtain immobilized cyclodextrin glucosyltransferase and immobilized cyc...

Embodiment 1

[0027] This implementation provides a preparation method for improving the yield of β-cyclodextrin, comprising the following steps:

[0028] 1) Preparation of immobilized cyclodextrin glucosyltransferase and immobilized cyclodextrin glucosyltransferase producing bacteria:

[0029] After dissolving Eudragit S-100 with ethanol solution, the concentration of Eudragit S-100 is 15g / L, and the cyclodextrin glucosyltransferase containing 50U / mL and 10cfu / mL cyclodextrin glucose are added in a volume ratio of 1:4 base transferase-producing bacteria, mix well, and incubate for 1-3 hours at 25-35°C and pH 6.0-8.0; after incubation, adjust the pH of the solution to <6.0 with ammonium sulfate solution, and let stand to obtain a suspension. Centrifuge at 6000r / min for 30 minutes and filter to obtain solid microspheres: add ethanol solution to wash the microspheres, and centrifuge the suspension at 6000r / min for 30 minutes. The washing process is repeated three times to obtain immobilized c...

Embodiment 2

[0033] This embodiment provides a preparation method for increasing the yield of β-cyclodextrin, comprising the following steps:

[0034] 1) Preparation of immobilized cyclodextrin glucosyltransferase and immobilized cyclodextrin glucosyltransferase producing bacteria:

[0035] Dissolve Eudragit S-100 with ethanol solution, the concentration of Eudragit S-100 is 20g / L, add cyclodextrin glucosyltransferase containing 58U / mL and 9cfu / mL cyclodextrin glucosyl Transferase-producing bacteria, mix evenly, and incubate for 1-3 hours at 25-35°C, pH 6.0-8.0; after incubation, adjust the pH of the solution to <6.0 with ammonium nitrate, and let stand to obtain a suspension, 6000r / Centrifuge for 30 minutes, filter to obtain solid microspheres: add ethanol solution to wash the microspheres, and centrifuge the suspension at 6000r / min for 30 minutes, repeat the washing process three times, and obtain immobilized cyclodextrin glucosyltransferase and immobilized cyclodextrin glucose The bas...

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Abstract

The invention provides a preparation method for improving the yield of beta-cyclodextrin, and belongs to the technical field of cyclodextrin production. Immobilized cyclodextrin glucosyltransferase and immobilized cyclodextrin glucosyltransferase producing bacteria are prepared by using a reversible soluble pH sensitive polymer carrier Eudragit S-100, and the prepared immobilized cyclodextrin glucosyltransferase and immobilized cyclodextrin glucosyltransferase producing bacteria are used for producing beta-cyclodextrin, so that the separation of enzyme and bacteria from products is realized, and the stability and reusability of the enzyme and bacteria are improved; in addition, the cyclodextrin glucosyltransferase producing bacteria are added into a reaction system, glucose generated by enzyme reaction can be utilized, ethanol is generated, the coupling reaction and hydrolysis reaction of the cyclodextrin glucosyltransferase are inhibited, and the cyclization reaction of the cyclodextrin glucosyltransferase is promoted; and meanwhile, the cyclization reaction of the cyclodextrin glucosyltransferase and the growth of the cyclodextrin glucosyltransferase producing bacteria to generate the cyclodextrin glucosyltransferase are synergistic, so that the yield of p-cyclodextrin is improved.

Description

technical field [0001] The invention relates to the technical field of cyclodextrin production, in particular to a preparation method for improving the yield of β-cyclodextrin. Background technique [0002] Cyclodextrin is produced by D-glucopyranose. - Cyclic compounds linked by 1,4-glucoside bonds, among which α-, β- and γ-cyclodextrins composed of 6, 7 and 8 glucose units are the most common. Cyclodextrin can form clathrates with many hydrophobic molecules, thereby changing their physical and chemical properties. Therefore, it is widely used in food, medicine, chemical industry, agriculture and other fields. α, β and γ cyclodextrins are all white crystal powders, among which β-cyclodextrin has the lowest water solubility, so it is easy to crystallize and purify, so β-cyclodextrin is currently produced and widely used in industry. [0003] The industrialized production of cyclodextrin mainly adopts enzymatic process, that is, under the catalysis of cyclodextrin glucosylt...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/087C12P19/18C12P19/04C12R1/865
CPCC12N11/087C12N9/1074C12P19/18C12P19/04C12Y204/01019
Inventor 楼志华张博张培东
Owner JIANGSU OGO BIOTECH
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