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Method for preparing injection-grade alpha-cyclodextrin

A cyclodextrin, injection-grade technology, applied in the field of biochemistry, can solve the problems of organic solvent residue, low product quality, strong reaction, etc., and achieve the effect of low production cost, high reactivity, and promotion of adsorption

Inactive Publication Date: 2014-08-13
JINAN KANGHE MEDICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The invention provides a preparation method of injection-grade α-cyclodextrin, which uses epoxy resin as an immobilized carrier to obtain stable α-CGTase immobilized enzyme, and uses a solvent-free method to prepare α-cyclodextrin, which can Effectively overcome the problems of high energy consumption, strong reaction, residual organic solvent and low product quality in the current technology

Method used

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  • Method for preparing injection-grade alpha-cyclodextrin
  • Method for preparing injection-grade alpha-cyclodextrin
  • Method for preparing injection-grade alpha-cyclodextrin

Examples

Experimental program
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Effect test

Embodiment 1

[0038] step 1:

[0039] Genetically engineered bacteria E.coli BL21(DE3) / pet24a(+) (China Center for Type Microorganisms Collection, CCTCCM208063) inoculated in LB medium, cultured at 30-37°C for 6-12 hours, then transferred to TB medium, cultured at 30-37°C for 2-6 hours, added 0.01% IPTG and cooled Cultivate at 20-28°C for 30-36 hours to obtain a fermentation broth. The fermented liquid was taken and centrifuged at 6000rpm for 20min in a high-speed refrigerated centrifuge to obtain a supernatant. The supernatant was taken to measure the enzyme activity according to the method of A Spectrophotometric Assay for the Cyclization Activity of Cyclomaltohexaose (α-Cyclodextrin) Glucanotransferase in the literature, and the cyclization activity was 50 U / ml.

[0040] Take the supernatant, adjust the pre-membrane pressure of the ultrafiltration membrane unit to 1MPa, and the post-membrane pressure to 0.4MPa, and then enter the ultrafiltration membrane filtration system (the cut-o...

Embodiment 2

[0057] step 1:

[0058] Bacillus softening ( Bacillus macerans ) inoculated in LB medium, cultured at 30-37°C for 6-12 hours, then transferred to fermentation medium (g / L): corn steep liquor 60, corn flour 20, dipotassium hydrogen phosphate 1.5, magnesium sulfate 0.2, sodium carbonate 10, pH 7.0, cultured at 35°C for 30-36 hours to obtain a fermentation broth. The fermented liquid was taken and centrifuged at 6000rpm for 20min in a high-speed refrigerated centrifuge to obtain a supernatant. The supernatant was taken to measure the enzyme activity according to the method in Example 1, and the cyclization activity was 30 U / ml.

[0059] Take the supernatant, adjust the pre-membrane pressure of the ultrafiltration membrane unit to 1MPa, and the post-membrane pressure to 0.4MPa, then enter the ultrafiltration membrane filtration system (the cut-off pore size of the ultrafiltration membrane is 10000), the working temperature is 30 ° C ~ 40 ° C, water and Some small molecular su...

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Abstract

The invention discloses a method for preparing injection-grade alpha-cyclodextrin. The method comprises the steps: preparing alpha-CGTase immobilized enzyme by adopting epoxy resin as a vector, preparing starch into starch milk, adding the alpha-CGTase immobilized enzyme for liquefying starch, reacting in a filled reactor filled with the alpha-CGTase immobilized enzyme, carrying out ultrafiltration on a conversion solution, and collecting dialysate; treating the dialysate by using alpha-amylase and saccharifying enzyme, decolorizing with activated carbon, concentrating by using reverse osmosis membrane, recrystallizing, filtrating to remove beta-cyclodextrin; and further concentrating mother liquor with a reverse osmosis membrane, crystallizing, filtrating, drying and grinding, thus obtaining the injection-grade alpha-cyclodextrin. According to the method, the prepared immobilized enzyme has the advantages that the stability is good, the immobilized enzyme can be reused; a product and a substrate can be seperated extremely easy, the utilization efficiency of enzymes is high, continuous operation and automated control can be carried out, the process is simple and convenient, the production cost is low; alpha-cyclodextrin is produced by adopting a solvent-free process, the production equipment is simple, the reaction conditions are mild, the energy consumption is low, and solvent residual contamination does not exist, so that the method is very environment-friendly and energy-saving.

Description

technical field [0001] The invention belongs to the technical field of biochemistry, and in particular relates to a preparation method of injection-grade alpha-cyclodextrin. technical background [0002] Cyclodextrin is a kind of cyclic oligomeric compound formed by the enzymatic hydrolysis of starch or starch matrix by cyclodextrin glucosyltransferase, which is formed by connecting D-glucopyranose units through α-1,4-glucosidic bonds. According to the number of glucose units, cyclodextrins can be divided into α-, β-, γ-, δ-cyclodextrins, etc., among which the most common ones are α-, β-, γ with a degree of polymerization of 6, 7, 8 - Cyclodextrins. [0003] The three-dimensional structure of cyclodextrin is a hollow cylinder, and there are non-polar C3 and C5 hydrogens and oxygen atoms connected by ether bonds inside the cylinder, so the inner cavity of the cylinder is hydrophobic; while the glucose The secondary hydroxyl groups on C2 and C3 are located at the wide side o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/14C12P19/04
CPCY02P20/10
Inventor 张颖刘鹏姜寿俊孙健高国锐刘元鑫孙剑峰王学斌葛庆燕闫飞刘骏郭凤霞徐丹
Owner JINAN KANGHE MEDICAL TECH
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