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Method and application for refining glucosamine hydrochloride from micro-organism fermentation liquid

A technology of glucosamine hydrochloride and microbial fermentation liquid, which is applied in the field of refined glucosamine hydrochloride, can solve the problems of low glucosamine extraction rate and long cycle, achieve high product purity, improve recovery rate, and facilitate efficient separation and purification Effect

Inactive Publication Date: 2019-07-19
ZHEJIANG CHINESE MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] In view of this, the technical problem to be solved by the present invention is to provide a separation and purification process of glucosamine hydrochloride with reasonable process, high recovery rate and high product purity for the deficiencies in the prior art, which solves the problem of glucosamine hydrochloride in the prior art. The problem of low glucosamine extraction rate and long cycle; and provides a new application of refined glucosamine hydrochloride against pancreatic cancer in vivo

Method used

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  • Method and application for refining glucosamine hydrochloride from micro-organism fermentation liquid
  • Method and application for refining glucosamine hydrochloride from micro-organism fermentation liquid
  • Method and application for refining glucosamine hydrochloride from micro-organism fermentation liquid

Examples

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Embodiment 1

[0043] 1. Obtaining of glucosamine microbial fermentation liquid

[0044] The glucosamine fermentation broth used in this example is obtained by fermentation of the prior art method, that is, the genetic engineering strain BL21(DE3)-pET28a-ES-glmS-GAN1BL21 of Escherichia coli is inoculated into the fermentation medium for fermentation, and the obtained product contains Fermentation broth of glucosamine and N-acetylglucosamine.

[0045] For details, please refer to the patent application text with publication number CN108588049A titled "A Glucosamine Synthetase, Engineering Bacteria and Its Application"; wherein, the preparation process of engineering bacteria refers to Examples 1 and 2, and the process of fermenting and preparing glucosamine refers to the implementation Example 5.

[0046] 2. Refining of glucosamine hydrochloride

[0047] (1) Get the glucosamine microbial fermentation liquid, after centrifuging at 12000rpm for 10min, remove the bacteria, collect the supernatan...

Embodiment 2

[0071] Example 2 In vivo anti-pancreatic cancer activity of glucosamine hydrochloride

[0072] Using the glucosamine hydrochloride prepared in Example 1 as the raw material drug, its effect on pancreatic cancer was systematically evaluated in vivo.

[0073] The mouse-derived Pan02 pancreatic cancer cells were used to establish a subcutaneous xenograft model of pancreatic cancer in C57BL / 6 black mice. Pan02 cells were resuspended in RPMI 1640 medium containing 10% fetal bovine serum, and incubated at 37°C and 5% CO 2 Cultivate in a cell incubator. After the cells adhere to the wall and grow nearly 80%, discard the culture medium in the culture bottle, wash it with PBS and add an appropriate amount of trypsin to digest it. When the adherent cells become round, add the culture medium to stop and wait for 800r. Centrifuge for 5 min at 1 / min, and take an appropriate proportion of single cells for subculture. Amplify Pan02 cells. When the cell confluence of each culture flask reach...

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Abstract

The present invention discloses a method and an application for refining glucosamine hydrochloride from micro-organism fermentation liquid. The refining method comprises the following steps: diluted hydrochloric acid is added to micro-organism fermentation liquid after bacteria are removed, acid hydrolysis is conducted in a boiling water bath, then activated carbon is added for adsorption, then alkali is added to adjust pH to neutral, a polypropylene filter membrane is used for ultrafiltration treatment, and finally treatment liquid is successively subjected to vacuum concentration, low-temperature crystallization, washing and vacuum drying to obtain refined amino acid hydrochloride. The "acid hydrolysis, activated carbon adsorption, ultrafiltration, concentration, crystallization, washingand drying refining technology is used to separate and purify the specific micro-organism fermentation liquid containing N-acetyl D-glucosamine and D-glucosamine to obtain the glucosamine hydrochloride with high recovery rate and high product purity, and solves problems of relatively low glucosamine extraction rate and long extraction period in the micro-organism fermentation liquid in the priorart.

Description

technical field [0001] The invention relates to the technical field of separation and extraction of microbial fermentation products, in particular to a method and application for refining glucosamine hydrochloride from microbial fermentation liquid. Background technique [0002] Glucosamine hydrochloride, also known as D-glucosamine hydrochloride, is a natural mucopolysaccharide, an odorless but slightly bitter white crystalline powder, easily soluble in water but insoluble in ethanol. Glucosamine hydrochloride is widely used in the fields of chemical industry, medicine, food and health care. Glucosamine can specifically act on articular cartilage, restore the normal metabolic function of chondrocytes, stimulate chondrocytes to produce proteoglycans with normal polymer structure, inhibit enzymes that damage cartilage, and delay the pathological process and disease of osteoarthritis It can improve joint activity and relieve pain, so it is widely used in the treatment of oste...

Claims

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Application Information

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IPC IPC(8): C07H5/06C07H1/08A61P35/00
CPCA61P35/00C07H1/08C07H5/06
Inventor 程汝滨葛宇清方昀
Owner ZHEJIANG CHINESE MEDICAL UNIVERSITY
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