Low-molecular-weight chondroitin sulfate and preparation method thereof

A technology of chondroitin sulfate and low molecular weight, which is applied in the preparation of sugar derivatives, chemical instruments and methods, and medical preparations containing active ingredients, etc. It can solve the problems of insufficient research, narrow range of molecular weight distribution, and low catalytic efficiency. , to achieve the effects of short production cycle, narrow molecular weight distribution range and high enzyme activity

Inactive Publication Date: 2020-10-02
NANJING HANXIN PHARMA TECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The journal document "Enzymatic Preparation of Chondroitin Sulfate Oligosaccharides and Its Antioxidant Activity" Food Industry Science and Technology, 2017, 13, 48-52, discloses the use of chondroitin sulfate enzyme (molecular weight 76kDa) after fermentation from the enzyme-producing strain Acinetobacter sp.C26 ), the catalytic efficiency is low, the reported reaction concentration is only 2%, and the ratio of oligosaccharides has not been analyzed; the cracking has obtained chondroitin sulfate disaccharides and tetrasaccharides, and m/Z is 342 and 458 are disaccharides, m/ What Z is 939 is a tetrasaccharide, which is different from our low molecular weight chondroitin sulfate in composition and molecular weight. For example, in our process, the molecular weight of disaccharide is about 379, 459 and the molecular

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  • Low-molecular-weight chondroitin sulfate and preparation method thereof
  • Low-molecular-weight chondroitin sulfate and preparation method thereof
  • Low-molecular-weight chondroitin sulfate and preparation method thereof

Examples

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Example Embodiment

[0054] Example 1 Enzymatic hydrolysis reaction

[0055] In a 5L glass beaker, add 2L of purified water, control the stirring speed to 400rpm, add 800g of shark bone chondroitin sulfate, adjust the pH to 7.0 with sodium hydroxide solution, and add 200U / L of chondroitin sulfate lyase. Keep the system at 30°C and stir the reaction. After 6 hours of reaction, check whether the average molecular weight is lower than 1000 Da. If the reaction is not complete, continue to control after 4 hours of reaction time. Continue the reaction until the average molecular weight is less than 1000 Da, and it is judged as qualified.

Example Embodiment

[0056] Example 2 Enzymatic hydrolysis reaction

[0057] In a 5L glass beaker, add 2L of purified water, control the stirring speed to 700rpm, add 400g of shark bone chondroitin sulfate, adjust the pH to 6.5 with sodium hydroxide solution, and add 300U / L of chondroitin sulfate lyase. Keep the system at 35°C and stir to react. After 6 hours of reaction, check whether the average molecular weight is lower than 1000 Da. If the reaction is not complete, continue to control after 4 hours of reaction time. Continue the reaction until the average molecular weight of the hydrolyzate is less than 1000 Da, and it is judged as qualified.

Example Embodiment

[0058] Example 3 Enzymatic hydrolysis reaction

[0059] In a 5L glass beaker, add 2L of purified water, control the stirring speed to 100rpm, add 200g of shark bone chondroitin sulfate, after all is dissolved, adjust the pH to 8.0 with sodium hydroxide solution, add 100U / L of chondroitin sulfate lyase, Keep the system at 25°C and stir the reaction. After 6 hours of reaction, check whether the average molecular weight is lower than 1000 Da. If the reaction is not complete, continue the central control after prolonging the reaction time for 4 hours. Continue the reaction until the average molecular weight of the hydrolyzate is less than 1000 Da, and it is judged as qualified.

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Abstract

The invention discloses a low-molecular-weight chondroitin sulfate and a preparation method thereof. Macromolecular chondroitin sulfate is used as a raw material, and the low-molecular-weight chondroitin sulfate having average molecular weight of less than 1000 daltons is produced through a production process comprising chondroitin sulfate lyase degradation, deproteinization, filtration sterilization, drying, and the like. According to the low-molecular-weight chondroitin sulfate, the molecular weight distribution range is narrow, the content of chondroitin sulfate disaccharide is 43-60%, thecontent of chondroitin sulfate tetrasaccharide is 30-45%, the sum of contents of the chondroitin sulfate disaccharide and the chondroitin sulfate tetrasaccharide is greater than 87%, the total contentof oligosaccharides in the low-molecular-weight chondroitin sulfate is 97% or above, and the protein content is not more than 0.5%. Compared with common marketed macromolecular chondroitin sulfate, the product has a more obvious repairing effect on chondrocytes damaged by 1mM hydrogen peroxide at a concentration of 50-100 [mu]g/mL, has a strong repairing ability and a repair rate between 14% and23%, can be used to treat joint injuries, and is an important raw material for medical products, healthcare products, cosmetics, food, etc..

Description

technical field [0001] The invention belongs to the technical field of biochemical industry, and in particular relates to a low molecular weight chondroitin sulfate and a preparation method thereof. Background technique [0002] Chondroitin sulfate (Chondroitin sulfate, CS, macromolecular chondroitin sulfate or chondroitin sulfate polysaccharide) is a kind of linear polysaccharide containing polyanions, composed of D-glucuronic acid (GlcA) and N-acetyl-D-amino semi Lactose (GalNAc) is connected by β-1,3 glycosidic bonds to form disaccharide units, and the disaccharide units are connected by β-1,4 glycosidic bonds, and sulfate groups are introduced at different positions during the subsequent biosynthesis process. Widely present in the cartilage and connective tissue of various animals. [0003] A large number of studies have shown that CS has the activities of lowering blood fat, anti-atherosclerosis, enhancing immunity, anti-viral hepatitis and anti-tumor. field. For exa...

Claims

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Application Information

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IPC IPC(8): C07H13/04C07H5/06C07H1/00C08B37/08A61P19/02C12P19/26A23L33/125A61K8/73A61K8/60A61Q19/00
CPCA23V2002/00A61K8/60A61K8/73A61K2800/10A61Q19/00A23L33/125A61P19/02C07H1/00C07H5/06C07H13/04C08B37/0069C12P19/26A23V2200/30A23V2250/51
Inventor 王印崔怀言梁旭光李德耿徐勇刚汤传根陈松张昊宁
Owner NANJING HANXIN PHARMA TECH CO LTD
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