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Preparation method of low-molecular chondroitin sulfate for treating myocarditis

A chondroitin sulfate and low-molecular technology, which is applied in the field of preparation of low-molecular chondroitin sulfate, can solve problems such as increasing production cost, and achieve the effects of low production cost, convenient operation and high purity

Inactive Publication Date: 2014-02-26
QINGDAO BETTER BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] However, the enzymatic hydrolysis method has high requirements on the enzymes used, and requires specific chondroitinase. However, no chondroitinase-producing bacteria with independent intellectual property rights have been developed in China, and they can only be imported from abroad at high prices, thereby greatly increasing cost of production

Method used

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  • Preparation method of low-molecular chondroitin sulfate for treating myocarditis

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] The fermentation condition of Arthrobacter sulfonephagous is , and the optimum seed medium components for fermentation are (w / v): chondroitin sulfate 0.8%, KH 2 PO 4 0.02%, yeast extract powder 0.6%, MgSO 4 ·7H 2 0 content 0.8%, pH=6.

[0026] The optimal fermentation medium condition is: put 1.5 liters of seed bacteria solution into a 100 liter fermenter at 25° C., 200 r / min shaking and aeration for 72 hours.

[0027] The optimal conditions for enzymatic hydrolysis of chondroitin sulfate by fermentation broth are to hydrolyze 30 kg of chondroitin sulfate with 100 liters of fermentation broth in an enzymolysis tank, enzymolysis time is 8 hours, enzymolysis temperature is 35°C, and enzymolysis pH=7.

[0028] After the enzymatic hydrolysis, the low molecular weight chondroitin sulfate was prepared by spray drying. The content of chondroitin sulfate disaccharide was 99%, its molecular weight was 458, the content of chondroitin sulfate A was 85%, and the content of chon...

Embodiment 2

[0030] The fermentation conditions of Arthrobacter sulfonephagous are as follows. The optimal seed medium components for 10 liters of fermentation are (w / v): chondroitin sulfate 80g, KH 2 PO 4 2g, yeast extract powder 60g, MgSO 4 ·7H 2 0 content 80g, pH=6.

[0031] The optimal fermentation medium condition is: put 10 liters of seed bacteria solution into a 1000 liter fermenter at 30°C, 250r / min shaking and aeration culture for 80h.

[0032] The optimal conditions for enzymatic hydrolysis of chondroitin sulfate by fermentation broth are to hydrolyze 400 kg of chondroitin sulfate with 1000 liters of fermentation broth in an enzymolysis tank, enzymolysis time is 7 hours, enzymolysis temperature is 40°C, and enzymolysis pH=7.

[0033] Spray drying after enzymolysis finishes and prepares 400 kilograms of low-molecular-weight chondroitin sulfates, the content of chondroitin sulfate disaccharides is 98.5%, its molecular weight is 461, the content of chondroitin sulfate A is 83%, ...

Embodiment 3

[0035] The effect of chondroitin disaccharide on normal H9c2 cardiomyocytes was determined by microscopic observation and MTT method. The results showed that chondroitin disaccharide had no cytotoxicity, and promoted the growth and proliferation of H9c2 cells to a certain extent, and its optimal concentration was 200μgmL -1 .

[0036] The protective effect of different molecular weight CS on the damaged myocardium was evaluated. The results showed that low molecular weight chondroitin sulfate can significantly promote the growth and recovery of damaged myocardial tissue. Chondroitin sulfate and its middle molecule chondroitin sulfate can promote the recovery of the heart, but the effect is not as obvious as that of low molecular weight chondroitin sulfate.

[0037] After gavage for 7 days, the injury of cardiomyocytes in the CS group was slightly alleviated, and the arrangement of myocardial fibers, the degree of muscle fiber atrophy, the width of the septum, the cytoplasm a...

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Abstract

The invention discloses a preparation method of low-molecular chondroitin sulfate for treating myocarditis. The preparation method comprises the step of fermenting raw materials through arthrobacter sulfonivorans fermentation liquor, carrying out enzymolysis treatment on the chondroitin sulfate in the raw materials to obtain the low-molecular chondroitin sulfate, wherein the content of the chondroitin sulfate disaccharide in the prepared low-molecular chondroitin sulfate is more than 97%, the molecular weight of the chondroitin sulfate disaccharide is between 450-480, the chondroitin sulfate A content is between 70-95%, and the chondroitin sulfate C content is between 5-25%. The preparation method is convenient to operate, short in period and low in cost, the prepared low-molecular chondroitin sulfate is good in curative effect and high in purity, and can be applied to large-scale industrial production; the low-molecular chondroitin sulfate has obvious effect on myocardial cell injury repair through rat test.

Description

technical field [0001] The invention relates to a preparation method of chondroitin sulfate, in particular to a preparation method of low molecular weight chondroitin sulfate for treating myocarditis. Background technique [0002] Chondroitin Sulfate (CS) is an acidic mucopolysaccharide present in connective tissues of humans and animals, mainly distributed in cartilage, bone, tendon, sarcolemma and blood vessel walls, composed of D-glucuronic acid and N-acetyl -D-Galactosamine is connected by 1,3 glycosidic bonds to form disaccharides, and the disaccharides are connected by β-1,4 glycosidic bonds. The relative molecular mass (Mr) of CS is generally above 20×103, and when Mr decreases to less than 10×103, it is often called low molecular weight chondroitin sulfate (LMWCS). [0003] As a component of proteoglycans, CS exists in intracellular granules, cell membranes, basement membranes and extracellular matrices. It binds specifically to cytokines, adhesion factors or their ...

Claims

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Application Information

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IPC IPC(8): C12P19/26C12R1/06
Inventor 曹荣军杨庆利刘万顺王凤山张天民余玉奎台文静陈杰刘少芳
Owner QINGDAO BETTER BIO TECH
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