Method for producing low molecular weight chondroitin sulfate by catalytic method

Abstract

The invention discloses a method for producing low-molecular chondroitin sulfate by a catalytic process, which comprises the following steps: (1) boiling with water; (2) steaming; (3) enzymolysis; (4) alcohol precipitation; (5) catalysis; (6) ultrafiltration; (7) alcohol precipitation; and (8) dehydration and drying. By using the catalyst Cu / ZSM-5 molecular sieve to degrade the chondroitin sulfate, the method has the advantages of mild and controllable conditions, fewer residual metal ion impurities in the degraded solution and no need of ion exchange resin removal, is simple to operate in the production process, and is suitable for large-scale production; and the product quality is stable, and the chondroitin sulfate recovery rate is up to higher than 80%.

Description

technical field

[0001] The invention belongs to the field of biotechnology, in particular to a method for producing low-molecular chondroitin sulfate by catalytic method. Background technique

[0002] Chondroitin sulfate, English name chondroitinsulfate, is a kind of polymeric acidic mucopolysaccharide based on the cartilage of animals (pigs, cattle, horses, sheep, chickens, sharks, etc.), and its molecular structure is β-D-glucuronic acid and N-acetylgalactosamine is a polysaccharide chain composed of repeating disaccharide units formed by β1→3 glycosidic bonds. It is a white or slightly yellow powder with strong water absorption, easily soluble in water to form a viscous liquid, insoluble in organic solvents such as alcohol, acetone, ether and chloroform. Its salts are more stable to heat. The biological activity of chondroitin sulfate varies with molecular weight. Its pharmacological activity is closely related to its molecular weight, and the size of its molecular wei...

Examples

Embodiment 1

[0023] Embodiment 1: produce low molecular weight chondroitin sulfate according to the following steps

[0024] (1) Poaching: Submerge 100 kg of cartilage raw materials in water, raise the temperature to 80°C, and cook for 24 hours to peel off the muscle tissue attached to the surface of the cartilage, and remove the cartilage;

[0025] (2) Cooking: Put the boiled cartilage obtained in step (1) into a kettle, steam at 100°C for more than 4 hours, and collect the upper layer of oil;

[0026] (3) Enzymolysis: Submerge the softened cartilage obtained in step (2) with 100 liters of water, add 3 kg of sodium chloride, adjust the pH to 8 with alkali, raise the temperature to 55°C, and then add 1 kg of alkaline protease to keep the pH at 8. Stir at 55°C for 6 hours, heat up to boiling for 12 minutes, and filter;

[0027] (4) Alcohol precipitation: adjust the pH of the filtrate obtained in step (3) to 6, then add alcohol to make the alcohol concentration of the solution reach 60%, an...

Embodiment 2

[0032] Embodiment 2: produce low molecular weight chondroitin sulfate according to the following steps

[0033] (1) Poaching: Submerge 100 kg of cartilage raw materials in water, raise the temperature to 90°C, and cook for 18 hours to peel off the muscle tissue attached to the surface of the cartilage, and remove the cartilage;

[0034] (2) Cooking: Put the boiled cartilage obtained in step (1) into a kettle, steam at 100°C for more than 4 hours, and collect the upper layer of oil;

[0035] (3) Enzymolysis: Submerge the softened cartilage obtained in step (2) with 100 liters of water, add 3 kg of sodium chloride, adjust the pH to 8.5 with alkali, raise the temperature to 57.5°C, and then add 1.5 kg of 2709 alkaline protease to keep the pH at 8 .5, keep warm at 57.5°C and stir for 5 hours, heat up to boiling for 15 minutes, and filter;

[0036] (4) Alcohol precipitation: Adjust the pH of the filtrate obtained in step (3) to 6.5, then add alcohol to make the alcohol concentrati...

Embodiment 3

[0041] Embodiment 3: produce low molecular weight chondroitin sulfate according to the following steps

[0042] (1) Poaching: Submerge 100 kg of cartilage raw materials in water, raise the temperature to 100°C, and cook for 12 hours to peel off the muscle tissue attached to the surface of the cartilage, and remove the cartilage;

[0043] (2) Cooking: Put the boiled cartilage obtained in step (1) into a kettle, steam at 100°C for more than 4 hours, and collect the upper layer of oil;

[0044] (3) Enzymolysis: Submerge the softened cartilage obtained in step (2) with 100 liters of water, add 3 kg of sodium chloride, adjust the pH to 9 with alkali, raise the temperature to 60°C, and then add 2 kg of 2709 alkaline protease to keep the pH at 9 , keep warm at 60°C and stir for 4 hours, heat up to boiling for 18 minutes, and filter;

[0045] (4) Alcohol precipitation: Adjust the pH of the filtrate obtained in step (3) to 7, then add alcohol to make the alcohol concentration of the s...

technical field

[0001] The invention belongs to the field of biotechnology, in particular to a method for producing low-molecular chondroitin sulfate by catalytic method. Background technique

[0002] Chondroitin sulfate, English name chondroitinsulfate, is a kind of polymeric acidic mucopolysaccharide based on the cartilage of animals (pigs, cattle, horses, sheep, chickens, sharks, etc.), and its molecular structure is β-D-glucuronic acid and N-acetylgalactosamine is a polysaccharide chain composed of repeating disaccharide units formed by β1→3 glycosidic bonds. It is a white or slightly yellow powder with strong water absorption, easily soluble in water to form a viscous liquid, insoluble in organic solvents such as alcohol, acetone, ether and chloroform. Its salts are more stable to heat. The biological activity of chondroitin sulfate varies with molecular weight. Its pharmacological activity is closely related to its molecular weight, and the size of its molecular wei...

Examples

Embodiment 1

[0023] Embodiment 1: produce low molecular weight chondroitin sulfate according to the following steps

[0024] (1) Poaching: Submerge 100 kg of cartilage raw materials in water, raise the temperature to 80°C, and cook for 24 hours to peel off the muscle tissue attached to the surface of the cartilage, and remove the cartilage;

[0025] (2) Cooking: Put the boiled cartilage obtained in step (1) into a kettle, steam at 100°C for more than 4 hours, and collect the upper layer of oil;

[0026] (3) Enzymolysis: Submerge the softened cartilage obtained in step (2) with 100 liters of water, add 3 kg of sodium chloride, adjust the pH to 8 with alkali, raise the temperature to 55°C, and then add 1 kg of alkaline protease to keep the pH at 8. Stir at 55°C for 6 hours, heat up to boiling for 12 minutes, and filter;

[0027] (4) Alcohol precipitation: adjust the pH of the filtrate obtained in step (3) to 6, then add alcohol to make the alcohol concentration of the solution reach 60%, an...

Embodiment 2

[0032] Embodiment 2: produce low molecular weight chondroitin sulfate according to the following steps

[0033] (1) Poaching: Submerge 100 kg of cartilage raw materials in water, raise the temperature to 90°C, and cook for 18 hours to peel off the muscle tissue attached to the surface of the cartilage, and remove the cartilage;

[0034] (2) Cooking: Put the boiled cartilage obtained in step (1) into a kettle, steam at 100°C for more than 4 hours, and collect the upper layer of oil;

[0035] (3) Enzymolysis: Submerge the softened cartilage obtained in step (2) with 100 liters of water, add 3 kg of sodium chloride, adjust the pH to 8.5 with alkali, raise the temperature to 57.5°C, and then add 1.5 kg of 2709 alkaline protease to keep the pH at 8 .5, keep warm at 57.5°C and stir for 5 hours, heat up to boiling for 15 minutes, and filter;

[0036] (4) Alcohol precipitation: Adjust the pH of the filtrate obtained in step (3) to 6.5, then add alcohol to make the alcohol concentrati...

Embodiment 3

[0041] Embodiment 3: produce low molecular weight chondroitin sulfate according to the following steps

[0042] (1) Poaching: Submerge 100 kg of cartilage raw materials in water, raise the temperature to 100°C, and cook for 12 hours to peel off the muscle tissue attached to the surface of the cartilage, and remove the cartilage;

[0043] (2) Cooking: Put the boiled cartilage obtained in step (1) into a kettle, steam at 100°C for more than 4 hours, and collect the upper layer of oil;

[0044] (3) Enzymolysis: Submerge the softened cartilage obtained in step (2) with 100 liters of water, add 3 kg of sodium chloride, adjust the pH to 9 with alkali, raise the temperature to 60°C, and then add 2 kg of 2709 alkaline protease to keep the pH at 9 , keep warm at 60°C and stir for 4 hours, heat up to boiling for 18 minutes, and filter;

[0045] (4) Alcohol precipitation: Adjust the pH of the filtrate obtained in step (3) to 7, then add alcohol to make the alcohol concentration of the s...