Method for producing low-molecular chondroitin sulfate by catalytic process

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 chondroitin sulfate, is a kind of polymeric acidic mucopolysaccharide made from the cartilage of animals (pigs, cattle, horses, sheep, chickens, sharks, etc.), and its molecular structure is β-D-glucuronic acid It is a polysaccharide chain composed of repeating disaccharide units formed by β1→3 glycosidic bonds with N-acetylgalactosamine. 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 relatively 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 m...

Examples

Embodiment 1

[0023] Example 1: Production of low molecular chondroitin sulfate according to the following steps

[0024] (1) Boiled: 100 kilograms of cartilage materials are immersed in water, heated to 80°C, and boiled for 24 hours to peel off the muscle tissue attached to the cartilage surface and remove the cartilage;

[0025] (2) Steaming: 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) Enzymatic hydrolysis: 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, add 1 kg of alkaline protease, and keep the pH at 8. Keep at 55°C and stir 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 solution alcohol concentration reach 60%, and settle for more than 1...

Embodiment 2

[0032] Example 2: Production of low molecular chondroitin sulfate according to the following steps

[0033] (1) Boiled: 100 kg of cartilage raw materials are immersed in water, heated to 90°C, and boiled for 18 hours to peel off the muscle tissue attached to the cartilage surface and remove the cartilage;

[0034] (2) Steaming: 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) Enzymatic hydrolysis: 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 maintain pH 8. .5, keep 57.5℃ and stir for 5 hours, heat to boiling for 15 minutes, and filter;

[0036] (4) Alcohol precipitation: adjust the pH of the filtrate obtained in step (3) to 6.5, and then add alcohol to make the solution alcohol concentration reach 60%, and preci...

Embodiment 3

[0041] Example 3: Production of low molecular chondroitin sulfate according to the following steps

[0042] (1) Boiled: 100 kilograms of cartilage raw materials are immersed in water, heated to 100°C, and boiled for 12 hours to peel off the muscle tissue attached to the cartilage surface and remove the cartilage;

[0043] (2) Steaming: 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) Enzymatic hydrolysis: 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 9 , Keep at 60℃ 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, and then add alcohol to make the solution alcohol concentration reach 60%, and prec...

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 chondroitin sulfate, is a kind of polymeric acidic mucopolysaccharide made from the cartilage of animals (pigs, cattle, horses, sheep, chickens, sharks, etc.), and its molecular structure is β-D-glucuronic acid It is a polysaccharide chain composed of repeating disaccharide units formed by β1→3 glycosidic bonds with N-acetylgalactosamine. 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 relatively 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 m...

Examples

Embodiment 1

[0023] Example 1: Production of low molecular chondroitin sulfate according to the following steps

[0024] (1) Boiled: 100 kilograms of cartilage materials are immersed in water, heated to 80°C, and boiled for 24 hours to peel off the muscle tissue attached to the cartilage surface and remove the cartilage;

[0025] (2) Steaming: 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) Enzymatic hydrolysis: 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, add 1 kg of alkaline protease, and keep the pH at 8. Keep at 55°C and stir 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 solution alcohol concentration reach 60%, and settle for more than 1...

Embodiment 2

[0032] Example 2: Production of low molecular chondroitin sulfate according to the following steps

[0033] (1) Boiled: 100 kg of cartilage raw materials are immersed in water, heated to 90°C, and boiled for 18 hours to peel off the muscle tissue attached to the cartilage surface and remove the cartilage;

[0034] (2) Steaming: 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) Enzymatic hydrolysis: 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 maintain pH 8. .5, keep 57.5℃ and stir for 5 hours, heat to boiling for 15 minutes, and filter;

[0036] (4) Alcohol precipitation: adjust the pH of the filtrate obtained in step (3) to 6.5, and then add alcohol to make the solution alcohol concentration reach 60%, and preci...

Embodiment 3

[0041] Example 3: Production of low molecular chondroitin sulfate according to the following steps

[0042] (1) Boiled: 100 kilograms of cartilage raw materials are immersed in water, heated to 100°C, and boiled for 12 hours to peel off the muscle tissue attached to the cartilage surface and remove the cartilage;

[0043] (2) Steaming: 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) Enzymatic hydrolysis: 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 9 , Keep at 60℃ 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, and then add alcohol to make the solution alcohol concentration reach 60%, and prec...