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Method for preparing chondroitin sulfate and collagen polypeptide from animal cartilages

A technology of collagen polypeptide and chondroitin sulfate, which is applied to the preparation method of peptides, chemical instruments and methods, peptides, etc., can solve the problems of low content of collagen polypeptides, limited product application fields, and complex components of complexes, etc., and achieve reduction The content of miscellaneous protein, increase the added value of the product, the obvious effect of economic benefit and environmental benefit

Active Publication Date: 2013-11-13
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above-mentioned processes have realized the comprehensive utilization of cartilage to a certain extent, but there are also some shortcomings: (1) the composition of the compound is complex, and the application field of the product is limited; (2) the bone residue after enzymatic hydrolysis is used to extract collagen Polypeptides, the process is cumbersome, and the content of collagen polypeptides in the bone residue is low; (3) Chondroitin sulfate is obtained by alcohol precipitation of the enzymatic hydrolysis solution, and the content of miscellaneous proteins is relatively high
The application of anion exchange technology to the separation and purification of chondroitin sulfate and collagen polypeptide from cartilage hydrolyzate can realize high-value utilization of cartilage. This method has not been reported

Method used

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  • Method for preparing chondroitin sulfate and collagen polypeptide from animal cartilages
  • Method for preparing chondroitin sulfate and collagen polypeptide from animal cartilages
  • Method for preparing chondroitin sulfate and collagen polypeptide from animal cartilages

Examples

Experimental program
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Effect test

Embodiment 1

[0046] The dried shark cartilage (water content 8%) is crushed to an average particle diameter of about 0.1-1mm, and 10kg is accurately weighed, and 0.1kg alkaline protease (200,000 U / g) and 100L water are added respectively, and the mass concentration is Adjust the pH to 9.0 with 40% NaOH solution, and stir at 55°C for 12h. After the enzymatic hydrolysis, the temperature was raised to 70°C and kept for 30 minutes to inactivate the enzyme. After the enzymolysis solution is cooled, filter, get the filtrate, adjust the pH value to 7 with 6mol / L HCl solution, and concentrate by ultrafiltration (the molecular weight cut-off of the ultrafiltration membrane is 3000), to obtain 9.8L of concentrated solution of 20% chondroitin sulfate concentration (potential Titration method to detect the content of chondroitin sulfate in the concentrate). Use 6mol / L HCl solution to adjust the pH of the concentrated solution to 2.0, wet the D202 type anion exchange resin column with a resin volume o...

Embodiment 2

[0052] Grind the dried shark cartilage (water content 8%) to an average particle size of about 0.1-1mm, accurately weigh 10kg, add 0.15kg bromelain (2400GDU / g) and 150L water respectively, adjust with 6mol / L HCl solution pH to 6.0, stirred at 30°C for 4h. After the enzymatic hydrolysis, the temperature was raised to 70°C and kept for 60 minutes to inactivate the enzyme. After the enzymolysis solution is cooled, filter, take the filtrate, adjust the pH value to 7 with a mass concentration of 40% NaOH solution, and concentrate by ultrafiltration (the molecular weight cut-off of the ultrafiltration membrane is 3000), to obtain 95 L of concentrated solution with a concentration of chondroitin sulfate of 2%. Use 6mol / L HCl solution to adjust the pH of the concentrated solution to 3.0, wet the D315 type anion exchange resin column with a resin volume of 57L at a flow rate of 0.5VB / h, and collect the effluent A; then, use 114L pure water at 0.5VB / h The flow rate of h is eluted, the ...

Embodiment 3

[0055] Grind the dried shark cartilage (water content 8%) to an average particle size of about 1-5mm, accurately weigh 10kg, add 0.05kg acid protease (50,000 U / g) and 100L water respectively, and use 6mol / L HCl The pH of the solution was adjusted to 2.0, and stirred at 55 °C for 5 h. After the enzymatic hydrolysis, the temperature was raised to 80°C and kept for 10 minutes to inactivate the enzyme. After cooling the enzymolysis solution, filter it, take the filtrate, adjust the pH value to 7 with a mass concentration of 40% NaOH solution, and concentrate by ultrafiltration (the molecular weight cut-off of the ultrafiltration membrane is 3000), to obtain 38L of a concentrated solution with a concentration of 5% chondroitin sulfate. Use 6mol / L HCl solution to adjust the pH of the concentrated solution to 4.0, wet the D307 type anion exchange resin column with a resin volume of 23L at a flow rate of 1VB / h, and collect the effluent A; then, use 230L pure water at 0.1VB / h The flow...

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Abstract

The invention discloses a method for preparing chondroitin sulfate and collagen polypeptide from animal cartilages, which comprises the following steps: crushing animal cartilage materials to obtain cartilage powder; hydrolyzing the cartilage powder with protease, and then carrying out ultrafiltration and concentration to obtain a concentrated solution; passing the concentrated solution through an anion exchange resin column, and collecting an effluent solution A; eluting with pure water, and collecting a water washing solution B; merging the effluent solution A and the water washing solution B, carrying out ultrafiltration and concentration, and carrying out spray drying to obtain the collagen polypeptide; and eluting the anion exchange resin column with an NaCl solution, carrying out ultrafiltration and concentration on the eluent solution, carrying out alcohol precipitation, dehydrolyzing, and drying to obtain the chondroitin sulfate. The invention realizes the high-valued utilization of cartilage resources, can simultaneously and efficiently extract and separate the chondroitin sulfate and the collagen polypeptide, increases the added values of the products and reduces the environment pollution.

Description

(1) Technical field [0001] The invention relates to a processing technology of animal cartilage, in particular to a process for extracting by enzymatic hydrolysis and using anion exchange resin to directly separate and purify chondroitin sulfate and collagen polypeptide from cartilage enzymatic hydrolysis solution. (2) Background technology [0002] my country is a large agricultural country, and the leftover cartilage resources produced by the processing of various meat products are very rich in many types. Animal cartilage is rich in active ingredients such as chondroitin sulfate and collagen. These bioactive macromolecules are widely used in the fields of medicine, food and cosmetics. Chondroitin Sulfate (CS) is an acidic mucopolysaccharide composed of repeating disaccharide units composed of D-glucuronic acid and N-acetylgalactosamine. It is mainly used clinically to prevent and treat arthritis, neuralgia, and arthralgia. , arteriosclerosis, hearing impairment and hepat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/06C07K1/36C07K1/34C07K1/18C08B37/08
Inventor 谢捷王平苏为科朱兴一王清荣杨明富
Owner ZHEJIANG UNIV OF TECH
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