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Biocatalytic process for preparing collagen peptide

A collagen peptide and biocatalysis technology, applied in the field of biocatalysis and biosynthesis, can solve the problems of many steps, environmental pollution, low hydrolysis rate, etc., and achieve simplified product separation steps, simple and controllable reaction process, and mild reaction conditions. Effect

Active Publication Date: 2007-06-06
珠海凤凰高科生物制药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods generally have the following disadvantages at present: (1) many steps, low hydrolysis rate, high energy consumption, and difficult separation of products; (3) After the reaction, high-temperature inactivation of enzymes is often used to stop the reaction, which not only increases energy consumption, but also makes it difficult to separate and remove enzymes and enzymes after inactivation. The low utilization rate greatly increases the economic cost of the method and the difficulty of product separation and purification; (4) the discharge of a large amount of alkali waste liquid after the reaction will bring serious environmental pollution problems

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Put 0.5g of collagen powder, 0.5mg of immobilized protease derived from papaya latex and 50ml of water / butane (100:1) two-phase medium into a stoppered Erlenmeyer flask, and place it in a water bath constant temperature oscillator at 30°C and 100rpm Internal shaking, after reacting for 3 hours, the immobilized protease was removed by filtration, the organic phase was removed after centrifugation, and the remaining solution was subjected to vacuum rotary evaporation at 50°C, and the obtained precipitate was fractionated with ultrafiltration membrane 6000 membrane and 5000 membrane to obtain 6000~5000Da collagen peptides.

Embodiment 2

[0020] Put 0.5g of collagen powder, 0.5mg of immobilized lipase derived from Bacillus lichemiforms and 50ml of water / octanol (1:1) biphasic medium into a stoppered Erlenmeyer flask, place it in a water bath at 60°C and 200rpm to shake at a constant temperature Shake in the vessel, react for 12 hours, remove the immobilized protease by filtration, remove the organic phase after centrifugation, and carry out vacuum rotary evaporation at 50°C for the remaining solution, and grade the obtained precipitates with ultrafiltration membranes 5000 and 2000 with different cut-offs Separation can obtain collagen peptides with a molecular weight of 5000-2000Da.

Embodiment 3

[0022] Put 0.5g of collagen powder, 5mg of immobilized protease derived from Bacillus amyloliquefaciens and 10ml of water / toluene (100:1) two-phase medium into a stoppered Erlenmeyer flask, and shake in a water bath constant temperature oscillator at 50°C and 300rpm , after reacting for 0.5h, remove the immobilized protease by filtration, remove the organic phase after centrifugation, and carry out vacuum rotary evaporation at 70°C for the remaining solution, and fractionate the obtained precipitate with ultrafiltration membrane 2000 membrane and 1000 membrane to obtain a molecular weight of 2000 Collagen peptides of ~1000 Da.

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Abstract

The biocatalytic process for prepare collagen peptide includes the following steps: compounding double phase reaction medium with organic solvent and water in the volume ratio of 0.01-1 to 1, adding collagen powder in the collagen / reaction medium ratio of 0.01-10 g to 1 ml, adding immobilized hydrolase in the biocatalyst / collagen powder weight ratio of 0.001-10 to 1, reaction at 50-70 deg.c, vibration speed of 100-300 rpm and normal pressure for 0.5-48 hr, and separation to obtain collagen peptide. The process has the advantages of low cost, mild reaction condition, environment friendship, simple and controllable reaction, and easy product separation.

Description

technical field [0001] The invention relates to the fields of biocatalysis and biosynthesis, in particular to a method for preparing collagen peptides by biocatalysis in a water / organic solvent two-phase system. Background technique [0002] Collagen is the main component of animal connective tissue. It is the most abundant and widely distributed protein with immunological properties in animals. It is particularly abundant in animal skin tissue and bones. For example, the protein content of pigskin is as high as 26.4%, of which collagen accounts for 87.8%; rough collagen in fish scales and fish skin accounts for about 80%; animal bones are also rich in this kind of protein, accounting for about 70.0% of organic matter -80.0%. However, for a long time, these collagen-rich animal tissues were mostly discarded as industrial waste, or their utilization rate was extremely low. For example, the bones obtained after animal product processing are often made into bone powder and so...

Claims

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Application Information

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IPC IPC(8): C12P21/02C12R1/01C12R1/10C12R1/125C12R1/41C12R1/645C12R1/73
Inventor 吴晖李晓凤余以刚刘冬梅许喜林
Owner 珠海凤凰高科生物制药有限公司