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Preparation method for active collagen from compound protease

A compound protease and collagen technology, applied in the field of collagen extraction, can solve problems such as difficult separation and purification, enzyme variability, and immature detection methods and technologies

Inactive Publication Date: 2014-10-01
林琳
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Problems solved by technology

For the biological activity of most active collagen, its industrial production is still difficult, mainly because the enzyme is easy to denature in the process of collagen hydrolysis, the separation and purification are difficult, and the detection methods and technologies are not mature enough.

Method used

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Embodiment Construction

[0013] Below in conjunction with specific embodiment, further illustrate the present invention, should be understood that these embodiments are only used to illustrate the present invention and are not intended to limit the scope of the present invention, after having read the present invention, those skilled in the art will understand various equivalent forms of the present invention All modifications fall within the scope defined by the appended claims of the present application.

[0014] A method for preparing active collagen by using a compound protease, in which 2.0-8.0% fish skin mixture is adjusted to a pH of 2-6 for pretreatment, and then a compound protease of papain, trypsin and pepsin is added. The content is the same as that of fish skin, and the temperature is controlled at 10-12°C, and the enzymatic hydrolysis is carried out for 8 hours. During this process, the pH is controlled between 5-9. Then the supernatant after enzymolysis is centrifuged at about 4500r / min...

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Abstract

The invention discloses a preparation method for active collagen from compound protease. The method comprises the following steps: 1) adding compound protease into pretreated fish skin mixed liquor for enzymatic hydrolysis, wherein the compound protease at least comprises two selected from the group consisting of papain, bromelain, trypsin, neutral protease, animal protein hydrolase, acidic protease and pepsin; 2) subjecting the obtained enzymatic hydrolysis liquid to centrifugation and taking supernatant; 3) subjecting the supernatant to ultrafiltration and concentration and then to salting-out; and 4) carrying out ultrafiltration, desalination and concentration; wherein the fish skin mixed liquor has a concentration of 2.0 to 8.0 wt% and a pH value of 2 to 6, and the addition amount of the compound protease is 2.0 to 8.0 wt% of the weight of the fish skin mixed liquor. The method provided by the invention has high security and no side-effects; and the prepared active collagen has the advantages of a high extraction rate, high purity, high thermal denaturation temperature and the like.

Description

[0001] technical field [0002] The invention belongs to the field of biochemistry, in particular to a method for extracting collagen. Background technique [0003] At present, domestic research on the use of compound enzymes to extract active collagen has just started. Several domestic universities have carried out research on compound enzymes to extract active collagen, and have made certain progress in the selection of proteases and enzymatic hydrolysis processes. At present, active collagen is extracted by enzymatic hydrolysis, and the proteolytic enzymes used are relatively single, mainly trypsin, pepsin, alkaline protease, neutral protease, papain, etc. For the biological activity of most active collagen, its industrial production is still difficult, mainly because the enzyme is easy to denature during the hydrolysis process of collagen, the separation and purification are difficult, and the detection means and technology are not mature enough. Contents of the invent...

Claims

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Application Information

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IPC IPC(8): C12P21/06
Inventor 程张军郁帅陆黄荣醒陈美娟
Owner 林琳
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