Application of immobilized proteinase in preparing cream protein peptide

A technology for immobilizing protease and milk casein, applied in the preparation of milk casein peptide, immobilized protease, immobilized enzyme and its application fields, can solve the problems that the research of milk casein peptide has not yet been reported, etc., and achieves convenient product separation, The effect of high peptide content and reduced production cost

Inactive Publication Date: 2010-07-07
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there have been researches on the preparation of milk casein peptides by hydrolyzing milk casein with protease in China, but there is no report on the preparation of milk casein peptides by hydrolysis of immobilized enzymes

Method used

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  • Application of immobilized proteinase in preparing cream protein peptide
  • Application of immobilized proteinase in preparing cream protein peptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] Example 1 Preparation of immobilized trypsin

[0081] (1) Stir 10% chitosan acid sol to disperse the sol droplets uniformly, and drop into 2.5% glutaraldehyde solution to make the final concentration (volume fraction) of glutaraldehyde solution reach 3% , Stir and add 3mol / L NaOH solution until the final concentration (volume fraction) of NaOH is 3%. Incubate at 70°C for 0.5% reaction. After standing and cooling, discard the upper layer, wash with acetone, absolute ethanol, and finally with distilled water Rinse and dry in vacuum to obtain immobilized carrier chitosan microspheres for use;

[0082] (2) Add immobilized carrier chitosan microspheres to 5% trypsin solution, where trypsin solution: immobilized carrier chitosan microspheres = 4:1; place it in a shaker shaker for 30 hours, then Dropping a 2.5% glutaraldehyde solution with a volume fraction to make the final concentration (volume fraction) of the glutaraldehyde solution reach 3%, and cross-linking for 0.2-10h to o...

Embodiment 2

[0083] Example 2 Preparation of immobilized pepsin

[0084] (1) Stir 0.5% chitosan acidic sol to uniformly disperse the sol droplets, and drop into 2.5% glutaraldehyde solution to make the final concentration (volume fraction) of the glutaraldehyde solution reach 0.2% , Add 0.5mol / L NaOH solution with stirring until the final concentration (volume fraction) of NaOH is 0.05%, keep at 40℃ for 5h, let stand and cool, discard the upper layer, wash with acetone and absolute ethanol in turn, and finally use Rinse with distilled water and dry in vacuum to obtain immobilized carrier chitosan microspheres for use;

[0085] (2) Add the prepared immobilized carrier chitosan microspheres to 0.1% pepsin solution, where pepsin solution: immobilized carrier chitosan microspheres = 1:1; placed in a shaker shaker for adsorption 2h, then drop in a 2.5% glutaraldehyde solution with a volume fraction to make the final concentration (volume fraction) of the glutaraldehyde solution reach 0.2%, and cros...

Embodiment 3

[0086] Example 3 Preparation of immobilized bromelain

[0087] (1) Stir 5% chitosan acidic sol to make the sol droplets evenly dispersed, and drop into 2.5% glutaraldehyde solution to make the final concentration (volume fraction) of the glutaraldehyde solution reach 0.5% , Add 0.8mol / L NaOH solution with stirring until the final concentration (volume fraction) of NaOH is 0.08%, keep at 50℃ and react for 2.5h, let it stand and cool, discard the upper layer, wash with acetone and absolute ethanol in turn, and finally Rinse with distilled water and dry in vacuum to obtain immobilized carrier chitosan microspheres for use;

[0088] (2) Add the prepared immobilized carrier chitosan microspheres accounting for 5% of the volume of the bromelain solution to the 1.5% bromelain solution, wherein the bromelain solution: immobilized carrier chitosan microspheres=3.5:1; Place it in a shaker shaker for 10 hours, then drop in a 2.5% glutaraldehyde solution to make the final concentration (volum...

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Abstract

The invention discloses an immobilized protease and an application method of the immobilized protease in preparation of milk casein peptides. The main preparation method of the immobilized protease is as follows: firstly, immobilized carrier chitosan microspheres are prepared; secondly, the chitosan microspheres are added into protease solution; after oscillation and absorption, glutaraldehyde solution is dripped into the solution; after cross linking, the immobilized protease is obtained. The method for preparation of the milk casein peptides adopts the immobilized protease to hydrolyze the milk caseins. The immobilized protease can be reused for a plurality of times, thereby the use efficiency is high and the production cost is effectively reduced; moreover, the immobilized protease hasthe advantages of convenient product separation, low cost, mild production conditions and easy control of the hydrolysis degree. The milk casein peptide product prepared has the advantages of high peptide content, superior peptide molecule weight distribution range, good quality, no bitterness, high safety and so on. The content (according to the dry basis) of the milk casein peptides prepared bythe method is as high as 75.05 percent by detection; the peptide molecule weight distribution range is that: the molecular weight which is lower than 2409 daltons reaches 97.79 percent.

Description

Technical field [0001] The invention relates to an immobilized enzyme and its application, in particular to an immobilized protease. The invention also relates to the application of the immobilized protease in the preparation of casein peptides, belonging to the technical field of food biology. Background technique [0002] Functional food is the focus of the development of the food industry in the 21st century. The development of various physiologically active functional nutritional factors has far-reaching biological significance and important application value. It is also the most popular research topic in the food industry at home and abroad and an industry with great development prospects. [0003] Milk contains a large number of biologically active peptides with different activities. These peptides are either naturally present in milk or derived from the products of milk protein hydrolysis. Because of its small molecular weight, it is absorbed by the human body in the form o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/04C12P21/06
Inventor 周利根郜海燕陈新峰王君虹
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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