Method for preparing Acaudina leucoprocta collagen oligopeptide through hydrolysis with combination of collagenase and novel microbial protease

A technology of microbial protease and collagenase, which is applied in the field of preparation of collagen peptides, can solve the problems of waste of collagen nutrients, difficult control of the process, incomplete hydrolysis, etc., and achieve the effects of increasing affinity, reducing pain, and shortening hydrolysis reaction time

Inactive Publication Date: 2018-05-18
ZHEJIANG OCEAN UNIV
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Problems solved by technology

[0005] The Chinese invention patent of CN 102251006 B discloses a method for producing collagen from black ginseng of the East China Sea. In the method, a complex protease is added to the serum of black ginseng of the East China Sea for direct hydrolysis, and then the collagen of the black ginseng of the East China Sea is prepared by membrane filtration, concentration, and drying. Protein products have the characteristics of a simple preparation method, but the process of this method is not easy to control, because the three-strand superhelical structure of collagen is relatively stable, so it is easy to cause incomplete hydrolysis, resulting in waste of collagen nutrients, and the preparation method has added Too much protease, because the enzymatic reaction rate has K M value, not by the amount of enzyme, thus causing waste of enzyme

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  • Method for preparing Acaudina leucoprocta collagen oligopeptide through hydrolysis with combination of collagenase and novel microbial protease

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Embodiment 1

[0022] A method for hydrolyzing black ginseng collagen oligopeptides in combination with novel microbial proteases, comprising the following steps:

[0023] 1) In the 0.04mol / L acetic acid solution system, add fresh and clean body wall homogenate of black ginseng with a solid-to-liquid ratio of 1:5, and then add pepsin with a substrate mass of 0.3-0.5% for restriction enzymolysis , the temperature was controlled at 5°C, the enzymolysis was carried out for 2 hours under continuous low-speed stirring, the enzymolysis solution was centrifuged at 12000r / min at high speed, the supernatant was taken, the precipitation was repeated for 3 times, the supernatant was collected, and freeze-dried to obtain Donghai Dried black ginseng collagen product; limited enzymatic hydrolysis in a weak acid system, which not only ensures the invariance of collagen, but also has a high collagen extraction rate, which can reach more than 50%;

[0024] 2) Prepare the fermentation medium as follows: glyce...

Embodiment 2

[0030] A method for preparing black ginseng collagen oligopeptides by combining collagenase with novel microbial protease hydrolysis, comprising: pretreatment, extraction of collagen, fermentation, purification of microbial protease, compound enzyme enzymolysis, microbial protease enzymolysis, and purification, specifically including the following step:

[0031] 1) Pretreatment: Clean the soaked black ginseng body wall, soak it with 5% hydrochloric acid solution and 0.08% calcium hydroxide solution for 36 hours, stir intermittently, change the solution every 12 hours, and use deionized water after soaking Rinse with water until neutral; then soak the sea cucumber body wall in 4% sodium hydroxide solution for 60 minutes, the sea cucumber body wall is transparent, rinse with deionized water until neutral, then chop and homogenize; pretreatment Calcium and non-collagen substances in the body wall of sea cucumbers can be removed to facilitate further extraction of collagen;

[00...

Embodiment 3

[0039] A method for preparing black ginseng collagen oligopeptides by combining collagenase with novel microbial protease hydrolysis, comprising: pretreatment, extraction of collagen, fermentation, purification of microbial protease, compound enzyme enzymolysis, microbial protease enzymolysis, and purification, specifically including the following step:

[0040] Pretreatment: Clean the soaked black ginseng body wall, soak in 4% hydrochloric acid for 24 hours, stir intermittently, change the solution every 10 hours, rinse with deionized water until neutral after soaking; then 0.06% hydroxide Calcium solution is also immersed in the same operation. After soaking, it is washed with deionized water until neutral; the body wall of sea cucumber is soaked in 4% sodium hydroxide solution for 50 minutes. The body wall of sea cucumber is transparent. Neutral, then chopped and homogenized; pretreatment can remove calcium and non-collagenous substances in the body wall of sea cucumbers, s...

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Abstract

The invention discloses a method for preparing Acaudina leucoprocta collagen oligopeptide through hydrolysis with combination of collagenase and novel microbial protease. The method comprises: pretreatment, collagen extraction, fermentation, microbial protease purification, enzymolysis, microbial protease enzymolysis, and purification. According to the present invention, during the enzymolysis, the dual-frequency ultrasonic wave assisted enzymolysis is used, such that the energy barrier between the ground state and the active state of the hydrolysis reaction can be reduced, and the Ea value and the KM value of the enzymolysis reaction can be reduced so as to increase the affinity between the matrix and the enzyme, greatly increase the enzymatic reaction rate, and save the time cost and thelabor cost; after the Acaudina leucoprocta collagen is hydrolyzed by combining the collagenase and the novel microbial protease, the collagen is hydrolyzed into the amino acids with low molecular weight and the oligopeptide with low molecular weight so as to be easily absorbed by the body and easily provide the medical health effects of the Acaudina leucoprocta collagen oligopeptide; and the obtained product is the nutrition health substance with characteristics of easy absorption, high nutrition and no toxic-side effect.

Description

technical field [0001] The invention relates to the field of preparation of collagen peptides, in particular to a method for hydrolyzing black cucumber collagen oligopeptides with collagenase combined with novel microbial proteases. technical background [0002] East China sea black ginseng (scientific name: white anus sea sweet potato Acaudina leucoprocta ), abounds in the East China Sea, and is widely distributed in our province, especially in the coastal areas of Ningbo and Zhoushan. Donghai black cucumber has extremely high nutritional value and medicinal value. It is rich in protein, sea cucumber saponin, sea cucumber polysaccharide, collagen, vitamins, essential fatty acids, and major and trace elements. It does not contain cholesterol. Long-term consumption can enhance physical fitness, Prevent diseases, inhibit tumors, and delay aging. The body wall of black ginseng is rich in collagen, which has good biological activity and biological function, and can participate...

Claims

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Application Information

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IPC IPC(8): C12P21/06
CPCC12P21/06
Inventor 金火喜张千伟杨立业欧阳小琨
Owner ZHEJIANG OCEAN UNIV
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