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A method for preparing biologically active substances from eggshell membranes by a compound enzyme hydrolysis method

A bioactive substance and compound enzyme hydrolysis technology, applied in the field of bioengineering, can solve the problems of severe reaction, large loss of active substances, low enzymatic hydrolysis efficiency, etc., achieve mild reaction conditions, easy process control, and improve enzymatic hydrolysis efficiency Effect

Active Publication Date: 2022-05-27
SHANGHAI FOREST CABIN BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional eggshell membrane hydrolysis process is mainly based on acid and alkali hydrolysis, which has the advantages of simplicity, convenience, and easy implementation. However, it is easy to produce toxic and harmful substances to the environment during the production process, which makes post-processing of the product difficult. The reaction is violent and difficult to control during the hydrolysis process, which causes serious damage to the molecular structure of the protein and a large loss of active substances.
The enzymatic hydrolysis process belongs to the biological method, which has the advantages of mild reaction conditions, easy control of the process, environmental friendliness, low purification cost, and stable enzymatic hydrolysis products. Therefore, it is particularly important to improve the efficiency of enzymatic hydrolysis by pretreatment of eggshell membranes. For example, a method for preparing eggshell membrane polypeptides with a domestic application number of 201911080199.2 and a method for preparing eggshell membrane polypeptides with a domestic application number of 201410521600.2 The method of extracting biologically active substances in the shell membrane adopts the pretreatment method of alkaline solution and ionic liquid to improve the efficiency of enzymatic hydrolysis, but the use of these harmful reagents reduces the technological advantages of enzymatic hydrolysis

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  • A method for preparing biologically active substances from eggshell membranes by a compound enzyme hydrolysis method
  • A method for preparing biologically active substances from eggshell membranes by a compound enzyme hydrolysis method
  • A method for preparing biologically active substances from eggshell membranes by a compound enzyme hydrolysis method

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Embodiment 1

[0028] The present embodiment relates to a method for preparing biologically active substances from eggshell membranes by a compound enzymatic hydrolysis method, comprising the following steps:

[0029] (1) Preparation of eggshell membrane: The eggshell is separated by a shell membrane separator to achieve shell membrane separation, and the obtained eggshell membrane is cleaned, sterilized, dried at 80°C for 10 hours, then pulverized, and finally passed through a 40-mesh sieve net to obtain eggshell membrane powder;

[0030] (2) Pre-hydrolysis: mix the eggshell membrane powder with 10 times its dry weight of water, then heat the aqueous solution to 60°C, and stir at this temperature for 0.5h;

[0031] (3) Hydrolysis: reduce the aqueous solution in step (2) to 35°C, adjust the pH to 6.5, and then add 1% composite enzyme (the composite enzyme is composed of trypsin and neutral protease in a mass ratio of 1:4) , stirred and hydrolyzed for 3h;

[0032] (4) Preparation of biologi...

Embodiment 2

[0034] The present embodiment relates to a method for preparing biologically active substances from eggshell membranes by a compound enzymatic hydrolysis method, comprising the following steps:

[0035] (1) Preparation of eggshell membranes: The eggshells are separated by a shell membrane separator to achieve shell membrane separation, and the obtained eggshell membranes are cleaned and sterilized, dried at 80°C for 10 hours, then pulverized, and finally passed through an 80-mesh sieve. net to obtain eggshell membrane powder;

[0036] (2) Pre-hydrolysis: mix the eggshell membrane powder with 50 times of its dry weight water, then heat the aqueous solution to 80°C, and stir at this temperature for 2h;

[0037] (3) Hydrolysis: reduce the aqueous solution in step (2) to 60°C, adjust the pH to 9.5, and then add 10% composite enzyme (the composite enzyme is composed of trypsin and neutral protease in a mass ratio of 5:1) , stirred and hydrolyzed for 8h;

[0038] (4) Preparation o...

Embodiment 3

[0040] The present embodiment relates to a method for preparing biologically active substances from eggshell membranes by a compound enzymatic hydrolysis method, comprising the following steps:

[0041] (1) Preparation of eggshell membrane: The eggshell is separated by a shell membrane separator to achieve shell membrane separation, the obtained eggshell membrane is cleaned, sterilized, dried at 60°C for 8 hours, then pulverized, and finally passed through a 60-mesh sieve net to obtain eggshell membrane powder;

[0042] (2) Pre-hydrolysis: mix the eggshell membrane powder with water 25 times its dry weight, then heat the aqueous solution to 65°C, and stir at this temperature for 1 hour;

[0043] (3) Hydrolysis: reduce the aqueous solution in step (2) to 55°C, adjust the pH to 7.5, and then add 8% composite enzyme (the composite enzyme is composed of trypsin and neutral protease in a mass ratio of 3:2) , stirred and hydrolyzed for 6.5h;

[0044] (4) Preparation of biologicall...

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Abstract

The invention discloses a method for preparing biologically active substances from eggshell membranes by a compound enzyme hydrolysis method, which belongs to the technical field of bioengineering and mainly includes the following steps: preparation of eggshell membranes, prehydrolysis, multi-enzyme hydrolysis and preparation of biologically active substances Preparation, wherein the composite enzyme includes trypsin and neutral protease with a mass ratio of 1:4 to 5:1; the preparation method avoids the use of toxic reagents such as ionic liquids and organic solvents, and the use of the composite enzyme makes eggshell membrane hydrolysis easier Thoroughly, the resulting substance is rich in protein, hyaluronic acid, chondroitin sulfate and other active substances.

Description

technical field [0001] The invention relates to the technical field of bioengineering, and relates to a method for preparing biologically active substances from eggshell membranes by a compound enzymatic hydrolysis method; in particular, it relates to a method for improving enzymatic hydrolysis efficiency by pre-hydrolysis and multi-enzyme compounding. Methods for the preparation of biologically active substances. Background technique [0002] The eggshell membrane is mainly composed of crude protein, fat and polysaccharide, among which crude protein accounts for about 90% of the total weight of the membrane, mainly in the form of glycoprotein, and also contains about 3% of fat and 2% of polysaccharide. The protein is mainly keratin, and also contains collagen, lysozyme, ovotransferrin, ovalbumin, etc. The mucopolysaccharide is mainly composed of hyaluronic acid and chondroitin sulfate; It is widely used in food, cosmetic, pharmaceutical, chemical and other industries. The...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08B37/08
CPCC08B37/0003C08B37/0069C08B37/0072Y02P20/54
Inventor 李广涛奕志英秦枭高宏旗
Owner SHANGHAI FOREST CABIN BIOLOGICAL TECH