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CN-121970895-A - High-activity eggshell membrane composite enzymolysis extraction process and extract

CN121970895ACN 121970895 ACN121970895 ACN 121970895ACN-121970895-A

Abstract

The invention belongs to the technical field of biological extraction, but is not limited to, and discloses a high-activity eggshell membrane compound enzymolysis extraction process and an extract, wherein the process adopts a compound enzyme system of alkaline protease and papain for directional hydrolysis, combines low-temperature ultrafiltration purification and freeze-drying technologies, the total amino acid content of the obtained extract is more than or equal to 79g/100g, the sodium hyaluronate content is more than or equal to 4.4g/100g, the collagen content is more than or equal to 6.8%, the water solubility is more than or equal to 95%, and the skin and eye irritation is avoided, so that the process can be widely applied to the fields of food health care, cosmetics, pet foods and the like, solves the technical problems of active ingredient loss and poor water solubility of the traditional process, and is stable and controllable and suitable for industrial production. The extract obtained by the invention has the total amino acid content of 79.3g/100g, the sodium hyaluronate content of 4.4g/100g and the collagen content of 6.8%, has no skin and eye irritation, can meet the application requirements of multiple fields such as food health care, cosmetics and the like, has mature and controllable process, and is suitable for industrialized mass production.

Inventors

  • YANG TAO
  • WANG YU
  • JIANG JUN

Assignees

  • 吉林生奥生物科技有限公司

Dates

Publication Date
20260505
Application Date
20260126

Claims (6)

  1. 1. A high-activity eggshell membrane composite enzymolysis extraction process is characterized by comprising the following steps: s1, raw material pretreatment, namely selecting traceable selenium-rich antibiotic-free eggshells, removing impurities, adopting a self-grinding high-efficiency shell membrane separation system, separating shell membranes under the conditions of 0.3-0.5MPa pressure and 10-15 ℃, collecting eggshell membranes, and crushing the eggshell membranes to 150-200 meshes to obtain eggshell membrane powder; s2, extracting at low temperature, mixing eggshell membrane powder with deionized water according to a feed-liquid ratio of 1:15-1:25, stirring and extracting at 30-35 ℃ for 1-2h, centrifuging at 8000-10000r/min for 15-20min, and taking supernatant to obtain crude extract; s3, composite enzymolysis, namely adding a composite enzyme preparation accounting for 1.5% -2.5% of the mass of eggshell membrane powder into the crude extract, adjusting the pH value to 7.0-7.5, and carrying out enzymolysis for 2.5-3.5 hours at 45-50 ℃ with continuous stirring at 100-120r/min in the enzymolysis process; S4, enzyme inactivation, namely heating the enzymolysis liquid to 85-90 ℃, preserving heat for 15-20min, inactivating enzyme activity, and cooling to room temperature; S5, gradient purification, namely purifying by adopting an ultrafiltration membrane with the molecular weight cut-off of 10000Da, collecting a permeate, and concentrating by nanofiltration until the solid content is 20-25% to obtain a purified concentrate; S6, drying, freeze-drying the purified concentrated solution at the temperature of-35 ℃ to-40 ℃ for 18-24 hours, and crushing to obtain the high-activity eggshell membrane extract.
  2. 2. The high-activity eggshell membrane composite enzymolysis extraction process according to claim 1, wherein the total enzyme activity of the composite enzyme preparation in the S3 is 100000-120000U/g, and nitrogen is introduced to protect the active ingredients in the enzymolysis process.
  3. 3. The high activity eggshell membrane composite enzymolysis extraction process according to claim 1, wherein the ultrafiltration operating pressure in S5 is 0.2-0.3MPa, and the nanofiltration operating pressure is 1.0-1.2MPa.
  4. 4. The high-activity eggshell membrane composite enzymolysis extraction process according to claim 1, wherein the eggshell membrane powder in S1 is required to meet the non-antibiotic and selenium-rich certification, and the selenium content is more than or equal to 0.15mg/kg.
  5. 5. A high activity eggshell membrane extract prepared by the process of any one of claims 1 to 4, wherein the total amount of amino acids of the extract is not less than 79g/100g, the sodium hyaluronate content is not less than 4.4g/100g, the collagen content is not less than 6.8%, the water solubility is not less than 95%, and the extract has no irritation to skin and eyes.
  6. 6. The high activity eggshell membrane extract of claim 5, wherein the extract is used as a food health raw material, a cosmetic raw material, a pet food functional additive or a nutrition enhancer in the feed industry.

Description

High-activity eggshell membrane composite enzymolysis extraction process and extract Technical Field The invention belongs to the technical field of biological extraction, but is not limited to, and particularly relates to a high-activity eggshell membrane composite enzymolysis extraction process and an extract. Background With the increase of comprehensive utilization of biological resources and development demands of high-added-value raw materials, eggshell membranes are used as a large amount of solid waste in the egg processing industry, and high-content collagen-like proteins, glycosaminoglycans and various bioactive substances of the eggshell membranes are gradually paid attention to the fields of foods, cosmetics and medicines. Eggshell membranes themselves have complex fibrous network structures that are difficult to separate intact and efficiently release their functional components by simple physical methods, thus creating a systematic technical development challenge. In the prior art, a typical scheme is to treat eggshell membranes or eggshell-shell lower membrane systems based on an enzymolysis method so as to extract collagen peptides or other active proteins. For example, for the extraction of eggshell membrane collagen, enzymolysis of a single enzyme is adopted, and conditions such as pH, enzyme dosage, feed-liquid ratio and the like are optimized through an orthogonal test, so that the recovery rate of specific components is improved. The scheme adopts papain to carry out enzymolysis at normal temperature under the conditions of pH5 and a feed-water ratio of 1:100, and a certain amount of collagen yield is obtained after long-time reaction. In another technical scheme, the separation efficiency is improved by means of composite enzymolysis or combination of chemical pretreatment and enzymolysis, for example, non-starch polysaccharase is used for hydrolyzing membrane surface fibers, and then protease is used for further acting on structural proteins, so that separation of eggshells and eggshell membranes is realized. Although the complex enzyme system can improve the effect of shell/membrane separation, the technology focuses on material separation per se, and does not make deep synergistic design for a system extraction process for extracting high-grade bioactive components such as high-purity collagen active peptide, polypeptide and other low-molecular active components. Although the schemes in the prior art can achieve partial targets, the obvious limitations still exist that firstly, most methods focus on the extraction of single components or the mechanical separation of membranes, an integral optimization flow from the separation of shell membranes to the release of low-molecular active components is not formed on the design of a process path, secondly, the penetration and compatibility of a single enzymolysis condition or a single enzyme system in a complex network structure of eggshell membranes are limited, so that the extraction efficiency of target components is low, the water solubility and purity of an extracted product are not high, thirdly, the inherent synergic mechanism between the enzymolysis process and the extracted components is not fully revealed in the prior art, and a process frame capable of continuously separating and purifying under the conditions of preserving heat-sensitive and structurally sensitive active substances is not constructed. These short plates are the core pain points which the present invention directly solves through the synergistic effect of low temperature water phase extraction and complex enzyme and molecular weight selective membrane separation integrated design. Disclosure of Invention Aiming at the problems existing in the prior art, the invention provides a high-activity eggshell membrane composite enzymolysis extraction process. The invention is realized in such a way that a high-activity eggshell membrane composite enzymolysis extraction process comprises the following steps: s1, raw material pretreatment, namely selecting traceable selenium-rich antibiotic-free eggshells, removing impurities, adopting a self-grinding high-efficiency shell membrane separation system, separating shell membranes under the conditions of 0.3-0.5MPa pressure and 10-15 ℃, collecting eggshell membranes, and crushing the eggshell membranes to 150-200 meshes to obtain eggshell membrane powder; s2, extracting at low temperature, mixing eggshell membrane powder with deionized water according to a feed-liquid ratio of 1:15-1:25, stirring and extracting at 30-35 ℃ for 1-2h, centrifuging at 8000-10000r/min for 15-20min, and taking supernatant to obtain crude extract; s3, composite enzymolysis, namely adding a composite enzyme preparation accounting for 1.5% -2.5% of the mass of eggshell membrane powder into the crude extract, adjusting the pH value to 7.0-7.5, and carrying out enzymolysis for 2.5-3.5 hours at 45-50 ℃ with continuous stirring at 100-120r/min in the enzym