CN116875572A - Method for extracting superoxide dismutase - Google Patents
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- CN116875572A CN116875572A CN202311038405.XA CN202311038405A CN116875572A CN 116875572 A CN116875572 A CN 116875572A CN 202311038405 A CN202311038405 A CN 202311038405A CN 116875572 A CN116875572 A CN 116875572A
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0089—Oxidoreductases (1.) acting on superoxide as acceptor (1.15)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y115/00—Oxidoreductases acting on superoxide as acceptor (1.15)
- C12Y115/01—Oxidoreductases acting on superoxide as acceptor (1.15) with NAD or NADP as acceptor (1.15.1)
- C12Y115/01001—Superoxide dismutase (1.15.1.1)
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
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- Coloring Foods And Improving Nutritive Qualities (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
The application relates to a method for extracting superoxide dismutase, which comprises a crushing and juicing process, a solid-liquid separation process, an extraction and concentration process and an immobilization modification process; the plant SOD product is produced by a biphasic enzyme method extraction and enzyme immobilization modification technology, wherein the immobilization modification technology is to prepare immobilized SOD by an immobilization optimization technology through crude SOD enzyme, and then further purify and modify the immobilized SOD to prepare the raw material SOD required by high-activity SOD.
Description
Technical Field
The application relates to the technical field of biology, in particular to a method for extracting superoxide dismutase.
Background
Superoxide dismutase (SOD) is one of the most important antioxidant enzymes of the metabolic and defensive system in the human body, and its role is invisible. Is a natural scavenger of oxygen free radicals in organisms, has wide medical value, and can be used as an additive of medicines, foods and daily chemical products. Can be used for treating various diseases caused by abnormal increase of superoxide radical, such as osteoarthritis, rheumatoid arthritis, side effects caused by radiotherapy and chemotherapy, brain injury, reperfusion syndrome, etc. Therefore, the enzyme is praised as 'enzyme with magic' by the world science and technology world, is a natural gram of radical source of ten kinds of diseases, is a specific scavenger, is a protecting spirit of human life, is a 'scavenger' of human garbage, is a longevity factor for prolonging life, and is a cosmetic proud for delaying aging; is the only substance for reversing the biological clock of human beings, and is the reputation of gold keys and the like for the health and longevity of human beings and the returning of children.
The existing superoxide dismutase (SOD) is mainly prepared from animals, plants and microorganisms. The majority of the superoxide dismutase (SOD) products currently on the market are extracted from animal blood. Due to limited raw materials, difficult purification and the like, superoxide dismutase (SOD) has low purity and low yield. At present, the enzyme activity and specific activity of superoxide dismutase (SOD) extracted from plants are difficult to break through tens of millions of international units.
Disclosure of Invention
The application aims to provide a method for extracting superoxide dismutase, which is an advanced technology of modern bioengineering with the characteristics of safe and environment-friendly process, high productivity, high efficiency, good quality and the like. The method can only meet the requirement of extracting SOD enzyme from single raw material and single process in the same industry, but the technical process can be suitable for extracting SOD enzyme from various plants, solves the problems of product application safety and national medical and edible standard, and also solves the defects and application problems of incapability of eating and medical graded production in the same industry production process.
In order to achieve the above object, on the one hand, the present application relates to the following technical solutions: a method for extracting superoxide dismutase is technically characterized by comprising the following steps:
the method for extracting superoxide dismutase comprises a crushing and juicing process, a solid-liquid separation process, an extraction and concentration process and an immobilization modification process;
the crushing and juicing process comprises the steps of firstly cutting, cleaning and preprocessing raw materials, and then soaking so as to realize pulping and juicing;
the solid-liquid separation process comprises the steps of carrying out coarse filtration, impurity removal, decolorization and sterilization after a fixed separation step by low-temperature heating, carrying out fine filtration step by temperature adjustment and PH value adjustment, and carrying out heating precipitation on SOD enzyme solution;
the extraction and concentration process is to perform chromatographic separation on the heated and precipitated total enzyme solution, namely a double-effect concentration step, so that SOD crude enzyme, namely a plant leaf protein solution, can be prepared;
the immobilized modification process is to obtain immobilized SOD by the immobilized optimization technology of the crude enzyme of the SOD, and then further purify and modify the immobilized SOD to obtain the high-activity SOD.
Preferably, the specific processing steps of the method for extracting superoxide dismutase are as follows:
s1: fresh plant stems or leaves from base picking are carefully selected and cleaned, crushed by special equipment until the process requirements are met, and sent into a juice squeezing wall breaking system to obtain plant slurry and fresh leaf residues, wherein the plant slurry is used for SOD crude enzyme and SOD enzyme liquid, and the residues are used for producing feed;
s2: the plant slurry obtained in the step S1 is subjected to solid-liquid separation, and the steps comprise temperature adjustment, pH value adjustment, coarse and fine filtration, salting-out precipitation and separation to obtain SOD crude enzyme and SOD enzyme liquid, and the plant leaf protein is used as a new food raw material additive;
s3: concentrating the SOD enzyme solution prepared in the step S2, performing reverse phase chromatography to obtain SOD crude enzyme, preparing immobilized SOD by the immobilized optimization technology, and further purifying and modifying to obtain high-activity SOD.
The application has the advantages and beneficial effects that: the method is to prepare a plant SOD product by a biphasic enzyme method extraction and enzyme immobilization modification technology, the plant SOD product is subjected to the detection of the enzyme activity reaching tens of thousands of international units (1936.22) by a China Guangzhou analysis test center of a third-party authority detection mechanism, the specific activity is purified to reach 2900 tens of thousands of international units (2.9X107 u/g. Protein), the enzyme activity and the specific activity purity of the product exceed those of a American Sigma standard sample tens of thousands of international units (3.0X106 u/g. The protease activity is 2.6 times, the specific activity of the protein is 9.7 times, the product error of the same batch is less than 3 percent, and the product exceeds the international standard, so that the national blank is filled
Drawings
FIG. 1 is a schematic flow chart of a method for extracting superoxide dismutase according to the present application;
Detailed Description
In the present specification, each embodiment is described in a progressive manner, and each embodiment is mainly described in a different point from other embodiments, and identical and similar parts between the embodiments are all enough to refer to each other.
As shown in figure 1, the extraction process of superoxide dismutase (SOD) comprises a crushing and juicing process, a solid-liquid separation process, an extraction concentration process and an immobilization modification process of fresh plant leaves, and the related technologies comprise a heating and solidification leaf protein technology, an acid precipitation leaf protein technology, a reversed phase chromatography separation technology, an immobilized enzyme technology and a modification technology.
Firstly, carefully selecting and cleaning fresh plant stems or leaves from a base, crushing by special equipment until the plant stems or leaves meet the technological requirements, and feeding the plant stems or leaves into a juice-squeezing wall-breaking system to obtain plant pulp and fresh leaf residues, wherein the plant pulp is used for extracting plant leaf proteins and SOD, and the residues are used for producing feed;
secondly, the plant slurry is subjected to solid-liquid separation, and the steps comprise temperature adjustment, namely temperature control, PH value adjustment, coarse and fine filtration, salting-out precipitation and separation by a heating or cooling method, so as to obtain plant leaf protein and SOD enzyme liquid, wherein the plant leaf protein is used as a new food raw material additive;
finally, the SOD enzyme solution is concentrated and subjected to reverse phase chromatography to obtain SOD crude enzyme, the SOD crude enzyme is prepared into immobilized SOD by an immobilization optimization technology, and then the immobilized SOD crude enzyme is further purified and modified to obtain high-activity SOD, and the industrial scale annual production of millions-grade SOD freeze-dried powder of 2 tons and tens of millions-grade medicinal enzyme SOD freeze-dried powder of 200 kilograms can be realized by the method for extracting superoxide dismutase. The plant leaf residues are used for producing nutritional feed, the plant leaf proteins are used for producing protein powder, dietary supplement products or new food raw materials for sale, and the SOD is used for producing SOD health food capsules, SOD tablets, SOD oral liquids and other food and health product additives, so that the full utilization of resources is realized. The obtained product mainly comprises superoxide dismutase (SOD), leaf protein, crude protein and the like, and can be widely applied to the fields of medicine, chemical industry, food and cosmetics, military industry, modern agriculture and the like.
Superoxide dismutase (SOD) is approved for clinical use and shows a strong regenerative repair capacity for some tissue hardening and fibrosis due to age, disease or injury. Superoxide dismutase (SOD) has been successfully used in the adjuvant therapy following radiotherapy, to control the progression of heart patients, and to treat severe rheumatoid arthritis.
Superoxide dismutase (SOD) medicinal enzymes are used in the pharmaceutical industry: in European and American countries, it has been approved as a clinically pharmaceutically acceptable enzyme. Superoxide dismutase (SOD) is used as a medicinal enzyme preparation, and has wide medical value and clinical application prospect. Superoxide dismutase (SOD) is favored in many countries such as europe and america because it has a good effect on the prevention and treatment of difficult and complicated diseases such as cancer. Superoxide dismutase (SOD) is used as a medicinal preparation, can resist radiation, treat burn, scald, accelerate wound healing and other various external medical raw materials for skin, has been developed on the market at present, and can also be used for inhibiting diseases such as tumor, coronary heart disease, prostatitis, gastric cancer and the like. The expansion of superoxide dismutase (SOD) in the field of medical applications depends on the more intensive research on its mechanism of action and mechanism of production, and the discovery of the pro-factor of superoxide dismutase (SOD) is expected.
Superoxide dismutase (SOD) food industry application: SOD is used as a nutrient supplement for health foods, and people want to keep SOD at a high level in the human body by using various feasible methods, namely, providing proper nutrition for the liver to synthesize SOD while consuming food, or directly wrapping exogenous SOD in food protein, passing through the digestive system without being digested, and entering other positions of the body. Along with the development of science and technology, the research and development of a fourth-generation health care product containing SOD reveals mystery veil, and the SOD health care product starts from the real root source for solving the human aging and diseases, adjusts the physiological balance of human body, and is consistently considered by the scientific and technological community, medical community and food specialist: is the health food which is most beneficial to the health of human body at present. Such as: SOD can be widely used as an antioxidant nutrient health-care food functional factor in the food industry. The SOD can be added into food to enhance health promotion effects such as antiaging, antiinflammatory, radioprotective, and antifatigue effects. Can also be used as antioxidant in canned food, fruit juice, beer, etc., to prevent food deterioration and putrefaction caused by peroxidase; can also be used as antistaling agent for fruits and vegetables.
Superoxide dismutase (SOD) is used in the cosmetic industry: there are hundreds of products in cosmetics such as SOD mask, SOD honey and SOD snake powder developed by using superoxide dismutase at home. The federal food administration calls superoxide dismutase (SOD) to be: clinical experiments prove that the SOD beauty skin care product has 88.6 percent of effective rate on removing wrinkles and 87.8 percent of effective rate on removing freckles. The effective rate of removing acnes reaches 94.7 percent. The effective rate of removing pigmentation reaches 91.8 percent, and the total effective rate of SOD cosmetic and beauty product is 90.17 percent; the application of superoxide dismutase (SOD) in cosmetics can be used as raw material of cosmetics for whitening skin, resisting aging, resisting oxidation and resisting allergy. Research has shown that the addition of superoxide dismutase (SOD) to cosmetics can play four roles:
firstly, the sun-proof effect is obvious, the main reason for blackening the skin by illumination is oxygen radical damage, and superoxide dismutase (SOD) can effectively prevent the skin from being damaged by ionizing radiation (particularly ultraviolet rays), so that the sun-proof effect is achieved;
secondly, superoxide dismutase (SOD) is antioxidant enzyme, and can effectively prevent skin aging, remove spots and resist wrinkles;
thirdly, the traditional Chinese medicine composition has obvious anti-inflammatory effect and a certain effect on preventing and treating skin diseases; fourthly, superoxide dismutase (SOD) has certain effect of preventing and treating the formation of the speckles.
Superoxide dismutase (SOD) is used in agriculture: SOD inducer and plant protection agent for green plants, the physicochemical effects of which are two:
firstly, can increase the SOD content in plants. The SOD enzyme agent sprayed on the plant leaf surface can promote the differentiation of leaf green cell tissue to generate more SOD enzyme after being absorbed by plant body and stored in mesophyll cells of green plant. Through scientific detection, the SOD content is 3-5 times of that of the original plant, so that the plant with low SOD content can be changed into the plant with high SOD content, and the common plant food can be changed into the plant food with health care function.
And secondly, the stress resistance and the disease and pest resistance of plant varieties can be improved. The leaf protein can be used as new food material, and the plant leaf residue can be used for producing nutritional feed. Along with the improvement of the living standard of people, the nutrition food with high content of superoxide dismutase (SOD) is an essential food therapy good product for people to seek medical and edible science and root, so that the superoxide dismutase (SOD) industry is greatly developed, the future world superoxide dismutase (SOD) biotechnology is high, the cluster of the superoxide dismutase (SOD) biomedical food industry is extremely created, the 21 st century biological economy is met, the contribution is made to the health and longevity industry of people, and the nutrition food with high content of superoxide dismutase (SOD) is also a great strategic significance and has a great unpredictable market demand broad prospect for the development of green industry and the age of the great health industry.
The method for extracting superoxide dismutase adopted by the application adopts an enzyme method in the extraction process, and comprises the following steps:
the previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present application. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the application. Thus, the present application is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (2)
1. A method for extracting superoxide dismutase, which is characterized by comprising the following steps:
the method for extracting superoxide dismutase comprises a crushing and juicing process, a solid-liquid separation process, an extraction and concentration process and an immobilization modification process;
the crushing and juicing process comprises the steps of firstly cutting, cleaning and preprocessing raw materials, and then soaking so as to realize pulping and juicing;
the solid-liquid separation process comprises the steps of carrying out coarse filtration, impurity removal, decolorization and sterilization after a fixed separation step by low-temperature heating, carrying out fine filtration step by temperature adjustment and PH value adjustment, and carrying out heating precipitation on SOD enzyme solution;
the extraction and concentration process is to perform chromatographic separation on the heated and precipitated total enzyme solution, namely a double-effect concentration step, so that SOD crude enzyme, namely a plant leaf protein solution, can be prepared;
the immobilized modification process is to obtain immobilized SOD by the immobilized optimization technology of the crude enzyme of the SOD, and then further purify and modify the immobilized SOD to obtain the high-activity SOD.
2. The method for extracting superoxide dismutase according to claim 1, wherein: the method comprises the following steps:
s1: fresh plant stems or leaves from base picking are carefully selected and cleaned, crushed by special equipment until the process requirements are met, and sent into a juice squeezing wall breaking system to obtain plant slurry and fresh leaf residues, wherein the plant slurry is used for SOD crude enzyme and SOD enzyme liquid, and the residues are used for producing feed;
s2: the plant slurry obtained in the step S1 is subjected to solid-liquid separation, and the steps comprise temperature adjustment, pH value adjustment, coarse and fine filtration, salting-out precipitation and separation to obtain SOD crude enzyme and SOD enzyme liquid, and the plant leaf protein is used as a new food raw material additive;
s3: concentrating the SOD enzyme solution prepared in the step S2, performing reverse phase chromatography to obtain SOD crude enzyme, preparing immobilized SOD by the immobilized optimization technology, and further purifying and modifying to obtain high-activity SOD.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202311038405.XA CN116875572A (en) | 2023-08-17 | 2023-08-17 | Method for extracting superoxide dismutase |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202311038405.XA CN116875572A (en) | 2023-08-17 | 2023-08-17 | Method for extracting superoxide dismutase |
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| CN116875572A true CN116875572A (en) | 2023-10-13 |
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