Liquid superoxide-rich disproportionated active enzyme bred by sugar soaking of rosa roxburghii fruits after strong freezing and unfreezing
By artificially planting prickly pear fruits and using strong freeze-thawed sugar-grown liquid dosage form, the extraction efficiency of SOD active enzymes was successfully improved, the problems of low SOD content and high extraction cost in the prior art were solved, and efficient and low-cost SOD extraction was achieved.
Patent Information
- Application Number
- CN202510006231.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-01-03
- Publication Date
- 2025-03-18
AI Technical Summary
In the prior art, when extracting superoxide dismutase (SOD), the SOD content in the plants is low, resulting in complex extraction process and high cost.
Artificially planted prickly pear fruits are used to grow liquid dosage forms through strong freeze-thawing and sugar-growing, sugar-soaking is used for sugar-soaking, and then juice rich in SOD-active enzymes is obtained using heavy objects.
This method can breed SOD in a larger amount, reduce production costs, improve the absorption efficiency of SOD-active enzymes, and solve the technical problem that fresh fruits cannot be kept for a long time.
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Figure CN119632156A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of oxidoreductase, and specifically relates to a method for preparing superoxide dismutase by using plant fruits, and the obtained product. Background Art
[0002] Superoxide dismutase (abbreviated as SOD) is an enzyme that can catalyze the conversion of superoxide into oxygen and hydrogen peroxide through dismutation reaction. SOD is an important antioxidant that can remove various reactive oxygen species produced by cell metabolism and protect cells from oxygen free radicals. Superoxide dismutase exists in several common forms: they use copper and zinc, or manganese, iron, or nickel as cofactors. Basically all eukaryotic cells contain superoxide dismutase with copper and zinc (Cu-Zn-SOD); almost all mitochondria and many bacteria (such as Escherichia coli) contain superoxide dismutase bound to manganese (Mn-SOD). Escherichia coli and some other bacteria also contain superoxide dismutase bound to iron (Fe-SOD). In higher plants, different forms of superoxide dismutase are localized in different cell compartments. Mn-SOD exists in mitochondria and peroxisomes; Fe-SOD is mainly located in chloroplasts, but can also be detected in peroxisomes; CuZn-SOD is located in protoplasm, chloroplasts, peroxisomes and apoplasts, so it is feasible to extract SOD from plant tissues. There are many plants that can be used to extract SOD, including sea buckthorn, garlic, cactus, etc., but the SOD content in most plants is low, which makes the extraction process complicated and the extraction cost high.
[0003] Rosa roxburghii is the fruit of the Rosa roxburghii plant of the Rosaceae family, which is widely distributed in Hubei, Sichuan, Guizhou, Yunnan, Guangxi and other places in my country. The fruit of Rosa roxburghii is oblate or conical, with a diameter of 2-4cm. The surface is yellow-brown and densely covered with thorns. The flesh is yellow-white; the seeds are borne on the raised receptacle at the base of the calyx tube. The folk use of Rosa roxburghii in my country has a long history. As early as more than 400 years ago, wild Rosa roxburghii fruits were used for winemaking. my country began to develop and protect wild Rosa roxburghii resources in the 1980s and promoted artificial cultivation. Artificial cultivation of Rosa roxburghii technology also provides a better source of raw materials for SOD extraction technology. Summary of the invention
[0004] In view of the shortcomings of the prior art, the purpose of the present invention is to provide a method for producing a liquid dosage form rich in superoxide dismutase activity by sugar soaking and thawing roxburghii fruit.
[0005] The second object of the present invention is to provide the product obtained by the extraction method.
[0006] The technical solution for achieving the above-mentioned purpose of the present invention is:
[0007] A method for producing a liquid dosage form rich in superoxide dismutase activity by sugar soaking after strong freezing and thawing of roxburghii fruit, comprising the following steps:
[0008] 1) Planting of roxburghii: Planting roxburghii artificially, picking the roxburghii fruits when they are 100% ripe (the picked fruits are called roxburghii fruits, or fresh roxburghii fruits).
[0009] 2) Use an old washing machine or a special burr removal device to remove the burrs of fresh roxburghii fruit. Rinse the burrs with water, dry the surface of the fresh fruit in the sun or shade, and then cut and remove the seeds;
[0010] 3) cutting and removing the seeds of the fresh roxburghii fruit and performing strong freezing, wherein the temperature of the strong freezing is 0 to -80°C;
[0011] 4) After thawing, soak the fruit in sugar at 1.8 times the weight of the fruit;
[0012] 5) After the reaction is completed, the juice is squeezed with a weight of more than 20 kg to obtain the juice.
[0013] Among them, the burrs of the fresh fruit of the roxburghii are removed by using a pulsator washing machine or a special burr removal device. The hair and thorns are washed clean with water, the surface moisture of the fresh fruit is dried in the sun or in the shade, and then it is cut and the seeds are removed. The purpose is to make the inside and outside areas of the fresh fruit of the roxburghii fully contact with the sugar water, so as to quickly and evenly breed the SOD activity.
[0014] According to experience, the SOD production of wild roxburghii is not good, so it is better to use artificially grown roxburghii fresh fruit. The soil in the mountainous areas of Guizhou and Guangxi where roxburghii is grown is thin, so compound fertilizers are not suitable, only organic fertilizers can be used. And add phosphorus fertilizer.
[0015] Preferably, the roxburghii roxburghii is artificially planted in mountainous areas, and phosphate fertilizer and organic fertilizer are applied during planting, and the weight ratio of phosphate fertilizer to organic fertilizer is 1:10-20.
[0016] Organic fertilizer can be decomposed chicken manure or commercially available organic fertilizer.
[0017] Wherein, in step 3), the strong freezing uses the freezer of a freezer or refrigerator, the strong freezing temperature is 0 to -30°C, and the strong freezing time is 2 to 3 days.
[0018] Furthermore, in step 3), the room temperature and product temperature are required to be 30-32°C, and the indoor floor is preferably kept wet for a long time. More preferably, the operation is carried out in an air-conditioned room.
[0019] A preferred technical solution of the present invention is that in step 3), when the temperature of the fresh fruit is raised to 30-32°C, the amount of sugar in each barrel or tank is 1.8 times the weight of the fresh fruit, the initial amount of sugar is 1.2 times the weight of the fruit, the sugar and fresh fruit are evenly mixed and poured into the barrel or tank, the fruit is turned over after 2 days, and then a small amount of 0.6 times the weight of the fruit is evenly sprinkled on the surface of the fresh fruit in the barrel or tank, the fruit is turned over once a day, and the sugar at the bottom of the barrel or tank is turned over on the surface of the fresh fruit, and this is done every day until 0.6 times the weight of the fruit The sugar is used up.
[0020] After sugar is added to the fresh fruit, the middle and late stage fresh fruit mixture floats on the surface of the barrel or jar. If the amount is small, the fresh fruit can be evenly soaked in the sugar water. If the barrel or jar is filled with a large amount, the fresh fruit floating on the surface cannot be soaked in the sugar water and cannot absorb oxygen evenly, which will affect the quality of the juice.
[0021] The reaction is completed when the reaction mixture stops bubbling.
[0022] The reaction of 100% mature fresh fruit is completed in 15-20 days (it reaches a saturated solution containing sugar when there is no bubbling).
[0023] Wherein, in step 4), filtering at a temperature of 35-42° C. and pressing with a weight of more than 20 kg are performed in an air-conditioned room.
[0024] The pomace after strong freezing and filtration is like being cooked in a pressure cooker. When pressed by a hydraulic press, the pomace and juice will splash out together. Therefore, double-layer barrels and heavy objects are used for pressing.
[0025] Another preferred technical solution of the present invention is that in step 5), a double-layer stainless steel barrel is used for squeezing and filtering, the height of the outer barrel 1 of the double-layer stainless steel barrel is 1.8 to 2.5 times the height of the inner barrel 2, and the inner diameter of the outer barrel is 1.5 to 3 cm larger than the outer diameter of the inner barrel; Φ1.5-2 mm sieve holes are drilled on the inner barrel wall and the bottom of the barrel.
[0026] Wherein, in step 5), the pomace of the fruit is put into the inner barrel by using a filter screen or a cloth bag, and then a heavy object weighing more than 20 kg is put into a plastic bag and placed on the pomace of the inner barrel to exert pressure.
[0027] The method of the invention prepares juice rich in SOD active enzyme.
[0028] Compared with the prior art, the present invention has the following beneficial effects:
[0029] The method for preparing superoxide dismutase from roxburghii by strong freezing proposed by the present invention can breed SOD in larger quantities. Compared with the breeding method without strong freezing, the advantages include:
[0030] 1. Use an old-fashioned washing machine or a special depilatory and thorn equipment to remove burrs, clean, cut and remove seeds (preferably dry the surface moisture), and then enter strong freezing. Because the fresh fruit of roxburghii encounters water, oxygen, light radiation SOD active enzyme, VC and other nutrients under natural conditions, the loss is very large. The inventor has done an experiment with bottled juice VC only under light, and the VC dropped by 25% after 12 months. After the fresh fruit is strongly frozen, the SOD active enzyme and its nutrients are not lost and the technical problem of not being able to keep fresh fruit for a long time is solved.
[0031] 2. The fresh frozen sea buckthorn fruit can be transported to other places for processing, which can reduce the production cost of product transportation by 200 times that of drinking water and 138 times that of wine.
[0032] 3. Fresh roxburghii fruits mature in August and September every year, and the temperature in Guizhou Province and nearby areas drops to between 15-20℃. The best temperature for sugar soaking is 30-32℃. The room temperature and product temperature for pressing require 35-42℃. Transporting frozen fruits to Guangxi, Guangdong, Hainan and other places, and setting up factories near water sources for processing can save a lot of production costs such as heating electricity and water transportation.
[0033] 4. Strongly frozen fresh fruits need to be thawed before they can be soaked in sugar. The lignin of the original fresh fruit is dense and rich in water. After being strongly frozen, the flesh has turned into ice cubes. After thawing, the lignin of the flesh changes from dense to loose. It is like a glass bottle filled with water and sealed tightly, and the bottle breaks after freezing. Sugar water quickly enters and breeds SOD active enzymes, and quickly flows out, unable to stay on the pomace, and the SOD active enzyme detection value of the resulting juice doubles.
[0034] The cost of SOD products produced by this method is less than 20 yuan / g active SOD. Compared with the method of extracting SOD from cow blood, the production cost is reduced by 3600 times.
[0035] 5. Biologists have confirmed that the active enzyme of liquid SOD is absorbed immediately after oral administration, and can penetrate the blood vessel wall in 5 seconds and enter the blood circulation for 136 hours. Cell tissues can remove free radicals through their unique affinity with free radicals, enhance the function of tissues and organs and human immunity, fight against the root causes of diseases and cancer, and make the body full of youthful vitality. This overcomes the absorption problem of ordinary capsules and tablets in the long 3-6 hours of gastrointestinal disintegration, where gastric acid and choline inactivate the SOD activity to less than 3%. BRIEF DESCRIPTION OF THE DRAWINGS
[0036] Figure 1 This is a schematic diagram of the double-layer stainless steel barrel structure.
[0037] In the figure, the parts and their numbers are: outer tube 1, inner barrel 2, cloth bag 3, and weight 4. DETAILED DESCRIPTION
[0038] The following examples are used to illustrate the present invention but are not intended to limit the scope of the present invention.
[0039] Unless otherwise specified, the raw materials and equipment used in the specification are commercially available; the technical means adopted are those known in the art.
[0040] The roxburghii in the embodiment is planted in the mountainous area of Dushan County, Guizhou Province.
[0041] Embodiment 1:
[0042] 1) Main raw materials: artificially planted fresh roxburghii fruit, the roxburghii is artificially planted roxburghii, organic fertilizer and a small amount of phosphate fertilizer are applied during planting, the organic fertilizer is decomposed chicken manure, and the phosphate fertilizer is superphosphate. Before planting, 3kg of organic fertilizer and 0.2kg of phosphate fertilizer are applied to each tree hole, and 0.2kg of phosphate fertilizer is applied to each tree in the second year of planting. After three years of planting, 0.5kg of organic fertilizer is applied to each tree every year.
[0043] 2) The fresh fruits of this species of sea buckthorn should be picked when they are 100% mature.
[0044] 3) Use an old washing machine or special equipment for burr removal to remove the burrs of fresh roxburghii fruit. Rinse the burrs with water, dry the surface of the fresh fruit in the sun or shade, and then cut and remove the seeds. (After the fresh fruit is cut and the seeds are removed, it is strictly forbidden to soak it in water to prevent a large amount of SOD activity from being lost.) The purpose is to make the inside and outside of the roxburghii fruit fully contact the sugar water to quickly and evenly breed SOD activity. Use the refrigerator freezer to freeze it at -18℃ for 2 days. After the fresh fruit is frozen and thawed, soak it in 1.8 times the weight of the fruit in sugar, and the juice obtained is 1.8 times the weight of the fruit.
[0045] 4) Use a stainless steel conical barrel (wide at the top and small at the bottom), put 10kg of seeded roxburghii fruit in each barrel, add 1.8 times the weight of the fruit with white sugar, and each barrel contains about 30kg of the mixture. (The juice is about 1.8 times the weight of the fruit). After adding sugar to the fresh fruit, the mixture of the middle and late fresh fruit floats on the barrel surface, and the small amount of fresh fruit is evenly soaked in the sugar water. Because the capacity of the barrel is large, a large number of fresh fruits floating on the surface cannot soak in the sugar water and cannot evenly absorb oxygen, which will affect the quality of the juice.
[0046] 5) When the room temperature reaches about 30-32℃, start adding sugar. Mix 1.2 times the weight of the fruit with the fresh fruit and pour it into the bucket. For the remaining sugar, turn the candies at the bottom of the bucket over the fruit surface every day and evenly cover them with a layer of sugar until all the remaining sugar is used up.
[0047] 6) 100% mature fresh fruit of the roxburghii roxburghii needs 15-20 days (SOD) active breeding reaction to complete, until there is no bubbling. The optimal temperature for active breeding is 30-32°C. Temperatures below 27-29°C or above 33-35°C will slow down the reaction or stop SOD active breeding. The reaction is carried out in an air-conditioned room.
[0048] 7) When the reaction is complete (no bubbles in the barrel), raise the room temperature and product temperature to 35-42℃, and use a coarse-mesh stainless steel filter to dry the juice. Then take the filtered pomace (product temperature 35-42℃) and add a weight of more than 20kg to squeeze it until no juice drips out of the pomace.
[0049] 8) Mix the filtered juice and the squeezed juice, and then finely filter them with a 60-80 mesh stainless steel filter at room temperature of 35-42°C. Under the above conditions, the juice is allowed to precipitate naturally for 1-2 days to obtain a product of SOD active juice of 1800g / kg of fresh roxburghii fruit, which is stored in a large barrel for later use.
[0050] 9) After strong freezing and thawing, the sugar-soaked pomace is like being cooked under high pressure. When it is pressed by a hydraulic press, the pomace and juice will be ejected together. Use double-layer stainless steel barrels (see structure for details) Figure 1 ), the height of the outer barrel 1 is about twice the height of the inner barrel 2, and the inner diameter of the outer barrel is 2 cm larger than the outer diameter of the inner barrel; the side wall and bottom of the inner barrel 2 are drilled with Φ1.5-2 mm sieve holes, and the pomace of the fruit is put into the inner barrel 2 with a cloth bag 3, and then a weight 4 of more than 20 kg is put in a plastic bag and placed on the pomace of the inner barrel to apply pressure, and the obtained juice is coarse filtered juice 1800 g / kg of fresh roxburghii fruit.
[0051] After the roxburghii fruit is strongly frozen and thawed, it is soaked in sugar to obtain SOD active juice which is 1.8 times of the fruit weight, of which about 80% is coarse filtered juice and about 20% is squeezed. Under the above conditions, the juice is naturally precipitated for 1-2 days to obtain the product SOD active juice 1800g / kg roxburghii fresh fruit.
[0052] The SOD active juice obtained in this example was sent to Guangzhou Analysis and Testing Center, China ) The test standard was GB / T-5009.171-2003 Method 1, and the SOD content was 3.36×10 5 U / 100mL.
[0053] Example 2
[0054] 1) Main raw materials: artificial planting of fresh roxburghii fruit, rock sugar or first-grade white sugar, sucrose, beet sugar, temperature, oxygen, humidity, etc. The planting process is the same as in Example 1.
[0055] 2) The fresh fruits of this species of sea buckthorn must be 100% ripe for picking.
[0056] 3) Use an old washing machine to remove the burrs of fresh roxburghii fruit. Rinse the burrs with water, dry the surface of the fruit in the sun or shade, then cut and remove the seeds. Use the refrigerator freezer to freeze at -18℃ for 2 days. After the fresh fruit is thawed, soak it in sugar twice the weight of the fruit, and the juice obtained is twice the weight of the fruit.
[0057] 4) Use porcelain jars to put about 12kg of seeded roxburghii fruit in each jar, add 2 times of sugar, and the mixture in each jar is about 30kg. The juice is about 2 times the weight of the fruit. After adding sugar to the fresh fruit, the mixture of the middle and late fresh fruit floats on the surface of the jar. The small amount of fresh fruit is evenly soaked in the sugar water. The large amount of fresh fruit floating on the surface cannot be soaked in the sugar water and cannot evenly absorb oxygen, which will affect the quality of the juice.
[0058] 5) The room and product temperature should be around 30-32℃ before adding sugar. Mix the sugar about 1.1 times the weight of the fruit with the fresh fruit and pour it evenly into the tank. For the remaining sugar, turn the candies that sink to the bottom of the tank over the fruit surface when turning the fruit every day, and evenly cover the fruit with a layer of sugar. Use up the remaining sugar.
[0059] 6) It takes 15-20 days for the (SOD) active growth reaction to be completed for 100% mature fresh roxburghii fruit.
[0060] 7) After the reaction is complete (no bubbling), raise the room temperature and product temperature to 35-42°C.
[0061] 8) using a double-layer stainless steel barrel (same as in Example 1), a weight of 20 kg was placed on the pomace of the inner barrel to apply pressure, and the resulting juice was coarsened and filtered to obtain 1800 g / kg of roxburghii fresh fruit.
[0062] The juice is allowed to settle naturally for 1-2 days to obtain the SOD active juice, which is then stored in large barrels for later use.
[0063] The SOD content of the SOD active juice obtained in this example was measured to be 2.73×10 5 U / 100mL. 2730U / ml×1800ml=4914000U, which is the active SOD produced by 1kg of roxburghii fruit. The cost of the product produced by this method is less than 20 yuan / g SOD.
[0064] Example 3
[0065] The roxburghii roxburghii is artificially planted, and organic-inorganic compound fertilizer is applied during the planting period.
[0066] The fresh fruits of sea buckthorn are picked when they are 80-90% ripe.
[0067] The operation is the same as in Example 1.
[0068] There are many fruits, and it takes more than 30 days and nights for the reaction to be complete.
[0069] Example 4
[0070] Plant sea buckthorn and pick the fresh fruits when they are 100% ripe.
[0071] The operation is basically the same as in Example 1, except that in step 5), sugar is added at room temperature of 28°C.
[0072] It takes 25 days for the SOD activity breeding reaction to be completed.
[0073] Example 5
[0074] This embodiment provides a beverage, which is the SOD active juice prepared in Example 1 packaged in a beverage bottle. The beverage bottle consists of a bottle body and a beverage bottle cap, and there is an area for containing SOD inside the beverage bottle cap. The top of the beverage bottle cap 4 and the bottom center of the area for containing SOD are sealed by a plastic film 2. The bottle body is filled with pure water, and the SOD active juice is placed in the area for containing SOD. When drinking, pierce the plastic film with a toothpick to mix the pure water and the SOD active juice.
[0075] Although the present invention has been described above through embodiments, those skilled in the art should understand that without departing from the spirit and substance of the present invention, all improvements and modifications made to the present invention should fall within the protection scope of the present invention.
Claims
1. A method for producing a liquid dosage form rich in superoxide dismutase activity by sugar soaking after strong freezing and thawing of roxburghii fruit, characterized in that: Includes operations: 1) Planting of roxburghii: Planting roxburghii artificially and picking the roxburghii fruits when they are 100% ripe. 2) Remove the burrs from the fresh fruit of the roxburghii roxburghii with an old-fashioned washing machine or a special burr removal device, rinse the burrs with water, dry the surface of the fresh fruit in the sun or in the shade, and then cut and remove the seeds; 3) subjecting the cut and seeded fresh roxburghii fruits to strong freezing at a temperature of 0 to -80°C; 4) After thawing, soak the fruit in sugar at 1.8 to 2.0 times the weight of the fruit; 5) After the reaction is completed, the juice is squeezed with a weight of more than 20 kg to obtain the juice.
2. The method according to claim 1, characterized in that: The roxburghii roxburghii is artificially planted in mountainous areas, and phosphate fertilizer and organic fertilizer are applied during the planting period, and the weight ratio of the phosphate fertilizer to the organic fertilizer is 1:10-20.
3. The method according to claim 1, characterized in that In step 3), the strong freezing is performed in a freezer or refrigerator at a temperature of 0 to -30°C and for 2 to 3 days.
4. The method according to claim 1, characterized in that: In step 3), the room temperature and product temperature are required to be 30-32°C, and the indoor floor is preferably kept wet for a long time.
5. The method according to claim 1, characterized in that In step 3), when the temperature of the fresh fruit is raised to 30-32°C, the amount of sugar added is 1.8 times the weight of the fresh fruit, and the initial amount of sugar is 1.2 times the weight of the fruit. The sugar and fresh fruit are evenly mixed and poured into a bucket or a vat; after 2 days, the fruit is turned over, and then a small amount of 0.6 times the weight of the fruit is evenly sprinkled on the surface of the fresh fruit in the bucket or the vat. The fruit is turned over once a day, and the sugar on the bottom of the bucket or the vat is turned over on the surface of the fresh fruit. This is done every day until 0.6 times the weight of the fruit is used up.
6. The method according to claim 1, characterized in that The reaction was complete when the reaction mixture stopped bubbling.
7. The method according to claim 1, characterized in that In step 5), filtering at a temperature of 35 to 42° C. and squeezing with a weight of more than 20 kg are performed in an air-conditioned room.
8. The method according to claim 1, characterized in that In step 5), a double-layer stainless steel barrel is used for squeezing and filtering, wherein the height of the outer barrel 1 of the double-layer stainless steel barrel is 1.8 to 2.5 times the height of the inner barrel 2, and the inner diameter of the outer barrel is 1.5 to 3 cm larger than the outer diameter of the inner barrel; and Φ1.5 to 2 mm sieve holes are drilled on the inner barrel wall and the bottom of the barrel.
9. The method according to claim 8, characterized in that In step 5), the pomace of the fruit is put into the inner barrel by using a filter screen or a cloth bag, and then a heavy object weighing more than 20 kg is put into a plastic bag and placed on the pomace of the inner barrel to apply pressure.
10. The juice rich in SOD active enzyme prepared by the method according to any one of claims 1 to 9.