A novel method for preparing corn superoxide dismutase multienzyme

Abstract

The invention discloses a method for preparing a novel corn superoxide dismutase multienzyme, and relates to the technical field of superoxide dismutase multienzyme preparation. The invention includes steps such as material preparation, germination, pulping, concentration, and drying. The invention selects corn from Liaoning, where the corn is produced, and the content of complex enzymes is high; a fully automatic seed cleaning, soaking, and germination device is used, and the processing capacity is large, 2t can be germinated each time, labor intensity is reduced, and the buds are evenly grown, and the content of complex enzymes is higher than that of ordinary germination methods, and large-scale industrial production can be achieved; a low-temperature vacuum concentrator is used to remove moisture at 22-25°C and concentrate, and at this temperature, the yield of complex enzyme activity is higher and the concentration multiple is higher. Experiments show that when the product of the present invention is stored at 45°C. The enzyme activity is not reduced within 90 days, and when the product of the present invention is stored at room temperature, the enzyme activity is not reduced within 48 months.

Description

Technical Field

[0001] The invention relates to the technical field of superoxide dismutase multienzyme production, in particular to a method for preparing a novel corn superoxide dismutase multienzyme. Background Art

[0002] The main effect of superoxide dismutase multi-enzyme on the human body is to effectively remove superoxide anions in the body - free radicals. Excessive free radicals stimulate cell proliferation changes and genetic mutations, which can cause a variety of diseases. Reduce cell damage or death in the body, thereby reducing the occurrence of diseases and delaying aging. There are two main methods for producing superoxide dismutase multi-enzymes. One is to extract from animal viscera, and the other is to extract from plants such as corn. There are many methods for preparing superoxide dismutase complex enzymes from corn, but there are problems with the use of chemical disinfectants such as bleaching powder and sodium hypochlorite in the preparation process. Since chemical disinfectants are easy to remain and pollute the environment during the production process, the quality of the prepared superoxide dismutase complex enzyme is also affected to a certain extent. At the same time, in the process of drying the product, the existing method uses freeze drying or spraying. The former has a higher cost and the latter has a larger loss of enzyme activity. Both are not suitable for large-scale production needs. The existing method still has technical problems such as low germination rate, low enzyme yield, and small production scale. Since corn is germinated in the box of the germination device, there is a problem of low ventilation. The corn growth environment is narrow, which easily causes bacterial film to form on the surface of the corn, resulting in corruption, resulting in a decrease in germination rate, affecting the quality of the obtained superoxide dismutase, and easily leaving residues, which is not easy to clean, increasing the labor intensity of workers.

[0003] Chinese patent, patent number CN201210160871.0, the name of the invention is industrial production method for preparing superoxide dismutase complex enzyme using corn as raw material and its germination chamber. This patent solves the above technical problems, but it still has the following technical problems: low germination rate and yield, low concentration efficiency, no clear indication of the specific type of complex enzyme produced, and no indication of the shelf life of the complex enzyme at high temperature. Summary of the invention

[0004] In order to solve the above problems, that is, to solve the problems raised by the above background technology, the present invention proposes a novel method for preparing corn superoxide dismutase multienzyme, and the specific technical scheme is as follows:

[0005] A novel method for preparing corn superoxide dismutase multienzyme, characterized in that the production method is as follows:

[0006] Preparation: prepare an appropriate amount of raw corn, remove impurities from the corn, and set aside;

[0007] Germination: Place the corn in a fully automatic seed cleaning, soaking and germination device, clean and soak for 6-15 hours, and germinate for 24-64 hours until the sprouts are 3-5 mm long;

[0008] Pulping: adding purified water to germinated corn in a ratio of 1:2-4, mixing and evenly crushing to form corn pulp, stirring the corn pulp evenly and filtering, collecting the filtrate, and adding 0.5%-1.5% of the weight of the filtrate to the filtrate;

[0009] Concentration and drying: placing the filtrate into a low-temperature vacuum concentrator and evaporating it at 22-25° C. for 8-15 hours to obtain a corn superoxide dismutase multi-enzyme liquid; mixing the corn superoxide dismutase multi-enzyme liquid and the active carrier evenly and placing the mixture into a drying oven and drying for 6-10 hours to obtain a corn superoxide dismutase multi-enzyme powder.

[0010] Further, the production method is as follows:

[0011] Preparation: prepare an appropriate amount of corn, remove impurities from the corn, and set aside;

[0012] Germination: Place the corn in a fully automatic seed cleaning, soaking and germination device, clean and soak for 10 hours, and germinate for 50 hours until the sprouts are 4 mm long;

[0013] Pulping: adding purified water to germinated corn in a ratio of 1:3, and evenly crushing the mixture into corn pulp, stirring the corn pulp evenly and filtering it, collecting the filtrate, and adding 1% antioxidant by weight of the filtrate to the filtrate;

[0014] Concentration and drying: placing the filtrate into a low-temperature vacuum concentrator and evaporating it at 22-25° C. for 12 hours to obtain a corn superoxide dismutase multi-enzyme liquid; mixing the corn superoxide dismutase multi-enzyme liquid and the active carrier evenly and placing the mixture into a drying oven and drying for 8 hours to obtain a corn superoxide dismutase multi-enzyme powder.

[0015] Furthermore, the corn is produced in Liaoning Province.

[0016] Furthermore, the germination process is specifically as follows: corn seeds are placed in a fully automatic seed cleaning, soaking and germination device, the temperature is set to 28-35°C for germination, compressed air is introduced into the device every 0.5 hour during the germination stage, water is sprayed into the device every 1 hour, and the water is drained after each spraying. After the germination is completed, the germinated corn is washed 3-5 times and set aside.

[0017] Furthermore, the antioxidant is one or both of apple polyphenols and vitamin C.

[0018] Furthermore, the active carrier is one or both of starch and polysaccharide.

[0019] Furthermore, the filtration step in the pulping process uses a plate and frame filter press.

[0020] Furthermore, a low-temperature vacuum concentration device with an evaporation temperature of 22-25° C. is used in the concentration step.

[0021] Furthermore, the superoxide dismutase multienzyme is specifically superoxide dismutase, glutathione peroxidase, and catalase.

[0022] The beneficial technical effect of the present invention is: corn from a special producing area of ​​Liaoning is selected, and the content of complex enzyme is high.

[0023] Patent CN201210160871.0 indicates that the germination chamber has a simple structure, large ventilation volume, and a high corn germination rate. Compared with patent CN201210160871.0, this application adopts a fully automatic seed cleaning, soaking, and germination device with a large processing capacity. Each can can germinate 2t each time, reducing labor intensity, and the buds are evenly grown. The complex enzyme content is higher than that of the ordinary germination method. And this equipment can complete the whole process of corn cleaning, soaking, germination, etc., with a high degree of automation, low energy consumption, small footprint, and large processing capacity, which is particularly suitable for the needs of industrialized mass production of corn superoxide dismutase multi-enzyme. During cleaning, the corn is cleaned by blasting and rolling, and floating impurities are automatically removed by overflow. During soaking, the appropriate soaking water temperature can be controlled. During germination, the compressed air ports at different locations can be used for staggered ventilation to maintain good ventilation between the germinating corns, making it difficult for microorganisms to breed. At the same time, the temperature of the spray water and the temperature of the germination tank can be controlled to maintain the appropriate germination temperature and moisture, so that the temperature and humidity conditions of the whole tank of corn are uniform, the germination rate is high, and the sprout length is uniform.

[0024] A low-temperature vacuum concentrator is used to remove moisture at 22-25°C for concentration. The evaporation efficiency is high and can reach 800L / h. At this temperature, the yield of the compound enzyme activity is higher and the concentration multiple is higher. The evaporation temperature of the traditional vacuum concentration equipment is above 50°C. Some temperature-sensitive enzymes and other biologically active substances are easily inactivated at this temperature, such as catalase and glutathione peroxidase. Even superoxide dismutase with good thermal stability has a low yield. The low-temperature vacuum concentration equipment used in this application can be concentrated at a temperature of 22-25°C, which can not only ensure that catalase and glutathione peroxidase are not inactivated, but also improve the yield of superoxide dismutase. At the same time, the concentration multiple of this equipment is relatively high, which can reach 40-60 times. Under the same extraction rate, the activity of the corn superoxide dismutase multi-enzyme liquid and powder per unit weight can be greatly improved. Finally, because this equipment adopts heat pump technology, heat source and cold source are provided, the purpose of improving efficiency, reducing energy consumption and saving space is achieved.

[0025] Experiments show that when the product of the present invention is stored at 45°C, the enzyme activity does not decrease within 90 days, and when the product of the present invention is stored at room temperature, the enzyme activity does not decrease within 48 months.

[0026] Compared with the patent number CN201210160871.0, whose invention name is "Industrial production method for preparing superoxide dismutase complex enzyme using corn as raw material and its germination chamber", the present application reduces the use of additives. During use, only apple polyphenols and vitamin C are added as antioxidants, and starch or polysaccharides are added as carriers. The rest are all taken from corn itself, reducing the use of phosphates, diatomaceous earth and seaweed polysaccharides in patent CN201210160871.0, thereby greatly reducing the cost of the product.

[0027] At the same time, due to the small number of by-products, there is only a certain amount of corn pulp residue by-product in the pulping step. This by-product can be used as a feed raw material, which can improve the meat flavor and reduce environmental pollution. DETAILED DESCRIPTION

[0028] The preferred embodiments of the present invention are described below. It should be understood by those skilled in the art that these embodiments are only used to explain the technical principles of the present invention and are not intended to limit the protection scope of the present invention.

[0029] Example 1

[0030] Prepare 2000kg corn, remove impurities from the corn, and set aside; place the corn in a fully automatic seed cleaning, soaking, and germination device, wash, soak for 6 hours, set the temperature to 28-35°C, germinate for 24 hours, introduce compressed air into the device every 0.5 hours during the germination stage, spray water into the device every 1 hour, drain the water after each spraying, germinate until the buds are 3mm long, wash the germinated corn 3-5 times, and set aside. Add purified water to the germinated corn at a ratio of 1:3, grind the mixture evenly into corn pulp, stir the corn pulp evenly, filter, collect the filtrate, and add 0.5% of apple polyphenols by weight of the filtrate to the filtrate. Place the filtrate in a low-temperature vacuum concentrator, evaporate at 22-25°C for 8 hours to obtain corn superoxide dismutase multi-enzyme liquid, mix the corn superoxide dismutase multi-enzyme liquid with starch evenly, place it in a drying oven, dry it for 6 hours, and obtain corn superoxide dismutase multi-enzyme powder.

[0031] Example 2

[0032] 2000kg corn is prepared, impurities in the corn are removed and set aside; the corn is placed in a fully automatic seed cleaning, soaking and germination device, washed and soaked for 10 hours, the temperature is set to 28-35°C, and the corn is germinated for 50 hours. During the germination stage, compressed air is introduced into the device every 0.5 hours, water is sprayed into the device every 1 hour, and the water is drained after each spraying. The germination is carried out until the buds are 4mm long, and the germinated corn is washed 3-5 times and set aside. Purified water is added to the germinated corn in a ratio of 1:3, the mixture is evenly crushed into corn pulp, the corn pulp is stirred evenly and then filtered, the filtrate is collected, and 1% of the weight of the filtrate vitamin C is added to the filtrate. The filtrate is placed in a low-temperature vacuum concentrator, evaporated at 22-25°C for 12 hours, and a corn superoxide dismutase multi-enzyme liquid is obtained. The corn superoxide dismutase multi-enzyme liquid is mixed evenly with polysaccharide and then placed in a drying oven, dried for 8 hours, and a corn superoxide dismutase multi-enzyme powder is obtained.

[0033] Example 3

[0034] 2000kg corn is prepared, impurities in the corn are removed and set aside; the corn is placed in a fully automatic seed cleaning, soaking and germination device, washed and soaked for 15 hours, the temperature is set to 28-35°C, and the germination is carried out for 64 hours. During the germination stage, compressed air is introduced into the device every 0.5 hours, water is sprayed into the device every 1 hour, and the water is drained after each spraying. The germination is carried out until the buds are 5mm long, and the corn after germination is washed 3-5 times and set aside. Purified water is added to the germinated corn in a ratio of 1:3, the mixture is evenly crushed into corn pulp, the corn pulp is stirred evenly and then filtered, the filtrate is collected, and 1.5% of apple polyphenols by weight of the filtrate is added to the filtrate. The filtrate is placed in a low-temperature vacuum concentrator, evaporated at 22-25°C for 15 hours, and a corn superoxide dismutase multi-enzyme liquid is obtained. The corn superoxide dismutase multi-enzyme liquid is mixed evenly with starch and then placed in a drying oven, dried for 10 hours, and a corn superoxide dismutase multi-enzyme powder is obtained.

[0035] During the germination process, corn seeds produce a large amount of antioxidant enzyme substances, among which superoxide dismutase (SOD), glutathione peroxidase (GSH-px), and catalase (CAT) are the three most important enzymes. They work together to complete the entire process of removing oxygen free radicals, which is complete and efficient. Corn superoxide dismutase multienzyme is rich in these three enzymes.

[0036]

[0037]

[0038]

[0039] Superoxide dismutase (SOD) converts superoxide anion oxygen free radical It is converted into hydrogen peroxide (H2O2) and oxygen (O2). However, hydrogen peroxide (H2O2) still has strong oxidative activity and is prone to produce free radicals. Through catalase (CAT) and glutathione peroxidase (GSH-px), hydrogen peroxide (H2O2) is converted into water without oxidative activity, completing the entire process of removing free radicals.

[0040] The three enzymes work together to complete the whole process of removing free radicals and anti-oxidation. Only one or two of them can not complete the whole process, which affects the anti-oxidation effect.

[0041] Catalase and glutathione peroxidase are sensitive to temperature and easily inactivated in a low concentration liquid state. Therefore, these two enzymes are not described in other inventions of corn superoxide dismutase multienzymes, because the evaporation temperature of the reduced pressure concentration equipment they use is generally above 50°C. The low-temperature vacuum concentration equipment we use can be highly concentrated at 22-25°C, which can protect the activity of these two enzymes in the heat-sensitive stage to the greatest extent. When the two enzymes are concentrated more than 30 times and under the protection of antioxidants, their activity can have good thermal stability, which is convenient for the subsequent drying process.

[0042] The corn superoxide dismutase multi-enzyme powder obtained in Examples 1-3 was compared with the superoxide dismutase complex enzyme obtained in Patent CN201210160871.0. The test results are as follows:

[0043]

[0044] The experimental results show that the shelf life and enzyme activity of the superoxide dismutase multi-enzyme powder obtained in the present invention are better than those of the superoxide dismutase complex enzyme preparation obtained in patent CN201210160871.0. And under the same conditions, Example 3 has the best effect.

[0045] Although the present invention has been described with reference to preferred embodiments, various modifications may be made thereto and parts thereof may be replaced by equivalents without departing from the scope of the present invention. In particular, the various technical features mentioned in the various embodiments may be combined in any manner as long as there is no structural conflict. The present invention is not limited to the specific embodiments disclosed herein, but includes all technical solutions falling within the scope of the claims.

[0046] So far, the preferred embodiments shown describe the technical solutions of the present invention, but it is easy for those skilled in the art to understand that the protection scope of the present invention is obviously not limited to these specific embodiments. Without departing from the principle of the present invention, those skilled in the art can make equivalent changes or substitutions to the relevant technical features, and the technical solutions after these changes or substitutions will fall within the protection scope of the present invention.

Claims

1. A method for preparing corn superoxide dismutase multienzyme, characterized in that: The preparation method is as follows: Preparation: prepare an appropriate amount of raw corn, remove impurities from the corn, and set aside; Germination: placing corn in a fully automatic seed cleaning, soaking and germination device, cleaning, soaking for 6-15 hours, germinating for 24-64 hours, until the buds are 3-5 mm long; wherein the germination process is further as follows: placing corn seeds in a fully automatic seed cleaning, soaking and germination device, setting the temperature to 28-35° C., germinating, introducing compressed air into the device every 0.5 hour during the germination stage, spraying water into the device every 1 hour, draining the water after each spraying, and after the germination is completed, washing the germinated corn 3-5 times for standby use; Pulping: adding purified water to germinated corn in a ratio of 1:2-4, mixing and evenly crushing to form corn pulp, stirring the corn pulp evenly and filtering, collecting the filtrate, and adding 0.5%-1.5% of the weight of the filtrate antioxidant to the filtrate; Concentration and drying: placing the filtrate into a low-temperature vacuum concentrator and evaporating it at 22-25° C. for 8-15 hours to obtain a corn superoxide dismutase multi-enzyme liquid; mixing the corn superoxide dismutase multi-enzyme liquid and the active carrier evenly and placing the mixture into a drying oven for drying for 6-10 hours to obtain a corn superoxide dismutase multi-enzyme powder; The superoxide dismutase multienzyme is specifically superoxide dismutase, glutathione peroxidase, and catalase.

2. The method for preparing a corn superoxide dismutase multienzyme according to claim 1, characterized in that: The preparation method is as follows: Preparation: prepare an appropriate amount of corn, remove impurities from the corn, and set aside; Germination: Place the corn in a fully automatic seed cleaning, soaking and germination device, clean and soak for 10 hours, and germinate for 50 hours until the sprouts are 4 mm long; Pulping: adding purified water to germinated corn in a ratio of 1:3, and evenly crushing the mixture into corn pulp, stirring the corn pulp evenly and filtering it, collecting the filtrate, and adding 1% antioxidant by weight of the filtrate to the filtrate; Concentration and drying: placing the filtrate into a low-temperature vacuum concentrator and evaporating it at 22-25° C. for 12 hours to obtain a corn superoxide dismutase multi-enzyme liquid; mixing the corn superoxide dismutase multi-enzyme liquid and the active carrier evenly and placing the mixture into a drying oven and drying for 8 hours to obtain a corn superoxide dismutase multi-enzyme powder.

3. A method for preparing a corn superoxide dismutase multienzyme according to claim 1 or 2, characterized in that: The corn is produced in Liaoning Province.

4. A method for preparing a corn superoxide dismutase multienzyme according to claim 1 or 2, characterized in that: The antioxidant is one or both of apple polyphenols and vitamin C.

5. The method for preparing a corn superoxide dismutase multienzyme according to claim 1 or 2, characterized in that: The active carrier is one or both of starch and polysaccharide.

6. A method for preparing a corn superoxide dismutase multienzyme according to claim 1 or 2, characterized in that: The filtration step in the pulping process adopts a plate and frame filter press.

7. The method for preparing a corn superoxide dismutase multienzyme according to claim 6, characterized in that: The concentration step uses a low-temperature vacuum concentration device with an evaporation temperature of 22-25°C.

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