CN112914109B

CN112914109B - Preparation method and application of peptide nano-selenium

Info

Publication number: CN112914109B
Application number: CN202110160008.4A
Authority: CN
Inventors: 陈玉祥; 黄湛明; 何继军
Original assignee: Individual
Current assignee: Individual
Priority date: 2021-02-05
Filing date: 2021-02-05
Publication date: 2022-12-20
Anticipated expiration: 2041-02-05
Also published as: CN112914109A

Abstract

The invention discloses a preparation method and application of peptide nano-selenium, belonging to the technical field of deep processing of foods. A process for preparing nano-Se peptide includes reaction between Se ore and organic acid under electromagnetic catalysis, and culturing plant in the culture liquid. According to the invention, the reaction of the selenium ore and the organic acid is electromagnetically catalyzed, so that selenium in the selenium ore can be converted into free selenium ions and further converted into selenate ions, and therefore, the utilization rate of plants on selenium in the ore can be improved and further converted into peptide nano selenium; the process does not need to add a chemical catalyst, and accords with the concept of green chemistry; the culture solution prepared from selenium ore is used for selenium-rich culture of plants, the culture period is short, and the yield of peptide nano selenium is improved.

Description

Preparation method and application of peptide nano-selenium

Technical Field

The invention belongs to the field of deep processing of foods, and particularly relates to a preparation method and application of peptide nano-selenium.

Background

The human body is composed of more than 60 elements. According to the different contents of elements in human body, it can be divided into two categories of macroelements and microelements. At present, there are 18 kinds of essential trace elements that have been confirmed to be related to human health and life, i.e., iron, copper, zinc, cobalt, manganese, chromium, selenium, iodine, nickel, fluorine, molybdenum, vanadium, tin, silicon, strontium, boron, rubidium, arsenic, etc. Each of these trace elements has its own specific physiological function. Although they are present in very small amounts in humans, they are essential for maintaining some crucial metabolism in humans. Once these essential trace elements are absent, the human body can become ill and even life-threatening.

Selenium is used as a trace element and has multiple effects on human health. The method comprises the following steps:

and (3) regulating immunity: studies have shown that selenium can affect the immune system (including cellular, humoral, and non-specific immunity); for tumor patients, the decline of immune function and the deterioration of disease are vicious cycles, and the radiation therapy can also cause damage to the immune system; therefore, clinically, a proper amount of selenium is often supplemented to the tumor patients to improve the immunity of the organism and prevent and resist cancers.

Antioxidation: the main existing forms of selenium in human body are selenase and selenoprotein, wherein the selenium in glutathione peroxidase accounts for about 1/3 of the total selenium in human body. Glutathione peroxidase reduces peroxides harmful to the human body, thereby assisting in scavenging free radicals and protecting biofilms from oxidative damage by peroxides.

Inhibiting tumor growth: research finds that selenium can promote the growth of anti-tumor angiogenesis inhibiting factors and form an anti-tumor angiogenesis environment, so that the formation and development of tumor angiogenesis are inhibited, and a nutrition supply channel of tumor cells is cut off; under the same condition, selenium has certain protection effect on normal cells, and the phenomenon is more prominent under the condition of oxygen deficiency. Similar effects were also found in laboratory cell cultures. Relatively speaking, the probability of suffering from tumor is higher for the selenium-deficient people; the patients with selenium-deficient tumors are more likely to suffer from adverse phenomena affecting life cycle, such as lesion metastasis, multiple tumors, tumor deterioration, and the like.

Protecting the liver: the effect of selenium in liver protection is proved by a plurality of researchers, researchers find that the incidence rate of hepatitis patients can be reduced by 56% by selenium supplement, the recovery of the patients is promoted, and the researchers find that transaminase of the alcoholic liver patients can be recovered to be normal by selenium supplement treatment, the serum cholesterol content can be reduced, the low-density lipoprotein content can be reduced, the high-density lipoprotein content can be increased, and the effect of treating the fatty liver can be achieved. In the treatment of liver diseases, selenium and liver treatment medicines with toxic and side effects have good synergistic effect when taken together. The action mechanism of selenium for protecting the liver is as follows: the antioxidant effect of glutathione peroxidase and the promoting effect of selenium on human body immunity.

Antiviral ability: statistical data indicate that when a human body is short of selenium, the risk of infecting highly pathogenic viral diseases is obviously increased. Such as coxsackie virus, common cold virus, aids virus, ebola virus, smallpox virus, hepatitis virus, and even the novel coronavirus (COVID-19), are associated with selenium levels in patients.

In addition, selenium deficiency in humans can lead to cardiomyopathy, coronary heart disease, hypertension, diabetes, gastric disorders, decreased brain function (affecting the rate of metabolism of the brain 'neurotransmitter'), decreased vision, decreased thyroid function, and reduced male sexual function.

Therefore, it is very important to supplement selenium to the human body in a targeted manner, but not all of the selenium can be absorbed by the human body. Some selenium species can be absorbed by the human body, but have high toxicity. In contrast, organic selenium has a higher utilization than inorganic selenium, while having a lower toxicity than inorganic selenium. The method of natural selenium supplement is more beneficial to health than the method of taking inorganic selenium. However, in common edible crops, the content of organic selenium is low.

Disclosure of Invention

The present invention is directed to solving at least one of the problems of the prior art described above. Therefore, the invention provides a preparation method of peptide nano-selenium, which can enable plants to efficiently utilize selenium in ores and improve the content of the peptide nano-selenium in the plants.

The invention also provides the application of the peptide nano-selenium in the field of food and medicine preparation.

According to one aspect of the invention, the preparation method of the peptide nano-selenium is provided, which comprises the steps of reacting selenium ore with organic acid under electromagnetic catalysis, and taking a diluent of a system after reaction as a culture solution to culture plants.

According to a preferred embodiment of the invention, at least the following advantages are achieved:

(1) The selenium ore reacts with the organic acid under the electromagnetic catalysis, so that selenium in the selenium ore can be converted into free selenium ions and further converted into selenate ions, and therefore the utilization rate of plants to the selenium can be improved.

(2) The invention uses the diluent of the culture solution prepared by selenium ore to culture the selenium-rich of the plants, the culture period is short, and the yield of the peptide nano-selenium is improved.

(3) The preparation method of the peptide nano-selenium provided by the invention does not need to utilize a chemical catalyst, and is more in line with the concept of green chemistry.

(4) The content of the peptide nano selenium in the plant obtained by the method provided by the invention is as high as 340mg/kg.

In some embodiments of the present invention, the method for preparing peptide nano-selenium comprises the following steps:

s1, crushing the selenium ore to obtain selenium ore powder;

s2, adding the organic acid into the selenium ore powder, and performing electromagnetic catalytic reaction to obtain a selenium concentrated solution;

s3, taking a diluent of the selenium concentrated solution as a culture solution, and carrying out selenium-rich culture on the plants;

and S4, extracting nutrient substances of the plants obtained in the step S3, and carrying out enzymolysis to obtain the peptide nano selenium.

In some embodiments of the present invention, in step S1, the selenium ore powder has a particle size of 60 to 80 mesh.

The purpose of crushing the selenium ore is to increase the specific surface area of the selenium ore and improve the efficiency of the reaction in the step S2.

In some embodiments of the present invention, in step S2, the organic acid is at least one of carboxylic acid, sulfonic acid, sulfinic acid, and thiocarboxylic acid.

In some embodiments of the invention, the carboxylic acid is at least one of formic acid, acetic acid, octanoic acid, halogenated acids, hydroxy acids, keto acids, and amino acids.

In some embodiments of the invention, in step S2, the organic acid is purchased commercially without the addition of water-diluted concentrated acid.

In the step S2, some organic acids can participate in animal and plant metabolism and have obvious biological activity, so that the organic acids are used for preparing the culture solution and have no harm to the growth of plants and the individuals eating the plants.

In some embodiments of the present invention, in step S2, the mass ratio of the organic acid to the selenium ore powder is (2 to 50): 1.

the organic acid reacts with selenium in the selenium ore to generate selenate on one hand; another aspect is the effect of pH adjustment.

In some embodiments of the invention, step S2 is an electromagnetic catalytic reaction, the electromagnetic frequency being 0.3GHz to 300GHz.

In some embodiments of the invention, step S2 is an electromagnetic catalytic reaction, electromagnetic wavelength 10 ^-3 m～1m。

The electromagnetic catalytic reaction has the following functions: the high-frequency oscillation of the electromagnetic wave can accelerate the electronic transition of selenium in the selenium ore, and further convert the selenium in the selenium ore into ionic water-soluble selenium.

The organic acid has the following functions: reacting with ionic selenium generated by electromagnetic catalysis to generate stable and controllable selenate ions.

Under the weak acid environment, the plant utilization rate of the selenate ions is high, and the plants can utilize the selenate ions more quickly.

In some embodiments of the present invention, in step S2, the selenium concentration in the selenium concentrated solution is 20% to 45% by mass.

In some embodiments of the invention, in step S2, the selenium concentrate has a pH of 4.5 to 6.8.

In some embodiments of the present invention, in step S3, the dilution ratio of the diluted solution is 10 to 20 times, that is, each milliliter of selenium concentrated solution is diluted to 10 to 20ml with distilled water.

In some embodiments of the invention, in step S3, the pH in the culture solution is 4.5 to 6.8.

In some embodiments of the present invention, in step S3, the plant is at least one of common crops such as peanut, malt, corn, barley, soybean, and the like, vegetables such as cabbage, garlic, and the like, and fruit plants.

In some embodiments of the invention, in step S3, the selenium-enriched culture is performed for a period of 1 to 3 weeks.

In some embodiments of the invention, in step S3, the selenium-enriched culture is performed at a temperature of 20 ℃ to 40 ℃ under an illumination of 5 × 10 ⁴ ～7×10 ⁴ lx。

In some embodiments of the present invention, the plant or fruit cultured in step S3 may be sold without performing the step S4 according to the needs of the consumer.

In some embodiments of the invention, in step S4, the extracting comprises: cleaning and juicing the plants or fruits obtained in the step S3, adjusting the obtained juice to 40-60 ℃, adjusting the pH to 4.5-6.8, adding the complex enzyme, and standing for 12 hours under the condition of heat preservation; cooling to normal temperature, filtering to obtain filtrate, and obtaining the peptide nano selenium.

In some embodiments of the present invention, in step S4, the extraction subject may be the whole plant of the plant obtained in step S3 or a fruit part of the plant.

In some embodiments of the present invention, in step S4, the enzyme is at least one of thiol protease and metalloproteinase.

The peptide nano-selenium is applied to the field of food and medicine preparation.

In some embodiments of the present invention, the peptide nano-selenium can be directly eaten or can be combined with other ingredients to prepare foods or medicines for supplementing selenium to animals or human bodies.

Detailed Description

The idea of the invention and the resulting technical effects will be clearly and completely described below in connection with the embodiments, so that the objects, features and effects of the invention can be fully understood. It is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments, and other embodiments obtained by those skilled in the art without inventive efforts are within the protection scope of the present invention based on the embodiments of the present invention.

Example 1

The preparation method of the peptide nano-selenium comprises the following specific steps:

s1, crushing selenium ore powder to 80 meshes to obtain selenium ore powder;

s2, adding acetic acid into the selenium ore powder, wherein the mass of the acetic acid is 2 times that of the selenium ore powder;

s3, performing electromagnetic catalytic reaction on the system obtained in the step S2, wherein the frequency of the electromagnetic catalysis is 0.3GHz, the wavelength is 1m, and obtaining selenium concentrated solution with pH =6.8 after the reaction is completed;

s4, diluting the selenium concentrated solution obtained in the step S3 by 20 times, taking the diluted solution as a culture solution, and performing selenium-rich culture on the peanuts for 3 weeks, wherein before the selenium-rich culture, the peanut seeds are soaked in water for 24 hours at normal temperature;

and S5, extracting nutrient substances of the peanuts (100 whole plants) obtained in the step S4, and carrying out enzymolysis to obtain the peptide nano selenium.

Comparative example 1

The difference between the preparation of the peptide nano-selenium and the example 1 is that the specific process is as follows:

(1) Step S3 is not included, namely, the system obtained in step S2 is diluted by 20 times to be used as a culture solution, and the peanut is cultured for 3 weeks.

Comparative example 2

The comparative example prepares titanium nano-selenium, and the specific process is as follows:

s1, preparing Hoagland nutrient solution, wherein a specific formula is shown in a table 1;

s2, using the nutrient solution obtained in the step S1 as a culture solution, carrying out selenium-rich culture on the peanuts for 3 weeks, and soaking the peanut seeds in water for 24 hours at normal temperature before the selenium-rich culture;

and S3, extracting nutrient substances of the peanuts (100 whole plants) obtained in the step S4, and performing enzymolysis to obtain the peptide nano-selenium.

Table 1 hodgkin's nutrient solution formula.

The preparation method of the iron salt solution comprises the following steps: adding 2.78g of ferrous sulfate heptahydrate and 3.73g of disodium ethylene diamine tetraacetate into 500ml of distilled water;

the microelement solution is mixed water solution, and the solute concentration in the solution is 0.83mg/L potassium iodide, 6.2mg/L boric acid, 22.3mg/L manganese sulfate, 8.6mg/L zinc sulfate, 0.25mg/L sodium molybdate, 0.025mg/L copper sulfate and 0.025mg/L cobalt chloride.

Comparative example 3

The comparative example prepares the titanium nano-selenium, and the specific process comprises the following steps:

s1, crushing selenium ore to 80 meshes;

s2, uniformly mixing the selenium ore powder obtained in the step S1 with peat soil 1 in a mass ratio of;

s3, planting the peanut seeds soaked in water for 24 hours at normal temperature into the mixed system obtained in the step S2, watering with purified water, and planting for 3 weeks;

and S4, extracting nutrient substances of the peanuts (100 whole plants) obtained in the step S3, and carrying out enzymolysis to obtain the peptide nano-selenium.

Comparative example 4

s1, preparing a culture solution with the mass concentration of 1% and the pH value of 6.8 by using sodium selenite (CAS: 10102-18-8) and acetic acid as raw materials;

s2, taking the selenium concentrated solution obtained in the step S1 as a culture solution, and performing selenium-rich culture on the peanuts for 3 weeks, wherein before the selenium-rich culture, the peanut seeds are soaked in water for 24 hours at normal temperature;

and S3, extracting nutrient substances of the peanuts (100 whole plants) obtained in the step S2, and carrying out enzymolysis to obtain the peptide nano-selenium.

Test examples

The experimental example tests the content of the peptide nano selenium in the peanut plants obtained in the examples and the comparative examples, and the cost for preparing the peptide nano selenium in the peanut plants obtained in the examples 1 and the comparative examples 4.

Wherein, the calculation of the content of the peptide nano selenium refers to the formula: content = mass of peptide nano selenium/mass of plant material used; the calculation results are shown in table 2.

In the examples and comparative examples, the testing of peptide nanoselenium was carried out with reference to the national standard document GB 5009.93-2017.

Table 2 content of peptide nanoseles in examples and comparative examples.

Description of the preferred embodiment	Example 1	Comparative example 1	Comparative example 2	Comparative example 3	Comparative example 4
						Content (wt.)	340mg/kg	10mg/kg	2mg/kg	11mg/kg	300mg/kg

The cost accounting is shown in table 3.

Table 3 cost accounting for example 1 and comparative example 4.

The calculation method of the cost reduction comprises the following steps: the product of the unit price and the dosage of the material in the example 4 is subtracted by the product of the unit price and the dosage of the material in the example 1, and the difference is divided by the product of the unit price and the dosage of the material in the example 4.

In conclusion, the preparation method of the peptide nano-selenium provided by the application has high utilization rate of selenium in ores, and the content of the peptide nano-selenium in a single plant is as high as 340mg/kg; compared with the selenium-rich culture with sodium selenate culture nutrient solution, the cost is reduced by about 91 percent.

Application example

In the application example, the peptide nano selenium obtained in the examples and the comparative examples is applied to mice with malignant ascites to observe the treatment and recovery effects of the peptide nano selenium on the mice.

The modeling method comprises the following steps: mice were injected intraperitoneally with 0.1mL of EAC cells (cell number 1X 10) ⁶ One). And judging the modeling effect after six days.

Dividing the modeled mice with malignant ascites into 3 groups of 10 mice, wherein the control group is intraperitoneally injected with normal saline (9 mg/kg) every day; the standard group was given daily drug therapy (carboplatin, 9 mg/kg) to mice, and example 1 was injected daily with physiological saline (1 mg/mL,9 mg/kg) containing the corresponding peptide nanoselenium.

Mice were dissected 14 days after administration and tested for recovery of malignant ascites in mice in example 1, the standard group and the control group. The test results are shown in table 4.

Table 4 therapeutic effect of peptide nanoselenium on malignant ascites in mice.

Group of	Example 1	Control group	Standard group
				Ascites volume ml	2.1	22	1.6

The results show that the peptide nano-selenium prepared by the invention has excellent biological activity and obvious effect on the treatment of malignant ascites of mice. The embodiments of the present invention have been described in detail, but the present invention is not limited to the embodiments, and various changes can be made without departing from the gist of the present invention within the knowledge of those skilled in the art. Furthermore, the embodiments of the present invention and the features of the embodiments may be combined with each other without conflict.

Claims

1. A preparation method of peptide nano-selenium is characterized by comprising the following steps:

s1, crushing selenium ore to obtain selenium ore powder;

s2, adding organic acid into the selenium ore powder, and performing electromagnetic catalytic reaction to obtain a selenium concentrated solution;

s3, taking a diluent of the selenium concentrated solution as a culture solution, and performing selenium-rich culture on the plants;

s4, extracting nutrient substances of the plants obtained in the step S3, and performing enzymolysis to obtain the peptide nano-selenium;

in step S2, the organic acid is at least one of carboxylic acid, sulfonic acid, sulfinic acid and thiocarboxylic acid;

the carboxylic acid is at least one of formic acid and acetic acid.

2. The preparation method according to claim 1, wherein in step S1, the selenium ore powder has a particle size of 60 to 80 mesh.

3. The preparation method according to claim 1, wherein in step S2, the mass ratio of the organic acid to the selenium ore powder is (2 to 50): 1.

4. the preparation method according to claim 1, wherein in the step S2, the frequency of the electromagnetic catalytic reaction is 0.3GHz to 300GHz.

5. The preparation method of claim 1, wherein in the step S2, the mass concentration of selenium in the selenium concentrated solution is 20% -45%.

6. The method according to claim 1, wherein the pH of the culture medium in step S3 is 4.5 to 6.8.

7. The method according to claim 1, wherein in step S4, the enzymatic hydrolysis is carried out using at least one enzyme selected from thiol protease and metalloproteinase.