Preparation method of selenium-rich soybean enzymatic hydrolysate with blood lipid and blood sugar functions and application thereof

Abstract

The present application belongs to the field of health food, and particularly relates to a preparation method of selenium-rich soybean enzymatic hydrolysate with blood lipid and blood sugar lowering function and application thereof. The present application reasonably designs the components and proportion of the selenium-rich nutrient solution, uses ultrasonic treatment to promote soybean absorption and conversion of inorganic selenium into organic selenium in cooperation with vitamins, simultaneously uses multi-enzyme cooperation, combines specific ultrasonic and enzymatic hydrolysis, significantly improves the selenium-rich capacity of soybean, shortens the production time of selenium-rich soybean and solves other problems, and the prepared selenium-rich soybean enzymatic hydrolysate has blood lipid and blood sugar lowering function. Compared with non-selenium-rich soybean enzymatic hydrolysate and conventional diet, the selenium-rich soybean enzymatic hydrolysate can simultaneously improve the blood lipid and blood sugar conditions of patients with hyperlipidemia and hyperglycemia. In addition, the selenium-rich soybean enzymatic hydrolysate produced by the method of the present application has high total selenium and organic selenium content, short production cycle, blood lipid and blood sugar lowering activity, is safe, green and environmentally friendly, and has wide application prospect.

Description

Technical Field

[0001] This invention relates to a method for preparing selenium-enriched soybean enzymatic hydrolysate with lipid-lowering and blood sugar-lowering functions and its application, belonging to the pharmaceutical field. Background Technology

[0002] With the improvement of people's living standards, the acceleration of the pace of life, the increase in life pressure, the reduction of physical labor and exercise, and the deterioration of the work and environment, the body's metabolic capacity is prone to decline and disorder, the immunity is weakened, and the proportion of sub-healthy people is constantly rising. According to the latest data, high blood sugar is the leading cause of diabetes, so controlling blood lipid and blood sugar levels has become an urgent task. At present, patients with high blood lipid and high blood sugar mainly rely on statins and biguanides to control their blood lipid and blood sugar levels. However, long-term use of statins can easily damage muscle and liver tissue, while long-term use of biguanides may lead to hypoglycemia, lactic acidosis, and diabetic ketoacidosis. In addition, patients with high blood lipid often also have high blood sugar levels. Therefore, developing safe and effective products that can simultaneously lower blood lipids and blood sugar has important economic and social significance.

[0003] Selenium is an essential trace element for the human body. Insufficient selenium intake is closely related to the occurrence of more than forty diseases that seriously threaten human health (such as cancer, diabetes, cataracts, etc.) (Gao Xianli, Ye Chao, Ma Haile, et al. Research advances in preparation, stability, application, and possible risks of nanoselenium: Focus on food and food-related fields[J]. Journal of Agricultural and Food Chemistry, 2023, 71(23), 8731-8745.). Currently, selenium in selenium supplements mainly exists in the form of inorganic selenium (sodium selenite, sodium selenate), organic selenium (selenium-enriched crops and foods), and nanoselenium, but all of these have certain problems in practical applications. While inorganic selenium is inexpensive, it is highly toxic and can easily cause safety problems if not properly controlled. Organic selenium, while safe, has a long production cycle and high cost, making it economically unaffordable for most consumers. Furthermore, selenium-enriched crops and foods often have low organic selenium content, failing to significantly increase selenium intake (Zhang Maoming, Yang Xiaohe, Yao Liangliang, et al. Effects of bioactive selenium nutrient solution on soybean agronomic morphology, yield, and selenium content [J]. Heilongjiang Agricultural Sciences, 2021, (1), 34-36). In addition, selenium-enriched crops often have high levels of other heavy metals. Although nano-selenium has high bioavailability and relatively low cost, it cannot be legally applied to food selenium fortification. Therefore, how to economically and quickly convert inorganic selenium into organic selenium and develop selenium-enriched foods with high organic selenium content is of great significance. Summary of the Invention

[0004] To address the shortcomings of existing technologies, the purpose of this invention is to develop a selenium-enriched soybean enzymatic hydrolysate with lipid-lowering and blood sugar-lowering functions. Using inorganic selenium (sodium selenite, sodium selenate, nano selenium (1-1000μm)) and soybeans as raw materials, and supplemented by ultrasonic and enzymatic hydrolysis technologies, a selenium-enriched soybean enzymatic hydrolysate was developed, which has good lipid-lowering and blood sugar-lowering effects.

[0005] To achieve the above technical objectives, the present invention adopts the following technical solution;

[0006] A method for preparing selenium-enriched soybean enzymatic hydrolysate with lipid-lowering and blood glucose-lowering functions, comprising the following steps:

[0007] (1) Wash the soybeans with clean water and set aside;

[0008] (2) Soak soybeans in a selenium-enriched nutrient solution for 10-30 hours, applying ultrasound treatment during the soaking period (to promote the absorption and conversion of selenium and other nutrients) until the soaking is complete. After soaking, drain the water to obtain ultrasonically soaked soybeans. The selenium-enriched nutrient solution consists of selenium, vitamin C, vitamin B1, vitamin B7, and water, wherein the selenium content is 12-60 mg / L, and the selenium is derived from sodium selenite, sodium selenate, and nano-selenium. The contents of vitamin C, vitamin B1, and vitamin B7 are 500-5000 mg / L, 2-20 mg / L, and 10-100 μg / L, respectively. The conditions for the ultrasound treatment are: ultrasound frequency 20-65 kHz, and ultrasound intensity density 0.1-1.5 W / cm³. 2 Ultrasound time: 3-30 minutes;

[0009] (3) Soybeans soaked in ultrasound in step (2) are cultured at 20-32℃ for 4-40 hours. After the culture is completed, they are washed with tap water and drained. The drained soybeans are then called dry soybeans.

[0010] (4) After mixing the drained soybeans from step (3) with water, grind them into a paste, separate the paste from the residue, and collect the soybean paste liquid;

[0011] (5) Steam the soybean milk at 100-135℃ for 10s-60min, and after steaming, cool it to room temperature to obtain the steamed soybean milk.

[0012] (6) The cooked soybean slurry obtained in step (5) is hydrolyzed with cellulase for 30-60 min under natural pH and temperature conditions of 30-60℃, wherein the amount of cellulase added is 10-40 U / g of dry soybeans; then the pH of the soybean slurry is adjusted to 8.0-10.0 with NaOH solution; after adjusting the temperature to 35-60℃, alkaline protease is added to continue the enzymatic hydrolysis for 1-6 h, wherein the amount of alkaline protease added is 10-40 U / g of dry soybeans. After the enzymatic hydrolysis is completed, the enzyme is inactivated by heating, the hydrolysate is collected and centrifuged, and the supernatant is collected after centrifugation.

[0013] (7) The supernatant obtained in step (6) is concentrated and dried to obtain selenium-enriched soybean enzymatic hydrolysate; the total selenium content of the selenium-enriched soybean enzymatic hydrolysate is 5.98-30.13 mg / kg, and the organic selenium content accounts for 95.12%-99.63%.

[0014] Preferably, in step (2), the selenium content in the selenium-enriched nutrient solution is 20-30 mg / L, and the selenium is derived from sodium selenite or nano-selenium; the contents of vitamin C, vitamin B1, and vitamin B7 are 1500-2500 mg / L, 10-15 mg / L, and 50-70 μg / L, respectively; the ultrasonic treatment during soaking is performed during 0-8 hours of soaking; the conditions for ultrasonic treatment are: ultrasonic frequency of 40-60 kHz and ultrasonic intensity density of 0.3-0.8 W / cm³. 2 The ultrasound time is 15-25 minutes.

[0015] Preferably, the temperature for soybean cultivation in step (3) is 25-30℃ and the time is 20-30h.

[0016] Preferably, in step (4), the drained soybeans are mixed with water, and the weight ratio of water to soybeans is 2-6:1.

[0017] Preferably, the temperature for steaming the soybean slurry in step (5) is 120-130℃ and the time is 3-30 minutes.

[0018] Preferably, in step (6), the temperature for adding cellulase for enzymatic hydrolysis is 45-55℃, the time is 40-50 min, and the amount of cellulase added is 20-30 U / g of dried soybeans based on the weight of dried soybeans; the pH value of the soybean slurry is 8.5-9.5; the temperature for adding alkaline protease for continued enzymatic hydrolysis is 45-55℃, the hydrolysis time is 3-4 h, and the amount of alkaline protease added is 25-35 U / g of dried soybeans based on the weight of dried soybeans; the temperature for heating enzymatic hydrolysis is 100℃, the heating time is 5-10 min; and the centrifugation conditions are centrifugation at 5000g centrifugal force for 10 min.

[0019] Furthermore, the total selenium content of the selenium-enriched soybean enzymatic hydrolysate is 10.10-15.26 mg / kg, and the proportion of organic selenium is 97.86%-98.12%.

[0020] The selenium-enriched soybean enzymatic hydrolysate prepared in this invention has applications in the preparation of products that lower triglyceride, cholesterol, low-density lipoprotein and blood glucose levels, and increase high-density lipoprotein.

[0021] A product characterized in that it comprises the selenium-enriched soybean enzymatic hydrolysate, the product comprising powder, granule, pill, capsule, oral liquid or tablet.

[0022] The product is used in the preparation of pharmaceuticals that lower triglyceride, cholesterol, low-density lipoprotein and blood glucose levels and increase high-density lipoprotein.

[0023] For adults, daily intake of 0.085-2.0g / kg body weight of selenium-enriched soybean hydrolysate can effectively reduce triglyceride, cholesterol, low-density lipoprotein and blood sugar levels in patients with hyperlipidemia and hyperglycemia, and increase high-density lipoprotein levels in patients with hyperlipidemia.

[0024] The beneficial effects of this invention are:

[0025] (1) The selenium-enriched soybean enzymatic hydrolysate produced by this invention has a high organic selenium content and has a significant ability to lower blood lipids and blood sugar. Moreover, the selenium-enriched soybean enzymatic hydrolysate produced is safe and has a short production cycle.

[0026] (2) Based on extensive experiments, this invention rationally designs the components and proportions of selenium-enriched nutrient solution, cleverly utilizes ultrasonic technology, and employs ultrasonic treatment in conjunction with vitamins to promote the absorption and conversion of inorganic selenium into organic selenium in soybeans. Simultaneously, by combining specific ultrasonic and enzymatic hydrolysis conditions, it significantly improves the selenium-enriching capacity of soybeans and shortens the production time of selenium-enriched soybeans, thus developing a selenium-enriched soybean enzymatic hydrolysate with lipid-lowering and blood sugar-lowering functions. Compared with the production of selenium-enriched soybean enzymatic hydrolysates from selenium-enriched soybeans, this invention has advantages such as a shorter production cycle, no seasonal influence, high selenium content in the hydrolysate, and low cost. Furthermore, the product exhibits significant lipid-lowering and blood sugar-lowering functions, achieving unexpectedly significant results. Therefore, this invention has important economic and social value. Attached Figure Description

[0027] Figure 1 The effect of soaking time on the total selenium, organic selenium content and organic selenium conversion rate in soybean enzymatic hydrolysate.

[0028] Figure 2 The effect of selenium content in selenium-enriched nutrient solution on total selenium, organic selenium content and organic selenium conversion rate in soybean enzymatic hydrolysate.

[0029] Figure 3 The effect of vitamin C content in selenium-enriched nutrient solution on total selenium, organic selenium content and organic selenium conversion rate in soybean enzymatic hydrolysate.

[0030] Figure 4 The effect of VB1 in selenium-enriched nutrient solution on the total selenium, organic selenium content and organic selenium conversion rate of soybean enzymatic hydrolysate.

[0031] Figure 5 The effect of VB7 in selenium-enriched nutrition on the total selenium, organic selenium content and organic selenium conversion rate in soybean enzymatic hydrolysate.

[0032] Figure 6 The effect of ultrasonic frequency on the total selenium, organic selenium content and organic selenium conversion rate in soybean enzymatic hydrolysate.

[0033] Figure 7The effect of ultrasonic intensity density on the total selenium, organic selenium content and organic selenium conversion rate of soybean enzymatic hydrolysate.

[0034] Figure 8 The effect of ultrasound time on the total selenium, organic selenium content and organic selenium conversion rate of soybean enzymatic hydrolysate.

[0035] Figure 9 To investigate the effects of cultivation temperature on the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate.

[0036] Figure 10 To investigate the effects of cultivation time on the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate.

[0037] Figure 11 The effect of the water-to-soybean weight ratio on the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate was investigated.

[0038] Figure 12 The effect of cooking time of soybean slurry on the total selenium, organic selenium content and organic selenium conversion rate in soybean enzymatic hydrolysate.

[0039] Figure 13 The effect of cooking temperature of soybean slurry on the total selenium, organic selenium content and organic selenium conversion rate in soybean enzymatic hydrolysate.

[0040] Figure 14 The effect of cellulase hydrolysis temperature on the total selenium, organic selenium content and organic selenium conversion rate of soybean hydrolysate.

[0041] Figure 15 The effect of cellulase hydrolysis time on the total selenium, organic selenium content and organic selenium conversion rate of soybean hydrolysate.

[0042] Figure 16 The effect of cellulase addition on total selenium, organic selenium content and organic selenium conversion rate in soybean enzymatic hydrolysate.

[0043] Figure 17 The effect of pH on the total selenium, organic selenium content and organic selenium conversion rate of soybean hydrolysate by alkaline protease hydrolysis.

[0044] Figure 18 The effect of alkaline protease hydrolysis temperature on the total selenium, organic selenium content, and organic selenium conversion rate of soybean hydrolysate was investigated.

[0045] Figure 19 The effect of alkaline protease hydrolysis time on the total selenium, organic selenium content and organic selenium conversion rate of soybean hydrolysate.

[0046] Figure 20 The effect of alkaline protease addition on total selenium, organic selenium content and organic selenium conversion rate of soybean hydrolysate. Detailed Implementation

[0047] The specific embodiments of the present invention will be further described below with reference to examples, but the implementation and protection scope of the present invention are not limited thereto.

[0048] The methods for determining the total selenium, organic selenium, and inorganic selenium content involved in this invention refer to GB 5009.93-2017 and the method of Jing Dawei et al. (Selenium enrichment, fruit quality and yield of winter jujubeas affected by addition of sodium selenite[J].Scientia Horticulturae,2017,225,1-5). The organic selenium content is the total selenium content minus the inorganic selenium content.

[0049] Note: The amount of enzyme added in this invention is based on the mass of soybeans (denoted as dry soybeans) after the culture is completed and the water has been drained.

[0050] I. An Investigation into Soybean Soaking Time

[0051] Wash the soybeans with clean water; then soak them in a selenium-enriched nutrient solution. The selenium content, vitamin C, vitamin B1, and vitamin B7 in the selenium-enriched nutrient solution are 30 mg / L, 2000 mg / L, 12.5 mg / L, and 60 μg / L, respectively. After soaking the soybeans for 8 hours, perform ultrasonic treatment under the following conditions: ultrasonic frequency 45 kHz, ultrasonic intensity density 0.6 W / cm³. 2 Soybeans were ultrasonically soaked for 20 minutes and then cultured at 27°C for 25 hours. After culture, they were washed with tap water and drained. The drained soybeans were then labeled as dry soybeans. The drained soybeans were mixed with three times their weight of water and ground using a grinder. The resulting soybean slurry was heated at 125°C for 15 minutes. The soybean slurry was then hydrolyzed with cellulase at its natural pH and 50°C for 45 minutes. The amount of cellulase added was 25 U / g of dry soybeans. Then, the pH of the soybean slurry was adjusted to 9.0 and the temperature to 50°C with NaOH solution. Alkaline protease was added and hydrolysis continued for 3.5 hours. The amount of alkaline protease added was 30 U / g of dry soybeans. After hydrolysis, the enzyme was inactivated by heating at 100°C for 5 minutes. The hydrolysate was centrifuged at 5000g for 10 minutes. The supernatant was collected, and the precipitate was discarded. The supernatant was concentrated and dried to obtain selenium-enriched soybean hydrolysate.

[0052] Figure 1 The effect of soaking time on the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate; by Figure 1It was found that as the soaking time of soybeans increased, the total selenium, organic selenium content, and organic selenium conversion rate in the soybean enzymatic hydrolysate gradually increased (reaching the highest at 25 hours). At 8 hours of soaking, the total selenium and organic selenium content in the soybean enzymatic hydrolysate were significantly higher than in the initial stage. This is related to the swelling of soybeans after water absorption, the increase in cell wall pores, and the need for a large amount of nutrients for soybean germination. However, excessively long soaking times also lead to a decrease in the total selenium, organic selenium content, and organic selenium conversion rate in the soybean enzymatic hydrolysate. In particular, when soybeans were soaked for more than 30 hours, the total selenium, organic selenium content, and organic selenium conversion rate all decreased significantly. This is related to the accumulation of excessive selenium in the soybeans, which inhibits normal metabolic activity, preventing the soybeans from absorbing selenium from the solution or converting inorganic selenium into organic selenium.

[0053] II. Investigation of Selenium Content in Selenium-Enriched Nutrient Solution

[0054] Soybeans were rinsed with clean water; after rinsing, they were soaked in a selenium-enriched nutrient solution; the selenium content was varied by controlling the content of vitamin C, vitamin B1, and vitamin B7 in the selenium-enriched nutrient solution to 2000 mg / L, 12.5 mg / L, and 60 μg / L, respectively; the soaking time in the selenium-enriched nutrient solution was 25 h; and the soybeans were sonicated after 8 h of soaking under the following conditions: ultrasonic frequency 45 kHz and ultrasonic intensity density 0.6 W / cm³. 2 The process involved: 1. Ultrasonic treatment for 20 minutes; 2. Soybeans were cultured at 27℃ for 25 hours, then washed with tap water and drained; the drained soybeans were then labeled as dried soybeans. The drained soybeans were mixed with three times their weight of water and ground using a grinder. The soybean slurry was heated at 125℃ for 15 minutes. The soybean slurry was then enzymatically hydrolyzed for 45 minutes at its natural pH and 50℃ with the addition of cellulase (25 U / g dried soybeans). The pH of the soybean slurry was then adjusted to 9.0 and the temperature to 50℃ using NaOH solution. Alkaline protease was added and hydrolysis continued for 3.5 hours (30 U / g dried soybeans). After hydrolysis, the enzyme was inactivated by heating at 100℃ for 5 minutes. The hydrolysate was centrifuged at 5000g for 10 minutes, and the supernatant was collected, discarding the precipitate. The supernatant was then concentrated and dried to obtain selenium-enriched soybean hydrolysate.

[0055] Figure 2 The effect of selenium content in selenium-enriched nutrient solution on total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate; by Figure 2It can be seen that as the selenium content of the selenium-enriched nutrient solution increases, the total selenium and organic selenium content in the soybean enzymatic hydrolysate gradually increase (reaching its highest level at 60 mg / L). However, excessively high selenium content in the selenium-enriched nutrient solution (>60 mg / L) also leads to a decrease in the total selenium and organic selenium content in the soybean enzymatic hydrolysate. Notably, as the selenium content of the selenium-enriched nutrient solution increases, the conversion rate of organic selenium in the soybean enzymatic hydrolysate continuously decreases. These phenomena indicate that soybeans can effectively absorb inorganic selenium and convert it into organic selenium in a low-selenium environment. However, excessively high selenium content in the environment can disrupt the normal metabolic activities of soybeans, preventing them from continuing to absorb selenium from the solution or converting inorganic selenium into organic selenium.

[0056] III. Investigation into the Vitamin C Content in Selenium-Enriched Nutrient Solution

[0057] Wash soybeans with clean water; then soak them in a selenium-enriched nutrient solution; control the selenium, vitamin B1, and vitamin B7 content in the selenium-enriched nutrient solution to 30 mg / L, 12.5 mg / L, and 60 μg / L respectively, and vary the vitamin C content; soak the soybeans in the selenium-enriched nutrient solution for 25 hours, and then sonicate them after 8 hours of soaking under the following conditions: ultrasonic frequency 45 kHz and ultrasonic intensity density 0.6 W / cm³. 2 The process involved: 1. Ultrasonic treatment for 20 minutes; 2. Soybeans were cultured at 27℃ for 25 hours, then washed with tap water and drained. The drained soybeans were then labeled as dried soybeans. The drained soybeans were mixed with three times their weight of water and ground using a grinder. The soybean slurry was heated at 125℃ for 15 minutes. The soybean slurry was then enzymatically hydrolyzed for 45 minutes at its natural pH and 50℃ with the addition of cellulase (25 U / g dried soybeans). The pH of the soybean slurry was then adjusted to 9.0 and the temperature to 50℃ using NaOH solution. Alkaline protease was added and hydrolysis continued for 3.5 hours (30 U / g dried soybeans). After hydrolysis, the enzyme was inactivated by heating at 100℃ for 5 minutes. The hydrolysate was centrifuged at 5000g for 10 minutes, and the supernatant was collected, discarding the precipitate. The supernatant was then concentrated and dried to obtain selenium-enriched soybean hydrolysate.

[0058] Figure 3 The effect of vitamin C content in selenium-enriched nutrient solution on the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate; by Figure 3It was found that with the increase of vitamin C added to the selenium-enriched nutrient solution, the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate gradually increased (reaching the highest at 2000 mg / L). In particular, when the vitamin C addition increased from 0 to 500 mg / L, the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate significantly increased, which is related to vitamin C's ability to resist stress (induced by high concentrations of selenium). However, excessive vitamin C addition also led to a decrease in the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate. When the vitamin C addition exceeded 5000 mg / L, the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate all decreased significantly. This is related to the accumulation of excessive vitamin C in soybeans, which disrupted the normal metabolic activities of soybeans, preventing them from absorbing selenium from the solution and from converting inorganic selenium into organic selenium.

[0059] IV. Investigation of VB1 content in selenium-enriched nutrient solution

[0060] Wash soybeans with clean water; then soak them in a selenium-enriched nutrient solution; control the selenium, vitamin C, and vitamin B7 content in the selenium-enriched nutrient solution to 30 mg / L, 2000 mg / L, and 60 μg / L respectively, and vary the vitamin B1 content; soak the soybeans in the selenium-enriched nutrient solution for 25 hours, and then sonicate them after 8 hours of soaking under the following conditions: ultrasonic frequency 45 kHz and ultrasonic intensity density 0.6 W / cm³. 2 The process involved: 1. Ultrasonic treatment for 20 minutes; 2. Soybeans were cultured at 27℃ for 25 hours, then washed with tap water and drained. The drained soybeans were then labeled as dried soybeans. The drained soybeans were mixed with three times their weight of water and ground using a grinder. The soybean slurry was heated at 125℃ for 15 minutes. The soybean slurry was then enzymatically hydrolyzed for 45 minutes at its natural pH and 50℃ with the addition of cellulase (25 U / g dried soybeans). The pH of the soybean slurry was then adjusted to 9.0 and the temperature to 50℃ using NaOH solution. Alkaline protease was added and hydrolysis continued for 3.5 hours (30 U / g dried soybeans). After hydrolysis, the enzyme was inactivated by heating at 100℃ for 5 minutes. The hydrolysate was centrifuged at 5000g for 10 minutes, and the supernatant was collected, discarding the precipitate. The supernatant was then concentrated and dried to obtain selenium-enriched soybean hydrolysate.

[0061] Figure 4 The effect of vitamin B1 in selenium-enriched nutrient solution on the total selenium, organic selenium content, and organic selenium conversion rate of soybean enzymatic hydrolysate; Figure 4It is known that with the increase of VB1 addition in selenium-enriched nutrient solution, the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate gradually increased. In particular, when the VB1 addition reached 12.5 mg / L, the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate significantly increased, which is related to the anti-stress (induced by high concentration of selenium) ability of VB1. However, excessive VB1 addition can also lead to a decrease in the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate; more is not necessarily better. Especially when the VB1 addition is greater than 20 mg / L, the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate all significantly decreased. This is related to the accumulation of excessive VB1 in soybeans, which disrupts the normal metabolic activities of soybeans, preventing them from absorbing selenium from the solution or converting inorganic selenium into organic selenium.

[0062] V. Investigation into the VB7 content in selenium-enriched nutrient solution

[0063] Wash soybeans with clean water; then soak them in selenium-enriched nutrient solution; control the selenium, vitamin C, and vitamin B1 content in the selenium-enriched nutrient solution to 30 mg / L, 2000 mg / L, and 12.5 mg / L respectively, and adjust the VB7 content; soak soybeans in the selenium-enriched nutrient solution for 25 hours, and sonicate them after 8 hours of soaking under the following conditions: ultrasonic frequency 45 kHz, ultrasonic intensity density 0.6 W / cm2, and ultrasonic time 20 min; incubate soybeans at 27℃ for 25 hours, then wash them with tap water and drain them; the drained soybeans are then labeled as dried soybeans; mix the drained soybeans with 3 times their weight of water and grind them... Soybean pulp was ground using a pulping machine; the soybean pulp was heated at 125℃ for 15 minutes; the soybean pulp was then enzymatically hydrolyzed for 45 minutes at its natural pH and 50℃ with the addition of cellulase at a dosage of 25 U / g dry soybeans. The pH of the soybean pulp was then adjusted to 9.0 and the temperature to 50℃ using NaOH solution, and alkaline protease was added to continue the enzymatic hydrolysis for 3.5 hours at a dosage of 30 U / g dry soybeans. After hydrolysis, the enzyme was inactivated by heating at 100℃ for 5 minutes. The hydrolysate was then centrifuged at 5000g for 10 minutes, and the supernatant was collected, discarding the precipitate. The supernatant was concentrated and dried to obtain selenium-enriched soybean hydrolysate.

[0064] Figure 5 The effect of VB7 in selenium-enriched nutrition on the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate; by Figure 5It was found that with the increase of VB7 addition in the selenium-enriched nutrient solution, the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate gradually increased. In particular, when the VB7 addition reached 60 μg / L, the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate significantly increased, which is related to the ability of VB7 to promote protein metabolism. However, excessive VB7 addition also led to a decrease in the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate. In particular, when the VB7 addition was greater than 100 μg / L, the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate all decreased significantly. This is related to the accumulation of excessive VB7 in soybeans, which disrupted the normal metabolic activities of soybeans, making it impossible for soybeans to continue absorbing selenium from the solution or converting inorganic selenium into organic selenium.

[0065] VI. Investigation of Ultrasonic Frequency

[0066] Wash the soybeans with clean water; then soak them in a selenium-enriched nutrient solution. The selenium, vitamin C, vitamin B1, and vitamin B7 content in the selenium-enriched nutrient solution were 30 mg / L, 2000 mg / L, 12.5 mg / L, and 60 μg / L, respectively. The soybeans were soaked in the selenium-enriched nutrient solution for 25 hours. Ultrasonication was started 8 hours after soaking, with the ultrasonic intensity density at 0.6 W / cm³. 2 The process involved sonication for 20 minutes, with variations in the sonic frequency. Soybeans were cultured at 27℃ for 25 hours, then washed with tap water and drained. These drained soybeans were then labeled as dried soybeans. The drained soybeans were mixed with three times their weight of water and ground using a grinder. The soybean slurry was heated at 125℃ for 15 minutes. The slurry was then enzymatically hydrolyzed for 45 minutes at its natural pH and 50℃ with the addition of cellulase (25 U / g dried soybeans). The pH was then adjusted to 9.0 and the temperature to 50℃ using NaOH solution, followed by the addition of alkaline protease (30 U / g dried soybeans) for another 3.5 hours. After hydrolysis, the enzyme was inactivated by heating at 100℃ for 5 minutes. The hydrolysate was then centrifuged at 5000g for 10 minutes, and the supernatant was collected, discarding the precipitate. The supernatant was concentrated and dried to obtain the selenium-enriched soybean hydrolysate.

[0067] Figure 6 The effects of ultrasonic frequency on the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate; by Figure 6It can be seen that with the increase of ultrasonic frequency, the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate gradually increase. In particular, when the ultrasonic frequency increases from 18 kHz to 45 kHz, the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate significantly increase, which is related to the ability of appropriate ultrasound to promote plant seed germination. However, excessively high ultrasonic frequency can also lead to a decrease in the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate. When the ultrasonic frequency is greater than 65 kHz, the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate all decrease significantly. Soybeans can neither continue to absorb selenium from the solution nor continue to convert inorganic selenium into organic selenium.

[0068] VII. Investigation of Ultrasonic Intensity and Density

[0069] Wash soybeans with clean water; then soak them in a selenium-enriched nutrient solution. The selenium, vitamin C, vitamin B1, and vitamin B7 content in the solution are 30 mg / L, 2000 mg / L, 12.5 mg / L, and 60 μg / L, respectively. Soak the soybeans in the solution for 25 hours. After 8 hours of soaking, begin sonication at a frequency of 45 kHz for 20 minutes, varying the intensity and density. Incubate the soybeans at 27℃ for 25 hours. After incubation, wash them with tap water and drain. The drained soybeans are then labeled as dried soybeans. Mix the drained soybeans with three times their weight of water and grind them. Soybean pulp was ground using a pulping machine; the soybean pulp was heated at 125℃ for 15 minutes; the soybean pulp was then enzymatically hydrolyzed for 45 minutes at its natural pH and 50℃ with the addition of cellulase at a dosage of 25 U / g dry soybeans. The pH of the soybean pulp was then adjusted to 9.0 and the temperature to 50℃ using NaOH solution, and alkaline protease was added to continue the enzymatic hydrolysis for 3.5 hours at a dosage of 30 U / g dry soybeans. After hydrolysis, the enzyme was inactivated by heating at 100℃ for 5 minutes. The hydrolysate was then centrifuged at 5000g for 10 minutes, and the supernatant was collected, discarding the precipitate. The supernatant was concentrated and dried to obtain selenium-enriched soybean hydrolysate.

[0070] Figure 7 The effect of ultrasonic intensity density on the total selenium, organic selenium content, and organic selenium conversion rate of soybean enzymatic hydrolysate; by Figure 7 It can be seen that with the increase of ultrasonic intensity and density, the total selenium, organic selenium content and organic selenium conversion rate in soybean enzymatic hydrolysate gradually increase, especially when the ultrasonic intensity reaches 0.6 W / cm². 2 The total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate were significantly increased, which is related to the ability of appropriate ultrasound to promote plant seed germination. However, excessively high ultrasound intensity can also lead to a decrease in the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate, especially when the ultrasound frequency is greater than 1.5 W / cm. 2At that time, the total selenium, organic selenium content and organic selenium conversion rate in soybean enzymatic hydrolysate all decreased significantly. Soybeans could neither continue to absorb selenium from the solution nor continue to convert inorganic selenium into organic selenium.

[0071] VIII. Investigation of Ultrasonic Time

[0072] Wash soybeans with clean water; then soak them in a selenium-enriched nutrient solution. The selenium, vitamin C, vitamin B1, and vitamin B7 content in the selenium-enriched nutrient solution are 30 mg / L, 2000 mg / L, 12.5 mg / L, and 60 μg / L, respectively. The soybeans are soaked in the selenium-enriched nutrient solution for 25 hours. After 8 hours of soaking, the soybeans are sonicated under the following conditions: ultrasonic frequency of 45 kHz and ultrasonic intensity density of 0.6 W / cm³. 2 The ultrasonic time was changed; soybeans were cultured at 27℃ for 25 hours, and after the culture was completed, they were washed with tap water and drained; the drained soybeans were then recorded as dry soybeans; the drained soybeans were mixed with 3 times their weight of water and ground into a paste using a grinder; the soybean paste was heated at 125℃ for 15 minutes; the soybean paste was then enzymatically hydrolyzed for 45 minutes at its natural pH and 50℃ with the addition of cellulase at a dosage of 25 U / g dry soybeans, and then the pH of the soybean paste was adjusted to 9.0 and the temperature to 50℃ with NaOH solution, and alkaline protease was added to continue the enzymatic hydrolysis for 3.5 hours at a dosage of 30 U / g dry soybeans; after the enzymatic hydrolysis was completed, the enzyme was inactivated by heating at 100℃ for 5 minutes, and the hydrolysate was centrifuged at 5000g for 10 minutes, the supernatant was collected, and the precipitate was discarded; the supernatant was concentrated and dried to obtain selenium-enriched soybean hydrolysate.

[0073] Figure 8 The effect of ultrasound time on the total selenium, organic selenium content, and organic selenium conversion rate of soybean enzymatic hydrolysate; by Figure 8 It was found that with the extension of ultrasonic time, the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate gradually increased. In particular, when the ultrasonic time increased from 0 to 20 minutes, the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate significantly increased, which is related to the ability of appropriate ultrasound to promote plant seed germination. However, excessively long ultrasonic time also led to a decrease in the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate. Especially when the ultrasonic frequency exceeded 30 minutes, the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate all decreased significantly, indicating that soybeans could neither continue to absorb selenium from the solution nor continue to convert inorganic selenium into organic selenium.

[0074] As shown in Table 1, Experiments 1 to 8, vitamins Vc, VB1, and VB7 not only promote the absorption of selenium in soybeans and its conversion into organic selenium, but also, through synergistic effects, further promote the absorption of selenium and its conversion into organic selenium in soybeans. As shown in Table 1, Experiments 9 to 16, ultrasound, as well as single and multiple vitamins, can all, through synergistic effects, further promote the absorption of selenium in soybeans and its conversion into organic selenium.

[0075] Table 1. Effects of synergistic effect on total selenium, organic selenium content and organic selenium conversion rate of selenium-enriched soybean enzymatic hydrolysate.

[0076]

[0077]

[0078] Note: 1. Data in the same column without identical letters indicates a significant difference between the numbers (p<0.05); 2. For experiments 1-16, the types of added nutrients and ultrasonic conditions gradually increased with the serial number, while other experimental conditions remained the same, as follows: Soybeans were soaked for 25 hours, cultured at 27℃ for 25 hours, washed with tap water after culture, and drained; the drained soybeans were then recorded as dry soybeans; the drained soybeans were mixed with 3 times their weight of water and ground into a paste using a grinder; the soybean paste was heated at 125℃ for 15 minutes; the soybean paste was enzymatically hydrolyzed with cellulase at its natural pH and 50℃ for 45 minutes, with a cellulase addition of 25 U / g dry soybeans; then the pH of the soybean paste was adjusted to 9.0 and the temperature to 50℃ using NaOH solution, and alkaline protease was added to continue enzymatic hydrolysis for 3.5 hours, with an alkaline protease addition of 30 U / g dry soybeans. After enzymatic hydrolysis, the enzyme was inactivated by heating at 100℃ for 5 minutes. The hydrolysate was centrifuged at 5000g for 10 minutes. The supernatant was collected and the precipitate was discarded. The supernatant was concentrated and dried to obtain selenium-enriched soybean hydrolysate.

[0079] IX. An Investigation into the Cultivation Temperature of Soybeans

[0080] Wash soybeans with clean water; then soak them in a selenium-enriched nutrient solution. The selenium, vitamin C, vitamin B1, and vitamin B7 content in the selenium-enriched nutrient solution are 30 mg / L, 2000 mg / L, 12.5 mg / L, and 60 μg / L, respectively. The soybeans are soaked in the selenium-enriched nutrient solution for 25 hours. After 8 hours of soaking, the soybeans are sonicated under the following conditions: ultrasonic frequency of 45 kHz and ultrasonic intensity density of 0.6 W / cm³. 2The ultrasonic treatment time was 20 minutes; the soybean cultivation time was controlled at 25 hours, with varying cultivation temperature. After cultivation, the soybeans were washed with tap water and drained. The drained soybeans were then labeled as dry soybeans. The drained soybeans were mixed with three times their weight of water and ground using a grinder. The soybean slurry was heated at 125°C for 15 minutes. The soybean slurry was then hydrolyzed for 45 minutes with cellulase at its natural pH and 50°C, with a cellulase addition of 25 U / g dry soybeans. The pH of the soybean slurry was then adjusted to 9.0 and the temperature to 50°C using NaOH solution. Alkaline protease was added and hydrolysis continued for 3.5 hours, with an alkaline protease addition of 30 U / g dry soybeans. After hydrolysis, the enzyme was inactivated by heating at 100°C for 5 minutes. The hydrolysate was centrifuged at 5000g for 10 minutes, and the supernatant was collected, discarding the precipitate. The supernatant was concentrated and dried to obtain selenium-enriched soybean hydrolysate.

[0081] Figure 9 To investigate the effects of different culture temperatures on the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysates; Figure 9 It can be seen that as the soybean cultivation temperature increases, the total selenium, organic selenium content, and organic selenium conversion rate in the soybean enzymatic hydrolysate gradually increase (reaching their highest at 27℃). Particularly when the cultivation temperature increases from 17℃ to 20℃, the total selenium, organic selenium content, and organic selenium conversion rate in the soybean enzymatic hydrolysate significantly increase, which is related to the optimal growth temperature for soybean germination. However, excessively high cultivation temperatures also cause the total selenium, organic selenium content, and organic selenium conversion rate in the soybean enzymatic hydrolysate to cease increasing. When the cultivation temperature exceeds 32℃, the total selenium, organic selenium content, and organic selenium conversion rate in the soybean enzymatic hydrolysate rapidly begin to decline, indicating that the soybean can no longer absorb selenium from the solution, nor can it continue to convert inorganic selenium into organic selenium.

[0082] 10. An Investigation into the Cultivation Time of Soybeans

[0083] Wash soybeans with clean water; then soak them in a selenium-enriched nutrient solution. The selenium, vitamin C, vitamin B1, and vitamin B7 content in the selenium-enriched nutrient solution were 30 mg / L, 2000 mg / L, 12.5 mg / L, and 60 μg / L, respectively. The soybeans were soaked in the selenium-enriched nutrient solution for 25 hours. After 8 hours of soaking, the soybeans were sonicated under the following conditions: ultrasonic frequency of 45 kHz and ultrasonic intensity density of 0.6 W / cm³. 2The ultrasonic treatment time was 20 minutes; the soybean cultivation temperature was controlled at 27℃, and the cultivation time was varied. After cultivation, the soybeans were washed with tap water and drained. The drained soybeans were then labeled as dry soybeans. The drained soybeans were mixed with 3 times their weight of water and ground using a grinder. The soybean slurry was heated at 125℃ for 15 minutes. The soybean slurry was then enzymatically hydrolyzed for 45 minutes at its natural pH and 50℃ with the addition of cellulase at a dosage of 25 U / g of dry soybeans. The pH of the soybean slurry was then adjusted to 9.0 and the temperature to 50℃ with NaOH solution. Alkaline protease was added and the enzymatic hydrolysis continued for 3.5 hours at a dosage of 30 U / g of dry soybeans. After the enzymatic hydrolysis was completed, the enzyme was inactivated by heating at 100℃ for 5 minutes. The hydrolysate was centrifuged at 5000g for 10 minutes, and the supernatant was collected and the precipitate was discarded. The supernatant was concentrated and dried to obtain selenium-enriched soybean hydrolysate.

[0084] Figure 10 To investigate the effects of cultivation time on the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysates; Figure 10 It can be seen that as the soybean cultivation time increases, the total selenium, organic selenium content, and organic selenium conversion rate in the soybean enzymatic hydrolysate gradually increase. When the cultivation time increases from 2 hours to 25 hours, the total selenium, organic selenium content, and organic selenium conversion rate in the soybean enzymatic hydrolysate significantly increase, which is related to the continuous improvement of the absorption, metabolism, and conversion capacity of various nutrients during soybean germination. However, excessively long cultivation time will also cause the total selenium, organic selenium content, and organic selenium conversion rate in the soybean enzymatic hydrolysate to no longer increase. In particular, when the cultivation time exceeds 40 hours, the total selenium, organic selenium content, and organic selenium conversion rate in the soybean enzymatic hydrolysate begin to decrease significantly, and the soybean can neither continue to absorb selenium from the solution nor continue to convert inorganic selenium into organic selenium.

[0085] XI. Investigation into the weight ratio of water to soybeans

[0086] Wash soybeans with clean water; then soak them in a selenium-enriched nutrient solution. The selenium, vitamin C, vitamin B1, and vitamin B7 content in the selenium-enriched nutrient solution are 30 mg / L, 2000 mg / L, 12.5 mg / L, and 60 μg / L, respectively. The soybeans are soaked in the selenium-enriched nutrient solution for 25 hours. After 8 hours of soaking, the soybeans are sonicated under the following conditions: ultrasonic frequency of 45 kHz and ultrasonic intensity density of 0.6 W / cm³. 2The ultrasonic treatment time was 20 min; the soybean cultivation temperature was 27℃ and the cultivation time was 25 h; after cultivation, the soybeans were washed with tap water and drained; the drained soybeans were then labeled as dry soybeans; the drained soybeans were mixed with water and ground into a slurry, and the resulting soybean slurry was heated at 125℃ for 15 min; the soybean slurry was then hydrolyzed with cellulase at its natural pH and 50℃ for 45 min, with a cellulase addition of 25 U / g dry soybeans; then the pH of the soybean slurry was adjusted to 9.0 and the temperature to 50℃ with NaOH solution, and alkaline protease was added to continue the hydrolysis for 3.5 h, with an alkaline protease addition of 30 U / g dry soybeans; after the hydrolysis was completed, the enzyme was inactivated by heating at 100℃ for 5 min, and the hydrolysate was centrifuged at 5000g for 10 min, the supernatant was collected, and the precipitate was discarded; the supernatant was concentrated and dried to obtain selenium-enriched soybean hydrolysate.

[0087] Figure 11 The effect of the water-to-soybean weight ratio on the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate; by Figure 11 It can be seen that as the weight ratio of water to soybeans increases, the total selenium, organic selenium content, and organic selenium conversion rate in the soybean enzymatic hydrolysate gradually increase. In particular, when the weight ratio of water to soybeans is 3, the total selenium, organic selenium content, and organic selenium conversion rate in the soybean enzymatic hydrolysate increase significantly. This is related to the decrease in solid content during the subsequent enzymatic hydrolysis process, which is conducive to solid dissolution and enzymatic hydrolysis. However, when the weight ratio of water to soybeans is greater than 3, the total selenium, organic selenium content, and organic selenium conversion rate in the soybean enzymatic hydrolysate no longer increase, and it also increases the subsequent drying cost. Therefore, a weight ratio of water to soybeans of 3 is optimal.

[0088] XII. An Investigation into the Heating Time of Soybean Milk

[0089] Wash soybeans with clean water; then soak them in a selenium-enriched nutrient solution. The selenium, vitamin C, vitamin B1, and vitamin B7 content in the selenium-enriched nutrient solution are 30 mg / L, 2000 mg / L, 12.5 mg / L, and 60 μg / L, respectively. The soybeans are soaked in the selenium-enriched nutrient solution for 25 hours. After 8 hours of soaking, the soybeans are sonicated under the following conditions: ultrasonic frequency of 45 kHz and ultrasonic intensity density of 0.6 W / cm³. 2The ultrasonic treatment time was 20 minutes; the soybean culture temperature was 27℃ and the culture time was 25 hours. After the culture was completed, the soybeans were washed with tap water and drained. The drained soybeans were then labeled as dry soybeans. The drained soybeans were mixed with 3 times their weight of water and ground into a paste using a grinder. The soybean paste was heated and cooked at 125℃. After cooking, the paste was cooled to room temperature. Cellulase was added to the soybean paste at its natural pH and 50℃ for 45 minutes for enzymatic hydrolysis. The amount of cellulase added was 25 U / g of dry soybeans. Then, the pH of the soybean paste was adjusted to 9.0 and the temperature was adjusted to 50℃ with NaOH solution. Alkaline protease was added and enzymatic hydrolysis continued for 3.5 hours. The amount of alkaline protease added was 30 U / g of dry soybeans. After the enzymatic hydrolysis was completed, the paste was heated at 100℃ for 5 minutes to inactivate the enzyme. The hydrolysate was centrifuged at 5000g for 10 minutes. The supernatant was collected and the precipitate was discarded. The supernatant was concentrated and dried to obtain selenium-enriched soybean hydrolysate.

[0090] Figure 12 The effect of cooking time of soybean slurry on the total selenium, organic selenium content and organic selenium conversion rate in soybean enzymatic hydrolysate; by Figure 12 It can be seen that as the heating time of soybean slurry increases, the total selenium, organic selenium content, and organic selenium conversion rate in the soybean enzymatic hydrolysate gradually increase. In particular, when the heating time reaches 15 minutes, the total selenium, organic selenium content, and organic selenium conversion rate in the soybean enzymatic hydrolysate increase significantly. This is related to the fact that heating causes moderate denaturation of soybeans in the slurry, which is beneficial for subsequent enzymatic hydrolysis and dissolution. However, after the heating time of the slurry exceeds 15 minutes, the total selenium, organic selenium content, and organic selenium conversion rate in the soybean enzymatic hydrolysate no longer increase and even decrease slightly. This is related to the fact that excessive heating causes excessive denaturation and precipitation of proteins (the main carriers of organic selenium) in the slurry.

[0091] XIII. An Investigation into the Heating Temperature of Soybean Slurry

[0092] Wash soybeans with clean water; then soak them in a selenium-enriched nutrient solution. The selenium, vitamin C, vitamin B1, and vitamin B7 content in the selenium-enriched nutrient solution are 30 mg / L, 2000 mg / L, 12.5 mg / L, and 60 μg / L, respectively. The soybeans are soaked in the selenium-enriched nutrient solution for 25 hours. After 8 hours of soaking, the soybeans are sonicated under the following conditions: ultrasonic frequency of 45 kHz and ultrasonic intensity density of 0.6 W / cm³. 2The ultrasonic treatment time was 20 minutes; the soybean culture temperature was 27℃ and the culture time was 25 hours. After the culture was completed, the soybeans were washed with tap water and drained. The drained soybeans were then labeled as dry soybeans. The drained soybeans were mixed with 3 times their weight of water and ground into a paste using a grinder. The heating temperature of the soybean paste was controlled, and the heating time was 15 minutes. After cooling to room temperature, the soybean paste was enzymatically hydrolyzed for 45 minutes at its natural pH and 50℃ with the addition of cellulase at a dosage of 25 U / g of dry soybeans. Then, the pH of the soybean paste was adjusted to 9.0 and the temperature was adjusted to 50℃ with NaOH solution. Alkaline protease was added and the enzymatic hydrolysis continued for 3.5 hours with a dosage of 30 U / g of dry soybeans. After the enzymatic hydrolysis was completed, the enzyme was inactivated by heating at 100℃ for 5 minutes. The hydrolysate was centrifuged at 5000g for 10 minutes, and the supernatant was collected and the precipitate was discarded. The supernatant was concentrated and dried to obtain selenium-enriched soybean hydrolysate.

[0093] Figure 13 The effect of cooking temperature of soybean slurry on the total selenium, organic selenium content and organic selenium conversion rate in soybean enzymatic hydrolysate; by Figure 13 It can be seen that as the heating temperature of soybean slurry increases, the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate gradually increase, especially when the heating temperature is 125℃, the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate increase significantly. This is related to the fact that heating causes moderate denaturation of soybeans in the slurry, which is beneficial for subsequent enzymatic hydrolysis and dissolution. However, after the heating temperature of the slurry exceeds 125℃, the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate decrease rapidly. This is related to the excessive denaturation and precipitation of proteins (the main carriers of organic selenium) in the slurry due to excessive heating.

[0094] XIV. Investigation into the enzymatic hydrolysis temperature after cellulase enzyme application

[0095] Wash soybeans with clean water; then soak them in a selenium-enriched nutrient solution. The selenium, vitamin C, vitamin B1, and vitamin B7 content in the selenium-enriched nutrient solution are 30 mg / L, 2000 mg / L, 12.5 mg / L, and 60 μg / L, respectively. The soybeans are soaked in the selenium-enriched nutrient solution for 25 hours. After 8 hours of soaking, the soybeans are sonicated under the following conditions: ultrasonic frequency of 45 kHz and ultrasonic intensity density of 0.6 W / cm³. 2The ultrasonic treatment time was 20 min; the soybean cultivation temperature was 27℃ and the cultivation time was 25 h. After cultivation, the soybeans were washed with tap water and drained. The drained soybeans were then labeled as dry soybeans. The drained soybeans were mixed with 3 times their weight of water and ground into a paste using a grinder. The soybean paste was cooked at 125℃ for 15 min. The soybean paste was then enzymatically hydrolyzed with cellulase at its natural pH for 45 min, with the hydrolysis temperature controlled. The amount of cellulase added was 25 U / g of dry soybeans. Then, the pH of the soybean paste was adjusted to 9.0 and the temperature to 50℃ using NaOH solution. Alkaline protease was added and the hydrolysis continued for 3.5 h. The amount of alkaline protease added was 30 U / g of dry soybeans. After the hydrolysis was completed, the enzyme was inactivated by heating at 100℃ for 5 min. The hydrolysate was centrifuged at 5000g for 10 min. The supernatant was collected, and the precipitate was discarded. The supernatant was concentrated and dried to obtain selenium-enriched soybean hydrolysate.

[0096] Figure 14 The effect of cellulase hydrolysis temperature on the total selenium, organic selenium content, and organic selenium conversion rate of soybean hydrolysates; by Figure 14 It was found that with the increase of enzymatic hydrolysis temperature after the addition of cellulase, the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate gradually increased. In particular, the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate significantly increased at an enzymatic hydrolysis temperature of 50℃. This is related to the optimal enzymatic hydrolysis temperature of cellulase, which is conducive to the degradation of cellulose, thereby releasing proteins containing organic selenium through enzymatic hydrolysis. However, after adding cellulase, the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate decreased rapidly after the enzymatic hydrolysis temperature exceeded 60℃. This is related to the gradual inactivation of cellulose at excessively high temperatures.

[0097] XV. Investigation into the enzymatic hydrolysis time after cellulase enzyme application

[0098] Wash soybeans with clean water; then soak them in a selenium-enriched nutrient solution. The selenium, vitamin C, vitamin B1, and vitamin B7 content in the selenium-enriched nutrient solution are 30 mg / L, 2000 mg / L, 12.5 mg / L, and 60 μg / L, respectively. The soybeans are soaked in the selenium-enriched nutrient solution for 25 hours. After 8 hours of soaking, the soybeans are sonicated under the following conditions: ultrasonic frequency of 45 kHz and ultrasonic intensity density of 0.6 W / cm³. 2The ultrasonic treatment time was 20 minutes; the soybean cultivation temperature was 27℃ and the cultivation time was 25 hours. After cultivation, the soybeans were washed with tap water and drained. The drained soybeans were then labeled as dry soybeans. The drained soybeans were mixed with 3 times their weight of water and ground into a paste using a grinder. The soybean paste was cooked at 125℃ for 15 minutes. The soybean paste was then enzymatically hydrolyzed with cellulase at its natural pH value at a temperature of 50℃, with a cellulase addition of 25 U / g of dry soybeans. The pH value of the soybean paste was then adjusted to 9.0 and the temperature to 50℃ using NaOH solution. Alkaline protease was added and the enzymatic hydrolysis continued for 3.5 hours, with an alkaline protease addition of 30 U / g of dry soybeans. After the enzymatic hydrolysis was completed, the enzyme was inactivated by heating at 100℃ for 5 minutes. The hydrolysate was then centrifuged at 5000g for 10 minutes. The supernatant was collected, and the precipitate was discarded. The supernatant was concentrated and dried to obtain selenium-enriched soybean hydrolysate.

[0099] Figure 15 The effect of cellulase hydrolysis time on the total selenium, organic selenium content, and organic selenium conversion rate of soybean hydrolysate; by Figure 15 It can be seen that with the extension of enzymatic hydrolysis time after the addition of cellulase, the total selenium, organic selenium content and organic selenium conversion rate in soybean enzymatic hydrolysate gradually increase. In particular, when the enzymatic hydrolysis time is increased from 25 min to 45 min, the total selenium, organic selenium content and organic selenium conversion rate in soybean enzymatic hydrolysate increase significantly. After that, the increase in total selenium, organic selenium content and organic selenium conversion rate in soybean enzymatic hydrolysate is not significant. Considering the processing cost and time cost, a cellulase enzymatic hydrolysis time of 30-60 min is more appropriate.

[0100] XVI. Investigation into the Dosage of Cellulase Addition

[0101] Wash soybeans with clean water; then soak them in a selenium-enriched nutrient solution. The selenium, vitamin C, vitamin B1, and vitamin B7 content in the selenium-enriched nutrient solution are 30 mg / L, 2000 mg / L, 12.5 mg / L, and 60 μg / L, respectively. The soybeans are soaked in the selenium-enriched nutrient solution for 25 hours. After 8 hours of soaking, the soybeans are sonicated under the following conditions: ultrasonic frequency of 45 kHz and ultrasonic intensity density of 0.6 W / cm³. 2The ultrasonic treatment time was 20 min; the soybean cultivation temperature was 27℃ and the cultivation time was 25 h. After cultivation, the soybeans were washed with tap water and drained. The drained soybeans were then labeled as dry soybeans. The drained soybeans were mixed with 3 times their weight of water and ground into a paste using a grinder. The soybean paste was cooked at 125℃ for 15 min. Cellulase was added to the soybean paste at its natural pH value, and the amount added was adjusted to control the enzymatic hydrolysis temperature at 50℃ and the enzymatic hydrolysis time at 45 min. Then, the pH value of the soybean paste was adjusted to 9.0 and the temperature to 50℃ using NaOH solution. Alkaline protease was added and enzymatic hydrolysis continued for 3.5 h. The amount of alkaline protease added was 30 U / g of dry soybeans. After enzymatic hydrolysis, the enzyme was inactivated by heating at 100℃ for 5 min. The enzymatic hydrolysate was centrifuged at 5000g for 10 min. The supernatant was collected and the precipitate was discarded. The supernatant was concentrated and dried to obtain selenium-enriched soybean enzymatic hydrolysate.

[0102] Figure 16 The effect of cellulase addition on the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate; by Figure 16 It can be seen that with the increase of the amount of cellulase added, the total selenium, organic selenium content and organic selenium conversion rate in soybean enzymatic hydrolysate gradually increased. In particular, when the amount of cellulase added was 30 U / g, the total selenium, organic selenium content and organic selenium conversion rate in soybean enzymatic hydrolysate increased significantly, reaching the highest level. After that, the increase in the total selenium, organic selenium content and organic selenium conversion rate in soybean enzymatic hydrolysate was not significant. Considering the processing cost and time cost, the amount of cellulase added was more appropriate at 10-40 U / g.

[0103] XVII. Investigation of pH of the enzymatic hydrolysate after alkaline protease digestion

[0104] Wash soybeans with clean water; then soak them in a selenium-enriched nutrient solution. The selenium, vitamin C, vitamin B1, and vitamin B7 content in the selenium-enriched nutrient solution are 30 mg / L, 2000 mg / L, 12.5 mg / L, and 60 μg / L, respectively. The soybeans are soaked in the selenium-enriched nutrient solution for 25 hours. After 8 hours of soaking, the soybeans are sonicated under the following conditions: ultrasonic frequency of 45 kHz and ultrasonic intensity density of 0.6 W / cm³. 2The ultrasonic treatment time was 20 min; the soybean cultivation temperature was 27℃ and the cultivation time was 25 h. After cultivation, the soybeans were washed with tap water and drained. The drained soybeans were then labeled as dry soybeans. The drained soybeans were mixed with 3 times their weight of water and ground into a paste using a grinder. The soybean paste was cooked at 125℃ for 15 min. The soybean paste was then enzymatically hydrolyzed with cellulase at its natural pH value at 50℃ for 45 min. The pH value of the soybean paste was then adjusted with NaOH solution, and the temperature of the soybean paste was controlled at 50℃. Alkaline protease was added and the enzymatic hydrolysis continued for 3.5 h at a dosage of 30 U / g of dry soybeans. After the enzymatic hydrolysis was completed, the enzyme was inactivated by heating at 100℃ for 5 min. The hydrolysate was centrifuged at 5000g for 10 min, and the supernatant was collected and the precipitate was discarded. The supernatant was concentrated and dried to obtain selenium-enriched soybean hydrolysate.

[0105] Figure 17 The effect of pH on the total selenium, organic selenium content, and organic selenium conversion rate of soybean hydrolysate by alkaline protease hydrolysis; Figure 17 It can be seen that as the pH of the enzymatic hydrolysate increases after the addition of alkaline protease, the total selenium, organic selenium content and organic selenium conversion rate in soybean enzymatic hydrolysate gradually increase. In particular, when the pH of the enzymatic hydrolysate is increased to 9, the total selenium, organic selenium content and organic selenium conversion rate in soybean enzymatic hydrolysate increase significantly. After that, the total selenium, organic selenium content and organic selenium conversion rate in soybean enzymatic hydrolysate decrease rapidly. This is related to the fact that alkaline protease has the optimal enzymatic hydrolysis pH.

[0106] 18. Investigation into the enzymatic hydrolysis temperature of alkaline protease

[0107] Wash soybeans with clean water; then soak them in a selenium-enriched nutrient solution. The selenium, vitamin C, vitamin B1, and vitamin B7 content in the selenium-enriched nutrient solution are 30 mg / L, 2000 mg / L, 12.5 mg / L, and 60 μg / L, respectively. The soybeans are soaked in the selenium-enriched nutrient solution for 25 hours. After 8 hours of soaking, the soybeans are sonicated under the following conditions: ultrasonic frequency of 45 kHz and ultrasonic intensity density of 0.6 W / cm³. 2The ultrasonic treatment time was 20 min; the soybean cultivation temperature was 27℃ and the cultivation time was 25 h. After cultivation, the soybeans were washed with tap water and drained. The drained soybeans were then labeled as dry soybeans. The drained soybeans were mixed with 3 times their weight of water and ground into a paste using a grinder. The soybean paste was cooked at 125℃ for 15 min. The soybean paste was then hydrolyzed with cellulase at its natural pH value to pH 9 for 45 min. The pH value of the soybean paste was then adjusted to 9.0 with NaOH solution, the hydrolysis temperature was changed, and alkaline protease was added to continue the hydrolysis for 3.5 h. The amount of alkaline protease added was 30 U / g of dry soybeans. After the hydrolysis was completed, the enzyme was inactivated by heating at 100℃ for 5 min. The hydrolysate was centrifuged at 5000g for 10 min, and the supernatant was collected and the precipitate was discarded. The supernatant was concentrated and dried to obtain selenium-enriched soybean hydrolysate.

[0108] Figure 18 The effect of alkaline protease hydrolysis temperature on the total selenium, organic selenium content, and organic selenium conversion rate of soybean hydrolysate was determined by... Figure 18 It can be seen that with the increase of enzymatic hydrolysis temperature after the addition of alkaline protease, the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate gradually increase, especially when the alkaline protease addition reaches 50℃, the total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate significantly increase. However, when the enzymatic hydrolysis temperature is higher than 50℃, the increase in total selenium, organic selenium content, and organic selenium conversion rate in soybean enzymatic hydrolysate rapidly decreases, which is related to the fact that the high temperature exceeds the optimal enzymatic hydrolysis temperature of alkaline protease.

[0109] 19. Investigation into the enzymatic hydrolysis time after alkaline protease

[0110] Wash soybeans with clean water; then soak them in a selenium-enriched nutrient solution. The selenium, vitamin C, vitamin B1, and vitamin B7 content in the selenium-enriched nutrient solution are 30 mg / L, 2000 mg / L, 12.5 mg / L, and 60 μg / L, respectively. The soybeans are soaked in the selenium-enriched nutrient solution for 25 hours. After 8 hours of soaking, the soybeans are sonicated under the following conditions: ultrasonic frequency of 45 kHz and ultrasonic intensity density of 0.6 W / cm³. 2The ultrasonic treatment time was 20 min; the soybean cultivation temperature was 27℃ and the cultivation time was 25 h. After cultivation, the soybeans were washed with tap water and drained. The drained soybeans were then labeled as dry soybeans. The drained soybeans were mixed with 3 times their weight of water and ground into a paste using a grinder. The soybean paste was cooked at 125℃ for 15 min. Cellulase was added to the soybean paste at its natural pH value to hydrolyze it at pH 9 for 45 min. Then, the pH value of the soybean paste was adjusted to 9.0 with NaOH solution at 50℃, and the hydrolysis time was changed. The amount of alkaline protease added was 30 U / g of dry soybeans. After hydrolysis, the enzyme was inactivated by heating at 100℃ for 5 min. The hydrolysate was centrifuged at 5000g for 10 min, and the supernatant was collected and the precipitate was discarded. The supernatant was concentrated and dried to obtain selenium-enriched soybean hydrolysate.

[0111] Figure 19 The effect of alkaline protease hydrolysis time on the total selenium, organic selenium content, and organic selenium conversion rate of soybean hydrolysate; by Figure 19 It can be seen that with the extension of the hydrolysis time after adding alkaline protease, the total selenium, organic selenium content and organic selenium conversion rate in soybean hydrolysate gradually increase, especially when the hydrolysis time reaches 3.5 hours, the total selenium, organic selenium content and organic selenium conversion rate in soybean hydrolysate increase significantly; after continuing to increase the hydrolysis time to 6 hours, the total selenium, organic selenium content and organic selenium conversion rate in soybean hydrolysate basically no longer increase; taking all factors into consideration, a hydrolysis time of 1-6 hours is appropriate.

[0112] 20. Investigation into the amount of alkaline protease added

[0113] Wash soybeans with clean water; then soak them in a selenium-enriched nutrient solution. The selenium, vitamin C, vitamin B1, and vitamin B7 content in the selenium-enriched nutrient solution are 30 mg / L, 2000 mg / L, 12.5 mg / L, and 60 μg / L, respectively. The soybeans are soaked in the selenium-enriched nutrient solution for 25 hours. After 8 hours of soaking, the soybeans are sonicated under the following conditions: ultrasonic frequency of 45 kHz and ultrasonic intensity density of 0.6 W / cm³. 2The ultrasonic treatment time was 20 min; the soybean cultivation temperature was 27℃ and the cultivation time was 25 h. After cultivation, the soybeans were washed with tap water and drained. The drained soybeans were then labeled as dry soybeans. The drained soybeans were mixed with 3 times their weight of water and ground into a paste using a grinder. The soybean paste was cooked at 125℃ for 15 min. The soybean paste was then hydrolyzed with cellulase at its natural pH value until pH 9 was reached, and the hydrolysis time was 45 min. Then, the pH value of the soybean paste was adjusted to 9.0 with NaOH solution, and the temperature was 50℃. The amount of alkaline protease added was varied for 3.5 h of hydrolysis. After hydrolysis, the enzyme was inactivated by heating at 100℃ for 5 min. The hydrolysate was centrifuged at 5000g for 10 min, and the supernatant was collected and the precipitate was discarded. The supernatant was concentrated and dried to obtain selenium-enriched soybean hydrolysate.

[0114] Figure 20 The effect of alkaline protease addition on total selenium, organic selenium content, and organic selenium conversion rate of soybean enzymatic hydrolysate; by Figure 20 It can be seen that with the increase of alkaline protease addition, the total selenium, organic selenium content and organic selenium conversion rate in soybean enzymatic hydrolysate gradually increase, especially when the alkaline protease addition is 30 U / g, the total selenium, organic selenium content and organic selenium conversion rate in soybean enzymatic hydrolysate increase significantly; thereafter, with the increase of alkaline protease addition, the increase of total selenium, organic selenium content and organic selenium conversion rate in soybean enzymatic hydrolysate is not significant. Considering the processing cost and time cost, the alkaline protease addition is more appropriate at 10-40 U / g.

[0115] 21. Functional Study of Selenium-Enriched Soybean Enzymatic Hydrolysates

[0116] Volunteers with hyperlipidemia and hyperglycemia (aged 35-70 years) were randomly divided into a control group and an experimental group (as shown in Table 2, 30 people in each group, half male and half female). Control group 1 followed the normal diet, control group 2 each person took 0.8g / kg BW of non-selenium-enriched soybean enzymatic hydrolysate daily, and the experimental group took selenium-enriched soybean enzymatic hydrolysate, the dosage of which is shown in Table 2. After 20 days of following the above diet, fasting blood samples were collected to measure the various indicators in Table 2.

[0117] As shown in Table 2, with the increase of the intake of selenium-enriched soybean enzymatic hydrolysate (indicators marked with S), the levels of triglycerides, total cholesterol, low-density lipoprotein, and blood glucose in patients with hyperlipidemia and hyperglycemia gradually decreased. Compared with controls 1 and 2, when the daily intake of selenium-enriched soybean enzymatic hydrolysate (indicators marked with S) in patients with hyperlipidemia and hyperglycemia exceeded 0.2-0.4 g / kg·BW, the levels of triglycerides, total cholesterol, low-density lipoprotein, and blood glucose decreased significantly (p<0.05), indicating that the selenium-enriched soybean enzymatic hydrolysate produced by the method of this invention has the ability to significantly reduce the above indicators. Meanwhile, in patients with hyperlipidemia and hyperglycemia, the high-density lipoprotein (HDL) level gradually increased with the increase in the intake of selenium-enriched soybean enzymatic hydrolysate. Compared with controls 1 and 2, when the daily intake of selenium-enriched soybean enzymatic hydrolysate exceeded 0.4 g / kg·BW, the increase in HDL level in patients with hyperlipidemia and hyperglycemia reached a significant level (p<0.05), indicating that the selenium-enriched soybean enzymatic hydrolysate produced by the method of this invention has the ability to significantly increase the HDL level in patients. It is worth noting that the selenium-enriched soybean residue enzymatic hydrolysate produced by the method of this invention also has a certain ability to reduce the levels of triglycerides, total cholesterol, low-density lipoprotein (LDL), and blood glucose in patients, and to increase the level of HDL level in patients. When the intake is greater than 2.0 g / kg·BW, its ability to improve blood lipids and blood glucose is significantly better than that of controls 1 and 2 (p<0.05). However, its effect is still significantly lower than that of selenium-enriched soybean enzymatic hydrolysate. Moreover, selenium-enriched soybean residue enzymatic hydrolysate has a poor taste, and excessive intake of selenium-enriched soybean residue enzymatic hydrolysate may reduce consumers' quality of life. Therefore, selenium-enriched soybean enzymatic hydrolysate produced by the method of this invention is a better product for lowering blood lipids and blood sugar.

[0118] Table 2. Effects of selenium-enriched soybean hydrolysate and selenium-enriched soybean residue hydrolysate on patients with hyperlipidemia and hyperglycemia.

[0119]

[0120]

[0121] Note: 1. Data in the same column do not contain the same letters, indicating that there is a significant difference between the values ​​(p<0.05); 2. S and C represent patients who take selenium-enriched soybean hydrolysate and patients who take selenium-enriched soybean residue hydrolysate, respectively; (1) Selenium-enriched soybean hydrolysate was prepared under the following conditions: Soybeans were washed with water, and the washed soybeans were soaked in selenium-enriched nutrient solution for 25 hours. The selenium content in the selenium-enriched nutrient solution was 30 mg / L, and the selenium was derived from sodium selenite. The contents of vitamin C, vitamin B1 and vitamin B7 in the selenium-enriched nutrient solution were 2000 mg / L, 12.5 mg / L and 60 μg / L, respectively. Ultrasound was performed on the 8th hour after the soybeans were soaked to promote the absorption and conversion of selenium and other nutrients. The ultrasound conditions were a frequency of 45 kHz and an intensity density of 0.6 W / cm. 21. Time: 2 minutes; 2. After cultivation, drain; 3. After draining, cultivate at 27℃ for 25 hours. After cultivation, wash with tap water and drain again. The drained soybeans are then labeled as dry soybeans. 4. Mix the drained soybeans with 3 times their weight of water and grind them using a grinder to separate the pulp from the residue, collecting the soybean slurry. 5. Heat the soybean slurry at 125℃ for 15 minutes, and then quickly cool it to room temperature. 6. Add cellulase to the soybean slurry at its natural pH and 50℃ for 45 minutes (25 U / g dry soybeans). 7. Adjust the pH to 90.0 and temperature to 50℃ with NaOH solution, and add alkaline protease for 3.5 hours of enzymatic hydrolysis (30 U / g dry soybeans). After the end, heat at 100℃ for 5 min to inactivate the enzyme. Centrifuge the enzymatic hydrolysate at 5000g for 10 min and take the supernatant. After concentration and drying, the supernatant is used to obtain selenium-enriched soybean enzymatic hydrolysate. (2) Selenium-enriched soybean residue enzymatic hydrolysate was prepared under the following conditions: Soybeans were washed with water and the cleaned soybeans were soaked in selenium-enriched nutrient solution for 25 h. The selenium content in the selenium-enriched nutrient solution was 30 mg / L, and the selenium was derived from sodium selenite. The contents of vitamin C, vitamin B1 and vitamin B7 in the selenium-enriched nutrient solution were 2000 mg / L, 12.5 mg / L and 60 μg / L, respectively. Ultrasound was performed on the 8th hour after the soybeans were soaked to promote the absorption and conversion of selenium and other nutrients. The ultrasound conditions were a frequency of 45 kHz and an intensity density of 0.6 W / cm. 2 The soybeans were cultured for 2 minutes; after cultivation, they were drained. The drained soybeans were then cultured at 27℃ for 25 hours. After cultivation, they were washed with tap water and drained. The drained soybeans were then recorded as dry soybeans. The drained soybeans were mixed with 3 times their weight of water and ground with a grinder to separate the pulp from the residue. The soybean residue was then mixed with 2 times its weight of water and heated at 125℃ for 15 minutes. After cooking, the mixture was quickly cooled to room temperature. The cooked soybean residue mixture was then enzymatically hydrolyzed for 45 minutes at its natural pH and 50℃ with the addition of cellulase at a dosage of 25 U / g of dry soybeans. The pH of the mixture was then adjusted to 90.0 and the temperature to 50℃ with NaOH solution. Alkaline protease was added and the enzymatic hydrolysis continued for 3.5 hours at a dosage of 30 U / g of dry soybeans. After the enzymatic hydrolysis, the mixture was heated at 100℃ for 5 minutes to inactivate the enzyme. The hydrolysate was then concentrated and dried to obtain the control. 3. Control 1 was the patient in the routine diet group, and control 2 was the patient who took non-selenium-enriched soybean hydrolysate. The preparation method was the same as (1), except that the patient was not soaked in selenium-enriched nutrient solution (soaked in water), and the intake was 0.8g / kg·BW.

[0122] Example 1:

[0123] Soybeans were rinsed with clean water and then soaked in a selenium-enriched nutrient solution for 10 hours. The selenium content in the solution was 12 mg / L, derived from sodium selenite. The solution also contained 500 mg / L of vitamin C, 2 mg / L of vitamin B1, and 10 μg / L of vitamin B7. After soaking for 8 hours, the soybeans were sonicated to promote the absorption and conversion of selenium and other nutrients. The sonication conditions were 20 kHz frequency and 0.1 W / cm² intensity. 2 1. Incubation time: 3 minutes; 2. Drain the soybeans after incubation; 3. Incubate the drained soybeans at 20℃ for 4 hours, then rinse with tap water and drain again; 4. The drained soybeans are now called dry soybeans; 5. Mix the drained soybeans with twice their weight of water and grind them using a grinder to separate the pulp from the residue, collecting the soybean slurry; 6. Heat the soybean slurry at 100℃ for 10 seconds, then quickly cool it to room temperature; 7. Add cellulase to the soybean slurry at its natural pH and 30℃ for 30 minutes, with a cellulase addition rate of 10 U / g dry soybeans. Then, the pH of the soybean slurry was adjusted to 8.0 and the temperature to 35℃ using NaOH solution. Alkaline protease was added and enzymatic hydrolysis continued for 1 hour. The amount of alkaline protease added was 10 U / g of dry soybeans. After the enzymatic hydrolysis was completed, the enzyme was inactivated by heating at 100℃ for 5 minutes. The hydrolysate was centrifuged at 5000g for 10 minutes, and the supernatant was collected. The supernatant was concentrated and dried to obtain selenium-enriched soybean hydrolysate. The total selenium content in the selenium-enriched soybean hydrolysate was 5.98 mg / kg, and the proportion of organic selenium was 95.12%.

[0124] Volunteers with hyperlipidemia and hyperglycemia (aged 35-70 years) were randomly divided into a control group and an experimental group (as shown in Table 3, 30 people in each group, half male and half female). Control group 1 followed the normal diet, control group 2 each person took 0.8g / kg BW of non-selenium-enriched soybean enzymatic hydrolysate daily, and the experimental group took selenium-enriched soybean enzymatic hydrolysate, the dosage of which is shown in Table 3. After taking the above method for 20 days, fasting blood samples were collected to measure the various indicators in Table 3.

[0125] As shown in Table 3, with the increase in the amount of selenium-enriched soybean hydrolysate produced by the method of this invention in patients with hyperglycemia and hyperlipidemia, the levels of triglycerides, total cholesterol, low-density lipoprotein (LDL), and blood glucose gradually decreased, while the level of high-density lipoprotein (HDL) gradually increased. Compared with Control 1 and Control 2, when the daily intake of selenium-enriched soybean hydrolysate reached 0.8 g / kg·BW, the decrease in the levels of triglycerides, total cholesterol, LDL, and blood glucose was significant (p<0.05), and the increase in the level of HDL was also significant (p<0.05).

[0126] As patients with hyperglycemia and hyperlipidemia increased their intake of the selenium-enriched soybean residue enzymatic hydrolysate produced by the method of this invention, their triglyceride, total cholesterol, low-density lipoprotein (LDL), and blood glucose levels gradually decreased, while their high-density lipoprotein (HDL) level gradually increased. Compared with controls 1 and 2, when patients' daily intake of selenium-enriched soybean residue enzymatic hydrolysate reached 2.5 g / kg·BW, the decrease in triglyceride, total cholesterol, LDL, and blood glucose levels was significant (p<0.05). When patients' daily intake of selenium-enriched soybean residue enzymatic hydrolysate reached 1.2 g / kg·BW, the increase in HDL levels was significant (p<0.05). However, compared with selenium-enriched soybean residue enzymatic hydrolysate, the lipid-lowering and blood glucose-lowering effects of selenium-enriched soybean residue enzymatic hydrolysate were poor. To achieve the same effect as selenium-enriched soybean residue enzymatic hydrolysate, a large amount of selenium-enriched soybean residue enzymatic hydrolysate would be required. Because selenium-enriched soybean residue has a poor taste, the selenium-enriched soybean residue enzymatic hydrolysate produced by this invention has a greater advantage.

[0127] Table 3. Effects of selenium-enriched soybean hydrolysate and selenium-enriched soybean residue hydrolysate on patients with hyperlipidemia and hyperglycemia.

[0128]

[0129]

[0130] Note: 1. The absence of identical letters in the same column indicates a significant difference between the numbers (p<0.05); 2. S and C represent patients who took selenium-enriched soybean hydrolysate and patients who took selenium-enriched soybean residue hydrolysate, respectively; 3. The preparation methods of selenium-enriched soybean residue hydrolysate, control 1, and control 2 are the same as those recorded in the corresponding contents of Table 2.

[0131] Example 2:

[0132] Soybeans were rinsed with clean water and then soaked in a selenium-enriched nutrient solution for 15 hours. The selenium content in the solution was 24 mg / L, derived from sodium selenate. The solution also contained 1625 mg / L of vitamin C, 6.5 mg / L of vitamin B1, and 32.5 μg / L of vitamin B7. After soaking for 8 hours, the soybeans were sonicated to promote the absorption and conversion of selenium and other nutrients. The sonication conditions were 32 kHz frequency and 0.45 W / cm² intensity. 2The process involves: 1. Incubation time: 9.8 min; 2. Draining: The drained soybeans are incubated at 23℃ for 13 h, then rinsed with tap water and drained again. The drained soybeans are then labeled as dry soybeans. The drained soybeans are mixed with three times their weight of water and ground using a grinder to separate the pulp from the residue, yielding the soybean slurry. The soybean slurry is heated at 109℃ for 15 min, then rapidly cooled to room temperature. The soybean slurry is then enzymatically hydrolyzed with cellulase at its natural pH and 37.5℃ for 37.5 min, with a cellulase addition amount of 1... 7.5 U / g of dried soybeans were used, and the pH of the soybean slurry was adjusted to 8.5 and the temperature to 40℃ using NaOH solution. Alkaline protease was added and enzymatic hydrolysis was continued for 2.25 h. The amount of alkaline protease added was 17.5 U / g of dried soybeans. After the enzymatic hydrolysis was completed, the enzyme was inactivated by heating at 100℃ for 5 min. The hydrolysate was centrifuged at 5000g for 10 min, and the supernatant was collected. The supernatant was concentrated and dried to obtain selenium-enriched soybean hydrolysate. The total selenium content of the selenium-enriched soybean hydrolysate was 12.18 mg / kg, and the proportion of organic selenium was 96.26%.

[0133] Volunteers with hyperlipidemia and hyperglycemia (aged 38-75 years) were randomly divided into a control group and an experimental group (as shown in Table 4, 30 people in each group, half male and half female). Control group 1 followed the normal diet, control group 2 each took 0.8 g / kg BW of non-selenium-enriched soybean enzymatic hydrolysate daily, and the experimental group took selenium-enriched soybean enzymatic hydrolysate, the dosage of which is shown in Table 4. After taking the above method for 20 days, fasting blood samples were collected to measure the various indicators in Table 4.

[0134] As shown in Table 4, with the increase in the amount of selenium-enriched soybean hydrolysate produced by the method of this invention in patients with hyperglycemia and hyperlipidemia, the levels of triglycerides, total cholesterol, low-density lipoprotein (LDL), and blood glucose gradually decreased, while the level of high-density lipoprotein (HDL) gradually increased. Compared with Control 1 and Control 2, when the daily intake of selenium-enriched soybean hydrolysate reached 0.8 g / kg·BW, the decrease in the levels of triglycerides, total cholesterol, LDL, and blood glucose was significant (p<0.05). When the daily intake of selenium-enriched soybean hydrolysate reached 0.4 g / kg·BW, the increase in the level of HDL was significant (p<0.05).

[0135] As patients with hyperglycemia and hyperlipidemia increased their intake of the selenium-enriched soybean residue enzymatic hydrolysate produced by the method of this invention, their triglyceride, total cholesterol, low-density lipoprotein (LDL), and blood glucose levels gradually decreased, while their high-density lipoprotein (HDL) level gradually increased. Compared with controls 1 and 2, when patients' daily intake of selenium-enriched soybean residue enzymatic hydrolysate reached 2.5 g / kg·BW, the decrease in triglyceride, total cholesterol, LDL, and blood glucose levels was significant (p<0.05). When patients' daily intake of selenium-enriched soybean residue enzymatic hydrolysate reached 1.2 g / kg·BW, the increase in HDL levels was significant (p<0.05). However, compared with selenium-enriched soybean residue enzymatic hydrolysate, the overall effect of selenium-enriched soybean residue enzymatic hydrolysate in lowering blood lipids and blood glucose was poorer. To achieve the same effect as selenium-enriched soybean residue enzymatic hydrolysate, a large amount of selenium-enriched soybean residue enzymatic hydrolysate would be required. Because selenium-enriched soybean residue has a poor taste, the selenium-enriched soybean residue enzymatic hydrolysate produced by this invention has a greater advantage.

[0136] Table 4. Effects of selenium-enriched soybean hydrolysate and selenium-enriched soybean residue hydrolysate on patients with hyperlipidemia and hyperglycemia.

[0137]

[0138]

[0139] Note: 1. The absence of identical letters in the same column indicates a significant difference between the numbers (p<0.05); 2. S and C represent patients who took selenium-enriched soybean hydrolysate and patients who took selenium-enriched soybean residue hydrolysate, respectively; 3. The preparation methods of selenium-enriched soybean residue hydrolysate, control 1, and control 2 are the same as those recorded in the corresponding contents of Table 2.

[0140] Example 3:

[0141] Soybeans were rinsed with clean water and then soaked in a selenium-enriched nutrient solution for 20 hours. The selenium content in the solution was 36 mg / L, derived from nano-selenium. The solution also contained 2750 mg / L of vitamin C, 11 mg / L of vitamin B1, and 55 μg / L of vitamin B7. After soaking for 8 hours, the soybeans were sonicated to promote the absorption and conversion of selenium and other nutrients. The sonication conditions were 42 kHz frequency and 0.80 W / cm² intensity. 2The incubation time was 16.6 min; after incubation, the soybeans were drained; the drained soybeans were incubated at 26℃ for 22 h, and after incubation, they were washed with tap water and drained; the drained soybeans at this time were recorded as dry soybeans; the drained soybeans were mixed with 4 times their weight of water and ground with a grinder to separate the pulp and residue, and the soybean slurry was collected; the soybean slurry was heated at 118℃ for 30 min, and after cooking, it was quickly cooled to room temperature; the soybean slurry was enzymatically hydrolyzed with cellulase at natural pH and 45℃ for 45 min, and the amount of cellulase added was 25 U / g dry soybeans. Soybeans were used, and the pH of the soybean slurry was adjusted to 9 and the temperature to 47℃ using NaOH solution. Alkaline protease was added and enzymatic hydrolysis was continued for 3.5 hours. The amount of alkaline protease added was 25 U / g of dry soybeans. After the enzymatic hydrolysis was completed, the enzyme was inactivated by heating at 100℃ for 5 minutes. The hydrolysate was centrifuged at 5000g for 10 minutes, and the supernatant was collected. The supernatant was concentrated and dried to obtain selenium-enriched soybean hydrolysate. The total selenium content of the selenium-enriched soybean hydrolysate was 18.30 mg / kg, and the proportion of organic selenium was 97.40%.

[0142] Volunteers with hyperlipidemia and hyperglycemia (aged 40-70 years) were randomly divided into a control group and an experimental group (as shown in Table 5, 30 people in each group, half male and half female). Control group 1 followed the normal diet, control group 2 each person took 0.8g / kg BW of non-selenium-enriched soybean enzymatic hydrolysate daily, and the experimental group took selenium-enriched soybean enzymatic hydrolysate, the dosage of which is shown in Table 5. After taking the above method for 20 days, fasting blood samples were collected to measure the various indicators in Table 5.

[0143] As shown in Table 5, with the increase in the amount of selenium-enriched soybean hydrolysate produced by the method of this invention in patients with hyperglycemia and hyperlipidemia, the levels of triglycerides, total cholesterol, low-density lipoprotein (LDL), and blood glucose gradually decreased, while the level of high-density lipoprotein (HDL) gradually increased. Compared with Control 1 and Control 2, when the daily intake of selenium-enriched soybean hydrolysate reached 0.4 g / kg BW, the decrease in the levels of triglycerides, total cholesterol, LDL, and blood glucose was significant (p<0.05). When the daily intake of selenium-enriched soybean hydrolysate reached 0.2 g / kg BW, the increase in the level of HDL was significant (p<0.05).

[0144] As patients with hyperglycemia and hyperlipidemia increased their intake of the selenium-enriched soybean residue enzymatic hydrolysate produced by the method of this invention, their triglyceride, total cholesterol, low-density lipoprotein (LDL), and blood glucose levels gradually decreased, while their high-density lipoprotein (HDL) level gradually increased. Compared with controls 1 and 2, when patients' daily intake of selenium-enriched soybean residue enzymatic hydrolysate reached 2.0 g / kg·BW, the decrease in triglyceride, total cholesterol, LDL, and blood glucose levels was significant (p<0.05). When patients' daily intake of selenium-enriched soybean residue enzymatic hydrolysate reached 0.8 g / kg·BW, the increase in HDL levels was significant (p<0.05). However, compared with selenium-enriched soybean residue enzymatic hydrolysate, the overall effect of selenium-enriched soybean residue enzymatic hydrolysate in lowering blood lipids and blood glucose was poorer. To achieve the same effect as selenium-enriched soybean residue enzymatic hydrolysate, a large amount of selenium-enriched soybean residue enzymatic hydrolysate would be required. Because selenium-enriched soybean residue has a poor taste, the selenium-enriched soybean residue enzymatic hydrolysate produced by this invention has a greater advantage.

[0145] Table 5. Effects of selenium-enriched soybean hydrolysate and selenium-enriched soybean residue hydrolysate on patients with hyperlipidemia and hyperglycemia.

[0146]

[0147]

[0148] Note: 1. Data in the same column that do not contain the same letters indicate a significant difference between the numbers (p<0.05); 2. S and C represent patients who took selenium-enriched soybean hydrolysate and patients who took selenium-enriched soybean residue hydrolysate, respectively.

[0149] Example 4:

[0150] Soybeans were rinsed with clean water and then soaked in a selenium-enriched nutrient solution for 25 hours. The selenium content in the solution was 48 mg / L, derived from nano-selenium. The vitamin C, vitamin B1, and vitamin B7 content in the solution were 3875 mg / L, 15.5 mg / L, and 77.5 μg / L, respectively. After soaking for 8 hours, the soybeans were sonicated to promote the absorption and conversion of selenium and other nutrients. The sonication conditions were a frequency of 54 kHz and an intensity density of 1.15 W / cm². 2The incubation period was 23.3 min; after incubation, the soybeans were drained; the drained soybeans were incubated at 29℃ for 31 h, and after incubation, they were washed with tap water and drained; the drained soybeans at this time were recorded as dry soybeans; the drained soybeans were mixed with 5 times their weight of water and ground with a grinder to separate the pulp and residue, and the soybean slurry was collected; the soybean slurry was heated at 126℃ for 45 min, and after cooking, it was quickly cooled to room temperature; the soybean slurry was enzymatically hydrolyzed with cellulase at natural pH and 52.5℃ for 52.5 min, and the amount of cellulase added was 33 U / g. Dried soybeans were used, and the pH of the soybean slurry was adjusted to 9.5 and the temperature to 53℃ using NaOH solution. Alkaline protease was added and enzymatic hydrolysis was continued for 4.8 hours. The amount of alkaline protease added was 33 U / g of dried soybeans. After the enzymatic hydrolysis was completed, the enzyme was inactivated by heating at 100℃ for 5 minutes. The hydrolysate was centrifuged at 5000g for 10 minutes, and the supernatant was collected. The supernatant was concentrated and dried to obtain selenium-enriched soybean hydrolysate. The total selenium content of the selenium-enriched soybean hydrolysate was 30.13 mg / kg, and the proportion of organic selenium was 99.63%.

[0151] Volunteers with hyperlipidemia and hyperglycemia (aged 45-65 years) were randomly divided into a control group and an experimental group (as shown in Table 6, 30 people in each group, half male and half female). Control group 1 followed the normal diet, control group 2 each person took 0.8g / kg BW of non-selenium-enriched soybean enzymatic hydrolysate daily, and the experimental group took selenium-enriched soybean enzymatic hydrolysate, the dosage of which is shown in Table 6. After taking the above method for 20 days, fasting blood samples were collected to measure the various indicators in Table 6.

[0152] As shown in Table 6, with the increase in the amount of selenium-enriched soybean hydrolysate produced by the method of this invention in patients with hyperglycemia and hyperlipidemia, the levels of triglycerides, total cholesterol, low-density lipoprotein (LDL), and blood glucose gradually decreased, while the level of high-density lipoprotein (HDL) gradually increased. Compared with Control 1 and Control 2, when the daily intake of selenium-enriched soybean hydrolysate reached 0.2 g / kg·BW, the decrease in the levels of triglycerides, total cholesterol, LDL, and blood glucose was significant (p<0.05), and the increase in the level of HDL was significant (p<0.05).

[0153] As patients with hyperglycemia and hyperlipidemia increased their intake of the selenium-enriched soybean residue enzymatic hydrolysate produced by the method of this invention, their triglyceride, total cholesterol, low-density lipoprotein (LDL), and blood glucose levels gradually decreased, while their high-density lipoprotein (HDL) level gradually increased. Compared with controls 1 and 2, when patients' daily intake of selenium-enriched soybean residue enzymatic hydrolysate reached 0.8 g / kg·BW, the decrease in triglyceride, total cholesterol, LDL, and blood glucose levels was significant (p<0.05). When patients' daily intake of selenium-enriched soybean residue enzymatic hydrolysate reached 0.2 g / kg·BW, the increase in HDL levels was significant (p<0.05). However, compared with selenium-enriched soybean residue enzymatic hydrolysate, the overall effect of selenium-enriched soybean residue enzymatic hydrolysate in lowering blood lipids and blood glucose was poorer. To achieve the same effect as selenium-enriched soybean residue enzymatic hydrolysate, a large amount of selenium-enriched soybean residue enzymatic hydrolysate would be required. Because selenium-enriched soybean residue has a poor taste, the selenium-enriched soybean residue enzymatic hydrolysate produced by this invention has a greater advantage.

[0154] Table 6. Effects of selenium-enriched soybean enzymatic hydrolysate and selenium-enriched soybean residue enzymatic hydrolysate on patients with hyperlipidemia and hyperglycemia.

[0155]

[0156]

[0157] Note: 1. The absence of identical letters in the same column indicates a significant difference between the numbers (p<0.05); 2. S and C represent patients who took selenium-enriched soybean hydrolysate and patients who took selenium-enriched soybean residue hydrolysate, respectively; 3. The preparation methods of selenium-enriched soybean residue hydrolysate, control 1, and control 2 are the same as those recorded in the corresponding contents of Table 2.

[0158] Example 5:

[0159] Soybeans were rinsed with clean water and then soaked in a selenium-enriched nutrient solution for 30 hours. The selenium content in the solution was 60 mg / L, derived from nano-selenium. The solution also contained 5000 mg / L of vitamin C, 20 mg / L of vitamin B1, and 100 μg / L of vitamin B7. After soaking for 8 hours, the soybeans were sonicated to promote the absorption and conversion of selenium and other nutrients. The sonication conditions were 65 kHz frequency and 1.5 W / cm² intensity. 21. Incubation time: 30 min; 2. Drain the soybeans after incubation; 3. Incubate the drained soybeans at 32℃ for 40 h, then rinse with tap water and drain again; 4. The drained soybeans are now called dry soybeans; 5. Mix the drained soybeans with 6 times their weight of water and grind them using a grinder to separate the pulp from the residue, collecting the soybean slurry; 6. Heat the soybean slurry at 135℃ for 60 min, then quickly cool it to room temperature after cooking; 7. Add cellulase to the soybean slurry at its natural pH and 60℃ for 60 min, with a cellulase addition rate of 40 U / g dry soybeans. Soybeans were processed, and the pH of the soybean slurry was adjusted to 10 and the temperature to 60℃ using NaOH solution. Alkaline protease was added and enzymatic hydrolysis continued for 6 hours. The amount of alkaline protease added was 40 U / g of dry soybeans. After the enzymatic hydrolysis was completed, the enzyme was inactivated by heating at 100℃ for 5 minutes. The hydrolysate was centrifuged at 5000g for 10 minutes, and the supernatant was collected. The supernatant was concentrated and dried to obtain selenium-enriched soybean hydrolysate. The total selenium content of the selenium-enriched soybean hydrolysate was 27.32 mg / kg, and the proportion of organic selenium was 98.52%.

[0160] Volunteers with hyperlipidemia and hyperglycemia (aged 50-70 years) were randomly divided into a control group and an experimental group (as shown in Table 7, 30 people in each group, half male and half female). Control group 1 followed the normal diet, control group 2 each person took 0.8g / kg BW of non-selenium-enriched soybean enzymatic hydrolysate daily, and the experimental group took selenium-enriched soybean enzymatic hydrolysate, the dosage of which is shown in Table 7. After taking the above method for 20 days, fasting blood samples were collected to measure the various indicators in Table 7.

[0161] As shown in Table 7, with the increase in the amount of selenium-enriched soybean hydrolysate produced by the method of this invention in patients with hyperglycemia and hyperlipidemia, the levels of triglycerides, total cholesterol, low-density lipoprotein (LDL), and blood glucose gradually decreased, while the level of high-density lipoprotein (HDL) gradually increased. Compared with Control 1 and Control 2, when the daily intake of selenium-enriched soybean hydrolysate reached 0.4 g / kg·BW, the decrease in the levels of triglycerides, total cholesterol, LDL, and blood glucose was significant (p<0.05). When the daily intake of selenium-enriched soybean hydrolysate reached 0.2 g / kg·BW, the increase in the level of HDL was significant (p<0.05).

[0162] As patients with hyperglycemia and hyperlipidemia increased their intake of the selenium-enriched soybean residue enzymatic hydrolysate produced by the method of this invention, their triglyceride, total cholesterol, low-density lipoprotein (LDL), and blood glucose levels gradually decreased, while their high-density lipoprotein (HDL) level gradually increased. Compared with controls 1 and 2, when the daily intake of selenium-enriched soybean residue enzymatic hydrolysate reached 1.2 g / kg·BW, the decrease in triglyceride, total cholesterol, LDL, and blood glucose levels was significant (p<0.05). When the daily intake of selenium-enriched soybean residue enzymatic hydrolysate reached 0.4 g / kg·BW, the increase in HDL levels was significant (p<0.05). However, compared with selenium-enriched soybean residue enzymatic hydrolysate, the overall effect of selenium-enriched soybean residue enzymatic hydrolysate in lowering blood lipids and blood glucose was poorer. To achieve the same effect as selenium-enriched soybean residue enzymatic hydrolysate, a large amount of selenium-enriched soybean residue enzymatic hydrolysate would be required. Because selenium-enriched soybean residue has a poor taste, the selenium-enriched soybean residue enzymatic hydrolysate produced by this invention has a greater advantage.

[0163] Table 7. Effects of selenium-enriched soybean hydrolysate and selenium-enriched soybean residue hydrolysate on patients with hyperlipidemia and hyperglycemia.

[0164]

[0165]

[0166] Note: 1. The absence of identical letters in the same column indicates a significant difference between the numbers (p<0.05); 2. S and C represent patients who took selenium-enriched soybean hydrolysate and patients who took selenium-enriched soybean residue hydrolysate, respectively; 3. The preparation methods of selenium-enriched soybean residue hydrolysate, control 1, and control 2 are the same as those recorded in the corresponding contents of Table 2.

[0167] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope defined in the claims.

Claims

1. A method for preparing a selenium-enriched soybean enzymatic hydrolysate having a blood lipid-lowering and blood glucose-lowering function, characterized in that, The steps are as follows: (1) wash the soybeans with clean water and wait for use; (2) soaking the soybeans in a selenium-rich nutrient solution for 10-30 hours, applying ultrasonic treatment during the soaking until the end of the soaking, and draining the water after the end of the soaking to obtain ultrasonically soaked soybeans; the selenium-rich nutrient solution is composed of selenium, vitamin C, vitamin B1, vitamin B7, and water, wherein the content of selenium is 12-60 mg / L, the selenium is derived from sodium selenite, sodium selenate, and nano selenium; the contents of vitamin C, vitamin B1, and vitamin B7 are 500-5000 mg / L, 2-20 mg / L, and 10-100 µg / L, respectively; the ultrasonic treatment conditions are: an ultrasonic frequency of 20-65 kHz, an ultrasonic intensity density of 0.1-1.5 W / cm 2 , and an ultrasonic time of 3-30 min; (3) drying the ultrasonically soaked soybeans to obtain dried ultrasonically soaked soybeans; the drying is performed at a temperature of 40-60°C for 10-30 hours; (3) culture the soybeans after ultrasonic soaking in step (2) at 20-32℃ for 4-40 h, and then wash with tap water and drain; (4) mix the drained soybeans in step (3) with water, grind the mixture, separate the pulp, and collect the soybean slurry; (5) cook the soybean slurry at 100-135℃ for 10 s-60 min, and then cool to room temperature to obtain cooked soybean slurry; (6) add cellulase to the cooked soybean slurry obtained in step (5) at natural pH and 30-60℃, and carry out enzymatic hydrolysis for 30-60 min, wherein the cellulase is added at an amount of 10-40 U / g of dry soybeans; then adjust the pH of the soybean slurry to 8.0-10.0 with NaOH solution; then adjust the temperature to 35-60℃, and add alkaline protease to continue the enzymatic hydrolysis for 1-6 h, wherein the alkaline protease is added at an amount of 10-40 U / g of dry soybeans; after the enzymatic hydrolysis, heat the enzyme to inactivate it, centrifuge the enzyme hydrolysate, and collect the supernatant after centrifugation; (7) concentrate and dry the supernatant obtained in step (6) to obtain selenium-rich soybean hydrolysate; the total selenium content of the selenium-rich soybean hydrolysate is 5.98-30.13 mg / kg, and the organic selenium content accounts for 95.12-99.63%.

2. The method of claim 1, wherein the method of preparing the selenium-enriched soybean enzymatic hydrolysate having a blood lipid and blood sugar lowering function is characterized by, In step (2), the selenium content in the selenium-enriched nutrient solution is 20-30 mg / L, and the selenium is derived from sodium selenite or nano-selenium; the contents of vitamin C, vitamin B1, and vitamin B7 are 1500-2500 mg / L, 10-15 mg / L, and 50-70 µg / L, respectively; the ultrasonic treatment during soaking is performed during the 0-8 h soaking period; the ultrasonic treatment conditions are: ultrasonic frequency of 40-60 kHz and ultrasonic intensity density of 0.3-0.8 W / cm³. 2 The ultrasound time is 15-25 minutes.

3. The process for the preparation of selenium enriched soybean enzymatic hydrolysate having hypolipidemic and hypoglycemic functions as claimed in claim 1, wherein, In step (3), the temperature for culturing the soybeans is 25-30℃, and the time is 20-30 h.

4. The method for preparing selenium-enriched soybean enzymatic hydrolysate with lipid-lowering and blood glucose-lowering functions according to claim 1, characterized in that, In step (4), the drained soybeans are mixed with water, and the weight ratio of water to soybeans is 2-6:

1.

5. The method of claim 1, wherein the method of preparing the selenium-enriched soybean enzymatic hydrolysate having a blood lipid and blood sugar lowering function is characterized by, In step (6), the temperature for adding cellulase for enzymatic hydrolysis is 45-55℃, and the time is 40-50 min; the cellulase is added at an amount of 20-30 U / g of dry soybeans based on the weight of dry soybeans; the pH of the soybean slurry is 8.5-9.5, the temperature for adding alkaline protease for continued enzymatic hydrolysis is 45-55℃, and the enzymatic hydrolysis time is 3-4 h; the alkaline protease is added at an amount of 25-35 U / g of dry soybeans based on the weight of dry soybeans; the temperature for heating the enzyme is 100℃, and the heating time is 5-10 min; the centrifugation condition is centrifugation at 5000 g for 10 min.

6. Selenium-rich soybean hydrolysate prepared by the method of any one of claims 1-5.

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