Preparation method and application of hemp seed protein peptide with sugar absorption blocking effect

Abstract

The present invention provides a method for preparing a hemp seed protein peptide having the effect of blocking sugar absorption and its application, belonging to the technical field of functional active peptides, comprising the following steps: crushing the hemp seed and then defatting it through a low-temperature continuous phase change to obtain defatted hemp meal after defatting, the defatted hemp meal being hydrolyzed by polysaccharidase and protease in succession, and then subjected to low-temperature salting out after the hydrolysis is completed, and then dried and sterilized to obtain the hemp seed protein peptide. The hemp seed protein peptide obtained by the technical solution and preparation method of the present invention has the following effects: 1. The hemp seed protein peptide prepared by the present invention can significantly inhibit the activity of α-amylase and α-glycosidase (the maximum inhibition rate can reach 84.2% and 75.1% respectively), thereby blocking the human body's digestion and absorption of carbohydrates such as starch, dextrin, and disaccharides. 2. The results of double-blind clinical tests show that the hemp seed protein peptide prepared by the present invention can effectively reduce the peak blood sugar level and peak area after a meal, and the effective sugar-free rate can reach 30-60%.

Description

Technical Field

[0001] The invention relates to the technical field of functional active peptide products, and in particular to a preparation method and application of hemp seed protein peptide with the function of blocking sugar absorption. Background Art

[0002] With the increasing prevalence of chronic diseases like diabetes and obesity, the importance of controlling blood sugar has become increasingly prominent, and more and more people are attempting to control their blood sugar through sugar-free diets. Broadly speaking, sugar-free diets refer to strictly restricting carbohydrate intake or using amylase and glycosidase inhibitors to block the digestion and absorption of sugars in order to control blood sugar. Furthermore, sugar-free diets can help improve blood lipids and other indicators, thereby reducing the risk of cardiovascular disease. Regarding weight management, blocking sugar absorption can reduce calorie intake and encourage the body to consume fat for energy, making it a popular choice for those seeking to lose weight. Consequently, sugar-free diets have become a highly sought-after dietary trend in today's society.

[0003] As people's demand for healthy diets continues to increase, the food industry is constantly seeking healthier and more effective natural active ingredients. Hemp seed protein peptides are small molecule active peptides produced from hemp seed protein through a series of enzymatic hydrolysis processes. Numerous studies have confirmed that they have various activities such as antioxidant, blood pressure lowering, immune enhancement, and anti-fatigue.

[0004] Publication number CN112410393B, titled "A Hemp Active Peptide and Its Preparation Method and Application," is a Chinese patent that discloses the steps of first mixing and grinding pre-cooled defatted hemp seed meal with dry ice to obtain hemp seed meal powder, then adding water and calcium carbonate to prepare an alkaline suspension, grinding and screening the suspension, and then performing a two-step enzymatic hydrolysis using a mixed protease and a neutral protease. Finally, the enzymes are inactivated, filtered, centrifuged, decolorized, and sterilized to obtain the hemp active peptide. The hemp active peptide prepared by the above scheme has high purity, a full taste, high antioxidant activity, and has antioxidant, blood pressure lowering, anti-fatigue, and exercise tolerance-enhancing effects. However, the above technical scheme and the products existing in the prior art only focus on antioxidant and blood pressure lowering functions. Summary of the Invention

[0005] In view of this, the present invention provides a hemp seed protein peptide with the effect of blocking sugar absorption, a preparation method and application thereof. The hemp seed protein peptide obtained by the technical solution and preparation method of the present invention can significantly inhibit the activity of α-amylase and α-glucosidase, thereby blocking the human body's digestion and absorption of starch and sucrose. Clinical test results show that the hemp seed protein peptide prepared by the present invention can effectively reduce the postprandial blood sugar peak and peak area, and the effective sugar-free rate can reach 30-60%, providing a new direction for the development of hemp seed.

[0006] To achieve the above objectives, the present invention provides a method for preparing hemp seed protein peptides having the effect of blocking sugar absorption, comprising the following steps: crushing the hemp seeds and then defatting them through low-temperature continuous phase change to obtain defatted hemp meal after defatting; hydrolyzing the defatted hemp meal with polysaccharidase and protease in succession; performing low-temperature salting out after the hydrolysis; and obtaining the hemp seed protein peptides through drying and sterilization.

[0007] Optionally, the extraction agent of the low-temperature continuous phase change is No. 15 solvent oil; the mass volume ratio of the hemp seed to the extraction agent is 1:8~15; the low-temperature continuous phase change includes the following conditions: temperature of 20℃~35℃, pressure of 2.5Mpa~4Mpa, flow rate of 50~80L / h, continuous extraction time of 90min~120min, analysis temperature of 70℃~85℃, and analysis pressure of 0.5Mpa~1Mpa.

[0008] Optionally, the polysaccharide enzymatic hydrolysis includes the following steps: defatted hemp meal is mixed with water at a material-liquid ratio of 1:10-30, the mass of the added polysaccharide enzyme is 10-15% of the mass of the substrate, the pH is adjusted to 3.0-5.5, and the hydrolysis time is 1-4 hours; the substrate is defatted hemp meal and water.

[0009] Optionally, the polysaccharidase is one or a combination of two or more of cellulase, pectinase, amylase, glycosidase, hemicellulose, glucanase, and xylanase.

[0010] Optionally, the protease hydrolysis includes the following steps: inactivating the enzyme in the product after the polysaccharide enzymatic hydrolysis, then adding protease in an amount of 2 to 6% of the mass of the product after the polysaccharide enzymatic hydrolysis, adjusting the pH to 6.0 to 8.5 for hydrolysis, and the hydrolysis time is 15 to 35 minutes; the protease is Bacillus licheniformis protease.

[0011] Optionally, the low-temperature salting out is to centrifugally filter the product after protease hydrolysis to obtain a filtrate, cool it to 2-4°C, continuously add saturated ammonium sulfate solution to the filtrate until the protein flocculates, salt out for 30-90 minutes, and collect the precipitate after centrifugation.

[0012] Optionally, the drying is spray drying, comprising the following steps: using a spray dryer for spray drying, with an air inlet temperature of 15~25°C, an air outlet temperature of 30~40°C, a feed rate of 50~150kg / h, an electrostatic generator voltage of 30-55kV, and an atomizer pressure of 15~20MPa.

[0013] Optionally, the sterilization is ultra-high pressure sterilization, comprising the following steps: during ultra-high pressure sterilization, the working pressure is 100-500 MPa, the working temperature is 25-40° C., and the pressure holding time is 20-90 s.

[0014] In order to achieve the above purpose, the present invention also provides a method for preparing a hemp seed protein peptide with a sugar-breaking effect, the hemp seed protein peptide comprising the following short peptide sequences: SEQ ID NO: 1: Met-Thr-Met-Trp, SEQ ID NO: 2: Tyr-Lys-Pro-Val-Tyr, SEQ ID NO: 3: Thr-Pro-Val-Ser-Ile-Leu, SEQ ID NO: 4: Met-Ala-His-Leu-Phe, SEQ ID NO: 5: Met-Val-Ser-Leu-Tyr.

[0015] The above technical solution of the present invention includes at least the following beneficial effects:

[0016] 1. Low-temperature continuous phase change deep degreasing technology is used in the technical solution of the present invention: the raw materials of existing hemp seed protein peptide products are almost all hemp meal after oil extraction, and unprocessed hemp seed cannot be used directly. This is because hemp seed contains 35~45% oil. If it is used directly for protein extraction or direct enzymatic hydrolysis, it will cause the oil content in the peptide product to be too high, affecting the product taste and storage stability. In addition, high oil content in the preparation process will also cause equipment adhesion, blockage and other problems, increasing labor and maintenance costs. In the present invention, low-temperature continuous phase change technology is applied to the hemp seed degreasing process for the first time. After degreasing through this step, the oil content of hemp meal is reduced to 1~3%, and the oil content of the finished hemp seed protein peptide is reduced to 3~5‰

[0017] 2. The technical solution of the present invention uses polysaccharidase and protease for step-by-step hydrolysis: Most of the existing production processes for hemp seed protein peptide products use alkali extraction and acid precipitation or single enzyme hydrolysis to extract hemp protein for subsequent peptide product production. Alkali extraction and acid precipitation use a strong alkaline environment, which easily causes the spatial structure of the protein to change, thereby losing its original biological activity. Enzymatic extraction can maintain the natural structure and biological activity of the protein to the greatest extent, thereby ensuring its medicinal value, but in some cases, the action of the enzyme may be incomplete, resulting in a low extraction rate. Therefore, the present invention introduces a composite polysaccharide hydrolase and a Bacillus licheniformis protein hydrolase into the production process for the first time. The combined action of multiple polysaccharidases can effectively hydrolyze difficult-to-hydrolyze polysaccharides such as cellulose and pectin in hemp seed cells, destroy the cell wall to promote protein release; and the protease can efficiently perform preliminary degradation of some macromolecular proteins that have not been completely released into the solution, further improving the protein extraction rate.

[0018] 3. Low-temperature salting out is used in the technical solution of the present invention: Most of the existing hemp seed protein peptide preparation processes obtain hemp protein by acid precipitation, but acid precipitation will cause some hemp protein to denature. In addition, direct discharge of the waste liquid after acid precipitation will cause pollution, and the treatment and discharge will increase production costs. The present invention uses low-temperature salting out to obtain hemp protein, which not only ensures the natural activity of hemp protein, but also avoids environmental pollution to a certain extent.

[0019] In summary, the hemp seed protein peptide obtained by the technical solution provided by the present invention through the combined action of multiple technologies can significantly inhibit the activity of α-amylase and α-glucosidase, thereby blocking the human body's digestion and absorption of starch and sucrose. Clinical test results show that the hemp seed protein peptide prepared by the present invention can effectively reduce the postprandial blood sugar peak and peak area, and the effective sugar-free rate can reach 30~60%. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 This is a graph showing the inhibition of amylase activity by hemp seed protein peptide in Example 3 of the present invention;

[0021] Figure 2 This is a graph showing the inhibition of glucosidase activity by hemp seed protein peptide in Example 3 of the present invention;

[0022] Figure 3 This is a schematic diagram of postprandial blood glucose fluctuation parameters of volunteers in the verification test of the present invention;

[0023] Figure 4 This is a schematic diagram of blood glucose fluctuation parameters after breakfast in continuous blood glucose monitoring of volunteers in the verification test of the present invention;

[0024] Figure 5 This is a schematic diagram of blood glucose fluctuation parameters after afternoon tea during continuous blood glucose monitoring of volunteers in the verification test of the present invention.

[0025] Figure 1 、 2 , 4, and 5 are the abbreviations of hemp seed protein peptide, representing hemp seed protein peptide. DETAILED DESCRIPTION

[0026] To make the objectives, technical solutions, and advantages of the embodiments of the present invention more clear, the technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the described embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field are within the scope of protection of the present invention.

[0027] Example 1

[0028] 1000g of hemp seeds were mechanically shelled and then pulverized. Low-temperature continuous phase change degreasing was performed using No. 15 solvent oil at a solid-liquid ratio of 1:8. The extraction temperature was 20°C, the extraction pressure was 2.5 MPa, and the extraction was carried out at a flow rate of 50 L / h through the extraction kettle. The extraction was continued for 90 minutes. The decomposition temperature was 70°C and the decomposition pressure was 0.5 MPa to obtain defatted hemp meal (oil content ≤3%).

[0029] Defatted hemp meal was mixed with water at a material-to-liquid ratio of 1:10. A complex polysaccharidase (cellulase, pectinase, amylase, and glycosidase, in a ratio of 2:2:1:1) was added at 10‰ of the substrate weight. The pH was adjusted to 3.0 and hydrolysis was performed for 1 hour. The enzymes were inactivated at 85°C for 5 minutes. 2.4 L of Bacillus licheniformis high-efficiency protease was added to the hydrolyzate at 2‰ of the substrate weight, the pH was adjusted to 6.0, and hydrolysis was continued for 15 minutes. The enzymes were inactivated at 85°C for 5 minutes. The extract was centrifuged and filtered, and the solution was cooled to 4°C. Saturated ammonium sulfate solution was continuously added to the resulting extract until the protein flocculated. Salting out was performed for 30 minutes, and the extract was centrifuged and filtered to obtain hemp seed protein peptide. The hemp seed protein peptide was dried and powdered using a spray dryer with an inlet air temperature of 15°C, an outlet air temperature of 30°C, a feed rate of 50 kg / h, and an atomizer pressure of 15 MPa. The hemp seed protein peptide dry powder is obtained and vacuum-sealed in a sterile aluminum foil bag.

[0030] The finished product of hemp seed protein peptide is sterilized in an ultra-high pressure autoclave with a working pressure of 100~500MPa, a working temperature of 25~40℃, a pressure holding time of 20~90s, and is stored at room temperature after sterilization.

[0031] Example 2

[0032] Compared with Example 1, the only difference is that during the polysaccharide enzymatic hydrolysis, the material-liquid ratio is 1:15, and a composite polysaccharide enzyme (cellulase, pectinase, amylase, glycosidase, ratio 2:2:1:1) of 15‰ of substrate mass is added, the pH is adjusted to 5.5, and the hydrolysis treatment is carried out for 4 hours. The enzyme inactivation temperature is 85°C, and the time is 5 minutes. 2.4L of Bacillus licheniformis high-efficiency protease of 2‰ of substrate mass is continued to be added to the hydrolyzate, the pH is adjusted to 6.0, and the hydrolysis treatment is carried out for 15 minutes. The enzyme inactivation temperature is 85°C, and the time is 5 minutes. Centrifugal filtration is performed, the solution is cooled to 4°C, and a saturated ammonium sulfate solution is continuously added to the extract until the protein flocculates, salting out for 90 minutes, and centrifugal filtration is performed to obtain hemp seed protein peptide. The remaining steps and operations are consistent with Example 1.

[0033] Example 3

[0034] Compared with Example 1, the only difference is that during the polysaccharide enzymatic hydrolysis, the material-liquid ratio is 1:20, and a composite polysaccharide enzyme (cellulase, pectinase, amylase, glycosidase, ratio 2:2:1:1) of 10‰ of the substrate mass is added, the pH is adjusted to 4.5, and the hydrolysis treatment is carried out for 3 hours. The enzyme inactivation temperature is 85°C, and the time is 5 minutes. Continue to add 2‰ of the substrate mass of Bacillus licheniformis protease to the hydrolyzate, adjust the pH to 7.5, and carry out hydrolysis treatment for 15 minutes. The enzyme inactivation temperature is 85°C, and the time is 5 minutes. Centrifuge and filter, cool the solution to 4°C, continue to add saturated ammonium sulfate solution to the extract until the protein flocculates, salt out for 30 minutes, and obtain hemp seed protein peptide after centrifugation. The remaining steps and operations are consistent with Example 1.

[0035] Comparative Example 1

[0036] 1000g of hemp seeds were mechanically shelled and then crushed. The hemp meal was defatted using a mechanical oil press and the oil was extracted twice continuously. The defatted hemp meal was mixed with water at a solid-liquid ratio of 1:20. A complex polysaccharidase (cellulase, pectinase, amylase, and glycosidase, in a ratio of 2:2:1:1) was added at 10‰ of the substrate weight. The pH was adjusted to 4.5 and hydrolysis was carried out for 3 hours. The enzyme was inactivated at 85°C for 5 minutes. 2.4L of Bacillus licheniformis high-efficiency protease was added to the hydrolyzate at 2‰ of the substrate weight. The pH was adjusted to 7.5 and hydrolysis was carried out for 15 minutes. The enzyme was inactivated at 85°C for 5 minutes. The solution was centrifuged and filtered, and the solution was cooled to 4°C. Saturated ammonium sulfate solution was continuously added to the resulting extract until the protein flocculated. Salting out occurred for 30 minutes, and the extract was centrifuged and filtered to obtain hemp seed protein peptide. The hemp seed protein peptide liquid was dried and powdered using a spray dryer with an inlet air temperature of 15°C, an outlet air temperature of 30°C, a feed rate of 50 kg / h, and an atomizer pressure of 15 MPa. The obtained hemp seed protein peptide dry powder was vacuum-sealed in sterile aluminum foil bags.

[0037] The finished product of hemp seed protein peptide is sterilized in an ultra-high pressure autoclave with a working pressure of 100~500MPa, a working temperature of 25~40℃, a pressure holding time of 20~90s, and is stored at room temperature after sterilization.

[0038] Comparative Example 2

[0039] 1000g of hemp seeds were mechanically shelled and then pulverized. Low-temperature continuous phase change degreasing was performed using No. 15 solvent oil at a solid-liquid ratio of 1:10. The extraction temperature was 30°C, the extraction pressure was 3.2 MPa, and the extraction was carried out at a flow rate of 50 L / h through the extraction kettle. The extraction was continued for 90 minutes. The decomposition temperature was 70°C and the decomposition pressure was 0.7 MPa to obtain defatted hemp meal (oil content ≤3%).

[0040] Defatted hemp meal was mixed with water at a solid-liquid ratio of 1:20. A complex polysaccharidase (cellulase, pectinase, amylase, and glycosidase, in a ratio of 2:2:1:1) was added at 10‰ of the substrate weight. The pH was adjusted to 4.5, and hydrolysis was performed for 3 hours. The enzymes were inactivated at 85°C for 5 minutes. The extract was then centrifuged and filtered, and saturated ammonium sulfate solution was continuously added to the resulting extract until the protein flocculated. Salting out occurred for 30 minutes, and the extract was centrifuged and filtered to obtain hemp seed protein peptide. The hemp seed protein peptide solution was then dried and powdered using a spray dryer with an inlet air temperature of 15°C, an outlet air temperature of 30°C, a feed rate of 50 kg / h, and an atomizer pressure of 15 MPa. The resulting hemp seed protein peptide powder was vacuum-sealed in sterile aluminum foil bags.

[0041] The finished product of hemp seed protein peptide is sterilized in an ultra-high pressure autoclave with a working pressure of 100~500MPa, a working temperature of 25~40℃, a pressure holding time of 20~90s, and is stored at room temperature after sterilization.

[0042] Examples 1 to 3 were tested to obtain the hemp seed protein peptide yield, α-amylase inhibitory activity, and α-glucosidase inhibitory activity. The data are shown in Table 1.

[0043] Table 1 Data on the yield of hemp seed protein peptides, α-amylase inhibitory activity, and α-glucosidase inhibitory activity of Examples 1 to 3

[0044]

[0045] As can be seen from Table 1, the hemp seed protein peptides prepared in Examples 1 to 3 all have the effect of blocking sugar absorption, and the hemp seed protein peptides prepared by the specific parameters of each process described in Example 3 have the highest yield and the best inhibition of α-amylase and glucosidase activity. It is preliminarily proved that this method is the optimal process for preparing hemp seed protein peptides with sugar absorption blocking activity.

[0046] The hemp seed protein peptides prepared in Example 3 and Comparative Examples 1-2 were tested for hemp seed protein peptide yield, oil content, and sample storage stability. The data are shown in Table 2.

[0047] Table 2 Example 3 and Comparative Examples 1-2 Hemp Seed Protein Peptide Yield, Oil Content, Sample Storage Stability Test Data

[0048]

[0049] As shown in Table 2, the low-temperature continuous phase change defatting and the step-by-step hydrolysis of polysaccharidase and protease used in the present invention have a certain influence on the yield of hemp seed protein peptides, oil content and sugar content.

[0050] Mass spectrometry analysis, in vitro enzyme inhibition experiments and clinical experiments were performed on the hemp seed protein peptide obtained in Example 3 to verify its sugar absorption blocking activity:

[0051] 1. Mass spectrometry analysis of hemp seed protein peptide dry powder was performed using HPLC-MS. The specific parameters and steps were as follows: a reversed-phase column was used, the aqueous phase was 0.1% formic acid aqueous solution, the organic phase was acetonitrile mobile phase, the flow rate was 0.2-1.0 ml / min, the column temperature was 25-40°C, the ESI spray voltage was 2.5-5 kV, the gas was nitrogen, the pressure was 1-3 Pa, and the collision energy was 10-50 eV. The core peptide segment is shown below:

[0052] SEQIDNO: 1: Met-Thr-Met-Trp, SEQIDNO: 2: Tyr-Lys-Pro-Val-Tyr, SEQIDNO: 3: Thr-Pro-Val-Ser-Ile-Leu, SEQIDNO: 4: Met-Ala-His-Leu-Phe, SEQIDNO: 5: Met-Val-Ser-Leu-Tyr.

[0053] 1. Inhibitory activity of α-amylase and glucosidase

[0054] 1.1 α-amylase inhibitory activity assay

[0055] Figure 1 The figure is the assay chart of α-amylase inhibition activity. The IC50 of hemp seed protein peptide was 4.975 mg / mL. Figure 1 It can be observed that the inhibition rate of hemp seed protein peptide on α-amylase activity reached the highest level (84.2%) at a concentration of 4 mg / mL. The results showed that hemp seed protein peptide has a certain inhibitory activity on amylase, which can hinder the hydrolysis and digestion of carbohydrates in food, thereby reducing the body's intake of sugar.

[0056] 1.2 Glucosidase inhibitory activity assay

[0057] Figure 2 The figure is a graph showing the α-glucosidase inhibitory activity. The IC50 of hemp seed protein peptide was 0.53 mg / mL, while the IC50 of hemp seed protein was too large to be calculated by the software. Figure 2 It can be observed that the inhibition rate of hemp protein peptide on α-glucosidase activity reached the highest level (75.1%) at a concentration of 4 mg / mL, which is much higher than the inhibition rate of hemp protein on α-glucosidase, indicating that hemp peptide at an appropriate concentration can effectively delay the absorption of carbohydrates and reduce postprandial hyperglycemia.

[0058] 2. Clinical trials

[0059] Volunteer Recruitment

[0060] Inclusion criteria were: healthy male or female volunteers; age, 18 to 55 years; body mass index, 18 to 25 kg / m2 (healthy group) or BMI>25 (overweight group); no previous diagnosis of diabetes or prediabetes; fasting blood glucose, 3.9 to 5.5 mmol / L (70-100 mg / dL) (healthy group) or fasting blood glucose greater than 6.0 (hyperglycemia group); not taking any medication for diabetes or insulin resistance; and willingness to provide informed written consent to participate in the study.

[0061] Exclusion criteria were: abnormal results of liver function tests (alanine aminotransferase and aspartate aminotransferase), renal function tests (blood urea nitrogen and creatinine), complete blood count, urinalysis, fecal occult blood test, or electrocardiogram; pregnant or breastfeeding women; recent participation in a clinical trial (within the past 3 months); history of malignancy or current malignancy; history of drug dependence, substance abuse, or alcoholism; history of drug or food allergy or allergy to any component of the test product; known history of primary disease of a major organ (such as the heart, liver, or kidney), or a history of digestive, metabolic, neurological, or psychiatric disease; use of medications known to cause organ damage within the previous 3 months; and use of any medications within the past 2 weeks.

[0062] 2.2 Test operation procedures

[0063] All volunteers avoided drinking alcohol or participating in intense physical activities the day before. Before the test, Sinocare continuous blood glucose meters were installed to ensure that all volunteers' instruments were activated, the data was stable, and a family group was established. The formal experiment began on the second day. Fasting for at least 10 hours before the test (drinking water was allowed), and the test began around 8:00 am on the second to fifth days. Volunteers ate a bowl of spinach and egg porridge, a slice of white bread, and a small amount of pickled mustard within 10 minutes. At the same time, hemp protein peptide 2g, 5g, 10g and 1g were taken daily in sequence.

[0064] The second round of testing began after blood sugar returned to baseline after lunch every afternoon. Each person drank a large cup of Starbucks Golden Roast Peach Oatmeal Latte every day, and also consumed 2g, 5g, 10g and 1g of hemp seed protein peptides every day.

[0065] After each test, participants fasted for 2 hours after eating and refrained from food or water. Blood sugar levels after meals were collected every 3 minutes via the "AiKan Health" app's family group.

[0066] 2.3 Area under the Postprandial Glucose Response Curve and Statistical Analysis

[0067] The blood glucose curve was drawn 120 minutes after the meal, and the area under the curve (AUC) was calculated using Graphpad (9.0) software. Repeated measures one-way analysis of variance and paired T test were used to compare the AUC, blood glucose peak, peak time, and maximum blood glucose increase of each intervention with the baseline. Figure 3 (Figure 2 is a schematic diagram of postprandial blood glucose fluctuation parameters for continuous blood glucose monitoring). Paired sample T-tests were used to analyze the differences between different doses and baseline.

[0068] Figure 4 To continuously monitor blood glucose fluctuations after breakfast (8 people in each group, 2 groups in total). Figure 4 It can be seen that the test subjects in the experimental group who consumed hemp protein peptides at the same time as porridge and bread had less blood sugar fluctuations than the control group who only consumed porridge and bread. The statistical analysis of all subjects showed that: taking 1g of hemp protein peptides at the same time as porridge and bread had a certain effect on blood sugar peak and peak area (an average reduction of 30%); after taking 2g of hemp protein peptides, the blood sugar peak of 5 test subjects was significantly reduced (an average reduction of 38%), and the blood sugar peak area was significantly reduced (an average reduction of 49%). After taking 5g of hemp protein peptides, the blood sugar peak of 6 out of 8 test subjects was significantly reduced (an average reduction of 67%), and the blood sugar peak area of these 5 test subjects was significantly reduced (an average reduction of 56%).

[0069] Depend on Figure 5 Continuous blood glucose monitoring was conducted to measure blood glucose fluctuations after afternoon tea (8 participants per group, 2 groups in total). It can be seen that the experimental group, who consumed hemp seed protein peptides while consuming milk tea, had less blood glucose fluctuations than the control group, who consumed milk tea alone. Statistical analysis of all subjects showed that after consuming milk tea, consuming 2g and 5g of hemp seed protein peptides reduced blood glucose fluctuations within 150 minutes after the meal, with lower blood glucose peaks (average decreases of 41.8% and 44.9%, respectively) and a smaller area under the blood glucose curve (average decreases of 61% and 58%, respectively).

[0070] In summary, the results of in vitro enzyme inhibition activity tests and human clinical trials show that the hemp seed protein peptide obtained by the technical solution and preparation method of the present invention can significantly inhibit the activity of α-amylase and α-glucosidase, thereby blocking the human body's digestion and absorption of starch and sucrose. The clinical test results show that the hemp seed protein peptide prepared by the present invention can effectively reduce the postprandial blood sugar peak and peak area, and the effective sugar-free rate can reach 30~60%.

[0071] The above is a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A method for preparing hemp seed protein peptide having the effect of blocking sugar absorption, characterized in that: The following steps are involved: The hemp seeds are crushed and defatted through low-temperature continuous phase change to obtain defatted hemp meal. The defatted hemp meal is hydrolyzed by polysaccharidase and protease in sequence. After the hydrolysis, it is subjected to low-temperature salting out, dried and sterilized to obtain hemp seed protein peptide. The hemp seed protein peptide includes the following short peptide sequences: SEQ ID NO: 1: Met-Thr-Met-Trp, SEQ ID NO: 2: Tyr-Lys-Pro-Val-Tyr, SEQ ID NO: 3: Thr-Pro-Val-Ser-Ile-Leu, SEQ ID NO: 4: Met-Ala-His-Leu-Phe, SEQ ID NO: 5: Met-Val-Ser-Leu-Tyr.

2. The method for preparing the hemp seed protein peptide having the sugar absorption blocking effect according to claim 1, wherein The extraction agent of the low-temperature continuous phase change is No. 15 solvent oil; the mass volume ratio of the hemp seed to the extraction agent is 1:8-15; the low-temperature continuous phase change includes the following conditions: temperature of 20°C-35°C, pressure of 2.5Mpa-4Mpa, flow rate of 50-80L / h, continuous extraction time of 90min-120min, decomposition temperature of 70°C-85°C, and decomposition pressure of 0.5Mpa-1Mpa.

3. The method for preparing the hemp seed protein peptide having the sugar absorption blocking effect according to claim 1, wherein The polysaccharide enzymatic hydrolysis comprises the following steps: defatted hemp meal and water are mixed at a material-liquid ratio of 1:10-30, the mass of the added polysaccharide enzyme is 10-15‰ of the mass of the substrate, the pH is adjusted to 3.0-5.5, and the hydrolysis time is 1-4 hours; the substrates are defatted hemp meal and water.

4. The method for preparing the hemp seed protein peptide having the sugar absorption blocking effect according to claim 1, wherein The polysaccharidase is one or a combination of two or more of cellulase, pectinase, amylase, glycosidase, hemicellulose, glucanase and xylanase.

5. The method for preparing the hemp seed protein peptide having the effect of blocking sugar absorption according to claim 1, wherein Protease hydrolysis involves the following steps: The product after enzymatic hydrolysis of the polysaccharide is inactivated, and then protease is added in an amount of 2-6‰ of the mass of the product after enzymatic hydrolysis of the polysaccharide. The pH is adjusted to 6.0-8.5 for hydrolysis, and the hydrolysis time is 15-35 minutes. The protease is Bacillus licheniformis protease.

6. The method for preparing the hemp seed protein peptide having the effect of blocking sugar absorption according to claim 1, wherein The low-temperature salting out is to centrifuge and filter the product after protease hydrolysis to obtain a filtrate, cool it to 2-4°C, continuously add saturated ammonium sulfate solution to the filtrate until the protein flocculates, salt out for 30-90 minutes, and collect the precipitate after centrifugation.

7. The method for preparing the hemp seed protein peptide having the effect of blocking sugar absorption according to claim 1, wherein The drying is spray drying, which includes the following steps: using a spray dryer for spray drying, with an air inlet temperature of 15-25°C, an air outlet temperature of 30-40°C, a feed rate of 50-150 kg / h, an electrostatic generator voltage of 30-55 kV, and an atomizer pressure of 15-20 MPa.

8. The method for preparing the hemp seed protein peptide having the effect of blocking sugar absorption according to claim 1, wherein The sterilization is ultra-high pressure sterilization, which includes the following steps: the working pressure during ultra-high pressure sterilization is 100-500 MPa, the working temperature is 25-40° C., and the pressure holding time is 20-90 seconds.

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