Wheat gluten based functional food helpful for maintaining healthy level of blood sugar and preparation method of wheat gluten based functional food

Abstract

The invention discloses a vital gluten-based functional food helpful for maintaining the healthy level of blood sugar and a preparation method thereof, and relates to the technical field of biology. The vital wheat gluten-based functional food comprises the following raw material components in parts by weight: 18-22 parts of wheat vital wheat gluten peptide, 8-12 parts of whey protein powder, 4-6 parts of pumpkin powder, 4-6 parts of Chinese yam powder, 2-4 parts of celery powder and 0.4-0.6 part of barley polypeptide. The barley polypeptide is DM-1 polypeptide and / or DM-2 polypeptide; the amino acid sequence of the DM-1 polypeptide is as shown in SEQ ID NO. 1; the amino acid sequence of the DM-2 polypeptide is as shown in SEQ ID NO. 2. According to the invention, specific polypeptides and natural plant components are innovatively combined, so that the functional food for blood glucose management with scientific basis and practical application value is successfully developed, and the functional food has important practical application value.

Description

Technical Field

[0001] The present invention relates to the field of biotechnology, and particularly to a gluten-based functional food that helps maintain a healthy blood glucose level and a method for preparing the same. Background Art

[0002] With the changes in modern lifestyles, diabetes and its related metabolic syndromes have become increasingly serious health problems. The long-term abnormal blood glucose level not only affects the quality of life of patients but also may cause various complications such as cardiovascular diseases, kidney diseases, and retinopathy. Therefore, it is of great significance to develop functional foods that can effectively maintain a healthy blood glucose level.

[0003] Traditionally, drug treatment is the main means of controlling blood glucose, but its side effects and economic burden limit its widespread application. In recent years, bioactive peptides from natural sources have received extensive attention due to their safety and high efficiency. Research has shown that polypeptides with certain specific amino acid sequences can regulate blood glucose levels by promoting insulin secretion, improving insulin sensitivity, or directly inhibiting α-glucosidase.

[0004] Most of the current functional foods on the market have problems such as single composition and unstable effects. Therefore, the present invention aims to provide a composite functional food based on wheat gluten peptides and other natural ingredients to achieve a more comprehensive and lasting blood glucose management effect. Summary of the Invention

[0005] The object of the present invention is to provide a gluten-based functional food that helps maintain a healthy blood glucose level and a method for preparing the same to solve the problems existing in the above-mentioned prior art. This gluten-based functional food helps maintain a healthy blood glucose level and has important practical application value.

[0006] To achieve the above object, the present invention provides the following solutions:

[0007] The present invention provides a gluten-based functional food that helps maintain a healthy blood glucose level, and the raw materials include the following components in parts by weight:

[0008] 18 - 22 parts of wheat gluten peptides, 8 - 12 parts of whey protein powder, 4 - 6 parts of pumpkin powder, 4 - 6 parts of yam powder, 2 - 4 parts of celery powder, and 0.4 - 0.6 parts of barley polypeptide;

[0009] The barley polypeptide is DM-1 polypeptide and / or DM-2 polypeptide;

[0010] The amino acid sequence of the DM-1 polypeptide is as shown in SEQ ID NO.1; the amino acid sequence of the DM-2 polypeptide is as shown in SEQ ID NO.2.

[0011] Preferably, the raw materials include the following components in parts by weight:

[0012] 20 parts of wheat gluten peptide, 10 parts of whey protein powder, 5 parts of pumpkin powder, 5 parts of yam powder, 3 parts of celery powder, and 0.5 part of barley polypeptide.

[0013] Furthermore, the wheat gluten peptide is obtained by subjecting wheat gluten to an enzymatic hydrolysis reaction.

[0014] Furthermore, the enzymes used in the enzymatic hydrolysis reaction are pepsin and papain.

[0015] The present invention also provides a method for preparing the above-mentioned gluten-based functional food, which includes the step of uniformly mixing the raw materials to prepare the gluten-based functional food.

[0016] The present invention also provides a polypeptide that helps maintain a healthy blood glucose level, and the amino acid sequence of the polypeptide is as shown in SEQ ID NO.1 or SEQ ID NO.2.

[0017] The present invention also provides a polypeptide combination that helps maintain a healthy blood glucose level, including the polypeptides with amino acid sequences as shown in SEQ ID NO.1 and SEQ ID NO.2.

[0018] The present invention also provides the use of the above-mentioned polypeptide or polypeptide combination in the preparation of a functional food for maintaining a healthy blood glucose level.

[0019] The present invention also provides the use of the above-mentioned polypeptide or polypeptide combination in the preparation of an α-glucosidase inhibitor.

[0020] The present invention also provides an α-glucosidase inhibitor, and the active ingredient includes the above-mentioned polypeptide or polypeptide combination.

[0021] The present invention discloses the following technical effects:

[0022] The present invention identified two barley polypeptides with α-glucosidase inhibitory activity by liquid chromatography-mass spectrometry (LC-MS), named DM-1 and DM-2. Through experiments, it was confirmed that polypeptides DM-1 and DM-2 have good α-glucosidase inhibitory activity, and can delay the decomposition of carbohydrates and reduce the postprandial blood glucose peak by inhibiting the activity of α-glucosidase, thereby playing a significant blood glucose regulation role.

[0023] The present invention further enhances the overall effect of maintaining a healthy blood glucose level by scientifically compounding a variety of natural ingredients. Among them, wheat gluten peptide, as an active ingredient, provides rich bioactive peptides, which helps to maintain blood glucose balance; whey protein powder is rich in branched-chain amino acids (BCAAs), which can stimulate insulin secretion and participate in muscle energy metabolism; pumpkin powder and yam powder are rich in dietary fiber and plant polysaccharides, which can delay gastric emptying and stabilize blood glucose fluctuations; celery powder contains flavonoids, which have antioxidant and anti-inflammatory effects and indirectly support blood glucose management; barley polypeptides (DM-1 and DM-2) form a synergistic effect with wheat gluten peptide to enhance the blood glucose lowering function.

[0024] The present invention successfully develops a functional food for blood glucose management with both scientific basis and practical application value by innovatively combining specific polypeptides with natural plant ingredients, which has important practical application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required to be used in the embodiments. Obviously, the drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0026] Figure 1 It is a detection result graph of the blood glucose level of mice after intragastric administration of barley polypeptide;

[0027] Figure 2 It is a detection result graph of the blood glucose level of mice in the meal replacement experiment of Example 10. DETAILED DESCRIPTION OF THE INVENTION

[0028] Now, various exemplary embodiments of the present invention will be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, characteristics, and implementation schemes of the present invention.

[0029] It should be understood that the terms described in the present invention are only for describing specific embodiments and are not used to limit the present invention. In addition, for the numerical ranges in the present invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any intermediate value within any stated value or stated range, as well as each smaller range between any other stated value or intermediate value within the stated range, is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded from the range.

[0030] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Although this invention only describes preferred methods and materials, any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of this invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials related to the said documents. In case of conflict with any incorporated document, the content of this specification shall prevail.

[0031] Without departing from the scope or spirit of this invention, various improvements and changes can be made to the specific embodiments of the description of this invention, which will be obvious to those skilled in the art. Other embodiments obtained from the description of this invention will be obvious to those skilled in the art. The description and examples of this invention are merely exemplary.

[0032] Regarding the use of "comprising", "including", "having", "containing", etc. in this article, they are all open-ended terms, meaning including but not limited to.

[0033] The preparation method of wheat gluten peptide used in the following examples is as follows:

[0034] Add 9 times the weight of water to wheat gluten powder, and then add pepsin and papain for enzymatic hydrolysis. Among them, pepsin and papain are added according to 0.2% and 0.3% of the weight of wheat gluten powder respectively. The temperature of the enzymatic hydrolysis reaction is 55°C and the pH is 6.5. After enzymatic hydrolysis for 4 h, the temperature is raised to 95°C to inactivate the enzyme for 15 min. Filter the enzymatic hydrolysate to obtain the clear liquid, and then concentrate and spray-dry it to obtain wheat gluten peptide.

[0035] Example 1

[0036] (1) Mix barley flour and 0.2 wt% NaOH aqueous solution evenly according to a volume ratio of 1:500, continuously stir at 50°C for 1 h, then centrifuge to obtain the supernatant, adjust the pH value of the supernatant to 4.0 with HCl solution, let it stand for 20 min, and then centrifuge at 4°C to obtain the precipitate.

[0037] (2) Take out the precipitate obtained in step (1), add deionized water three times the weight of the precipitate, adjust the pH value of the mixture to 6.0, and then raise the temperature to 50°C. Then, add pepsin according to 0.5 wt% of the weight of the precipitate, and carry out an oscillating enzymatic hydrolysis reaction at 50°C at a rotation speed of 150 r / min for 4 hours. After the enzymatic hydrolysis is completed, raise the temperature to 100°C and keep it for 10 minutes to inactivate the enzyme activity, and then cool naturally. Finally, centrifuge the enzyme-inactivated mixture and take the supernatant to obtain the enzymatic hydrolysate.

[0038] (3) Ultrafiltration separation is carried out on the enzymatic hydrolysate obtained in step (2), and the retentate in the range of 500 Da - 3 kDa is collected.

[0039] (4) The 500 Da - 3 kDa fraction is subjected to reduction alkylation using dithiothreitol and iodoacetamide, and desalted using a self-packed desalting column. The obtained sample is loaded onto an LC column (0.15 mm × 150 mm, RP-C18, Column Technology Inc.). Among them, mobile phase A: 0.1% formic acid aqueous solution, mobile phase B: 0.1% formic acid - 80% acetonitrile aqueous solution, flow rate: 600 nL / min, total analysis time: 66 min. The relevant LC gradient program is as follows: 0 - 2 min, linear gradient of mobile phase B from 4% to 8%; 2 - 35 min, linear gradient of mobile phase B from 8% to 28%; 35 - 55 min, linear gradient of mobile phase B from 28% to 40%; 55 - 56 min, linear gradient of mobile phase B from 40% to 95%; 56 - 66 min, mobile phase B is maintained at 95%. The sample is used to collect data by LC-MS / MS to generate a mass spectrometry Raw file, which is opened with Xcalibur and retrieved by the software PEAKSStudio10.6 to obtain the total ion current chromatogram and polypeptide sequence of the sample.

[0040] (5) Detection of α-glucosidase inhibitory activity is carried out on the identified barley polypeptides DM-1 and DM-2 (amino acid sequences are shown in Table 1).

[0041] Polypeptides DM-1 and DM-2 are synthesized by solid-phase synthesis method. Polypeptides DM-1 and DM-2 are respectively formulated into solutions with a concentration of 0.1 mg / mL using PBS (0.2 M, pH 6.8), and then their inhibitory activities against α-glucosidase are detected respectively. The detection method is as follows:

[0042] The experiment is set up into four groups, namely: control group (A0), sample group (A1), blank group (A2), and sample blank group (A3). In the enzyme-labeled plate, 20 μL of the sample, 50 μL of PBS (0.2 M, pH 6.8), and 50 μL of p-nitrophenyl-α-D-glucopyranoside (pNPG, 1 mg / mL) are added in sequence, reacted at 37 °C for 10 min, then 50 μL of α-glucosidase (1 U / mL) is added, reacted at 37 °C for 30 min, and finally 80 μL of Na2CO3 (0.2 mol / L) is added to terminate the reaction. The absorbance value is measured at 405 nm. The control group is PBS + enzyme + pNPG + Na2CO3; the sample group is sample + enzyme + pNPG + Na2CO3; the blank group is PBS + PBS + pNPG + Na2CO3; the sample blank group is sample + PBS + PBS + Na2CO3. The inhibition rate calculation formula is as follows:

[0043] Inhibition rate (%) of α-glucosidase = [1 - (A1 - A3) / (A0 - A2)] × 100%.

[0044] The results of the detection of α-glucosidase inhibitory activity of barley polypeptides DM-1 and DM-2 are shown in Table 2. The results show that both barley polypeptides DM-1 and DM-2 have α-glucosidase inhibitory activity.

[0045] Table 1 Amino acid sequences of barley polypeptides

[0046] Polypeptide Name Amino Acid Sequence Sequence Number DM-1 reqldsqdk SEQ ID NO.1 DM-2 viravivrtcke SEQ ID NO.2

[0047] Table 2 Inhibition rates of α-glucosidase of barley polypeptides

[0048] Polypeptide Name α-Glucosidase Inhibition Rate DM-1 63.81％ DM-2 55.27％

[0049] Example 2

[0050] 1. Experimental grouping

[0051] Inject alloxan solution into the caudal vein of mice at a dose of 150 mg / kg body weight. Under the action of alloxan, the pancreatic islet β cells of mice are damaged, resulting in insulin secretion dysfunction, thereby constructing an experimental diabetic mouse model. One week after the establishment of the diabetic mouse model, 80 model mice are taken and randomly divided into 4 groups on average, namely the model group, the positive control group and two polypeptide groups. At the same time, 20 normal mice with the same background are selected as the normal control group. Each group of mice is given intragastric administration according to Table 3, once a day for 4 consecutive weeks.

[0052] Table 3 Mouse experimental design table

[0053] Group Agent Single Oral Gavage Dose Normal Control Group Normal Saline 0.3mL Model Group Normal Saline 0.3mL Positive Control Group Metformin Hydrochloride 0.5mL(120mg / kg) DM-1 Group Polypeptide DM-1 0.5mL(200mg / kg) DM-2 Group Polypeptide DM-2 0.5mL(200mg / kg)

[0054] 2. Determination of oral glucose tolerance (OGTT)

[0055] After 4 weeks of intragastric administration, after the last intragastric administration, fast the mice for 8 h without water, measure the fasting blood glucose (FBG), and 30 min later, give each mouse an intragastric administration of 2 g / kg glucose solution, and collect blood samples from the caudal vein to measure the blood glucose value (BG) at 0.5 h, 1 h, 1.5 h and 2 h respectively. The results are shown in Figure 1 .

[0056] According to Figure 1It can be seen that after intragastric administration of glucose solution to each group of mice, the blood glucose values all showed a trend of first rising and then falling. The blood glucose values of the normal control group of mice could return to the normal level within 2 h, while the blood glucose values of the remaining groups were still significantly higher than those of the normal control group after 2 h. Compared with the model group, the blood glucose values of the DM-1 group, DM-2 group and positive control group were all at lower levels within 2 h. The above results indicate that the intake of polypeptides DM-1 and DM-2 can improve the glucose tolerance of diabetic mice, inhibit the increase of postprandial blood glucose and maintain blood glucose stability.

[0057] Example 3

[0058] A gluten-based functional food, with the following raw materials:

[0059] 20 g of wheat gluten peptide, 10 g of whey protein powder, 5 g of pumpkin powder, 5 g of yam powder, 3 g of celery powder and 0.5 g of barley polypeptide DM-1.

[0060] Mix the raw materials evenly to obtain the gluten-based functional food.

[0061] Example 4

[0062] A gluten-based functional food, with the following raw materials:

[0063] 18 g of wheat gluten peptide, 12 g of whey protein powder, 4 g of pumpkin powder, 6 g of yam powder, 2 g of celery powder and 0.6 g of barley polypeptide DM-1.

[0064] Mix the raw materials evenly to obtain the gluten-based functional food.

[0065] Example 5

[0066] A gluten-based functional food, with the following raw materials:

[0067] 22 g of wheat gluten peptide, 8 g of whey protein powder, 6 g of pumpkin powder, 4 g of yam powder, 4 g of celery powder and 0.4 g of barley polypeptide DM-1.

[0068] Mix the raw materials evenly to obtain the gluten-based functional food.

[0069] Example 6

[0070] A gluten-based functional food, with the following raw materials:

[0071] 20 g of wheat gluten peptide, 10 g of whey protein powder, 5 g of pumpkin powder, 5 g of yam powder, 3 g of celery powder and 0.5 g of barley polypeptide DM-2.

[0072] Mix the raw materials evenly to obtain the gluten-based functional food.

[0073] Example 7

[0074] A gluten-based functional food, with the following raw materials:

[0075] 18 g of wheat gluten peptide, 12 g of whey protein powder, 4 g of pumpkin powder, 6 g of Chinese yam powder, 2 g of celery powder, and 20.6 g of barley polypeptide DM-2.

[0076] Mix all the raw materials evenly to obtain the gluten-based functional food.

[0077] Example 8

[0078] A gluten-based functional food, with the following raw materials:

[0079] 22 g of wheat gluten peptide, 8 g of whey protein powder, 6 g of pumpkin powder, 4 g of Chinese yam powder, 4 g of celery powder, and 20.4 g of barley polypeptide DM-2.

[0080] Mix all the raw materials evenly to obtain the gluten-based functional food.

[0081] Example 9

[0082] A gluten-based functional food, with the following raw materials:

[0083] 20 g of wheat gluten peptide, 10 g of whey protein powder, 5 g of pumpkin powder, 5 g of Chinese yam powder, 3 g of celery powder, 11.5 g of barley polypeptide DM-1, and 1.5 g of barley polypeptide DM-2.

[0084] Mix all the raw materials evenly to obtain the gluten-based functional food.

[0085] Comparative Example 1

[0086] A gluten-based functional food, with the following raw materials:

[0087] 23 g of wheat gluten peptide, 10 g of whey protein powder, 5 g of pumpkin powder, 5 g of Chinese yam powder, and 3 g of celery powder.

[0088] Mix all the raw materials evenly to obtain the gluten-based functional food.

[0089] Example 10

[0090] 1. Experimental grouping

[0091] Inject alloxan solution into the caudal vein of mice at a dose of 150 mg / kg body weight. Under the action of alloxan, the pancreatic islet β-cells of mice are damaged, resulting in insulin secretion dysfunction, thereby constructing an experimental diabetic mouse model. One week after the establishment of the diabetic mouse model, 100 model mice were taken and randomly divided into 5 groups on average, namely the model group and 4 meal replacement groups. At the same time, 20 normal mice with the same background were selected as the normal control group. The mice in each group were subjected to a meal replacement experiment according to Table 4 for 4 consecutive weeks.

[0092] Table 4 Grouping of Meal Replacement Experiment

[0093] Group Feed Normal Control Group Normal Feed Model Group Normal Feed Group A Replace 50% of the normal feed with the gluten-based functional food of Example 3 Group B Replace 50% of the normal feed with the gluten-based functional food of Example 6 Group C Replace 50% of the normal feed with the gluten-based functional food of Example 9 Group D Replace 50% of the normal feed with the gluten-based functional food of Comparative Example 1

[0094] 2. Determination of Oral Glucose Tolerance

[0095] After 4 weeks of meal replacement, after the last meal replacement, fast the mice for 8 h without water deprivation, and measure the fasting blood glucose (FBG). After 30 min, each mouse was intragastrically administered a 2 g / kg glucose solution, and blood samples were collected from the caudal vein at 0.5 h, 1 h, 1.5 h, and 2 h respectively to measure the blood glucose values. The results are shown in Figure 2 . According to Figure 2 It can be seen that compared with the model group, the blood glucose values of groups A, B, and C were all at lower levels within 2 h, and the blood glucose value of group C was the lowest.

[0096] Based on the experimental results of this example, it can be seen that feeding the gluten-based functional food prepared by the present invention to mice for meal replacement is beneficial to improving the glucose tolerance of diabetic mice, inhibiting the increase of postprandial blood glucose, and maintaining blood glucose stability.

[0097] The above-described embodiments are only descriptions of the preferred embodiments of the present invention, and do not limit the scope of the present invention. Without departing from the design spirit of the present invention, various deformations and improvements made by those of ordinary skill in the art to the technical solutions of the present invention shall fall within the protection scope determined by the claims of the present invention.

Claims

1. A gluten-based functional food that helps maintain a healthy blood sugar level, characterized in that, The raw materials include the following components in parts by weight: 18 - 22 parts of wheat gluten peptide, 8 - 12 parts of whey protein powder, 4 - 6 parts of pumpkin powder, 4 - 6 parts of yam powder, 2 - 4 parts of celery powder, and 0.4 - 0.6 part of barley polypeptide; The barley polypeptide is DM - 1 polypeptide and / or DM - 2 polypeptide; The amino acid sequence of the DM - 1 polypeptide is as shown in SEQ ID NO.1; the amino acid sequence of the DM - 2 polypeptide is as shown in SEQ ID NO.

2.

2. The wheat gluten-based functional food according to claim 1, wherein The raw materials include the following components in parts by weight: 20 parts of wheat gluten peptide, 10 parts of whey protein powder, 5 parts of pumpkin powder, 5 parts of yam powder, 3 parts of celery powder, and 0.5 part of barley polypeptide.

3. The wheat gluten-based functional food according to claim 1 or 2, characterized in that, The wheat gluten peptide is obtained by subjecting wheat gluten to an enzymatic hydrolysis reaction.

4. The gluten-based functional food according to claim 3, wherein The enzymes used in the enzymatic hydrolysis reaction are pepsin and papain.

5. A method for preparing a gluten-based functional food according to any one of claims 1-4, characterized in that, It includes the step of uniformly mixing the raw materials to prepare the gluten - based functional food.

6. A polypeptide that helps maintain a healthy blood glucose level, characterized in that, The amino acid sequence of the polypeptide is as shown in SEQ ID NO.1 or SEQ ID NO.

2.

7. A polypeptide combination that helps maintain a healthy blood glucose level, characterized in that, It includes polypeptides with amino acid sequences as shown in SEQ ID NO.1 and SEQ ID NO.

2.

8. Use of a polypeptide as claimed in claim 6 or a polypeptide combination as claimed in claim 7 in the preparation of a functional food for maintaining a healthy blood glucose level.

9. Use of a polypeptide as claimed in claim 6 or a polypeptide combination as claimed in claim 7 in the preparation of an α - glucosidase inhibitor.

10. An α-glucosidase inhibitor, characterized in that, The active ingredient includes the polypeptide as claimed in claim 6 or the polypeptide combination as claimed in claim 7.

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