A novel spirulina-derived pentapeptide LPELY

Abstract

The application discloses a novel spirulina-derived pentapeptide LPELY, belongs to the technical field of biotechnology, and relates to a spirulina-derived pentapeptide, which is purified from spirulina and has an amino acid sequence as shown in the following: Leu-Pro-Glu-Leu-Tyr, abbreviated as LPELY, and a molecular weight of 633.72 Da. The novel spirulina-derived pentapeptide LPELY obtained in the application has a significant inhibitory effect on pancreatic lipase and also has strong free radical scavenging activity. When the novel spirulina-derived pentapeptide LPELY is applied to the preparation of a blood lipid-lowering drug, the drug can have good blood lipid-lowering effect. The novel spirulina-derived pentapeptide LPELY can also be applied to cosmetics as an antioxidant active ingredient, so that the cosmetics have excellent antioxidant function. The novel spirulina-derived pentapeptide LPELY can also be applied to food preparation, so that the food has excellent blood lipid-lowering and antioxidant effects.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, and more specifically, to a novel spirulina-derived pentapeptide LPELY, its purification method, and its applications. Background Technology

[0002] With the improvement of people's living standards and changes in dietary habits, the incidence of hyperlipidemia and its complications is also increasing. Pancreatic lipase is the main lipase secreted by the pancreas. It can hydrolyze dietary lipids in the digestive system, converting triglyceride substrates in ingested fats into monoglycerides and free fatty acids. These hydrolysis products are absorbed by the epithelial cells of the small intestine, thus directly affecting the absorption of fatty acids in the intestine. Pancreatic lipase inhibitors can bind to the active lipases in the pancreas and small intestine, altering the conformation of trypsin and inhibiting its catalytic activity, thereby inhibiting the hydrolysis of lipids in food, reducing lipid digestion and absorption, and the accumulation of adipose tissue. Researching and developing pancreatic lipase inhibitors or products is crucial for hyperlipidemia and the control of obesity.

[0003] Spirulina is a type of cyanobacteria with physiological functions such as liver protection, antioxidation, antiviral activity, anticancer properties, and immune enhancement. Spirulina protein accounts for 50%–70% of its dry weight, containing 18 amino acids, including all eight essential amino acids for the human body, making it a high-quality protein source. Currently, spirulina is mainly used in powder and tablet form. However, due to its limited solubility, it presents challenges in direct absorption by the human body.

[0004] Bioactive peptides are amino acid polymers that are beneficial or bioactive to the life activities of organisms, also known as functional peptides. Their structure is loose, ranging from two amino acids to hundreds linked by peptide bonds. Due to their wide availability and high safety profile, they have become a research hotspot in recent years. Small molecule peptides are also a class of highly active peptides. They can be absorbed by the body in their intact form; they are actively absorbed with low or no energy consumption; they can directly enter the bloodstream and be delivered to various parts of the body, making them widely used in the biomedical field. Exploring the inhibitory effect of small molecule peptides on pancreatic lipase and determining their lipid-lowering bioactivity is one of the research directions for those skilled in the art. Therefore, the development of small molecule peptides with lipid-lowering activity has attracted much attention. Modern nutrition science shows that proteins are not necessarily absorbed as completely free amino acids after being acted upon by digestive enzymes, but are mainly absorbed in the form of low-peptides, which have higher nutritional value and bioavailability. Currently, enzymatic hydrolysis has become a major method for the industrial production of bioactive peptides, but it suffers from low antioxidant activity, limiting its application. Summary of the Invention

[0005] Therefore, in order to address the problem that existing bioactive peptides have low antioxidant activity and limited applications, this invention provides a novel spirulina-derived pentapeptide, LPELY, along with its purification method and applications. The specific technical solution is as follows:

[0006] A novel spirulina-derived pentapeptide, LPELY, has a molecular weight of 633.72 Da and a sequence of Leu-Pro-Glu-Leu-Tyr. Its chemical structure is as follows:

[0007]

[0008] This invention provides a novel purification method for the spirulina-derived pentapeptide LPELY, the purification method comprising the following steps:

[0009] The novel spirulina-derived pentapeptide LPELY to be purified was loaded onto a chromatographic column; gradient elution was performed using mobile phase A and mobile phase B as eluents, where mobile phase A was an aqueous solution containing 0.1% trifluoroacetic acid by volume, and mobile phase B was an acetonitrile solution containing 0.1% trifluoroacetic acid by volume; the gradient elution program was as follows: the initial volume ratio of mobile phase A was 10%, and from 0.01 min to 25 min after loading, the volume ratio of mobile phase A increased to 10%–70%, and from 25 min to 25.01 min, the volume ratio of mobile phase A increased to 100%, and the process was maintained at 100% until the operation was stopped, with a detection wavelength of 220 nm; the novel spirulina-derived pentapeptide LPELY solution with the target peak was collected.

[0010] Furthermore, the chromatographic column is C18. 18 The chromatographic column; and / or, mobile phase A is an aqueous solution containing 0.1% trifluoroacetic acid by volume, and mobile phase B is an acetonitrile solution containing 0.1% trifluoroacetic acid by volume; and / or, during gradient elution, the initial volume proportion of mobile phase A is 10%, and the volume proportion of mobile phase A increases to 70% within 0.01 min to 25 min after loading, and increases to 100% within 25 min to 25.01 min.

[0011] The present invention also provides an application of a novel spirulina-derived pentapeptide LPELY, wherein the application is the use of the novel spirulina-derived pentapeptide LPELY in the preparation of lipid-lowering drug products.

[0012] The present invention also provides an application of a novel spirulina-derived pentapeptide LPELY, wherein the application is the use of the novel spirulina-derived pentapeptide LPELY in the preparation of cosmetics.

[0013] The present invention also provides an application of a novel spirulina-derived pentapeptide LPELY, wherein the application is the use of the novel spirulina-derived pentapeptide LPELY as a food additive in food preparation.

[0014] The present invention also provides an application of a novel spirulina-derived pentapeptide LPELY, wherein the novel spirulina-derived pentapeptide LPELY is used as a lipid-lowering active ingredient and / or an antioxidant active ingredient in the preparation of cosmetics or food.

[0015] In addition, a composition is provided that contains the novel spirulina-derived pentapeptide LPELY.

[0016] Furthermore, the composition is one of a lipid-lowering drug composition, a cosmetic composition, and a food composition.

[0017] The above-mentioned scheme purifies pentapeptides from spirulina, which have a significant inhibitory effect on pancreatic lipase and strong free radical scavenging activity. When applied to the preparation of lipid-lowering drugs, it can endow the drugs with good lipid-lowering effects. It can also be used as an antioxidant active ingredient in cosmetics, giving cosmetics excellent antioxidant functions. It can also be applied in food preparation, giving food excellent lipid-lowering and antioxidant effects. Attached Figure Description

[0018] Figure 1 HPLC chromatogram for purity identification of LPELY, a novel spirulina-derived pentapeptide;

[0019] Figure 2 This is the ESI-MS spectrum of LPELY, a novel spirulina-derived pentapeptide. The horizontal axis represents m / z (mass-to-charge ratio), and the vertical axis represents intensity (signal strength).

[0020] Figure 3 This is a graph showing the lipid-lowering activity of LPELY, a novel spirulina-derived pentapeptide. The horizontal axis represents the concentration of the tested sample, and the vertical axis represents the pancreatic lipase inhibition rate.

[0021] Figure 4 This is a graph showing the antioxidant activity of LPELY, a novel spirulina-derived pentapeptide. The horizontal axis represents the concentration of the tested sample, and the vertical axis represents the ABTS free radical inhibition rate. Detailed Implementation

[0022] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to its embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and do not limit the scope of protection of the invention.

[0023] Unless otherwise defined, 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. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0024] One embodiment of the present invention discloses a novel spirulina-derived pentapeptide, LPELY, which has a molecular weight of 633.72 Da and a sequence of Leu-Pro-Glu-Leu-Tyr. Its chemical structure is as follows:

[0025]

[0026] In one embodiment, the synthesis of the novel spirulina pentapeptide LPELY includes the following steps: using dichlorotriphenylmethyl resin, following the characteristic of the amino acid sequence Leu-Pro-Glu-Leu-Tyr, the carboxyl group of Tyr is first covalently linked to a resin, then the amino group of Leu and the carboxyl group of Glu undergo a dehydration reaction, after which Pro is added, and the amino group of Leu reacts with the carboxyl group of Pro, and amino acids are added sequentially from right to left. After adding the last Leu amino acid, the resin is then removed to obtain the novel spirulina pentapeptide LPELY.

[0027] In one embodiment, the present invention provides a method for purifying a novel spirulina-derived pentapeptide LPELY, the purification method comprising the following steps:

[0028] The novel spirulina-derived pentapeptide LPELY to be purified was loaded onto a chromatographic column; gradient elution was performed using mobile phase A and mobile phase B as eluents, where mobile phase A was an aqueous solution containing 0.1% trifluoroacetic acid by volume, and mobile phase B was an acetonitrile solution containing 0.1% trifluoroacetic acid by volume; the gradient elution program was as follows: the initial volume ratio of mobile phase A was 10%, and from 0.01 min to 25 min after loading, the volume ratio of mobile phase A increased to 10%–70%, and from 25 min to 25.01 min, the volume ratio of mobile phase A increased to 100%, and the process was maintained at 100% until the operation was stopped, with a detection wavelength of 220 nm; the novel spirulina-derived pentapeptide LPELY solution with the target peak was collected.

[0029] In one embodiment, the chromatographic column is C10. 18The chromatographic column; and / or, mobile phase A is an aqueous solution containing 0.1% trifluoroacetic acid by volume, and mobile phase B is an acetonitrile solution containing 0.1% trifluoroacetic acid by volume; and / or, during gradient elution, the initial volume proportion of mobile phase A is 10%, and the volume proportion of mobile phase A increases to 70% within 0.01 min to 25 min after loading, and increases to 100% within 25 min to 25.01 min.

[0030] In one embodiment, the present invention also provides an application of a novel spirulina-derived pentapeptide LPELY, wherein the application is the use of the novel spirulina-derived pentapeptide LPELY in the preparation of lipid-lowering drug products.

[0031] In one embodiment, the present invention also provides an application of a novel spirulina-derived pentapeptide LPELY, wherein the application is the use of the novel spirulina-derived pentapeptide LPELY in the preparation of cosmetics.

[0032] In one embodiment, the present invention also provides an application of a novel spirulina-derived pentapeptide LPELY, wherein the application is the use of the novel spirulina-derived pentapeptide LPELY as a food additive in food preparation.

[0033] In one embodiment, the present invention also provides an application of a novel spirulina-derived pentapeptide LPELY, wherein the novel spirulina-derived pentapeptide LPELY is used as a lipid-lowering active ingredient and / or an antioxidant active ingredient in the preparation of cosmetics or food.

[0034] In one embodiment, a composition is also provided, the composition containing the novel spirulina-derived pentapeptide LPELY.

[0035] In one embodiment, the composition is one of a lipid-lowering drug composition, a cosmetic composition, and a food composition.

[0036] The above-mentioned method purifies pentapeptides from spirulina, which have a significant inhibitory effect on pancreatic lipase and strong free radical scavenging activity. When applied to the preparation of lipid-lowering drugs, it can endow the drugs with good lipid-lowering effects. It can also be used as an antioxidant active ingredient in cosmetics, giving cosmetics excellent antioxidant functions. It can also be applied in food preparation, giving food excellent lipid-lowering and antioxidant effects.

[0037] The implementation schemes of the present invention will now be described in detail with reference to specific embodiments.

[0038] Example 1:

[0039] Solid-phase synthesis of novel spirulina-derived pentapeptide LPELY: Using dichlorotriphenylmethyl resin (Nanjing Jietai Biotechnology Co., Ltd.), following the characteristic of the amino acid sequence Leu-Pro-Glu-Leu-Tyr, the carboxyl group of Tyr was first covalently linked to a resin. Then, the amino group of Leu and the carboxyl group of Glu underwent a dehydration reaction. After treatment, Pro was added, and the amino group of Leu reacted with the carboxyl group of Pro. Amino acids were added sequentially from right to left. After adding the last Leu amino acid, the resin was removed to obtain the novel spirulina pentapeptide LPELY.

[0040] Purification of a novel spirulina-derived pentapeptide:

[0041] The final product obtained from peptide solid-phase synthesis was purified by high-performance liquid chromatography (HPLC). The purification process used a SHIMADZU Inertsil ODS-SP (4.6 mm × 250 mm, 5 μm) column at a flow rate of 1.0 mL / min. The HPLC system used two channels: solvent A (mobile phase A): an aqueous solution containing 0.1% (v / v) trifluoroacetic acid; solvent B (mobile phase B): an acetonitrile solution containing 0.1% (v / v) trifluoroacetic acid. Gradient elution was performed as follows: the sum of the volume percentages of A and B was 100%, with A initially at 10%. Within 0.01 min to 25 min, the proportion of A increased to 70%, and within 25 min to 25.1 min, the proportion of A increased to 100%. The elution was maintained at 100% for 30 min, and the detection wavelength was 220 nm. The novel spirulina-derived pentapeptide LPELY solution was collected, rapidly cooled with liquid nitrogen, and then lyophilized. A product with a purity of over 98% was obtained. The purity identification HPLC results are shown below. Figure 1 The structure was identified by ESI-MS (e.g.) Figure 2 (As shown). The novel spirulina-derived pentapeptide LPELY was identified as having a molecular weight of 633.72 Da and a sequence of Leu-Pro-Glu-Leu-Tyr. Its chemical structure is as follows:

[0042]

[0043] Example 2:

[0044] A solution of the novel spirulina-derived pentapeptide LPELY from Example 1, at a concentration of 0.0625 mg / mL, was used to determine its inhibitory activity on pancreatic lipase. 50 μL (6 mg / mL) of the novel spirulina-derived pentapeptide LPELY, 25 μL (5 mM) of 4-nitrophenylbutyrate, and 25 μL (1.25 mg / mL) of pancreatic lipase were added to a 96-well plate. The volume was adjusted to 150 μL with PBS, and the plate was incubated at 37°C for 45 min. The absorbance was measured at 405 nm. According to the formula, the inhibition rate of pancreatic lipase was 9.41% when the concentration of the novel spirulina-derived pentapeptide LPELY solution was 0.0625 mg / mL.

[0045] Example 3:

[0046] A 0.125 mg / mL solution of the novel spirulina-derived pentapeptide LPELY from Example 1 was used to determine its pancreatic lipase inhibitory activity. 50 μL (6 mg / mL) of the novel spirulina-derived pentapeptide LPELY, 25 μL (5 mM) of 4-nitrophenylbutyrate, and 25 μL (1.25 mg / mL) of pancreatic lipase were added to a 96-well plate. The volume was adjusted to 150 μL with PBS, and the plate was incubated at 37°C for 45 min. The absorbance was measured at 405 nm. According to the formula, the inhibition rate of pancreatic lipase was 11.34% when the concentration of the novel spirulina-derived pentapeptide LPELY solution was 0.125 mg / mL.

[0047] Example 4:

[0048] A 0.25 mg / mL solution of the novel spirulina-derived pentapeptide LPELY from Example 1 was used to determine its pancreatic lipase inhibitory activity. 50 μL (6 mg / mL) of the novel spirulina-derived pentapeptide LPELY, 25 μL (5 mM) of 4-nitrophenylbutyrate, and 25 μL (1.25 mg / mL) of pancreatic lipase were added to a 96-well plate. The volume was adjusted to 150 μL with PBS, and the plate was incubated at 37°C for 45 min. The absorbance was measured at 405 nm. According to the formula, the inhibition rate of pancreatic lipase was 15.32% when the concentration of the novel spirulina-derived pentapeptide LPELY solution was 0.25 mg / mL.

[0049] Example 5:

[0050] A 0.5 mg / mL solution of the novel spirulina-derived pentapeptide LPELY from Example 1 was used to determine its pancreatic lipase inhibitory activity. 50 μL (6 mg / mL) of the novel spirulina-derived pentapeptide LPELY, 25 μL (5 mM) of 4-nitrophenylbutyrate, and 25 μL (1.25 mg / mL) of pancreatic lipase were added to a 96-well plate. The volume was adjusted to 150 μL with PBS, and the plate was incubated at 37°C for 45 min. The absorbance was measured at 405 nm. According to the formula, the inhibition rate of pancreatic lipase was 23.75% when the concentration of the novel spirulina-derived pentapeptide LPELY solution was 0.5 mg / mL.

[0051] Example 6:

[0052] A 1 mg / mL solution of the novel spirulina-derived pentapeptide LPELY from Example 1 was used to determine its pancreatic lipase inhibitory activity. 50 μL (6 mg / mL) of the novel spirulina-derived pentapeptide LPELY, 25 μL (5 mM) of 4-nitrophenylbutyrate, and 25 μL (1.25 mg / mL) of pancreatic lipase were added to a 96-well plate. The volume was adjusted to 150 μL with PBS, and the plate was incubated at 37°C for 45 min. The absorbance was measured at 405 nm. According to the formula, the inhibition rate of pancreatic lipase was 42.85% when the concentration of the novel spirulina-derived pentapeptide LPELY solution was 1 mg / mL.

[0053] Example 7:

[0054] A 2 mg / mL solution of the novel spirulina-derived pentapeptide LPELY from Example 1 was used to determine its pancreatic lipase inhibitory activity. 50 μL (6 mg / mL) of the novel spirulina-derived pentapeptide LPELY, 25 μL (5 mM) of 4-nitrophenylbutyrate, and 25 μL (1.25 mg / mL) of pancreatic lipase were added to a 96-well plate. The volume was adjusted to 150 μL with PBS, and the plate was incubated at 37°C for 45 min. The absorbance was measured at 405 nm. According to the formula, the inhibition rate of pancreatic lipase was 66.11% when the concentration of the novel spirulina-derived pentapeptide LPELY solution was 2 mg / mL.

[0055] Example 8:

[0056] The antioxidant properties of the novel spirulina-derived pentapeptide LPELY solution and vitamin C solution (12.5 μg / mL, Example 1) were compared and tested separately. The samples were measured as follows: 20 μL of the novel spirulina-derived pentapeptide LPELY solution (12.5 μg / mL) and 20 μL of the vitamin C solution (12.5 μg / mL) were added, along with 180 μL of ABTS working solution. The absorbance value of the sample solution was recorded as Sample A, and the absorbance value of the vitamin C solution was recorded as AVc. A model group (20 μL distilled water and 180 μL ABTS working solution) was set up, with absorbance values ​​recorded as Model A and AVc. A blank group (20 μL distilled water and 180 μL PBS working solution) was set up, with absorbance values ​​recorded as Blank A. According to the formula, when the solution concentration is 12.5 μg / mL, the ABTS radical scavenging rate of the novel spirulina-derived pentapeptide LPELY solution is 91.57%, and the ABTS radical scavenging rate of the vitamin C solution is 100%.

[0057] Example 9:

[0058] The antioxidant properties of the novel spirulina-derived pentapeptide LPELY solution and vitamin C solution (25 μg / mL, Example 1) were compared and tested separately. The samples were measured as follows: 20 μL of the novel spirulina-derived pentapeptide LPELY solution (25 μg / mL) and 20 μL of the vitamin C solution (25 μg / mL) were added, along with 180 μL of ABTS working solution. The absorbance value of the sample solution was recorded as Sample A, and the absorbance value of the vitamin C solution was recorded as Sample A. Vc The model group was set up (20 μL distilled water and 180 μL ABTS working solution), and the absorbance values ​​were denoted as Model A, A... Vc A blank control group (20 μL distilled water and 180 μL PBS working solution) was set up, and the absorbance value was recorded as blank A. According to the formula, when the solution concentration was 25 μg / mL, the ABTS radical scavenging rate of the novel spirulina-derived pentapeptide LPELY solution was 96.65%, and the ABTS radical scavenging rate of the vitamin C solution was 100%.

[0059] Example 10:

[0060] The antioxidant properties of the novel spirulina-derived pentapeptide LPELY solution and vitamin C solution (50 μg / mL, Example 1) were compared and tested separately. The samples were measured as follows: 20 μL of the novel spirulina-derived pentapeptide LPELY solution (50 μg / mL) and 20 μL of the vitamin C solution (50 μg / mL) were added, along with 180 μL of ABTS working solution. The absorbance value of the sample solution was denoted as Sample A, and the absorbance value of the vitamin C solution was denoted as AVc. A model group (20 μL distilled water and 180 μL ABTS working solution) was set up, with absorbance values ​​denoted as Model A and AVc. A blank group (20 μL distilled water and 180 μL PBS working solution) was set up, with absorbance values ​​denoted as Blank A. According to the formula, when the solution concentration was 50 μg / mL, the ABTS free radical scavenging rate of the novel spirulina-derived pentapeptide LPELY solution was 98.87%, and the ABTS free radical scavenging rate of the vitamin C solution was 100%.

[0061] Example 11:

[0062] The antioxidant properties of the novel spirulina-derived pentapeptide LPELY solution and vitamin C solution (100 μg / mL, Example 1) were compared and tested separately. The samples were measured as follows: 20 μL of a 100 μg / mL sample solution and 20 μL of a 100 μg / mL vitamin C solution were added, along with 180 μL of ABTS working solution. The absorbance value of the novel spirulina-derived pentapeptide LPELY solution was recorded as Sample A, and the absorbance value of the vitamin C solution was recorded as AVc. A model group (20 μL distilled water and 180 μL ABTS working solution) was set up, with absorbance values ​​recorded as Model A and AVc. A blank group (20 μL distilled water and 180 μL PBS working solution) was set up, with absorbance values ​​recorded as Blank A. According to the formula, when the solution concentration is 100 μg / mL, the ABTS radical scavenging rate of the novel spirulina-derived pentapeptide LPELY solution is 99.75%, and the ABTS radical scavenging rate of the vitamin C solution is 100%.

[0063] It should be noted that:

[0064] Determination of pancreatic lipase inhibition rate:

[0065] Add 50 μL (6 mg / mL) of the novel spirulina-derived pentapeptide (LPELY), 25 μL (5 mM) of 4-nitrophenylbutyrate, and 25 μL (1.25 mg / mL) of pancreatic lipase to a 96-well plate. Adjust the volume to 150 μL with PBS, incubate at 37°C for 45 minutes, and measure the absorbance at 405 nm. The inhibitory activity is calculated using the following formula:

[0066]

[0067] Where: A: No sample added, enzyme added; B: No sample added, enzyme added; C: Sample added, enzyme added; D: Sample added, enzyme not added.

[0068] Evaluation of the antioxidant activity of peptides:

[0069] ABTS free radical scavenging activity assay: A 5 mmol / L ABTS solution was prepared using PBS (pH 7.4), and excess MnO2 was added to prepare free radicals. The solution was incubated at 30°C for 12 h, followed by centrifugation at 4500 r / min for 10 min. The supernatant was collected and filtered through a 0.2 μm nylon membrane. The filtrate was stored at -20°C for later use as ABTS. + Stock solution. Before use in testing, ABTS needs to be dissolved in PBS solution. + Dilute the stock solution to the required concentration. Assess the scavenging activity of ABTS radicals using a 96-well plate. Add 20 μL of water and 180 μL of ABTS solution, perform a full-wavelength scan, and determine the maximum absorption wavelength as 736 nm (ELISA reader program: oscillation 30 s, measurement wavelength range 500-800 nm).

[0070] Vitamin C (Vc) was used as a positive control. When testing the ABTS free radical scavenging activity of the samples and vitamin C, the absorbance of the system was adjusted to 0.70 ± 0.02 to obtain the working solution. Sample determination: 20 μL of sample at different concentrations (25-800 μg / mL) or Vc (1-100 μg / mL) and 180 μL of ABTS working solution were added. The absorbance value of the sample was recorded as A. 样品 The model group was set up (20 μL distilled water and 180 μL ABTS working solution), and the absorbance value was denoted as A. 模型 Set up a blank control group (20 μL distilled water and 180 μL PBS working solution), and record the absorbance value as A. 空白 .

[0071] The scavenging rate of the sample for ABTS radicals was calculated using the following formula:

[0072] ABTS free radical scavenging rate = (A 模型 -A 样品 ) / (A 模型 -A 空白 )

[0073] The results of Examples 2-11 above show that the novel spirulina-derived pentapeptide LPELY of the present invention has high ABTS free radical scavenging activity. When the sample concentration is 50 μg, the free radical scavenging rate is 98.87%, and when the sample concentration is 100 μg, the free radical scavenging rate is as high as 99.75%, indicating that the novel spirulina-derived pentapeptide LPELY of the present invention has high antioxidant activity.

[0074] In addition, from Figure 3 It can also be seen that the novel spirulina-derived pentapeptide LPELY of this application has a high pancreatic lipase inhibition rate. Figure 4 The results show that the novel spirulina-derived pentapeptide LPELY has an excellent inhibition rate against free radicals.

[0075] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0076] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. A spirulina-derived pentapeptide, LPELY, characterized in that, The spirulina-derived pentapeptide LPELY has a molecular weight of 633.72 Da and a sequence of Leu-Pro-Glu-Leu-Tyr. Its chemical structure is as follows:

2. A method for purifying LPELY, a pentapeptide derived from Spirulina, characterized in that, The purification method is used to purify the Spirulina-derived pentapeptide LPELY as described in claim 1, and the purification method includes the following steps: The spirulina-derived pentapeptide LPELY to be purified was loaded onto the chromatographic column; gradient elution was performed using mobile phase A and mobile phase B as eluents, where mobile phase A was an aqueous solution containing 0.1% trifluoroacetic acid by volume, and mobile phase B was an acetonitrile solution containing 0.1% trifluoroacetic acid by volume; the gradient elution program was as follows: the initial volume proportion of mobile phase A was 10%, and from 0.01 min to 25 min after loading, the volume proportion of mobile phase A increased to 10%~70%, and from 25 min to 25.01 min, the volume proportion of mobile phase A increased to 100%, and the process was maintained at 100% until the operation was stopped, with a detection wavelength of 220 nm; the spirulina-derived pentapeptide LPELY solution of the target peak was collected.

3. The purification method according to claim 2, characterized in that, The chromatographic column is C. 18 The chromatographic column; and / or, mobile phase A is an aqueous solution containing 0.1% trifluoroacetic acid by volume, and mobile phase B is an acetonitrile solution containing 0.1% trifluoroacetic acid by volume; and / or, during gradient elution, the initial volume proportion of mobile phase A is 10%, and the volume proportion of mobile phase A increases to 70% within 0.01 min to 25 min after loading, and increases to 100% within 25 min to 25.01 min.

4. An application of LPELY, a pentapeptide derived from spirulina, characterized in that, The use of the spirulina-derived pentapeptide LPELY as described in any one of claims 1 to 3 in the preparation of lipid-lowering drug products.

5. An application of LPELY, a pentapeptide derived from spirulina, characterized in that, The application of the spirulina-derived pentapeptide LPELY as described in any one of claims 1 to 3 in the preparation of cosmetics.

6. An application of LPELY, a pentapeptide derived from spirulina, characterized in that, The application of the spirulina-derived pentapeptide LPELY as a food additive in food preparation, as described in any one of claims 1 to 3.

7. The application of the spirulina-derived pentapeptide LPELY according to claim 4, characterized in that, The spirulina-derived pentapeptide LPELY is used as a lipid-lowering and / or antioxidant active ingredient in the preparation of cosmetics or food.

8. A composition, characterized in that, The composition contains the spirulina-derived pentapeptide LPELY as described in claim 1.

9. The composition according to claim 8, characterized in that, The composition is one of a lipid-lowering drug composition, a cosmetic composition, and a food composition.

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