Application of Spirulina Phycocyanin Peptides in the Preparation of Anti-aging Products and Spirulina Phycocyanin Peptide Beer
By adding spirulina phycocyanin peptides during the beer brewing process, especially by adding 6-8‰ of the dry weight of malt before the end of the saccharification step, spirulina phycocyanin peptide beer is prepared, which solves the problem that beer products are difficult to meet nutritional and health care functions, and achieves the anti-aging effect of beer and the effect of rich and white beer foam.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- QILU UNIVERSITY OF TECHNOLOGY (SHANDONG ACADEMY OF SCIENCES)
- Filing Date
- 2025-11-21
- Publication Date
- 2026-06-30
AI Technical Summary
Existing beer products are unable to meet consumers' diverse needs for nutritional and health benefits, and the application of spirulina phycocyanin peptides in beer brewing has not been widely studied.
Spirulina phycocyanin peptides are added during the beer brewing process, especially 6-8‰ of the dry weight of malt phycocyanin peptides added 5-8 minutes before the end of the saccharification step. Spirulina phycocyanin peptides are then separated and purified by enzymatic hydrolysis, ultrafiltration and high performance liquid chromatography to prepare spirulina phycocyanin peptide beer, which promotes the synthesis of type I collagen and has anti-aging effects.
The prepared spirulina phycocyanin peptide beer exerts a synergistic anti-aging effect during fermentation. The beer has rich and white foam, and its alcohol content and total acid content meet national standards. It also has excellent foam retention and good economic value and application prospects.
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Abstract
Description
Technical Field
[0001] This invention belongs to the field of protein peptide application and beer brewing technology, specifically relating to the application of spirulina phycocyanin peptide in the preparation of anti-aging products and spirulina phycocyanin peptide beer. Background Technology
[0002] With consumers becoming increasingly health-conscious, traditional industrial beer can no longer meet the market's diversified demands for nutritional and health benefits. Functional beer, with its unique nutritional value and health effects, is becoming a new trend in the industry. Studies have shown that functional beer not only retains the taste characteristics of traditional beer, but also endows it with antioxidant, anti-inflammatory, and immune-regulating health functions by adding functional ingredients (such as active peptides and plant extracts). This product design, combining drinkability and functionality, aligns with current consumers' pursuit of the "medicine and food homology" concept, indicating huge market potential.
[0003] Spirulina is rich in protein and low in lipids, making spirulina protein an excellent source of high-quality bioactive peptides. Its various functional properties have been extensively studied, including antioxidant, anti-aging, iron chelation, anti-atherosclerotic, anti-inflammatory, and antihypertensive activities. Spirulina phycocyanin peptides are widely used in pharmaceuticals, food, and cosmetics due to their excellent biocompatibility, easy absorption, and relatively safe characteristics. However, to date, few studies have explored the application of spirulina-derived peptides in beer brewing processes. Summary of the Invention
[0004] To address the problems existing in the prior art, this invention provides an application of spirulina phycocyanin peptide in the preparation of anti-aging products and spirulina phycocyanin peptide beer. Spirulina phycocyanin peptide has good anti-aging effects and can be used as an additive ingredient in functional beer, showing good application prospects in the beer industry.
[0005] This invention is achieved through the following technical solution:
[0006] In a first aspect, the present invention provides the application of spirulina phycocyanin peptide in the preparation of anti-aging products, wherein the amino acid sequence of the spirulina phycocyanin peptide is shown in SEQ ID NO.1.
[0007] Furthermore, the anti-aging product is spirulina phycocyanin peptide beer.
[0008] In a second aspect, the present invention provides a spirulina phycocyanin peptide beer, wherein spirulina phycocyanin peptide is added to the spirulina phycocyanin peptide beer.
[0009] Furthermore, the preparation process of the spirulina phycocyanin peptide beer includes malt crushing, saccharification, filtration, boiling, vortex sedimentation, and fermentation maturation steps.
[0010] Furthermore, spirulina phycocyanin peptides are added 5-8 minutes before the end of the saccharification step.
[0011] Furthermore, the amount of Spirulina phycocyanin peptide added is 6-8‰ of the dry weight of malt.
[0012] Furthermore, the spirulina phycocyanin peptides are extracted from Spirulina platensis or chemically synthesized. By optimizing the enzymatic hydrolysis conditions, enzymatic hydrolysis products with high activity are screened, and then separated and purified by ultrafiltration and high performance liquid chromatography (HPLC). The active peptide components of the phycocyanin source are then analyzed by LC-MS / MS, and the physicochemical properties, potential biological activities, toxicity, etc. of the peptides are predicted by bioinformatics methods, from which bioactive peptides are screened.
[0013] Compared with the prior art, the beneficial effects achieved by the present invention are as follows:
[0014] This invention creatively discovers that spirulina phycocyanin peptide (MGHP) can promote the synthesis of type I collagen and has anti-aging effects. By preparing it into functional beer, its anti-aging effect can be further enhanced in conjunction with the beer fermentation process, expanding the application of small molecule peptides in the food field, and has good application prospects and excellent economic value. Detailed Implementation
[0015] The present invention is further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods not specifically described in the following examples are generally performed under conventional conditions or as recommended by the manufacturer.
[0016] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of skill in the art. All reagents and materials used in this invention are readily available through conventional means, and unless otherwise specified, they shall be used in accordance with conventional methods in the art or as per the product instructions.
[0017] Example 1
[0018] Spirulina phycocyanin peptide was obtained according to the method disclosed in patent CN119350478B, and the amino acid sequence (SEQ ID NO.1) of the spirulina phycocyanin peptide is MGHP (Met-Gly-His-Pro).
[0019] Example 2
[0020] The preparation of spirulina phycocyanin peptide beer with anti-aging effects includes the following steps:
[0021] (1) Raw material crushing
[0022] Add water (20wt%) to the malt, stir thoroughly, and then grind it in a grinder to maximize the crushing of the malt endosperm. The standard for crushing is to break the bran but not crush it. This will form a layer of lees during filtration, which is beneficial for the filtration of wort.
[0023] (2) Saccharification
[0024] Barley malt and wheat malt were crushed at a ratio of 3:2 (dry weight of malt) and fed into the water at a ratio of 1:3.5. The water temperature was raised to 50°C before feeding. To ensure complete protein decomposition, the temperature was maintained for 30 minutes, and then the temperature was slowly raised to 68°C and maintained for 60 minutes to allow β-amylase to hydrolyze the protein under suitable conditions, which was intended to form fermentable sugars. Finally, the temperature was raised to 78°C. Five minutes before the end of saccharification, 6‰ (dry weight of malt) of spirulina phycocyanin peptide (MGHP) was added. A small amount of wort was then tested for iodine. If there was no obvious discoloration, the saccharification process was considered complete.
[0025] (3) Filtering
[0026] After saccharification, the wort is filtered. The wort is then poured into a filter tank and filtered using the filter layer naturally formed by the tank and the wasps. If the filtration process is too slow in the later stages, it can be adjusted by manually stirring (in the bottle) or rotating the tiller (in the pot). When filtration is almost complete, the wasps are washed with 78°C water, and the sugar content of the wort is tested. Throughout the process, the filtered wort must be clear and transparent. Finally, the filtered wort is poured into a boiling kettle.
[0027] (4) Boil
[0028] After filtering the wort, boil it in a boiling pot and keep it boiling for 70 minutes. Add hops three times during the boiling process, starting from the moment the wort boils. Add 0.1‰ bitter hops after boiling for 10 minutes, add 0.3‰ bitter hops after boiling for 30 minutes, and add 0.2‰ aromatic hops 10 minutes before the end of boiling.
[0029] (5) vortex precipitation
[0030] After boiling, pour the wort into a vortex settling tank and let it stand for 20 minutes.
[0031] (6) Fermentation and ripening
[0032] After swirling and settling, the hop mash and hot coagulated material are discharged. The clear wort is cooled via a plate heat exchanger and pumped into the fermentation tank. The inlet temperature is controlled at 18-22°C, and oxygen is introduced to create an environment conducive to yeast initiation and growth. Yeast cells are then inoculated under aseptic conditions. To ensure good yeast growth and metabolic activity, the inoculation concentration is 0.8 × 10⁻⁶. 7 / mL; After the yeast is inoculated, the wort enters the main fermentation process (fermentation temperature controlled at 18-22℃). During the main fermentation process, the yeast performs aerobic respiration and consumes the sugar in the wort. Because the yeast multiplies vigorously at this time, it is necessary to frequently check the sugar content of the fermentation liquid. When the sugar content drops to 4°P, the tank is sealed (controlling the opening of the exhaust valve on the top of the tank to keep the tank pressure rising to the required value), and the main fermentation period ends.
[0033] After the fermentation tank is sealed, the post-fermentation process begins. During this time, the yeast undergoes anaerobic respiration, producing large amounts of carbon dioxide and ethanol, causing a rise in pressure within the fermentation tank. When the pressure reaches 0.14 MPa, diacetyl is reduced, a process that typically takes about a week. The diacetyl content in the fermentation broth is monitored daily. When the diacetyl content falls below 0.08 mg / L, the diacetyl reduction process is terminated, and the fermentation tank temperature is lowered to 0°C, maintaining the pressure at approximately 0.13 MPa. During fermentation, if excessive yeast sludge accumulates at the bottom of the tank, it must be drained appropriately to avoid affecting the beer's taste. After the fermentation temperature reaches 0°C, it is maintained for 7 days to enter the beer maturation period, aiming to stabilize the beer's body. The pressure within the fermentation tank is monitored, and to ensure a crisp and refreshing beer, food-grade CO2 gas is introduced when the pressure is insufficient.
[0034] Example 3
[0035] Compared with Example 2, in Example 3, the amount of Spirulina phycocyanin peptide (MGHP) added in step (2) is 7‰ of the dry weight of malt, and the rest of the operation is the same as in Example 2.
[0036] Example 4
[0037] Compared with Example 2, in Example 4, the amount of Spirulina phycocyanin peptide (MGHP) added in step (2) is 8‰ of the dry weight of malt, and the rest of the operation is the same as in Example 2.
[0038] Comparative Example 1
[0039] Compared with Example 2, the amount of Spirulina phycocyanin peptide (MGHP) added in step (2) of Comparative Example 1 was 2‰ of the dry weight of malt, and the rest of the operation was the same as in Example 2.
[0040] Comparative Example 2
[0041] Compared with Example 2, in Comparative Example 2, the amount of Spirulina phycocyanin peptide (MGHP) added in step (2) was 4‰ of the dry weight of malt, and the rest of the operation was the same as in Example 2.
[0042] Comparative Example 3
[0043] Compared with Example 2, in Comparative Example 3, the amount of Spirulina phycocyanin peptide (MGHP) added in step (2) was 1% of the dry weight of malt, and the rest of the operation was the same as in Example 2.
[0044] Comparative Example 4
[0045] Compared with Example 2, in Comparative Example 4, step (2) of the saccharification process did not involve the addition of spirulina phycocyanin peptide (MGHP), and the remaining operations were the same as in Example 2.
[0046] Experimental Example 1
[0047] Determination of physicochemical properties of beer:
[0048] (1) Alcohol content test: determined according to the volumetric method in the density bottle method of GB / T4928-2008 "Beer Analysis Methods".
[0049] (2) Determination of diacetyl content: The diacetyl content was determined according to the method for determining diacetyl content in GB / T4928-2008 "Beer Analysis Methods"; diacetyl was distilled out by distillation, and then diacetyl reacted with o-phenylenediamine to generate 2,3-dimethylquinoxaline. The absorbance was measured at a wavelength of 335 nm to detect the diacetyl content.
[0050] (3) Turbidity determination: After degassing the beer without filtering, the turbidity of the beer is determined by a turbidity meter according to GB / T4928-2008 "Beer Analysis Methods".
[0051] (4) Color determination: The color of beer is determined according to the EBC colorimeter method in GB / T4928-2008 "Beer Analysis Methods". Pour the degassed beer into a cuvette and compare it with the standard color wheel. When the color tone is consistent, read the color of the beer in EBC.
[0052] (5) Foam measurement: Observe the fineness, color and degree of foam clinging to the cup with your eyes.
[0053] (6) Effervescence determination: Place the beer sample in a water bath at about 20°C and keep it at a constant temperature for 30 minutes. Clean the effervescence cup thoroughly and set aside. Then, test it according to the stopwatch method in the national standard GB / T4928-2008 "Beer Analysis Methods".
[0054] (7) pH value determination of beer: Clean the beaker and electrode with distilled water, then use absorbent paper to dry the water on the electrode, then use standard solution to calibrate the pH meter. After calibration, clean the electrode with distilled water again, wipe the distilled water on the electrode dry, insert it into the test liquid, and wait for the data to display steadily before reading the value directly.
[0055] (8) Determination of total acidity in beer
[0056] First, prepare a standard sodium hydroxide solution (0.1 mol / L) and a phenolphthalein reagent (5 g / L). In a conical flask, add 100 mL of distilled water and boil for 2 min. Then add 10 mL of the sample, heat for another 1 min, turn off the heat, let stand for 5 min, rinse the flask with tap water, add 0.5 mL of phenolphthalein reagent, and titrate with 0.1 mol / L sodium hydroxide solution until a pale pink color is achieved. Record the volume of sodium hydroxide solution consumed.
[0057] Total acid = 10 × C × V;
[0058] C – Concentration of sodium hydroxide solution, mol / L;
[0059] V – The volume of sodium hydroxide solution consumed, in mL.
[0060] (9) The physicochemical properties of the beers prepared in Examples 2-4 and Comparative Examples 1-4 were tested, and the results are shown in Table 1 below:
[0061] Table 1 Comparison of Physicochemical Indicators of Beer
[0062]
[0063] As shown in Table 1, the physicochemical properties of beer are directly related to the entire brewing process and the amount of spirulina protein peptides added. The beer foam is rich, white and delicate, with good cling to the glass. The alcohol content, diacetyl content and total acid content all meet the national standard requirements. The alcohol content of the beer in Examples 2-4 is higher than that of Comparative Examples 1-4, and the foam retention of Examples 3 and 4 is better.
[0064] Experimental Example 2
[0065] Beer sensory evaluation
[0066] After the beer stabilized, a sensory evaluation was conducted on the finished beer by a 10-member judging panel (personnel trained in beer tasting from the School of Bioengineering, Qilu University of Technology (Shandong Academy of Sciences)). The beer was comprehensively evaluated based on color, clarity, foam, aroma, and taste. The detailed sensory evaluation scoring criteria are shown in Table 2, with a maximum score of 100 points, and the average score was taken. The beer sensory evaluation table is shown in Table 3.
[0067] Table 2 Sensory Evaluation Scoring Rules
[0068]
[0069] Table 3. Sensory Evaluation Table for Beer
[0070]
[0071] As shown in Table 3, the beers prepared in Examples 2-4 scored higher, at 93, 93, and 94 respectively. They were clear, bright in color, with fine white foam, harmonious aroma, rich fragrance, full-bodied and mellow taste, and a refreshing seaweed aroma. Comparative Examples 1-4 scored lower than Examples 2-4 in terms of aroma, taste, body richness, foam, and clarity.
[0072] Experimental Example 3
[0073] Experiment to promote the synthesis of type I collagen (collagen I)
[0074] The experiment used human skin fibroblasts (HSF) as a model cell line for research.
[0075] 1) Cell seeding: at 4 × 10 4 Human skin fibroblasts (HSF) were seeded into 24-well plates at a seeding density of cells / well and incubated overnight in an incubator (37°C, 5% CO2).
[0076] 2) Solution preparation: Prepare the sample solutions to be tested according to the test protocol table (Table 4);
[0077] 3) Drug administration: According to the test protocol in Table 4, when the cell deposition rate in the 24-well plate reaches 40%~60%, the sample solution to be tested is administered in groups, with 2 mL of sample added to each well and 3 replicates per group; after drug administration, the 6-well plate is placed in an incubator (37℃, 5% CO2) for 24 h.
[0078] 4) UVA irradiation: According to the experimental groups, the groups requiring UVA irradiation were subjected to 30J / cm² irradiation. 2 The sample was irradiated with UVA and then placed in an incubator (37℃, 5% CO2) for 24 hours.
[0079] 5) Sample collection: After culturing for 24 hours, collect the cell culture supernatant into EP tubes and freeze them at -80℃.
[0080] 6) ELISA assay: Perform the assay according to the ELISA kit's instruction manual;
[0081] 7) Statistical Analysis of Results: GraphPad Prism was used for plotting, and the results are expressed as averages. t-tests were used for comparisons between groups. 0.01 < pA value <0.05 is considered statistically significant. p A value <0.01 is considered highly significant.
[0082] Table 4 Test Plan Table
[0083] .
[0084] The results of collagen I content detection are shown in Table 5. Table 5 shows that compared with group BC, the collagen I content in group NC decreased significantly, indicating that the stimulation conditions in this test were effective. Compared with group NC, the collagen I content in group PC increased significantly, indicating that the positive control test was effective. Compared with group NC, Examples 2-4 and Comparative Examples 1-4 all promoted collagen I content. Examples 2-4 significantly increased collagen I content, with increases of 56.49%, 67.43%, and 73.69%, respectively. The promoting effect of Examples 2-4 was significantly higher than that of Comparative Examples 1-4, with Example 4 showing the highest increase rate. Therefore, this beer can significantly increase collagen I content, achieving a firming effect.
[0085] Table 5 Summary of Collagen I Content Detection Results
[0086]
[0087] (Note: ## indicates comparison with blank control,) p <0.01; ** indicates a difference compared to the negative control. p <0.01; * indicates that compared with the negative control, 0.01 < p <0.05).
[0088] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to the given embodiments, those skilled in the art can make modifications or equivalent substitutions to the technical solutions of the present invention as needed, without departing from the spirit and scope of the technical solutions of the present invention.
Claims
1. The application of Spirulina phycocyanin peptides in the preparation of anti-aging products, characterized in that, The amino acid sequence of the Spirulina phycocyanin peptide is shown in SEQ ID NO.1; The product in question is a cosmetic. The spirulina phycocyanin peptides described above can promote the synthesis of type I collagen.