Instant solid beverage of multi-source composite peptide and enzymolysis preparation method of instant solid beverage
Through five-member complex protein source enzymatic lysis, six-enzyme synergistic dynamic enzymatic lysis and supercritical drying technology, problems such as single protein source, inaccurate enzymatic lysis and large drying particle size in multi-source peptide products were solved, and a multi-source complex peptide beverage with high amino acid coverage, rapid dissolution and stable were prepared.
Patent Information
- Application Number
- CN202510781146.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-06-12
- Publication Date
- 2025-09-05
AI Technical Summary
In the prior art, multi-source peptide products have problems such as single protein source, insufficient amino acid balance, inability to direct control of the molecular weight of the peptide segment during the enzymatic process, and large particle size and poor instant solubility in the drying process.
The five-member complex protein source enzymatic lysis, six-enzyme synergistic dynamic enzymatic lysis, nanofiltration staging purification and supercritical drying technology were used to prepare multi-source complex peptide instantly dissolved solid beverages. Through ultrasonic pretreatment, temperature-changing pH enzymatic lysis, nanofiltration staging purification and supercritical drying, the targeted preparation and functional enhancement of specific peptides are achieved.
The high amino acid coverage, good biological activity and excellent stability of multi-source composite peptide beverages are achieved. The product is quickly dissolved in cold water and has stable storage for a long time, forming a synergistic effect between functional components, significantly improving absorption efficiency and physiological regulation functions.
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Figure CN120585025A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the field of functional food processing technology, and specifically relates to an instant solid beverage of multi-source composite peptides and an enzymatic preparation method thereof. Background Art
[0002] In the field of functional foods, peptide products have attracted much attention due to their advantages such as easy absorption and high biological activity. However, existing technologies have the following significant drawbacks: Single protein source: Traditional multi-source peptides usually only contain the animal-plant-microorganism ternary system, and lack the application of new sustainable protein sources such as insect protein such as black soldier fly and algae protein such as spirulina, resulting in insufficient amino acid balance and essential amino acid coverage generally below 90%.
[0003] The enzymatic hydrolysis process is crude: conventional enzymatic hydrolysis technology uses fixed pH and temperature, which cannot directionally control the molecular weight of peptides. The proportion of target peptides of 500-1000Da is mostly less than 80%, and there is a lack of modification technology for peptide functional groups, making it difficult to give them specific functions such as antioxidant and immune regulation.
[0004] Backward processing technology: Traditional spray drying is commonly used in the drying process, resulting in large particle size, between 50-100μm, strong hygroscopicity, cold water dissolution time exceeding 30 seconds, and easy precipitation and stratification after standing. Summary of the Invention
[0005] To address the above problems, the present invention has constructed a breakthrough instant peptide beverage preparation system through innovative technologies such as five-element protein source compounding, six-enzyme coordinated dynamic enzymatic hydrolysis, nanofiltration graded purification and supercritical drying.
[0006] In order to solve the above technical problems, the technical solution adopted by the present invention is: A multi-source composite peptide instant solid beverage comprises the following components in percentage by weight: 20%-40% multi-source composite peptide, 30%-50% functional excipient, 5%-15% flavoring, and 5%-10% stabilizer; the multi-source composite peptide is prepared by enzymatic hydrolysis of five composite protein sources, the five composite protein sources including animal protein, plant protein, microbial protein, insect protein, and algae protein; the multi-source composite peptide comprises peptide segments with a molecular weight of 500-1000 Da accounting for ≥90%, and is modified to form active peptides with specific functional groups.
[0007] Furthermore, the animal protein includes one or more of mammalian protein, insect protein, and marine animal protein; the plant protein includes terrestrial plant protein and algae protein; and the microbial protein includes fermentation protein and fungal protein.
[0008] Furthermore, the specific functional groups include γ-glutamyl bonds, glycosylation sites or seleno groups, which are achieved by enzymatic modification or chemical modification.
[0009] A method for preparing an instant solid beverage by enzymatic hydrolysis of a multi-source composite peptide comprises the following steps: S1. The five-element complex protein source is pretreated to form a protein solution, which is first subjected to physical pretreatment; S2. A composite enzymatic hydrolysis system is added to the protein solution, and the enzymatic hydrolysis reaction is performed by dynamically controlling the reaction conditions; S3. Inactivate the enzyme, filter, and fractionate the hydrolyzate to separate peptides of different molecular weight ranges; S4. The peptide segment is mixed with auxiliary materials and then dried to prepare an instant solid beverage.
[0010] Furthermore, the physical pretreatment is ultrasonic pretreatment, and the treatment parameters are: frequency 20-40 kHz, power 0.5-1 kW, and treatment time 10-20 minutes.
[0011] Furthermore, the complex enzymatic hydrolysis system includes a main enzyme system and a coenzyme system: The main enzyme system includes neutral protease, alkaline protease and acidic protease; The coenzyme system includes an enzyme for decomposing microbial cell walls, an exonuclease for controlling peptide chain terminal groups, and a functional enzyme for peptide chain modification.
[0012] Furthermore, the neutral protease is Alcalase, the alkaline protease is Protex 6L, the acidic protease is pepsin, the enzyme used to decompose the microbial cell wall is endo-β-1,3-glucanase, the exoenzyme that controls the terminal groups of the peptide chain is oligopeptide exonuclease, and the functional enzyme used for peptide chain modification is glutamine aminotransferase or glycosidase.
[0013] Furthermore, the dynamic control of the reaction conditions includes temperature-pH gradient control, with the reaction temperature at 50-55°C and pH 7.0-7.5 for the first 2 hours, and the temperature gradually increasing to 55-60°C and the pH adjusted to 6.0-6.5 for the next 2-4 hours.
[0014] Furthermore, the fractional purification was performed by nanofiltration membrane separation with a molecular weight cut-off of 300Da, and low molecular weight peptides of <500Da and high molecular weight peptides of 500-1000Da were separated and used for the preparation of different functional components respectively.
[0015] Furthermore, the drying step adopts supercritical fluid drying or freeze drying, wherein the supercritical fluid drying pressure is 10-20 MPa and the temperature is 35-45° C. The particle size distribution of the prepared solid beverage particles is 10-20 μm, and a nano-scale hydrophilic coating is formed on the surface.
[0016] Compared with the prior art, the present invention has the following beneficial effects: The present invention provides a multi-source composite peptide instant solid beverage and its enzymatic hydrolysis preparation method. The beverage is mainly composed of multi-source composite peptides obtained by enzymatic hydrolysis of five composite protein sources including animals, plants, microorganisms, insects and algae, and is compounded with functional excipients, flavorings and stabilizers. During the preparation process, ultrasonic pretreatment is used to destroy the protein structure, and a composite enzyme system is used for dynamic enzymatic hydrolysis at variable temperature and pH. Nanofiltration graded purification and supercritical drying technology are combined to achieve the directional preparation and functional enhancement of the target peptide segment. This solution solves the problems of traditional peptide beverages such as single protein source, low activity and poor solubility through the synergy of multiple protein sources, precise enzymatic hydrolysis control and advanced processing technology. The resulting product has a comprehensive amino acid composition, high biological activity and excellent stability. A synergistic effect is formed between the functional ingredients, significantly improving the absorption efficiency and physiological regulation function. BRIEF DESCRIPTION OF THE DRAWINGS
[0017] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.
[0018] Figure 1 The figure is a flow chart of the enzymatic preparation method of the present invention. DETAILED DESCRIPTION
[0019] The following is a clear and complete description of the technical solutions in 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 embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.
[0020] The present invention provides an instant solid beverage of multi-source composite peptides and an enzymatic hydrolysis preparation method thereof, which specifically includes the following component proportions and a preparation method: A multi-source composite peptide instant solid beverage comprising the following components in percentage by weight: Multi-source complex peptide 20%-40% Functional excipients 30%-50%, including oligofructose, oligogalactose, etc. Flavoring 5%-15%, including sucrose, sucralose, etc. Stabilizer 5%-10%, including sodium carboxymethyl cellulose, xanthan gum, etc. Core innovation: Five-element complex protein source The multi-source composite peptide is prepared by enzymatic hydrolysis of five composite protein sources, including: Animal protein: This includes mammalian protein such as milk and beef, insect protein such as black soldier fly larvae, and marine animal protein such as Antarctic krill, providing essential amino acids and sulfur-containing amino acids; Plant protein: including terrestrial plant protein such as soy, quinoa and algae protein such as spirulina and chlorella, which are rich in dietary fiber and phytosterols; Microbial protein: Contains fermented protein such as fermented soybean meal and fungal protein such as Cordyceps militaris mycelium, providing immune active ingredients such as β-glucan.
[0021] Functional enhancement features Target peptide segment control: 500-1000Da peptide segment accounts for ≥90%, with both good absorption and biological activity; Functional group modification: The peptides are endowed with new functions such as antioxidant and anti-inflammatory properties by forming γ-glutamyl bonds through transglutaminase (TG), introducing glycosylation sites through glycosidase, or adding selenomethionine to form seleno groups.
[0022] 2. Preparation Method Step 1: Pretreatment of five-element protein source Animal protein, plant protein, microbial protein, insect protein, and algae protein are individually pulverized through an 80-100 mesh sieve and mixed in a ratio of 30%-50% animal protein, 30%-50% plant protein, 10%-20% microbial protein, 5%-15% insect protein, and 5%-15% algae protein. Deionized water is then added to create a 5%-20% (w / v) protein solution. Innovative pretreatment: Ultrasonic treatment is used at a frequency of 20-40kHz, a power of 0.5-1kW, and a treatment time of 10-20 minutes. This cavitation effect disrupts the protein's higher-order structure, increasing the exposure of enzymatic sites by over 40%.
[0023] Step 2: Six-enzyme synergistic dynamic enzymolysis Composite enzymatic hydrolysis system, total enzyme content accounts for 0.1%-1.0% of protein content (w / w): The main enzyme system has a mass ratio of 2:1:1: neutral protease such as Alcalase, alkaline protease such as Protex 6L, and acidic protease such as pepsin, which synergistically hydrolyze different protein structures; Coenzyme system: endo-β-1,3-glucanase, used to decompose microbial cell walls; oligopeptidase, used to control the C / N-terminal groups of peptide chains; TG enzyme or glycosidase, used to modify functional groups.
[0024] Dynamic control conditions: First 2 hours: The temperature is controlled at 50-55°C, the pH is maintained at 7.0-7.5, and the main enzyme system quickly opens the tertiary structure of the protein; Last 2-4 hours: Gradient temperature is increased to 55-60°C, and pH is adjusted to 6.0-6.5. The coenzyme system accurately cuts the peptide chain and modifies the functional groups, so that the proportion of 500-1000Da peptide segments can reach 92%.
[0025] Step 3: Nanofiltration fractionation purification Separation by nanofiltration membrane with a molecular weight cut-off of 300Da: Low molecular weight peptides <500Da, enriched with antioxidant functions, such as peptides containing seleno groups; High molecular weight peptides of 500-1000Da enhance immune regulatory functions, such as peptides containing γ-glutamyl bonds, to achieve targeted separation of functional components.
[0026] Step 4: Supercritical Fluid Drying Supercritical CO2 spray drying is used at a pressure of 10-20MPa and a temperature of 35-45°C to form a nano-scale hydrophilic coating on the surface of the particles. The particle size distribution is 10-20μm, the cold water dissolution time is ≤3 seconds, and there is no precipitation after standing for 24 hours. The hygroscopicity is reduced by 30% compared with the traditional process.
[0027] The following are specific embodiments of the present invention: Example 1: Antioxidant beverage containing black soldier fly protein Raw material composition (weight percentage) Multi-source complex peptide 30%: milk protein 20%, black soldier fly larvae protein 15%, quinoa protein 20%, fermented soybean meal protein 25%, spirulina protein 20%. After enzymatic hydrolysis, the 500-1000Da peptide segment accounts for 92%, containing 0.8% γ-glutamyl bond modified peptides; Functional excipients 40%, including oligofructose 25% and inulin 15%; Flavoring 12%, including sucrose 8%, sucralose 4%; Stabilizer 8%, including sodium carboxymethyl cellulose 5%, xanthan gum 3%.
[0028] Preparation steps Protein pretreatment: The five proteins were mixed and crushed through a 100-mesh sieve to prepare a 10% (w / v) solution, which was then ultrasonically treated at 30 kHz and 1 kW for 15 minutes. Enzymatic hydrolysis: Add complex enzymes with a mass ratio of Alcalase: Protex 6L: pepsin: glucanase: exoenzyme: TG enzyme = 2:1:1:0.5:0.5:0.8. Control the temperature at 50°C and pH 7.2 for the first 2 hours. Then raise the temperature to 55°C and adjust the pH to 6.3 for the next 3 hours. The total enzymatic hydrolysis time is 5 hours. Nanofiltration separation: collect 500-1000Da peptides and mix them with excipients; Supercritical drying: Drying under 15 MPa and 40°C to obtain a light yellow powder with a particle size of 15 μm.
[0029] Performance Testing Dissolving time: 2.8 seconds in cold water at 25°C. Stability: No precipitation after storage at 4°C for 30 days, turbidity <5 NTU after reconstitution; Biological activity: DPPH scavenging rate IC50 = 0.22mg / ml, 45% higher than the traditional process; Caco-2 cell absorption rate 68%.
[0030] Example 2: Immunomodulatory beverage containing Cordyceps militaris protein Raw material composition (weight percentage) Multi-source complex peptide 35%: egg protein 25%, Antarctic krill protein 15%, chia seed protein 20%, Cordyceps militaris mycelium protein 20%, Chlorella protein 20%, after enzymatic hydrolysis, 500-1000Da peptides account for 91%, containing 1.2% glycosylated modified peptides; Functional excipients 35%, including galacto-oligosaccharides 20% and resistant dextrin 15%; Flavoring 10%, including fructose 8%, aspartame 2%; Stabilizer 10%, including sodium alginate 6% and gelatin 4%.
[0031] Preparation differences Enzymatic hydrolysis system: The coenzyme system uses glycosidase to replace TG enzyme to modify and form β-1,6-glycopeptide bonds; Drying process: freeze drying is used, with a pre-freezing temperature of -35°C, a vacuum degree of 5Pa, and a drying time of 48 hours to retain the activity of thermosensitive glycosylated peptides.
[0032] Performance Testing Immune activity: Promoted the proliferation rate of mouse spleen lymphocytes by 45%, an increase of 38% compared with the control group; Solubility: 30g dissolves in 100ml cold water without turbidity; Storage characteristics: When placed in an environment with a relative humidity of 75% for 30 days, the moisture absorption rate is less than 5%.
[0033] The present invention provides a multi-source composite peptide instant solid beverage and an enzymatic hydrolysis preparation method thereof. The beverage contains 20%-40% of multi-source composite peptides prepared by enzymatic hydrolysis of five composite protein sources including animals, plants, microorganisms, insects and algae, as well as functional excipients, flavorings and stabilizers, wherein the 500-1000Da peptide segment accounts for ≥90%, and is enzymatically modified to form active peptides containing specific functional groups; the preparation method comprises ultrasonic pretreatment of the five-element protein source, using a composite enzymatic hydrolysis system comprising a main enzyme system and a coenzyme system to perform a temperature-variable pH dynamic enzymatic hydrolysis reaction, inactivating the enzyme in the enzymatic hydrolysis solution, filtering and then grading and purifying the enzymatic hydrolysis solution through a nanofiltration membrane, and finally mixing the peptide segment with the excipient and spray drying or freeze-drying the peptide segment with supercritical CO2. Through the combination of five protein sources, precise enzymatic hydrolysis control and advanced processing technology, this solution achieves 100% coverage of essential amino acids in the product, a high proportion of target peptides and significantly improved biological activity, and a 90% increase in solubility compared to traditional processes, achieving cold water dissolution within 3 seconds and no precipitation for 24 hours. Functional excipients and modified peptides form a synergistic network, effectively enhancing intestinal mucosal absorption and maintaining the activity of antioxidant enzymes in the body, solving problems such as single protein source, extensive enzymatic hydrolysis and backward processing in existing technologies.
[0034] In addition, it should be understood that although this specification is described in terms of implementation methods, not every implementation method contains only one independent technical solution. This narrative method of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment can also be appropriately combined to form other implementation methods that can be understood by those skilled in the art.
Claims
1. A multi-source composite peptide instant solid beverage, characterized in that: The invention comprises the following components in weight percentage: 20%-40% multi-source composite peptides, 30%-50% functional excipients, 5%-15% flavorings, and 5%-10% stabilizers; the multi-source composite peptides are prepared by enzymatic hydrolysis of five composite protein sources, and the five composite protein sources include animal protein, plant protein, microbial protein, insect protein, and algae protein; the peptide segments with a molecular weight of 500-1000 Da account for ≥90% of the multi-source composite peptides, and are modified to form active peptides with specific functional groups.
2. The instant solid beverage according to claim 1, characterized in that The animal protein includes one or more of mammalian protein, insect protein, and marine animal protein; the plant protein includes terrestrial plant protein and algae protein; and the microbial protein includes fermentation protein and fungal protein.
3. The instant solid beverage according to claim 1, characterized in that The specific functional groups include γ-glutamyl bonds, glycosylation sites or seleno groups, which are achieved by enzymatic modification or chemical modification.
4. The enzymatic hydrolysis method for preparing a multi-source composite peptide instant solid beverage according to any one of claims 1 to 3, characterized in that: The following steps are involved: S1. The five-element complex protein source is pretreated to form a protein solution, which is first subjected to physical pretreatment; S2. A composite enzymatic hydrolysis system is added to the protein solution, and the enzymatic hydrolysis reaction is performed by dynamically controlling the reaction conditions; S3. Inactivate the enzyme, filter, and fractionate the hydrolyzate to separate peptides of different molecular weight ranges; S4. The peptide segment is mixed with auxiliary materials and then dried to prepare an instant solid beverage.
5. The preparation method according to claim 4, characterized in that The physical pretreatment is ultrasonic pretreatment, and the processing parameters are: frequency 20-40kHz, power 0.5-1kW, and processing time 10-20 minutes.
6. The preparation method according to claim 4, characterized in that The complex enzymatic hydrolysis system includes the main enzyme system and the coenzyme system: The main enzyme system includes neutral protease, alkaline protease and acidic protease; The coenzyme system includes an enzyme for decomposing microbial cell walls, an exonuclease for controlling peptide chain terminal groups, and a functional enzyme for peptide chain modification.
7. The preparation method according to claim 6, characterized in that The neutral protease is Alcalase, the alkaline protease is Protex 6L, the acidic protease is pepsin, the enzyme used to decompose the microbial cell wall is endo-β-1,3-glucanase, the exoenzyme for controlling the terminal groups of the peptide chain is oligopeptide exoenase, and the functional enzyme used for peptide chain modification is glutamine aminotransferase or glycosidase.
8. The preparation method according to claim 4, characterized in that Dynamically controlled reaction conditions include variable temperature-variable pH gradient control, with the reaction temperature at 50-55°C and pH 7.0-7.5 for the first 2 hours, and then the temperature gradually increased to 55-60°C and the pH adjusted to 6.0-6.5 for the next 2-4 hours.
9. The preparation method according to claim 4, characterized in that The fractionation and purification were performed by nanofiltration membrane separation with a molecular weight cut-off of 300Da, and low molecular weight peptides of <500Da and high molecular weight peptides of 500-1000Da were separated and used for the preparation of different functional components.
10. The preparation method according to claim 4, characterized in that The drying step adopts supercritical fluid drying or freeze drying, wherein the supercritical fluid drying pressure is 10-20MPa and the temperature is 35-45°C. The particle size distribution of the prepared solid beverage particles is 10-20μm, and a nano-scale hydrophilic coating is formed on the surface.