American ginseng probiotic beverage and preparation method thereof

Abstract

The invention discloses a panax quinquefolius probiotic beverage and a preparation method thereof, and relates to the technical field of functional foods. According to the beverage, the combination of the American ginseng extract and specific probiotics is taken as a core, the survival rate of thalli is increased by adopting a microcapsule embedding technology, and the product performance is optimized by adding a stabilizer and a flavor regulator. The formula comprises 5-15% of a panax quinquefolius extract, 3-8% of compound probiotic freeze-dried powder (lactobacillus plantarum, lactobacillus salivarius and bifidobacterium lactis), 10-25% of inulin, 5-15% of whey protein, 10-30% of a microcapsule wall material (sodium alginate / chitosan or a WPI-KGM compound), and auxiliary materials. According to the preparation method, the active ingredients are efficiently reserved through the processes of gradient extraction, microcapsule embedding, low-temperature drying and the like. The product has the functions of resisting fatigue, regulating intestinal flora and enhancing immunity, the survival rate of probiotics in simulated gastric juice reaches 85% or above, and the shelf life stability is improved by 40%.

Description

Technical Field

[0001] The present invention relates to the technical field of functional foods, and particularly relates to an American ginseng probiotic drink and a preparation method thereof. Background Art

[0002] Currently, there are three major bottlenecks in American ginseng drinks: low retention rate of active ingredients, poor gastric acid tolerance of probiotics, and single function. The traditional extraction process uses high-temperature decoction, resulting in more than 40% isomerization loss of ginsenoside Rg1 and a polysaccharide yield of less than 7%. In terms of probiotic delivery, single sodium alginate embedding has a too high porosity (pore diameter > 100nm), and the viable bacteria survival rate is less than 20% after 2 hours in the gastric acid environment, severely restricting the functional effectiveness. Although existing combined products combine ginseng and probiotics, they do not construct a "component - flora - function" synergistic system: there is no two-way promotion between American ginseng polysaccharide and lactic acid bacteria metabolism, and the function only stays at the intestinal regulation level, and the potential of anti-fatigue and immune enhancement has not been fully explored. In addition, the single dosage form and short shelf life (the viable bacteria decay to less than 10 6 CFU / g after 6 months) further limit the industrial application. Summary of the Invention

[0003] The purpose of this part is to outline some aspects of the embodiments of the present invention and briefly introduce some preferred embodiments. Some simplifications or omissions may be made in this part, as well as in the abstract and title of the specification of this application, to avoid obscuring the purpose of this part, the abstract, and the title, and such simplifications or omissions shall not be used to limit the scope of the present invention.

[0004] In view of the problems existing in the above-mentioned prior art, the inventors of the present invention have proposed the present invention.

[0005] Therefore, the purpose of the present invention is to overcome the deficiencies in the prior art and provide an American ginseng probiotic drink and a preparation method thereof.

[0006] To solve the above technical problems, the present invention provides the following technical solution: an American ginseng probiotic drink, comprising the following components in weight percentages: 5 - 15% of American ginseng extract, 3 - 8% of compound probiotic freeze-dried powder, 10 - 25% of inulin, 5 - 15% of whey protein, 10 - 30% of microcapsule wall material, 0.5 - 2% of N-octenyl succinic anhydride, and the balance being purified water or maltodextrin.

[0007] As a preferred embodiment of the American ginseng probiotic drink of the present invention, wherein: the compound probiotic comprises Lactobacillus plantarum GP258, Lactobacillus salivarius LS45, and Bifidobacterium lactis BB12, and the mass ratio is (4 - 6):(2 - 3):(1 - 2).

[0008] As a preferred embodiment of the American ginseng probiotic drink of the present invention, wherein: the microcapsule wall material is selected from any one of the following:

[0009] a) Mulberry silk gum - sodium alginate - chitosan complex (SG:SA:CS = 4:6:2);

[0010] b) Whey protein - konjac glucomannan complex (WPI:KGM = 3:1).

[0011] As a preferred embodiment of the American ginseng probiotic drink of the present invention, wherein:.

[0012] As a preferred embodiment of the American ginseng probiotic drink of the present invention, wherein: the preparation method of the American ginseng extract includes a gradient separation step of subcritical water extraction of saponins and ultrasonic - assisted extraction of polysaccharides.

[0013] A preferred embodiment of a preparation method of an American ginseng probiotic drink, wherein: it includes the following steps:

[0014] (1) Preparation of American ginseng extract;

[0015] (2) Probiotic microencapsulation;

[0016] (3) Component mixing and sterilization;

[0017] (4) Filling or drying and forming.

[0018] As a preferred embodiment of a preparation method of an American ginseng probiotic drink of the present invention, wherein: in step (2), the W / O / W double - emulsion method is adopted, the inner aqueous phase contains 10% trehalose, the oil phase is corn oil - monoglyceride (9:1), and the outer aqueous phase contains 1.5% WPI - KGM complex.

[0019] As a preferred embodiment of a preparation method of an American ginseng probiotic drink of the present invention, wherein: the emulsification parameters are: high - pressure homogenization pressure 80 MPa, circulating 3 times, and solidification temperature 4°C.

[0020] As a preferred embodiment of a preparation method of an American ginseng probiotic drink of the present invention, wherein: sterilization is carried out by ultra - high - pressure treatment (600 MPa, 5 min).

[0021] As a preferred embodiment of the American ginseng probiotic drink of the present invention, wherein: the application of an American ginseng probiotic drink in the preparation of an anti - fatigue functional food.

[0022] As a preferred embodiment of the American ginseng probiotic drink of the present invention, wherein: the application of an American ginseng probiotic drink in regulating intestinal flora imbalance.

[0023] Advantages of the present invention: The present invention achieves four major breakthroughs through technological collaborative innovation:

[0024] ① In terms of ingredient retention, gradient extraction (subcritical water + ultrasound) enables the yields of saponins and polysaccharides to reach 1.5% and 8.5% respectively, with a retention rate of over 90% after 6 months, a 40% increase compared to the traditional process;

[0025] ② In terms of cell protection, SG-SA@CS microcapsules, through pore size control (<50nm) and pH-responsive swelling, increase the gastric acid survival rate from 18.5% to 87.3%, and the viable cell count during the shelf life is stable at 10 8 CFU / mL or above;

[0026] ③ In terms of functional synergy, American ginseng polysaccharides promote the proliferation of probiotics (the β-galactosidase activity is increased by 2.1 times), and the microbial metabolites (butyric acid) reversely enhance the absorption rate of saponins by 30%, achieving two-way synergy of "ingredients - microbiota";

[0027] ④ In terms of application extensibility, ultra-high pressure sterilization and WPI-KGM composite wall material technology support the diversified production of liquid beverages, solid granules and functional fortified dosage forms (such as containing ZnO NPs), and can withstand accelerated storage at 60°C (the loss of viable bacteria <15%), meeting the needs of different consumption scenarios. Detailed implementation manners

[0028] To make the above objects, features and advantages of the present invention more obvious and understandable, the following detailed description of the specific implementation manners of the present invention will be given in conjunction with the embodiments of the specification.

[0029] In the following description, many specific details are set forth to fully understand the present invention. However, the present invention can also be implemented in other ways different from those described herein. Those skilled in the art can make similar extensions without departing from the connotation of the present invention. Therefore, the present invention is not limited by the specific embodiments disclosed below.

[0030] Secondly, the so-called "one embodiment" or "embodiment" herein refers to a specific feature, structure or characteristic that can be included in at least one implementation manner of the present invention. The phrase "in one embodiment" appearing in different places in this specification does not necessarily refer to the same embodiment, nor is it an individual or selectively exclusive embodiment with other embodiments.

[0031] Example 1

[0032] This example provides an American ginseng probiotic drink and its preparation method.

[0033] Liquid American ginseng probiotic drink

[0034] Raw materials: 10% American ginseng extract (saponin content 1.5%), 4% freeze-dried powder of Lactobacillus plantarum GP258, 2% Lactobacillus salivarius LS45, 1% Bifidobacterium lactis BB12, 15% inulin, 10% whey protein, 25% SG-SA@CS microcapsule wall material, 1% N-octenyl succinic anhydride, and the balance of pure water.

[0035] Process:

[0036] American ginseng extraction: Subcritical water extraction of saponins at 120 °C → Ultrasonic extraction of polysaccharides at 50 °C → Mixing and concentrating to a solid content of 40%.

[0037] Microcapsule preparation: Inject probiotics and trehalose solution (inner aqueous phase) into SG-SA mixture (outer aqueous phase), and coaxial dripping into 2% CaCl2 solidifying solution for chitosan coating.

[0038] Mixing and filling: All components are aseptically filled after ultra-high pressure sterilization.

[0039] Viable bacteria survival rate: 87.3% in simulated gastric juice (pH 2.0, 2 h), and the viable bacteria count > 1×10^8 CFU / mL after 6 months of storage at room temperature.

[0040] Functional evaluation:

[0041] The swimming time of mice is extended by 35% (vs control group), and the number of intestinal Bifidobacterium is increased by 3.2 times.

[0042] Furthermore, the synergistic mechanism of gradient extraction technology:

[0043] Subcritical water extraction of saponins: At 120 °C and 15 MPa, the polarity of water decreases (dielectric constant drops from 78 to 40), making it easier to penetrate the cell wall of American ginseng and dissolve saponins (LogP value 1.8 - 2.5). Compared with traditional ethanol extraction, the extraction time of subcritical water is shortened by 50%, and the isomerization of Rg1 caused by high temperature is avoided.

[0044] Ultrasound-assisted extraction of polysaccharides: 50 kHz ultrasonic waves generate cavitation effect, increasing the cell wall rupture rate by 35% (observed by SEM), and accelerating the dissolution rate of polysaccharides. Compared with conventional hot water extraction, the polysaccharide yield is increased from 6.2% to 8.5% (determined by phenol-sulfuric acid method).

[0045] Gastric acid protection mechanism of SG-SA@CS microcapsules:

[0046] Structural characteristics: Sericin glue (SG) and sodium alginate (SA) are combined through Ca 2+Crosslinking forms a three-dimensional network structure (pore size <50 nm, measured by the BET method), and the chitosan (CS) coating further reduces the porosity. In an environment with pH 2.0, the swelling coefficient of the SG-SA layer is only 1.2 (vs 3.8 for the single SA layer), effectively blocking the penetration of H + penetration.

[0047] Probiotic metabolism regulation: American ginseng polysaccharide (molecular weight 20 - 50 kDa) serves as a prebiotic, promoting Lactobacillus plantarum GP258 to secrete β-galactosidase (activity increased by 2.1 times, detected by ELISA), accelerating lactose metabolism to generate short-chain fatty acids (SCFAs), reducing the pH in the capsule to 5.8 - 6.2, and forming a self-protective environment for the bacteria.

[0048] Performance verification

[0049] Control Example 1: Liquid beverage without embedded probiotics (other components are the same)

[0050] Index Example 1 Comparative Example 1 Gastric juice survival rate (2h) 87.3％ 18.5％ Viable bacteria count after storage at room temperature (6 months) <![CDATA[1.2×10 8 CFU / mL]]> <![CDATA[<1×10 6 CFU / mL]]> Saponin retention rate 92.4％ 67.8％

[0051] Using the Caco-2 cell model, the absorption rate of ginsenoside Rg1 in the Example 1 group increased by 30% compared to the monomer solution (the Papp value increased from 1.2×10 -6 cm / s to 1.56×10 -6 cm / s), proving that probiotic metabolites (such as butyric acid) can up-regulate the expression of intestinal tight junction protein ZO-1 (the expression level increased by 2.3 times as shown by Western blot).

[0052] Example 2

[0053] This example provides an American ginseng probiotic drink and its preparation method.

[0054] Solid granule type (WPI-KGM system)

[0055] Raw materials: 12% American ginseng extract, 6% compound probiotic freeze-dried powder (GP258:LS45:BB12 = 5:2:1), 20% inulin, 30% WPI-KGM microcapsule wall material, 32% maltodextrin.

[0056] Process:

[0057] Double emulsion embedding: inner aqueous phase (bacteria + 10% trehalose) → oil phase (corn oil + monoglyceride) → outer aqueous phase (1.5% WPI-KGM) → high-pressure homogenization → spray drying.

[0058] Mixing and granulation: All dry powders are granulated by fluidized bed (inlet air temperature 60°C).

[0059] Effect:

[0060] The encapsulation efficiency is 98.2%, and the viable bacteria release rate after reconstitution is >90%.

[0061] Thermal stability: During the 30-day accelerated test at 60°C, the loss of viable bacteria is <15%.

[0062] Principle analysis

[0063] Release control mechanism of WPI-KGM composite wall material:

[0064] Thermal stability: Whey protein isolate (WPI) undergoes thermal denaturation above 60°C and forms a composite gel with konjac glucomannan (KGM) through hydrophobic interaction and hydrogen bonding (the denaturation temperature detected by DSC increases from 72°C to 85°C). During spray drying, the WPI-KGM film forms a dense layer on the particle surface (the surface roughness Ra < 15 nm shown by AFM), blocking oxygen penetration (the OTR value decreases to 12 cc / m 2 ·day).

[0065] Enteric release characteristics: In simulated intestinal fluid (pH 6.8), the acetyl groups of KGM are hydrolyzed to expose free hydroxyl groups, which combine with bile salts to form micelles, promoting the swelling of the wall material (the swelling degree reaches 380%), achieving targeted release.

[0066] Optimization of double emulsion encapsulation process:

[0067] High-pressure homogenization parameters: Under a pressure of 80 MPa, the droplet size of the oil phase (corn oil / glycerol monostearate) decreases from the initial 50 μm to 1.2 μm (measured by a laser particle size analyzer), the specific surface area increases by 40 times, and the encapsulation efficiency of probiotics is improved.

[0068] Protective effect of trehalose: Adding 10% trehalose to the internal aqueous phase forms a glassy matrix during spray drying (the Tg value increases from -32°C to 45°C measured by DSC), reducing protein denaturation (the degradation rate of bacterial proteins shown by SDS-PAGE < 5%).

[0069] Performance verification

[0070] Comparative example 2: Solid granules with a single wall material (only WPI)

[0071]

[0072]

[0073] Through a DSS-induced colitis mouse model, the level of intestinal inflammatory factor IL-6 in the Example 2 group decreased by 64% (detected by ELISA), while the abundance of Bifidobacterium (measured by qPCR) increased by 2.8 times compared with Comparative example 2, proving that the WPI-KGM system can enhance the colonization effect of probiotics.

[0074] Example 3

[0075] This embodiment provides a American ginseng probiotic drink and its preparation method.

[0076] Functionally enhanced (containing ZnO nanoparticles)

[0077] Innovation point: Adding ZnO nanoparticles (10 nm, -60 mV) synthesized by probiotics to enhance antibacterial and immunomodulatory functions 3.

[0078] Process:

[0079] Preparation of ZnO NPs: The fermentation broth of Lactobacillus plantarum GP258 reacts with ZnSO4 → centrifugation and purification.

[0080] Compound: NPs (0.1%) and microencapsulated probiotics are added synchronously.

[0081] Effect:

[0082] The antibacterial zone against Staphylococcus aureus reaches 18 mm, and the inhibition rate of HT-29 cancer cells increases by 40% 3.

[0083] Immune index: The level of mouse serum IgG is increased by 2.3 times.

[0084] It should be noted that: The antibacterial synergistic effect of biosynthesized ZnO NPs:

[0085] Size effect: Organic acids (such as lactic acid) produced by the metabolism of Lactobacillus plantarum GP258 reduce Zn 2+ to ZnO NPs (10 nm, observed by HR-TEM), and its high specific surface area (89 m 2 / g) enhances the generation of reactive oxygen species (ROS) (the fluorescence intensity of DCFH-DA increases by 4 times).

[0086] Membrane damage mechanism: The positive charge on the surface of ZnO NPs (Zeta potential +60 mV) binds to the bacterial cell membrane (negatively charged), triggering lipid peroxidation (the content of MDA increases by 3.5 times), and synergistically killing pathogenic bacteria with probiotic metabolites (such as antibacterial peptides).

[0087] Immunomodulatory pathway:

[0088] Activation of dendritic cells: ZnO NPs promote the maturation of DC cells through the TLR4 / NF-κB pathway (the proportion of CD86+ cells increases from 15% to 42%), and at the same time, ginsenoside Rg1 upregulates the secretion of IFN-γ (the number of spots detected by ELISPOT increases by 2.5 times), forming a Th1 / Th2 balanced immune response.

[0089] Performance verification

[0090] Comparative example 3: A similar drink without adding ZnO NPs

[0091] Index Example 3 Comparative Example 3 Inhibitory zone of Staphylococcus aureus 18mm 6mm Apoptosis rate of HT-29 cells 58.3％ 22.1％ Serum IgG level (in mice) 4.2mg / mL 1.8mg / mL

[0092] Transcriptome analysis showed that the expression of Nrf2 pathway genes (HO-1, NQO1) in the intestinal epithelial cells of mice in Example 3 group was up-regulated by 3-5 folds, confirming that ZnO NPs and American ginseng components synergistically activated the antioxidant defense system.

[0093] In summary, through the "gradient extraction - microcapsule embedding - ultra-high pressure sterilization" trinity process, the retention rate of active ingredients (saponins > 90%), the survival rate of probiotics (in gastric acid environment > 85%) and the shelf-life stability (viable count > 10 8 CFU / mL) were synchronously optimized, which was improved compared with the traditional scheme.

[0094] Bidirectional action mechanism of "American ginseng polysaccharide - probiotic - metabolite":

[0095] Positive promotion: American ginseng polysaccharide (20 - 50 kDa) as a prebiotic shortened the logarithmic growth phase of Lactobacillus plantarum by 2 hours (β-galactosidase activity ↑ 2.1 folds);

[0096] Reverse synergistic effect: The metabolite of the flora (butyric acid) up-regulated the expression of intestinal ZO-1 protein (↑ 2.3 folds) and promoted the transmembrane absorption of saponin Rg1 (Papp value ↑ 30%).

[0097] Industrial dimension: Breaking the limitation of single dosage form, through the compatibility design of WPI-KGM composite wall material and ultra-high pressure sterilization process, the flexible production of liquid beverages, solid granules, and functional enhanced dosage forms (such as containing ZnO NPs) was realized to meet the multi-scenario needs of e-commerce, medical special diets, etc.

[0098] 2. Verification of technical advantages driven by core data

[0099] Through multi-dimensional comparison with traditional schemes and market competitors, the leading nature of the present invention was highlighted:

[0100]

[0101] The pore size of SG-SA@CS composite wall material < 50 nm (determined by BET), which was 60% lower than that of single sodium alginate (> 100 nm), and the H + permeation rate was slowed down by 4 times (fluorescent probe method);

[0102] Quantification of synergistic effect: In the Caco-2 cell model, the absorption rate of saponin Rg1 in the present invention group was increased by 30% compared with the monomer solution, and the butyric acid concentration (determined by GC-MS) was positively correlated with the expression level of ZO-1 protein (R 2 = 0.93);

[0103] Industrial feasibility: The viable cell loss rate of ultra-high pressure sterilization (600 MPa, 5 min) is reduced to less than 15% compared to pasteurization (75 °C, 15 s), and the equipment is compatible with the existing beverage production line (transformation cost < 20%).

[0104] By replacing the core components (such as replacing American ginseng with Ganoderma lucidum, Astragalus membranaceus, etc.), develop sub-products for diseases such as insomnia and metabolic syndrome;

[0105] Utilize the pH / enzyme dual-responsive characteristics of microcapsules as an alternative to antibiotics for Helicobacter pylori inhibition;

[0106] Combined with intestinal flora detection technology, customize the ratio of probiotics (such as increasing the proportion of Bifidobacterium lactis for enhancing children's immunity).

[0107] 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 preferred embodiments, those of ordinary skill in the art should understand that the technical solutions of the present invention can be modified or equivalently replaced without departing from the spirit and scope of the technical solutions of the present invention, and they should all be covered within the scope of the claims of the present invention.

Claims

1. A American ginseng probiotic drink, characterized in that: It contains the following components in weight percentage: 5-15% of American ginseng extract, 3-8% of compound probiotic freeze-dried powder, 10-25% of inulin, 5-15% of whey protein, 10-30% of microcapsule wall material, 0.5-2% of N-octenyl succinic anhydride, and the balance is purified water or maltodextrin.

2. The American ginseng probiotic drink according to claim 1, wherein: The compound probiotic contains Lactobacillus plantarum GP258, Lactobacillus salivarius LS45, and Bifidobacterium lactis BB12, and the mass ratio is (4-6):(2-3):(1-2).

3. The American ginseng probiotic drink according to claim 2, wherein: The microcapsule wall material is selected from any one of the following: a) Mulberry silk gum-sodium alginate-chitosan complex (SG:SA:CS = 4:6:2); b) Whey protein-konjac glucomannan complex (WPI:KGM = 3:1).

4. The American ginseng probiotic drink according to claim 1, characterized in that: The preparation method of the American ginseng extract includes a gradient separation step of extracting saponins by subcritical water and extracting polysaccharides by ultrasonic-assisted extraction.

5. The preparation method of an American ginseng probiotic drink according to claim 1, characterized in that: It includes the following steps: (1) Preparation of American ginseng extract; (2) Probiotic microencapsulation; (3) Component mixing and sterilization; (4) Filling or drying and forming.

6. The preparation method of an American ginseng probiotic drink according to claim 5, characterized in that: Step (2) adopts the W / O / W double emulsion method, the inner aqueous phase contains 10% trehalose, the oil phase is corn oil-monoglyceride (9:1), and the outer aqueous phase contains 1.5% WPI-KGM complex.

7. The preparation method of a kind of American ginseng probiotic drink as described in claim 5, wherein: The emulsification parameters are: high-pressure homogenization pressure 80 MPa, circulating 3 times, and curing temperature 4 °C.

8. The preparation method of an American ginseng probiotic drink according to claim 5, characterized in that: Sterilization is carried out by ultra-high pressure treatment (600 MPa, 5 min).

9. The application of the American ginseng probiotic drink as described in claim 1 in the preparation of anti-fatigue functional foods.

10. The application of the American ginseng probiotic drink as described in claim 1 in regulating intestinal flora imbalance.

Images