Fermentation type haematococcus compound health-care beverage for diminishing inflammation and resisting bacteria and preparation technology of fermentation type haematococcus compound health-care beverage

Abstract

The invention provides a fermented haematococcus compound health-care beverage for diminishing inflammation and resisting bacteria and a preparation process thereof, and belongs to the technical field of preparation of health-care beverages. The preparation process comprises the following steps: preparing the fermented haematococcus and echinacea extract; preparing a microcapsule; and preparing the composite health-care beverage. According to the invention, the fermented haematococcus solution and the Echinacea purpurea extract extracted under the assistance of high-voltage pulsed electric field treatment are compounded to prepare microcapsules, and then the microcapsules, the sucrose, the oligosaccharide, the cyclodextrin, the sweetening agent and the freshly squeezed fruit juice are granulated to prepare the granule composite health-care drink which has good anti-inflammatory and antibacterial effects.

Description

Technical Field

[0001] The invention relates to the technical field of health drink preparation, and in particular to a fermented Haematococcus algae composite health drink for anti-inflammatory and antibacterial purposes and a preparation process thereof. Background Art

[0002] In recent years, with the improvement of people's health awareness and changes in lifestyle, the incidence of inflammatory diseases and bacterial infections has been on the rise. Traditional anti-inflammatory and antibacterial drugs such as antibiotics have many problems, such as increased drug resistance caused by abuse and significant side effects, which makes people pay more attention to natural, safe and effective anti-inflammatory and antibacterial health products.

[0003] As a convenient and easy-to-absorb health care method, health drinks are favored by more and more consumers. There are many health drinks with different functions on the market, but there are relatively few functional drinks in terms of anti-inflammatory and antibacterial effects, and the efficacy and safety of some products need to be improved, so there is a large market gap and development potential.

[0004] Haematococcus pluvialis is a microalgae rich in a variety of bioactive substances, such as astaxanthin, carotenoids, polysaccharides, etc. These ingredients have multiple physiological functions such as antioxidant, anti-inflammatory, antibacterial and immunomodulatory. In particular, astaxanthin, whose antioxidant capacity is 550 times that of vitamin E, can effectively scavenge free radicals and protect cells from oxidative damage. However, there are some products based on Haematococcus pluvialis on the market, but most of them are limited to simple extraction or concentration, and fail to fully utilize the full potential of Haematococcus pluvialis, limiting the widespread application of Haematococcus pluvialis in health drinks. In addition, health drinks that only use Haematococcus pluvialis extracts, although they have certain anti-inflammatory and antibacterial effects, are limited in terms of anti-inflammatory and antibacterial effects, and it is difficult to meet people's needs for comprehensive health care.

[0005] Therefore, we proposed a fermented Haematococcus algae composite health drink for anti-inflammatory and antibacterial purposes and its preparation process. The drink not only retains the original nutritional components of Haematococcus algae, but also enhances its anti-inflammatory and antibacterial effects through the fermentation process, and further improves the anti-inflammatory and antibacterial effects through the compounding of ingredients. Summary of the invention

[0006] In view of the deficiencies in the prior art, the object of the present invention is to provide a fermented Haematococcus algae composite health drink for anti-inflammatory and antibacterial purposes and a preparation process thereof.

[0007] A preparation process of a fermented Haematococcus algae composite health drink for anti-inflammatory and antibacterial purposes comprises the following steps: S1: Preparation of fermented Haematococcus and Echinacea extracts Lactobacillus brevis and Kluyveromyces marxianus are used to synergistically ferment Haematococcus algae to prepare a fermented Haematococcus algae solution, fresh Echinacea purpurea leaves and flower buds in the bud stage are dried and crushed, added into deionized water, and then subjected to high-voltage pulse electric field treatment, and then subjected to alcohol extraction to obtain Echinacea purpurea extract; S2: Preparation of microcapsules The fermented Haematococcus solution and the Echinacea purpurea extract were embedded in the solution with gum arabic and chitosan, and cross-linked with tannic acid to prepare microcapsules with a particle size of 40-45 μm. S3: Preparation of compound health drinks The granule composite health drink is obtained by granulating 5-8 parts by weight of sucrose, 1-2 parts by weight of oligosaccharides, 2-3 parts by weight of cyclodextrin, 0.002-0.003 parts by weight of sweetener, 3-4 parts by weight of microcapsules and 5-8 parts by weight of freshly squeezed fruit juice as raw materials.

[0008] Furthermore, step S1 of preparing the fermented Haematococcus and Echinacea extracts specifically comprises the following steps: S1.1: Inoculate Lactobacillus brevis in a liquid culture medium at an inoculum rate of 3-4% (v / v), and culture at 35°C for 24 hours for activation, and activate twice to obtain an activated solution, and inoculate the activated solution in a liquid culture medium at an inoculum rate of 3-4% (v / v), and culture at 37°C for 24 hours to obtain a Lactobacillus brevis bacterial solution; S1.2: inoculating Kluyveromyces marxianus in a liquid culture medium at an inoculation rate of 2-3% (v / v), culturing at 37°C for 36 hours for activation, and continuously activating twice to obtain an activation solution, inoculating the activation solution in a liquid culture medium at an inoculation rate of 2-3% (v / v), culturing at 37°C for 36 hours, and obtaining a Kluyveromyces marxianus bacterial solution; S1.3: Prepare fermentation medium: 15 g / L glucose, 5 g / L glycerol, 30 g / L soy peptone, 3 g / L yeast extract, 1 g / L potassium dihydrogen phosphate, trace elements, vitamins and 1 L water; S1.4: 20-30 parts by weight of Haematococcus pluvialis are subjected to wall-breaking and crushing treatment to obtain Haematococcus pluvialis powder, the Haematococcus pluvialis powder is added to a fermentation medium, and then 6-8% (v / v) of a mixed bacterial solution is inoculated, wherein the mixed bacterial solution is composed of a Kluyveromyces marxianus bacterial solution and a Lactobacillus brevis bacterial solution in a volume ratio of 2-3:1-2, and the inoculation is followed by fermentation to obtain a Haematococcus pluvialis fermentation solution; S1.5: centrifuging the Haematococcus algae fermentation liquid at 4000-5000 r / min for 15-20 min to obtain a supernatant, and filtering the supernatant to obtain a fermented Haematococcus algae solution; S1.6: Collect fresh Echinacea leaves and buds in the bud stage, wash them with running water, dry them at 50-60° C. for 10-12 hours, and then grind them to obtain Echinacea powder; S1.7: adding 20-30 parts by weight of Echinacea powder to 70-80 parts by weight of deionized water, and then treating with a high voltage pulse electric field at 20-30 kV / cm and 4-6 pulses for 1-2 hours to obtain a pulse-treated Echinacea solution; S1.8: Add ethanol to the pulse-treated Echinacea solution to make its concentration reach 20-30%, then adjust the pH to 11-12 with calcium hydroxide, soak for 1-2 hours, and then reflux extraction 2-3 times, each time for 2-3 hours. After filtering, mix the filtrate to obtain the Echinacea extract.

[0009] Furthermore, the vitamins in step S1.3 consist of vitamin B1: 0.5 mg / L and vitamin B12: 0.05 mg / L.

[0010] Furthermore, the trace elements in step S1.3 consist of 0.05 g / L ferrous sulfate, 0.01 g / L manganese sulfate and 0.01 g / L zinc sulfate.

[0011] Furthermore, the fermentation conditions in step S1.4 are: dissolved oxygen 35-38%, stirring speed 200-300 rpm, pH 7.5, temperature 37-38° C., and time 72-75 h.

[0012] Furthermore, the preparation of microcapsules in step S2 specifically includes the following steps: S2.1: preparing a 5-6% concentration of gum arabic solution and a 5-6% concentration of chitosan solution, adding 10-20 parts by weight of fermented Haematococcus solution and 5-8 parts by weight of Echinacea extract to 50-60 parts by weight of the gum arabic solution, and emulsifying at 10000-12000 rpm for 50-60 seconds to obtain an emulsion; S2.2: 10-12 parts by weight of chitosan solution was added dropwise to the emulsion at a speed of 4-5 mL / min, and the mixture was stirred at 400-500 rpm for 15-20 min, and then 100-200 parts by weight of deionized water was added. The mixture was stirred at 500-600 rpm for 15-20 min, and the pH was adjusted to 4.5. The mixture was stirred at 400-500 rpm for 15-20 min to obtain a mixed system. S2.3: The mixed system is cooled to 8-10°C by ice bath, and then 3-5 parts by weight of 5% tannic acid aqueous solution is added. After stirring at 600-700 rpm for 60-70 min, the temperature is slowly raised to 45-50°C, and stirring is continued at 600-700 rpm for 60-70 min. The reaction is completed to obtain microcapsules with a particle size of 40-45 μm.

[0013] Furthermore, step S3 of preparing the composite health drink specifically comprises the following steps: S3.1: After disinfection and sterilization, 5-8 parts by weight of sucrose, 1-2 parts by weight of oligosaccharides, 2-3 parts by weight of cyclodextrin and 0.002-0.003 parts by weight of sweetener are ground and sieved through a 40-50 mesh sieve, and then dried at 60-70° C. for 20-30 min to obtain the raw materials; S3.2: After mixing all the raw materials, add 3-4 parts by weight of microcapsules and 5-8 parts by weight of freshly squeezed fruit juice, mix evenly, granulate, and then dry to obtain a granular composite health drink.

[0014] Furthermore, the freshly squeezed fruit juice in step S3.2 is one of strawberry juice, apple juice, pineapple juice and grape juice.

[0015] A fermented Haematococcus algae composite health drink for anti-inflammatory and antibacterial purposes, which is prepared by any of the above-mentioned preparation processes for a fermented Haematococcus algae composite health drink for anti-inflammatory and antibacterial purposes.

[0016] Compared with the prior art, the present invention has at least the following beneficial effects: 1. The present invention uses Kluyveromyces marxianus and Lactobacillus brevis to synergistically ferment Haematococcus. Kluyveromyces marxianus and Lactobacillus brevis have different metabolic pathways and products during the fermentation process. Kluyveromyces marxianus can ferment lactose and galactose to produce ethanol and carbon dioxide, and can also produce some flavor compounds and bioactive substances. Lactobacillus brevis mainly produces lactic acid through lactic acid fermentation, lowers the pH value, and creates an environment that is not conducive to the growth of harmful microorganisms. The synergistic fermentation of these two bacteria can form a complex microecological environment, in which the metabolic products of yeast can provide nutrition for lactic acid bacteria. , and the lactic acid produced by lactic acid bacteria can promote the growth of yeast. This interaction not only improves the fermentation efficiency, but also enhances the stability and functionality of the fermentation products. Synergistic fermentation can significantly improve the anti-inflammatory and antibacterial effects of Haematococcus pluvialis. The lactic acid and other organic acids produced during the fermentation process can lower the pH value and inhibit the growth of harmful microorganisms. At the same time, the biologically active substances produced by Kluyveromyces cerevisiae, such as polyphenols and carotenoids, have antioxidant and anti-inflammatory effects, and Lactobacillus brevis can also produce antibacterial substances such as bacteriocins during its growth process. The combined action of both parties can enhance the anti-inflammatory and antibacterial health functions of Haematococcus pluvialis.

[0017] 2. The present invention uses Echinacea extract and fermented Haematococcus algae solution in combination, which can reduce inflammation from different angles and targets. Echinacea extract focuses on regulating immune cells and cytokines, while fermented Haematococcus algae solution mainly exerts its effects through anti-oxidation and inhibition of inflammation-related enzymes. This synergistic effect is like intervening from both the "inside" (immunomodulation) and "outside" (antioxidation and enzyme inhibition) of the inflammatory response, which can more effectively reduce the symptoms of inflammation and achieve better anti-inflammatory effects at lower doses, reducing the side effects that may be caused by the high-dose use of a single ingredient. When Echinacea extract is used in combination with fermented Haematococcus algae solution, it can simultaneously inhibit the growth of multiple bacteria, including common pathogens such as Staphylococcus aureus and Escherichia coli. Moreover, their synergistic effect may also reduce the risk of bacteria developing resistance to antibacterial ingredients, thereby enabling the health drink to achieve effective anti-inflammatory and antibacterial effects.

[0018] 3. The present invention uses a high-voltage pulse electric field to treat Echinacea powder, and then performs ethanol extraction. After the Echinacea powder is treated with a high-voltage pulse electric field, the cell membrane permeability of the Echinacea plant cells can be increased. Under the action of a high-voltage pulse electric field of 20-30 kV / cm and 4-6 pulses, this "barrier" is temporarily destroyed, and the effective ingredients in the cells, such as alkylamides, polysaccharides and caffeic acid derivatives, are more likely to flow out into the surrounding solution. Since the pulse treatment has released some ingredients into the solution, ethanol can extract these ingredients more efficiently, thereby improving the overall extraction efficiency. The pressure pulse electric field treatment is relatively mild. Compared with traditional high-temperature, long-time boiling and other extraction methods, it has less damage to the activity of the effective ingredients in the cells during the process of breaking the cell wall, and can well maintain the natural active structure of these ingredients, so that the subsequently extracted ingredients can better play a role in health functions such as anti-inflammatory and antibacterial. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] The accompanying drawings, which are incorporated herein and constitute a part of the specification, illustrate embodiments of the invention and, together with the description, further serve to explain the principles of the invention and to enable those skilled in the relevant art to make and use the invention.

[0020] Figure 1 The present invention is a flow chart of the preparation process of a fermented Haematococcus algae composite health drink for anti-inflammatory and antibacterial purposes. DETAILED DESCRIPTION

[0021] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0022] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0023] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the invention pertains. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention.

[0024] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0025] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0026] Example 1: A preparation process of a fermented Haematococcus algae composite health drink for anti-inflammatory and antibacterial purposes, such as Figure 1 As shown, the following steps are included: S1: Preparation of fermented Haematococcus and Echinacea extracts S1.1: Inoculate Lactobacillus brevis in a liquid culture medium at an inoculum rate of 3% (v / v), culture at 35°C for 24 hours for activation, activate twice in succession to obtain an activated solution, inoculate the activated solution in a liquid culture medium at an inoculum rate of 3% (v / v), culture at 37°C for 24 hours, and obtain a Lactobacillus brevis bacterial solution; S1.2: inoculating Kluyveromyces marxianus in a liquid culture medium at an inoculum volume of 2% (v / v), culturing at 37°C for 36 hours for activation, and performing activation twice to obtain an activation solution, inoculating the activation solution in a liquid culture medium at an inoculum volume of 2% (v / v), culturing at 37°C for 36 hours, and obtaining a Kluyveromyces marxianus bacterial solution; S1.3: Prepare fermentation medium: 15 g / L glucose, 5 g / L glycerol, 30 g / L soy peptone, 3 g / L yeast extract, 1 g / L potassium dihydrogen phosphate, trace elements, vitamins and 1 L water; Trace elements are ferrous sulfate 0.05 g / L, manganese sulfate 0.01 g / L and zinc sulfate 0.01 g / L; Vitamins: vitamin B1: 0.5 mg / L and vitamin B12: 0.05 mg / L; S1.4: 20 parts by weight of Haematococcus pluvialis are subjected to a wall-breaking and pulverizing treatment to obtain Haematococcus pluvialis powder, the Haematococcus pluvialis powder is added to a fermentation medium, and then a 6% (v / v) mixed bacterial solution is inoculated, wherein the mixed bacterial solution is composed of a Kluyveromyces marxianus bacterial solution and a Lactobacillus brevis bacterial solution in a volume ratio of 2:1, and the inoculation is followed by fermentation to obtain a Haematococcus pluvialis fermentation solution; The fermentation conditions were: dissolved oxygen 35%, stirring speed 200 rpm, pH 7.5, temperature 37°C, and time 72 h; S1.5: centrifuging the Haematococcus algae fermentation liquid at 4000 r / min for 15 min to obtain a supernatant, and filtering the supernatant to obtain a fermented Haematococcus algae solution; S1.6: Collect fresh Echinacea leaves and buds in the bud stage, wash them with running water, dry them at 50° C. for 10 h, and then grind them to obtain Echinacea powder; S1.7: 20 parts by weight of Echinacea powder are added to 70 parts by weight of deionized water, and then treated in a high-voltage pulse electric field at 20 kV / cm and 4 pulses for 1 hour to obtain a pulse-treated Echinacea solution; S1.8: adding ethanol to the pulse-treated Echinacea solution to a concentration of 20%, adjusting the pH to 11 with calcium hydroxide, soaking for 1 hour, and then performing reflux extraction twice, each time for 2 hours, filtering, and mixing the filtrate to obtain an Echinacea extract; S2: Preparation of microcapsules S2.1: preparing a 5% concentration of gum arabic solution and a 5% concentration of chitosan solution, adding 10 parts by weight of fermented Haematococcus solution and 5 parts by weight of Echinacea extract to 50 parts by weight of the gum arabic solution, and emulsifying at 10000 rpm for 50 seconds to obtain an emulsion; S2.2: 10 parts by weight of chitosan solution was added dropwise to the emulsion at a rate of 4 mL / min, and stirred at 400 rpm for 15 min, and then 100 parts by weight of deionized water was added. The mixture was stirred at 500 rpm for 15 min, and the pH was adjusted to 4.5. The mixture was stirred at 400 rpm for 15 min to obtain a mixed system. S2.3: The mixed system was cooled to 8°C by ice bath, and then 3 parts by weight of 5% tannic acid aqueous solution was added, and the mixture was stirred at 600 rpm for 60 min, and then the temperature was slowly raised to 45°C, and the mixture was stirred at 600 rpm for 60 min. The reaction was terminated to obtain microcapsules with a particle size of 45 μm. S3: Preparation of compound health drinks S3.1: 5 parts by weight of sucrose, 1 part by weight of isomalto-oligosaccharide, 2 parts by weight of cyclodextrin and 0.002 parts by weight of glucose sweetener are disinfected and sterilized, then ground and pulverized through a 40-mesh sieve, and then dried at 60° C. for 20 min to obtain the raw materials; S3.2: After mixing all the raw materials, add 3 parts by weight of microcapsules and 5 parts by weight of freshly squeezed pineapple juice, mix evenly, granulate, and then dry to obtain a granular composite health drink.

[0027] Example 2: A preparation process of a fermented Haematococcus algae composite health drink for anti-inflammatory and antibacterial purposes, such as Figure 1 As shown, the following steps are included: S1: Preparation of fermented Haematococcus and Echinacea extracts S1.1: Inoculate Lactobacillus brevis in a liquid culture medium at an inoculum rate of 4% (v / v), and culture at 35°C for 24 hours for activation, and activate twice to obtain an activated solution, and inoculate the activated solution in a liquid culture medium at an inoculum rate of 4% (v / v), and culture at 37°C for 24 hours to obtain a Lactobacillus brevis bacterial solution; S1.2: inoculating Kluyveromyces marxianus in a liquid culture medium at an inoculum volume of 3% (v / v), culturing at 37°C for 36 hours for activation, and performing activation twice to obtain an activation solution, inoculating the activation solution in a liquid culture medium at an inoculum volume of 3% (v / v), culturing at 37°C for 36 hours, and obtaining a Kluyveromyces marxianus bacterial solution; S1.3: Prepare fermentation medium: 15 g / L glucose, 5 g / L glycerol, 30 g / L soy peptone, 3 g / L yeast extract, 1 g / L potassium dihydrogen phosphate, trace elements, vitamins and 1 L water; Trace elements are ferrous sulfate 0.05 g / L, manganese sulfate 0.01 g / L and zinc sulfate 0.01 g / L; Vitamins: vitamin B1: 0.5 mg / L and vitamin B12: 0.05 mg / L; S1.4: 30 parts by weight of Haematococcus pluvialis are subjected to a wall-breaking and pulverizing treatment to obtain Haematococcus pluvialis powder, the Haematococcus pluvialis powder is added to a fermentation medium, and then 8% (v / v) of a mixed bacterial solution is inoculated, wherein the mixed bacterial solution is composed of a Kluyveromyces marxianus bacterial solution and a Lactobacillus brevis bacterial solution in a volume ratio of 3:2, and the inoculation is followed by fermentation to obtain a Haematococcus pluvialis fermentation solution; The fermentation conditions were: dissolved oxygen 38%, stirring speed 200 rpm, pH 7.5, temperature 37°C, and time 72 h; S1.5: centrifuging the Haematococcus algae fermentation liquid at 4000 r / min for 15 min to obtain a supernatant, and filtering the supernatant to obtain a fermented Haematococcus algae solution; S1.6: Collect fresh Echinacea leaves and buds in the bud stage, wash them with running water, dry them at 50° C. for 10 h, and then grind them to obtain Echinacea powder; S1.7: 30 parts by weight of Echinacea powder is added to 80 parts by weight of deionized water, and then treated in a high-voltage pulse electric field of 20 kV / cm and 4 pulses for 1 hour to obtain a pulse-treated Echinacea solution; S1.8: adding ethanol to the pulse-treated Echinacea solution to a concentration of 30%, adjusting the pH to 12 with calcium hydroxide, soaking for 1 hour, and then performing reflux extraction twice, each time for 2 hours, filtering, and mixing the filtrate to obtain an Echinacea extract; S2: Preparation of microcapsules S2.1: prepare a 6% concentration of gum arabic solution and a 6% concentration of chitosan solution, add 20 parts by weight of fermented Haematococcus solution and 8 parts by weight of Echinacea extract to 60 parts by weight of the gum arabic solution, and emulsify at 10000 rpm for 50 seconds to obtain an emulsion; S2.2: 12 parts by weight of chitosan solution was added dropwise to the emulsion at a rate of 4 mL / min, and the mixture was stirred at 400 rpm for 15 min, and then 200 parts by weight of deionized water was added. The mixture was stirred at 500 rpm for 15 min, and the pH was adjusted to 4.5. The mixture was stirred at 400 rpm for 15 min to obtain a mixed system. S2.3: The mixed system was cooled to 8°C by ice bath, and then 5 parts by weight of 5% tannic acid aqueous solution was added, and the mixture was stirred at 600 rpm for 60 min, and then the temperature was slowly raised to 45°C, and the mixture was stirred at 600 rpm for 60 min. The reaction was terminated to obtain microcapsules with a particle size of 40 μm. S3: Preparation of compound health drinks S3.1: 8 parts by weight of sucrose, 2 parts by weight of isomalto-oligosaccharide, 3 parts by weight of cyclodextrin and 0.003 parts by weight of glucose sweetener are disinfected and sterilized, then ground and crushed through a 40-mesh sieve, and then dried at 60° C. for 20 min to obtain the raw materials; S3.2: After mixing all the raw materials, add 4 parts by weight of microcapsules and 8 parts by weight of freshly squeezed strawberry juice, mix evenly, granulate, and then dry to obtain a granular composite health drink.

[0028] Example 3: A preparation process of a fermented Haematococcus algae composite health drink for anti-inflammatory and antibacterial purposes, such as Figure 1 As shown, the following steps are included: S1: Preparation of fermented Haematococcus and Echinacea extracts S1.1: Inoculate Lactobacillus brevis in a liquid culture medium at an inoculum rate of 3% (v / v), culture at 35°C for 24 hours for activation, activate twice in succession to obtain an activated solution, inoculate the activated solution in a liquid culture medium at an inoculum rate of 3% (v / v), culture at 37°C for 24 hours, and obtain a Lactobacillus brevis bacterial solution; S1.2: inoculating Kluyveromyces marxianus in a liquid culture medium at an inoculum volume of 2% (v / v), culturing at 37°C for 36 hours for activation, and performing activation twice to obtain an activation solution, inoculating the activation solution in a liquid culture medium at an inoculum volume of 2% (v / v), culturing at 37°C for 36 hours, and obtaining a Kluyveromyces marxianus bacterial solution; S1.3: Prepare fermentation medium: 15 g / L glucose, 5 g / L glycerol, 30 g / L soy peptone, 3 g / L yeast extract, 1 g / L potassium dihydrogen phosphate, trace elements, vitamins and 1 L water; Trace elements are ferrous sulfate 0.05 g / L, manganese sulfate 0.01 g / L and zinc sulfate 0.01 g / L; Vitamins: vitamin B1: 0.5 mg / L and vitamin B12: 0.05 mg / L; S1.4: 20 parts by weight of Haematococcus pluvialis are subjected to a wall-breaking and pulverizing treatment to obtain Haematococcus pluvialis powder, the Haematococcus pluvialis powder is added to a fermentation medium, and then a 6% (v / v) mixed bacterial solution is inoculated, wherein the mixed bacterial solution is composed of a Kluyveromyces marxianus bacterial solution and a Lactobacillus brevis bacterial solution in a volume ratio of 2:1, and the inoculation is followed by fermentation to obtain a Haematococcus pluvialis fermentation solution; The fermentation conditions were: dissolved oxygen 35%, stirring speed 300 rpm, pH 7.5, temperature 38°C, and time 75 h; S1.5: centrifuging the Haematococcus algae fermentation liquid at 5000 r / min for 20 min to obtain a supernatant, and filtering the supernatant to obtain a fermented Haematococcus algae solution; S1.6: Collect fresh Echinacea leaves and buds in the bud stage, wash them with running water, dry them at 60° C. for 12 h, and then grind them to obtain Echinacea powder; S1.7: 20 parts by weight of Echinacea powder are added to 70 parts by weight of deionized water, and then treated in a high-voltage pulse electric field at 30 kV / cm and 6 pulses for 2 hours to obtain a pulse-treated Echinacea solution; S1.8: adding ethanol to the pulse-treated Echinacea solution to a concentration of 20%, adjusting the pH to 11 with calcium hydroxide, soaking for 2 hours, and then performing reflux extraction 3 times, each time for 3 hours, filtering, and mixing the filtrate to obtain an Echinacea extract; S2: Preparation of microcapsules S2.1: preparing a 5% concentration of gum arabic solution and a 5% concentration of chitosan solution, adding 20 parts by weight of fermented Haematococcus solution and 5 parts by weight of Echinacea extract to 50 parts by weight of the gum arabic solution, and emulsifying at 12000 rpm for 60 seconds to obtain an emulsion; S2.2: 10 parts by weight of chitosan solution was added dropwise to the emulsion at a rate of 5 mL / min, and stirred at 500 rpm for 20 min. Then, 100 parts by weight of deionized water was added. The mixture was stirred at 600 rpm for 20 min, and the pH was adjusted to 4.5. The mixture was stirred at 500 rpm for 20 min to obtain a mixed system. S2.3: The mixed system was cooled to 10°C by ice bath, and then 3 parts by weight of 5% tannic acid aqueous solution was added, and the mixture was stirred at 700 rpm for 70 min, and then the temperature was slowly raised to 50°C, and the mixture was stirred at 700 rpm for 70 min. The reaction was terminated to obtain microcapsules with a particle size of 43 μm. S3: Preparation of compound health drinks S3.1: 5 parts by weight of sucrose, 1 part by weight of isomalto-oligosaccharide, 2 parts by weight of cyclodextrin and 0.002 parts by weight of glucose sweetener are disinfected and sterilized, then ground and pulverized through a 50-mesh sieve, and then dried at 70° C. for 30 min to obtain the raw materials; S3.2: After mixing all the raw materials, add 3 parts by weight of microcapsules and 5 parts by weight of freshly squeezed apple juice, mix evenly, granulate, and then dry to obtain a granular composite health drink.

[0029] Comparative Example 1: Compared with Example 1, the difference of Comparative Example 1 is that Comparative Example 1 removes the Lactobacillus brevis liquid in steps S1.1 and S1.4, and the other steps remain unchanged to prepare the granular composite health drink, which is recorded as Comparative Example 1.

[0030] Comparative Example 2: Compared with Example 1, the difference of Comparative Example 2 is that the Kluyveromyces marxianus bacterial solution in steps S1.2 and S1.4 is removed, and the other steps remain unchanged to prepare the granular composite health drink, which is recorded as Comparative Example 2.

[0031] Comparative Example 3: Compared with Example 1, the difference of Comparative Example 3 is that Comparative Example 3 removes the Echinacea extract in steps S1.5-S1.8 and S2.1, and the other steps remain unchanged to prepare the granular compound health drink, which is recorded as Comparative Example 3.

[0032] Comparative Example 4: Compared with Example 1, the difference of Comparative Example 4 is that Comparative Example 4 removes the fermented Haematococcus solution in steps S1.1-S1.5 and S3.1, and the other steps remain unchanged to prepare the granular composite health drink, which is recorded as Comparative Example 4.

[0033] Comparative Example 5: Compared with Example 1, the difference of Comparative Example 5 is that Comparative Example 5 removes the high-voltage pulse electric field treatment in step S1.7, and replaces the Echinacea solution after pulse treatment in step S1.8 with Echinacea solution, and the other steps remain unchanged to prepare a granular compound health drink, which is recorded as Comparative Example 5.

[0034] 10 g of the granular composite health drink prepared in Examples 1-3 and Comparative Examples 1-5 was added to 50 mL of 37° C. warm water to obtain a composite health drink.

[0035] Preparation of simulated gastric fluid: 8.3 g / L peptone, 3.5 g / L anhydrous glucose, 2.05 g / L sodium chloride, 0.6 g / L potassium dihydrogen phosphate, 0.11 g / L calcium chloride, 0.37 g / L potassium chloride, 10 g / L pepsin. Sterilize the gastric juice at high pressure at 121°C for 30 min, adjust the pH to 3.5 with dilute hydrochloric acid to obtain simulated gastric juice.

[0036] 10 mL of the composite health drinks prepared in Examples 1-3 and Comparative Examples 1-5 were added to 100 mL of simulated gastric fluid. After 10 minutes of simulated digestion, the digestive fluids were aspirated as test samples.

[0037] Inhibition of TNF-α expression: Fresh blood was treated with EDTA as anticoagulant, blood cells were separated by Ficoll, and the cells were resuspended in RIMP1640 medium containing 10% fetal bovine serum. 100 mL of 1×10 5 cells / mL Freshly isolated cells, total number of cells per well: 10 4 Three wells were made for each sample.

[0038] Take the test sample, add sterile PBS solution to prepare the test sample stock solution (10 mg / mL), and then use sterile PBS to dilute the stock solution to the corresponding working concentration for use; add 10 mL of the test solution of the corresponding working concentration into the wells of the culture plate.

[0039] The culture plate was placed in an incubator at 37°C containing 5% CO2 for 15 minutes; 10 mL of 100 mg / mL LPS was added and the plate was incubated in an incubator at 37°C containing 5% CO2 for 16 hours.

[0040] The culture plate was centrifuged at 1000 rpm for 15 min, the supernatant was transferred to a new culture plate, and the TNF-α concentration in the culture supernatant was determined using an ELISA reader; or the culture plate was frozen at 20°C to avoid repeated freezing and thawing, and the inhibition rate was calculated.

[0041] Antibacterial effect: Preparation of bacterial plates: Take a flat double dish with a diameter of about 90mm and a height of 16mm, and inject 20ml of 60℃ sterile No. Ⅲ culture medium (prepared according to the first part of the 2010 edition of the Chinese Veterinary Pharmacopoeia) into each dish, spread it evenly on the bottom of the dish, and place it on a water table to solidify as the bottom layer. Take another appropriate amount of culture medium, heat and melt it, cool it to 48℃, add 5ml of Escherichia coli suspension, and spread it evenly on the bottom layer as the bacterial layer. Place it on a water table to cool, and then make a bacterial plate.

[0042] The above digestive fluids were made into drug-sensitive paper sheets and the sizes of the inhibition zones were determined.

[0043] Method for determining the size of the inhibition zone: Take sterilized filter paper pieces with a diameter of 0.6 cm and soak them in different digestive solutions for 40 minutes. Take them out for use. Place drug-sensitive paper pieces on the prepared bacterial plates with aseptic operation, 6 pieces for each drug. Then place the plates in a 37°C incubator and culture them for 24 hours. After that, measure the size of the inhibition zone and take the average value.

[0044] The specific test results are shown in Table 1 below: Table 1. Inhibition of TNF-α expression and inhibition zone test results Inhibition rate of TNF-α release / % Inhibition zone (mm) Example 1 94.3 5.84 Example 2 93.1 5.75 Example 3 93.7 5.79 Comparative Example 1 84.7 4.34 Comparative Example 2 85.3 4.52 Comparative Example 3 75.2 3.23 Comparative Example 4 67.3 3.74 Comparative Example 5 86.3 4.93 It can be seen from the data in Table 1 that the composite health drink prepared by the present invention has excellent anti-inflammatory effect. It can be seen from the data of Comparative Examples 1 and 2 that the use of Kluyveromyces marxianus and Lactobacillus brevis for synergistic fermentation of Haematococcus can enhance the anti-inflammatory and antibacterial effect of Haematococcus, thereby increasing the anti-inflammatory and antibacterial effect of the composite health drink. It can be seen from the results of Comparative Examples 3 and 4 that the combined use of Echinacea extract and fermented Haematococcus solution can more effectively inhibit the expression of TNF-α, thereby alleviating inflammatory symptoms, and significantly inhibiting the growth of bacteria, so that the health drink can achieve effective anti-inflammatory and antibacterial effects; it can be seen from the data of Comparative Example 5 that under the action of the high-voltage pulsed electric field, the effective ingredients in the cells are more easily flowed into the surrounding solution, and the activity of the effective ingredients in the cells is less damaged, and the natural active structure of these ingredients can be well maintained, so that the ingredients extracted subsequently can better play a role in health functions such as anti-inflammatory and antibacterial.

[0045] The above embodiments are merely illustrative of the principles and effects of the present invention, and are not intended to limit the present invention. Anyone familiar with the art may modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by a person of ordinary skill in the art without departing from the spirit and technical concept disclosed by the present invention shall still be covered by the claims of the present invention.

Claims

1. A preparation process of a fermented Haematococcus algae composite health drink for anti-inflammatory and antibacterial purposes, characterized in that: The steps include: S1: Preparation of fermented Haematococcus and Echinacea extracts Lactobacillus brevis and Kluyveromyces marxianus are used to synergistically ferment Haematococcus algae to prepare a fermented Haematococcus algae solution, fresh Echinacea purpurea leaves and flower buds in the bud stage are dried and crushed, added into deionized water, and then subjected to high-voltage pulse electric field treatment, and then subjected to alcohol extraction to obtain Echinacea purpurea extract; S2: Preparation of microcapsules The fermented Haematococcus solution and the Echinacea purpurea extract were embedded in the solution with gum arabic and chitosan, and cross-linked with tannic acid to prepare microcapsules with a particle size of 40-45 μm. S3: Preparation of compound health drinks The granule composite health drink is obtained by granulating 5-8 parts by weight of sucrose, 1-2 parts by weight of oligosaccharides, 2-3 parts by weight of cyclodextrin, 0.002-0.003 parts by weight of sweetener, 3-4 parts by weight of microcapsules and 5-8 parts by weight of freshly squeezed fruit juice as raw materials.

2. The preparation process of a fermented Haematococcus algae composite health drink for anti-inflammatory and antibacterial purposes according to claim 1, characterized in that: Step S1: Preparation of fermented Haematococcus and Echinacea extracts, specifically comprising the following steps: S1.1: Inoculate Lactobacillus brevis in a liquid culture medium at an inoculum rate of 3-4% (v / v), and culture at 35°C for 24 hours for activation, and activate twice to obtain an activated solution, and inoculate the activated solution in a liquid culture medium at an inoculum rate of 3-4% (v / v), and culture at 37°C for 24 hours to obtain a Lactobacillus brevis bacterial solution; S1.2: inoculating Kluyveromyces marxianus in a liquid culture medium at an inoculation rate of 2-3% (v / v), culturing at 37°C for 36 hours for activation, and continuously activating twice to obtain an activation solution, inoculating the activation solution in a liquid culture medium at an inoculation rate of 2-3% (v / v), culturing at 37°C for 36 hours, and obtaining a Kluyveromyces marxianus bacterial solution; S1.3: Prepare fermentation medium: 15 g / L glucose, 5 g / L glycerol, 30 g / L soy peptone, 3 g / L yeast extract, 1 g / L potassium dihydrogen phosphate, trace elements, vitamins and 1 L water; S1.4: 20-30 parts by weight of Haematococcus pluvialis are subjected to wall-breaking and crushing treatment to obtain Haematococcus pluvialis powder, the Haematococcus pluvialis powder is added to a fermentation medium, and then 6-8% (v / v) of a mixed bacterial solution is inoculated, wherein the mixed bacterial solution is composed of a Kluyveromyces marxianus bacterial solution and a Lactobacillus brevis bacterial solution in a volume ratio of 2-3:1-2, and the inoculation is followed by fermentation to obtain a Haematococcus pluvialis fermentation solution; S1.5: centrifuging the Haematococcus algae fermentation liquid at 4000-5000 r / min for 15-20 min to obtain a supernatant, and filtering the supernatant to obtain a fermented Haematococcus algae solution; S1.6: Collect fresh Echinacea leaves and buds in the bud stage, wash them with running water, dry them at 50-60° C. for 10-12 hours, and then grind them to obtain Echinacea powder; S1.7: adding 20-30 parts by weight of Echinacea powder to 70-80 parts by weight of deionized water, and then treating with a high voltage pulse electric field at 20-30 kV / cm and 4-6 pulses for 1-2 hours to obtain a pulse-treated Echinacea solution; S1.8: Add ethanol to the pulse-treated Echinacea solution to make its concentration reach 20-30%, then adjust the pH to 11-12 with calcium hydroxide, soak for 1-2 hours, and then reflux extraction 2-3 times, each time for 2-3 hours. After filtering, mix the filtrate to obtain the Echinacea extract.

3. The preparation process of a fermented Haematococcus algae composite health drink for anti-inflammatory and antibacterial purposes according to claim 2, characterized in that: The vitamins in step S1.3 consist of vitamin B1: 0.5 mg / L and vitamin B12: 0.05 mg / L.

4. The preparation process of a fermented Haematococcus algae composite health drink for anti-inflammatory and antibacterial purposes according to claim 2, characterized in that: The trace elements in step S1.3 consist of 0.05 g / L ferrous sulfate, 0.01 g / L manganese sulfate and 0.01 g / L zinc sulfate.

5. The preparation process of a fermented Haematococcus algae composite health drink for anti-inflammatory and antibacterial purposes according to claim 2, characterized in that: The fermentation conditions in step S1.4 are: dissolved oxygen 35-38%, stirring speed 200-300 rpm, pH 7.5, temperature 37-38° C., and time 72-75 h.

6. The preparation process of a fermented Haematococcus algae composite health drink for anti-inflammatory and antibacterial purposes according to claim 5, characterized in that: Step S2: Preparation of microcapsules, specifically comprising the following steps: S2.1: preparing a 5-6% concentration of gum arabic solution and a 5-6% concentration of chitosan solution, adding 10-20 parts by weight of fermented Haematococcus solution and 5-8 parts by weight of Echinacea extract to 50-60 parts by weight of the gum arabic solution, and emulsifying at 10000-12000 rpm for 50-60 seconds to obtain an emulsion; S2.2: 10-12 parts by weight of chitosan solution was added dropwise to the emulsion at a speed of 4-5 mL / min, and the mixture was stirred at 400-500 rpm for 15-20 min, and then 100-200 parts by weight of deionized water was added. The mixture was stirred at 500-600 rpm for 15-20 min, and the pH was adjusted to 4.

5. The mixture was stirred at 400-500 rpm for 15-20 min to obtain a mixed system. S2.3: The mixed system is cooled to 8-10°C by ice bath, and then 3-5 parts by weight of 5% tannic acid aqueous solution is added. After stirring at 600-700 rpm for 60-70 min, the temperature is slowly raised to 45-50°C, and stirring is continued at 600-700 rpm for 60-70 min. The reaction is completed to obtain microcapsules with a particle size of 40-45 μm.

7. The preparation process of a fermented Haematococcus algae composite health drink for anti-inflammatory and antibacterial purposes according to claim 6, characterized in that: Step S3: Preparation of the composite health drink, specifically comprising the following steps: S3.1: After disinfection and sterilization, 5-8 parts by weight of sucrose, 1-2 parts by weight of oligosaccharides, 2-3 parts by weight of cyclodextrin and 0.002-0.003 parts by weight of sweetener are ground and sieved through a 40-50 mesh sieve, and then dried at 60-70° C. for 20-30 min to obtain the raw materials; S3.2: After mixing all the raw materials, add 3-4 parts by weight of microcapsules and 5-8 parts by weight of freshly squeezed fruit juice, mix evenly, granulate, and then dry to obtain a granular composite health drink.

8. The preparation process of the fermented Haematococcus algae composite health drink for anti-inflammatory and antibacterial purposes according to claim 7, characterized in that: The freshly squeezed fruit juice in step S3.2 is one of strawberry juice, apple juice, pineapple juice and grape juice.

9. A fermented Haematococcus algae composite health drink for anti-inflammatory and antibacterial purposes, characterized in that: The beverage is prepared by the preparation process of a fermented Haematococcus algae composite health drink for anti-inflammatory and antibacterial purposes as described in any one of claims 1 to 8.

Images