A malus hallings fermenting liquid promoting telomerase activity

Abstract

The present application relates to the technical field of biological fermentation, and discloses a roxburgh rose fermentation liquor for promoting telomerase activity, which comprises the following raw materials in terms of weight components: 200-240 parts of roxburgh rose juice, and 24-150 parts of aqueous apigenin solution; wherein the 200-240 parts of roxburgh rose juice in terms of weight components is prepared by mixing 85-180 parts of roxburgh rose fruit and 60-115 parts of distilled water in terms of weight components, and then pressing the mixture into juice by using a juicer; the 24-150 parts of aqueous apigenin solution in terms of weight components is prepared by mixing 10-25 parts of apigenin and 14-125 parts of active water in terms of weight components, and then stirring the mixture into solution; the roxburgh rose juice and the aqueous apigenin solution are intermittently and alternately mixed, and 10-45 parts of compound microbial bodies in terms of weight components are added; and the mixture is placed in a fermentation chamber to perform fermentation, so as to obtain the roxburgh rose fermentation liquor. The roxburgh rose fermentation liquor simultaneously increases the content of vitamin C, polyphenols and flavonoids, and increases the activity of superoxide dismutase (SOD), and reduces the content of fructose, and thus exhibits the effect of promoting telomerase activity.

Description

Technical Field

[0001] This invention relates to the field of bio-fermentation technology, and specifically to a prickly pear fermentation broth that promotes telomerase activity. Background Technology

[0002] Prickly pear fermented liquid is an essential liquid obtained by pressing prickly pear fruit, which is unique to Guizhou Province. Prickly pear fruit can be used as medicine and has the effects of strengthening the stomach, aiding digestion, nourishing, and stopping diarrhea. In particular, prickly pear is rich in superoxide dismutase (SOD), which has anti-aging, anti-cancer, antiviral, and anti-radiation effects. It is widely used in the prevention and treatment of cardiovascular, digestive system, and various tumor diseases.

[0003] Currently, among the publicly available research on the preparation and application of prickly pear fermentation broth in China, for example, Chinese patent document (publication number CN109105889A) discloses a preparation process and application of fermented prickly pear juice, which utilizes a compound lactic acid bacteria including Lactobacillus plantarum, Lactobacillus paracasei, Lactobacillus casei, and Acetobacter to ferment prickly pear juice, thereby significantly enhancing the DPPH free radical scavenging ability of the prickly pear fermentation broth and the immunity of mice.

[0004] Meanwhile, a method for fermenting golden prickly pear is disclosed in Chinese patent document (publication number CN108523109B), which uses a mixture of strains, including Lactobacillus casei strain, Bifidobacterium longum strain and Saccharomyces cerevisiae strain, to ferment golden prickly pear samples and obtain a golden prickly pear fermentation broth with enhanced antioxidant capacity.

[0005] However, current prickly pear fermentation methods only consider the fermentation capacity of the strains. The fermentation broth obtained from existing bacterial fermentation only contains substances such as vitamin C, superoxide dismutase (SOD), and polyphenols that are carried by the strains themselves. It is impossible to simultaneously increase the content of vitamin C, polyphenols, and flavonoids, as well as increase the activity of superoxide dismutase (SOD), reduce the fructose content, and show a promoting effect on telomerase activity. Summary of the Invention

[0006] The present invention aims to provide a prickly pear fermentation broth that promotes telomerase activity, in order to solve the problem that the existing prickly pear fermentation broth obtained by bacterial fermentation only contains substances such as vitamin C, superoxide dismutase (SOD), and polyphenols that are carried by the bacteria themselves, and cannot simultaneously increase the content of vitamin C, polyphenols and flavonoids, as well as increase the activity of superoxide dismutase (SOD) and reduce the fructose content, thus exhibiting the effect of promoting telomerase activity.

[0007] To achieve the above objectives, the present invention adopts the following technical solution: 1) A prickly pear fermentation broth that promotes telomerase activity, comprising the following raw materials by weight: 200-240 parts prickly pear juice by weight and 24-150 parts apigenin aqueous solution by weight; wherein, the prickly pear juice by weight of 200-240 parts prickly pear fruit by weight of 85-180 parts prickly pear fruit and distilled water by weight of 60-115 parts by weight are mixed and pressed into juice by a juicer, and the apigenin aqueous solution by weight of 24-150 parts apigenin by weight of 10-25 parts apigenin and active water by weight of 14-125 parts by weight are mixed and stirred into a solution, the prickly pear juice and apigenin aqueous solution are intermittently mixed and 10-45 parts by weight of compound bacteria are added, and fermentation is carried out in a fermentation chamber to obtain prickly pear fermentation broth.

[0008] This technical solution involves mixing distilled water and prickly pear fruit and pressing the mixture into juice using a juicer. Since distilled water is free of electrolytes and free ions, the process of mixing distilled water and prickly pear fruit for juicing greatly reduces impurities in the prickly pear juice and the generation of harmful substances through chemical reactions. This significantly increases the content of prickly pear nutrients in the juice.

[0009] Based on this, in order to further increase the content of vitamin C, polyphenols and flavonoids in the prickly pear fermentation broth, as well as to increase the activity of superoxide dismutase (SOD), an aqueous solution of apigenin was intermittently mixed with prickly pear juice. Apigenin (4',5,7-trihydroxyflavone) is a dietary flavonoid. During the mixing of the apigenin aqueous solution and the prickly pear juice, the flavonoid content in the prickly pear fermentation broth was increased simultaneously. In addition, apigenin regulates the signaling cascade of cyclic adenosine monophosphate (cAMP) response element-binding protein (CREB), brain-derived neurotrophic factor (BDNF), cAMP-CREB-BDNF and n-methyl-d-aspartate (NMDA) receptors. Therefore, the prickly pear fermentation broth fermented with the mixture of apigenin aqueous solution and prickly pear juice promotes telomerase activity. Telomerase activity is an enzyme that can extend the length of human chromosome telomeres, thereby repairing cells and prolonging cell life.

[0010] A apigenin solution is prepared by mixing 10-25 parts by weight of apigenin and 14-125 parts by weight of active water. Active water itself has the characteristic of promoting the absorption of nutrients. Therefore, the apigenin solution containing active water can promote the absorption of nutrients in prickly pear juice during fermentation, effectively and simultaneously increasing the content of vitamin C, polyphenols and flavonoids, as well as increasing the activity of superoxide dismutase (SOD).

[0011] In addition, because the juice of prickly pear contains a variety of flavonoids, the fermentation process of the juice produces flavonoid aglycones. These flavonoid aglycones combine with dihydromyricetin, which is found in apigenin, to enhance the properties of the flavonoid aglycones. Together, they possess a variety of biological functions, including free radical scavenging, anti-oxidation, antihypertensive, antithrombotic, antitumor, and anti-inflammatory effects.

[0012] 2) A prickly pear fermentation broth that promotes telomerase activity according to 1), wherein: The method for preparing the prickly pear juice includes the following steps: Step S1-1, Rinsing: Rinse the harvested prickly pears to remove dust and other impurities from the surface of the prickly pears after harvesting, so that the surface of the prickly pears is free of impurities. Step S1-2, Screening: The prickly pears rinsed in step S1-1 are sent to a screening and washing machine to remove the thorns, sepals and leaves from the prickly pears and obtain the prickly pear fruits. Step S1-3, Juicing: The prickly pear fruit washed in step S1-2 is fed into a juicer, and distilled water of matching weight is added to the juicer. After the prickly pear fruit and distilled water are mixed, the juicer is started to crush and juice the fruit. After crushing and juicing, the juice is filtered through a filter membrane to obtain prickly pear juice.

[0013] In this technical solution, the prickly pear juice is produced through a production process of rinsing, screening, and juicing. Distilled water is free of electrolytes and free ions. Therefore, when distilled water and prickly pear fruit are mixed and juiced, impurities in the prickly pear juice and the generation of harmful substances through chemical reactions between substances are greatly reduced. This results in prickly pear juice with high nutritional content, full release of nutrients, high utilization rate of prickly pear fruit, unique aroma, refreshing taste, and rich in various vitamins, amino acids, and superoxide dismutase (SOD).

[0014] 3) A prickly pear fermentation broth that promotes telomerase activity as described in 1), wherein: The method for preparing the apigenin aqueous solution includes the following steps: Step S2-1: Take dried celery seeds and put them into a grinder to grind them into powder. Add a certain amount of organic solvent to the celery seed powder to remove volatile oil compounds from the celery seed powder and obtain celery seed powder. Step S2-2: Add an organic solvent to the celery seed powder from step S2-1. The organic solvent effectively removes the volatile oil compounds in the celery seed powder to obtain preliminary celery seed powder particles. Step S2-3: At room temperature, the celery seed powder particles from step S2-2 are soaked in an ethanol-water solution by circulating reflux to obtain celery seed liquid. The celery seed liquid is then concentrated under reduced pressure to obtain apigenin.

[0015] In this technical solution, when celery seeds are crushed in step S2-1, the celery seeds are crushed in a dry state. The water content of the dried celery seeds is greatly reduced, so that the celery seed powder formed after crushing is in a dry state. This makes it easier to mix the dried celery seed powder with organic solvents, and the volatile oil compounds in the celery seed powder can be effectively removed, thus preserving the apigenin in the celery seed powder.

[0016] At room temperature, celery seed powder particles from step S2-2 were soaked in an ethanol-water solution via a circulating reflux method to obtain celery seed liquid. The celery seed liquid was then concentrated under reduced pressure, and the purified apigenin obtained after extraction had a purity of 98%. Apigenin has effects such as regulating blood pressure and blood lipids, anti-oxidation, improving immunity, and delaying aging. It alleviates Alzheimer's disease by reducing neuroinflammation, protecting neurons from oxidative stress, and enhancing neuronal survival and synaptic plasticity. Furthermore, purchased celery seeds were used as the main raw material, with a purchase price of 1.5 yuan / jin. Therefore, the low purchase price of celery seeds resulted in a low overall cost for apigenin preparation. Simultaneously, the repeated extraction of apigenin using an ethanol-water solution via a circulating reflux method reduced waste of celery seed powder particles while increasing the apigenin extraction rate.

[0017] 4) A prickly pear fermentation broth that promotes telomerase activity as described in 3), wherein the celery seed powder particles from step S2-2 are soaked in an ethanol-water solution by a circulating reflux soaking method to obtain celery seed liquid, and the circulating soaking method is a flow-type circulating soaking in a soaking tank: The soaking tank includes a tank body filled with an ethanol-water solution. A spirally coiled soaking pipe is provided along the height of the tank body. Several staggered flow holes are evenly distributed along the length of the soaking pipe. The diameter of the celery seed powder particles is larger than the diameter of the flow holes. One end of the soaking pipe passes through the tank body and connects to a feeding pipe. The top of the feeding pipe has an inlet for adding celery seed powder particles. The other end of the soaking pipe extends towards the side of the tank body and connects to a circulation pipe. The other end of the circulation pipe extends to the side wall of the feeding pipe and connects to the feeding pipe. A suction pump is installed on the circulation pipe.

[0018] Before use, the tank designed in this technical solution is filled with an ethanol-water solution through the opening at the top of the tank. The opening is then sealed with a cap hinged to the tank. After the tank is filled with ethanol-water, celery seed powder particles are added to the inlet of the feeding pipe. The celery seed powder particles enter the soaking pipe along the feeding pipe. Because the soaking pipe has several flow holes evenly distributed along its length, the ethanol-water solution enters the soaking pipe through these flow holes. After the celery seed powder particles enter the soaking pipe through the feeding pipe, because the diameter of the celery seed powder particles is larger than the diameter of the flow holes, the celery seed powder particles cannot flow into the tank through the flow holes. They can only mix with the ethanol-water solution in the soaking pipe and soak until all the celery seed powder particles have completely entered the soaking pipe. Then, the tank is left to stand for fifteen minutes.

[0019] After soaking all the celery seed powder particles in the ethanol-water solution for 15 minutes, start the suction pump on the circulation pipe. The negative pressure generated by the suction pump on the circulation pipe will draw the ethanol-water solution containing the celery seed powder particles from the soaking pipe. Since the circulation pipe is located on the outside of the tank, the ethanol-water solution containing the celery seed powder particles enters the feeding pipe through the circulation pipe, and then enters the soaking tank again through the feeding pipe. During this process, the suction pump is started for 10 minutes, allowing the celery seed powder particles to circulate in the circulation pipe, feeding pipe, and soaking pipe for 10 minutes, so that the celery seed powder particles are fully soaked and mixed with the ethanol-water solution. Then, let the solution containing the celery seed powder particles in the soaking pipe stand for 15 minutes, and then restart the suction pump. Repeat the above process three to four times.

[0020] After repeating the above process three to four times, the celery seed powder particles gradually dissolve in the ethanol-water solution to obtain celery seed liquid. The valve on the discharge pipe located on the side of the tank is then opened. Since the discharge pipe is connected to the bottom side of the tank, the celery seed liquid is discharged from the tank after the valve is opened. The celery seed liquid is then concentrated under reduced pressure to obtain apigenin. Celery seed liquid is obtained through a circulating reflux soaking method, and the purity of the refined apigenin after extraction is 98%.

[0021] 5) A prickly pear fermentation broth that promotes telomerase activity as described in 1), wherein prickly pear juice and apigenin aqueous solution are intermittently and alternately mixed and 10-45 parts by weight of compound microbial cells are added, and fermentation is carried out in a fermentation chamber to obtain the prickly pear fermentation broth, comprising the following steps: Step 3-1: The prickly pear juice and apigenin solution, accounting for 10% of their total weight components and 2% of their total weight components, are added to the fermentation tank for the first time. The prickly pear juice and apigenin solution are stirred and mixed by stirring with a stirring rod. Step 3-2: Based on the initial mixing of prickly pear juice and apigenin solution in the fermentation tank in Step 3-1, prickly pear juice at 20% of its total weight and apigenin solution at 10% of its total weight are added to the fermentation tank a second time. The prickly pear juice and apigenin solution are mixed by stirring with a stirring rod. After the second mixing, prickly pear juice at 30% of its total weight and apigenin solution at 15% of its total weight are added to the fermentation tank each time. The mixture is stirred with a stirring rod after each addition. Step 3-3: After intermittently mixing the prickly pear juice and apigenin solution in the fermentation tank, add 10-45 parts of compound bacteria by weight to the fermentation tank and stir the mixture with a stirring rod. Then place the fermentation tank in the fermentation environment to ferment and obtain prickly pear fermentation liquid.

[0022] This technical solution combines prickly pear juice and apigenin aqueous solution with a complex microbial culture for fermentation. This not only transforms the macromolecular substances and astringent substances in prickly pear and apigenin, but also increases the content of vitamin C, superoxide dismutase (SOD), and polyphenols during the fermentation process, which is beneficial for digestion and absorption. At the same time, for the first time, the prickly pear juice and apigenin aqueous solution are mixed in the fermentation tank at a ratio of 10% of the total weight of the prickly pear juice and 2% of the total weight of the apigenin aqueous solution, so that the apigenin aqueous solution dissolves in the prickly pear juice.

[0023] The prickly pear juice and apigenin solution were then stirred and mixed using a stirring rod. After the second addition and stirring, the prickly pear juice and apigenin solution were added to the fermentation tank at a ratio of 30% and 15% of their total weight, respectively. After each addition, the mixture was stirred and mixed in the fermentation tank with a stirring rod. This intermittent and alternating mixing of the prickly pear juice and apigenin solution with different amounts effectively ensured that the prickly pear juice and apigenin solution were evenly mixed. This allowed the compound bacteria and apigenin solution to effectively inhibit the oxidation of vitamin C and polyphenols in the prickly pear juice, effectively maintaining a high content of vitamin C and polyphenols in the prickly pear juice during long-term fermentation.

[0024] 6) A prickly pear fermentation broth that promotes telomerase activity according to 5), wherein: The method of using the compound bacterial culture in step 3-3 includes the following steps: Step 3-3 (1): The compound microbial body includes 2-5 parts by weight of yeast, 6-12 parts by weight of lactic acid bacteria and 3-9 parts by weight of acetic acid bacteria. Yeast and lactic acid bacteria are inoculated into a liquid mixed with apigenin aqueous solution and the first fermentation is carried out for 20-72 hours. After the fermentation is completed, the residue is removed by filtration to obtain the first fermentation liquid. Acetic acid bacteria are inoculated into the first fermentation liquid to obtain the second fermentation liquid. Step 3-3 (2): Add the prickly pear juice to the second fermentation liquid in step 3-3 (1), pulverize, let stand, filter to remove residue, and obtain slurry. Inoculate the slurry with acetic acid bacteria to obtain fermentation liquid. Continue fermentation until the mass concentration of alcohol in the fermentation liquid drops below 5‰, and then end the fermentation to obtain prickly pear fermentation liquid.

[0025] In this technical solution, the compound microbial body includes yeast, lactic acid bacteria, and acetic acid bacteria. The first fermentation involves inoculating yeast and lactic acid bacteria into an aqueous solution of apigenin, resulting in a first fermentation broth from the mixture of apigenin, yeast, and lactic acid bacteria. Simultaneously, acetic acid bacteria are inoculated into the first fermentation broth to obtain a second fermentation broth. After adding the second fermentation broth to the prickly pear juice, the yeast in the second fermentation broth rapidly ferments the prickly pear juice, effectively limiting unwanted bacteria in the fermentation broth while removing bitter components from both the prickly pear juice and the apigenin solution, thus retaining their nutritional components.

[0026] In the second fermentation broth, lactic acid bacteria can increase the content of flavonoids and polyphenols in the prickly pear juice during fermentation, while slightly reducing the content of polysaccharides. At the same time, the antioxidant activity, DPPH free radical scavenging rate, and absorbability of the prickly pear fermentation broth are improved. Combined with the acetic acid bacteria in the second fermentation broth, the fermentation process of the prickly pear juice not only does not consume the superoxide dismutase (SOD) in the prickly pear fermentation broth, but also increases the activity of superoxide dismutase (SOD) in the prickly pear fermentation broth after fermentation, greatly enhancing the activity of superoxide dismutase (SOD) in the prickly pear fermentation broth.

[0027] 7) A prickly pear fermentation broth that promotes telomerase activity according to 5), wherein: The fermentation environment is characterized by a fermentation temperature controlled at 25-37℃ and a fermentation time controlled at 120-320 days.

[0028] Our company's research focuses on optimizing fermentation process parameters. The optimal fermentation temperature is 28℃, with a range of 25-37℃. The optimal fermentation time is 283 days, with a range of 120-320 days. When the fermentation temperature is above 28℃, the compound microbial culture allows for sufficient fermentation of the prickly pear juice and apigenin solution. Fermentation ends when the alcohol concentration in the prickly pear fermentation liquid drops below 5‰. In this case, the fermentation time is less than the optimal 283 days. Conversely, if the temperature is below 28℃, the fermentation time is greater than the optimal 283 days.

[0029] When the fermentation time is controlled to be more than 283 days, the compound microbial culture can fully ferment the prickly pear juice and apigenin solution. Fermentation ends when the alcohol concentration in the prickly pear fermentation broth drops below 5‰. At this point, the fermentation temperature is lower than the optimal fermentation temperature of 28℃; conversely, the fermentation temperature is higher than the optimal fermentation temperature of 28℃. A comparison of the vitamin C, polyphenol, and flavonoid content in this prickly pear fermentation broth revealed that the prickly pear fermentation broth prepared by the above method showed a simultaneous increase in vitamin C, polyphenol, and flavonoid content, a decrease in fructose content, and an increase in superoxide dismutase (SOD) activity, demonstrating a promoting effect on telomerase activity.

[0030] 8) A prickly pear fermentation broth that promotes telomerase activity according to 1), wherein: The 200 parts by weight of prickly pear juice, 85 parts by weight of prickly pear fruit and 115 parts by weight of distilled water are mixed and pressed into juice by a juicer. The 24 parts by weight of apigenin aqueous solution, 10 parts by weight of apigenin and 14 parts by weight of active water are mixed and stirred into a solution.

[0031] 9) A prickly pear fermentation broth that promotes telomerase activity according to 1), wherein: The 225 parts by weight of prickly pear juice, 152 parts by weight of prickly pear fruit, and 73 parts by weight of distilled water were mixed and pressed into juice using a juicer. The 140 parts by weight of apigenin aqueous solution, 19 parts by weight of apigenin, and 121 parts by weight of active water were mixed and stirred into a solution.

[0032] 10) A prickly pear fermentation broth that promotes telomerase activity according to 1), wherein: The 240 parts by weight of prickly pear juice, 180 parts by weight of prickly pear fruit, and 60 parts by weight of distilled water are mixed and pressed into juice using a juicer. The 150 parts by weight of apigenin aqueous solution, 25 parts by weight of apigenin, and 125 parts by weight of active water are mixed and stirred into a solution.

[0033] Compared with the prior art, the present invention also has the following technical effects: The prickly pear fermentation liquid in this invention contains a variety of flavonoids. During the fermentation process, flavonoids will produce flavonoid aglycones. Both flavonoid aglycones and apigenin contain dihydromyricetin. Therefore, the dihydromyricetin produced during the co-fermentation of prickly pear juice and apigenin aqueous solution complement each other and have a variety of biological functions, including free radical scavenging, anti-oxidation, antihypertensive, antithrombotic, antitumor, and anti-inflammatory effects. Attached Figure Description

[0034] Figure 1 This is a flowchart illustrating the preparation process of a prickly pear fermentation broth that promotes telomerase activity according to the present invention. Figure 2 This is a flowchart illustrating the preparation of prickly pear juice from prickly pear fermentation broth that promotes telomerase activity, according to the present invention. Figure 3 This is a flowchart illustrating the preparation of an aqueous solution of apigenin from prickly pear fermentation broth that promotes telomerase activity, according to the present invention. Figure 4 This is a schematic diagram of the structure of a soaking tank for preparing an aqueous solution of apigenin from prickly pear fermentation broth that promotes telomerase activity, according to the present invention. Detailed Implementation

[0035] The following detailed description illustrates the specific implementation method: The reference numerals in the accompanying drawings include: 1. Tank body; 2. Immersion pipe; 3. Feed pipe; 4. Inlet; 5. Circulation pipe; 6. Suction pump; 7. Discharge pipe; 8. Valve; 9. Flow hole.

[0036] Example 1 A prickly pear fermentation broth that promotes telomerase activity comprises raw materials in the following weight components: 200 parts by weight of prickly pear juice and 24 parts by weight of apigenin aqueous solution.

[0037] In Example 1, the preparation of a prickly pear fermentation broth that promotes telomerase activity is as follows: 200 parts by weight of prickly pear juice, 85 parts by weight of prickly pear fruit, and 115 parts by weight of distilled water are mixed and pressed into juice using a juicer. 24 parts by weight of apigenin aqueous solution, 10 parts by weight of apigenin, and 14 parts by weight of active water are mixed and stirred to form a solution. (See [link to previous section]). Figure 1 As shown, the prickly pear juice and apigenin aqueous solution were intermittently mixed and 10 parts by weight of compound bacteria were added. The mixture was then placed in a fermentation chamber for fermentation to obtain prickly pear fermentation liquid.

[0038] Example 2 A prickly pear fermentation broth that promotes telomerase activity comprises raw materials in the following weight components: 225 parts by weight of prickly pear juice and 140 parts by weight of apigenin aqueous solution.

[0039] In Example 2, the preparation of a prickly pear fermentation broth that promotes telomerase activity is as follows: 225 parts by weight of prickly pear juice, 152 parts by weight of prickly pear fruit, and 73 parts by weight of distilled water are mixed and pressed into juice using a juicer. 140 parts by weight of apigenin aqueous solution, 19 parts by weight of apigenin, and 121 parts by weight of active water are mixed and stirred to form a solution. (See [link to example]). Figure 1 As shown, prickly pear juice and apigenin aqueous solution were intermittently mixed and 30 parts by weight of compound bacteria were added. The mixture was then placed in a fermentation chamber for fermentation to obtain prickly pear fermentation liquid.

[0040] Example 3 A prickly pear fermentation broth that promotes telomerase activity comprises raw materials in the following weight components: 240 parts by weight of prickly pear juice and 150 parts by weight of apigenin aqueous solution.

[0041] In Example 3, the preparation of a prickly pear fermentation broth that promotes telomerase activity is as follows: 240 parts by weight of prickly pear juice, 180 parts by weight of prickly pear fruit, and 60 parts by weight of distilled water are mixed and pressed into juice using a juicer. 150 parts by weight of apigenin aqueous solution, 25 parts by weight of apigenin, and 125 parts by weight of active water are mixed and stirred to form a solution. (See [link to relevant documentation]). Figure 1 As shown, the prickly pear juice and apigenin aqueous solution were intermittently mixed and 45 parts by weight of compound bacteria were added, and the mixture was placed in a fermentation chamber for fermentation to obtain prickly pear fermentation liquid.

[0042] The preparation methods of prickly pear juice in Examples 1-3, combined with Figure 2 As shown, it includes the following steps: Step S1-1, Rinsing: Rinse the harvested prickly pears to remove dust and other impurities from the surface of the prickly pears after harvesting, so that the surface of the prickly pears is free of impurities. Step S1-2, Screening: The prickly pears rinsed in step S1-1 are sent to a screening and washing machine to remove the thorns, sepals and leaves from the prickly pears and obtain the prickly pear fruits. Step S1-3, Juicing: The prickly pear fruit washed in step S1-2 is fed into a juicer, and distilled water of matching weight is added to the juicer. After the prickly pear fruit and distilled water are mixed, the juicer is started to crush and juice the fruit. After crushing and juicing, the juice is filtered through a filter membrane to obtain prickly pear juice.

[0043] In Examples 1-3, the prickly pear juice was produced through a process of rinsing, screening, and juicing. Distilled water was used as the electrolyte-free and free ion-free water. Therefore, when distilled water and prickly pear fruit were mixed and juiced, impurities in the prickly pear juice and the generation of harmful substances through chemical reactions between substances were greatly reduced. This resulted in a prickly pear juice with high nutritional content, full release of nutrients, high utilization rate of prickly pear fruit, unique aroma, refreshing taste, and rich in various vitamins, amino acids, and superoxide dismutase (SOD).

[0044] In addition, the preparation methods of apigenin aqueous solution in Examples 1-3, combined with Figure 3 As shown, it includes the following steps: Step S2-1: Take dried celery seeds and put them into a grinder to grind them into powder. Add a certain amount of organic solvent to the celery seed powder to remove volatile oil compounds from the celery seed powder and obtain celery seed powder. Step S2-2: Add an organic solvent to the celery seed powder from step S2-1. The organic solvent effectively removes the volatile oil compounds in the celery seed powder to obtain preliminary celery seed powder particles. Step S2-3: At room temperature, the celery seed powder particles from step S2-2 are soaked in an ethanol-water solution by circulating reflux to obtain celery seed liquid. The celery seed liquid is then concentrated under reduced pressure to obtain apigenin.

[0045] In Examples 1-3, when celery seeds are pulverized in step S2-1, the celery seeds are pulverized in a dry state. The water content of the celery seeds in the dry state is greatly reduced, so that the celery seed powder formed after pulverizing the celery seeds is in a dry state. When the dry celery seed powder is mixed with an organic solvent, the volatile oil compounds in the celery seed powder can be effectively removed, thus preserving the apigenin in the celery seed powder.

[0046] At room temperature, celery seed powder particles from step S2-2 were soaked in an ethanol-water solution via a circulating reflux method to obtain celery seed liquid. The celery seed liquid was then concentrated under reduced pressure, and the purified apigenin obtained after extraction had a purity of 98%. Apigenin has effects such as regulating blood pressure and blood lipids, anti-oxidation, improving immunity, and delaying aging. It alleviates Alzheimer's disease by reducing neuroinflammation, protecting neurons from oxidative stress, and enhancing neuronal survival and synaptic plasticity. Furthermore, purchased celery seeds were used as the main raw material, with a purchase price of 1.5 yuan / jin. Therefore, the low purchase price of celery seeds resulted in a low overall cost for apigenin preparation. Simultaneously, the repeated extraction of apigenin using an ethanol-water solution via a circulating reflux method reduced waste of celery seed powder particles while increasing the apigenin extraction rate.

[0047] In the preparation method of apigenin aqueous solution in Examples 1-3, celery seed powder particles from step S2-2 are soaked in an ethanol aqueous solution by a circulating reflux soaking method to obtain celery seed liquid. The circulating reflux soaking method is a flow-type circulating soaking in a soaking tank. See Figure 4 As shown, the specific structure of the soaking tank used in Embodiments 1-3 is as follows: The soaking tank includes a tank body 1, which is filled with an ethanol aqueous solution. A spiral soaking pipe 2 is provided in the tank body 1 along its height direction. Several staggered flow holes 9 are evenly distributed along the length of the soaking pipe 2. The diameter of the celery seed powder particles is larger than the diameter of the flow holes 9. One end of the soaking pipe 2 passes through the tank body 1 and is connected to the feed pipe 3. The top end of the feed pipe 3 has an inlet 4 for adding celery seed powder particles. The other end of the soaking pipe 2 extends toward the side of the tank body 1 and is connected to the circulation pipe 5. The other end of the circulation pipe 5 extends to the side wall of the feed pipe 3 and is connected to the feed pipe 3. A suction pump 6 is installed on the circulation pipe 5.

[0048] Before use, the tank 1 designed in this technical solution is filled with an ethanol-water solution through the opening at the top of the tank 1. Then, the opening of the tank 1 is sealed by the cap hinged to the tank 1. After the ethanol-water solution is easily filled into the tank 1, celery seed powder particles are added to the inlet 4 of the feeding pipe 3. The celery seed powder particles enter the soaking pipe 2 along the feeding pipe 3. Because the soaking pipe 2 has several flow holes 9 evenly distributed along its length, the ethanol-water solution enters the soaking pipe 2 through the flow holes 9. After the celery seed powder particles enter the soaking pipe 2 through the feeding pipe 3, because the diameter of the celery seed powder particles is larger than the diameter of the flow holes 9, the celery seed powder particles cannot flow into the tank 1 through the flow holes 9. They can only mix with the ethanol-water solution in the soaking pipe 2 and soak until all the celery seed powder particles have completely entered the soaking pipe 2. Then, let it stand for fifteen minutes.

[0049] After all the celery seed powder particles are soaked in the ethanol-water solution for 15 minutes, the suction pump 6 on the circulation pipe 5 is started. The negative pressure generated by the suction pump 6 on the circulation pipe 5 draws the ethanol-water solution containing the celery seed powder particles from the soaking pipe 2. Since the circulation pipe 5 is located outside the tank 1, the ethanol-water solution containing the celery seed powder particles enters the feeding pipe 3 through the circulation pipe 5, and then enters the soaking tank again through the feeding pipe 3. During this process, the suction pump 6 is started for 10 minutes, allowing the celery seed powder particles to circulate in the circulation pipe 5, the feeding pipe 3, and the soaking pipe for 10 minutes, so that the celery seed powder particles are fully soaked and mixed with the ethanol-water solution. Then, the ethanol-water solution containing the celery seed powder particles in the soaking pipe is allowed to stand for 15 minutes, and then the suction pump 6 is restarted. The above process is repeated three to four times.

[0050] After repeating the above process three to four times, the celery seed powder particles gradually dissolve in the ethanol-water solution to obtain celery seed liquid. The valve 8 on the discharge pipe 7 located on the side of tank 1 is then opened. Since the discharge pipe 7 is connected to the bottom side of tank 1, opening the valve 8 on the side of tank 1 allows the celery seed liquid inside tank 1 to be discharged. The celery seed liquid is then concentrated under reduced pressure to obtain apigenin. Celery seed liquid is obtained through a circulating reflux soaking method, and the purity of the refined apigenin after extraction is 98%.

[0051] In Examples 1-3, prickly pear juice and apigenin aqueous solution were intermittently and alternately mixed, and 10-45 parts by weight of compound microbial cells were added. The mixture was then placed in a fermentation chamber for fermentation to obtain prickly pear fermentation broth, including the following steps: Step 3-1: The prickly pear juice and apigenin solution, accounting for 10% of their total weight components and 2% of their total weight components, are added to the fermentation tank for the first time. The prickly pear juice and apigenin solution are stirred and mixed by stirring with a stirring rod. Step 3-2: Based on the initial mixing of prickly pear juice and apigenin solution in the fermentation tank in Step 3-1, prickly pear juice at 20% of its total weight and apigenin solution at 10% of its total weight are added to the fermentation tank a second time. The prickly pear juice and apigenin solution are mixed by stirring with a stirring rod. After the second mixing, prickly pear juice at 30% of its total weight and apigenin solution at 15% of its total weight are added to the fermentation tank each time. The mixture is stirred with a stirring rod after each addition. Step 3-3: After intermittently mixing the prickly pear juice and apigenin solution in the fermentation tank, add 10-45 parts of compound bacteria by weight to the fermentation tank and stir the mixture with a stirring rod. Then place the fermentation tank in the fermentation environment to ferment and obtain prickly pear fermentation liquid.

[0052] Fermenting prickly pear juice and apigenin solution with a complex microbial culture not only transforms the macromolecular substances and astringent substances in prickly pear and apigenin, but also increases the content of vitamin C, superoxide dismutase (SOD), and polyphenols during the fermentation process, which is beneficial to digestion and absorption. At the same time, for the first time, the prickly pear juice and apigenin solution are mixed in the fermentation tank at a ratio of 10% of the total weight of the prickly pear juice and 2% of the total weight of the apigenin solution, so that the apigenin solution dissolves in the prickly pear juice.

[0053] The prickly pear juice and apigenin solution were then stirred and mixed using a stirring rod. After the second addition and stirring, the prickly pear juice and apigenin solution were added to the fermentation tank at a ratio of 30% and 15% of their total weight, respectively. After each addition, the mixture was stirred and mixed in the fermentation tank with a stirring rod. This intermittent and alternating mixing of the prickly pear juice and apigenin solution with different amounts effectively ensured that the prickly pear juice and apigenin solution were evenly mixed. This allowed the compound bacteria and apigenin solution to effectively inhibit the oxidation of vitamin C and polyphenols in the prickly pear juice, effectively maintaining a high content of vitamin C and polyphenols in the prickly pear juice during long-term fermentation.

[0054] The method of using the compound bacterial culture in step 3-3 includes the following steps: Step 3-3 (1): The compound microbial body includes 2-5 parts by weight of yeast, 6-12 parts by weight of lactic acid bacteria and 3-9 parts by weight of acetic acid bacteria. Yeast and lactic acid bacteria are inoculated into a liquid mixed with apigenin aqueous solution and the first fermentation is carried out for 20-72 hours. After the fermentation is completed, the residue is removed by filtration to obtain the first fermentation liquid. Acetic acid bacteria are inoculated into the first fermentation liquid to obtain the second fermentation liquid. Step 3-3(2): Add the prickly pear juice to the second fermentation liquid in step 3-3(1), pulverize, let stand, filter to remove residue, and obtain slurry. Inoculate the slurry with acetic acid bacteria to obtain fermentation liquid. Continue fermentation until the mass concentration of alcohol in the fermentation liquid drops below 5‰, and then stop fermentation to obtain prickly pear fermentation liquid.

[0055] The compound microbial culture includes yeast, lactic acid bacteria, and acetic acid bacteria. Yeast and lactic acid bacteria are inoculated into an aqueous solution of apigenin for the first fermentation, resulting in a first fermentation broth. Acetic acid bacteria are then inoculated into the first fermentation broth to obtain a second fermentation broth. When the second fermentation broth is added to prickly pear juice, the yeast in the second fermentation broth rapidly ferments the prickly pear juice, effectively limiting unwanted bacteria in the fermentation broth and removing bitter components from both the prickly pear juice and the apigenin solution while retaining their nutritional components.

[0056] In the second fermentation broth, lactic acid bacteria can increase the content of flavonoids and polyphenols in the prickly pear juice during fermentation, while slightly reducing the content of polysaccharides. At the same time, the antioxidant activity, DPPH free radical scavenging rate, and absorbability of the prickly pear fermentation broth are improved. Combined with the acetic acid bacteria in the second fermentation broth, the fermentation of prickly pear juice not only does not consume the superoxide dismutase (SOD) in the prickly pear fermentation broth, but also increases the activity of superoxide dismutase (SOD) in the prickly pear fermentation broth after fermentation, greatly improving the activity of superoxide dismutase (SOD) in the prickly pear fermentation broth.

[0057] Furthermore, the fermentation environment is such that the fermentation temperature is controlled at 25-37℃ and the fermentation time is controlled at 120-320 days.

[0058] Our company's research focuses on optimizing fermentation process parameters. The optimal fermentation temperature is 28℃, with a range of 25-37℃. The optimal fermentation time is 283 days, with a range of 120-320 days. When the fermentation temperature is above 28℃, the compound microbial culture allows for sufficient fermentation of the prickly pear juice and apigenin solution. Fermentation ends when the alcohol concentration in the prickly pear fermentation liquid drops below 5‰. In this case, the fermentation time is less than the optimal 283 days. Conversely, if the temperature is below 28℃, the fermentation time is greater than the optimal 283 days.

[0059] When the fermentation time is controlled to be more than 283 days, the compound microbial culture can fully ferment the prickly pear juice and apigenin solution. Fermentation ends when the alcohol concentration in the prickly pear fermentation broth drops below 5‰. At this point, the fermentation temperature is lower than the optimal fermentation temperature of 28℃; conversely, the fermentation temperature is higher than the optimal fermentation temperature of 28℃. A comparison of the vitamin C, polyphenol, and flavonoid content in this prickly pear fermentation broth revealed that the prickly pear fermentation broth prepared by the above method showed a simultaneous increase in vitamin C, polyphenol, and flavonoid content, a decrease in fructose content, and an increase in superoxide dismutase (SOD) activity, demonstrating a promoting effect on telomerase activity.

[0060] The above are merely embodiments of this solution. Commonly known technical solutions and / or characteristics are not described in detail here. It should be noted that those skilled in the art can make various modifications and improvements without departing from the technical solution of this solution. These modifications and improvements should also be considered within the scope of protection of this solution, and will not affect the effectiveness of the implementation of this solution or the practicality of the patent. The scope of protection claimed in this application shall be determined by the content of its claims, and the specific embodiments described in the specification can be used to interpret the content of the claims.

Claims

1. A prickly pear fermentation broth that promotes telomerase activity, characterized in that, The raw materials include: 200-240 parts by weight of prickly pear juice and 24-150 parts by weight of apigenin aqueous solution; wherein, the 200-240 parts by weight of prickly pear juice is mixed with 85-180 parts by weight of prickly pear fruit and 60-115 parts by weight of distilled water and pressed into juice by a juicer; the 24-150 parts by weight of apigenin aqueous solution is mixed with 10-25 parts by weight of apigenin and 14-125 parts by weight of active water to form a solution; the prickly pear juice and apigenin aqueous solution are intermittently mixed and mixed with 10-45 parts by weight of compound bacteria, and fermented in a fermentation chamber to obtain prickly pear fermented liquid.

2. The prickly pear fermentation broth that promotes telomerase activity according to claim 1, characterized in that: The method for preparing the prickly pear juice includes the following steps: Step S1-1, Rinsing: Rinse the harvested prickly pears to remove dust and other impurities from the surface of the prickly pears after harvesting, so that the surface of the prickly pears is free of impurities. Step S1-2, Screening: The prickly pears rinsed in step S1-1 are sent to a screening and washing machine to remove the thorns, sepals and leaves from the prickly pears and obtain the prickly pear fruits. Step S1-3, Juicing: The prickly pear fruit washed in step S1-2 is fed into a juicer, and distilled water of matching weight is added to the juicer. After the prickly pear fruit and distilled water are mixed, the juicer is started to crush and juice the fruit. After crushing and juicing, the juice is filtered through a filter membrane to obtain prickly pear juice.

3. The prickly pear fermentation broth that promotes telomerase activity according to claim 1, characterized in that: The method for preparing the apigenin aqueous solution includes the following steps: Step S2-1: Take dried celery seeds and put them into a grinder to grind them into powder. Add a certain amount of organic solvent to the celery seed powder to remove volatile oil compounds from the celery seed powder and obtain celery seed powder. Step S2-2: Add an organic solvent to the celery seed powder from step S2-1. The organic solvent effectively removes the volatile oil compounds in the celery seed powder to obtain preliminary celery seed powder particles. Step S2-3: At room temperature, the celery seed powder particles from step S2-2 are soaked in an ethanol-water solution by circulating reflux to obtain celery seed liquid. The celery seed liquid is then concentrated under reduced pressure to obtain celery.

4. The prickly pear fermentation broth that promotes telomerase activity according to claim 3, characterized in that: In step S2-2, celery seed powder particles are soaked in an ethanol-water solution using a circulating reflux method to obtain celery seed liquid. The circulating reflux method is a flow-type circulating reflux soaking tank. The soaking tank includes a tank body filled with an ethanol-water solution. A spirally coiled soaking pipe is provided along the height of the tank body. Several staggered flow holes are evenly distributed along the length of the soaking pipe. The diameter of the celery seed powder particles is larger than the diameter of the flow holes. One end of the soaking pipe passes through the tank body and connects to a feeding pipe. The top of the feeding pipe has an inlet for adding celery seed powder particles. The other end of the soaking pipe extends towards the side of the tank body and connects to a circulation pipe. The other end of the circulation pipe extends to the side wall of the feeding pipe and connects to the feeding pipe. A suction pump is installed on the circulation pipe.

5. The prickly pear fermentation broth that promotes telomerase activity according to claim 1, characterized in that: The process involves intermittently and alternately mixing prickly pear juice and apigenin aqueous solution, adding 10-45 parts by weight of a complex microbial culture, and then fermenting the mixture in a fermentation chamber to obtain prickly pear fermentation broth. This includes the following steps: Step 3-1: The prickly pear juice and apigenin solution, accounting for 10% of their total weight components and 2% of their total weight components, are added to the fermentation tank for the first time. The prickly pear juice and apigenin solution are stirred and mixed by stirring with a stirring rod. Step 3-2: Based on the initial mixing of prickly pear juice and apigenin solution in the fermentation tank in Step 3-1, prickly pear juice at 20% of its total weight and apigenin solution at 10% of its total weight are added to the fermentation tank a second time. The prickly pear juice and apigenin solution are mixed by stirring with a stirring rod. After the second mixing, prickly pear juice at 30% of its total weight and apigenin solution at 15% of its total weight are added to the fermentation tank each time. The mixture is stirred with a stirring rod after each addition. Step 3-3: After intermittently mixing the prickly pear juice and apigenin solution in the fermentation tank, add 10-45 parts of compound bacteria by weight to the fermentation tank and stir the mixture with a stirring rod. Then place the fermentation tank in the fermentation environment to ferment and obtain prickly pear fermentation liquid.

6. The prickly pear fermentation broth that promotes telomerase activity according to claim 5, characterized in that: The method of using the compound bacterial culture in step 3-3 includes the following steps: Step 3-3 (1): The compound microbial body includes 2-5 parts by weight of yeast, 6-12 parts by weight of lactic acid bacteria and 3-9 parts by weight of acetic acid bacteria. Yeast and lactic acid bacteria are inoculated into a liquid mixed with apigenin aqueous solution and the first fermentation is carried out for 20-72 hours. After the fermentation is completed, the residue is removed by filtration to obtain the first fermentation liquid. Acetic acid bacteria are inoculated into the first fermentation liquid to obtain the second fermentation liquid. Step 3-3 (2): Add the prickly pear juice to the second fermentation liquid in step 3-3 (1), pulverize, let stand, filter to remove residue, and obtain slurry. Inoculate the slurry with acetic acid bacteria to obtain fermentation liquid. Continue fermentation until the mass concentration of alcohol in the fermentation liquid drops below 5‰, and then end the fermentation to obtain prickly pear fermentation liquid.

7. The prickly pear fermentation broth that promotes telomerase activity according to claim 5, characterized in that: The fermentation environment is characterized by a fermentation temperature controlled at 25–37°C and a fermentation time controlled at 120–320 days.

8. The prickly pear fermentation broth that promotes telomerase activity according to claim 1, characterized in that: The 200 parts by weight of prickly pear juice, 85 parts by weight of prickly pear fruit and 115 parts by weight of distilled water are mixed and pressed into juice by a juicer. The 24 parts by weight of apigenin aqueous solution, 10 parts by weight of apigenin and 14 parts by weight of active water are mixed and stirred into a solution.

9. The prickly pear fermentation broth that promotes telomerase activity according to claim 1, characterized in that: The 225 parts by weight of prickly pear juice, 152 parts by weight of prickly pear fruit, and 73 parts by weight of distilled water were mixed and pressed into juice using a juicer. The 140 parts by weight of apigenin aqueous solution, 19 parts by weight of apigenin, and 121 parts by weight of active water were mixed and stirred into a solution.

10. The prickly pear fermentation broth that promotes telomerase activity according to claim 1, characterized in that: The 240 parts by weight of prickly pear juice, 180 parts by weight of prickly pear fruit, and 60 parts by weight of distilled water are mixed and pressed into juice using a juicer. The 150 parts by weight of apigenin aqueous solution, 25 parts by weight of apigenin, and 125 parts by weight of active water are mixed and stirred into a solution.

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