Health-care wine for improving gout, hypertension and antioxidation and its making process

Abstract

The present application belongs to the technical field of brewing, and mainly relates to a health-care wine for improving gout and high blood pressure and an improving process thereof. The present application provides a compound microbial inoculant and a process for preparing the health-care wine. The raw materials of the health-care wine are black beans and wheat, and the process comprises the following steps: crushing the raw materials, high-temperature impregnation, steaming, ingredient mixing, airing, temperature reduction, sugar addition, yeast addition, addition of pre-fermented raspberries, astragalus root, radix angelicae dahuricae, apple peel, pitaya peel, and lotus leaf, fermentation in a jar, termination of fermentation, clarification, and black bean health-care wine is obtained. The black bean health-care wine prepared by the present application has the effects of relieving and preventing gout and hyperuricemia, resisting oxidation and aging, and preventing cardiovascular and cerebrovascular diseases, and has a soft taste and pure flavor, and is the first choice of health-care wine for patients with gout and hyperuricemia. In addition, the black bean health-care wine prepared by the present application has an unexpected blood pressure lowering function.

Description

Technical Field

[0001] This invention belongs to the field of brewing technology, and mainly relates to a black bean wine, specifically a compound microbial agent, a health wine with the functions of improving gout and hypertension and anti-oxidation, and its production process. Background Technology

[0002] Long-term consumption of high-proof liquor can cause vasoconstriction and elevated blood pressure. For patients with unstable blood pressure, this can lead to a sudden spike in blood pressure, potentially causing hypertensive encephalopathy, cerebral hemorrhage, and other complications. Because alcohol can affect uric acid synthesis and metabolism through multiple pathways, drinking alcohol has always been a contraindication for gout patients. Multiple domestic and international gout treatment guidelines explicitly state that gout patients must strictly abstain from all alcoholic beverages. Many patients develop hyperuricemia and gout due to excessive alcohol consumption, but very few can strictly abstain from alcohol. Many patients frequently drink alcohol due to work, social interactions, and lifestyle habits, while some are habitual drinkers or even alcohol addicts. Therefore, developing a health-promoting liquor that can slow down uric acid levels and reduce the frequency of gout attacks can specifically meet the health needs of this group. Summary of the Invention

[0003] To achieve the above objectives, the present invention provides the following technical solution:

[0004] A health tonic for improving gout, hypertension, and providing antioxidant benefits, and its manufacturing process, characterized by:

[0005] The raw materials are black beans and wheat, which are produced through a co-fermentation process with raspberries, astragalus, angelica root, apple peel, dragon fruit peel, and lotus leaves. The process includes: crushing the raw materials, high-temperature soaking, steaming, mixing, spreading and cooling, adding sugar, adding yeast, adding raspberries, astragalus, angelica root, apple peel, dragon fruit peel, and lotus leaves pre-fermented with Clostridium butyricum ATCC19398 (purchased from Shanghai Beinuo Biotechnology Co., Ltd.) and yeast CMCC(B)98007 (purchased from Beijing Bio-Bio Biotechnology Co., Ltd.), fermenting in vats, stopping fermentation, and clarification to obtain black bean health wine.

[0006] Preferably, in the above-mentioned process of making black bean wine with health benefits, the ratio of black beans to wheat is 1:1.5.

[0007] Preferably, in the above-mentioned black bean wine process with health benefits, the ratio of whole black beans to broken black beans is 30:70, and the ratio of whole wheat beans to broken wheat beans is 20:80.

[0008] Preferably, in the above-mentioned process of making black bean wine with health benefits, raspberries, astragalus, angelica root, apple peel, dragon fruit peel, and lotus leaves are added and fermented with black beans.

[0009] Preferably, in the above-mentioned black bean wine process with health benefits, before co-fermentation with black beans, *Clostridium butyricum* ATCC19398 and yeast CMCC(B)98007 are used to pre-ferment raspberries, astragalus, angelica root, apple peel, dragon fruit peel, and lotus leaves.

[0010] Preferably, in the above-mentioned process for making black bean wine with health benefits, the pre-fermented raspberries, astragalus, angelica root, apple peel, dragon fruit peel, and lotus leaves are prepared using the following methods:

[0011] Preparation of Clostridium butyricum ATCC19398 bacterial suspension: The concentration was 1.5 × 10⁻⁶. 9 ~3×10 9 A cfu / ml Clostridium butyricum ATCC19398 bacterial suspension is available for use.

[0012] Preparation of yeast CMCC(B)98007 bacterial culture: The concentration was 3.5 × 10⁻⁶. 9 ~4.6×10 9 Prepare a cfu / ml culture of yeast CMCC(B)98007 for later use;

[0013] Pre-treatment of raspberries, astragalus, angelica root, apple peel, dragon fruit peel and lotus leaf: Wash raspberries, astragalus, angelica root, wash and dry them and then crush them. Wash apple peel, dragon fruit peel and lotus leaf and crush them directly.

[0014] Take a portion of crushed raspberry, astragalus, angelica root, apple peel, dragon fruit peel and lotus leaf and extract them with boiling water. Set aside the extract for later use.

[0015] Pre-fermentation of raspberry, astragalus, angelica root, apple peel, dragon fruit peel, and lotus leaf: Mix 100-200g of crushed raspberry, 50-100g of astragalus, and 50-100g of angelica root evenly, add 100-200ml of Clostridium butyricum ATCC19398 bacterial solution, and ferment at 30-37℃ for 3-5 hours to obtain the pre-fermented product of raspberry, astragalus, and angelica root. Add 100-200g of crushed apple peel... Mix 100-200g of dragon fruit peel and 100-200g of lotus leaves evenly, add 100-200ml of yeast CMCC(B)98007, and ferment at 25-33℃ for 3-5 hours to obtain a pre-fermented product of apple peel, dragon fruit peel, and lotus leaves. Then mix the fermented product of raspberry, astragalus, and angelica root with the pre-fermented product of apple peel, dragon fruit peel, and lotus leaves evenly, and ferment at 32℃ for another 3-5 hours to obtain a mixed fermented product for later use.

[0016] Preferably, in the above-mentioned process for making black bean wine with health benefits, the specific feeding steps are as follows:

[0017] S1. Raw material mixing: Stir-fry the plump and round black beans until they release their aroma, and then mix them with wheat after they cool down naturally. Each batch of raw materials is 150 kg, with a total of 60 kg of black beans (whole grains to broken grains in a ratio of 3:7) and a total of 90 kg of wheat (whole grains to broken grains in a ratio of 2:8).

[0018] S2. High-temperature soaking: Heat the extracts of raspberry, astragalus, angelica root, apple peel, dragon fruit peel and lotus leaf to above 90°C. Add them evenly at a ratio of 45% of the raw material weight. Stir quickly and evenly during the process. Soak for 24 hours, stirring twice during the process to ensure even and thorough soaking.

[0019] S3. Steaming and Ingredient Preparation: Sprinkle 1.5 kg of rice husks at the bottom of the steamer. After steam is generated, load the steamer. Sprinkle the grains when steam appears. After steam is fully generated, cover the steamer and steam the grains over high heat for 2.5 hours. Steam the grains until they are about 70% cooked and have 2-3 parts hard or white core. Then remove them from the steamer.

[0020] S4. Spreading, cooling, adding sugar, adding yeast: Take the pre-treated raw sand out of the steamer, add 180 kg of 90℃ hot water to kill the live bacteria in the pre-treatment, stir evenly, put it on the cooling machine to cool down to 20℃, add 50 kg of white sugar, stir and mix evenly, and add yeast.

[0021] S5. Add pre-fermented raspberries, astragalus, angelica root, apple peel, dragon fruit peel, and lotus leaves: After mixing white sugar and yeast evenly, add the pre-fermented mixture of raspberries, astragalus, angelica root, apple peel, dragon fruit peel, and lotus leaves. The ratio of raw materials to fermented product is 1:0.2. After adding, stir and mix evenly. When the temperature reaches 20℃, it can be transferred to the fermentation tank. The transfer time should be short to prevent contamination by other bacteria and maintain the normal fermentation temperature.

[0022] S6. Fermentation in tanks: The well-mixed raw materials and pre-fermented materials are placed into fermentation tanks for sealed fermentation; the fermentation period is 7 days, the fermentation temperature is 20℃, and the expected alcohol content is no more than 15°.

[0023] S7. Termination of fermentation: When the fermentation mash reaches the expected alcohol content, place the fermentation tank at 0°C to forcibly stop fermentation;

[0024] S8. Clarification: The clarification method is carried out by refrigeration at 4°C. After the fermented mash has completely settled, it is centrifuged at 6000r / min. The supernatant is then aseptically filtered to obtain the black bean health wine.

[0025] Preferably, in the above-mentioned black bean wine process with health benefits, the ratio of raspberry, astragalus, angelica root, apple peel, dragon fruit peel, and lotus leaf mixed fermentation liquid to raw materials is 0.2:1.

[0026] The health wine of this invention can be used as a health product or as a medicine.

[0027] Compared with the prior art, the present invention has the following beneficial effects:

[0028] 1. The black beans, astragalus, and raspberries used in this invention produce a large amount of total flavonoids, alkaloids, and small molecule peptides during fermentation, which help relieve and prevent gout and hyperuricemia.

[0029] 2. The Astragalus membranaceus used in this invention produces more substances that promote blood circulation and remove blood stasis during the fermentation process.

[0030] 3. The Angelica dahurica root used in this invention can produce substances that moisturize the skin and remove blemishes and pigmentation during the fermentation process.

[0031] 4. The apple peels and dragon fruit used in this invention produce substances with antioxidant and anti-aging properties during the fermentation process.

[0032] 5. The alkaloids produced by the fermentation of lotus leaves used in this invention can lower blood lipids and prevent cardiovascular and cerebrovascular diseases.

[0033] 6. The black bean health wine prepared by this invention has an unexpected blood pressure lowering function. Attached Figure Description

[0034] Figure 1 The total flavonoid content in the wine samples of Example 1, Comparative Example 1, and Comparative Example 2 prepared according to the present invention is shown in the figure.

[0035] Figure 2 The graph shows the content of alkaloids and small molecule peptides in the wine samples of Example 1, Comparative Example 1, and Comparative Example 2 prepared according to the present invention.

[0036] Figure 3 The graph shows the content of polyphenols and anthocyanins in the wine samples of Example 2, Comparative Example 3, and Comparative Example 4 prepared according to the present invention. Detailed Implementation

[0037] The following specific examples illustrate the implementation of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and various details in this specification can also be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention. It should be noted that, unless otherwise specified, the following embodiments and features described therein can be combined with each other.

[0038] Example 1

[0039] S1. Preparation of bacterial suspension: Preparation of Clostridium butyricum ATCC19398 bacterial suspension: The concentration was 3×10⁻⁶. 9Prepare a CFU / ml Clostridium butyricum ATCC19398 bacterial suspension for later use; prepare a Yeast CMCC(B)98007 bacterial suspension with a concentration of 4.6 × 10⁻⁶. 9 Yeast culture CMCC(B)98007 was prepared for use.

[0040] S2. Pre-treatment of raspberries, astragalus, angelica root, apple peel, dragon fruit peel and lotus leaf: Wash raspberries, astragalus, angelica root, wash and dry them and then crush them. Wash apple peel, dragon fruit peel and lotus leaf and crush them directly.

[0041] S3. Preparation of extract and fermentation product: (1) Preparation of extract: Take a portion of the crushed raspberry, astragalus, angelica root, apple peel, dragon fruit peel and lotus leaf from step S2 and extract with boiling water. The extract obtained after extraction is ready for use; (2) Pre-fermentation of raspberry, astragalus, angelica root, apple peel, dragon fruit peel and lotus leaf: Mix 200g of crushed raspberry, 100g of astragalus and 100g of angelica root evenly, add 200ml of Clostridium butyricum ATCC19398 bacterial solution and heat at 300ml. Ferment at 3℃ for 5 hours to obtain a pre-fermented product of raspberry, astragalus, and angelica root. Mix 200g of crushed apple peel, 200g of dragon fruit peel, and 200g of lotus leaf evenly, add 200ml of yeast CMCC(B)98007 solution, and ferment at 30℃ for 5 hours to obtain a pre-fermented product of apple peel, dragon fruit peel, and lotus leaf. Then mix the fermented product of raspberry, astragalus, and angelica root with the apple peel, dragon fruit peel, and lotus leaf evenly and ferment at 32℃ for another 5 hours to obtain a mixed fermented product for later use.

[0042] S4. Raw material mixing: Stir-fry the plump and round black beans until they release their aroma, and then mix them with wheat after they cool down naturally. Each batch of raw materials is 150 kg, with a total of 60 kg of black beans (whole grains to broken grains in a ratio of 3:7) and a total of 90 kg of wheat (whole grains to broken grains in a ratio of 2:8).

[0043] S5. High-temperature soaking: Heat the extracts of raspberry, astragalus, angelica root, apple peel, dragon fruit peel and lotus leaf to above 90°C. Add them evenly at a ratio of 45% of the raw material weight. Stir quickly and evenly during the process. Soak for 24 hours, stirring twice during the process to ensure even and thorough soaking.

[0044] S6. Steaming and Ingredient Preparation: Sprinkle 1.5 kg of rice husks at the bottom of the steamer. After steam is generated, load the steamer. Sprinkle the grain when steam appears. After steam is generated, cover the steamer and steam the grain over high heat for 2.5 hours. Steam the grain until it is about 70% cooked and contains 20% hard or white core. Then it can be taken out of the steamer.

[0045] S7. Spreading, cooling, adding sugar, adding yeast: Take the pre-treated raw sand out of the steamer, add 180 kg of 90℃ hot water to kill the live bacteria in the pre-treatment, stir evenly, put it on the cooling machine to cool down to 20℃, add 50 kg of white sugar, stir and mix evenly, and add yeast.

[0046] S8. Add pre-fermented raspberries, astragalus, angelica root, apple peel, dragon fruit peel, and lotus leaves: After mixing white sugar and yeast evenly, add the pre-fermented mixture of raspberries, astragalus, angelica root, apple peel, dragon fruit peel, and lotus leaves. The ratio of raw materials to fermented product is 1:0.2. After adding, stir and mix evenly. When the temperature reaches 20℃, it can be transferred to the fermentation tank. The transfer time should be short to prevent contamination by other bacteria and maintain the normal fermentation temperature.

[0047] S9. Fermentation in tanks: The well-mixed raw materials and pre-fermented materials are placed into fermentation tanks for sealed fermentation; the fermentation period is 7 days, the fermentation temperature is 20℃, and the expected alcohol content is no more than 15°.

[0048] S10. Termination of fermentation: When the fermentation mash reaches the expected alcohol content, place the fermentation tank at 0°C to forcibly stop fermentation.

[0049] S11. Clarification: The clarification method is carried out by refrigeration at 4°C. After the fermented mash has completely settled, it is centrifuged at 6000r / min. The supernatant is then aseptically filtered to obtain the black bean health wine.

[0050] Comparative Example 1: Except for the absence of Astragalus membranaceus in step S2, all other steps were the same as in Example 1.

[0051] Comparative Example 2: Except for step S2, which does not include raspberries, the rest of the steps are the same as in Example 1.

[0052] The prepared black bean health wine was subjected to component analysis. The total flavonoids were analyzed using the method described in (Qiao Zian. Identification of flavonoids in sunflower discs and study on preparation process of sunflower disc granules [D]. Jilin: Jilin University, 2021.); the total alkaloids were analyzed using the method described in (Liu Xiaobo, Xue Junlai, Zhao Haifeng. Comparative analysis of effective component content of sunflower discs in Taonan and other regions [J]. Family Medicine. Medical Treatment and Drug Selection, 2017(07):124-125.); and the small molecule peptides were analyzed using the method described in (Guo Baosheng, Yang Yiyan, Hu Junhan, Han Weiwei, Li Wannan. Optimization of enzyme hydrolysis method for preparing small molecule peptides from sunflower discs using response surface methodology [J]. Journal of Jilin University (Science Edition), 2019, 57(05):1275-1280.). For polyphenol detection methods, refer to (Han Xue. Determination of polyphenol content and fingerprint analysis of green fruit medicinal materials [D]. Jinzhou Medical University, 2017.); for anthocyanin detection methods, refer to (Yang Yang. Analysis of proanthocyanidin content in black goji berry fruits from different sources [D]. Inner Mongolia Agricultural University, 2018.).

[0053] like Figure 1 The image shows the total flavonoid content in the wine samples of Example 1, Comparative Example 1, and Comparative Example 2 prepared according to the present invention. Figure 1 It can be seen that the total flavonoid content decreases significantly when fermentation is carried out without the addition of astragalus. However, the absence of raspberry has little effect on the total flavonoid content. This indicates that the simultaneous addition of black beans and astragalus can significantly increase the total flavonoid content. Figure 2 The graph shows the content of alkaloids and small molecule peptides in the wine samples of Example 1, Comparative Example 1, and Comparative Example 2 prepared according to the present invention. Figure 2 It is known that fermentation without the addition of astragalus and raspberry leads to a significant decrease in the content of alkaloids and small molecule peptides. This indicates that the simultaneous addition of astragalus and raspberry has a certain synergistic effect, which can effectively increase the content of alkaloids and small molecule peptides. The high content of total flavonoids, alkaloids, and small molecule peptides in health wine helps to alleviate and prevent gout and hyperuricemia.

[0054] Example 2

[0055] S1. Preparation of bacterial suspension: Preparation of Clostridium butyricum ATCC19398 bacterial suspension: The concentration was 1.5 × 10⁻⁶. 9 Prepare a CFU / ml Clostridium butyricum ATCC19398 bacterial suspension for later use; prepare a Yeast CMCC(B)98007 bacterial suspension with a concentration of 3.5 × 10⁻⁶. 9 Yeast culture CMCC(B)98007 was prepared for use.

[0056] S2. Pre-treatment of raspberries, astragalus, angelica root, apple peel, dragon fruit peel and lotus leaf: Wash raspberries, astragalus, angelica root, wash and dry them and then crush them. Wash apple peel, dragon fruit peel and lotus leaf and crush them directly.

[0057] S3. Preparation of extract and fermentation product: (1) Preparation of extract: Take a portion of the crushed raspberry, astragalus, angelica root, apple peel, dragon fruit peel and lotus leaf from step S2 and extract with boiling water. The extract obtained after extraction is ready for use. (2) Pre-fermentation of raspberry, astragalus, angelica root, apple peel, dragon fruit peel and lotus leaf: Mix 100g of crushed raspberry, 50g of astragalus and 50g of angelica root evenly, add 100ml of Clostridium butyricum ATCC19398 bacterial solution and ferment at 34℃. Ferment for 3 hours to obtain a pre-fermented product of raspberry, astragalus, and angelica root. Mix 100g of crushed apple peel, 100g of dragon fruit peel, and 100g of lotus leaf evenly, add 100ml of yeast CMCC(B)98007 solution, and ferment at 31℃ for 3 hours to obtain a pre-fermented product of apple peel, dragon fruit peel, and lotus leaf. Mix the pre-fermented product of raspberry, astragalus, and angelica root with the pre-fermented product of apple peel, dragon fruit peel, and lotus leaf evenly, and ferment at 32℃ for another 3 hours. The resulting mixed fermented product is ready for use.

[0058] S4. Raw material mixing: Stir-fry the plump and round black beans until they release their aroma, and then mix them with wheat after they cool down naturally. Each batch of raw materials is 150 kg, with a total of 60 kg of black beans (whole grains to broken grains in a ratio of 3:7) and a total of 90 kg of wheat (whole grains to broken grains in a ratio of 2:8).

[0059] S5. High-temperature soaking: Heat the extracts of raspberry, astragalus, angelica root, apple peel, dragon fruit peel and lotus leaf to above 90°C. Add them evenly at a ratio of 45% of the raw material weight. Stir quickly and evenly during the process. Soak for 24 hours, stirring twice during the process to ensure even and thorough soaking.

[0060] S6. Steaming and Ingredient Preparation: Sprinkle 1.5 kg of rice husks at the bottom of the steamer. After steam is generated, load the steamer. Sprinkle the grain when steam appears. After steam is fully generated, cover the steamer and steam the grain over high heat for 2.5 hours. Steam the grain until it is about 70% cooked and contains 30% hard or white core. Then it can be taken out of the steamer.

[0061] S7. Spreading, cooling, adding sugar, adding yeast: Take the pre-treated raw sand out of the steamer, add 180 kg of 90℃ hot water to kill the live bacteria in the pre-treatment, stir evenly, put it on the cooling machine to cool down to 20℃, add 50 kg of white sugar, stir and mix evenly, and add yeast.

[0062] S8. Add pre-fermented raspberries, astragalus, angelica root, apple peel, dragon fruit peel, and lotus leaves: After mixing white sugar and yeast evenly, add the pre-fermented mixture of raspberries, astragalus, angelica root, apple peel, dragon fruit peel, and lotus leaves. The ratio of raw materials to fermented product is 1:0.2. After adding, stir and mix evenly. When the temperature reaches 20℃, it can be transferred to the fermentation tank. The transfer time should be short to prevent contamination by other bacteria and maintain the normal fermentation temperature.

[0063] S9. Fermentation in tanks: The well-mixed raw materials and pre-fermented materials are placed into fermentation tanks for sealed fermentation; the fermentation period is 7 days, the fermentation temperature is 20℃, and the expected alcohol content is no more than 15°.

[0064] S10. Termination of fermentation: When the fermentation mash reaches the expected alcohol content, place the fermentation tank at 0°C to forcibly stop fermentation.

[0065] S11. Clarification: The clarification method is carried out by refrigeration at 4°C. After the fermented mash has completely settled, it is centrifuged at 6000r / min. The supernatant is then aseptically filtered to obtain the black bean health wine.

[0066] Comparative Example 3: Except for step S3, in which apple peels are not added for fermentation, all other steps are the same as in Example 2.

[0067] Comparative Example 4: Except for step S3, in which dragon fruit peel is not added for fermentation, all other steps are the same as in Example 2.

[0068] like Figure 3 The figures shown are graphs depicting the polyphenol and anthocyanin content in the wine samples of Example 2, Comparative Example 3, and Comparative Example 4 prepared according to the present invention. Figure 3 It can be seen that not fermenting apple peels leads to a significant decrease in the content of antioxidants such as polyphenols and anthocyanins, with a greater impact on polyphenol content. Fermenting dragon fruit peels, on the other hand, results in a sharp decrease in anthocyanin content. This indicates that a specific co-fermentation process using apple and dragon fruit peels can effectively increase the content of antioxidants such as polyphenols and anthocyanins, thus playing an anti-aging role.

[0069] Example 3

[0070] S1. Preparation of bacterial suspension: Preparation of Clostridium butyricum ATCC19398 bacterial suspension: The concentration was 1.5 × 10⁻⁶. 9 Prepare a CFU / ml Clostridium butyricum ATCC19398 bacterial suspension for later use; prepare a Yeast CMCC(B)98007 bacterial suspension with a concentration of 3.5 × 10⁻⁶. 9 Yeast culture CMCC(B)98007 was prepared for use.

[0071] S2. Pre-treatment of raspberries, astragalus, angelica root, apple peel, dragon fruit peel and lotus leaf: Wash raspberries, astragalus, angelica root, wash and dry them and then crush them. Wash apple peel, dragon fruit peel and lotus leaf and crush them directly.

[0072] S3. Preparation of extract and fermentation product: (1) Preparation of extract: Take a portion of the crushed raspberry, astragalus, angelica root, apple peel, dragon fruit peel and lotus leaf from step S2 and extract with boiling water. The extract obtained after extraction is ready for use. (2) Pre-fermentation of raspberry, astragalus, angelica root, apple peel, dragon fruit peel and lotus leaf: Mix 150g of crushed raspberry, 70g of astragalus and 70g of angelica root evenly, add 150ml of Clostridium butyricum ATCC19398 bacterial solution and ferment at 33℃. Ferment for 4 hours to obtain a pre-fermented product of raspberry, astragalus, and angelica root. Mix 150g of crushed apple peel, 150g of dragon fruit peel, and 150g of lotus leaf evenly, add 150ml of yeast CMCC(B)98007 solution, and ferment at 31℃ for 4 hours to obtain a pre-fermented product of apple peel, dragon fruit peel, and lotus leaf. Then mix the pre-fermented product of raspberry, astragalus, and angelica root with the pre-fermented product of apple peel, dragon fruit peel, and lotus leaf evenly, and ferment at 32℃ for another 4 hours to obtain a mixed fermented product for later use.

[0073] S4. Raw material mixing: Stir-fry the plump and round black beans until they release their aroma, and then mix them with wheat after they cool down naturally. Each batch of raw materials is 150 kg, with a total of 60 kg of black beans (whole grains to broken grains in a ratio of 3:7) and a total of 90 kg of wheat (whole grains to broken grains in a ratio of 2:8).

[0074] S5. High-temperature soaking: Heat the extracts of raspberry, astragalus, angelica root, apple peel, dragon fruit peel and lotus leaf to above 90°C. Add them evenly at a ratio of 45% of the raw material weight. Stir quickly and evenly during the process. Soak for 24 hours, stirring twice during the process to ensure even and thorough soaking.

[0075] S6. Steaming and Ingredient Preparation: Sprinkle 1.5 kg of rice husks at the bottom of the steamer. After steam is generated, load the steamer. Sprinkle the grain when steam appears. After steam is fully generated, cover the steamer and steam the grain over high heat for 2.5 hours. Steam the grain until it is about 70% cooked and contains 30% hard or white core. Then it can be taken out of the steamer.

[0076] S7. Spreading, cooling, adding sugar, adding yeast: Take the pre-treated raw sand out of the steamer, add 180 kg of 90℃ hot water to kill the live bacteria in the pre-treatment, stir evenly, put it on the cooling machine to cool down to 20℃, add 50 kg of white sugar, stir and mix evenly, and add yeast.

[0077] S8. Add pre-fermented raspberries, astragalus, angelica root, apple peel, dragon fruit peel, and lotus leaves: After mixing white sugar and yeast evenly, add the pre-fermented mixture of raspberries, astragalus, angelica root, apple peel, dragon fruit peel, and lotus leaves. The ratio of raw materials to fermented product is 1:0.2. After adding, stir and mix evenly. When the temperature reaches 20℃, it can be transferred to the fermentation tank. The transfer time should be short to prevent contamination by other bacteria and maintain the normal fermentation temperature.

[0078] S9. Fermentation in tanks: The well-mixed raw materials and pre-fermented materials are placed into fermentation tanks for sealed fermentation; the fermentation period is 7 days, the fermentation temperature is 20℃, and the expected alcohol content is no more than 15°.

[0079] S10. Termination of fermentation: When the fermentation mash reaches the expected alcohol content, place the fermentation tank at 0°C to forcibly stop fermentation.

[0080] S11. Clarification: The clarification method is carried out by refrigeration at 4°C. After the fermented mash has completely settled, it is centrifuged at 6000r / min. The supernatant is then aseptically filtered to obtain the black bean health wine.

[0081] Comparative Example 5: Except for step S3, which does not use Clostridium butyricum ATCC19398 bacterial culture for fermentation, all other steps are the same as in Example 3.

[0082] Comparative Example 6: Except for step S3, in which yeast CMCC(B)98007 was not used for fermentation, all other steps were the same as in Example 3.

[0083] Comparative Example 7: Except for replacing Clostridium butyricum ATCC19398 in step S3 with Clostridium butyricum CGMCC 1.5025 (purchased from Shanghai Luhui Technology Co., Ltd.), everything else is the same as in Example 3.

[0084] Comparative Example 8: Except for replacing yeast CMCC(B)98007 in step S3 with yeast ATCC 208298 (purchased from BioWind Resources), everything else is the same as in Example 3.

[0085] Table 1 shows that the absence of *Clostridium butyricum* ATCC19398 or *C. butyricum* CMCC(B)98007 significantly reduced the content of total flavonoids, alkaloids, small peptides, polyphenols, and anthocyanins in the wine samples. This indicates that the simultaneous use of *Clostridium butyricum* ATCC19398 and *C. butyricum* CMCC(B)98007 during pre-fermentation can exert an unexpected synergistic effect, making it easier to obtain total flavonoids, alkaloids, small peptides, polyphenols, and anthocyanins in the fermented product. Meanwhile, Comparative Examples 7 and 8 show that *Clostridium butyricum* ATCC19398 or the absence of *C. butyricum* CMCC(B)98007 exhibits specificity for black bean wine fermentation.

[0086] Table 1 Content of various substances in wine samples

[0087]

[0088]

[0089] Example 4

[0090] S1. Preparation of bacterial suspension: Preparation of Clostridium butyricum ATCC19398 bacterial suspension: The concentration was 2×10⁻⁶. 9 Prepare a CFU / ml Clostridium butyricum ATCC19398 bacterial suspension for later use; prepare a Yeast CMCC(B)98007 bacterial suspension with a concentration of 4.3 × 10⁻⁶. 9 Yeast culture CMCC(B)98007 was prepared for use.

[0091] S2. Pre-treatment of raspberries, astragalus, angelica root, apple peel, dragon fruit peel and lotus leaf: Wash raspberries, astragalus, angelica root, wash and dry them and then crush them. Wash apple peel, dragon fruit peel and lotus leaf and crush them directly.

[0092] S3. Preparation of extract and fermentation product: (1) Preparation of extract: Take a portion of the crushed raspberry, astragalus, angelica root, apple peel, dragon fruit peel and lotus leaf from step S2 and extract with boiling water. The extract obtained after extraction is ready for use. (2) Pre-fermentation of raspberry, astragalus, angelica root, apple peel, dragon fruit peel and lotus leaf: Mix 170g of crushed raspberry, 90g of astragalus and 90g of angelica root evenly, add 170ml of Clostridium butyricum ATCC19398 bacterial solution and ferment at 33℃. Ferment for 4 hours to obtain a pre-fermented product of raspberry, astragalus, and angelica root. Mix 170g of crushed apple peel, 170g of dragon fruit peel, and 170g of lotus leaf evenly, add 170ml of yeast CMCC(B)98007 solution, and ferment at 32℃ for 4 hours to obtain a pre-fermented product of apple peel, dragon fruit peel, and lotus leaf. Then mix the pre-fermented product of raspberry, astragalus, and angelica root with the pre-fermented product of apple peel, dragon fruit peel, and lotus leaf evenly, and ferment at 32℃ for another 4 hours to obtain a mixed fermented product for later use.

[0093] S4. Raw material mixing: Stir-fry the plump and round black beans until they release their aroma, and then mix them with wheat after they cool down naturally. Each batch of raw materials is 150 kg, with a total of 60 kg of black beans (whole grains to broken grains in a ratio of 3:7) and a total of 90 kg of wheat (whole grains to broken grains in a ratio of 2:8).

[0094] S5. High-temperature soaking: Heat the extracts of raspberry, astragalus, angelica root, apple peel, dragon fruit peel and lotus leaf to above 90°C. Add them evenly at a ratio of 45% of the raw material weight. Stir quickly and evenly during the process. Soak for 24 hours, stirring twice during the process to ensure even and thorough soaking.

[0095] S6. Steaming and Ingredient Preparation: Sprinkle 1.5 kg of rice husks at the bottom of the steamer. After steam is generated, load the steamer. Sprinkle the grain when steam appears. After steam is generated, cover the steamer and steam the grain over high heat for 2.5 hours. Steam the grain until it is about 70% cooked and contains 20% hard or white core. Then it can be taken out of the steamer.

[0096] S7. Spreading, cooling, adding sugar, adding yeast: Take the pre-treated raw sand out of the steamer, add 180 kg of 90℃ hot water to kill the live bacteria in the pre-treatment, stir evenly, put it on the cooling machine to cool down to 20℃, add 50 kg of white sugar, stir and mix evenly, and add yeast.

[0097] S8. Add pre-fermented raspberries, astragalus, angelica root, apple peel, dragon fruit peel, and lotus leaves: After mixing white sugar and yeast evenly, add the pre-fermented mixture of raspberries, astragalus, angelica root, apple peel, dragon fruit peel, and lotus leaves. The ratio of raw materials to fermented product is 1:0.2. After adding, stir and mix evenly. When the temperature reaches 20℃, it can be transferred to the fermentation tank. The transfer time should be short to prevent contamination by other bacteria and maintain the normal fermentation temperature.

[0098] S9. Fermentation in tanks: The well-mixed raw materials and pre-fermented materials are placed into fermentation tanks for sealed fermentation; the fermentation period is 7 days, the fermentation temperature is 20℃, and the expected alcohol content is no more than 15°.

[0099] S10. Termination of fermentation: When the fermentation mash reaches the expected alcohol content, place the fermentation tank at 0°C to forcibly stop fermentation.

[0100] S11. Clarification: The clarification method is carried out by refrigeration at 4°C. After the fermented mash has completely settled, it is centrifuged at 6000r / min. The supernatant is then aseptically filtered to obtain the black bean health wine.

[0101] Comparative Example 9: Except for replacing the impregnation solution with purified water in step S5, the rest is the same as in Example 4.

[0102] According to the scoring criteria in Table 2, using extracts of raspberry, astragalus, angelica root, apple peel, dragon fruit peel, and lotus leaf as the high-temperature maceration solution can produce black bean health wine with uniform color, no obvious suspension or sediment, good aroma, and a smooth and pure taste.

[0103] Table 2 Scoring Criteria for Health Tofu

[0104]

[0105] Table 3. Wine sample scores for Example 1 and Comparative Example 9

[0106] Wine sample Score Color Organizational status odor taste Example 4 98 20 20 29 29 Comparative Example 9 56 16 16 12 12

[0107] Example 5: Blood Pressure Lowering Effect Test

[0108] Animal experiments

[0109] Fifty-five 13-week-old male C57BL / 6J rats (purchased in Beijing), weighing 230–250 g, were acclimatized in an SPF-grade animal facility under the following conditions: free access to food and water, temperature maintained at 23℃–25℃, humidity at 55% ± 5%, and a 12-hour diurnal cycle. After one week of acclimatization, five rats were randomly selected as the control group and fed normally. The remaining rats were used to establish a rat model. Starting from day 8, the rats in the model group were administered L-nitroarginine (L-NA) by gavage at 9:00 AM daily at a dose of 490 mg / kg / day, while the control group rats were administered an equal volume of physiological saline by gavage. The model was maintained for 28 consecutive days. During the gavage period, at 15:00 on the 7th, 14th, 21st and 28th day of modeling, the blood pressure of the rats in the modeling group was measured using the BP-98A non-invasive rat blood pressure measurement system. The systolic blood pressure value of the rat tail was measured. Each rat was measured three times and the average value was calculated. When the average systolic blood pressure was ≥180 mmHg, the modeling was considered successful.

[0110] Rats that successfully developed the model were randomly divided into a model group, an example group (four groups: Examples 1-4), a control group (four groups: Comparative Examples 5-8), and a positive control group, with five rats in each group. The example groups were administered the black bean health wine prepared in Examples 1-4 by gavage at a dose of 0.1 ml / 10 g body weight daily. The control groups were administered the black bean health wine prepared in Comparative Examples 5-8 by gavage at a dose of 0.1 ml / 10 g body weight daily. The positive control group was administered commercially available health wine with an alcohol content not exceeding 15° by gavage daily. The normal group and the model group were administered an equal volume of physiological saline by gavage. All treatments were administered once daily for eight consecutive weeks. During the treatment period, rats' body weight, food intake, heart rate, mental state, and activity level were observed and recorded regularly. Blood pressure changes after single and continuous administration were also recorded. For single administration, systolic and diastolic blood pressure were measured before administration and at 30, 60, 90, 120, 150, 180, and 210 minutes after administration. For continuous administration, systolic and diastolic blood pressure were measured before administration and after 60 days of continuous administration, with a measurement interval of 5 days, using the same method as above.

[0111] As shown in Table 4, the black bean health wine prepared in this invention has an unexpected blood pressure lowering function. Compared with the positive control group, the blood pressure of rats treated with the black bean health wine prepared in Examples 1-4 returned to normal after 60 days of intervention, while the commercially available health wines used do not have this function. Furthermore, the black bean health wine obtained by co-fermentation of Clostridium butyricum ATCC19398 and yeast CMCC(B)98007 has an even better blood pressure lowering effect.

[0112] Table 4

[0113]

[0114] The above-described embodiments are merely illustrative of specific implementations of the present invention, and while the descriptions are detailed, they should not be construed as limiting the scope of protection of the present invention. It should be noted that for those skilled in the art, any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention should be considered equivalent substitutions and are included within the scope of protection of the present invention.

Claims

1. A compound microbial agent used for fermenting black bean wine, characterized in that: This includes Clostridium butyricum ATCC19398 and yeast CMCC(B)98007.

2. A method for preparing health wine using the compound microbial agent according to claim 1, characterized in that: The raw materials are black beans and wheat, which are produced through a co-fermentation process with raspberries, astragalus, angelica root, apple peel, dragon fruit peel, and lotus leaves. The process includes: crushing the raw materials, high-temperature soaking, steaming, mixing, spreading and cooling, adding sugar, adding yeast, adding raspberries, astragalus, angelica root, apple peel, dragon fruit peel, and lotus leaves that have been pre-fermented using Clostridium butyricum ATCC19398 and yeast CMCC(B)98007, fermenting in vats, stopping fermentation, and clarifying to obtain black bean health wine.

3. The method according to claim 2, characterized in that: The ratio of black beans to wheat used is 1:1.

5.

4. The method according to claim 2, characterized in that: The ratio of whole to broken black beans used was 30:70, and the ratio of whole to broken wheat beans was 20:

80.

5. The method according to claim 2, characterized in that: Before co-fermentation with black beans, raspberries, astragalus, angelica root, apple peel, dragon fruit peel, and lotus leaves were pre-fermented using Clostridium butyricum ATCC19398 and yeast CMCC(B)98007.

6. The method according to claim 2, characterized in that: The specific feeding steps are as follows: S1. Raw material mixing: Stir-fry the plump and round black beans until they release their aroma, and then mix them with wheat after they cool down naturally. Each batch of raw materials is 150 kg, with a total of 60 kg of black beans (whole grains to broken grains in a ratio of 3:7) and a total of 90 kg of wheat (whole grains to broken grains in a ratio of 2:8). S2. High-temperature soaking: Heat the extracts of raspberry, astragalus, angelica root, apple peel, dragon fruit peel and lotus leaf to above 90°C. Add them evenly at a ratio of 45% of the raw material weight. Stir quickly and evenly during the process. Soak for 24 hours, stirring twice during the process to ensure even and thorough soaking. S3. Steaming and Ingredient Preparation: Sprinkle 1.5 kg of rice husks at the bottom of the steamer. After steam is generated, load the steamer. Sprinkle the grains when steam appears. After steam is fully generated, cover the steamer and steam the grains over high heat for 2.5 hours. Steam the grains until they are about 70% cooked and have 2-3 parts hard or white core. Then remove them from the steamer. S4. Spreading, cooling, adding sugar, adding yeast: Remove the steamed raw sand from the steamer, add 180 kg of 90℃ hot water to kill the pre-treated live bacteria, stir evenly, put it on a cooling machine to cool down to 20℃, add 50 kg of white sugar, stir and mix evenly, and add yeast. S5. Add pre-fermented raspberries, astragalus, angelica root, apple peel, dragon fruit peel, and lotus leaves: After mixing white sugar and yeast evenly, add the pre-fermented mixture of raspberries, astragalus, angelica root, apple peel, dragon fruit peel, and lotus leaves. The ratio of raw materials to fermented mixture is 1:0.

2. After adding, stir and mix evenly. When the temperature reaches 20℃, it can be transferred to the fermentation tank. The transfer time should be short to prevent contamination by other bacteria and maintain the normal fermentation temperature. S6. Fermentation in tanks: The well-mixed raw materials and pre-fermented materials are placed into fermentation tanks for sealed fermentation; the fermentation period is 7 days, the fermentation temperature is 7℃, and the expected alcohol content is no more than 15°. S7. Termination of fermentation: When the fermentation mash reaches the expected alcohol content, place the fermentation tank at 0°C to forcibly stop fermentation; S8. Clarification: The clarification method is carried out by refrigeration at 4℃. After the fermented mash has completely settled, it is centrifuged at 6000r / min. The supernatant is then aseptically filtered to obtain the black bean health wine.

Images