High-activity probiotic soybean clear beer and preparation method thereof

Abstract

The invention relates to the technical field of beer preparation, in particular to high-activity probiotic soybean whey beer and a preparation method thereof, and aims to increase the viable count of probiotics in beer. The preparation method comprises the following steps: fermenting lactic acid bacteria in soybean whey to prepare soybean whey probiotic bacteria base liquid, performing maltolization and boiling wort; and carrying out primary fermentation on the wort, adding the soybean whey and the soybean whey probiotic base solution, and carrying out secondary fermentation to obtain the soybean whey craft beer containing high-activity probiotics. According to the invention, the competitive inhibition effect of saccharomycetes on lactic acid bacteria is reduced through the domestication growth and activity protection effect of the soybean whey on the lactic acid bacteria, so that the activity of probiotics in the beer is fully retained, and the beer has a fermented mellow flavor, also contains rich viable bacteria nutrition and active substances such as leucine, raffinose and stachyose, and has a good taste. The effect of relieving the burdens after drinking is exerted.

Description

Technical Field

[0001] This invention belongs to the technical field of beer fermentation methods, specifically relating to a highly active probiotic soybean whey beer and its preparation method. Background Technology

[0002] Highly active probiotic beverages, with their unique lactic acid bacteria strains and fermentation flavor characteristics, are favored by various consumers and generate good economic benefits. However, craft beer is made through yeast fermentation. The competitive inhibition between lactic acid bacteria and brewer's yeast due to species differences leads to defects such as low viable lactic acid bacteria counts and bland fermentation flavor in probiotic craft beer, limiting its further application. Therefore, developing a substrate that promotes the growth and protects the activity of lactic acid bacteria, reducing the competitive inhibition effect of yeast on lactic acid bacteria during beer brewing, and achieving the preparation of highly active probiotic beer can not only enhance its nutrition and flavor but also meet consumers' demand for a healthy and less burdensome beer.

[0003] Soybean whey is a liquid product separated during soybean product processing, obtained by coagulating (or precipitating) soybean protein using magnesium chloride (brine) and other methods. It is rich in nutrients and is also known as soybean whey or soybean yellow liquid. Currently, soybean whey is generally treated as a cheap liquid waste, wasting its nutritional resources. Turning soybean whey into a valuable resource with significant economic and social value is of great importance. Research shows that soybean whey is rich in nutrients such as soybean polypeptides, soybean oligosaccharides, soybean isoflavones, and minerals, making it an excellent liquid nutrient substrate for the cultivation and proliferation of microorganisms such as lactic acid bacteria. However, current research on soybean whey focuses on the growth and reproduction of lactic acid bacteria, with limited research on its protective effects on lactic acid bacteria activity and its symbiotic relationship with yeast.

[0004] Existing patents and literature show that invention patent CN115011420A uses soybean whey as raw material, and enriches active substances through fermentation with yeast and various probiotics to produce a craft beer with liver-protecting effects. However, this beer has a low number of live bacteria and a high octanoic acid content, affecting the sensory quality of the beer. Invention patent CN112592780A uses soybean whey after ultrafiltration as a brewing raw material for beer. This not only causes the loss of nutrients in the whey, but ultrafiltration also increases energy consumption and production costs. In addition, this beer does not involve the addition of probiotics or their activity preservation. Invention patent CN116286226A increases the content of *Saccharomyces boulardii* probiotics in beer based on a mixed dual-yeast synergistic fermentation mode. Invention patent CN116144543A develops functionally active GR-5 lactic acid bacteria for craft beer. However, the above studies only focus on the role of strain interaction in the nutritional efficacy of beer, without analyzing the application effect of soybean whey adjuvants in the beer strain fermentation process. At the same time, the differences in the effects of different beer brewing adjuvants and processes on the proliferation and metabolism of probiotics during fermentation will cause significant changes in the maintenance of probiotic activity and flavor enhancement, thereby affecting its nutritional and sensory quality. Furthermore, traditional beer and existing whey beer cannot solve the problems of bland flavor and the physical burden such as electrolyte imbalance, diarrhea, and fatigue after drinking. Summary of the Invention

[0005] To address the problems of low probiotic viable counts, bland and acidic flavor, and long fermentation time in existing soy whey beer technology, this invention provides a highly active probiotic soy whey craft beer and its preparation method. Through the domestication and activity protection effects of soy whey on lactic acid bacteria, the competitive inhibition effect of yeast on lactic acid bacteria is reduced during the secondary fermentation of beer, forming a symbiotic state. This promotes the protection and high activity of lactic acid bacteria in the beer, shortens the fermentation time, and enriches nutrients such as leucine, raffinose, and stachyose. The resulting soy whey beer can alleviate the burden on the body after drinking while possessing a rich fermented flavor.

[0006] The technical solution adopted by this invention to solve its technical problem is:

[0007] According to a first aspect of the present invention, a highly active probiotic soybean whey beer is provided, comprising the following components in parts by weight: 8000-10000 parts soybean whey, 8000-10000 parts pure water, 1000-2000 parts barley malt, 800-1000 parts wheat malt, 100-200 parts sugar, 6-10 parts hops, 1-2 parts neutral protease, 5-15 parts yeast, and 1-3 parts lactic acid bacteria.

[0008] According to an embodiment of the present invention, the yeast is Munich brewer's yeast; The hops mentioned are Magnum hops.

[0009] According to an embodiment of the present invention, the soybean whey is a type of soybean whey obtained during the processing of soybean products by coagulating or precipitating soybean protein using salt, acid, and microbial fermentation, and then separating it. The soybean whey can be any one or more of the liquid products separated from soybean protein after coagulation (or precipitation) during the production of tofu, dried tofu, etc.

[0010] According to an embodiment of the present invention, the malt includes one or more of light ale malt and Pilsner malt. The malt used in this invention was purchased from the French Omega Group.

[0011] According to an embodiment of the present invention, both the barley malt and wheat malt are fresh, dry, and free from pests.

[0012] Preferably, the mass ratio of barley malt to wheat malt is 1:1, 1.5:1, or 2:1; when used, they are mixed evenly in proportion to obtain mixed malt.

[0013] More preferably, the mass ratio of barley malt to wheat malt is 1.5:1.

[0014] According to an embodiment of the present invention, the sugar is selected from any one or more of sucrose, granulated sugar, and white sugar.

[0015] According to an embodiment of the present invention, the lactic acid bacteria are one or more of Lactobacillus plantarum and Streptococcus thermophilus.

[0016] According to an embodiment of the present invention, the lactic acid bacteria strains used in the present invention were purchased from Shandong Zhongke Jiayi Bioengineering Co., Ltd., and their serial numbers are: Lactobacillus plantarum 376, Lactobacillus plantarum 002, and Streptococcus thermophilus STN.

[0017] According to an embodiment of the present invention, the number of live probiotics in the highly active probiotic soybean whey beer reaches 1×10⁻⁶. 7 CFU / mL.

[0018] The highly active probiotic soybean whey beer prepared by this invention not only has a live probiotic count of 1×10⁻⁶ bacteria in the beer, but also... 7 With a concentration of CFU / mL and a rich bean aroma, it meets the health and taste needs of beer drinkers, generating economic and social benefits.

[0019] According to a second aspect of the present invention, a method for preparing highly active probiotic soybean whey beer is provided, comprising the following specific steps: Preparation of soybean probiotic base liquid: Boil 300-400 parts of soybean protein and cool to 30-40℃ to obtain soybean protein base liquid; After the soybean protein base solution was hydrolyzed with neutral protease for 4-6 hours, 10 parts of sugar were added, and the solution was sterilized at 105°C and then cooled to 20°C to obtain soybean protein culture medium. The bacterial solution containing lactic acid bacteria has a viable count > 1 × 10⁻⁶. 7 CFU / mL was added to the soybean culture medium and cultured at 37℃ for 8-12 h to prepare soybean probiotic culture medium. Soaking and saccharification: Add heated pure water to the mixture of ground barley malt and wheat malt, and saccharify at 72-82℃ for 1 hour to obtain wort; Boiling the wort: Boil the wort. At the beginning of boiling, add hops at a ratio of 3 / 6 to 4 / 6 by weight. After boiling for 30 to 40 minutes, add the remaining hops and boil for another 30 to 40 minutes to obtain hop wort.

[0020] Primary fermentation: Cool the hop wort to room temperature, add yeast and ferment at 20°C for 6-8 days to obtain primary fermentation liquid; Secondary fermentation: Boil the remaining fresh soybean whey and cool it to room temperature, then mix it with the soybean whey probiotic base liquid, add the remaining sugar (90-190 parts), stir well to obtain soybean whey fermentation liquid, add the soybean whey fermentation liquid to the primary fermentation liquid, and continue fermentation at 20℃ for 6-8 days to obtain the highly active probiotic soybean whey beer.

[0021] Refrigeration: The highly active probiotic soybean beer is refrigerated at 4℃ for 24 hours to stabilize the beer quality during post-maturation.

[0022] During the later stages of primary fermentation in beer, nutrient loss leads to a slowdown in yeast growth and metabolism. Therefore, soybean spore-based fermentation solution containing lactic acid bacteria is added during secondary fermentation to reduce the competitive inhibition of lactic acid bacteria by yeast, promoting the retention and high activity of lactic acid bacteria. Under normal fermentation conditions, *Saccharomyces cerevisiae* rapidly becomes dominant, inhibiting the growth of putrefactive Enterobacteriaceae, lactic acid bacteria, and other competing microorganisms by increasing alcohol concentration, lowering pH, and producing carbon dioxide and sulfur dioxide. When *Saccharomyces cerevisiae* is co-cultured with lactic acid bacteria, fermentation is dominated by *Saccharomyces cerevisiae*, and its rapid growth leads to a reduction in the growth of LAB and *Brettanomyces* cells.

[0023] This invention overcomes the above-mentioned technical problems by adopting a pre-acclimatization strategy. Soybean soybeans are hydrolyzed by neutral protease for 4-6 hours to release more available nitrogen sources. LAB cells complete physiological adaptation in the soybean ...

[0024] The method provided by this invention effectively increases the content of pyruvate, which is the end product of glycolysis and a precursor for amino acid synthesis. The domesticated LABs exhibit enhanced metabolic capacity, promoting energy metabolism and amino acid synthesis. Through alanine aminotransferase-mediated transamination, they accept the amino group of glutamate to produce α-ketoglutarate, providing energy and nitrogen to the cells.

[0025] Furthermore, this invention significantly reduces the content of caprylic acid. The domesticated LAB forms a synergistic metabolism with yeast, reducing abnormal fatty acid metabolism and lowering the production of caprylic acid.

[0026] In Example 1 of this invention, the leucine (Leu) content reached 112.38 mg / L, which was significantly higher than that of the comparative example (23-73 mg / L). Leucine is a branched-chain amino acid that can promote the growth of lactic acid bacteria by regulating energy metabolism. Pyruvate, as a three-carbon α-keto acid skeleton, is an important carbon source for leucine synthesis. The metabolic activity of domesticated LAB is enhanced, promoting amino acid synthesis and secretion.

[0027] According to an embodiment of the present invention, the preparation of the bacterial culture includes: inoculating lactic acid bacteria powder into MRS broth and culturing it at 37°C for 24 h; and preparing a second subculture in the same manner to obtain the bacterial culture, with a viable count > 1 × 10⁻⁶. 7 CFU / mL.

[0028] Beneficial effects The present invention provides a highly active probiotic soybean whey beer and its preparation method, which have the following advantages compared with the prior art: The highly active probiotic soybean whey beer provided by this invention, prepared using soybean whey probiotic base liquid, promotes the domestication and growth of lactic acid bacteria and protects their activity. Furthermore, during the secondary fermentation of beer, when yeast growth and metabolism are slowed, it reduces the competitive inhibitory effect of beer yeast on lactic acid bacteria, ensuring the full preservation of probiotic activity in the beer, shortening fermentation time, enhancing the beer's fermentation flavor while enriching nutrients, and alleviating the burden on the body after drinking. It is then added to the beer along with soybean whey during the secondary fermentation process, allowing lactic acid bacteria to grow preferentially and maintain their activity in the nutrient environment of soybean whey. Simultaneously, specific lactic acid bacteria are screened that can co-grow with yeast, reducing the inhibitory effect of yeast on lactic acid bacteria.

[0029] This invention utilizes yeast and lactic acid bacteria base liquid for co-fermentation, shortening fermentation time and improving beer production efficiency.

[0030] The highly active probiotics (lactic acid bacteria and yeast) in the soy beer provided by this invention produce aromatic flavor substances such as esters and alcohols through growth and metabolism, reduce the production of octanoic acid, and enrich nutrients such as amino acids and oligosaccharides, thereby improving the sensory and nutritional quality of the soy beer.

[0031] This invention transforms soybean processing byproduct "soybean extract" into a valuable resource, achieving high-value recycling. Simultaneously, the product combines probiotic functionality, a unique flavor, and post-drinking relief benefits, significantly enriching the beer market's product range and meeting consumers' comprehensive needs for healthy, delicious, and low-calorie drinking. Attached Figure Description

[0032] Figure 1 This is an electronic tongue radar image of different bean curd beers in an embodiment of the present invention.

[0033] Figure 2 This is an electronic nose radar image of different bean curd beers in an embodiment of the present invention.

[0034] Figure 3 This is a graph showing the analysis of monosaccharide and oligosaccharide content in different bean beer samples from embodiments of the present invention. Detailed Implementation

[0035] The present invention will be further described in detail below with reference to specific embodiments, but the implementation of the present invention should not be limited to the scope of the embodiments described.

[0036] Example 1 A highly active probiotic soybean whey beer has the following ingredients: 9 L (9000 parts) soybean whey, 9 L (9000 parts) purified water, 1.50 kg (1500 parts) pale ale malt, 1.00 kg (1000 parts) pale wheat malt, 160 g (160 parts) sucrose, 10 g (10 parts) Munich classic yeast, 2.50 g (2.5 parts) Lactobacillus plantarum 002, 9.00 g (9 parts) hops, and 1.30 g (1.3 parts) neutral protease. The preparation steps are as follows: 1) Preparation of soybean spore probiotic culture base: Boil 300 mL (300 portions) of fresh soybean spores and cool to 30℃. Add 1.30 g of neutral protease and hydrolyze for 4 h. Then add 10 g of white sugar and sterilize at 105℃ for 8 min. Cool to 20℃ and then add Lactobacillus plantarum 002 culture solution with a viable count > 1×10⁻⁶. 7 CFU / mL, incubated at 37℃ for 8 h, and then temporarily stored at 4℃ for later use to prepare soybean probiotic base solution; 2) Pure water treatment: Heat 9 L of pure water to 72℃; 3) Soaking and saccharification: Add the pure water from step 2) to the ground barley and wheat malt, and saccharify at 72°C for 1 hour to extract the wort; 4) Boil the wort: Boil the wort from step 3) for 60 minutes; First hop addition time: at the start of boiling, 6.00 g (4 / 6) of hops were added; Second hop addition time: 30 min after boiling, 3.00 g (2 / 6) of hops were added; Hop variety: Magnum. 5) Primary fermentation: The wort from step 4) was vortexed and allowed to stand for 15 minutes, cooled to room temperature, siphoned into a fermentation tank, yeast was added and fermented at 20℃ for 6 days to obtain the primary fermentation liquid; 6) Soybean bleach pretreatment: Boil 8.7 L of fresh soybean bleach and then cool to room temperature; 7) Secondary fermentation: Add the soybean whey from step 6) to the primary fermentation liquid from step 5), add 150 g of sugar, add the soybean whey probiotic base liquid from step 1), and continue fermentation at 20℃ for 6 days to obtain highly active probiotic soybean whey beer. 8) Refrigeration: Place the fermented high-activity probiotic soybean beer from step 7) in a refrigerator at 4℃ for 24 hours to mature.

[0037] Example 2 A highly active probiotic soybean whey beer has the following ingredients: 8 L soybean whey, 10 L purified water, 2.00 kg pale ale malt, 0.80 kg pale wheat malt, 150 g sucrose, 12 g Munich classic yeast, 3.00 g Lactobacillus plantarum 376, 6.00 g hops, and 1.00 g neutral protease. The preparation steps are as follows: 1) Preparation of soybean soy protein probiotic base solution: Boil 300 mL of fresh soybean soy protein and cool it to 40℃. Add 1.00 g of neutral protease and hydrolyze for 4 h. Then add 10 g of white sugar and sterilize at 105℃ for 8 min. Cool to 20℃ and then add Lactobacillus plantarum 376 bacterial solution. Incubate at 37℃ for 10 h and then store at 4℃ for later use. 2)-5) are the same as in Example 1; 6) Pretreatment: Boil 7.7 L of fresh soybean water and then cool to room temperature; 7) Secondary fermentation: Add the soybean whey from step 6) to the primary fermentation liquid from step 5), add 140 g of sugar, add the soybean whey probiotic base liquid from step 1), and continue fermentation at 20℃ for 6 days; 8) Refrigeration: Place the fermented beer from step 7) at 4°C for 24 hours.

[0038] Example 3 A highly active probiotic soybean whey beer has the following ingredients: 10 L soybean whey, 8 L purified water, 2.00 kg pale ale malt, 0.80 kg pale wheat malt, 150 g sucrose, 8 g Munich classic yeast, 2.50 g STN Streptococcus thermophilus, 10.00 g hops, and 1.50 g neutral protease. The preparation steps are as follows: 1) Preparation of soybean soy protein probiotic base solution: Boil 300 mL of fresh soybean soy protein and cool it to 30-40℃. Add 1.50 g of neutral protease and hydrolyze for 4 h. Then add 10 g of white sugar and sterilize at 105℃ for 8 min. Cool to 20℃ and then add STN thermophilic streptococcus bacterial solution. Incubate at 37℃ for 8 h and then store at 4℃ for later use. 2) Pure water treatment: Heat 8 L of pure water to 72-92℃; 3) Soaking and saccharification: Add the pure water from step 2) to the ground barley and wheat malt, and saccharify at 72°C for 1 hour to extract the wort; 4) Boil the wort: Boil the wort from step 3) for 60 minutes; 1st hop addition time: at the start of boiling, add 6.00 g of hops; 2nd hop addition time: after boiling for 30 minutes, add 4.00 g of hops; Hop variety: Magnum. 5) Primary fermentation: Let the wort from step 4) settle by vortexing for 15 minutes, cool to room temperature, siphon it into a fermentation tank, add yeast for fermentation, and ferment at 20℃ for 7 days; 6) Pretreatment: Boil 8.7 L of fresh soybean water and then cool to room temperature; 7) Secondary fermentation: Add the soybean whey from step 6) to the fermentation liquid from step 5), add 140 g of sugar, add the soybean whey probiotic base liquid from step 1), and continue fermentation at 20℃ for 7 days; 8) Refrigeration: Place the fermented beer from step 7) at 4°C for 24 hours.

[0039] Example 4 A highly active probiotic soybean whey beer has the following ingredients: 9 L soybean whey, 10 L purified water, 2.00 kg pale ale malt, 1.00 kg pale wheat malt, 100 g sucrose, 5 g Munich classic yeast, 0.50 g Lactobacillus plantarum 002, 0.50 g Lactobacillus plantarum 376, 7.00 g hops, and 1.50 g neutral protease. The preparation steps are as follows: 1) Preparation of soybean soy protein probiotic base solution: Boil 400 mL of fresh soybean soy protein and cool it to 30℃. Add 1.50 g of neutral protease and hydrolyze for 6 h. Then add 10 g of white sugar and sterilize at 105℃ for 8 min. Cool to 20℃ and then add the bacterial solution of Lactobacillus plantarum 002 and Lactobacillus plantarum 376. Incubate at 37℃ for 8 h and then store at 4℃ for later use. 2) Pure water treatment: Heat 9 L of pure water to 92℃; 3) Soaking and saccharification: Add the pure water from step 2) to the ground barley and wheat malt, and saccharify at 82°C for 1 hour to extract the wort; 4) Boil the wort: Boil the wort from step 3) for 60 minutes; 1st hop addition time: at the start of boiling, add 4.00 g of hops; 2nd hop addition time: after boiling for 30 minutes, add 3.00 g of hops; Hop variety: Magnum. 5) Primary fermentation: After step 4), the wort was vortexed and allowed to stand for 15 minutes, cooled to room temperature, siphoned into a fermentation tank, yeast was added, and fermentation was carried out at 20℃ for 8 days. 6) Pretreatment: Boil 8.6 L of fresh soybean water and then cool to room temperature; 7) Secondary fermentation: Add the soybean whey from step 6) to the fermentation liquid from step 5), add 90 g of sugar, add the soybean whey probiotic base liquid from step 1), and continue fermentation at 20℃ for 8 days; 8) Refrigeration: Place the fermented beer from step 7) at 4°C for 24 hours.

[0040] Example 5 A highly active probiotic soybean whey beer has the following ingredients: 10 L soybean whey, 9 L purified water, 1.00 kg Pilsner malt, 0.80 kg pale wheat malt, 120 g sucrose, 8 g Munich classic yeast, 0.83 g Lactobacillus plantarum 002, 0.83 g Lactobacillus plantarum 376, 0.83 g STN Streptococcus thermophilus, 7.00 g hops, and 2.00 g neutral protease. The preparation steps are as follows: 1) Preparation of soybean soy protein probiotic base solution: Boil 300 mL of fresh soybean soy protein and cool it to 30-40℃. Add 2.00 g of neutral protease and hydrolyze for 4 h. Then add 10 g of white sugar and sterilize at 105℃ for 8 min. Cool to 20℃ and then add bacterial solutions of Lactobacillus plantarum 002, Lactobacillus plantarum 376 and STN Streptococcus thermophilus. Incubate at 37℃ for 10 h and then store at 4℃ for later use. 2) Pure water treatment: Heat 9 L of pure water to 72-92℃; 3) Soaking and saccharification: Add the pure water from step 2) to the ground barley and wheat malt, and saccharify at 72°C for 1 hour to extract the wort; 4) Boil the wort: Boil the wort from step 3) for 60 minutes; 1st hop addition time: at the start of boiling, add 4.00 g of hops; 2nd hop addition time: after boiling for 30 minutes, add 3.00 g of hops; Hop variety: Magnum. 5) Primary fermentation: Let the wort from step 4) settle by vortexing for 15 minutes, cool to room temperature, siphon it into a fermentation tank, add yeast for fermentation, and ferment at 20℃ for 7 days; 6) Soybean white protein pretreatment: Boil 9.7 L of fresh soybean white protein, add 110 g of sugar, stir to dissolve, and cool to room temperature; 7) Secondary fermentation: Add the soybean extract from step 6) to the soybean extract probiotic base solution from step 1), and add them together to the fermentation liquid from step 5), and continue fermentation at 20℃ for 7 days; 8) Refrigeration: Place the fermented beer from step 7) at 4°C for 24 hours.

[0041] Comparative Example 1 (Existing methods for preparing soy whey beer using mixed probiotics, employing less soy whey and different probiotic combinations) A soybean whey beer fermented with a mixture of three probiotics has the following ingredients: 3 L soybean whey, 3 L purified water, 1.50 kg pale ale malt, 0.10 kg pale wheat malt, 25 g sucrose, 2.50 g Munich classic yeast, 0.83 g Lactobacillus plantarum, 0.83 g Lactobacillus acidophilus, and 0.83 g Lactobacillus paracasei (probiotic manufacturer: Sichuan Fuji), and 5.00 g hops. The preparation steps are as follows: 1) Soybean bleach pretreatment: Collect 3 L of fresh soybean bleach, add 3 L of pure water to make a soybean bleach aqueous solution, and then heat to 78℃; 2) Raw material pretreatment: Select 1.50 kg of light-colored alfalfa malt and 0.10 kg of light-colored wheat malt free from pests, dry them and then crush them into particles of about 2 mm. 3) Soaking and saccharification: Divide the soybean white solution from step 1) at 78℃ into three portions. Add one portion to the mixed solution from step 2) and keep the temperature at 68℃. Soak for 60 minutes to extract sugar. Observe every 30 minutes and adjust the temperature using the soybean white solution from step 1). 4) Filter the wort: Drain the soaked wort and reflux until the wort is clear. Add the remaining soybean water solution from step 1) to the malt in two batches, and drain the wort after each 10-minute standing period. 5) Boil the wort: Boil the wort obtained in step 4). After boiling for 15 minutes, add 4.00 g of Cascade hops. After boiling for 45 minutes, add 1.00 g of Cascade hops. Stop heating when boiling for 60 minutes. 6) Primary fermentation: Cool the product from step 5) to room temperature, take it out and put it into a fermentation tank. Activate the SO4 yeast with wort at room temperature for 10 min before inoculating it into the wort. Ferment at 22℃ for 14 days. 7) Secondary fermentation: Weigh 25 g of sugar and directly add the bacterial powder: Lactobacillus plantarum, Lactobacillus acidophilus and Lactobacillus paracasei to supplement the fermentation broth in step 6), and continue fermentation at 32℃ for 14 days; 8) Refrigeration: Place the fermented beer in a 4℃ cold storage.

[0042] Comparative Example 2 (Existing methods for preparing soy whey beer using mixed probiotics, employing less soy whey and different probiotic combinations) A soybean whey beer fermented with a mixture of three probiotics has the following ingredients: 3 L soybean whey, 3 L purified water, 1.50 kg pale ale malt, 0.10 kg pale wheat malt, 25 g sucrose, 2.50 g Munich classic yeast, 0.83 g Lactobacillus plantarum 002, 0.83 g Lactobacillus acidophilus, and 0.83 g Lactobacillus paracasei (probiotic manufacturer: Zhongke Jiayi), and 5.00 g hops. The preparation steps are as follows: 1) Soybean bleach pretreatment: Collect 3 L of fresh soybean bleach, add 3 L of pure water to make a soybean bleach aqueous solution, and then heat to 78℃; 2) Raw material pretreatment: Select 1.50 kg of light-colored alfalfa malt and 0.10 kg of light-colored wheat malt free from pests, dry them and then crush them into particles of about 2 mm. 3) Soaking and saccharification: Divide the soybean white solution from step 1) at 78℃ into three portions. Add one portion to the mixed solution from step 2) and keep the temperature at 68℃. Soak for 60 minutes to extract sugar. Observe every 30 minutes and adjust the temperature using the soybean white solution from step 1). 4) Filter the wort: Drain the soaked wort and reflux until the wort is clear. Add the remaining soybean water solution from step 1) to the malt in two batches, and drain the wort after each 10-minute standing period. 5) Boil the wort: Boil the wort obtained in step 4). After boiling for 15 minutes, add 4.00 g of Cascade hops. After boiling for 45 minutes, add 1.00 g of Cascade hops. Stop heating when boiling for 60 minutes. 6) Primary fermentation: Cool the product from step 5) to room temperature, take it out and put it into a fermentation tank. Activate the SO4 yeast with wort at room temperature for 10 min before inoculating it into the wort. Ferment at 22℃ for 14 days. 7) Secondary fermentation: Weigh 25 g of sugar and directly add bacterial powder: Lactobacillus plantarum 002, Lactobacillus plantarum 376, and STN Streptococcus thermophilus to the fermentation broth in step 6), and continue fermentation at 32℃ for 14 days; 8) Refrigeration: Place the fermented beer in a 4℃ cold storage.

[0043] Comparative Example 3 (same as Example 1, except that neutral protease and Lactobacillus plantarum were not added) A soybean whey craft beer, with the following ingredients: 9 L soybean whey, 9 L purified water, 1.50 kg pale ale malt, 1.00 kg pale wheat malt, 160 g sucrose, 10 g Munich classic yeast, and 9.00 g hops. The preparation steps are as follows: 1) Pure water treatment: Heat 9 L of pure water to 72℃; 2) Soaking and saccharification: Add the pure water from step 1) to the ground wheat and barley sprouts, saccharify at 72°C for 1 hour, and then extract the wort; 3) Boil the wort: Boil the wort from step 2) for 60 minutes; 1st hop addition time: at the start of boiling, add 6.00 g of hops; 2nd hop addition time: after boiling for 30 minutes, add 3.00 g of hops; Hop variety: Magnum. 4) Primary fermentation: Let the wort from step 3) settle by vortexing for 15 minutes, cool to room temperature, siphon it into a fermentation tank, add yeast for fermentation, and ferment at 20℃ for 6 days; 5) Soybean bleach pretreatment: Boil 9 L of fresh soybean bleach, stir to dissolve, and cool to room temperature; 6) Secondary fermentation: Add the soybean whites from step 5) to the fermentation liquid from step 4), add 160 g of sugar, and continue fermentation at 20℃ for 6 days; 7) Refrigeration: Place the fermented beer from step 6) at 4°C for 24 hours.

[0044] Comparative Example 4 (same as Example 1, except that no neutral protease was added) A common probiotic soybean whey beer has the following ingredients: 9 L soybean whey, 9 L purified water, 1.50 kg pale ale malt, 1.00 kg pale wheat malt, 160 g sucrose, 10 g Munich classic yeast, 2.50 g Lactobacillus plantarum 002, and 9.00 g hops. The preparation steps are as follows: 1) Pure water treatment: Heat 9 L of pure water to 72℃; 2) Soaking and saccharification: Add the pure water from step 1) to the ground wheat and barley sprouts, saccharify at 72°C for 1 hour, and then extract the wort; 3) Boil the wort: Boil the wort from step 2) for 60 minutes; 1st hop addition time: 6.00 g at the start of boiling; 2nd hop addition time: 3.00 g after boiling for 30 minutes; Hop variety: Magnum. 4) Primary fermentation: Let the wort from step 3) settle by vortexing for 15 minutes, cool to room temperature, siphon it into a fermentation tank, add yeast for fermentation, and ferment at 20℃ for 6 days; 5) Pretreatment: Boil 9 L of fresh soybean water for 20 min and then cool to room temperature; 6) Secondary fermentation: Add the soybean whites from step 5) to the fermentation liquid from step 4), add 160 g of sugar, add activated Lactobacillus plantarum 002 bacterial solution, and continue fermentation at 20℃ for 6 days; 7) Refrigeration: Place the fermented beer from step 6) in a refrigerator at 4°C.

[0045] Comparative Example 5 (same as Example 1, except that Lactobacillus plantarum and yeast were fermented simultaneously in one step). A probiotic soybean whey beer, with the following ingredients: 9 L soybean whey, 9 L purified water, 1.50 kg pale ale malt, 1.00 kg pale wheat malt, 160 g sucrose, 10 g Munich classic yeast, 2.50 g Lactobacillus plantarum 002, 9.00 g hops, and 1.30 g neutral protease. The preparation steps are as follows: 1) Preparation of soybean soy protein probiotic base solution: Boil 300 mL of fresh soybean soy protein and cool it to 30-40℃. Add 1.3 g of neutral protease and hydrolyze for 4 h. Then add 10 g of white sugar and sterilize at 105℃ for 8 min. Cool to 20℃ and then add Lactobacillus plantarum 002 bacterial solution. Incubate at 37℃ for 8 h and then store at 4℃ for later use. 2) Pure water treatment: Heat 9 L of pure water to 72-92℃; 3) Soaking and saccharification: Add the pure water from step 2) to the ground barley and wheat malt, and saccharify at 72°C for 1 hour to extract the wort; 4) Boil the wort: Boil the wort from step 3) for 60 minutes; 1st hop addition time: at the start of boiling, add 6.00 g of hops; 2nd hop addition time: after boiling for 30 minutes, add 3.00 g of hops; Hop variety: Magnum. 5) Primary fermentation: Let the wort from step 4) settle by vortexing for 15 minutes, cool to room temperature, siphon it into a fermentation tank, add yeast and the soybean probiotic base liquid from step 1), and ferment at 20℃ for 6 days. 6) Soybean bleach pretreatment: Boil 8.7 L of fresh soybean bleach and then cool to room temperature; 7) Secondary fermentation: Add the soybean whey from step 6) to the fermentation liquid from step 5), add 150 g of sugar, and continue fermentation at 20℃ for 6 days; 8) Refrigeration: Place the fermented beer from step 7) in a refrigerator at 4°C for 24 hours to mature.

[0046] Comparative Example 6 (Pure Water Fermentation Comparative Example) A common probiotic beer has the following ingredients: 0.3 L soybean whey, 18 L purified water, 1.00 kg pale ale malt, 1.00 kg pale wheat malt, 200 g sucrose, 15 g Munich classic yeast, 3.00 g Lactobacillus plantarum 002, 10.00 g hops, and 2.00 g neutral protease. The preparation steps are as follows: 1) Preparation of soybean soy protein probiotic base solution: Boil 300 mL of fresh soybean soy protein and cool it to 30-40℃. Add 2.00 g of neutral protease and hydrolyze for 6 h. Then add 10 g of white sugar and sterilize at 105℃ for 8 min. Cool to 20℃ and then add Lactobacillus plantarum 002 bacterial solution. Incubate at 37℃ for 12 h and then store at 4℃ for later use. 2) Pure water treatment: Heat 9 L of pure water to 72-92℃; 3) Soaking and saccharification: Add the pure water from step 2) to the ground barley and wheat malt, and saccharify at 72°C for 1 hour to extract the wort; 4) Boil the wort: Boil the wort from step 3) for 60 minutes; 1st hop addition time: at the start of boiling, add 6.00 g of hops; 2nd hop addition time: after boiling for 30 minutes, add 4.00 g of hops; Hop variety: Magnum. 5) Primary fermentation: Let the wort from step 4) settle by vortexing for 15 minutes, cool to room temperature, siphon it into a fermentation tank, add yeast for fermentation, and ferment at 20℃ for 6 days; 6) Pretreatment: Boil 9 L of pure water and then cool it to room temperature; 7) Secondary fermentation: Add the pure water from step 6) to the fermentation liquid from step 5), add 190 g of sugar, add the soybean probiotic base liquid from step 1), and continue fermentation at 20℃ for 6 days; 8) Refrigeration: Place the fermented beer from step 7) at 4°C for 24 hours.

[0047] Comparative Example 7 (The formula is the same as Example 5, except that the steps for adding soybean bleach are different) A highly active probiotic soybean whey beer has the following ingredients: 10 L soybean whey, 10 L purified water, 1.00 kg pale ale malt, 1.00 kg pale wheat malt, 200 g sucrose, 10 g Munich classic yeast, 0.83 g Lactobacillus plantarum 002, 0.83 g Lactobacillus plantarum 376, 0.83 g STN Streptococcus thermophilus, 7.00 g hops, and 2.00 g neutral protease. The preparation steps are as follows: 1) Preparation of soybean probiotic base liquid: Same as in Example 5; 2) Soybean white water treatment: Collect 9.7 L of fresh soybean white water, add 10 L of pure water to make soybean white water solution, and then heat to 78℃; 3) Soaking and saccharification: Add the aqueous solution from step 2) to the ground wheat and barley malt, saccharify at 72°C for 1 hour, and then extract the wort; 4)-5) are the same as in Example 5; 6) Secondary fermentation: Weigh 190 g of sugar, add the soybean probiotic base liquid from step 1), and supplement the fermentation liquid from step 5). Continue fermentation at 20℃ for 6 days. 7) Refrigeration: Place the fermented beer from step 6) at 4°C for 24 hours.

[0048] Experimental Example: Performance testing of the products in each embodiment. 1. Sensory characteristics score of soy beer Table 1 Sensory Evaluation Grades of Bean Draft Beer (Unit: points)

[0049] Ten sensory evaluators were organized to evaluate the beer in terms of color, transparency, bitterness, carbonation, and acidity. As shown in Table 2, the color, transparency, ester aroma, hop aroma, bitterness, carbonation, aftertaste, purity, smoothness, mouthfeel, refreshingness, and acidity of the beer samples in each example were systematically evaluated.

[0050] Table 2 Sensory rating table for different bean beer

[0051] While the samples showed some differences in performance across various indicators, their overall flavor met the sensory quality requirements for beer. Example 1 exhibited superior ester aroma, a rich mouthfeel, pronounced fermentation flavor, and low acidity, making it suitable for consumers who prefer a milder, fruity taste. Comparative Example 6 had the strongest bitterness, as the use of water instead of whey resulted in a stronger bitterness and harshness.

[0052] 2. Colorimetric determination of soy beer The testing standard refers to GB / T 4928-2008, "Analytical Methods for Beer". Using a spectrophotometer, beer samples were injected into 10mm glass cuvettes, with water as a blank adjustment point. Wavelengths of 430nm and 700nm were measured respectively, and A... 430 ×0.039>A 700 This indicates that the beer is transparent. The absorbance value at 430nm was calculated, and the results are shown in Table 3.

[0053] Table 3 Colorimetric Analysis of Different Bean Flour Beers

[0054] 3. Turbidity determination of soybean beer The testing standard was based on GB / T 4928-2008, "Analytical Methods for Beer". The turbidity of the beer samples was measured after calibrating the turbidimeter with formalin standard turbidity solution, and the results are shown in Table 4.

[0055] Table 4. Turbidity Analysis of Different Bean Draft Beers

[0056] 4. Clarity of Bean Soft Beer The testing standard was based on GB / T4928-2008, the method for analyzing beer, and the results are shown in Table 5.

[0057] Table 5. Transparency Analysis of Different Bean Draft Beers

[0058] 5. Aroma determination of bean curd beer The testing standard was based on GB / T4928-2008, the method for analyzing beer, and the results are shown in Table 6.

[0059] Table 6 Aroma Analysis of Different Bean Stroke Beers

[0060]

[0061] As shown in Tables 3-6, the color values ​​ranged from a minimum of 26.50 (Example 4) to a maximum of 86.04 (Example 5); the turbidity ranged from a minimum of 22.70 (Comparative Example 3) to a maximum of 50.05 (Comparative Example 4); the transparency ranged from a darker, opaque beer (Example 2) to a pale yellow beer with higher transparency (Comparative Example 3); and the aroma ranged from a distinct beer aroma (Examples 3-5) to a lighter beer aroma. Significant differences in color, turbidity, transparency, and aroma were observed between the samples from different examples. These variations may be attributed to differences in brewing raw materials and processes, such as the specificity of the fermentation strains, the preparation of the soybean whey probiotic base, and the method of adding soybean whey. Example 1 exhibited significant differences in sensory evaluation due to the specific use of *Lactobacillus plantarum* 002 and its probiotic base, as well as the addition of soybean whey during the second fermentation.

[0062] 6. Electronic tongue measurement experiment 1) Sample pretreatment: The sample for electronic tongue testing must be a clear liquid and must not contain organic reagents; 2) Sensor preparation: taste sensor (S), reference solution (30 mmol / L potassium chloride + 0.3 mmol / L tartaric acid), internal solution (3.33 mol / L potassium chloride saturated silver chloride); 3) Sensor activation: The sensor was immersed in the solution for 24 h. The electrodes were soaked in a 3.3 mol / L potassium chloride solution, and the taste sensor was soaked in a reference solution for a slow soak. 4) Assemble the electronic tongue, install the taste sensor, and place the sample and reference solution; 5) Verify the sensor for 30 minutes before measurement; if the verification is successful, begin measuring the sample.

[0063] The electronic tongue detection results show (see) Figure 1 Comparative Examples 1 (D1) to 7 (D7) and Example 1 (S1) The beer prepared in Example 5 (S5) showed significant differences in sourness, astringency, umami, and saltiness. Among them, the sourness values ​​of Example 1 (S1), Comparative Example 6 (D6), Example 4 (S4), and Example 5 (S5) were lower. Meanwhile, the beers prepared in Comparative Example 6 (D6) and Example 4 (S4) had stronger bitterness, prominent umami, weak astringency, and no obvious aftertaste, with a balanced and distinct overall flavor.

[0064] 7. Electronic nose measurement experiment Experimental conditions: sampling time interval was 1 s / group, automatic sensor cleaning time was 120 s, sensor zeroing time was 5 s, injection flow rate was 600 mL / min, experimental test and analysis time was 60 s, and 28 different types of electronic nose sensors are shown in Table 7.

[0065] Table 7 Classification of Electronic Nose Sensors

[0066]

[0067] Electronic nose radar image analysis of the sample (see) Figure 2 (Table 7) Among the electronic nose receivers, sensors 1, 2, 20, and 26 are used for beer, respectively identifying alkanes, smoke, alcohols, aldehydes, short-chain alkanes, volatile organic compounds, alcohols, other small molecules, and organic acid derivatives such as esters and terpenes. Example 1 (S1) showed significantly higher response intensity than other samples on key sensors such as Sensor2 (alcohols, aldehydes, and short-chain alkanes), Sensor5 (nitrides and ammonia), Sensor20 (volatile organic compounds and alcohols), and Sensor26 (organic acid esters and terpenes), indicating that its volatile flavor compounds are richer and more complex.

[0068] 8. GC-MS detection of the flavor of bean beer Chromatographic conditions: InertCap column (30 m × 0.25 μm × 0.25 mm) was used with helium as carrier gas, injection port temperature was 250℃, injection time was 5 min, and splitless injection was used.

[0069] Sample preparation: Prepare a 2 g / L solution of 4-methyl-2-pentanol standard with anhydrous ethanol, and use it as an internal standard. Before testing, centrifuge the beer sample at low temperature to remove yeast from the beer to prevent interference with the test. Add 10 mL of beer sample to a centrifuge tube and centrifuge for 4 min (8000 rpm, 4℃). Add 3 g of sodium chloride to the headspace vial, then add 6 mL of the centrifuged beer sample. Add 20 μL of the internal standard to the headspace vial and cap it.

[0070] Detection: The sample was first equilibrated at 90°C for 20 min, followed by headspace extraction at 60°C for 30 min. The column temperature was initially 40°C, increased to 230°C at 2.5°C / min, and then to 240°C at 5°C / min. He was used as the carrier gas at a flow rate of 1 mL / min, splitless. The fiber was desorbed at 250°C in split-free mode for 3 min. MS detection was performed at 150°C in electron impact mode at 70 eV, with no solvent delay, and the ion source and transfer line temperatures were both 230°C.

[0071] The results, as shown in Table 8, indicate that the volatile components in beer include alcohols, esters, acids, ketones, aldehydes, and alkanes. The main volatile compounds are acids and esters, followed by ketones and alcohols, with small amounts of aldehydes and alkanes.

[0072] In Example 1, the contents of geraniol, ethyl acetate, and phenylethyl acetate were relatively high, while the contents of caprylic acid and beany aldehydes were relatively low. This is consistent with the results of Example 1 in the electronic tongue radar diagram. Geraniol is mainly produced by the transformation of terpenoids in hops and yeast metabolism, and has a fresh rose and citrus aroma. Phenylethyl acetate is an esterification product of phenylethanol and acetic acid during yeast fermentation, and has a sweet rose and honey aroma. Together, they make the beer aroma rich and layered. The above shows that the brewing process used in Example 1 and the interaction between Lactobacillus plantarum 002 and yeast increased the variety and quantity of volatile compounds in beer, resulting in better flavor.

[0073] Table 8. Analysis of the main volatile components of different bean beer

[0074]

[0075] 9. Nutritional composition analysis 9.1 Viable Bacterial Count Determination The viable counts of lactic acid bacteria, including *Lactobacillus plantarum*, *Streptococcus thermophilus*, and yeast in beer were determined using the national standard GB 4789.35-2023, Food Microbiology. The results are shown in Table 9.

[0076] Table 9. Analysis of viable probiotic counts in different soy beer varieties.

[0077]

[0078] After incubation at 37°C for 72 h, the total viable count of lactic acid bacteria and yeast in the beer of Example 1 was significantly higher than that in other examples and the comparative example, and the total viable count of Examples 2-5 was higher than 1×10⁻⁶. 7The CFU / mL count was highest in the beer from Example 1, which may be due to the specificity of *Lactobacillus plantarum* 002 as a lactic acid bacteria strain. Another possibility is that *Lactobacillus plantarum* 002 underwent initial adaptation and growth in soybean whey probiotic substrate prepared during the secondary fermentation process. The addition of soybean whey and probiotic substrate during this secondary fermentation allowed *Lactobacillus plantarum* 002 to thrive in the nutrient-rich environment of soybean whey, maintaining its probiotic activity. Furthermore, nutrient loss during the later stages of primary fermentation slowed the metabolism and activity of yeast, reducing the competitive inhibition of *Lactobacillus plantarum* 002 by yeast. This symbiotic relationship between the two promoted the maintenance of high bacterial activity in *Lactobacillus plantarum* 002.

[0079] 9.2 Detection of Raffinose and Stachyose Content The contents of raffinose and stachyose were determined by high performance liquid chromatography (HPLC), following the national standard GB / T 22491-2008 for soybean oligosaccharides. The detection results for each example are shown in Table 10. Figure 3 As shown.

[0080] Table 10 Analysis of Raffinose and Stachyose Content in Different Bean Stroke Beers

[0081]

[0082] Example 1 showed the highest content of raffinose and stachyose, with a significant increase compared to other examples. Raffinose and stachyose are excellent carbon sources for promoting the growth and reproduction of lactic acid bacteria. Therefore, the preparation of the Lactobacillus plantarum 002 soy-based culture medium and the fermentation specificity and secondary fermentation process in Example 1 not only promoted the reproduction and metabolism of Lactobacillus plantarum 002, degrading the natural polysaccharides in the raw materials into raffinose, but also promoted the growth of the strain and maintained high bacterial activity.

[0083] 9.3 Amino acid content detection The amino acid content in beer was determined by high performance liquid chromatography (HPLC), following the national standard GB 5009.124-2016, "Determination of Amino Acids in Food". The results are shown in Table 11.

[0084] Table 11. Analysis of Amino Acid Content in Different Bean Draft Beers (mg / L)

[0085]

[0086] Example 1 showed the highest total amino acid content. Furthermore, Example 1 had the highest leucine (Leu) content at 112.38 ± 0.86 mg / L, while Examples 2, Comparative Examples 1, 2, 6, and 7 had relatively lower leucine (Leu) contents. Leucine is an important prebiotic that shows a positive correlation with microbial proliferation capacity. As a branched-chain amino acid, leucine can promote the growth of lactic acid bacteria by regulating energy metabolism.

[0087] 9.4 Detection of Pyruvic Acid Content The pyruvate (PA) content was determined using the Solarbio reagent kit. 1 mL of extraction buffer was added to 0.5 mL of sample, and the sample was ultrasonically broken up while remaining in an ice bath. After ultrasonication, the sample was allowed to stand for 30 min, followed by centrifugation at 8000 g at room temperature for 10 min. The supernatant was then analyzed by UV-Vis at 520 nm to calculate the pyruvate content. The results are shown in Table 12.

[0088] Table 12 Analysis of Pyruvic Acid Content in Different Soft Beers

[0089] Example 1 showed the highest pyruvate content. Pyruvate is not only the end product of glycolysis but also plays a crucial role in the synthesis and breakdown of amino acids. In Example 1, the active probiotic flora exhibited enhanced metabolic capacity, promoting energy metabolism and amino acid synthesis. Pyruvate may accept the amino group of glutamate through alanine aminotransferase-mediated transamination to produce α-ketoglutarate. This reaction provides an important amino skeleton for amino acid synthesis. Furthermore, as a three-carbon α-keto acid skeleton, pyruvate is an important carbon source for branched-chain amino acids such as leucine, participating in their synthesis pathways through multiple enzymatic reactions, thereby enriching the variety and content of amino acids in the sample. Amino acids not only provide the necessary nitrogen source for the growth of lactic acid bacteria but also serve as substrates for their cellular metabolism, supporting the growth and activity of lactic acid bacteria.

[0090] 10. Physicochemical index analysis 10.1, Alcohol content The testing standard was based on GB / T 4928-2008, the analytical method for beer. The results are shown in Table 13.

[0091] Table 13 Analysis of alcohol content of different bean beer

[0092] The alcohol content of beer is generally positively correlated with the wort concentration; the higher the wort concentration, the higher the alcohol content, and the richer the nutritional value of the beer. The detected alcohol content is consistent with the changes in wort concentration.

[0093] 10.2 Original wort concentration The testing standard followed GB / T 4928-2008, the analytical method for beer. The results are shown in Table 14.

[0094] Table 14 Analysis of Original Wort Concentration in Different Bean Stroke Beers

[0095] Example 1 had a higher original wort concentration. Beer color is related to wort concentration; a higher wort concentration indicates better nutritional value and richer flavor, further demonstrating more thorough probiotic fermentation. A higher degree of beer fermentation means that the yeast converted more sugar into alcohol during fermentation.

[0096] 10.3 Total acid The testing standard followed GB / T 4928-2008, the analytical method for beer. The results are shown in Table 15.

[0097] Table 15 Analysis of Total Acidity of Different Bean Draft Beers

[0098] The total acid values ​​of Examples 1 and Comparative Example 6 were relatively low and mild. The total acid values ​​of Examples 2-5 were moderately high, which is consistent with the trend of acid content in the electronic tongue acid intensity and gas chromatography results.

[0099] 10.4 True degree of fermentation and invertase activity The testing standard was based on GB / T 4928-2008, the analytical method for beer. The results are shown in Table 16.

[0100] Table 16 Analysis of True Fermentation Degree and Invertase Activity in Different Bean Stroke Beers

[0101]

[0102] The actual degree of fermentation was relatively low in all embodiments, which may be due to the symbiotic relationship between yeast and lactic acid bacteria during the brewing process of probiotic beer. The slow growth of yeast interacts with the high activity of lactic acid bacteria, which may lead to incomplete sugar conversion, thereby promoting a lower degree of fermentation and giving the beer its unique flavor and nutritional characteristics. In addition, the invertase activity was positive in all embodiments. This may be because various enzyme systems remain viable in unpasteurized beer, and the identification of invertase activity proves that the beer is a craft beer.

[0103] Although preferred embodiments of the invention have been described, those skilled in the art, upon learning the basic inventive concept, can make other changes and modifications to these embodiments. Therefore, the appended claims are intended to be interpreted as including both the preferred embodiments and all changes and modifications falling within the scope of the invention.

[0104] Obviously, those skilled in the art can make various modifications and variations to this invention without departing from its spirit and scope. Therefore, if these modifications and variations fall within the scope of the claims of this invention and their equivalents, this invention also intends to include these modifications and variations.

Claims

1. A highly active probiotic soybean whey beer, characterized in that, The following components are included in parts by weight: 8000-10000 parts soybean whey, 8000-10000 parts pure water, 1000-2000 parts barley malt, 800-1000 parts wheat malt, 100-200 parts sugar, 6-10 parts hops, 1-2 parts neutral protease, 5-15 parts yeast, and 1-3 parts lactic acid bacteria.

2. The highly active probiotic soybean whey beer according to claim 1, characterized in that, The yeast strain is Munich brewer's yeast; The hops mentioned are Magnum hops.

3. The highly active probiotic soybean whey beer according to claim 1, characterized in that, The soybean whey is obtained by separating and separating soybean whey after the soybean protein has coagulated or precipitated during the processing of soybean products using salt, acid and microbial fermentation.

4. The highly active probiotic soybean whey beer according to claim 1, characterized in that, The lactic acid bacteria are one or more of Lactobacillus plantarum and Streptococcus thermophilus.

5. The highly active probiotic soybean whey beer according to claim 1, characterized in that, The highly active probiotic soybean whey beer contains 1×10⁻⁶ live probiotics. 7 CFU / mL.

6. A method for preparing highly active probiotic soybean whey beer according to any one of claims 1-5, characterized in that, The specific steps include the following: Preparation of soybean probiotic base liquid: Boil 300-400 parts of soybean protein and cool to 30-40℃ to obtain soybean protein base liquid; After the soybean protein base solution was hydrolyzed with neutral protease for 4-6 hours, 10 parts of sugar were added, and the solution was sterilized at 105℃ and then cooled to obtain soybean protein culture medium. Add the bacterial solution containing lactic acid bacteria to the soybean culture medium and incubate at 37°C for 8-12 hours to prepare soybean probiotic base solution; Soaking and saccharification: Add heated pure water to the mixture of ground barley malt and wheat malt, and saccharify at 72-82℃ for 1 hour to obtain wort; Boiling the wort: Boil the wort. At the beginning of boiling, add hops at a mass ratio of 3 / 6 to 4 / 6. After boiling for 30 to 40 minutes, add the remaining hops and boil for another 30 to 40 minutes to obtain hop wort. Primary fermentation: Cool the hop wort to room temperature, add yeast and ferment at 20°C for 6-8 days to obtain primary fermentation liquid; Secondary fermentation: After boiling and cooling the remaining soybean whey, mix it with the soybean whey probiotic base liquid, add the remaining sugar, stir evenly to obtain soybean whey fermentation liquid, add the soybean whey fermentation liquid to the primary fermentation liquid, and continue fermentation at 20℃ for 6-8 days to obtain the highly active probiotic soybean whey beer. Refrigeration: The highly active probiotic soybean beer is refrigerated at 4°C for post-maturation.

7. The method for preparing a highly active probiotic soybean whey beer according to claim 6, characterized in that, The preparation of the lactic acid bacteria culture includes: inoculating lactic acid bacteria powder into MRS broth and culturing at 37°C for 24 h; then preparing a second subculture in the same manner to obtain the culture, with a viable count > 1 × 10⁻⁶. 7 CFU / mL.

8. A method for preparing a highly active probiotic soybean whey beer according to claim 6, characterized in that, The highly active probiotic soybean beer was aged for 24 hours after refrigeration.

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