Beer rich in radix cynanchi bungei and preparation method thereof

Abstract

The invention relates to the technical field of beer brewing, in particular to beer rich in radix cynanchi bungei and a preparation method thereof.The preparation method includes the steps that barley malt, wheat malt and radix cynanchi bungei are smashed, mixed, saccharified, filtered in wort, boiled, subjected to rotary precipitation, cooled, fermented, bottled and sterilized; the radix cynanchi bungei beer is good in non-biological stability, obvious in radix cynanchi bungei characteristic taste, free of bitter taste and good in oxidation resistance. The two kinds of humulus lupulus are added during boiling and fermentation respectively, basic bitter taste is provided for the beer, meanwhile, the special flavor of the radix cynanchi bungei is compatible with the overall flavor of the beer as much as possible, the obvious bitter taste of the radix cynanchi bungei beer is relieved, and the overall taste and the three-dimensional level are improved; the contained compound enzyme preparation can promote the saccharification process and fully release active substances in the radix cynanchi bungei, the non-biological stability of the beer can be improved by reducing the turbidity value, and compared with the prior art, the compound enzyme preparation has wide application prospects.

Description

Technical Field

[0001] The invention relates to the technical field of beer brewing, in particular to beer rich in Cynanchum multiflorum and a preparation method thereof. Background Art

[0002] Beer is a low-alcohol beverage saturated with carbon dioxide, brewed from malt, hops, and water through yeast fermentation. It is nutritious, refreshing, and has a unique flavor, often referred to as liquid bread. As one of the world's three most ancient alcoholic beverages, beer has become an integral part of human culture and social development. Its brewing techniques have evolved significantly through millennia of inheritance and innovation.

[0003] Specialty beers are brewed with unique styles by modifying raw materials, auxiliary materials, and production processes. Currently, these beers primarily rely on traditional brewing methods, leveraging new food resources rich in bioactive or flavorful substances such as flavonoids, polysaccharides, dietary fiber, volatile oils, and polyphenols. By modifying processes such as gelatinization, saccharification, and fermentation, specialty beers are brewed with flavors and nutritional properties distinct from those of traditional craft beers. Beer contains at least 17 common amino acids, particularly all essential amino acids. However, the types and amounts of these amino acids vary significantly depending on the brewing ingredients and yeast used.

[0004] Baishouwu, also known as ear-leafed cow-skin shavings, mountain-crossing shavings, and milk-skinned vine, is the tuberous root of the dicotyledonous plant Cyperus rotundus, part of the Apocynaceae family. It grows in mountain thickets, under forests on hillsides, along roadsides, in forest-edge meadows, and is also found in fields and wasteland. Baishouwu is a traditional Chinese medicinal and edible plant, and a local, authentic Chinese herbal medicine. It is rich in a variety of nutrients, including starch, polysaccharides, amino acids, and trace elements, as well as bioactive components such as C21 steroidal glycosides, flavonoids, and acetophenones. It exhibits anti-tumor, antioxidant, liver and kidney protection, lipid-lowering, and immune-regulating properties.

[0005] Currently, the majority of Baishouwu products are primarily processed slices and refined powder. These products suffer from issues such as poor ready-to-eat properties, a strong herbal flavor, and poor taste. The refined powder processing, in particular, results in significant loss of active ingredients. To address these issues with primarily processed Baishouwu products, the use of microbial fermentation technology to improve the flavor, nutritional value, and added value of Baishouwu has become a new trend in Baishouwu processing. New products, such as Baishouwu wine, fermented milk, and enzymes, are now being developed or launched.

[0006] In the prior art, for example, patent document CN113831972A discloses a high auxiliary material ratio Binhai Baishouwu beer and a preparation method thereof. In this invention, the starch-rich property of Baishouwu is utilized, and Baishouwu starch is used as the main saccharification raw material. Beer is brewed at a ratio of 1:1 between Baishouwu and barley malt, and external enzymes such as cellulase and pullulanase are selected to enhance the saccharification of Baishouwu starch. At the same time, the mixed saccharification liquid of Baishouwu and malt is co-fermented to obtain a Baishouwu beer rich in arginine. In this invention, a variety of external enzymes are used to saccharify Baishouwu, which can effectively reduce the influence of Baishouwu bitter substances on the overall flavor of beer. However, the ratio of Baishouwu to barley malt in the raw materials used in this invention is 1:1, and Baishouwu contains a large amount of polyphenols. While improving the overall antioxidant properties and Baishouwu flavor, it has a certain influence on the non-biological stability of beer.

[0007] Therefore, according to the related technology mentioned above, it is urgent to develop a beer rich in Cynanchum multiflorum and a preparation method thereof. Summary of the Invention

[0008] In view of this, the purpose of the present invention is to provide a beer rich in Cynanchum multiflorum and a preparation method thereof, so as to solve the problems of obvious bitterness and poor non-biological stability of Cynanchum multiflorum beer in the prior art.

[0009] Based on the above purpose, the present invention provides a beer rich in Cynanchum multiflorum and a preparation method thereof.

[0010] A beer rich in Cynanchum multiflorum comprises the following raw materials in parts by weight: 62-70 parts of barley malt; 50-57 parts of wheat malt; 30-40 parts of Cynanchum multiflorum; 0.32-0.42 parts of grape leaves; 0.8-1.3 parts of complex enzyme preparation; 8.2-16.5 parts of yeast composition; 0.1-0.14 parts of Cascade hops; 0.27-0.3 parts of Simcoe hops; The yeast composition is prepared from Pichia kluyveri and Wickham M3 yeast; The complex enzyme preparation comprises α-amylase, endoproline enzyme, saccharifying enzyme and β-glucanase in a mass ratio of 8-11.2:2.4-3:6.5-7.2:3.3-4; The activity of α-amylase in the complex enzyme preparation is 220-270 U / g; The activity of the proline endoenzyme in the complex enzyme preparation is 95-110 U / g; The activity of the saccharifying enzyme in the complex enzyme preparation is 15.5-17.2 U / g; The activity of the β-glucanase in the complex enzyme preparation is 6-7 U / g.

[0011] Preferably, the preparation method of the yeast composition is as follows: Step A1. Pichia kluyveri and Wickham's M3 yeast were inoculated into YPD solid medium, cultured at 30-35°C for 20-36 hours to obtain activated strains, and then inoculated into YPD liquid medium and cultured at 30-37°C and a shaker speed of 160-180 r / min for 20-36 hours to obtain a first-level seed solution of Pichia kluyveri and a first-level seed solution of Wickham's M3 yeast; Step A2. The first-level seed liquid of Pichia kluyveri and the first-level seed liquid of Wickham's M3 yeast were respectively inoculated into a fermentation tank containing YPD liquid medium for cultivation to obtain a second-level seed liquid of Pichia kluyveri and a second-level seed liquid of Wickham's M3 yeast. The second-level seed liquid of Pichia kluyveri and the second-level seed liquid of Wickham's M3 yeast were mixed evenly at an effective viable count of 2-3:0.5-1, and centrifuged at 4000-5000 rpm for 5-8 min, and then washed twice with 0.85% sterile NaCl solution to obtain a yeast composition.

[0012] Preferably, the number of viable Pichia kluyveri in the secondary seed solution of Pichia kluyveri in step A2 is 3.2-4.6×10 8 cfu / mL.

[0013] Preferably, the volume ratio of the Pichia kluyveri primary seed solution to the YPD liquid medium in step A2 is 100:700-900; The volume ratio of the abnormal Wickham M3 yeast first-level seed liquid to the YPD liquid culture medium is 100:700-900.

[0014] A method for preparing beer rich in Radix Cynanchum multiflorum comprises the following steps: Step S1. Washed radix polygoni multiflori is cut into thin slices with a thickness of 3-5 mm, pre-frozen at -20-15°C for 1-2 hours, freeze-dried, and then pulverized at 4°C to obtain radix polygoni multiflori powder with a fineness of 0.1-0.15 mm; barley malt and wheat malt are pulverized by a moisture recovery pulverization method to obtain wheat flour with a fineness of 0.15-0.2 mm, which is thoroughly mixed with the radix polygoni multiflori powder to obtain a mixture 1; Step S2. Add brewing water to the saccharification cup and add lactic acid to adjust the pH of the system to 4-5. Raise the system temperature to T1 and add Mix 1 while stirring. Keep warm for 30-40 minutes. Then, perform a first heating and keep warm for 50-60 minutes. Then, perform a second heating and keep warm for 25-30 minutes. Cool naturally to T2 to obtain Mix 2. Step S3. Add the complex enzyme preparation to the mixture 2, keep warm at temperature T2 and perform an iodine test. After the iodine test is normal, heat for the third time, keep warm for 15-20min, filter, and obtain wort; Step S4. The wort is boiled for 74-82 minutes, and Simcoe hops and grape leaves with a fineness of 0.24-0.28 mm and freeze-dried Cynanchum multiflorum powder with a fineness of 0.1-0.15 mm are added. After boiling, the wort is filtered using a conventional lauter tun, rinsed twice with the lauter tun water at a temperature of 75-78°C, and then vortexed for 10-15 minutes. After cooling, the wort concentration is adjusted to 11°P, and the wort is boiled and sterilized for 20-30 minutes and then cooled to room temperature to obtain clarified wort. Step S5. The yeast composition is added to the clarified wort, and the wort is placed in a fermenter for a primary fermentation stage. The wort is first fermented at 10-12°C for 4-5 days. The wort is sealed when the residual sugar drops to 4°C. After 2 days, the wort is cooled by 1°C every 3 hours until it drops to 2-4°C. A secondary fermentation is carried out for 7 days until the diacetyl content is ≤0.05 mg / L. The fermentation is terminated, and Cascade hops are dry-added. After soaking for 3-5 days, the beer is filtered, bottled, and pasteurized to obtain a beer rich in Radix Polygoni Multiflori.

[0015] Preferably, the freeze-drying in step S1 includes a first freezing stage and a second drying stage; The temperature during the first freezing stage is -42-35°C and the time is 5-6.5 hours; The temperature during the second drying stage is -15-0°C and the time is 15-18 hours.

[0016] Preferably, the temperature T1 in step S2 is 38-44°C; The heating rate during the first heating is 1-2°C / min, and the temperature is raised to 52-56°C; The second heating process is carried out at a heating rate of 1-2°C / min to a temperature of 90-94°C.

[0017] Preferably, the temperature T2 in step S2 is 62-65°C; The ratio of brewing water to mixed material 1 is 530-680 mL:140-163 g.

[0018] Preferably, the heating rate during the third heating in step S3 is 1-2°C / min, and the temperature is raised to 70-74°C.

[0019] Preferably, the Cynanchum multiflorum powder and grape leaves in step S4 are added at the beginning of boiling; The Simcoe hops were added in three times: one-half of the hops was added at 10 min of boiling for the first time, one-quarter of the hops was added at 40 min of boiling for the second time, and one-quarter of the hops was added at 65 min of boiling for the third time.

[0020] Beneficial effects of the present invention: The invention provides a beer rich in Radix Angelicae Multiflori and a preparation method thereof. In the invention, barley malt, wheat malt and Radix Angelicae Multiflori are crushed, mixed, saccharified, wort filtered, boiled, vortex precipitated, cooled, fermented, bottled and then sterilized to obtain Radix Angelicae Multiflori beer with good non-biological stability, obvious Radix Angelicae Multiflori characteristic taste, no bitterness and good antioxidant properties. Two types of hops are added during boiling and fermentation respectively, which not only provide basic bitterness for the beer but also maximize the compatibility of the special flavor of Radix Angelicae Multiflori with the overall flavor of the beer, reduce the obvious bitterness of the Radix Angelicae Multiflori beer, and enhance the overall taste and three-dimensional layers. In addition, the synergistic effect of α-amylase, proline endonuclease, saccharifying enzyme and β-glucanase in the complex enzyme preparation promotes saccharification and fully releases active substances in Radix Angelicae Multiflori. At the same time, the proline endonuclease improves the non-biological stability of the beer by reducing the turbidity value. Compared with the existing technology, the invention has broad application prospects. DETAILED DESCRIPTION

[0021] In order to make the objectives, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below with reference to specific embodiments.

[0022] The sources and properties of some of the raw materials used in the present invention are as follows: Endoproline enzyme was purchased from Qingdao Weilan Biotechnology Co., Ltd.; β-glucanase was purchased from Qingdao Weilan Biotechnology Co., Ltd.; Cascade 90 hops were purchased from Yakima Hop Company, USA; and Simcoe 90 hops were purchased from Yakima Hop Company, USA.

[0023] The abnormal Wickham yeast M3 used in the present invention is disclosed in the invention patent with the authorization publication number "CN117801969A" and the name "An abnormal Wickham yeast strain capable of increasing the aroma of wine brewing", and the deposit number is CGMCC No. 28666; The Pichia kluyveri used in the present invention is Pichia kluyveri JLG30201, which is disclosed in the invention patent with authorization publication number “CN119752652A” and titled “A Pichia kluyveri and its application”, and has a deposit number of CCTCC NO: M 20242295.

[0024] Example 1: A method for preparing beer rich in Radix Cynanchifolii, comprising the following steps: S1. Weigh 10 g of yeast extract, 20 g of peptone, and 20 g of glucose into distilled water, stir until completely dissolved, dilute to 1 L, adjust the pH to 6.5, add 10 g of agar, heat at 50°C until completely dissolved, filter, and autoclave at 121°C for 15 min to obtain YPD solid medium. Weigh 10 g of yeast extract, 20 g of peptone, and 20 g of glucose into distilled water, stir until completely dissolved, dilute to 1 L, adjust the pH to 6.5, filter, and autoclave at 121°C for 15 min to obtain YPD liquid medium. S2. Pichia kluyveri JLG30201 and Wickham's M3 yeast were inoculated into YPD solid medium and cultured at 30°C for 20 hours to obtain activated strains. The strains were then inoculated into YPD liquid medium and cultured at 30°C with a shaker at 160 rpm for 20 hours to obtain primary seed solutions of Pichia kluyveri and Wickham's M3 yeast. S3. 100 mL of a Pichia kluyveri primary seed solution and 100 mL of an abnormal Wickham M3 yeast primary seed solution were inoculated into a fermenter containing 700 mL of YPD liquid medium to obtain a secondary Pichia kluyveri seed solution and an abnormal Wickham M3 yeast secondary seed solution. The secondary Pichia kluyveri seed solution and the abnormal Wickham M3 yeast secondary seed solution were mixed at an effective viable count of 2:0.5 and centrifuged at 4000 rpm for 5 min, and then washed twice with 0.85% sterile NaCl solution to obtain a yeast composition. S4. Take the washed Cynanchum multiflorum and cut it into slices with a thickness of 3 mm. After pre-freezing at -20 ° C for 1 h, freeze-drying was performed. The temperature of the first freezing stage was -42 ° C for 5 h; the temperature of the second drying stage was -15 ° C for 15 h; then the mixture was crushed at 4 ° C to obtain a Cynanchum multiflorum powder with a fineness of 0.1 mm; 62 g of barley malt and 50 g of wheat malt were crushed by moisture regeneration method to obtain wheat flour with a fineness of 0.15 mm, which was thoroughly mixed with 28 g of Cynanchum multiflorum powder to obtain a mixture 1; S5. Add 530 mL of brewing water to the saccharification cup, and add lactic acid to adjust the pH of the system to 4. Raise the system temperature to 38°C, add 140 g of Mix 1 while stirring, and keep warm for 30 minutes. Then, perform a first heating at a rate of 1°C / min to 52°C and keep warm for 50 minutes. Then perform a second heating at a rate of 1°C / min to 90°C, keep warm for 25 minutes, and cool naturally to 62°C to obtain Mix 2. S6. 8 g of α-amylase with an activity of 220 U / g, 2.4 g of proline endoenzyme with an activity of 95 U / g, 6.5 g of saccharifying enzyme with an activity of 15.5 U / g, and 3.3 g of β-glucanase with an activity of 6 U / g were mixed to obtain a complex enzyme preparation; 0.8 g of the complex enzyme preparation was added to the mixture 2, and the mixture was kept warm at 62 ° C and iodine test was performed. After the iodine test was normal, the temperature was raised for the third time at a heating rate of 1 ° C / min, and the temperature was raised to 70 ° C, kept warm for 15 min, and filtered to obtain wort; S7. The wort was boiled for 74 min and 0.27 g of Simcoe hops and 0.32 g of grape leaves with a fineness of 0.24 mm were added, as well as 2 g of freeze-dried Cynanchum multiflorum powder with a fineness of 0.1 mm, wherein the Cynanchum multiflorum powder and grape leaves were added at the start of boiling, and Simcoe hops were added in three portions, the first time half was added at 10 min, the second time a quarter was added at 40 min, and the third time a quarter was added at 65 min; after boiling, the wort was filtered using a conventional filter tank, the tank was washed twice and the water temperature of the wash tank was 75 ° C, and then vortexed for 10 min, and the wort concentration was adjusted to 11 ° P after cooling, and the wort was sterilized by boiling for 20 min and then cooled to room temperature to obtain clarified wort; S8. 8.2 g of the yeast composition is added to the clarified wort, and the wort is placed in a fermenter for a primary fermentation stage. The wort is first fermented at 10°C for 4 days. The fermentation tank is sealed when the residual sugar drops to 4°P. After 2 days, the temperature is lowered by 1°C every 3 hours until it drops to 2°C. A secondary fermentation is carried out for 7 days until the diacetyl content is ≤0.05 mg / L. The fermentation is terminated, and 0.1 g of Cascade hops is dry-doped into the wort. After soaking for 3 days, the beer is filtered, bottled, and pasteurized to obtain a beer rich in Radix Polygoni Multiflori.

[0025] Example 2: A method for preparing beer rich in Radix Cynanchifolii, comprising the following steps: S1. Weigh 10 g of yeast extract, 20 g of peptone, and 20 g of glucose into distilled water, stir until completely dissolved, dilute to 1 L, adjust the pH to 6.5, add 10 g of agar, heat at 50°C until completely dissolved, filter, and autoclave at 121°C for 15 min to obtain YPD solid medium. Weigh 10 g of yeast extract, 20 g of peptone, and 20 g of glucose into distilled water, stir until completely dissolved, dilute to 1 L, adjust the pH to 6.5, filter, and autoclave at 121°C for 15 min to obtain YPD liquid medium. S2. Pichia kluyveri JLG30201 and Wickham's M3 yeast were inoculated into YPD solid medium and cultured at 32°C for 25 hours to obtain activated strains. These strains were then inoculated into YPD liquid medium and cultured at 32°C with a shaker at 165 rpm for 25 hours to obtain primary seed solutions of Pichia kluyveri and Wickham's M3 yeast. S3. 100 mL of a Pichia kluyveri primary seed solution and 100 mL of an abnormal Wickham M3 yeast primary seed solution were inoculated into a fermenter containing 800 mL of YPD liquid medium to obtain a secondary Pichia kluyveri seed solution and an abnormal Wickham M3 yeast secondary seed solution. The secondary Pichia kluyveri seed solution and the abnormal Wickham M3 yeast secondary seed solution were mixed at an effective viable count of 2:1 and centrifuged at 4500 rpm for 6 min, followed by washing twice with 0.85% sterile NaCl solution to obtain a yeast composition. S4. Take the washed Cynanchum multiflorum and cut it into slices with a thickness of 4 mm. After pre-freezing at -15 ° C for 1 h, freeze-drying was performed. The temperature of the first freezing stage was -38 ° C for 5.5 h; the temperature of the second drying stage was -5 ° C for 16 h; then the mixture was pulverized at 4 ° C to obtain a Cynanchum multiflorum powder with a fineness of 0.12 mm; 64 g of barley malt and 52 g of wheat malt were pulverized by moisture regeneration to obtain wheat flour with a fineness of 0.17 mm, which was thoroughly mixed with 31 g of Cynanchum multiflorum powder to obtain a mixture 1; S5. Add 580 mL of brewing water to the saccharification cup, and add lactic acid to adjust the pH of the system to 4. Raise the system temperature to 40°C, add 147 g of Mix 1 while stirring, and keep warm for 33 minutes. Then, perform a first heating at a rate of 1°C / min to 54°C and keep warm for 53 minutes. Then perform a second heating at a rate of 1°C / min to 92°C, keep warm for 27 minutes, and cool naturally to 64°C to obtain Mix 2. S6. 9 g of α-amylase with an activity of 240 U / g, 2.6 g of proline endoenzyme with an activity of 100 U / g, 6.8 g of saccharifying enzyme with an activity of 16.2 U / g, and 3.5 g of β-glucanase with an activity of 6.3 U / g were mixed to obtain a complex enzyme preparation; 1 g of the complex enzyme preparation was added to the mixture 2, and the mixture was kept warm at 63 ° C and subjected to an iodine test. After the iodine test was normal, the temperature was raised for the third time at a heating rate of 1 ° C / min to 72 ° C, kept warm for 18 min, and filtered to obtain wort; S7. The wort was boiled for 76 min and 0.28 g of Simcoe hops and 0.35 g of grape leaves with a fineness of 0.26 mm were added, as well as 2.7 g of freeze-dried Cynanchum multiflorum powder with a fineness of 0.12 mm, wherein the Cynanchum multiflorum powder and grape leaves were added at the start of boiling, and Simcoe hops were added in three portions, the first time half was added at 10 min, the second time a quarter was added at 40 min, and the third time a quarter was added at 65 min; after boiling, the wort was filtered using a conventional lauter tun, the trough was washed twice and the temperature of the wash water was 76 ° C, and then vortexed and precipitated for 12 min, and the wort concentration was adjusted to 11 ° P after cooling, and the wort was sterilized by boiling for 23 min and then cooled to room temperature to obtain clarified wort; S8. 11.5 g of the yeast composition is added to the clarified wort, and the wort is placed in a fermenter for the main fermentation stage. The wort is first fermented at 10°C for 4 days. The tank is sealed when the residual sugar drops to 4°P. After 2 days, the temperature is lowered by 1°C every 3 hours until it drops to 3°C. A post-fermentation is carried out for 7 days until the diacetyl content is ≤0.05 mg / L. The fermentation is terminated, and 0.11 g of Cascade hops is dry-doped into the wort. After soaking for 4 days, the beer is filtered, bottled, and pasteurized to obtain a beer rich in Radix Polygoni Multiflori.

[0026] Example 3: A method for preparing beer rich in Radix Cynanchifolii, comprising the following steps: S1. Weigh 10 g of yeast extract, 20 g of peptone, and 20 g of glucose into distilled water, stir until completely dissolved, dilute to 1 L, adjust the pH to 6.5, add 10 g of agar, heat at 50°C until completely dissolved, filter, and autoclave at 121°C for 15 min to obtain YPD solid medium. Weigh 10 g of yeast extract, 20 g of peptone, and 20 g of glucose into distilled water, stir until completely dissolved, dilute to 1 L, adjust the pH to 6.5, filter, and autoclave at 121°C for 15 min to obtain YPD liquid medium. S2. Pichia kluyveri JLG30201 and Wickham's M3 yeast were inoculated into YPD solid medium and cultured at 32°C for 30 hours to obtain activated strains. These strains were then inoculated into YPD liquid medium and cultured at 35°C with a shaker at 170 rpm for 30 hours to obtain primary seed solutions of Pichia kluyveri and Wickham's M3 yeast. S3. 100 mL of a Pichia kluyveri primary seed solution and 100 mL of an abnormal Wickham M3 yeast primary seed solution were inoculated into a fermenter containing 800 mL of YPD liquid medium to obtain a secondary Pichia kluyveri seed solution and an abnormal Wickham M3 yeast secondary seed solution. The secondary Pichia kluyveri seed solution and the abnormal Wickham M3 yeast secondary seed solution were mixed at an effective viable count of 3:0.5 and centrifuged at 4500 rpm for 7 min, followed by washing twice with 0.85% sterile NaCl solution to obtain a yeast composition. S4. Take the washed Cynanchum multiflorum and cut it into slices with a thickness of 4 mm. After pre-freezing at -18 ° C for 2h, freeze-drying was performed. The temperature of the first freezing stage was -40 ° C for 6h; the temperature of the second drying stage was -10 ° C for 17h; then the mixture was crushed at 4 ° C to obtain a Cynanchum multiflorum powder with a fineness of 0.14mm; 67g of barley malt and 55g of wheat malt were crushed by moisture regeneration method to obtain wheat flour with a fineness of 0.18mm, which was thoroughly mixed with 33g of Cynanchum multiflorum powder to obtain a mixture 1; S5. Add 630 mL of brewing water to the saccharification cup, add lactic acid to adjust the pH of the system to 5, raise the system temperature to 42°C, add 155 g of Mix 1 while stirring, and keep warm for 37 minutes. Then, perform a first heating at a rate of 2°C / min to 55°C and keep warm for 57 minutes. Then perform a second heating at a rate of 2°C / min to 94°C and keep warm for 30 minutes. Then, cool naturally to 65°C to obtain Mix 2. S6. 10.1 g of α-amylase with an activity of 260 U / g, 2.8 g of proline endoenzyme with an activity of 105 U / g, 7 g of saccharifying enzyme with an activity of 16.7 U / g, and 3.8 g of β-glucanase with an activity of 6.7 U / g were mixed to obtain a complex enzyme preparation; 1.1 g of the complex enzyme preparation was added to the mixture 2, and the mixture was kept warm at 65 ° C and iodine test was performed. After the iodine test was normal, the temperature was raised for the third time at a heating rate of 2 ° C / min, and the temperature was raised to 74 ° C, kept warm for 20 min, and filtered to obtain wort; S7. The wort was boiled for 89 min and 0.29 g of Simcoe hops and 0.39 g of grape leaves with a fineness of 0.26 mm were added, as well as 3.5 g of freeze-dried Cynanchum multiflorum powder with a fineness of 0.14 mm, wherein the Cynanchum multiflorum powder and grape leaves were added at the start of boiling, and Simcoe hops were added three times, the first time half was added at 10 min, the second time a quarter was added at 40 min, and the third time a quarter was added at 65 min; after boiling, the wort was filtered using a conventional filter tank, the tank was washed twice and the temperature of the wash water was 78 ° C, and then vortexed for 15 min, and the wort concentration was adjusted to 11 ° P after cooling, and the wort was sterilized by boiling for 27 min and then cooled to room temperature to obtain clarified wort; S8. 13.5 g of the yeast composition is added to the clarified wort, and the wort is placed in a fermenter for the main fermentation stage. The wort is first fermented at 12° C. for 5 days. The fermentation tank is sealed when the residual sugar drops to 4° C. After 2 days, the temperature is lowered by 1° C. every 3 hours until it reaches 4° C. A post-fermentation is carried out for 7 days until the diacetyl content is ≤0.05 mg / L. The fermentation is terminated, and 0.14 g of Cascade hops is dry-doped into the wort. After soaking for 5 days, the beer is filtered, bottled, and pasteurized to obtain a beer rich in Radix Polygoni Multiflori.

[0027] Example 4: A method for preparing beer rich in Radix Cynanchifolii, comprising the following steps: S1. Weigh 10 g of yeast extract, 20 g of peptone, and 20 g of glucose into distilled water, stir until completely dissolved, dilute to 1 L, adjust the pH to 6.5, add 10 g of agar, heat at 50°C until completely dissolved, filter, and autoclave at 121°C for 15 min to obtain YPD solid medium. Weigh 10 g of yeast extract, 20 g of peptone, and 20 g of glucose into distilled water, stir until completely dissolved, dilute to 1 L, adjust the pH to 6.5, filter, and autoclave at 121°C for 15 min to obtain YPD liquid medium. S2. Pichia kluyveri JLG30201 and Wickham's M3 yeast were inoculated into YPD solid medium and cultured at 35°C for 36 hours to obtain activated strains. These strains were then inoculated into YPD liquid medium and cultured at 37°C with a shaker at 180 rpm for 36 hours to obtain primary seed solutions of Pichia kluyveri and Wickham's M3 yeast. S3. 100 mL of a Pichia kluyveri primary seed solution and 100 mL of an abnormal Wickham M3 yeast primary seed solution were inoculated into a fermenter containing 900 mL of YPD liquid medium to obtain a secondary Pichia kluyveri seed solution and an abnormal Wickham M3 yeast secondary seed solution. The secondary Pichia kluyveri seed solution and the abnormal Wickham M3 yeast secondary seed solution were mixed at an effective viable count of 3:1 and centrifuged at 5000 rpm for 8 min, followed by washing twice with 0.85% sterile NaCl solution to obtain a yeast composition. S4. Take the washed Cynanchum multiflorum and cut it into slices with a thickness of 5 mm. After pre-freezing at -20 ° C for 2h, freeze-drying was performed. The temperature of the first freezing stage was -42 ° C for 6.5h; the temperature of the second drying stage was -15 ° C for 18h; then the mixture was crushed at 4 ° C to obtain a Cynanchum multiflorum powder with a fineness of 0.15mm; 70g of barley malt and 57g of wheat malt were crushed by moisture regeneration method to obtain wheat flour with a fineness of 0.2mm, which was thoroughly mixed with 36g of Cynanchum multiflorum powder to obtain a mixture 1; S5. Add 680 mL of brewing water to the saccharification cup, add lactic acid to adjust the pH of the system to 5, raise the system temperature to 44°C, add 163 g of mixture 1 while stirring, and keep warm for 40 minutes. Then, perform a first heating at a rate of 2°C / min to 56°C and keep warm for 60 minutes. Then perform a second heating at a rate of 2°C / min to 94°C, keep warm for 30 minutes, and cool naturally to 65°C to obtain mixture 2; S6. 11.2 g of α-amylase with an activity of 270 U / g, 3 g of proline endoenzyme with an activity of 110 U / g, 7.2 g of saccharifying enzyme with an activity of 17.2 U / g, and 4 g of β-glucanase with an activity of 7 U / g were mixed to obtain a complex enzyme preparation; 1.3 g of the complex enzyme preparation was added to the mixture 2, and the mixture was kept warm at 65 ° C and subjected to an iodine test. After the iodine test was normal, the temperature was raised for the third time at a heating rate of 2 ° C / min, raised to 74 ° C, kept warm for 20 min, and filtered to obtain wort; S7. The wort was boiled for 82 min and 0.3 g of Simcoe hops and 0.42 g of grape leaves with a fineness of 0.28 mm were added, as well as 4 g of freeze-dried Cynanchum multiflorum powder with a fineness of 0.15 mm, wherein the Cynanchum multiflorum powder and grape leaves were added at the start of boiling, and Simcoe hops were added three times, the first time half was added at 10 min, the second time a quarter was added at 40 min, and the third time a quarter was added at 65 min; after boiling, the wort was filtered using a conventional filter tank, the tank was washed twice and the temperature of the wash water was 78 ° C, and then vortexed for 15 min, and the wort concentration was adjusted to 11 ° P after cooling, and the wort was sterilized by boiling for 30 min and then cooled to room temperature to obtain clarified wort; S8. 16.5 g of the yeast composition is added to the clarified wort, and the wort is placed in a fermenter for a primary fermentation stage. The wort is first fermented at 12° C. for 5 days. The fermentation tank is sealed when the residual sugar drops to 4° C. After 2 days, the temperature is lowered by 1° C. every 3 hours until it reaches 4° C. A secondary fermentation is carried out for 7 days until the diacetyl content is ≤ 0.05 mg / L. The fermentation is terminated, and 0.14 g of Cascade hops is dry-doped. After soaking for 5 days, the beer is filtered, bottled, and pasteurized to obtain a beer rich in Radix Polygoni Multiflori.

[0028] Comparative Example 1: Compared with Example 1, this comparative example did not add Cascade hops during the preparation of beer. The remaining steps and parameters were the same and will not be repeated in this comparative example. Finally, beer rich in Cynanchum multiflorum was obtained.

[0029] Comparative Example 2: Compared with Example 1, grape leaves were not added during the preparation of beer in this comparative example. The remaining steps and parameters were the same and will not be repeated in this comparative example. Finally, beer rich in Cynanchum multiflorum was obtained.

[0030] Comparative Example 3: Compared with Example 1, this comparative example only replaces the "complex enzyme preparation" with "saccharifying enzyme", and the remaining steps and parameters are the same, which will not be repeated in this comparative example. Finally, beer rich in Cynanchum multiflorum is obtained.

[0031] Comparative Example 4: Compared with Example 1, this comparative example only replaces the "yeast composition" with "Pichia kluyveri JLG30201", and the remaining steps and parameters are the same, which will not be repeated in this comparative example. Finally, beer rich in Cynanchum multiflorum is obtained.

[0032] Comparative Example 5: Compared with Example 1, this comparative example only replaced the "yeast composition" with "abnormal Wickham M3 yeast", and the remaining steps and parameters were the same, which will not be repeated in this comparative example. Finally, beer rich in Cynanchum multiflorum was obtained.

[0033] Comparative Example 6: Compared with Example 1, in this comparative example, no Cynanchum multiflorum powder was added during the boiling process of beer. The remaining steps and parameters were the same and will not be repeated in this comparative example. Finally, beer rich in Cynanchum multiflorum was obtained.

[0034] Comparative Example 7: S1. Weigh 10 g of yeast extract, 20 g of peptone, and 20 g of glucose into distilled water, stir until completely dissolved, dilute to 1 L, adjust the pH to 6.5, add 10 g of agar, heat at 50°C until completely dissolved, filter, and autoclave at 121°C for 15 min to obtain YPD solid medium. Weigh 10 g of yeast extract, 20 g of peptone, and 20 g of glucose into distilled water, stir until completely dissolved, dilute to 1 L, adjust the pH to 6.5, filter, and autoclave at 121°C for 15 min to obtain YPD liquid medium. S2. Pichia kluyveri JLG30201 and Wickham's M3 yeast were inoculated into YPD solid medium and cultured at 30°C for 20 hours to obtain activated strains. The strains were then inoculated into YPD liquid medium and cultured at 30°C with a shaker at 160 rpm for 20 hours to obtain primary seed solutions of Pichia kluyveri and Wickham's M3 yeast. S3. 100 mL of a Pichia kluyveri primary seed solution and 100 mL of an abnormal Wickham M3 yeast primary seed solution were inoculated into a fermenter containing 700 mL of YPD liquid medium to obtain a secondary Pichia kluyveri seed solution and an abnormal Wickham M3 yeast secondary seed solution. The secondary Pichia kluyveri seed solution and the abnormal Wickham M3 yeast secondary seed solution were mixed at an effective viable count of 2:0.5 and centrifuged at 4000 rpm for 8 min, followed by washing twice with 0.85% sterile NaCl solution to obtain a yeast composition. S4. Take the washed Cynanchum multiflorum and cut it into slices with a thickness of 3 mm. After pre-freezing at -20 ° C for 1 h, freeze-drying was performed. The temperature of the first freezing stage was -42 ° C for 5 h; the temperature of the second drying stage was -15 ° C for 15 h; then the mixture was crushed at 4 ° C to obtain a Cynanchum multiflorum powder with a fineness of 0.1 mm; 62 g of barley malt and 50 g of wheat malt were crushed by moisture regeneration method to obtain wheat flour with a fineness of 0.15 mm, which was thoroughly mixed with 28 g of Cynanchum multiflorum powder to obtain a mixture 1; S5. Add 530 mL of brewing water to the saccharification cup, and add lactic acid to adjust the pH of the system to 4. Raise the system temperature to 38°C, add 140 g of Mix 1 while stirring, and keep warm for 30 minutes. Then, perform a first heating at a rate of 1°C / min to 52°C and keep warm for 50 minutes. Then perform a second heating at a rate of 1°C / min to 90°C, keep warm for 25 minutes, and cool naturally to 62°C to obtain Mix 2. S6. 8 g of α-amylase with an activity of 220 U / g, 2.4 g of proline endoenzyme with an activity of 95 U / g, 6.5 g of saccharifying enzyme with an activity of 15.5 U / g, and 3.3 g of β-glucanase with an activity of 6 U / g were mixed to obtain a complex enzyme preparation; 0.8 g of the complex enzyme preparation was added to the mixture 2, and the mixture was kept warm at 62 ° C and iodine test was performed. After the iodine test was normal, the temperature was raised for the third time at a heating rate of 1 ° C / min, and the temperature was raised to 70 ° C, kept warm for 15 min, and filtered to obtain wort; S7. The wort was boiled for 74 minutes and 0.27g of Simcoe hops with a fineness of 0.24mm and 0.32g of grape leaves were added at the beginning of the boil, as well as 2g of freeze-dried Cynanchum multiflorum powder with a fineness of 0.1mm. After boiling, the wort was filtered using a conventional filter tank, washed twice with a water temperature of 75°C, and then vortexed for 10 minutes. After cooling, the wort concentration was adjusted to 11°P, boiled and sterilized for 20 minutes, and then cooled to room temperature to obtain clarified wort. S8. 8.2 g of the yeast composition is added to the clarified wort, and the wort is placed in a fermenter for a primary fermentation stage. The wort is first fermented at 10°C for 4 days. The fermentation tank is sealed when the residual sugar drops to 4°P. After 2 days, the temperature is lowered by 1°C every 3 hours until it drops to 2°C. A secondary fermentation is carried out for 7 days until the diacetyl content is ≤0.05 mg / L. The fermentation is terminated, and 0.1 g of Cascade hops is dry-doped into the wort. After soaking for 3 days, the beer is filtered, bottled, and pasteurized to obtain a beer rich in Radix Polygoni Multiflori.

[0035] Performance testing: Turbidity: The turbidity values ​​of the beers rich in Radix Polygoni Multiflori prepared in Examples 1 to 4 and Comparative Examples 1 to 7 were measured using a turbidimeter. Accelerated aging test: The beers rich in Radix Cynanchum multiflorum prepared in Examples 1 to 4 and Comparative Examples 1 to 7 were stored at 60° C. for 7 days to observe whether turbidity occurred. Cold turbidity test: The beers rich in Radix Cynanchum multiflorum prepared in Examples 1 to 4 and Comparative Examples 1 to 7 were refrigerated at 4° C. for 24 hours to observe whether turbidity occurred. Total acid: The total acid content is determined according to the acid-base indicator titration method in accordance with GB12456-2021 "Determination of total acid in foods"; Bitterness: Add 5 mL of filtered alcohol sample to a 50 mL centrifuge tube, then add 0.5 mL of 3 mol / L HCl solution and 10 mL of isooctane. Seal the alcohol sample and shake it on an oscillator for 15 minutes. After the solution separates, transfer the supernatant to a cuvette. Use the absorbance of isooctane at 275 nm as a blank and measure the absorbance of the alcohol sample at 275 nm. Bitterness of the wine sample to be tested (BU) = A 275 ×50 Where: A 275 is the absorbance value of the wine sample to be tested at a wavelength of 275nm; Diacetyl content: Diacetyl content is determined according to GB 4928-2008 "Analysis Method for Beer"; Sensory evaluation: A panel of 20 students and teachers who had received professional training in beer tasting evaluated the appearance, taste, flavor, and foam performance of the beers enriched with Radix Polygoni Multiflori prepared in Examples 1 to 4 and Comparative Examples 1 to 7 of the present invention. The average score was taken out of 100. The specific scoring criteria are shown in Table 1.

[0036] Table 1

[0037] Table 2

[0038] Table 3

[0039] Data Analysis: As can be seen from Tables 1, 2 and 3, the beer prepared by the present invention has higher non-biological stability, lower bitterness and better sensory evaluation. This may be due to the synergistic effect of α-amylase, proline endonuclease, saccharifying enzyme and β-glucanase in the complex enzyme preparation, which promotes saccharification, reduces the influence of the bitter substances in Radix Polygoni Multiflori on the flavor of beer, and can fully release the active substances in Radix Polygoni Multiflori, effectively exerting antioxidant and other functions; and the proline endonuclease therein, on the one hand, combines proline with polyphenols in Radix Polygoni Multiflori to form sensitive proteins, and the proline endonuclease can selectively cut off the polyphenol binding region in the sensitive protein, thereby increasing certain amino acids for the fermentation process, and can also reduce easily precipitated polypeptide fragments, thereby improving the non-biological stability of the system; on the other hand, by decomposing the proline polypeptide, the interaction force between the polypeptide and the protein is significantly reduced, thereby improving the clarity and non-biological stability of the beer; Simcoe hops are added three times during the boiling process, and while volatilizing part of the aroma, the α-acid therein isomerizes, providing basic bitterness for the beer, and during fermentation Dry hopping with Cascade hops can retain the volatile aromatic substances of hops to the maximum extent without increasing the bitter substances, and can also improve the antioxidant capacity of beer; the two hops provide basic bitterness for beer, and at the same time, they can also make the special flavor of Baishouwu compatible with the overall flavor of beer to the greatest extent possible, reduce the obvious bitterness of Baishouwu beer, and enhance the overall taste and three-dimensional level of beer; and, at the beginning of boiling, a small amount of grape leaves and Baishouwu powder are added to the wort. The bitter substances in the Baishouwu powder added for the second time can be compatible with the bitter substances in the hops. The synergistic effect of the α-acids in the white shouwu powder further enriches the taste of the beer. Moreover, adding a small amount of polyphenols in the white shouwu powder during boiling can also cooperate with the polyphenols in the hops to promote the precipitation of thermal coagulants, thereby improving the non-biological stability of the beer. Grape leaves not only have antioxidant properties, but can also form complexes with polysaccharides and other substances in the white shouwu powder to enhance the colloidal stability of the beer and improve the retention rate of the active substances in the white shouwu. In addition, the astringent feeling of the tannins contained in them may be harmonized with the bitterness in the white shouwu to enrich the taste.

[0040] Those skilled in the art should understand that the discussion of any of the above embodiments is merely illustrative and is not intended to imply that the scope of the present invention is limited to these examples. Within the scope of the present invention, the technical features in the above embodiments or different embodiments may be combined, the steps may be implemented in any order, and there are many other variations of the different aspects of the present invention as described above, which are not provided in detail for the sake of simplicity.

[0041] The present invention is intended to cover all such substitutions, modifications and variations that fall within the broad scope of the appended claims. Therefore, any omissions, modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A beer rich in Cynanchum multiflorum, characterized in that: Including the following raw materials by weight: 62-70 parts of barley malt; 50-57 parts of wheat malt; 30-40 parts of Cynanchum multiflorum; 0.32-0.42 parts of grape leaves; 0.8-1.3 parts of complex enzyme preparation; 8.2-16.5 parts of yeast composition; 0.1-0.14 parts of Cascade hops; 0.27-0.3 parts of Simcoe hops; The yeast composition is prepared from Pichia kluyveri and Wickham M3 yeast; The complex enzyme preparation is composed of α-amylase, endoproline enzyme, saccharifying enzyme and β-glucanase in a mass ratio of 8-11.2:2.4-3:6.5-7.2:3.3-4; The activity of α-amylase in the complex enzyme preparation is 220-270 U / g; The activity of the proline endo-nase in the complex enzyme preparation is 95-110 U / g; The activity of the saccharifying enzyme in the complex enzyme preparation is 15.5-17.2 U / g; The activity of β-glucanase in the complex enzyme preparation is 6-7 U / g.

2. The beer rich in Cynanchum multiflorum according to claim 1, characterized in that The preparation method of the yeast composition is as follows: Step A1. Pichia kluyveri and Wickham's M3 yeast were inoculated into YPD solid medium, cultured at 30-35°C for 20-36 hours to obtain activated strains, and then inoculated into YPD liquid medium and cultured at 30-37°C and a shaker speed of 160-180 r / min for 20-36 hours to obtain a first-level seed solution of Pichia kluyveri and a first-level seed solution of Wickham's M3 yeast; Step A2. The first-level seed liquid of Pichia kluyveri and the first-level seed liquid of Wickham's M3 yeast were respectively inoculated into a fermentation tank containing YPD liquid medium for cultivation to obtain a second-level seed liquid of Pichia kluyveri and a second-level seed liquid of Wickham's M3 yeast. The second-level seed liquid of Pichia kluyveri and the second-level seed liquid of Wickham's M3 yeast were mixed evenly at an effective viable count of 2-3:0.5-1, and centrifuged at 4000-5000 rpm for 5-8 min, and then washed twice with 0.85% sterile NaCl solution to obtain a yeast composition.

3. The beer rich in Cynanchum multiflorum according to claim 2, characterized in that The number of viable Pichia kluyveri in the secondary seed solution of Pichia kluyveri in step A2 is 3.2-4.6×10 8 cfu / mL.

4. The beer rich in Cynanchum multiflorum according to claim 2, characterized in that The volume ratio of the Pichia kluyveri primary seed solution to the YPD liquid medium in step A2 is 100:700-900; The volume ratio of the abnormal Wickham M3 yeast first-level seed liquid to the YPD liquid culture medium is 100:700-900.

5. The method for preparing beer rich in Cynanchum multiflorum according to any one of claims 1 to 4, characterized in that: The following steps are involved: Step S1. Washed radix polygoni multiflori is cut into thin slices with a thickness of 3-5 mm, pre-frozen at -20-15°C for 1-2 hours, freeze-dried, and then pulverized at 4°C to obtain radix polygoni multiflori powder with a fineness of 0.1-0.15 mm; barley malt and wheat malt are pulverized by a moisture recovery pulverization method to obtain wheat flour with a fineness of 0.15-0.2 mm, which is thoroughly mixed with the radix polygoni multiflori powder to obtain a mixture 1; Step S2. Add brewing water to the saccharification cup and add lactic acid to adjust the pH of the system to 4-5. Raise the system temperature to T1 and add Mix 1 while stirring. Keep warm for 30-40 minutes. Then, perform a first heating and keep warm for 50-60 minutes. Then, perform a second heating and keep warm for 25-30 minutes. Cool naturally to T2 to obtain Mix 2. Step S3. Add the complex enzyme preparation to the mixture 2, keep warm at temperature T2 and perform an iodine test. After the iodine test is normal, heat for the third time, keep warm for 15-20min, filter, and obtain wort; Step S4. The wort is boiled for 74-82 minutes, and Simcoe hops and grape leaves with a fineness of 0.24-0.28 mm and freeze-dried Cynanchum multiflorum powder with a fineness of 0.1-0.15 mm are added. After boiling, the wort is filtered using a conventional lauter tun, rinsed twice with the lauter tun water at a temperature of 75-78°C, and then vortexed for 10-15 minutes. After cooling, the wort concentration is adjusted to 11°P, and the wort is boiled and sterilized for 20-30 minutes and then cooled to room temperature to obtain clarified wort. Step S5. The yeast composition is added to the clarified wort, and the wort is placed in a fermenter for a primary fermentation stage. The wort is first fermented at 10-12°C for 4-5 days. The wort is sealed when the residual sugar drops to 4°C. After 2 days, the wort is cooled by 1°C every 3 hours until it drops to 2-4°C. A secondary fermentation is carried out for 7 days until the diacetyl content is ≤0.05 mg / L. The fermentation is terminated, and Cascade hops are dry-added. After soaking for 3-5 days, the beer is filtered, bottled, and pasteurized to obtain a beer rich in Radix Polygoni Multiflori.

6. The method for preparing beer rich in Cynanchum multiflorum according to claim 5, characterized in that: The freeze drying in step S1 includes a first freezing stage and a second drying stage; The temperature during the first freezing stage is -42-35°C and the time is 5-6.5 hours; The temperature during the second drying stage is -15-0°C and the time is 15-18 hours.

7. The method for preparing beer rich in Cynanchum multiflorum according to claim 5, characterized in that: The temperature T1 in step S2 is 38-44°C; The heating rate during the first heating is 1-2°C / min, and the temperature is raised to 52-56°C; The second heating process is carried out at a heating rate of 1-2°C / min to a temperature of 90-94°C.

8. The method for preparing beer rich in Cynanchum multiflorum according to claim 5, characterized in that: The temperature T2 in step S2 is 62-65°C; The ratio of brewing water to mixed material 1 is 530-680 mL:140-163 g.

9. The method for preparing beer rich in Cynanchum multiflorum according to claim 5, characterized in that: The third heating in step S3 is performed at a heating rate of 1-2°C / min to a temperature of 70-74°C.

10. The method for preparing beer rich in Cynanchum multiflorum according to claim 5, characterized in that: The Cynanchum multiflorum powder and grape leaves described in step S4 are added at the beginning of boiling; The Simcoe hops were added in three times: one-half of the hops was added at 10 min of boiling for the first time, one-quarter of the hops was added at 40 min of boiling for the second time, and one-quarter of the hops was added at 65 min of boiling for the third time.

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