Method for preparing acanthopanax beer by using pichia pastoris

Abstract

The invention discloses a method for preparing acanthopanax beer by using pichia pastoris. The acanthopanax beer is prepared through the steps of acanthopanax pretreatment, fermentation material preparation, fermentation, packaging and the like. The preparation method comprises the following steps: heating the acanthopanax in a water bath to extract main effective substances in the acanthopanax to the maximum extent, then mixing the main effective substances with saccharified and boiled wort, inoculating pichia pastoris for fermentation treatment, and fermenting by utilizing the pichia pastoris, so that on one hand, the effective components of the acanthopanax are retained to the maximum extent; on the basis of saving the cost, the effective substances in the acanthopanax senticosus are maximally fused into the beer body, and on the other hand, the fermentation of the pichia pastoris enables the beer body to be full, so that the taste of the beer is better. The richness of the acanthopanax in the edible form is expanded, meanwhile, the development of the acanthopanax and wine industry is promoted, and the acanthopanax beer has important significance on deep development and utilization of medicinal and edible resources, so that the acanthopanax beer is better in quality and has a better health-care function.

Description

Technical Field

[0001] The invention belongs to the technical field of brewing, and in particular relates to a method for preparing acanthopanax senticosus beer by utilizing Pichia yeast. Background Art

[0002] The roots, stems, leaves, flowers and fruits of Acanthopanax can all be used as medicine. As a plant with both medicinal and edible properties, Acanthopanax has an important position in traditional Chinese medicine and food culture. It can be used as a medicinal material or as a food ingredient. According to the latest version of the list of medicinal and edible substances released on July 23, 2024, Acanthopanax was clearly listed as a medicinal and edible substance. The tender stems and leaves of Acanthopanax are edible, with a unique flavor, a fresh and slightly bitter fragrance, and can be used for stir-frying, cold salad, stuffing, and soup. Its leaves can also be made into Acanthopanax tea, which has a special aroma and anti-fatigue effect. In addition, the fruits of Acanthopanax can be used to make wine or extract oil. The roots, rhizomes or stems of Acanthopanax are used as medicine, which have the effects of invigorating qi and strengthening the spleen, nourishing the kidneys and calming the mind. It is mainly used to treat symptoms such as spleen and lung qi deficiency, physical weakness, loss of appetite, lung and kidney deficiency, long-term cough and asthma, kidney deficiency, waist and knee pain, heart and spleen deficiency, insomnia and dreaminess. Acanthopanax mainly contains a variety of glycosides, such as eleutheroside, syringin, lirioside, etc.; coumarin components, such as isoflavone, etc.; lignans, such as sesame, etc. It also contains sugars, fatty acids and quinones. Modern research shows that the active ingredients in Acanthopanax can weaken the inhibition of chloral hydrate, sodium barbital and chlorpromazine on the central nervous system, and increase the phagocytic ability of mononuclear phagocytes and peritoneal macrophages. In addition, it also has sedative, anti-fatigue, hypoxia resistance, anti-stress, detoxification, anti-aging, antibody production promotion, interferon production promotion, anti-inflammatory, anti-platelet aggregation, antibacterial, antiviral and other effects.

[0003] Currently, there are few studies on the use of Acanthopanax senticosus to ferment beer, and the main method is to make blended wine by soaking. However, there are few studies on Acanthopanax senticosus fermented wine, especially fermented beer, which lacks aroma and has a poor taste. Summary of the invention

[0004] The present invention aims to overcome the above problems and provide a method for preparing Acanthopanax senticosus beer by using Pichia yeast, which can improve the utilization rate of the effective ingredients of Acanthopanax senticosus and further give play to the role of Pichia yeast in producing wine and phenylethyl acetate, so that the quality of Acanthopanax senticosus beer is better and has more health care function.

[0005] A method for preparing Acanthopanax senticosus beer by utilizing Pichia yeast of the present invention comprises the following steps: (1) Pretreatment of Acanthopanax senticosus: Crush the Acanthopanax senticosus and sieve through a 20-mesh sieve. Add water to the Acanthopanax senticosus at a material-water ratio of 5% by mass, keep the mixture at 18-22°C for 28-32 minutes, and filter the mixture to obtain the filtrate. (2) Malt crushing: Use a grinder to crush the malt. The malt crushing requires that the bran should be broken but not shattered, and should be as complete as possible. The white endosperm material should be crushed as completely as possible. (3) Saccharification and filtration: Step-by-step saccharification: weigh the crushed malt, put it into a filter bag, and mix it with water in a saccharification pot at a mass ratio of 1:5, where the pH of the water is required to be 5.4. Then adjust the temperature to 45°C and keep it warm for 60 minutes for the first step of saccharification; then slowly raise the temperature to 67°C and continue to keep it warm for 60 minutes for the second step of saccharification, until the saccharification liquid no longer shows blue when tested with iodine solution, and finally the saccharification is completed; (3) Boiling and adding hops: put the filtered wort into the boiling pot and keep it at a slight boil for 1 hour. During the heating process, add granulated hops three times. The amount of granulated hops added is 0.1% of the wort mass. The hops are added in a three-step method. The first addition is made when the wort is just heated, and the amount added is 20% of the total amount of hops; the second addition is made after 40 minutes of boiling, and the amount added is 50% of the total amount of hops; the third addition is made after 55 minutes of boiling, and the amount added is 30% of the total amount of hops. Finally, boil for another 5 minutes, and the boiling is completed; (4) Filtration and clarification: Filter and clarify the wort; (5) Main fermentation: The boiled wort was rapidly cooled to 23-27°C, and mixed with the Acanthopanax senticosus treatment solution at a volume ratio of 3:1 to obtain a fermentation material; Pichia kudriavzevii (FBKL2.0008) was activated and cultured, and the activated Pichia kudriavzevii was inoculated at 7-9% of the volume of the fermentation material, and the main fermentation was carried out at a constant temperature of 18°C ​​for 10 days; (6) Post-fermentation: After the main fermentation is completed, the temperature is transferred to 10°C for post-fermentation, which takes about 15 days; (7) Packaging: After filtering the fermented liquid, packaging and pasteurization are performed to obtain Acanthopanax senticosus beer; (8) Physical and chemical index detection: alcohol content 3.3-3.7%vol, total acid: 2.2-2.8ml / 100ml, carbon dioxide 0.35-0.65%vol, diacetyl 0.01-0.093mg / L.

[0006] The above method for preparing Acanthopanax senticosus beer using Pichia yeast, wherein the Pichia yeast comes from China Typical Industrial Microbiological Culture Collection Center, with a collection number of CCTCC M 2015380 and a name of Pichia kudriavzevii FBKL2.0008.

[0007] In the above method for preparing Acanthopanax senticosus beer using Pichia yeast, the activation culture in step (5) is to activate Pichia yeast FBKL2.0008 (Pichia kudriavzevii, FBKL2.0008) on a solid slant medium of malt juice for 2-3 days, then inoculate it into a liquid malt juice medium and culture it at 28°C for 2 days.

[0008] Compared with the prior art, the present invention heats Acanthopanax senticosus in a water bath to extract the effective substances in Acanthopanax senticosus to the maximum extent, and then mixes with the wort after saccharification and boiling, and inoculates with Pichia yeast for fermentation. Pichia kudriavzevii FBKL2.0008 (Pichia kudriavzevii, FBKL2.0008) is derived from the fermented grains of Guizhou traditional Maotai-flavor liquor, has a strong wine-producing ability, belongs to non-wine yeast, has the ability to produce phenylethyl acetate, and is the main source of sweet aroma in the wine. Phenylethyl acetate is sweet, rose-like, with a powdery honey-like aroma, similar to apple-like fruity aroma, and has a cocoa and whiskey-like aroma, which can better neutralize the bitterness brought by Acanthopanax senticosus in the wine. This method retains the effective substances of Acanthopanax senticosus to the greatest extent on the one hand, and on the other hand, the fermentation of Pichia yeast makes the wine full and the sweet aroma coordinated. The invention not only expands the richness of Acanthopanax senticosus in edible forms, but also promotes the development of Acanthopanax senticosus and wine industry, and has important significance for the deep development and utilization of medicine and food resources. The invention can not only improve the utilization rate of the effective ingredients of Acanthopanax senticosus, but also further play the role of Pichia yeast in producing wine and phenylethyl acetate, so that the quality of Acanthopanax senticosus beer is better and has more health care functions. DETAILED DESCRIPTION Example 1

[0009] A method for preparing Acanthopanax senticosus beer using Pichia yeast comprises the following steps: (1) Pretreatment of Acanthopanax senticosus: Crush the Acanthopanax senticosus and sieve through a 20-mesh sieve. Add water to the Acanthopanax senticosus at a material-water ratio of 5% by mass, keep the mixture at 18-22°C for 28-32 minutes, and filter the mixture to obtain the filtrate. (2) Malt crushing: Use a grinder to crush the malt. The malt crushing requires that the bran should be broken but not shattered, and should be as complete as possible. The white endosperm material should be crushed as completely as possible. (3) Saccharification and filtration: Step-by-step saccharification: weigh the crushed malt, put it into a filter bag, and mix it with water in a saccharification pot at a mass ratio of 1:5, where the pH of the water is required to be 5.4. Then adjust the temperature to 45°C and keep it warm for 60 minutes for the first step of saccharification; then slowly raise the temperature to 67°C and continue to keep it warm for 60 minutes for the second step of saccharification, until the saccharification liquid no longer shows blue when tested with iodine solution, and finally the saccharification is completed; (3) Boiling and adding hops: put the filtered wort into the boiling pot and keep it at a slight boil for 1 hour. During the heating process, add granulated hops three times. The amount of granulated hops added is 0.1% of the wort mass. The hops are added in a three-step method. The first addition is made when the wort is just heated, and the amount added is 20% of the total amount of hops; the second addition is made after 40 minutes of boiling, and the amount added is 50% of the total amount of hops; the third addition is made after 55 minutes of boiling, and the amount added is 30% of the total amount of hops. Finally, boil for another 5 minutes, and the boiling is completed; (4) Filtration and clarification: Filter and clarify the wort; (5) Main fermentation: The boiled wort was rapidly cooled to about 25°C, and mixed with the Acanthopanax senticosus treatment solution at a volume ratio of 3:1 to obtain a fermentation material; Pichia kudriavzevii (FBKL2.0008) was activated and cultured, and the activated Pichia kudriavzevii was inoculated at 7% of the volume of the fermentation material, and the main fermentation was carried out at a constant temperature of 18°C ​​for 10 days; (6) Post-fermentation: After the main fermentation is completed, the temperature is transferred to 10°C for post-fermentation, which takes about 15 days; (7) Packaging: After filtering the fermented liquid, packaging and pasteurization are performed to obtain Acanthopanax senticosus beer; (8) Physical and chemical index detection: alcohol content 3.3%vol, total acid: 2.2ml / 100ml, carbon dioxide 0.35%vol, diacetyl 0.01mg / L. Example 2

[0010] A method for preparing Acanthopanax senticosus beer using Pichia yeast comprises the following steps: (1) Pretreatment of Acanthopanax senticosus: Crush the Acanthopanax senticosus and sieve through a 20-mesh sieve. Add water to the Acanthopanax senticosus at a material-water ratio of 5% by mass, keep the mixture at 18-22°C for 28-32 minutes, and filter the mixture to obtain the filtrate. (2) Malt crushing: Use a grinder to crush the malt. The malt crushing requires that the bran should be broken but not shattered, and should be as complete as possible. The white endosperm material should be crushed as completely as possible. (3) Saccharification and filtration: Step-by-step saccharification: weigh the crushed malt, put it into a filter bag, and mix it with water in a saccharification pot at a mass ratio of 1:5, where the pH of the water is required to be 5.4. Then adjust the temperature to 45°C and keep it warm for 60 minutes for the first step of saccharification; then slowly raise the temperature to 67°C and continue to keep it warm for 60 minutes for the second step of saccharification, until the saccharification liquid no longer shows blue when tested with iodine solution, and finally the saccharification is completed; (3) Boiling and adding hops: put the filtered wort into the boiling pot and keep it at a slight boil for 1 hour. During the heating process, add granulated hops three times. The amount of granulated hops added is 0.1% of the wort mass. The hops are added in a three-step method. The first addition is made when the wort is just heated, and the amount added is 20% of the total amount of hops; the second addition is made after 40 minutes of boiling, and the amount added is 50% of the total amount of hops; the third addition is made after 55 minutes of boiling, and the amount added is 30% of the total amount of hops. Finally, boil for another 5 minutes, and the boiling is completed; (4) Filtration and clarification: Filter and clarify the wort; (5) Main fermentation: The boiled wort was rapidly cooled to about 25°C, and mixed with the Acanthopanax senticosus treatment solution at a volume ratio of 3:1 to obtain a fermentation material; Pichia kudriavzevii (FBKL2.0008) was activated and cultured, and the activated Pichia kudriavzevii was inoculated at 8% of the volume of the fermentation material, and the main fermentation was carried out at a constant temperature of 18°C ​​for 10 days; (6) Post-fermentation: After the main fermentation is completed, the temperature is transferred to 10°C for post-fermentation, which takes about 15 days; (7) Packaging: After filtering the fermented liquid, packaging and pasteurization are performed to obtain Acanthopanax senticosus beer; (8) Physical and chemical index detection: alcohol content 3.5%vol, total acid: 2.4ml / 100ml, carbon dioxide 0.58%vol, diacetyl 0.052mg / L. Example 3

[0011] A method for preparing Acanthopanax senticosus beer using Pichia yeast comprises the following steps: (1) Pretreatment of Acanthopanax senticosus: Crush the Acanthopanax senticosus and sieve through a 20-mesh sieve. Add water to the Acanthopanax senticosus at a material-water ratio of 5% by mass, keep the mixture at 18-22°C for 28-32 minutes, and filter the mixture to obtain the filtrate. (2) Malt crushing: Use a grinder to crush the malt. The malt crushing requires that the bran should be broken but not shattered, and should be as complete as possible. The white endosperm material should be crushed as completely as possible. (3) Saccharification and filtration: Step-by-step saccharification: weigh the crushed malt, put it into a filter bag, and mix it with water in a saccharification pot at a mass ratio of 1:5, where the pH of the water is required to be 5.4. Then adjust the temperature to 45°C and keep it warm for 60 minutes for the first step of saccharification; then slowly raise the temperature to 67°C and continue to keep it warm for 60 minutes for the second step of saccharification, until the saccharification liquid no longer shows blue when tested with iodine solution, and finally the saccharification is completed; (3) Boiling and adding hops: put the filtered wort into the boiling pot and keep it at a slight boil for 1 hour. During the heating process, add granulated hops three times. The amount of granulated hops added is 0.1% of the wort mass. The hops are added in a three-step method. The first addition is made when the wort is just heated, and the amount added is 20% of the total amount of hops; the second addition is made after 40 minutes of boiling, and the amount added is 50% of the total amount of hops; the third addition is made after 55 minutes of boiling, and the amount added is 30% of the total amount of hops. Finally, boil for another 5 minutes, and the boiling is completed; (4) Filtration and clarification: Filter and clarify the wort; (5) Main fermentation: The boiled wort was rapidly cooled to about 25°C, and mixed with the Acanthopanax senticosus treatment solution at a volume ratio of 3:1 to obtain a fermentation material; Pichia kudriavzevii (FBKL2.0008) was activated and cultured, and the activated Pichia kudriavzevii was inoculated at 9% of the volume of the fermentation material, and the main fermentation was carried out at a constant temperature of 18°C ​​for 10 days; (6) Post-fermentation: After the main fermentation is completed, the temperature is transferred to 10°C for post-fermentation, which takes about 15 days; (7) Packaging: After filtering the fermented liquid, packaging and pasteurization are performed to obtain Acanthopanax senticosus beer; (8) Physical and chemical index detection: alcohol content 3.7%vol, total acid: 2.8ml / 100ml, carbon dioxide 0.65%vol, diethyl 0.093mg / L.

[0012] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any form. Any simple modification, equivalent change and modification made to the above embodiment according to the technical essence of the present invention without departing from the technical solution of the present invention still falls within the scope of the technical solution of the present invention.

Claims

1. A method for preparing Acanthopanax senticosus beer using Pichia yeast, characterized in that The following steps are involved: (1) Pretreatment of Acanthopanax senticosus: Crush the Acanthopanax senticosus and sieve through a 20-mesh sieve. Add water to the Acanthopanax senticosus at a material-water ratio of 5% by mass, keep the mixture at 18-22°C for 28-32 minutes, and filter the mixture to obtain the filtrate. (2) Malt crushing: Use a grinder to crush the malt. The malt crushing requires that the bran is broken but not shattered, and the white endosperm material is required to be crushed as completely as possible; (3) Saccharification and filtration: Step-by-step saccharification: Weigh the crushed malt and mix it with water in a saccharification pot at a weight ratio of 1:5, where the pH of the water is required to be 5.

4. Then adjust the temperature to 45°C and let it stand for 60 minutes for the first step of saccharification; then slowly raise the temperature to 67°C and continue to stand for 60 minutes for the second step of saccharification until the saccharification liquid no longer shows blue when tested with iodine solution. Finally, saccharification is completed; (3) Boiling and adding hops: put the filtered wort into the boiling pot and keep it at a slight boil for 1 hour. During the heating process, add granulated hops three times. The amount of granulated hops added is 0.1% of the weight of the wort. The hops are added in three steps. The first addition is made when the wort is just heated, and the amount added is 20% of the total amount of hops; the second addition is made after 40 minutes of boiling, and the amount added is 50% of the total amount of hops; the third addition is made after 55 minutes of boiling, and the amount added is 30% of the total amount of hops. Finally, boil for 5 minutes and the boiling is completed. (4) Filtration and clarification: Filter and clarify the wort; (5) Main fermentation: The boiled wort was rapidly cooled to 23-27°C and mixed with the Acanthopanax senticosus treatment solution at a volume ratio of 3:1 to obtain a fermentation material; Pichia yeast FBKL2.0008 ( Pichia kudriavzevii , FBKL2.0008) activation culture, inoculate activated Pichia yeast at 7-9% of the volume of the fermentation material, and carry out the main fermentation at a constant temperature of 18°C ​​for 10 days; (6) Post-fermentation: After the main fermentation is completed, the temperature is transferred to 10°C for post-fermentation, which takes about 15 days; (7) Packaging: The fermented liquid is filtered, packaged, and pasteurized to obtain Acanthopanax senticosus beer.

2. A method for preparing Acanthopanax senticosus beer using Pichia yeast as claimed in claim 1, characterized in that: Pichia pastoris is from China Typical Industrial Microbial Culture Collection Center, with the collection number: CCTCC M 2015380, and the name: Pichia pastoris FBKL2.0008 ( Pichia kudriavzevii , FBKL2.0008).

3. The method for preparing Acanthopanax senticosus beer using Pichia yeast as claimed in claim 1, characterized in that: The activation culture described in step (5) is to culture Pichia pastoris FBKL2.0008 ( Pichia kudriavzevii , FBKL2.0008) was activated and cultured on malt wort solid slant medium for 2-3 days, then inoculated into malt wort liquid medium and cultured at 28°C for 2 days.