CN112159739A - Low-purine beer with uric acid reducing effect - Google Patents

Low-purine beer with uric acid reducing effect Download PDF

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CN112159739A
CN112159739A CN202011104740.1A CN202011104740A CN112159739A CN 112159739 A CN112159739 A CN 112159739A CN 202011104740 A CN202011104740 A CN 202011104740A CN 112159739 A CN112159739 A CN 112159739A
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beer
chitosan
uric acid
mass
heating
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陈列欢
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Guangzhou Youlan Marine Biotechnology Co ltd
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Guangzhou Youlan Marine Biotechnology Co ltd
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Abstract

The invention belongs to the field of health food and functional food, and discloses low-purine beer with a uric acid reducing effect, which contains marine bioactive substances such as chitosan oligosaccharide, marine fish oligopeptide, brown algae oligosaccharide and the like, can reduce the generation of uric acid in vivo and accelerate the metabolism of uric acid, and simultaneously the purine content of the beer is less than 20mg/L, so that the purine absorption of organisms is reduced from the source, and the incidence rate of gout is reduced. The invention also provides a preparation method of the low-purine beer with the effect of reducing uric acid, wherein chitosan-based graphene carbon spheres are used as an adsorbent, and the adsorbent has chemical and physical adsorption effects, so that the adsorption effect on purine in beer is obviously enhanced.

Description

Low-purine beer with uric acid reducing effect
Technical Field
The invention relates to the field of health food and functional food, in particular to low-purine beer with a uric acid reducing effect.
Background
Beer is one of the oldest alcoholic beverages in humans, the third beverage in the world after water and tea. The beer is low-alcohol wine rich in carbon dioxide, called as 'liquid bread', and is a low-concentration alcoholic beverage, which is brewed by using barley malt, hops and water as main raw materials through yeast fermentation. The purine content of domestic beer is generally 38-151 mg/L, although the purine content in the beer is moderate, the consumption amount is large, and a large amount of free purine is taken by a human body through drinking the beer, so that purine is accumulated in the body and quickly generates uric acid, the hyperuricemia is caused, and the incidence rate of gout is increased finally.
In the prior art, the risk of gout caused by drinking beer is reduced mainly by a mode of preparing low-purine beer, for example, in patent CN 101948719B, the usage amount of barley malt is reduced, the usage amount of wheat malt with relatively low purine content is increased, a special malt capable of adjusting the aroma chromaticity of the beer, namely a scorch-aroma malt, is properly adopted, nitrogen-containing grain auxiliary materials with certain purine content are abandoned, and almost purine-free corn starch syrup is completely used, so that the content of purine substances in the finished product beer is obviously reduced; in the patent CN 102311897B, a malt peeling and bud removing technology is adopted in saccharification, nucleic acid macromolecular substances in wort are reduced, malt roots are utilized in fermentation to degrade residual nucleic acid substances, and finally the content of purine substances in beer is controlled to be 10.5mg/L, but pure low-purine beer has no effect of relieving gout.
Patent CN 109161449A provides a roxburgh rose beer, which does not use malt for fermentation, simultaneously filters yeast in the beer to cut off the source of purine, utilizes an antioxidant active substance in roxburgh rose to hydrolyze oxidase which can oxidize purine into uric acid, blocks an enzyme preparation required for synthesizing uric acid, and simultaneously utilizes Guizhou characteristic Chinese herbal medicines to promote the excretion of uric acid from the body. Patent CN 107365647 a provides a pu' er tea beer, which utilizes PTP in tea polyphenol to activate the activity of antioxidant enzyme and reduce the accumulation of free radicals in human body, so as to achieve the purpose of suppressing or reducing gout attack, but the patent does not treat purine in beer, so that the effect of alleviating gout by using the antioxidant effect of tea polyphenol alone is not obvious.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention mainly aims to provide the low-purine beer with the efficacy of reducing uric acid and the malt fragrance.
The invention also aims to provide a preparation method of the low-purine beer with the effect of reducing uric acid.
In order to achieve the purpose, the invention adopts the following technical scheme:
a low-purine beer with uric acid reducing effect comprises the following active substances: chitosan oligosaccharide, marine fish oligopeptide and brown algae oligosaccharide.
Preferably, the content of the chitosan oligosaccharide in the beer is 0.01-1 g/L.
Preferably, the molecular weight of the chitosan oligosaccharide is 300-3000 Da, and more preferably 2000-3000 Da.
Preferably, the preparation method of the chitosan oligosaccharide specifically comprises the following steps:
A. adding chitosan into water, uniformly stirring, continuously adding concentrated hydrochloric acid, accelerating stirring to prepare a chitosan solution, adjusting the pH value of the chitosan solution to 5-6 by using 0.2mol/L acetic acid-sodium acetate buffer solution, and keeping the temperature of the chitosan solution at 36-40 ℃;
B. and C, adding chitosan enzyme into the chitosan solution obtained in the step A, stirring, carrying out hydrolysis reaction for 12-24 hours to obtain an enzymatic hydrolysate, heating at 90-100 ℃ for 10-20 min to inactivate enzyme, carrying out centrifugal separation, taking supernate, concentrating, and carrying out freeze drying to obtain the chitosan oligosaccharide.
Preferably, the content of chitosan in water in the step A is 3-8 wt%.
Preferably, the ratio of the concentrated hydrochloric acid to the water in the step A is 2: 100-4: 100.
Preferably, the mass ratio of the chitosan enzyme to the chitosan solution in the step B is 3: 100-6: 100.
Preferably, the content of the marine fish oligopeptide in the beer is 0.01-5 g/L.
Preferably, the marine fish oligopeptide is extracted from marine fish such as bonito and tuna.
Preferably, the marine fish oligopeptide mainly consists of dipeptide and tripeptide, and the molecular weight of the marine fish oligopeptide is less than 1000 Da.
Preferably, the marine fish oligopeptide is anserine, and the purity of the anserine is 10-90 wt%.
Preferably, the content of the brown algae oligosaccharide in the beer is 0.01-5 g/L.
Preferably, the brown algae oligosaccharide is extracted from brown algae such as kelp, Undaria pinnatifida, Hizikia fusiforme and Sargassum horneri.
Preferably, the molecular weight of the brown algae oligosaccharide is 500-3000 Da, and more preferably 2000-3000 Da.
Preferably, the preparation method of the brown algae oligosaccharide specifically comprises the following steps:
a. crushing brown algae, adding water, leaching for 2-4 h at 50-100 ℃, centrifuging to obtain supernatant, concentrating, precipitating with ethanol, refrigerating at 4 ℃ overnight, filtering, and drying to obtain crude seaweed polysaccharide;
b. deproteinizing and decolorizing the crude brown algae polysaccharide, and drying to obtain brown algae polysaccharide;
c. dissolving fucoidan in water, adjusting the pH value to 6.5-8.0, adding alginate lyase, reacting for 1-3 h at 36-40 ℃ to obtain an enzymatic hydrolysate, heating at 90-100 ℃ for 10-20 min to inactivate enzyme, performing centrifugal separation, taking supernatant, concentrating, and freeze-drying to obtain the alginate oligosaccharide.
Preferably, the brown algae is crushed, sieved by a 80-mesh sieve, and added with water for leaching.
Preferably, the ratio of the brown algae to water in the step a is 1: 10-1: 30.
Preferably, the alcohol precipitation in the step a is performed by adopting 90-95% ethanol, and the volume of the ethanol added is 2-4 times of that of the concentrated solution.
Preferably, the brown algae crude polysaccharide in step a can be washed with absolute ethyl alcohol and acetone in sequence and then dried in vacuum.
Preferably, the deproteinization in the step b adopts a Sevag method, which specifically comprises the following steps: and (b) adding water to the brown algae crude polysaccharide obtained in the step a for dissolving, adding 1/3 volumes of chloroform-n-butanol (3:1, V/V) into the crude polysaccharide solution, oscillating for 30min, centrifuging, taking supernate, and repeating for 6 times.
Preferably, the decoloring in the step b specifically comprises the following steps: and adding 2-3 wt% of activated carbon into the deproteinized crude polysaccharide solution, and decoloring for 1.5-2 h at the temperature of 50-55 ℃.
Preferably, in the step c, the concentration of the brown algae polysaccharide solution is 0.15 to 0.25 wt%.
Preferably, in the step c, the addition amount of the alginate lyase is 0.3-1U/mL.
Furthermore, other active substances, such as probiotics, apigenin, curcumin, luteolin, allicin, agaricus bisporus extract, artichoke leaf extract, artichoke extract, sour cherry extract, grape seed extract, wild vine extract, dogwood extract and the like can be added into the low-purine beer with the effect of reducing uric acid provided by the invention according to the formula requirements.
The invention also provides a preparation method of the low purine beer with the function of reducing uric acid, which comprises the following steps:
s1, respectively removing seed coats and germs of clean 55-60 parts by mass of barley malt, 25-40 parts by mass of wheat malt and 5-10 parts by mass of burnt malt, adding water to moisten wheat for 5-10 min, and then crushing the raw materials;
s2, preheating water in a saccharifying tank to 36-38 ℃, then putting the barley malt powder, the wheat malt powder and the burnt wheat malt powder which are obtained by crushing in S1 into the saccharifying tank, uniformly stirring, standing for 15-20 min, then heating to 52-53 ℃ at the speed of 1 ℃/min under the stirring condition, standing for 55-60 min, heating to 66-67 ℃ at the speed of 1 ℃/min under the stirring condition, standing for 55-60 min, and heating to 77-78 ℃ at the speed of 1 ℃/min under the stirring condition;
s3, carrying out first filtration on the saccharified mash to obtain clear wort;
s4 heating and boiling the wort, adding 0.00020-0.00025 parts by mass of bitter hops, adding 0.00015-0.00018 parts by mass of fragrant hops after 42-45 min, and stopping heating after 48-50 min;
s5, carrying out second filtration on the boiled wort while the wort is hot, removing filter residues, and cooling to 10-10.5 ℃;
s6, oxygenating the cooled wort, feeding the cooled wort into a fermentation tank, adding 0.00022-0.00025 parts by mass of yeast, fermenting under the conditions of 0.02 Mpa-0.03 MPa and 10-10.5 ℃ until the sugar degree is 5-5.5 ℃, heating to 12-12.5 ℃, continuing to ferment to 4-4.5 ℃, boosting to 0.11-0.12 MPa, keeping for 5-6 days, cooling to 5-5.5 ℃, keeping for 1 day, cooling to-1-0 ℃ and curing for 4-5 days;
s7, performing third filtration after the wine liquid is mature, removing yeast, heating to 20-25 ℃, then adding chitosan-based graphene carbon spheres, adsorbing for 1h, and performing fourth filtration;
s8 adding chitosan oligosaccharide, marine fish oligopeptide and brown algae oligosaccharide into the liquor, stirring to dissolve, and storing at-1-0 ℃.
Preferably, in the step S2, the ratio of the total mass of the barley malt flour, the wheat malt flour and the burnt malt flour to the mass of water is 4: 1-5: 1.
Preferably, the yeast is high activity brewer's yeast.
Preferably, the addition amount of the chitosan-based graphene carbon spheres is 0.02-0.05 wt% of the wine.
Preferably, the chitosan-based graphene carbon spheres are prepared by the following steps: dissolving chitosan in 1.5-2% acetic acid aqueous solution, continuously adding graphene powder, stirring uniformly, slowly dripping chitosan-graphene suspension into 2-3 mol/L NaOH solution to prepare balls, filtering out solid pellets after 2-3 h, washing with water to be neutral, adding the solid pellets into 25-30% glutaraldehyde solution for crosslinking, taking out after 3h of crosslinking, drying, and carbonizing the dried pellets at 400-420 ℃ for 0.5-0.8 h under the condition of inert atmosphere. And cooling to room temperature, washing away impurities with water, and drying to obtain the chitosan-based graphene carbon spheres.
Preferably, the mass ratio of the chitosan to the graphene is 1: 1-1: 3.
Preferably, the content of chitosan in the chitosan-graphene suspension is 2.5-3.0 wt%.
Preferably, the mass ratio of the solid globules to the glutaraldehyde solution is 1: 1-1: 2.
Preferably, the drying is drying at 80 ℃ for 10 h.
Compared with the prior art, the invention has the following advantages and technical effects:
1. the marine organism has an extreme growth environment, compared with terrestrial organisms, the bioactive substances of the marine organism show unique advantages and characteristics, and chitosan oligosaccharide, brown alga oligosaccharide and marine fish oligopeptide are small-molecular marine bioactive substances added into beer, are easily absorbed by organisms and have obvious bioactive functions, and the specific action mechanism is as follows: 1) the chitosan oligosaccharide can inhibit xanthine oxidase and reduce the in vivo generation of uric acid; 2) the marine fish oligopeptide can reduce the deposition of urate substances in kidney and accelerate uric acid metabolism; 3) the brown algae oligosaccharide has obvious antioxidation and neuroprotection effects, and can prevent alcoholic nerve injury caused by large-scale drinking; 4) the chitosan oligosaccharide and the brown algae oligosaccharide are used as prebiotics, so that the uric acid reducing effect is achieved, the intestinal flora can be adjusted, and the gastrointestinal health after drinking is guaranteed.
2. The purine content of the low-purine beer prepared by the invention is controlled to be below 20mg/L, which is 1/8-1/2 of that of common beer, so that the total amount of purine absorbed by organisms is reduced from the source, and the gout incidence rate is reduced.
3. According to the invention, chitosan-based graphene carbon spheres are used as an adsorbent, the adsorbent has good adsorption performance on purine, chitosan is used as a natural high polymer material, lone pair electrons and empty orbitals exist on amino and hydroxyl on molecules, cations formed by protonation of the amino are very easy to be subjected to electrostatic binding with anions, so that an adsorption phenomenon is shown, the specific surface area of graphene is large, the adsorption capacity is very strong, purine substances are physically adsorbed, the prepared chitosan-based graphene carbon spheres can combine chemical adsorption and physical adsorption, the purine adsorption capacity is obviously enhanced, after the carbon spheres are prepared, the chitosan also has physical adsorption performance, and after the chitosan is crosslinked with graphene, the stability of the carbon spheres is improved.
Detailed Description
The present invention will be described in further detail with reference to specific examples, but the embodiments of the present invention are not limited thereto. All the raw materials and reagents used in the present invention are commercially available raw materials and reagents, unless otherwise specified. In the examples, the components are used in g and mL in parts by mass.
For convenience, in the following examples of the present invention, "low purine beer having uric acid lowering effect" of the present invention is simply referred to as "tequilon".
Preparation of chitosan oligosaccharide:
adding 5 parts by mass of chitosan into 100 parts by volume of water, uniformly stirring, continuously adding 3 parts by mass of concentrated hydrochloric acid, accelerating stirring to prepare a chitosan solution, adjusting the pH value of the chitosan solution to 5 by using 0.2mol/L acetic acid-sodium acetate buffer solution, and keeping the temperature of the chitosan solution at 36 ℃;
adding 5 parts by mass of chitosan enzyme into the chitosan solution, stirring, carrying out hydrolysis reaction for 15 hours to obtain an enzymatic hydrolysate, heating at 90 ℃ for 20min to inactivate enzyme, carrying out centrifugal separation, taking supernatant, concentrating, and carrying out freeze drying to obtain chitosan oligosaccharide for later use.
Dissolving chitosan oligosaccharide in water, and sequentially performing dialysis with dialysis membranes with molecular cut-off amounts of 2000 and 3000 to finally obtain the chitosan oligosaccharide with molecular weight of 2000-3000 Da for later use.
Preparation of brown algae oligosaccharide:
polysaccharide extraction: crushing 1 part by mass of kelp, adding the crushed kelp into 20 parts by volume of water, leaching for 4 hours at the temperature of 80 ℃, centrifuging to obtain a supernatant, concentrating, precipitating with ethanol, refrigerating overnight at the temperature of 4 ℃, filtering, washing with absolute ethyl alcohol and acetone in sequence, and drying in vacuum to obtain brown algae crude polysaccharide;
deproteinization: dissolving brown algae crude polysaccharide in water, adding 1/3 volume of chloroform-n-butanol (3:1, V/V) into the crude polysaccharide solution, oscillating for 30min, centrifuging, collecting supernatant, and repeating for 6 times;
and (3) decoloring: adding 2 wt% of activated carbon into the deproteinized crude polysaccharide solution, decoloring for 2h at 50 ℃, filtering and drying to obtain the brown algae polysaccharide.
And (3) degradation: adding 2 parts by mass of fucoidan into 100 parts by volume of water, adjusting the pH value of the solution to 7.0, adding alginate lyase (controlling the adding amount of the alginate lyase to be 0.5U/mL), reacting for 3 hours at 40 ℃ to obtain an enzymolysis solution, heating at 90 ℃ for 15min to inactivate the enzyme, performing centrifugal separation, taking the supernatant, concentrating, and performing freeze drying to obtain the fucoidan oligosaccharide for later use.
And (3) dialysis: dissolving the brown algae oligosaccharide in water, and sequentially carrying out detection by using dialysis membranes with molecular cut-off amounts of 2000 and 3000 to finally obtain the brown algae oligosaccharide with the molecular weight of 2000-3000 Da for later use.
Preparing chitosan-based graphene carbon spheres:
dissolving 3 parts by mass of chitosan in 100 parts by mass of 2% acetic acid aqueous solution, continuously adding 4 parts by mass of graphene powder, stirring uniformly, slowly dripping the chitosan-graphene suspension into 2mol/L NaOH solution to prepare balls, filtering out solid spheres after 3h, washing with water to be neutral, adding 1 part by mass of the solid spheres into 2 parts by mass of 25% glutaraldehyde solution for crosslinking, taking out after 3h of crosslinking, drying at 80 ℃ for 10h, and carbonizing the dried spheres at 400 ℃ for 0.5h under the inert atmosphere condition. And cooling to room temperature, washing impurities with water, and drying at 80 ℃ for 10h to obtain the chitosan-based graphene carbon spheres.
Example 1: preparation of Dipiaoling beer
S1, respectively removing seed coats and germs of 55 parts by mass of clean barley malt, 25 parts by mass of wheat malt and 5 parts by mass of burnt malt, adding water to moisten wheat for 5min, and then crushing the raw materials;
s2, preheating water in a saccharifying tank to 36 ℃, putting barley malt flour, wheat malt flour and burnt malt flour into the saccharifying tank, uniformly stirring, standing for 15min, heating to 52 ℃ at the speed of 1 ℃/min under the stirring condition, standing for 55min, heating to 66 ℃ at the speed of 1 ℃/min under the stirring condition, standing for 55min, and heating to 77 ℃ at the speed of 1 ℃/min under the stirring condition;
s3, carrying out first filtration on the saccharified mash to obtain clear wort;
s4 heating and boiling the wort, adding 0.00020 parts by mass of bitter hops, adding 0.00015 parts by mass of fragrant hops after 42min, and stopping heating after 48-50 min;
s5, carrying out second filtration on the boiled wort while the wort is hot, removing filter residues, and cooling to 10-10.5 ℃;
s6, oxygenating the cooled wort, feeding the cooled wort into a fermentation tank, adding 0.00022 parts by mass of yeast, fermenting under the conditions of 0.02Mpa and 10 ℃ until the sugar degree is 5 ℃, heating to 12 ℃, continuing to ferment to 4 ℃, boosting to 0.11MPa, keeping for 5 days, cooling to 5 ℃, keeping for 1 day, cooling to-1 ℃ and curing for 4 days;
s7, performing third filtration after the wine liquid is mature, removing yeast, heating to 20 ℃, then adding 0.02 wt% of chitosan-based graphene carbon spheres, adsorbing for 1h, and performing fourth filtration;
s8 adding chitosan oligosaccharide, anserine and brown algae oligosaccharide into the liquor, controlling the chitosan oligosaccharide content to be 0.5g/L, anserine content to be 2g/L and brown algae oligosaccharide content to be 3g/L, stirring for dissolving, and storing at-1 deg.C.
Example 2: preparation of Dipiaoling beer
S1, respectively removing seed coats and germs of clean 60 parts by mass of barley malt, 40 parts by mass of wheat malt and 10 parts by mass of burnt malt, adding water to moisten wheat for 10min, and then crushing the raw materials;
s2, preheating water in a saccharifying tank to 38 ℃, putting barley malt flour, wheat malt flour and burnt malt flour into the saccharifying tank, uniformly stirring, standing for 20min, heating to 53 ℃ at the speed of 1 ℃/min under the stirring condition, standing for 60min, heating to 67 ℃ at the speed of 1 ℃/min under the stirring condition, standing for 60min, and heating to 78 ℃ at the speed of 1 ℃/min under the stirring condition;
s3, carrying out first filtration on the saccharified mash to obtain clear wort;
s4 boiling the wort, adding 0.00025 parts by mass of bitter hops, adding 0.00018 parts by mass of fragrant hops after 45min, and stopping heating after 50 min;
s5, carrying out second filtration on the boiled wort while the wort is hot, removing filter residues, and cooling to 10.5 ℃;
s6, oxygenating the cooled wort, feeding the cooled wort into a fermentation tank, adding 0.00025 part by mass of yeast, fermenting under the conditions of 0.02 MPa-0.03 MPa and 10.5 ℃ until the sugar degree is 5.5 ℃, heating to 12.5 ℃, continuing to ferment to 4.5 ℃, boosting to 0.12MPa, keeping for 6 days, cooling to 5.5 ℃, keeping for 1 day, cooling to 0 ℃ and curing for 5 days;
s7, performing third filtration after the wine liquid is mature, removing yeast, heating to 25 ℃, then adding 0.05 wt% of chitosan-based graphene carbon spheres, adsorbing for 1h, and performing fourth filtration;
s8 adding chitosan oligosaccharide, anserine and brown algae oligosaccharide into the liquor, controlling the chitosan oligosaccharide content to be 0.01g/L, the anserine content to be 0.01g/L and the brown algae oligosaccharide content to be 0.01g/L, stirring for dissolving, and storing at 0 deg.C.
Example 3: preparation of Dipiaoling beer
S1, respectively removing seed coats and germs of 58 clean barley malt, 30 clean wheat malt and 8 clean burnt wheat malt by mass, adding water to moisten the wheat for 8min, and then crushing the raw materials;
s2, preheating water in a saccharifying tank to 37 ℃, putting barley malt flour, wheat malt flour and burnt malt flour into the saccharifying tank, uniformly stirring, standing for 18min, heating to 52 ℃ at the speed of 1 ℃/min under the stirring condition, standing for 60min, heating to 66 ℃ at the speed of 1 ℃/min under the stirring condition, standing for 60min, and heating to 77 ℃ at the speed of 1 ℃/min under the stirring condition;
s3, carrying out first filtration on the saccharified mash to obtain clear wort;
s4 boiling the wort, adding 0.00022 parts by mass of bitter hops, adding 0.00016 part by mass of fragrant hops after 45min, and stopping heating after 50 min;
s5, carrying out second filtration on the boiled wort while the wort is hot, removing filter residues, and cooling to 10.5 ℃;
s6, oxygenating the cooled wort, feeding the cooled wort into a fermentation tank, adding 0.00023 parts by mass of yeast, fermenting under the conditions of 0.025MPa and 10.5 ℃ until the sugar degree is 5.5 ℃, heating to 12.5 ℃, continuing to ferment to 4 ℃, boosting to 0.11MPa, keeping for 5 days, cooling to 5 ℃, keeping for 1 day, cooling to 0 ℃ and curing for 4 days;
s7, performing third filtration after the wine liquid is mature, removing yeast, heating to 23 ℃, then adding 0.04 wt% of chitosan-based graphene carbon spheres, adsorbing for 1h, and performing fourth filtration;
s8 adding chitosan oligosaccharide, anserine and brown algae oligosaccharide into the liquor, controlling the chitosan oligosaccharide content to be 1g/L, the anserine content to be 5g/L and the brown algae oligosaccharide content to be 5g/L, stirring for dissolving, and storing at 0 deg.C.
Example 4: preparation of Dipiaoling beer
S1, respectively removing seed coats and germs of 55 parts by mass of clean barley malt, 25 parts by mass of wheat malt and 5 parts by mass of burnt malt, adding water to moisten wheat for 5min, and then crushing the raw materials;
s2, preheating water in a saccharifying tank to 36 ℃, putting barley malt flour, wheat malt flour and burnt malt flour into the saccharifying tank, uniformly stirring, standing for 15min, heating to 52 ℃ at the speed of 1 ℃/min under the stirring condition, standing for 55min, heating to 66 ℃ at the speed of 1 ℃/min under the stirring condition, standing for 55min, and heating to 77 ℃ at the speed of 1 ℃/min under the stirring condition;
s3, carrying out first filtration on the saccharified mash to obtain clear wort;
s4 heating and boiling the wort, adding 0.00020 parts by mass of bitter hops, adding 0.00015 parts by mass of fragrant hops after 42min, and stopping heating after 48-50 min;
s5, carrying out second filtration on the boiled wort while the wort is hot, removing filter residues, and cooling to 10-10.5 ℃;
s6, oxygenating the cooled wort, feeding the cooled wort into a fermentation tank, adding 0.00022 parts by mass of yeast, fermenting under the conditions of 0.02Mpa and 10 ℃ until the sugar degree is 5 ℃, heating to 12 ℃, continuing to ferment to 4 ℃, boosting to 0.11MPa, keeping for 5 days, cooling to 5 ℃, keeping for 1 day, cooling to-1 ℃ and curing for 4 days;
s7, performing third filtration after the wine liquid is mature, removing yeast, heating to 20 ℃, then adding 0.02 wt% of chitosan-based graphene carbon spheres, adsorbing for 1h, and performing fourth filtration;
s8 adding chitosan oligosaccharide (molecular weight is 2000-3000 Da), anserine (molecular weight is less than 1000Da) and brown algae oligosaccharide (molecular weight is 2000-3000 Da) into the liquor, controlling the content of chitosan oligosaccharide in the liquor to be 0.05g/L, the content of anserine to be 2g/L and the content of brown algae oligosaccharide to be 3g/L, stirring for dissolving, and storing at-1 ℃.
The purine content of the tequila beer prepared in examples 1 to 4 was measured, and the results are shown in table 1.
TABLE 1 determination of the content (mg/L) of the different examples Dipiaoling beer
Example 1 Example 2 Example 3 Example 4
Guanine and its preparing process 5.6 4.8 4.6 5.1
Adenine 5.7 5.0 5.2 5.9
Xanthine 3.5 2.7 2.4 1.8
Hypoxanthine 3.9 2.3 3.0 3.1
Total purines 18.7 14.8 15.2 15.9
As shown in Table 1, the purine content of the beer prepared by the preparation method provided by the invention can be controlled below 20mg/L in the beer prepared by the embodiments 1-4, compared with the purine content of domestic common beer of 38-151 mg/L, the purine content of the beer is obviously reduced, and meanwhile, when the addition amount of chitosan-based graphene carbon spheres is increased, the purine content of the beer can be further reduced, and if the addition amount is 0.05 wt%, the purine content of the beer prepared by the embodiment 2 can be below 15 mg/L; compared with the embodiment 1, the content of beer in the embodiment 4 is also reduced, the difference between the two embodiments is that the molecular weight of the marine active substance added in the embodiment 4 is limited, wherein the biological activity and the physical property of the chitosan oligosaccharide and the brown algae oligosaccharide are obviously related to the molecular weight of the chitosan oligosaccharide and the brown algae oligosaccharide, the molecular weight of the chitosan oligosaccharide and the brown algae oligosaccharide is limited to 2000-3000 Da, the molecular weight of the chitosan oligosaccharide and the brown algae oligosaccharide is increased compared with that of the chitosan oligosaccharide and brown algae oligosaccharide prepared conventionally, the molecular chain length, the active amino group and the hydroxyl group are more, and the chitosan oligosaccharide and brown algae oligosaccharide have a.
Uric acid reducing animal experiment
Animal experiments were carried out on the teflons beer prepared in example 1 and example 4, and the specific experimental conditions were as follows:
after the rats were acclimatized for 7 days, the rats were weighed and randomly divided into six groups, 10 per group, namely a normal control group, a model control group, an example 1 group, an example 4 group, a comparative example group (plain beer) and a positive control group (benzbromarone). Except for the normal control group fed with the ordinary feed in the experimental process, the other 5 groups are fed with 10% yeast powder feed, 100mg/kg adenine is given in the morning for intragastric administration, 100 mg/(kg. d) oteracil potassium is injected by abdominal subcutaneous injection for 2 times, and the model is continuously built for 14 d. After the model is built, rats in each treatment group are respectively and simultaneously administrated with 2mL/100g of beer in the group 1 in the example, 2mL/100g of beer in the group 4 in the comparative group in the example, 2mL/100g of beer in the comparative group, 20mg/kg of benzbromarone in the positive control group, and 14d of distilled water for continuous administration in the normal control group and the model control group.
TABLE 1 Effect of Dipiaoling beer on serum uric acid, creatinine and urea in hyperuricemia rats
Figure BDA0002726580060000101
Note: p <0.01 compared to normal control group; compared with the model control group, the delta P is less than 0.05, and the delta P is less than 0.01
As can be seen from the results in Table 1, the serum uric acid, creatinine and urea levels in the model control group are obviously increased (P <0.01) compared with those in the normal control group, which indicates that the rat model with hyperuricemia is successfully modeled. After the rats are continuously filled with 14d, the serum uric acid, creatinine and urea of the rats in the comparative example group (the ordinary beer) are even higher than those in the model group, which indicates that the symptoms of hyperuricemia are aggravated indeed by long-term drinking of the ordinary beer; in example 1 and 4 groups of rats, serum uric acid, creatinine and urea are obviously reduced compared with a model control group, which shows that the Dipiaoling beer can inhibit the increase of serum uric acid, creatinine and urea in the serum of the rats, has good protection effect on the kidneys of the rats and improves the symptoms of hyperuricemia rats. Meanwhile, the experimental data show that the uric acid, creatinine and urea of the rats in the group of example 4 are reduced compared with those in the group of example 1, and the effect of limiting chitosan oligosaccharide (molecular weight of 2000-3000 Da), marine fish oligopeptide (goose muscle peptide molecular weight of less than 1000Da) and brown alginate oligosaccharide (molecular weight of 2000-3000 Da) in improving the uric acid reduction effect of the Dipiaoling beer is improved.
Example 5: preparation of Dipiaoling beer
S1, respectively removing seed coats and germs of 55 parts by mass of clean barley malt, 25 parts by mass of wheat malt and 5 parts by mass of burnt malt, adding water to moisten wheat for 5min, and then crushing the raw materials;
s2, preheating water in a saccharifying tank to 36 ℃, putting barley malt flour, wheat malt flour and burnt malt flour into the saccharifying tank, uniformly stirring, standing for 15min, heating to 52 ℃ at the speed of 1 ℃/min under the stirring condition, standing for 55min, heating to 66 ℃ at the speed of 1 ℃/min under the stirring condition, standing for 55min, and heating to 77 ℃ at the speed of 1 ℃/min under the stirring condition;
s3, carrying out first filtration on the saccharified mash to obtain clear wort;
s4 heating and boiling the wort, adding 0.00020 parts by mass of bitter hops, adding 0.00015 parts by mass of fragrant hops after 42min, and stopping heating after 48-50 min;
s5, carrying out second filtration on the boiled wort while the wort is hot, removing filter residues, and cooling to 10-10.5 ℃;
s6, oxygenating the cooled wort, feeding the cooled wort into a fermentation tank, adding 0.00022 parts by mass of yeast, fermenting under the conditions of 0.02Mpa and 10 ℃ until the sugar degree is 5 ℃, heating to 12 ℃, continuing to ferment to 4 ℃, boosting to 0.11MPa, keeping for 5 days, cooling to 5 ℃, keeping for 1 day, cooling to-1 ℃ and curing for 4 days;
s7, performing third filtration after the wine liquid is mature, removing yeast, heating to 20 ℃, then adding 0.02 wt% of chitosan-based graphene carbon spheres, adsorbing for 1h, and performing fourth filtration;
s8 adding chitosan oligosaccharide into the liquor, controlling the content of chitosan oligosaccharide in the liquor to be 0.5g/L, stirring to dissolve, and storing at-1 deg.C.
Example 6: preparation of Dipiaoling beer
S1, respectively removing seed coats and germs of 55 parts by mass of clean barley malt, 25 parts by mass of wheat malt and 5 parts by mass of burnt malt, adding water to moisten wheat for 5min, and then crushing the raw materials;
s2, preheating water in a saccharifying tank to 36 ℃, putting barley malt flour, wheat malt flour and burnt malt flour into the saccharifying tank, uniformly stirring, standing for 15min, heating to 52 ℃ at the speed of 1 ℃/min under the stirring condition, standing for 55min, heating to 66 ℃ at the speed of 1 ℃/min under the stirring condition, standing for 55min, and heating to 77 ℃ at the speed of 1 ℃/min under the stirring condition;
s3, carrying out first filtration on the saccharified mash to obtain clear wort;
s4 heating and boiling the wort, adding 0.00020 parts by mass of bitter hops, adding 0.00015 parts by mass of fragrant hops after 42min, and stopping heating after 48-50 min;
s5, carrying out second filtration on the boiled wort while the wort is hot, removing filter residues, and cooling to 10-10.5 ℃;
s6, oxygenating the cooled wort, feeding the cooled wort into a fermentation tank, adding 0.00022 parts by mass of yeast, fermenting under the conditions of 0.02Mpa and 10 ℃ until the sugar degree is 5 ℃, heating to 12 ℃, continuing to ferment to 4 ℃, boosting to 0.11MPa, keeping for 5 days, cooling to 5 ℃, keeping for 1 day, cooling to-1 ℃ and curing for 4 days;
s7, performing third filtration after the wine liquid is mature, removing yeast, heating to 20 ℃, then adding 0.02 wt% of chitosan-based graphene carbon spheres, adsorbing for 1h, and performing fourth filtration;
s8 adding anserine into the liquor, controlling the content of anserine in the liquor to be 2g/L, stirring for dissolving, and storing at-1 deg.C.
Example 7: preparation of Dipiaoling beer
S1, respectively removing seed coats and germs of 55 parts by mass of clean barley malt, 25 parts by mass of wheat malt and 5 parts by mass of burnt malt, adding water to moisten wheat for 5min, and then crushing the raw materials;
s2, preheating water in a saccharifying tank to 36 ℃, putting barley malt flour, wheat malt flour and burnt malt flour into the saccharifying tank, uniformly stirring, standing for 15min, heating to 52 ℃ at the speed of 1 ℃/min under the stirring condition, standing for 55min, heating to 66 ℃ at the speed of 1 ℃/min under the stirring condition, standing for 55min, and heating to 77 ℃ at the speed of 1 ℃/min under the stirring condition;
s3, carrying out first filtration on the saccharified mash to obtain clear wort;
s4 heating and boiling the wort, adding 0.00020 parts by mass of bitter hops, adding 0.00015 parts by mass of fragrant hops after 42min, and stopping heating after 48-50 min;
s5, carrying out second filtration on the boiled wort while the wort is hot, removing filter residues, and cooling to 10-10.5 ℃;
s6, oxygenating the cooled wort, feeding the cooled wort into a fermentation tank, adding 0.00022 parts by mass of yeast, fermenting under the conditions of 0.02Mpa and 10 ℃ until the sugar degree is 5 ℃, heating to 12 ℃, continuing to ferment to 4 ℃, boosting to 0.11MPa, keeping for 5 days, cooling to 5 ℃, keeping for 1 day, cooling to-1 ℃ and curing for 4 days;
s7, performing third filtration after the wine liquid is mature, removing yeast, heating to 20 ℃, then adding 0.02 wt% of chitosan-based graphene carbon spheres, adsorbing for 1h, and performing fourth filtration;
s8 adding alginate-derived oligosaccharide into the liquor, controlling the content of alginate-derived oligosaccharide in the liquor to be 3g/L, stirring for dissolving, and storing at-1 deg.C.
The above embodiments are the best mode for carrying out the present invention, but the embodiments of the present invention are not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be regarded as equivalent substitutions and are included in the scope of the present invention.

Claims (10)

1. A low purine beer with uric acid reducing effect is characterized by comprising at least one of chitosan oligosaccharide, marine fish oligopeptide and brown algae oligosaccharide.
2. The low purine beer having uric acid lowering efficacy according to claim 1, wherein: purine content <20 mg/L.
3. The low purine beer having uric acid lowering efficacy according to claim 1, wherein: the content of the chitosan oligosaccharide in the beer is 0-1 g/L; the content of the marine fish oligopeptide in the beer is 0-5 g/L; the content of the brown algae oligosaccharide in the beer is 0-5 g/L.
4. The low purine beer having uric acid lowering efficacy according to claim 1, wherein: the molecular weight of the chitosan oligosaccharide is 300-3000 Da.
5. The low purine beer having uric acid lowering efficacy according to claim 1, wherein: the marine fish oligopeptide comprises dipeptide and tripeptide, and the molecular weight of the marine fish oligopeptide is less than 1000 Da.
6. The low purine beer having uric acid lowering efficacy according to claim 1, wherein: the brown algae oligosaccharide is extracted from kelp, undaria pinnatifida, sargassum fusiforme and sargassum horneri; the molecular weight of the brown algae oligosaccharide is 500-3000 Da.
7. The method for preparing low-purine beer with uric acid reducing effect according to any one of claims 1 to 6, comprising the following steps:
s1, respectively removing seed coats and germs of clean 55-60 parts by mass of barley malt, 25-40 parts by mass of wheat malt and 5-10 parts by mass of burnt malt, adding water to moisten wheat for 5-10 min, and then crushing;
s2, preheating water in a saccharifying tank to 36-38 ℃, then putting the barley malt powder, the wheat malt powder and the burnt wheat malt powder which are obtained by crushing in S1 into the saccharifying tank, uniformly stirring, standing for 15-20 min, then heating to 52-53 ℃ at the speed of 1 ℃/min under the stirring condition, standing for 55-60 min, heating to 66-67 ℃ at the speed of 1 ℃/min under the stirring condition, standing for 55-60 min, and heating to 77-78 ℃ at the speed of 1 ℃/min under the stirring condition;
s3, carrying out first filtration on the saccharified mash to obtain clear wort;
s4 heating and boiling the wort, adding 0.00020-0.00025 parts by mass of bitter hops, adding 0.00015-0.00018 parts by mass of fragrant hops after 42-45 min, and stopping heating after 48-50 min;
s5, carrying out second filtration on the boiled wort while the wort is hot, removing filter residues, and cooling to 10-10.5 ℃;
s6, oxygenating the cooled wort, feeding the cooled wort into a fermentation tank, adding 0.00022-0.00025 parts by mass of yeast, fermenting under the conditions of 0.02 Mpa-0.03 MPa and 10-10.5 ℃ until the sugar degree is 5-5.5 ℃, heating to 12-12.5 ℃, continuing to ferment to 4-4.5 ℃, boosting to 0.11-0.12 MPa, keeping for 5-6 days, cooling to 5-5.5 ℃, keeping for 1 day, cooling to-1-0 ℃ and curing for 4-5 days;
s7, performing third filtration after the wine liquid is mature, removing yeast, heating to 20-25 ℃, then adding chitosan-based graphene carbon spheres, adsorbing for 1h, and performing fourth filtration;
s8 adding chitosan oligosaccharide, marine fish oligopeptide and brown algae oligosaccharide into the liquor, stirring to dissolve, and storing at-1-0 ℃.
8. The method for preparing low purine beer with uric acid reducing effect according to claim 7, wherein: in the step S2, the material-liquid ratio of the total mass of the barley malt flour, the wheat malt flour and the burnt malt flour to water is 4: 1-5: 1.
9. The method for preparing low purine beer with uric acid reducing effect according to claim 7, wherein: in the step S7, the addition amount of the chitosan-based graphene carbon spheres is 0.02-0.05 wt% of the wine.
10. The method for preparing low purine beer with uric acid reducing efficacy according to claim 9, wherein the chitosan-based graphene carbon beads are prepared by the following steps:
dissolving chitosan in 1.5-2% acetic acid aqueous solution, continuously adding graphene powder, stirring uniformly, slowly dripping chitosan-graphene suspension into 2-3 mol/L NaOH solution to prepare balls, filtering out solid pellets after 2-3 h, washing with water to be neutral, adding the solid pellets into 25-30% glutaraldehyde solution for crosslinking, taking out after 3h of crosslinking, drying, and carbonizing the dried pellets at 400-420 ℃ for 0.5-0.8 h under the condition of inert atmosphere; and cooling to room temperature, washing away impurities with water, and drying to obtain the chitosan-based graphene carbon spheres.
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