CN104099312B - A kind of liquid compound enzyme for brewing and preparation method thereof - Google Patents

A kind of liquid compound enzyme for brewing and preparation method thereof Download PDF

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Publication number
CN104099312B
CN104099312B CN201410344967.1A CN201410344967A CN104099312B CN 104099312 B CN104099312 B CN 104099312B CN 201410344967 A CN201410344967 A CN 201410344967A CN 104099312 B CN104099312 B CN 104099312B
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enzyme
liquid
liquid compound
brewing
preparation
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CN104099312A (en
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李洪兵
朱永明
杨志祥
高志忠
蔡先红
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HUNAN XINHONGYING BIO-ENGINEERING Co Ltd
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HUNAN XINHONGYING BIO-ENGINEERING Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/96—Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12C—BEER; PREPARATION OF BEER BY FERMENTATION; PREPARATION OF MALT FOR MAKING BEER; PREPARATION OF HOPS FOR MAKING BEER
    • C12C5/00—Other raw materials for the preparation of beer
    • C12C5/004—Enzymes
    • C—CHEMISTRY; METALLURGY
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    • C12C—BEER; PREPARATION OF BEER BY FERMENTATION; PREPARATION OF MALT FOR MAKING BEER; PREPARATION OF HOPS FOR MAKING BEER
    • C12C7/00—Preparation of wort
    • C12C7/04—Preparation or treatment of the mash
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    • C12N9/2414—Alpha-amylase (3.2.1.1.)
    • C12N9/2417—Alpha-amylase (3.2.1.1.) from microbiological source
    • C12N9/242—Fungal source
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    • C12N9/2425—Beta-amylase (3.2.1.2)
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    • C12Y302/01041—Pullulanase (3.2.1.41)
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C12Y302/01068—Isoamylase (3.2.1.68)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C12Y302/01073—Licheninase (3.2.1.73)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C12Y302/01108—Lactase (3.2.1.108)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
    • C12Y304/22—Cysteine endopeptidases (3.4.22)
    • C12Y304/22002—Papain (3.4.22.2)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
    • C12Y304/22—Cysteine endopeptidases (3.4.22)
    • C12Y304/22003—Ficain (3.4.22.3)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y401/00—Carbon-carbon lyases (4.1)
    • C12Y401/01—Carboxy-lyases (4.1.1)
    • C12Y401/01005—Acetolactate decarboxylase (4.1.1.5)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y503/00—Intramolecular oxidoreductases (5.3)
    • C12Y503/01—Intramolecular oxidoreductases (5.3) interconverting aldoses and ketoses (5.3.1)
    • C12Y503/01018—Glucose isomerase (5.3.1.18)

Abstract

The invention discloses a kind of liquid compound enzyme for brewing and preparation method thereof, belong to liquid compound enzyme preparing technical field, with brewing liquid enzyme formulation as primary raw material, science compound sugar, polyalcohol, amino acid, thickener, albumin, the hop extraction thing that the stabilizers such as metal ion are more stable with promoting enzyme preparation, extracts of Chinese herbal medicine, antibacterial peptides etc. have the material of antisepsis, instead of the interpolation of preservative completely, the most effectively inhibit harmful microbe breeding and growth, and the activity of stable liquid compound enzyme can be worked in coordination with other component, fundamentally improve biological stability and the enzyme activity stability of liquid compound enzyme, effectively extend the shelf-life of liquid enzyme formulation.The liquid compound enzyme of preparation is 168-214CFU/mL in 12 months total plate counts of room temperature storage；Storing 12 months under the conditions of 40 DEG C and 0 DEG C, enzyme loss late alive is respectively 2-2.5% and 1-1.7%.

Description

A kind of liquid compound enzyme for brewing and preparation method thereof
Technical field
The present invention relates to liquid compound enzyme, be specifically related to a kind of liquid compound enzyme for brewing and preparation side thereof Method.
Background technology
Liquid enzyme formulation is a kind of easy to use, environmental protection, the raw material that biological degradability is good.Single-minded catalysis characteristics And the reaction condition of gentleness, more and more used in industrial quarters and paid attention to.In use liquid enzyme formulation have with Lower advantage: 1) liquid enzyme formulation is dispersed in substrate with molecule or graininess, and decentralization is big, absorbs fast, can relatively rapidly send out Raw enzyme digestion reaction, enzymatic activity is high；2) it is prone to divided dose, the most compounding and secondary operations, easy to use；3) be conducive to improving enzyme system The utilization rate of agent, 4) directly use, need not dissolve and activate；5) purity is high, impurity content is low；6) production process is short, low cost； 7) do not result in dust pollution during production and processing and use, be conducive to protecting environment；But, liquid enzyme formulation volume is relatively Greatly, carrying, transport, store all inconveniences, enzymatic activity is easily subject to the storages such as high temperature, freezing, illumination, Acidity of Aikalinity, oxidation, humidity The impact of environment and significantly reduce, and because the water activity of liquid enzyme formulation is higher, it is easy to polluted by microorganism And inactivate.Therefore, the application of liquid enzyme formulation is restricted, and topmost reason is exactly its less stable, including biological steady Qualitative and physics, chemical stability.
In order to solve the problems referred to above, existing enzyme preparation manufacturer just by concentrated, separate after liquid enzyme formulation add Add stabilizer, preservative, surfactant, metal ion etc., to improve the biological stability of liquid enzyme formulation, abiotic stable Property and using effect, widen its range of application in fields such as food, weaving, papermaking, washing, feeds, thus, liquid compound enzyme Arising at the historic moment, on the whole, liquid compound enzyme is divided into two kinds, and one is single liquid compound enzyme, i.e. by a kind of liquid enzyme formulation Process with additive；Another kind is plurality of liquid compound enzyme, is to be processed by plurality of liquid enzyme preparation and additive.Cause For unicity and the diversity of action condition, the otherness of enzyme preparation substrate specificity, at present, market is used for industrialized liquid Enzyme system is based on single liquid compound enzyme, and plurality of liquid compound enzyme is less, almost without.
The compounding primary precondition of liquid compound enzyme is application, specific to the product performance of processed finished products, as Fruit do not consider the factor of product performance and carry out blindly in order to improve the compounding of stability, not only will not effectively play enzyme preparation Effect, serious side effect can be produced on the contrary, even can lead to great production accident.Therefore the preparation of liquid compound enzyme to be combined Close and consider many factors.
At present, the liquid compound enzyme on market is concentrated mainly on the technology necks such as papermaking, washing, food, feed, alcohol be standby Territory, and based on single liquid compound enzyme, the surfaces such as wherein adding stabilizer has the polyalcohols such as glycerine and xanthans, gelatin, CMC The metals such as macromolecular compound, sodium chloride such as activating agent, polyalcohols surfactant, AEO compound, pvp The preservatives etc. such as chloride, cationic and anionic surfactant, Sodium Benzoate, the kind of additive also with the kind of enzyme, same enzyme Substrate specificity relevant with many factors such as chemical property, enzyme action effects with the physics of condition, additive, not can unify Add, use.
Unlike other fermentation industry, in vigorous froth breaking campaign, brewing is from start to finish in maximum The holding foam of limit, beer holding property of bubble is one of important physics and chemistry and organoleptic indicator of beer.Beer is different from it in appearance One important feature of its beverage: exactly when pouring in cup, the foam of pure white exquisiteness can be produced, give cleaning, clearly feel, The drink promoting people is intended to.Therefore the holding property of bubble of beer becomes consumer and recognizes the key character of Beer Brand.The holding property of bubble of beer Typically refer to: the foaming characteristic of beer, persistence and tack.Some stabilizer of liquid compound enzyme and surfactant can play by force The effect of power defoamer, the interpolation of sodium chloride can be that beer produces saline taste, affects mouthfeel, and human body can be produced by the interpolation of preservative Poison ....
Therefore, for brewing industry, a kind of safe and stable, action effect significant liquid compound enzyme of preparation Restrictive condition is more, and especially seem difficulty, and the plurality of liquid compound enzyme preparing a kind of applicable brewing is more difficult, at present, Yet there are no the report of disclosed beer plurality of liquid compound enzyme.Chinese patent CN101617740B discloses a kind of enzyme retention alive High forage liquid phytase preparation.The present invention is addition Macrogol 6000, trehalose, benzoic acid in phytase original enzyme liquid Sodium, pure water, regulation pH value is 4.5～5.5: wherein in the hydroxyl in Macrogol 6000 and zymoprotein molecular surface and activity thereof The carboxyl of the heart, amino etc. combine with covalent bond, hydrogen bond and Van der Waals force etc., so that the activated centre of zymoprotein molecule and whole Zymoprotein molecule is protected；Trehalose (stabilizer), improves the osmotic pressure of enzyme solutions, makes the degree of unfolding of zymoprotein molecule drop Low, molecule is relatively in dewatering state and shrinks, thus improves stability；Sodium Benzoate (preservative) and preferably pH scope (4.5 ～5.5), all play the effect of the biological stability improving liquid enzymes.The open a kind of cyclodextrin of Chinese patent CN101717765B Glycosyltransferase compound enzyme preparation, it is characterised in that this enzyme preparation is a kind of liquid enzyme formulation；Consisting of: cyclodextrin Portugal Glucosyl transferase, glycerine, CaCl2, PEG400 and gelatin；Described cyclodextrin glycosyltransferase is that protein content is The zymotic fluid of 3g/L-5g/L, glycerol concentration is 10%-20% (volume fraction), and CaCl2 concentration is 0.1g/L-0.2g/L, PEG400 concentration is 5%-10% (volume fraction), and gelatin concentration is 0.5%-1% (mass fraction).There is extremely excellent storage Depositing stability, preserve 60 days at 40 DEG C, its enzyme retention rate alive is still up to more than 95%, when solving due to product transport or store Between the longer problem causing products application hydraulic performance decline.The liquid enzyme formulation preparation method that the present invention provides is simple, raw material sources Enrich and low cost, be particularly suitable for industrially promoting.Chinese patent CN103131557B discloses a kind of liquid detergent Use enzyme preparation compound stabilizer, relate to enzyme stabilizers, solve enzyme stability in existing liquid detergent poor, and tradition enzyme stabilizers Mainly use borax, the problem that human body is existed potential hazard.A kind of liquid detergent enzyme preparation compound stabilizer, by following Weight portion each component composition: cyclodextrin 0.05～20, calcium chloride 0.05～20, glycerine and propylene glycol mixture 1～10, non-from Sub-surface activating agent AEO 1～60, anion surfactant sodium alkyl benzene sulfonate 1～60；Wherein said Glycerine and propylene glycol mixture in glycerine: the weight ratio of propane diols is 2:6～10.The present invention can simplify the life of liquid detergent Producing and formulation procedures, significantly improve and extend the activity of enzyme preparation in liquid detergent, materials safety is nontoxic, harmless. Liquid enzyme formulation disclosed above and stabilizer are all not suitable for brewing and produce.
To sum up, safe and stable, the effect significant plurality of liquid compound enzyme of preparing a kind of applicable brewing have wide The market space and good market value.
Summary of the invention
Solved by the invention technical problem is that one or more are concentrated, separate liquid enzyme formulation according to beer make Make stabilizer and the rush such as purposes rational allocation, and science compound sugar, polyalcohol, amino acid, thickener, albumin, metal ion Enter the more stable hop extraction thing of enzyme preparation, extracts of Chinese herbal medicine, antibacterial peptide etc. and there is the material of antisepsis, effectively carry The enzyme activity stability of high liquid compound enzyme and biological stability, played important catalyst action in brewing, Further improve the quality index such as mouthfeel and foaming properties of beer simultaneously, preservative need not be added, prepare and a kind of be suitable for beer Safe and stable, the effect significant liquid compound enzyme brewageed.
The primary and foremost purpose of the present invention is to provide a kind of liquid compound enzyme for brewing.
In order to achieve the above object, the present invention is by the following technical solutions:
A kind of liquid compound enzyme for brewing, is prepared by the raw material of following parts by weight:
Liquid enzyme formulation 30-50 part, hops extract 5-15 part, sugar 5-10 part, natural 4-9 part, medium-height grass Medicament extracting solution 5-8 part, antibacterial peptide 4-6 part, thickener 3-5 part, beta-schardinger dextrin 3-5 part, polyalcohol 2-5 part, glutathione 2-4 Part, metal chloride 1-3 part, ammonium sulfate 1-2 part, cysteine 0.5-1 part, bovine serum albumin(BSA) 0.1-0.3 part.
The preference temperature of described liquid enzyme formulation is 35-115 DEG C, and optimum pH is 2.5-6；
Further, described liquid enzyme formulation selects free dextranase, zytase, middle temperature amylase, AMS, resistance to High-temperatureα-amylase, fungal alpha-amylase, isoamylase, carbohydrase, beta amylase, Pullulanase, papain, pineapple egg White enzyme, neutral proteinase, acid protease, proline protein enzyme, ficin, 1,4 beta-glucanase, heatproof beta glucan Complex enzyme, beta glucan complex enzyme, mannonase pentosanase, acid phosphatase, lipase, phosphate, glucoside Enzyme, glucose isomerase, tannase, lactase, catalase, alpha-acetolactate decarboxylase composition group in any one or Several, but it is not limited to these enzymes, it is possible to it is applicable to other industrial enzyme preparations.Liquid enzymes not only includes under natural conditions The enzyme that bacterial strain produces, and include the enzyme produced by genetic engineering modified engineering bacteria；
Preferably, described liquid enzyme formulation is by middle temperature amylase, dextranase, zytase 3-4:1-1.5 by volume: 0.5-1 uniformly mixes；
Further, the preparation method of described hops extract is: hops is put in supersonic wave cleaning machine in 200W, 40KHz cleans 10-15min, drains, and pulverizes immediately after-18-22 DEG C of freezing 1-2h, freezing thickness of feed layer 3-5cm, Grinding particle size 0.5-3mm, adds ethanol and the mixture of propyl alcohol pulverizing hops weight 1-3 times, ethanol and propyl alcohol mixing Mass ratio is 2-4:1-3, is 3.0-3.5 with breast acid for adjusting pH value, in power 150-300W, frequency 2000Hz, temperature 20-35 DEG C Under the conditions of carry out Microwave Extraction 20-30min, simultaneously at power 200-300W, carry out ultrasonic wave under the conditions of frequency 30-40KHz auxiliary Help extraction；Insulation 1-3h, then, carries out Microwave Extraction 15-20min, simultaneously under the conditions of power 200-400W, frequency 2000Hz At power 300-500W, under the conditions of frequency 40-50KHz, carry out ultrasonic assistant extraction, be finally naturally cooling to room temperature, filter, Filtrate, in electric-field intensity 35-45kV/cm, burst length 400-600 μ s, carries out high-tension pulse under the conditions of pulse frequency 200-300Hz Rush electric field (PEF) sterilization 20-30min and i.e. obtain hops extract；
Described hops is fresh mature hops, compression film clips or compressing grains hops, preferably fresh mature hops； When using pellet hop, need not clean；
Preferably, the sodium bicarbonate solution that described Ultrasonic Cleaning uses mass percent concentration to be 0.2-0.8% is clear Lotion；
Preferably, the mass ratio of described ethanol and propyl alcohol mixing is 3:2；
Preferably, described Microwave Extraction uses batch (-type) to extract, i.e. microwave irradiation 20s stops 10s, so circulates；
Described concentration of alcohol >=75%.
Further, during described sugar is trehalose, GL-B, glucose one or more；
Preferably, described sugar is that trehalose, GL-B, glucose 5-8:1-3:2-4 in mass ratio uniformly mixes.
Further, during described natural is grape pip procyanidin, Rosmarinus officinalis extract and apricot leaf extract Any or several combination；
Preferably, the mass ratio of described grape pip procyanidin, Rosmarinus officinalis extract and the mixing of apricot leaf extract is 4-6: 2-4:1-3；
Further, the preparation method of described extracts of Chinese herbal medicine is: count by weight, accurately weighs Radix Astragali 60-70 Part, Radix Angelicae Sinensis 55-65 part, Radix Codonopsis 40-45 part, Radix Glycyrrhizae 40-45 part, cordate houttuynia 35-45 part, Divine Comedy 35-45 part, honeysuckle 25-35 Part, Poria cocos 20-30 part, polygala root 15-25 part, fry fennel 15-25 part, bighead atractylodes rhizome 10-20 part, bark of official magnolia 10-20 part；Respectively by above-mentioned It is less than 2 millimeters that herbal medicine is crushed to particle diameter, then uniformly mixes and add the water of 3-6 times of weight in container, controls temperature 70 DEG C-90 DEG C keep 2-4h, are then cooled to 45-60 DEG C, and the mixing enzyme preparation adding mixed material gross weight 5-10% carries out enzyme Solve, be 5.5-6.8 with breast acid for adjusting pH value, enzymolysis 2-4h, finally adds the mixed of 0.5-3 times of w ethanol of mixed material and propyl alcohol Compound, controls temperature and keeps 3-4h to 60 DEG C-78 DEG C, filter, obtain the first filtrate；Add the water of 1-3 times of weight of filter residue, control temperature Spend 85 DEG C-95 DEG C and keep 1-3h, be then cooled to 25-35 DEG C, filter, obtain the second filtrate；First filtrate and the second filtrate are pressed Merge according to mass ratio 2-3:1-2, in electric-field intensity 35-45kV/cm, burst length 400-600 μ s, pulse frequency 300-400Hz Under the conditions of carry out high-pressure pulse electric (PEF) sterilization 20-30min i.e. obtain extracts of Chinese herbal medicine；
Preferably, the mass ratio that described first filtrate and the second filtrate merge is 2.5:1.5；
The parts by weight of described mixed enzyme consist of: endo-beta-glucanase 10-20 part, outer 1,4 beta-glucanase 10-20 part, and β- Glucuroide 10-15 part, zytase 15-20 part, pentosanase 15-20 part, Pullulanase 20-30 part, beta amylase 10- 15 parts, acid protease 10-15 part, pectase 10-15 part, seminase 10-15 part, glucose oxidase 5-10 part is acid Phosphatase 5-10 part, lipase 5-8 part；
The mass ratio of described ethanol and propyl alcohol mixing is 1:1-2, described concentration of alcohol >=75%.
Further, during described polyalcohol is polyethylene glycol, glycerine, sorbierite, dithiothreitol (DTT), thin base ethanol Plant or several；
Preferably, described polyalcohol is by polyethylene glycol, glycerine, sorbierite, dithiothreitol (DTT) 8-10:4-6 in mass ratio: 2-4:1-3 uniformly mixes.
Further, described thickener is carragheen, Arabic gum, gelatin, xanthans, algin, CMC, agar, honeybee In glue, lac, pectin, guar gum, Chinese honey locust carbohydrate gum, linseed glue, konjac glucomannan one or more；
Preferably, described thickener is by carragheen, Arabic gum, gelatin, xanthans, CMC 3-5:2-4:1-in mass ratio 3:1-2:0.5-1 uniformly mixes.
Further, one during described metal chloride is zinc chloride, calcium chloride, sodium chloride, magnesium chloride, iron chloride or Multiple；
Preferably, described metal chloride is zinc chloride, calcium chloride, sodium chloride, magnesium chloride, iron chloride 3-5 in mass ratio: 2-3:1-3:1-2:0.1-0.3 uniformly mixes.
Another object of the present invention is to provide the preparation method of a kind of liquid compound enzyme being applied to brewing, including as follows Step:
In terms of parts by weight, the most accurately weigh thickener according to above-mentioned formula, add the sterilized water of its quality 4-6 times, Soaking at room temperature 3-8h, the stirring of intensification limit, limit, 80-90 DEG C of insulation 20-30min, stirring, fully dissolve, continue insulation, while stirring It is sequentially added into beta-schardinger dextrin, polyalcohol, sugar, metal chloride, is then cooled to 25-35 DEG C, be sequentially added into sulfuric acid while stirring Ammonium, cysteine, glutathione, antibacterial peptide, natural, bovine serum albumin(BSA), extracts of Chinese herbal medicine, hops are extracted Liquid, liquid enzyme formulation fully dissolve, mix, and are 2.5-6 by lactic acid regulation liquid compound enzyme pH value, stablize 0.5-2h, while in Electric-field intensity 35-45kV/cm, burst length 400-600 μ s, carry out high-pressure pulse electric under the conditions of pulse frequency 300-400Hz (PEF) sterilization, aseptic filtration, sterile filling i.e. obtain liquid compound enzyme.
A further object of the invention is the application in brewing of the liquid compound enzyme.
Using method:
Beer liquid compound enzyme of the present invention is added brew kettle by the saccharification stage
Addition: according to the difference of malt quality and the ratio height of brewageing auxiliary material, every t dried malt adds 500- 1000ml；
Action condition: optimum PH 4.5-5.5；Optimum temperature 35-68 DEG C；Optimal material water quality compares 1:4.5-5.5.
Beneficial effect:
1. hops extract, the extracts of Chinese herbal medicine that prepared by the present invention use low temperature extractive technique, to greatest extent will Active ingredient in hops and Chinese herbal medicine retains, and compounds with antibacterial peptide science, instead of the interpolation of preservative completely, does not only has Effect inhibit harmful microbe breeding and growth, and the activity of stable liquid compound enzyme can be worked in coordination with other component, Fundamentally improve biological stability and the enzyme activity stability of liquid compound enzyme, effectively extend guaranteeing the quality of liquid enzyme formulation Phase.The liquid compound enzyme of preparation is 168-214CFU/mL in 12 months total plate counts of room temperature storage；Under the conditions of 40 DEG C and 0 DEG C Storing 12 months, enzyme loss late alive is respectively 2-2.5% and 1-1.7%.
2. the present invention is in long experimental basis, and science has compounded the polyalcohol with defoamer effect, polyalcohol It is a class effect preferable enzyme preparation stabilizer, with other component such as hops extract, sugar, thickener, beta-schardinger dextrin, medium-height grass Medicament extracting solutions etc. together, the most do not produce impact to saccharification wort, zymotic fluid, the foaming properties of finished product and mouthfeel, have on the contrary Promoted, achieved beyond thought effect.Compared with commercial liquid carbohydrase, malt flavor is obvious with hops fragrance, soft Coordinating, foaming properties is excellent, and bubble the holding property time is long；Turbidity is relatively low, and diacetyl content is relatively low, and fermentation is good, and alcoholic strength is higher, becomes Savor beer either organoleptic indicator or physical and chemical index has precedence over the most far away ordinary beer compound enzyme, met or exceeded national existing Row beer top grade standard.
3. natural and glutathione science are compounded in liquid enzyme formulation by the present invention, have not only acted as stronger Antioxidation, it is to avoid the dissolved oxygen in liquid enzyme formulation is to enzyme molecule protein and the oxidation of other component, prior It is the nitration reaction and the non-enzymatic glycation that effectively inhibit enzyme preparation albumen of the OPC in natural, aobvious Write the vigor stability that improve liquid enzyme formulation albumen.
4. the bovine serum albumin(BSA) in the present invention is a kind of globulin in cow's serum, comprises 583 amino acid residues, can Effectively prevent decomposition and the non-specific adsorption of enzyme, more significantly to some restriction endonuclease effect, with other component such as: gluathione The science such as peptide, metal chloride, polyalcohol, ammonium sulfate compound, and substantially increase the enzyme stability alive of liquid compound enzyme.
5. the preparation method technique of the liquid compound enzyme of the present invention is simple and convenient to operate, can produce by large-scale industrial, uses High-pressure pulse electric cold sterilization technology (PEF), both ensure that the biochemical activity of liquid enzyme formulation albumen, and had the most effectively killed and press down Make breeding and the growth of microorganism, significantly improved enzyme stability alive and the biological stability of liquid enzyme formulation, effectively extend Shelf-life of liquid enzyme formulation.
Detailed description of the invention
The present invention is described below by specific embodiment.Unless stated otherwise, technological means used in the present invention It is method known in those skilled in the art.It addition, embodiment is interpreted as illustrative, and the unrestricted present invention Scope, the spirit and scope of the invention are limited only by the claims that follow.To those skilled in the art, without departing substantially from this On the premise of invention spirit and scope, the various changes carrying out the material component in these embodiments and consumption or change are also Belong to protection scope of the present invention.
Following primary raw material is originated:
The most separated, the liquid middle temperature amylase of concentration, dextranase, zytase, carbohydrase are given birth to by Hunan letter Thing Science and Technology Co., Ltd. produces, wherein:
The diastatic vigor of middle temperature is 500-2000U/mL, and optimal reactive temperature is 60-70 DEG C, and optimum pH is 5-6；
The vigor of dextranase is 5000-12000U/mL, and optimal reactive temperature is 40-65 DEG C, and optimum pH is 4-5.5；
The vigor of zytase is 5000-20000U/mL, and optimal reactive temperature is 50-65 DEG C, and optimum pH is 3.5-6；
The vigor of carbohydrase is 150000-200000U/mL, and optimal reactive temperature is 60-70 DEG C, and optimum pH is 5-6.
2. fresh mature hops picks up from Xinjiang hop cultivars limited company planting base.
3. sugar, thickener, beta-schardinger dextrin, cysteine are purchased from Quan Wang bio tech ltd, Shanghai.
4. grape pip procyanidin is bought in Earthquake of Anyang station in Henan Jing Sen bio tech ltd, active ingredient: OPC/ Polyphenol, Assay OPC95%/polyphenol90%；
Ginkgo biloba p.e is bought in Xuzhou Heng Kai ginkgo product Co., Ltd, and 2010 editions ginkgo leaves of Chinese Pharmacopoeia extract Thing, total flavonoids >=24%, equipment registration trade mark: Gongsun hall Ginkgo Town；
Rosmarinus officinalis extract buys the gloomy source book on Chinese herbal medicine natural products limited company in Henan, main component: Rosmarinic acid, Rosmanol etc., content 85%, registration mark gloomy source book on Chinese herbal medicine；
The said goods is pressed powder.
5. antibacterial peptide is purchased from Guangdong Tianhao Biological Engineering Co., Ltd., for food antibacterial peptide K-100, model K- 100。
6. glutathione is purchased from Rong Sheng bio tech ltd, Xi'an, for food glutathione, glutathione content 99%, production code member: P0200375.
7. bovine serum albumin(BSA) is purchased from Shanghai Ru Ji biotechnology Development Co., Ltd, the domestic cas:9048-46-8 of 1g.
The preparation of embodiment 1 hops extract
The preparation method of described hops extract is: fresh mature hops is put in supersonic wave cleaning machine in 200W, 40KHz cleans 12min, drains, and pulverizes immediately after-20 DEG C of freezing 1.5h, freezing thickness of feed layer 4cm, grinding particle size 2mm, adds ethanol and the mixture of propyl alcohol pulverizing hops weight 2 times, and the mass ratio of ethanol and propyl alcohol mixing is 3:2, uses Breast acid for adjusting pH value is 3.2, carries out microwave batch (-type) extraction under the conditions of power 200W, frequency 2000Hz, temperature 30 DEG C 25min, simultaneously at power 300W, carries out ultrasonic assistant extraction under the conditions of frequency 35KHz；Insulation 2h, then, at power Carry out Microwave Extraction 18min under the conditions of 300W, frequency 2000Hz, simultaneously at power 400W, carry out ultrasonic under the conditions of frequency 45KHz Ripple assisted extraction, is finally naturally cooling to room temperature, filters, and filtrate is in electric-field intensity 40kV/cm, burst length 500 μ s, pulse frequency Carry out high-pressure pulse electric (PEF) sterilization 25min under the conditions of rate 250Hz and i.e. obtain hops extract；
The sodium bicarbonate solution that described Ultrasonic Cleaning uses mass percent concentration to be 0.2-0.8% is cleaning agent；
Described batch (-type) is extracted as microwave irradiation 20s, stops 10s, so circulates；
Described concentration of alcohol is 80%.
The preparation of embodiment 2 extracts of Chinese herbal medicine
The preparation method of described extracts of Chinese herbal medicine is: count by weight, accurately weighs the Radix Astragali 65 parts, Radix Angelicae Sinensis 60 parts, Radix Codonopsis 42 parts, 42 parts of Radix Glycyrrhizae, cordate houttuynia 40 parts, Divine Comedy 40 parts, honeysuckle 30 parts, 25 parts of Poria cocos, polygala root 20 parts, fry fennel 20 Part, the bighead atractylodes rhizome 15 parts, the bark of official magnolia 15 parts；It is less than 2 millimeters that said herbal medicine is crushed to particle diameter respectively, then uniformly mixes in container Merge the water adding 5 times of weight, control temperature 80 DEG C and keep 3h, be then cooled to 50 DEG C, add mixed material gross weight 8% Mixing enzyme preparation carries out enzymolysis, with breast acid for adjusting pH value be 6.2, enzymolysis 3h, finally add 1.8 times of w ethanol of mixed material and The mixture of propyl alcohol, the mass ratio of ethanol and propyl alcohol mixing is 1:2, controls temperature and keeps 4h to 70 DEG C, filters, obtain the first filter Liquid；Add the water of 2 times of weight of filter residue, control temperature 90 DEG C and keep 2h, be then cooled to 30 DEG C, filter, obtain the second filtrate；By One filtrate and the second filtrate merge according to mass ratio 2.5:1.5, in electric-field intensity 40kV/cm, burst length 500 μ s, pulse frequency Carry out high-pressure pulse electric (PEF) sterilization 25min under the conditions of rate 350Hz and i.e. obtain extracts of Chinese herbal medicine；
The parts by weight of described mixed enzyme consist of: endo-beta-glucanase 20 parts, outer 1,4 beta-glucanase 18 parts, β-glucose Glycosides enzyme 12 parts, zytase 18 parts, pentosanase 18 parts, Pullulanase 25 parts, beta amylase 12 parts, acid protease 12 parts, Pectase 12 parts, seminase 12 parts, glucose oxidase 6 parts, acid phosphatase 8 parts, 6 parts of lipase；
Described concentration of alcohol is 75%.
Embodiment 3
A kind of liquid compound enzyme for brewing, is prepared by the raw material of following parts by weight:
Liquid enzyme formulation 40 parts, hops extract 10 parts, sugar 8 parts, natural 7 parts, extracts of Chinese herbal medicine 6 Part, antibacterial peptide 5 parts, thickener 4 parts, beta-schardinger dextrin 4 parts, polyalcohol 3 parts, glutathione 3 parts, metal chloride 2 parts, ammonium sulfate 2 parts, cysteine 0.8 part, bovine serum albumin(BSA) 0.2 part.
Described liquid enzyme formulation is uniformly mixed by middle temperature amylase, dextranase, zytase 3.5:1.2:0.8 by volume Close；
Described sugar is that trehalose, GL-B, glucose 7:2:3 in mass ratio uniformly mixes；
Described natural is by grape pip procyanidin, Rosmarinus officinalis extract and apricot leaf extract 5:3 in mass ratio: 2 uniformly mix；
Described thickener is uniform by carragheen, Arabic gum, gelatin, xanthans, CMC 4:3:2:1.5:0.8 in mass ratio Mixing；
Described polyalcohol is uniformly mixed by polyethylene glycol, glycerine, sorbierite, dithiothreitol (DTT) 9:5:3:2 in mass ratio；
Described metal chloride is zinc chloride, calcium chloride, sodium chloride, magnesium chloride, iron chloride 4:2.5:2 in mass ratio: 1.5:0.2 uniformly mixes；
Described hops extract is prepared by embodiment 1；
Described extracts of Chinese herbal medicine is prepared by embodiment 2；
Preparation method: in terms of parts by weight, the most accurately weighs thickener according to above-mentioned formula, adds its quality 5 times Sterilized water, soaking at room temperature 6h, the stirring of intensification limit, limit, 85 DEG C of insulation 25min, stirring, fully dissolve, continue insulation, while stirring Be sequentially added into beta-schardinger dextrin, polyalcohol, sugar, metal chloride, be then cooled to 30 DEG C, be sequentially added into while stirring ammonium sulfate, Cysteine, glutathione, antibacterial peptide, natural, bovine serum albumin(BSA), extracts of Chinese herbal medicine, hops extract, Liquid enzyme formulation fully dissolves, mixes, and is 5.2 by lactic acid regulation liquid compound enzyme pH value, stablizes 1.5h, simultaneously in electric-field intensity 40kV/cm, burst length 500 μ s, carry out high-pressure pulse electric (PEF) and be sterilized under the conditions of pulse frequency 350Hz, aseptic filtration, Sterile filling i.e. obtains liquid compound enzyme.
The liquid compound enzyme prepared through said method is 168CFU/mL in 12 months total plate counts of room temperature storage；At 40 DEG C Storing 12 months under the conditions of 0 DEG C, enzyme loss late alive is respectively 2% and 1%.
Embodiment 4
A kind of liquid compound enzyme for brewing, is prepared by the raw material of following parts by weight:
Liquid enzyme formulation 30 parts, hops extract 5 parts, sugar 5 parts, natural 4 parts, extracts of Chinese herbal medicine 5 parts, Antibacterial peptide 4 parts, thickener 3 parts, beta-schardinger dextrin 3 parts, polyalcohol 2 parts, glutathione 2 parts, metal chloride 1 part, ammonium sulfate 1 Part, cysteine 0.5 part, bovine serum albumin(BSA) 0.1 part.
Described liquid enzyme formulation is uniformly mixed by middle temperature amylase, dextranase, zytase 3:1:0.5 by volume；
Described sugar is that trehalose, GL-B, glucose 5:1:2 in mass ratio uniformly mixes；
Described natural is by grape pip procyanidin, Rosmarinus officinalis extract and apricot leaf extract 4:2 in mass ratio: 1 uniformly mixes；
Described thickener is uniformly mixed by carragheen, Arabic gum, gelatin, xanthans, CMC 3:2:1:1:0.5 in mass ratio Close；
Described polyalcohol is uniformly mixed by polyethylene glycol, glycerine, sorbierite, dithiothreitol (DTT) 8:4:2:1 in mass ratio；
Described metal chloride is zinc chloride, calcium chloride, sodium chloride, magnesium chloride, iron chloride 3:2:1:1:0.1 in mass ratio Uniformly mixing；
Described hops extract is prepared by embodiment 1；
Described extracts of Chinese herbal medicine is prepared by embodiment 2；
Preparation method is with embodiment 3.
The liquid compound enzyme prepared through said method is 208CFU/mL in 12 months total plate counts of room temperature storage；At 40 DEG C Storing 12 months under the conditions of 0 DEG C, enzyme loss late alive is respectively 2.2% and 1.3%.
Embodiment 5
A kind of liquid compound enzyme for brewing, is prepared by the raw material of following parts by weight:
Liquid enzyme formulation 50 parts, hops extract 15 parts, sugar 10 parts, natural 9 parts, extracts of Chinese herbal medicine 8 Part, antibacterial peptide 6 parts, thickener 5 parts, beta-schardinger dextrin 5 parts, polyalcohol 5 parts, glutathione 4 parts, metal chloride 3 parts, ammonium sulfate 2 parts, cysteine 1 part, bovine serum albumin(BSA) 0.3 part.
Described liquid enzyme formulation is uniformly mixed by middle temperature amylase, dextranase, zytase 4:1.5:1 by volume；
Described sugar is that trehalose, GL-B, glucose 8:3:4 in mass ratio uniformly mixes；
Described natural is by grape pip procyanidin, Rosmarinus officinalis extract and apricot leaf extract 6:4 in mass ratio: 3 uniformly mix；
Described thickener is uniformly mixed by carragheen, Arabic gum, gelatin, xanthans, CMC 5:4:3:2:1 in mass ratio；
Described polyalcohol is uniformly mixed by polyethylene glycol, glycerine, sorbierite, dithiothreitol (DTT) 10:6:4:3 in mass ratio Close；
Described metal chloride is zinc chloride, calcium chloride, sodium chloride, magnesium chloride, iron chloride 5:3:3:2:0.3 in mass ratio Uniformly mixing；
Described hops extract is prepared by embodiment 1；
Described extracts of Chinese herbal medicine is prepared by embodiment 2；
Preparation method is with embodiment 3.
The liquid compound enzyme prepared through said method is 214CFU/mL in 12 months total plate counts of room temperature storage；At 40 DEG C Storing 12 months under the conditions of 0 DEG C, enzyme loss late alive is respectively 2.5% and 1.7%.
Embodiment 6
A kind of liquid saccharifying enzyme for brewing, is prepared by the raw material of following parts by weight:
40 parts of carbohydrase, hops extract 10 parts, sugar 8 parts, natural 7 parts, extracts of Chinese herbal medicine 6 parts, anti- Bacterium peptide 5 parts, thickener 4 parts, beta-schardinger dextrin 4 parts, polyalcohol 3 parts, glutathione 3 parts, metal chloride 2 parts, 2 parts of ammonium sulfate, Cysteine 0.8 part, bovine serum albumin(BSA) 0.2 part.
Described sugar is that trehalose, GL-B, glucose 7:2:3 in mass ratio uniformly mixes；
Described natural is by grape pip procyanidin, Rosmarinus officinalis extract and apricot leaf extract 5:3 in mass ratio: 2 uniformly mix；
Described thickener is uniform by carragheen, Arabic gum, gelatin, xanthans, CMC 4:3:2:1.5:0.8 in mass ratio Mixing；
Described polyalcohol is uniformly mixed by polyethylene glycol, glycerine, sorbierite, dithiothreitol (DTT) 9:5:3:2 in mass ratio；
Described metal chloride is zinc chloride, calcium chloride, sodium chloride, magnesium chloride, iron chloride 4:2.5:2 in mass ratio: 1.5:0.2 uniformly mixes；
Described hops extract is prepared by embodiment 1；
Described extracts of Chinese herbal medicine is prepared by embodiment 2；
Preparation method is with embodiment 3.
The liquid saccharifying enzyme prepared through said method is 168CFU/mL in 12 months total plate counts of room temperature storage；At 40 DEG C Storing 12 months under the conditions of 0 DEG C, enzyme loss late alive is respectively 2% and 1%.
The biological stability of embodiment 7 liquid compound enzyme
With the liquid saccharifying enzyme of the embodiment of the present invention 6 preparation with the same specification liquid saccharifying enzyme of commercially available identical date of manufacture it is Subjects, under identical storage condition (room temperature, ventilation, lucifuge), uses " GB8726-2006 food additives carbohydrase system Agent " in detection method have detected the sanitary indexs such as pH value, total plate count, pathogenic bacteria, result such as table 1:
Table 1 stores 12 months microbiological indicator testing results
Result above shows: although commercial liquid carbohydrase shelf-life (3 months) interior Testing index is the most qualified, but when storing During to 6-12 month, its total plate count and coliform have exceeded standard, and content of microorganisms is incremented by rapidly, and pH value is down to from 5.5 3.6, decline substantially；Fluid present invention carbohydrase pollution microbes kind is few, and every Testing index stores and is superior to country in 12 months Standard, and along with the prolongation of the time of storage, slowly, pH value is almost without change, it is shown that liquid of the present invention for content of microorganisms propagation The biological stability that body compound enzyme is excellent.
Embodiment 8 liquid compound enzyme enzyme activity stability
The liquid saccharifying enzyme embodiment of the present invention 6 prepared divides with the same specification liquid saccharifying enzyme of commercially available identical date of manufacture Do not store 12 months under the conditions of 0 DEG C and 40 DEG C, use detection method in " GB8726-2006 food additives saccharidase preparation " Measuring the enzyme activity of liquid saccharifying enzyme, calculate enzyme loss late alive, enzyme loss late alive refers to actually detected enzyme activity and product mark The difference of note enzyme activity accounts for the percentage of mark enzyme activity, result such as table 2
Table 2 stores phase enzyme activity loss late
Result above shows, stores 12 months under the conditions of 0 DEG C and 40 DEG C, and fluid present invention carbohydrase is than commercial liquid sugar Changing the loss alive of enzyme enzyme and reduce by 19% and 98% respectively, have excellent temperature storage, enzyme activity is highly stable, long shelf-life.
Embodiment 9 present invention implements the liquid compound enzyme of 3 preparations application test in brewing
Australia barley moisture low (0.8-1.1) % in import barley, protein content the lowest the highest (9.45- 10.22%), germination percentage is high (98.7-99.3%), and unit weight, mass of 1000 kernel height, impurity content is low, than other import barley quality relatively Good, it is particularly suitable for brewageing quality beer.The liquid compound enzyme science of the embodiment of the present invention 3 preparation compounds dextranase, xylan Enzyme and middle temperature amylase, be particularly suitable for Australia Fructus Hordei Germinatus and add in the saccharification stage.
1. using method:
(1) raw material weight percentage composition: import Australia (Australian) Fructus Hordei Germinatus 40%, rice 20%, cracks rice 40%；
(2) the liquid saccharifying enzyme addition brew kettle that the embodiment of the present invention 3 is prepared by the saccharification stage:
(3) addition: every t dried malt adds 700ml；
Action condition: Fructus Hordei Germinatus wine with dregs optimum PH 5.2；Protein hydrolysis temperature 50 C, time 90min；Optimal material water quality ratio 1:5.Mixing converted mash (Fructus Hordei Germinatus wine with dregs mixes with gelatinization wine with dregs) optimum PH 5.4；Saccharification temperature 66 DEG C；Saccharificatinn period 60min.
2. using effect:
10 ° of light beer saccharification stage effectiveness tables of comparisons brewageed by table 3
Note: carbohydrase commercial liquid carbohydrase compound enzyme fluid present invention compound enzyme
Data analysis from table: when high adjunct ratio (60%) beer brewing, personnel, equipment, raw material, technique, water quality Deng under working condition same case, fluid present invention compound enzyme filtration time compared with common liq carbohydrase averagely shortens 38.28%, original wort concentration averagely improves 4.58%, and wheat juice turbidity averagely reduces by 68.83%, and α amino nitrogen content averagely improves 24.84%, wheat juice yield averagely improves 8.3%, and wheat juice yield and quality significantly improve.
10 ° of light beer fermentation stage effect comparison tables brewageed by table 4
Note: carbohydrase commercial liquid carbohydrase compound enzyme fluid present invention compound enzyme
Data analysis from table: when high adjunct ratio (60%) beer brewing, personnel, equipment, raw material, technique, water quality Deng under working condition same case, beer compound enzyme of the present invention volatile acid compared with ordinary beer compound enzyme averagely reduces 71.67%, aldehydes averagely reduces by 52.09%, and biacetyl averagely reduces by 51.78%, and the degree of fermentation averagely improves 7.08%, reclaims ferment Female death rate averagely reduces by 60%, and zymotic fluid quality significantly improves, and yeast growth and reproductive performance are obviously improved.
10 ° of light beer finished product effect comparison tables brewageed by table 5
Note: A commercially available ordinary beer carbohydrase B fluid present invention compound enzyme
Data analysis from table: the finished beer that high adjunct ratio (60%) brews, fluid present invention compound enzyme is with general Logical beer saccharification enzyme compares malt flavor and the coordination obvious, soft of hops fragrance, and foaming properties is excellent, and bubble the holding property time is long；Turbid Spending relatively low, diacetyl content is relatively low, and fermentation is good, and alcoholic strength is higher, finished beer either organoleptic indicator or physical and chemical index Have precedence over the most far away ordinary beer compound enzyme, meet or exceed national existing beer top grade standard.

Claims (5)

1. for a liquid compound enzyme for brewing, prepare the raw material of following parts by weight:
Liquid enzyme formulation 30-50 part, hops extract 5-15 part, sugar 5-10 part, natural 4-9 part, Chinese herbal medicine carries Take liquid 5-8 part, antibacterial peptide 4-6 part, thickener 3-5 part, beta-schardinger dextrin 3-5 part, polyalcohol 2-5 part, glutathione 2-4 part, gold Belong to chloride 1-3 part, ammonium sulfate 1-2 part, cysteine 0.5-1 part, bovine serum albumin(BSA) 0.1-0.3 part；
The preparation method of described hops extract is: is put by hops in supersonic wave cleaning machine and cleans 10-in 200W, 40KHz 15min, drains, and pulverizes immediately after-18-22 DEG C of freezing 1-2h, freezing thickness of feed layer 3-5cm, grinding particle size 0.5- 3mm, adds ethanol and the mixture of propyl alcohol pulverizing hops weight 1-3 times, and the mass ratio of ethanol and propyl alcohol mixing is 2-4: 1-3, is 3.0-3.5 with breast acid for adjusting pH value, carries out micro-under the conditions of power 150-300W, frequency 2000Hz, temperature 20-35 DEG C Ripple extracts 20-30min, simultaneously at power 200-300W, carries out ultrasonic assistant extraction under the conditions of frequency 30-40KHz；Insulation 1- 3h, then, carries out Microwave Extraction 15-20min, simultaneously at power 300-under the conditions of power 200-400W, frequency 2000Hz 500W, carries out ultrasonic assistant extraction under the conditions of frequency 40-50KHz, be finally naturally cooling to room temperature, filters, and filtrate is in electric field Intensity 35-45kV/cm, burst length 400-600 μ s, carry out high-pressure pulse electric sterilization under the conditions of pulse frequency 200-300Hz 20-30min i.e. obtains hops extract；
Described liquid enzyme formulation select free dextranase, zytase, middle temperature amylase, AMS, Thermostable α-Amylase, Fungal alpha-amylase, isoamylase, carbohydrase, beta amylase, Pullulanase, papain, bromelain, neutral protein Enzyme, acid protease, proline protein enzyme, ficin, 1,4 beta-glucanase, heatproof beta glucan complex enzyme, β-Portugal gather Sugar complex enzyme, mannonase pentosanase, acid phosphatase, lipase, phosphate, glucuroide, glucose isomerase Any one or several in the group of enzyme, tannase, lactase, catalase, alpha-acetolactate decarboxylase composition；
Described natural is grape pip procyanidin, Rosmarinus officinalis extract and apricot leaf extract 4-6:2-4 in mass ratio: 1-3 mixes；
The preparation method of described extracts of Chinese herbal medicine is: count by weight, accurately weighs Radix Astragali 60-70 part, Radix Angelicae Sinensis 55-65 Part, Radix Codonopsis 40-45 part, Radix Glycyrrhizae 40-45 part, cordate houttuynia 35-45 part, Divine Comedy 35-45 part, honeysuckle 25-35 part, Poria cocos 20-30 Part, polygala root 15-25 part, fry fennel 15-25 part, bighead atractylodes rhizome 10-20 part, bark of official magnolia 10-20 part；Respectively said herbal medicine is crushed to grain Footpath is less than 2 millimeters, then uniformly mixes and add the water of 3-6 times of weight in container, controls temperature 70 C-90 DEG C and keeps 2- 4h, is then cooled to 45-60 DEG C, and the mixing enzyme preparation adding mixed material gross weight 5-10% carries out enzymolysis, regulates with lactic acid PH value is 5.5-6.8, enzymolysis 2-4h, finally adds 0.5-3 times of w ethanol of mixed material and the mixture of propyl alcohol, ethanol and third The mass ratio of alcohol mixing is 1:1-2, controls temperature and keeps 3-4h to 60 DEG C-78 DEG C, filters, obtain the first filtrate；Add filter residue 1-3 The water of times weight, controls temperature 85 DEG C-95 DEG C and keeps 1-3h, be then cooled to 25-35 DEG C, filter, obtain the second filtrate；By first Filtrate and the second filtrate merge according to mass ratio 2-3:1-2, in electric-field intensity 35-45kV/cm, burst length 400-600 μ s, arteries and veins Carry out high-pressure pulse electric sterilization 20-30min under the conditions of rushing frequency 300-400Hz and i.e. obtain extracts of Chinese herbal medicine；
The parts by weight of described mixed enzyme consist of: endo-beta-glucanase 10-20 part, outer 1,4 beta-glucanase 10-20 part, β-grape Glycosidase 10-15 part, zytase 15-20 part, pentosanase 15-20 part, Pullulanase 20-30 part, beta amylase 10-15 Part, acid protease 10-15 part, pectase 10-15 part, seminase 10-15 part, glucose oxidase 5-10 part, acid phosphorus Acid enzyme 5-10 part, lipase 5-8 part；
Described concentration of alcohol >=75%；
Described polyalcohol is uniform by polyethylene glycol, glycerine, sorbierite, dithiothreitol (DTT) 8-10:4-6:2-4:1-3 in mass ratio Mixing；
Described metal chloride is zinc chloride, calcium chloride, sodium chloride, magnesium chloride, iron chloride 3-5:2-3:1-3:1-in mass ratio 2:0.1-0.3 uniformly mixes.
2. the liquid compound enzyme for brewing as claimed in claim 1, it is characterised in that described liquid enzyme formulation by Temperature amylase, dextranase, zytase 3-4:1-1.5:0.5-1 by volume uniformly mixes.
3. the liquid compound enzyme for brewing as claimed in claim 1, it is characterised in that described sugar is trehalose, spirit In sesame polysaccharide, glucose one or more.
4. the preparation method of the liquid compound enzyme for brewing as described in claim 1-3 is arbitrary, it is characterised in that bag Include following steps: in terms of parts by weight, the most accurately weigh thickener according to formula, add the sterilized water of its quality 4-6 times, room Temperature soaks 3-8h, and intensification limit, limit is stirred, 80-90 DEG C of insulation 20-30min, and stirring is fully dissolved, continued insulation, depend on while stirring Secondary addition beta-schardinger dextrin, polyalcohol, sugar, metal chloride, be then cooled to 25-35 DEG C, be sequentially added into while stirring ammonium sulfate, Cysteine, glutathione, antibacterial peptide, natural, bovine serum albumin(BSA), extracts of Chinese herbal medicine, hops extract, Liquid enzyme formulation fully dissolves, mixes, and is 2.5-6 by lactic acid regulation liquid compound enzyme pH value, stablizes 0.5-2h, simultaneously in electric field Intensity 35-45kV/cm, burst length 400-600 μ s, carry out high-pressure pulse electric sterilization under the conditions of pulse frequency 300-400Hz, Aseptic filtration, sterile filling i.e. obtain liquid compound enzyme.
5. the application in brewing of the liquid compound enzyme for brewing as described in claim 1-3 is arbitrary.
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