CN107296189A - The preparation method and composite enzyme beverage of a kind of composite enzyme beverage - Google Patents
The preparation method and composite enzyme beverage of a kind of composite enzyme beverage Download PDFInfo
- Publication number
- CN107296189A CN107296189A CN201710452098.8A CN201710452098A CN107296189A CN 107296189 A CN107296189 A CN 107296189A CN 201710452098 A CN201710452098 A CN 201710452098A CN 107296189 A CN107296189 A CN 107296189A
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- Prior art keywords
- fermentation
- composite enzyme
- enzyme beverage
- preparation
- stoste
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- 238000002360 preparation method Methods 0.000 title claims abstract description 21
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims abstract description 123
- 238000000855 fermentation Methods 0.000 claims abstract description 108
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- 241000894006 Bacteria Species 0.000 claims abstract description 51
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims abstract description 50
- 239000004310 lactic acid Substances 0.000 claims abstract description 25
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- 241000235342 Saccharomycetes Species 0.000 claims abstract description 13
- 229940088598 enzyme Drugs 0.000 claims description 70
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- 238000000034 method Methods 0.000 claims description 16
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- 241000228347 Monascus <ascomycete fungus> Species 0.000 description 1
- 108010038807 Oligopeptides Proteins 0.000 description 1
- 102000015636 Oligopeptides Human genes 0.000 description 1
- 241001248610 Ophiocordyceps sinensis Species 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
- A23L2/382—Other non-alcoholic beverages fermented
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/52—Adding ingredients
- A23L2/60—Sweeteners
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/123—Bulgaricus
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/21—Streptococcus, lactococcus
- A23V2400/249—Thermophilus
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Mycology (AREA)
- Non-Alcoholic Beverages (AREA)
Abstract
The invention belongs to technical field of food beverage, more particularly to the preparation method and composite enzyme beverage of a kind of composite enzyme beverage, the cereal such as brown rice and fruits and vegetables are used for main material, utilize saccharomycete, acetic acid bacteria and lactic acid bacteria carry out 3 step fermentations, in the different phase of manufacturing process, using different strain step fermentation, to meet demand of the human body to ferment in terms of health and nutritive effect, in addition to the vitamin and minerals of fermentation process except retaining fruits and vegetables etc., particularly coarse food grain by fermentation after, insoluble diedairy fiber therein can be made to resolve into easily by human consumption, the characteristics of with nutritious and unique flavor.
Description
【Technical field】
The invention belongs to the preparation method and compound ferment of technical field of food beverage, more particularly to a kind of composite enzyme beverage
Plain beverage.
【Background technology】
The process of enzyme beverage is prepared by raw material of fruits and vegetables can promote carbohydrate in fruit and vegetable materials, protein, fat
The metabolic response of the materials such as class, it is a large amount of produce and accumulation primary and secondary metabolite, generation organic acid, oligosaccharide, sugar alcohol,
Enzyme, oligopeptide.The multiple beneficial such as polyphenol composition can promote the propagation of intestinal beneficial bacterium, the breeding of suppression harmful bacteria and corruption
The formation of material, plays regulation gut flora balance, enhancing is immune, promote the effect such as sleep, anti-aging, has a variety of to human body
Beneficial functional.Thus enzyme beverage or Juice fermented beverage are prepared because of its alimentary health-care function and uniqueness using natural fruit and vegetables
Local flavor is favored by consumers in general deeply.
The country is fermented such as Publication No. by raw material of fruits and vegetables more than preparation enzyme beverage using single fruits and vegetables at present
" CN 102356899A " patent of invention discloses one kind and Fructus Chaenomelis ferment is prepared with single pawpaw or single culture or compound bacteria is used
Plant and carry out single step fermentation；" CN 102018259A " patent of invention discloses a kind of using highland barley monascus single step fermentation Publication No.
Cordyceps sinensis enzyme beverage；" CN 101341995A " patent of invention discloses a kind of using Lactobacillus plantarum and Bao Jiali Publication No.
The beverage of sub- lactobacillus mixing single step fermentation vegetable juice, it is short to there is fermentation time in above-mentioned patent, the simple shortcoming of fermentation process,
Fermented using single fruits and vegetables, it is impossible to meet human body to ferment health and nutritive effect in terms of demand, and it is therein not
Soluble dietary fiber is not easily decomposed by human consumption.
【The content of the invention】
For solve prior art enzyme beverage more than fermented using single fruits and vegetables, it is impossible to meet human body and ferment existed
The problem of demand in terms of health and nutritive effect, the present invention provides a kind of preparation method of composite enzyme beverage, using brown rice
It is main material Deng cereal and characteristic fruits and vegetables, carries out 3 step fermentations using saccharomycete, acetic acid bacteria and lactic acid bacteria, in manufacturing process not
In the same stage, using different strain step fermentation, to meet demand of the human body to ferment in terms of health and nutritive effect, fermented
Journey is outer except the vitamin and minerals etc. for retaining fruits and vegetables, particularly coarse food grain by fermentation after, insoluble meals fibre therein can be made
Dimension is resolved into easily by human consumption, the characteristics of with nutritious and unique flavor.
Another object of the present invention is to provide a kind of composite enzyme beverage.
The present invention is the technical scheme that is used of its technical problem of solution：
A kind of preparation method of composite enzyme beverage, comprises the following steps：
(1) Feedstock treating：Add water defibrination in proportion after cereal is impregnated, and stoste is made；
(2) toward alpha-amylase is added in stoste, stoste is liquefied；
(3) it is saccharified toward the stoste addition beta amylase after liquefaction；
(4) saccharomycete is added toward the stoste after saccharification, carries out saccharomycetes to make fermentation, alcohol fermentation liquid is made；
(5) acetic fermentation is carried out to alcohol fermentation liquid, stops acetic fermentation when acetic acid content no longer rises, be made
Acetic acid fermentation liquid；
(6) toward addition fruit and vegetable materials and lactic acid bacteria in acetic acid fermentation liquid, lactic fermentation is carried out, lactic fermentation liquid is made.
Preferably, the cereal includes brown rice, oat and corn, sets the pH value range 6.0-7.0 of stoste.
Preferably, when being saccharified in step (3), the enzyme concentration of beta amylase is 6u/g, and saccharification temperature is 40 DEG C, saccharificatinn period
For 40min, the pH for concurrently setting stoste is 4.5.
Preferably, in step (4) during saccharomycetes to make fermentation, yeast consumption is 9%, and fermentation time is 2h, and fermentation temperature is 35
DEG C, GSH contents are 2.73mg/g.
Preferably, in step (5) during acetic fermentation, choose acetic acid bacteria Shanghai and make 1.01 as starting strain, during acetic fermentation
Between be set to 84h.
Preferably, the fruit and vegetable materials include dragon fruit, pawpaw, mango, pineapple and strained tomatoes.
Preferably, the lactic acid bacteria is from Bulgarian lactic acid bacterium and streptococcus thermophilus.
Preferably, during the lactobacillus-fermented, lactic acid consumption is 8%, and fermentation temperature is 40 DEG C, and fermentation time is 35h,
GSH contents are 3.25mg/g.
Preferably, an allocation process is also included after step (6), sweetener is added toward the lactic fermentation liquid.
Compared with prior art, the present invention has the following advantages：
1st, the present invention uses the cereal such as brown rice and fruits and vegetables for main material, and 3 steps are carried out using saccharomycete, acetic acid bacteria and lactic acid bacteria
Fermentation, in the different phase of manufacturing process, using different strain step fermentation, to meet human body to ferment in health and nutrition work(
The demand in efficacious prescriptions face, fermentation process in addition to retaining vitamin and minerals etc. of fruits and vegetables, particularly coarse food grain by fermentation after, can make
Insoluble diedairy fiber therein is resolved into easily by human consumption, the characteristics of with nutritious and unique flavor；
2nd, present invention may determine that going out the optimum process condition of enzyme beverage production, the composite enzyme drink of sweet and sour taste is produced
Material, and the main matter of influence function can be illustrated, the present invention can promote fruit process industry to develop, and improve the work of enzyme beverage production
Skill technical merit, can also provide special bacteria and technique for production, increase product variety, widen realm of sale, meet consumption city
The demand of field；
3rd, the development of enzyme beverage of the present invention, meets the trend that domestic and international food develops to functionalization direction, has
Very big market potential, makes full use of the production of cereal and fruits and vegetables development of resources ferment product, has both met the industrial policy of country,
The marketing problem of local fresh fruit is partly solved again, with very high Social benefit and economic benefit；
4th, the present invention can consume substantial amounts of fruits and vegetables every year by the fermentation of Cereals and characteristic fruits and vegetables, and this is to releasing orchard worker
Sell fruit problem, reduce because fruit is unsalable and the economic loss brought of rotting, be that Liao Yitiaoxin roads are started in increasing peasant income.
【Brief description of the drawings】
Technical scheme in order to illustrate the embodiments of the present invention more clearly, makes required in being described below to embodiment
Accompanying drawing is briefly described, it should be apparent that, drawings in the following description are only some embodiments of the present invention, for
For those of ordinary skill in the art, on the premise of not paying creative work, other can also be obtained according to these accompanying drawings
Accompanying drawing.
Fig. 1 is the process chart of the present invention；
Fig. 2 is the change schematic diagram that content of starch of the present invention is determined；
Fig. 3 is the change schematic diagram of influence of the pH value of the present invention to alpha-amylase activity；
The change schematic diagram of influence of the enzyme concentration to DE values when Fig. 4 is raw material gelatinization of the present invention；
The change schematic diagram of influence of the temperature to DE values when Fig. 5 is rice gelatinization of the present invention；
The change schematic diagram of influence of the enzymolysis time to DE values when Fig. 6 is rice gelatinization of the present invention；
The change schematic diagram of influence of the amount of water to DE values when Fig. 7 is rice gelatinization of the present invention；
Fig. 8 is the comparison schematic diagram of the acetic acid bacteria acid producing ability of the present invention；
Fig. 9 is the change procedure schematic diagram of acidity and wine degree during acetic fermentation of the present invention.
【Embodiment】
The invention will be further described below in conjunction with the accompanying drawings and the specific embodiments.
Accompanying drawing 1 is refer to, a kind of preparation method of composite enzyme beverage comprises the following steps：
(1) Feedstock treating：By the defibrination that added water in proportion after equiponderant brown rice, oat, corn hybrid infusion, original is made
Liquid, concurrently sets in the pH value range 6.0-7.0 of stoste, the present embodiment pH being adjusted to 6.5；
(2) toward alpha-amylase is added in stoste, stoste is liquefied, the enzyme concentration for concurrently setting alpha-amylase is 10u/
G, condensing temperature is 80 DEG C, and liquefying time is 30min, and material-water ratio is 1: 4；This step specific implementation process is as follows：
Liquefy material requested and instrument
HH-4 thermostat water bath Weirs Instrument Ltd.；HC-TPH-5 electronic balances Shanghai analytical instrument factory；722S types
Spectrophotometer Shanghai analytical instrument factory；Alpha-amylase (2 × 104U/g) extensive and profound in meaning star biotechnology Co., Ltd, iodine, iodine
It is that analysis is pure to change potassium etc.；Brown rice, oat, corn etc. are purchased from Zhongshan city sun city supermarket of Wal-Mart.
Experimental method
1.1 starch standard curves are determined
1.000g is accurately weighed with assay balance and analyzes pure starch, 5.0mL distilled water is added and homogenate is made, be gradually poured into
In the distilled water of 90mL or so boilings, bevelling stirring in side produces the gelatinization of starch solution of clear, put in 100mL volumetric flasks,
With a small amount of distilled water flushing beaker, constant volume, this starch solution concentration is 10mg/mL (A liquid).Draw A liquid 2.0mL and put 100mL appearances
Measuring bottle, constant volume, now starch concentration is 200ug/mL (B liquid).Tool plug scale test tube 8 is taken, standard starch solution is separately added into
0th, 0.5,1.0,1.5,2.0,2.5,3.0,4.0ml, and I-KI solution 2ml, then add distilled water to supply every test tube solution
10mL, shakes up, and makes after the stable 10min of blue solution, its extinction value is surveyed at 722 spectrophotometer 660nm wavelength.With extinction value
For ordinate, it is known that the concentration of starch solution is that abscissa draws standard curve, refer to accompanying drawing 2, and the figure is to be measured through experiment
The change schematic diagram that content of starch is determined.Obtaining calibration curve equation according to the figure is：
Y=0.089x+0.0630 coefficient Rs2=0.9962.
The measure of starch in 1.2 raw material dextrins
Pipette samples liquid 2.0mL, puts in 100mL volumetric flasks, uses distilled water constant volume, mixes.Really draw 2mL samples in quasi- Huaihe River
Solution (uptake becomes according to starch concentration in sample), puts 15mL tool plug scale test tubes, adds I-KI solution 0.2mL, until molten
Transparent blue is presented in liquid, and 10mL is supplied with distilled water, is mixed, and stands 10min, and extinction value is determined at 660nm wavelength by marking
Directrix curve finds content of starch in sample (g/100g).
The measure of 1.3 rice dextrin total sugar contents
Standard GB/T 9695.31-1991.
1.4DE values are calculated
DE=total reducing sugars/starch
The determination of optimum process condition：Single factor experiments are carried out with enzyme concentration, temperature, time, material-water ratio this four factors,
Determine the level of orthogonal test.According to single factor test result, using L9(3)4Orthogonal is carried out, result is analyzed simultaneously
It is determined that optimal hydrolysis result.
1.5 enzymolysis experiment of single factor results
1.5.1a- amylase optimal pH is determined
The activity that then cushioning liquid that pH is 4,5,6,7,8,9,10 determine a- amylase is respectively configured.
The measure of a- amylase activities：Iodine is faded away in the specific reaction of bluish violet using starch, in reddish brown
Color, the speed that its color disappears is relevant with enzymatic activity, can calculate enzyme activity, enzyme by fixed reacted absorbance (660nm)
Vigor is defined:1g solid enzyme powders or 1ml enzyme liquids the 1h liquefaction 1g soluble starches under the conditions of 60 degree, pH 6.0 are 1 enzyme activity
Unit of force.Accompanying drawing 3 is refer to, the figure is the change schematic diagram that influence of the pH value to alpha-amylase activity is measured through experiment, according to
The figure can be seen that the optimal pH of a- diastatic activities between 6.0-7.0.
1.5.2 the influence research that enzyme concentration is gelatinized to raw material
80 DEG C, 30min, 1: 3 material-water ratio, using content of starch as index, research enzyme concentration 5u/g, 10u/g, 15u/g,
The situation of change of DE values determines optimal enzyme concentration scope when 20u/g, 25u/g, 30u/g.Accompanying drawing 4 is refer to, the figure is through experiment
The change schematic diagram of influence of the enzyme concentration to DE values is measured, be can be seen that by the figure, when enzyme addition increase, the shallow lake in dextrin
Powder content reduction is obvious, and the increase of pol is obvious.After enzyme concentration reaches 20u/g, gelatinization tend to relax, and dextrin produce compared with
Many foams, it is unfavorable to producing, it is optimal enzyme concentration scope from 5~15u/g of enzyme concentration in summary.
1.5.3 the influence research that temperature is gelatinized to rice
Enzyme concentration be 10u/g, 30min, 1: 3 material-water ratio, using content of starch as index, research 60 DEG C, 70 DEG C, 80 DEG C, 90
DEG C, content of starch under the conditions of 100 DEG C, sugar content, the situation of change of DE values, it is determined that optimal hydrolysis temperature.
The activity of enzyme is limited by temperature, and when temperature is in optimum condition, hydrolysis result is best.Accompanying drawing 5 is refer to, should
Figure is the change schematic diagram that influence of the temperature to DE values is measured through experiment, be can be seen that by the figure, with the rise of temperature, heat-resisting shallow lake
Powder enzymatic activity is also increased to content of starch and acutely declined, and total sugar content rises rapidly.Hydrolysis result is most when temperature reaches 80 DEG C
Good, then DE increases tend towards stability, so 80 DEG C of temperature ranges of selection carry out orthogonal experiment.
1.5.4 the influence research that the time is gelatinized to rice
According to 80 DEG C of reaction solution setting, enzyme-added 10U/g, 1: 3 material-water ratio, using content of starch as index, research 10min,
Content of starch under the conditions of 20min, 30min, 40min, 50min, sugar content, DE value changes situations, it is determined that optimal enzymolysis time.
Digestion time is longer, and enzyme effect is bigger thus also better to the discomposing effect of starch in the chance of substrate.It refer to
Accompanying drawing 6, the figure is to measure the change schematic diagram of influence of the enzymolysis time to DE values through experiment, from the map analysis, with when
Between increase, the increase of DE values is obvious, when being tended towards stability after 40min, so selection time is entered in 20min, 30min, 40min scope
Row orthogonal experiment.
1.5.5 the influence research that material-water ratio is gelatinized to rice
80 DEG C, 30min, 10u/g enzyme concentration, using content of starch as index, during research material-water ratio 1: 2,1: 3,1: 4,1: 5
The situation of change of DE values, it is determined that optimal material-water ratio scope.
Appropriate material-water ratio, starch enzyme-to-substrate touch opportunity increase, hydrolysis result is preferable, when material moisture is very high
When, the chance of this contact can be significantly smaller, thus hydrolysis result decrease.Accompanying drawing 7 is refer to, the figure is to measure to add water through experiment
The change schematic diagram of the influence to DE values is measured, from the map analysis, best results, therefore choosing are gelatinized at 1: 2,1: 3,1: 4
Select these three levels and carry out orthogonal experiment.
The determination of alpha-amylase fermentation condition
The factor level table of the alpha-amylase fermentation condition of table 1
According to table 1, the Orthogonal experiment results such as table 2 of design, alpha-amylase fermentation condition Orthogonal experiment results table
The alpha-amylase fermentation condition Orthogonal experiment results table of table 2
Optimal gelatinization condition is A as can be seen from Table 22B2C2D3Combination.As enzyme concentration is 10u/g, and temperature is 80
DEG C, action time, optimal material-water ratio was 1: 4 to be 30min.From table it can also be seen that influence order of each factor to result
For:C>B>A>D, that is, the time and temperature being gelatinized is the maximum factor of influence.Enzyme concentration and material-water ratio influence are smaller.
(3) it is saccharified toward the stoste addition beta amylase after liquefaction, the enzyme concentration for concurrently setting beta amylase is 6u/g,
Saccharification temperature is 40 DEG C, and saccharificatinn period is 40min, and the pH of stoste is 4.5；This step specific implementation process is as follows：
The beta amylase selected that is saccharified is provided by Tianjin enzyme preparation factory, and vigor is 90,000 u/g.Vigour-testing method is：For:
The soluble starch of effect 1% under standard conditions, the enzyme activity of generation lmg maltose per minute is a unit of activity.
The determination of beta amylase fermentation condition
The factor level table of the beta amylase fermentation condition of table 3
According to table 3, the Orthogonal experiment results such as table 4 of design, beta amylase fermentation condition Orthogonal experiment results table
The beta amylase fermentation condition Orthogonal experiment results table of table 4
Optimal saccharificationization condition combines for A2B2C2D2 as can be seen from Table 4.As enzyme concentration is 6u/g, and temperature is
40 DEG C, action time, optimal pH was 4.5 to be 40min.
Raw material improves the utilization rate of raw material, reduces cost, can improve factory of enterprise after gelatinization and saccharification processing
The profit of family.
(4) saccharomycete is added toward the stoste after saccharification, carries out saccharomycetes to make fermentation, it is 9% to concurrently set yeast consumption, hair
The ferment time is 2h, and fermentation temperature is 35 DEG C, and GSH contents are 2.73mg/g, and alcohol fermentation liquid is made；This step specific implementation process
It is as follows：
Material requested：Saccharomycete includes Angel high activity dried yeast.
Instrument and equipment：Electric drying oven with forced convection, incubator, visible spectrophotometer, centrifuge, glutathione kit,
Vacuum packing machine.
Method：
The measure of reduced glutathione (GSH)：Glutathione RNA isolation kit.
Reduced glutathione (GSH) is a kind of active bio peptide with important physiological function, with functional activity thing
Matter-glutathione is index, carries out the experiment of the horizontal quadrature of four factor three, finally determines yeast consumption, fermentation time and fermentation
Temperature.
The determination of yeast optimal conditions of fermentation
The reacted Cereals of liquefying-saccharifying is transferred to round, tested by the condition of setting, when alcoholic strength is about
10% stopping fermentation.As a result it is as follows：
Reduced glutathione (GSH) is a kind of active bio peptide with important physiological function, with functional activity thing
Matter --- glutathione is index, carries out the experiment of the horizontal quadrature of four factor three.
The factor level table of the yeast fermentation condition of table 5
According to table 5, the Orthogonal experiment results such as table 6 of design, the orthogonal design table of yeast fermentation condition
The orthogonal design table of the yeast fermentation condition of table 6
Reduced glutathione (GSH) is a kind of active bio peptide with important physiological function, with functional activity thing
Matter --- glutathione is index, may determine that the primary and secondary that each factor influences on product Glutathione peptide content is suitable by table 6
Sequence is:A>B>C, i.e. yeast consumption>Fermentation time>Fermentation temperature.Optimum combination is A3B1C3, i.e., when yeast consumption be 9%,
When fermentation time is that 2h, fermentation temperature are 35 DEG C, the glutathione content highest in brown rice enzyme reaches 2.73mg/g.
(5) acetic fermentation is carried out to alcohol fermentation liquid, chooses acetic acid bacteria Shanghai and make 1.01 as starting strain, during acetic fermentation
Between be set to 84h, stop acetic fermentation when acetic acid content no longer rises, acetic acid fermentation liquid be made；This step is embodied
Process is as follows：
Acetic fermentation
The screening and domestication culture of acetic acid bacteria
Acetic acid bacteria Shanghai is made 1.01, Shen wheat 112-7#, Pasteur's acetobacter AS1.41 be inoculated in the sour test medium I of production, 30
DEG C 120rpm is cultivated 6 days, surveys its total acid content；Experimental result refer to accompanying drawing 8, and the figure is to measure the sour energy of acetic acid bacteria production through experiment
The comparison schematic diagram of power, it can be seen that 1.01 production acid amount maximums are made in acetic acid bacteria Shanghai, reaches 3.51%；Next to that AS1.41,
Total yield acid amount is that 3.32%. Shen wheat 112-7# and Pasteur's acetobacter are all smaller.Therefore it may determine that acetic acid bacteria Shanghai makes 1.01 in height
Acid producing ability under alcohol concentration is most strong, so selection acetic acid bacteria Shanghai, which makes 1.01, is used as starting strain.
The domestication culture of acetic acid bacteria：It is starting strain to make 1.01 acetic acid bacterias from Shanghai, by 1 0g glucose, 2g yeast extracts,
4mL soya-bean polypeptides solution is dissolved separately in 20 0mL distilled water, is sub-packed in 2 triangular flasks, in 121 DEG C of sterilizings 3
0min, is cooled to 30 DEG C, aseptically every bottle addition 4mL 9 5% alcohol, and access Shanghai make 1.01 acetic acid bacteria strains,
In the 4h of 29 DEG C of one 30 DEG C of 8h of shaken cultivation 1~2, pure training acetic acid bacteria seed liquor is obtained；The domestication culture of acetic acid bacteria, in sterilizing
The special yeast domestication nutrient solution of access 5.0% in composite fruit juice afterwards, 7 2h are cultivated at 28 DEG C~30 DEG C, karusen
Alcoholic strength is up to 5% or so, and then the acetic acid bacteria seed liquor of inoculation 5%, cultivates 7d at 29 DEG C~3 1 DEG C, obtain acetic acid bacteria the 1st
Generation domestication seed liquor, then in the same way through the 2nd, 3,4,5 be commissioned to train support after, gained acetic acid bacteria can be preferably in cider
Growth and breeding in smart zymotic fluid, the strain can be used as fruit vinegar acetic fermentation special bacteria and use, and preservation is used.
Acetic acid bacteria test tube slant culture medium：Yeast extract 1%, glucose 1%, nutrient agar 3.3%, calcium carbonate 2%, (9
5%) ethanol 3%.Culture medium is configured according to actual needs, is put into sterilizing (1 21 DEG C, 3 0min) in autoclave, is put into after sterilizing
Inclined-plane is standby.Slant strains should cultivate 7 2h in 30 DEG C.
Acetic acid bacteria triangular flask spreads cultivation culture medium：Yeast extract 1%, glucose 1%, (9 5%) alcohol 3%.According to actual needs
Culture medium is configured, sterilizing (1 21 DEG C, 3 0mi n) in triangular flask is dispensed into, it is standby after cooling.
Acetic acid bacteria I and II seed culture medium：Composite fruit juice alcoholic fermented liquor after 90 DEG C of 3 0min of sterilizing.Inoculum concentration
For 10%.
Wine degree and acidity change of the reaction solution during acetic fermentation refer to accompanying drawing 9, and earlier fermentation, acid amount changes not
Greatly, increases slowly, this may be relevant with the inhibitory action of initial high alcohol content Dichlorodiphenyl Acetate bacterium, the fermentation in 48 to 84 hours
Mid-term is improved due to acetic acid bacteria vigor, and fermenting speed is fast, and production acid is fast, and ferment later stage, wine degree and acidity tend to a stationary value, this with
The highly acidity Dichlorodiphenyl Acetate bacterium in fermentation later stage suppresses relevant, with alcohol reduction acidity rise, acetic fermentation remitted its fury.In fermentation 84
Acidity reaches 4.3g/100mL during hour, continues to extend fermentation time, acetic acid content can decline on the contrary, therefore 84 hours are optimal
The acetic fermentation time.
(6) it is toward adding dragon fruit equal by weight, pawpaw, mango, pineapple, strained tomatoes and ratio in acetic acid fermentation liquid
1: 1 Bulgarian lactic acid bacterium and streptococcus thermophilus, carry out lactic fermentation, and it is 8% to concurrently set lactic acid consumption, and fermentation temperature is
40 DEG C, fermentation time is 35h, and GSH contents are 3.25mg/g, and lactic fermentation liquid is made；This step specific implementation process is as follows：
Instrument needed for lactic fermentation：Colloid mill, insulating box, high-pressure sterilizing pot, electric furnace, balance, capping machine, water-bath.
Using functional activity material-glutathione as index, three factors three are carried out after adding Juice in yeast fermentation broth
Horizontal quadrature is tested, and optimization lactobacillus-fermented condition is lactic acid bacteria consumption, fermentation time, fermentation temperature.Lactobacillus-fermented generation
Gastric acid secretion can be mitigated by thanking to product, suppress enteral spoilage organisms, have good stimulation to intestinal wall nerve, can promote human consumption
The secretion of enzyme and the wriggling of enteron aisle, so as to promote digesting and assimilating for food, and prevent just to secrete.Meanwhile, lactic acid bacteria thalline is in vivo
After being decomposed, after its active ingredient is absorbed by organisms, body immunity can be strengthened.In addition, lactic acid bacteria can also secrete raw in suppression
The antibiotic of pathogen, such as lactobacillus bulgaricus element have obvious bactericidal effect.
Key points for operation
Actication of culture
Because storage conditions inhibit the metabolism and breeding of thalline, make the vigor of strain very weak, therefore, must enter before
Row activation.Then activation is divided in clean test tube using the hot water dissolving of 9 times of injection in skimmed milk power and is placed in high pressure sterilization
Sterilized in pot, culture medium is made, then lactic acid bacteria is carried out to 3 or 4 Secondary Cultures in the medium.When pH value be 4.0~4.2,
Viable count is more than 106, and lactic acid acidity strain at 0.8%~1.0%, up to normal vital, is then placed in refrigerator and preserves standby
With.
The factor level table of the lactic fermentation condition of table 7
According to table 7, the Orthogonal experiment results such as table 8 of design, lactic acid fermented Orthogonal experiment results table
The lactic acid fermented Orthogonal experiment results table of table 8
Optimum combination is A as seen from Table 83B1C3, i.e., when lactic acid consumption be 8%, fermentation temperature be 40 DEG C when, fermentation
Time is the glutathione content highest in 35h, ferment, reaches 3.25mg/g.
From can be seen that the raw material after saccharification is handled with upper table 5-7 through saccharomycete, acetic acid bacteria and the hair of lactic acid bacteria
Ferment, makes the content of active bio peptide-reduced glutathione (GSH) of the physiological function in final fermented product reach most
Height, imparts the specific functionality of product.
(7) sterilizing, homogeneous, concentration, allotment：Lactic fermentation liquid adds the sweet tastes such as sugar alcohol, galactolipin after homogeneous is concentrated
Agent optimizes allotment, makes product sweet and sour taste；The liquid of fermentation out contains acetic acid and lactic acid, additionally containing micro apple
The organic acids such as tartaric acid, lactic acid, citric acid and butanedioic acid.Because acetic acid has extremely strong pungent taste, even across after ageing not yet
Preferably directly drink.Therefore this experiment adds a certain amount of sweetener, to relax the pungent taste of acetic acid, makes taste softer, assists
Adjust nature.This step specific implementation process is as follows：
The allotment of enzyme beverage
The enzyme beverage formula of table 9 and horizontally disposed table
According to table 9, the Orthogonal experiment results such as table 10 of design, enzyme beverage formula orthogonal design table
The enzyme beverage of table 10 is formulated orthogonal design table
The optimization formula that can obtain apple vinegar by the analysis result of table 10 is combined as A3B2C2D3, i.e. fermented liquid base-material adds
Dosage is 250ml/L, and galactolipin is 2%, and honey addition is 2%, fructose syrup addition 6 ‰
After experiment deployment the enzyme beverage product color of gained it is uniform, it is fermentation give off a strong fragrance, sour and sweet palatability, function
Composition glutathione content >=2.00mg/g.Glutathione have remove human body cell in free radical, can with human body
Noxious material such as combines and excreted at the function, is antioxidant important in human body, antidote and enhancing immune function of human body
Physiological activator, therefore the functional enzyme beverage produced using the technology of the present invention meets domestic and international food to work(
The trend that direction is developed can be changed, with very big market potential and very high Social benefit and economic benefit.
(8) filling, finished product.
A kind of composite enzyme beverage, the composite enzyme beverage is made up of the preparation method of above-mentioned composite enzyme beverage.Should
Enzyme beverage product color is uniform, free from admixture；Smell：There are fermentation fragrance, free from extraneous odour；Acidity：Sour and sweet palatability.Tissue morphology：Shape
State is uniform, free from admixture；Impurity：The exogenous impurity being visible by naked eyes；Glutathione is about 3.25mg/g.
The preparation method for the composite enzyme beverage that the present invention is provided, uses the cereal such as brown rice and fruits and vegetables for main material, utilizes
Saccharomycete, acetic acid bacteria and lactic acid bacteria carry out 3 step fermentations, in the different phase of manufacturing process, using different strain step fermentation, with
Meet demand of the human body to ferment in terms of health and nutritive effect, vitamin and minerals of the fermentation process except retaining fruits and vegetables etc.
Outside, particularly coarse food grain by fermentation after, insoluble diedairy fiber therein can be made to resolve into easily by human consumption, with nutrition
The characteristics of abundant and unique flavor.
The foregoing is only presently preferred embodiments of the present invention, not for limiting the scope implemented of the present invention, other it is all its
Principle and basic structure are identical or approximate with the present invention, within protection scope of the present invention.
Claims (10)
1. a kind of preparation method of composite enzyme beverage, it is characterised in that comprise the following steps：
(1) Feedstock treating：Add water defibrination in proportion after cereal is impregnated, and stoste is made；
(2) toward alpha-amylase is added in stoste, stoste is liquefied；
(3) it is saccharified toward the stoste addition beta amylase after liquefaction；
(4) saccharomycete is added toward the stoste after saccharification, carries out saccharomycetes to make fermentation, alcohol fermentation liquid is made；
(5) acetic fermentation is carried out to alcohol fermentation liquid, stops acetic fermentation when acetic acid content no longer rises, acetic acid is made
Zymotic fluid；
(6) toward addition fruit and vegetable materials and lactic acid bacteria in acetic acid fermentation liquid, lactic fermentation is carried out, lactic fermentation liquid is made.
2. a kind of preparation method of composite enzyme beverage according to claim 1, it is characterised in that：The cereal includes rough
Rice, oat and corn, set the pH value range 6.0-7.0 of stoste.
3. a kind of preparation method of composite enzyme beverage according to claim 1, it is characterised in that：It is saccharified in step (3)
When, the enzyme concentration of beta amylase is 6u/g, and saccharification temperature is 40 DEG C, and saccharificatinn period is 40min, and the pH for concurrently setting stoste is
4.5。
4. a kind of preparation method of composite enzyme beverage according to claim 3, it is characterised in that：Yeast in step (4)
When bacterium is fermented, yeast consumption is 9%, and fermentation time is 2h, and fermentation temperature is 35 DEG C, and GSH contents are 2.73mg/g, work as alcoholic strength
Stop fermentation when about 10%.
5. a kind of preparation method of composite enzyme beverage according to claim 1, it is characterised in that：Acetic acid in step (5)
During fermentation, choose acetic acid bacteria Shanghai and make 1.01 as starting strain, the acetic fermentation time is set to 84h.
6. a kind of preparation method of composite enzyme beverage according to claim 1, it is characterised in that：The fruit and vegetable materials bag
Include dragon fruit, pawpaw, mango, pineapple and strained tomatoes.
7. a kind of preparation method of composite enzyme beverage according to claim 1, it is characterised in that：The lactic acid bacteria is selected
Bulgarian lactic acid bacterium and streptococcus thermophilus.
8. a kind of preparation method of composite enzyme beverage according to claim 7, it is characterised in that：The lactobacillus-fermented
When, lactic acid consumption is 8%, and fermentation temperature is 40 DEG C, and fermentation time is 35h, and GSH contents are 3.25mg/g.
9. a kind of preparation method of composite enzyme beverage according to claim 8, it is characterised in that：After step (6) also
Including an allocation process, sweetener is added toward the lactic fermentation liquid.
10. a kind of composite enzyme beverage, it is characterised in that：The composite enzyme beverage is as described in any one of claim 1 to 9
A kind of preparation method of composite enzyme beverage is made.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109170438A (en) * | 2018-10-24 | 2019-01-11 | 北京农学院 | A kind of quinoa fermented beverage and preparation method thereof rich in γ-aminobutyric acid |
| CN109170438B (en) * | 2018-10-24 | 2021-08-31 | 北京农学院 | Quinoa fermented beverage rich in gamma-aminobutyric acid and preparation method thereof |
| CN109303222A (en) * | 2018-11-22 | 2019-02-05 | 江苏省农业科学院 | A kind of preparation method of sugar-free no-alcohol type health composite enzyme drink |
| CN115491280A (en) * | 2022-10-25 | 2022-12-20 | 贵州省中国科学院天然产物化学重点实验室(贵州医科大学天然产物化学重点实验室) | Roxburgh rose enzymatic fruit vinegar and preparation process and application thereof |
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Application publication date: 20171027 |