CN108865569A - A kind of semen sojae atricolor polypeptide wine and preparation method thereof - Google Patents
A kind of semen sojae atricolor polypeptide wine and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a kind of semen sojae atricolor polypeptide wines and preparation method thereof, prepare semen sojae atricolor polypeptide liquid using semen sojae atricolor proteolysis and then are fermented into, while burdock extract and ginkgo biloba extract are added in fermentation substrate.The semen sojae atricolor polypeptide wine color prepared using method of the invention is abundant, and mouthfeel is coordinated, and has good inoxidizability, and healthcare function is prominent, enriches China health liquor market, improves the added value that semen sojae atricolor utilizes.
Description
Technical field
The present invention relates to health liquor processing technique fields, and in particular to a kind of semen sojae atricolor polypeptide wine and preparation method thereof.
Background technique
Semen sojae atricolor is the soybean that epidermis is black, it is civil claim teata glycinis or equine beans, be a kind of dual-purpose of drug and food and nutrition is rich
Rich health care food materials, there is the laudatory title of the king in beans.As people are continuous to the nutritional ingredient of semen sojae atricolor and the research of functional materials
Deepen so that it in many industries all by different degrees of attention, especially in industries such as food, medicine, chemical industry, semen sojae atricolor and
Its derivative has all been widely used.Wherein in the food industry, the several functions of semen sojae atricolor and its extract are utilized
Matter, such as anti-aging, alleviation menopause symptom, pre- preventing bone rarefaction, breast cancer, senile dementia, cardiovascular disease, have developed
Many relevant health foods.Such as punishment Jian Hua, Hou Qiaozhi etc. add emulsifier and stabilization using semen sojae atricolor and jujube as primary raw material
Semen sojae atricolor jujube cream is made in agent, ferments under certain condition after inoculating lactic acid bacterium, semen sojae atricolor red date fermentation beverage is made.Zhang Linjing
It is equipped with fresh milk Deng with homemade semen sojae atricolor peptide, develops a kind of auxotype milk beverage.Zeng Rong, Dong Hua are strong etc. with semen sojae atricolor and Iron Guanyin
For raw material, by techniques such as immersion, defibrator process, filtering, compoundings, manufactured soymilk is in terms of mouthfeel, smell, structural state and color
It is good.Zhai Weiwei produces black bean soy using solid and liquid state fermentation technique, improves the nutritive value of soy sauce.Heilongjiang Academy of Agricultural Sciences
Soybean thus black soybean be major ingredient, a series of black soybean food has been researched and developed out, such as Guiyuan oral liquid and nutritious black bean
Paste etc..
With the raising of health liquor market penetration rate, health liquor is received by more and more consumers, before market development
Scape is optimistic.It is also higher and higher to the quality requirements of health care wine product with the enhancing that consumer healthcare realizes.Currently on the market
The method that health liquor mostly uses dipping, the kind using direct fermentation production health liquor is few, in particular by protease
The report that solution liquid direct fermentation prepares polypeptide wine is few.Development for beans polypeptide products, there is whey poly-peptides liquor and beans at present
Koumiss etc., but prepare semen sojae atricolor polypeptide liquid using semen sojae atricolor proteolysis and then be fermented into semen sojae atricolor polypeptide wine and do not have relevant report also.
Summary of the invention
For the above-mentioned prior art, the object of the present invention is to provide a kind of semen sojae atricolor polypeptide wines and preparation method thereof.
To achieve the above object, the present invention adopts the following technical scheme that：
The first aspect of the present invention provides a kind of preparation method of semen sojae atricolor polypeptide wine, includes the following steps：
(1) semen sojae atricolor albumen is digested, obtains semen sojae atricolor polypeptide liquid；
(2) fructus arctii is thinly sliced, is placed in the aqueous solution containing sodium citrate and impregnates 10-15min, it is dry, obtain pre- place
Fructus arctii sample after reason；The ethanol solution that 25-30 times of weight is added into pretreated fructus arctii sample carries out homogenate extraction, institute
The volumetric concentration for stating ethanol solution is 70-80%, and extraction voltage is 100V, and extraction time 45-55s, extraction time is 2 times,
Obtain fructus arctii extracting solution；By fructus arctii extracting solution microwave drying, burdock extract is obtained；
(3) ginkgo leaf crushed after being dried is added the water of 6-8 times of weight, extracted at 80-100 DEG C at 80-100 mesh fine powder
1-3h, filtering discard filter residue, and filtrate is concentrated and dried, and obtain ginkgo biloba extract；
(4) by semen sojae atricolor polypeptide liquid, burdock extract and ginkgo biloba extract mix, add sucrose, anhydrous sodium sulfite and
Water, sterilizing, as fermentation substrate；Adjusting the semen sojae atricolor content of peptides in fermentation substrate is 1.0%, and burdock extract content is 2.0-
4.0%, ginkgo biloba extract content is 1.0-3.0%, and cane sugar content 12-16%, anhydrous sodium sulfite content is 0.4-
0.6%；
(5) saccharomycete is accessed into fermentation substrate, is fermented 9 days at 26 DEG C；
(6) fermentation liquid is clarified using the method for centrifugation to get semen sojae atricolor polypeptide wine is arrived.
Preferably, in step (1), the specific method of the enzymatic hydrolysis is：Alcalase alkali protease is first used, is in temperature
60 DEG C, pH be 8.5 under conditions of hydrolyze semen sojae atricolor albumen；Flavourzyme flavor protease is added after 30min, is in temperature
50 DEG C, pH be 7.0 under conditions of digest 30min；Neutrase neutral proteinase is added later, the condition for being 50 DEG C in temperature
Lower enzymatic hydrolysis 1h.
Preferably, in step (2), fructus arctii is cut into the thin slice with a thickness of 0.3-0.5cm.
Preferably, in step (2), the mass concentration of sodium citrate is 2-4% in the aqueous solution containing sodium citrate.
Preferably, in step (2), the microwave drying control vacuum pressure is -0.1-0.05MPa, drying temperature 40-
50℃。
Preferably, in step (4), the semen sojae atricolor content of peptides adjusted in fermentation substrate is 1.0%, and burdock extract content is
4.0%, ginkgo biloba extract content is 2.0%, cane sugar content 16%, and anhydrous sodium sulfite content is 0.5%.
Preferably, in step (5), the access amount of the saccharomycete is the 1.2 ‰ of fermentation substrate weight.
Preferably, in step (5), the saccharomycete is the Angel Yeast of Angel Yeast Co., Ltd's production.
Preferably, in step (6), centrifugal rotational speed 3500r/min, centrifugation time 10min.
The second aspect of the present invention provides the semen sojae atricolor polypeptide wine that the above method is prepared.Semen sojae atricolor prepared by the present invention is more
Peptide wine clear, in light yellow, no precipitating or suspended matter, sense organ appraise are scored at 88.6 or more.
Beneficial effects of the present invention：
The semen sojae atricolor polypeptide wine color prepared using method of the invention is abundant, and mouthfeel is coordinated, and has good antioxygen
The property changed, healthcare function is prominent, enriches China health liquor market, improves the added value that semen sojae atricolor utilizes.
Detailed description of the invention
Fig. 1：Influence of the content of peptides to alcoholic strength；
Fig. 2：Influence of the content of peptides to soluble solid；
Fig. 3：Influence of the sugaring amount to alcoholic strength；
Fig. 4：Influence of the sugaring amount to soluble solid；
Fig. 5：Determining content of peptides standard curve.
Specific embodiment
It is noted that following detailed description is all illustrative, it is intended to provide further instruction to the application.Unless another
It indicates, all technical and scientific terms used herein has usual with the application person of an ordinary skill in the technical field
The identical meanings of understanding.
As the method that background technology part is introduced, and health liquor currently on the market mostly uses dipping, using direct
The kind that fermentation method produces health liquor is few, prepares the report of polypeptide wine not in particular by protein enzyme solution direct fermentation
It is more.Based on this, the object of the present invention is to provide a kind of semen sojae atricolor polypeptide wines, are fermented by semen sojae atricolor polypeptide liquid prepared by semen sojae atricolor proteolysis
It forms.
In one embodiment of the present invention, the preparation method of the semen sojae atricolor polypeptide wine provided includes the following steps：
(1) semen sojae atricolor albumen is digested, the specific method of the enzymatic hydrolysis is：Alcalase alkali protease is first used,
Semen sojae atricolor albumen is hydrolyzed under conditions of temperature is 60 DEG C, pH is 8.5；Flavourzyme flavor protease is added after 30min,
Temperature is 50 DEG C, pH digests 30min under conditions of being 7.0；Neutrase neutral proteinase is added later, is 50 DEG C in temperature
Under conditions of digest 1h, obtain semen sojae atricolor polypeptide liquid；
(2) fructus arctii is cut into the thin slice with a thickness of 0.3-0.5cm, is placed in the water for the sodium citrate that mass concentration is 2-4%
10-15min is impregnated in solution, it is dry, obtain pretreated fructus arctii sample；25- is added into pretreated fructus arctii sample
The ethanol solution of 30 times of weight carries out homogenate extraction, and the volumetric concentration of the ethanol solution is 70-80%, extracts voltage and is
100V, extraction time 45-55s, extraction time are 2 times, obtain fructus arctii extracting solution；By fructus arctii extracting solution microwave drying, obtain
Burdock extract；
(3) ginkgo leaf crushed after being dried is added the water of 6-8 times of weight, extracted at 80-100 DEG C at 80-100 mesh fine powder
1-3h, filtering discard filter residue, and filtrate is concentrated and dried, and obtain ginkgo biloba extract；
(4) by semen sojae atricolor polypeptide liquid, burdock extract and ginkgo biloba extract mix, add sucrose, anhydrous sodium sulfite and
Water, sterilizing, as fermentation substrate；Adjusting the semen sojae atricolor content of peptides in fermentation substrate is 1.0%, and burdock extract content is 2.0-
4.0%, ginkgo biloba extract content is 1.0-3.0%, and cane sugar content 12-16%, anhydrous sodium sulfite content is 0.4-
0.6%；
(5) saccharomycete is accessed into fermentation substrate, is fermented 9 days at 26 DEG C；
(6) fermentation liquid is clarified using the method for centrifugation, centrifugal rotational speed 3500r/min, centrifugation time is
10min to get arrive semen sojae atricolor polypeptide wine.
The present invention prepares semen sojae atricolor polypeptide using the method for multienzyme fractional hydrolysis, can make full use of different enzymes optimal pH and
Hydrolysis temperature improves the enzymatic hydrolyzation of semen sojae atricolor albumen, and the semen sojae atricolor polypeptide of preparation is made to have anti-oxidation characteristics.
In preparation method of the invention, the composition of fermentation substrate is the trophic component for influencing the semen sojae atricolor polypeptide wine of preparation, wind
The key point of taste, mouthfeel and function.Enzymolysis liquid containing semen sojae atricolor albumen in fermentation substrate of the invention, the albumen in enzymolysis liquid
Matter enzyme breaks down into amino acids and polypeptide provide essential nutriment for the growth and breeding of saccharomycete, promote yeast
Fermentation.Particularly, the present invention is also added to burdock extract and ginkgo biloba extract in fermentation substrate, and it is more can to reduce semen sojae atricolor
The bitter taste of peptide improves mouthfeel, keeps the flavour of fermented wine more pure and mild, while can also improve the oxidation resistance of fermented wine.This
Also contain a certain amount of anhydrous sodium sulfate in the fermentation substrate of invention, the hair of non-yeast substance can be inhibited to a certain extent
Ferment process is dominated entire fermentation and is carried out.Therefore, in fermentation substrate of the invention, each raw material component is an organic whole, is lacked
One can not.Inventor has found in R&D process, reduces any one of above-mentioned fermentation substrate raw material components, or with phase
It is replaced like the raw material of property, then the function and effect of fermentation substrate entirety significantly reduce；In the base of fermentation substrate of the invention
Other raw material components are further added by plinth, the overall effect of fermentation substrate does not improve significantly, or even has fermentation substrate
The case where overall effect reduces occurs.
The additional amount of each raw material in fermentation substrate is also very crucial, needed for the fermentation of saccharomycete and the growth metabolism of yeast
Nutriment is semen sojae atricolor polypeptide, and the increase of content of peptides promotes the quickening of the fermentation of saccharomycete, and the process of fermentation generates alcohol simultaneously
Cause the reduction (Fig. 1 and Fig. 2) of soluble solid.But when content of peptides is greater than 1%, the increase tendency of alcoholic strength and solvable
The reduction rate trend of property solid content all tends towards stability, and does not change significantly, and with the increase of content of peptides, alcoholic strength
There is no significant change with soluble solid content, at this moment if increase content of peptides instead to polypeptide wine clarification cause it is difficult and
It wastes raw material, therefore selecting content of peptides in fermentation substrate is 1%.
Sugaring amount is as shown in Figures 3 and 4 to the influence result of semen sojae atricolor polypeptide wine fermentation.As seen from the figure, the low situation of sugaring amount
Under, alcoholic strength also tends towards stability, and the content of soluble solid no longer declines when fermenting the 7th day, and fermentation process is close to tail
Sound, final polypeptide wine alcoholic strength are relatively low.When sugaring amount is 12%-16%, fermented wine degree and soluble solid contain
The variation tendency of amount reaches unanimity.When sugaring amount increases to 24%, starting fermentation rate is slower, is the shadow because by osmotic pressure
It rings, but after adaptation in a few days, the high advantage of sugar content starts to highlight in fermentation liquid, the speed of ferment middle is also because of sugar
Divide sufficient and obviously accelerate, the alcoholic strength variation in latter stage of fermenting is still very greatly.So comprehensively considering, according to the alcohol in fermentation latter stage
The variation tendency of degree and soluble solid content, therefore select 12%~16% sugaring amount more appropriate.
The process that the additional amount of anhydrous sodium sulfate mainly influences fermentation can be led if the additional amount of anhydrous sodium sulfate is very few
Cause the generation of non-yeast fermentation of materials process；If the additional amount of anhydrous sodium sulfate is excessive, the fermentation process of saccharomycete can be produced
It is raw to inhibit.Comprehensively consider, selects the additional amount of anhydrous sodium sulfate for 0.4-0.6% in fermentation substrate of the invention.
Some precipitatings or suspended matter can be contained in the fermentation liquid prepared after fermented, it is therefore desirable to carry out to fermentation liquid clear
Clearly to prepare semen sojae atricolor polypeptide wine.The present invention is clarified using the method for centrifugation, still, the anti-oxidation characteristics meeting of semen sojae atricolor polypeptide wine
There is different degrees of reduction with the increase and the extension of time of centrifugation rate.For guarantee semen sojae atricolor polypeptide wine clarity and
Antioxygenic property, the centrifugal condition that the present invention selects for：3500r/min is centrifuged 10min.
Since the property of fructus arctii polyphenol is unstable, easily it is degraded, and homogenate extraction is the high speed rotation by rotary head, it will
Drug is ground, this process can generate heat, therefore, is also possible to will cause the degradation of fructus arctii polyphenol even with formulation is dodged,
Influence the recovery rate of fructus arctii polyphenol.The present invention is found surprisingly that, before homogenate extraction, by the certain density citric acid of fructus arctii
Sodium water solution is soaked for a period of time, and can effectively improve the stability of polyphenols in fructus arctii, reduces it during the extraction process
It degrades, improves the recovery rate of fructus arctii polyphenol.The concentration and soaking time of sodium citrate aqueous solution are steady for fructus arctii polyphenol
Be for qualitative protecting effect it is very crucial, only suitable concentration and soaking time just can be to Polyphenols chemical combination in fructus arctii
The stability of object plays a protective role.The present invention has found that the quality of sodium citrate is dense in sodium citrate aqueous solution through test of many times
Degree is 2-4%, optimal to the protecting effect of the stability of polyphenol compound in fructus arctii when soaking time is 10-15min.
During sudden strain of a muscle mentions, the raising for extracting voltage can promote the rupture of material cell wall, be conducive to polyphenols and release
It is put into solution；But with the raising for extracting voltage, the temperature that will lead to extracting solution is improved, to accelerate the oxygen of polyphenol
Change.Through comprehensively considering, the present invention is set as 100V for voltage is extracted, and under the extraction voltage, on the one hand can guarantee Polyphenols object
Mass-energy is adequately discharged into solution, on the other hand can avoid the oxidation of polyphenol as far as possible.
Unlike other extracting method, it is to need to immerse cutter head in solvent that sudden strain of a muscle, which mentions, needs certain solvent levels,
In addition, material is crushed in high-speed rotation process, solution viscosity increases, and in order to improve ingredient diffusion velocity in the solution, needs
Increase solvent usage；When liquid volume added reaches a certain level, recovery rate growth trend thaws, but because excessive extraction is molten
Agent will increase the workload of subsequent processing.Comprehensively consider, the additional amount of selective extraction solvent of the present invention is pretreated ox
The 25-35 times of weight of burdock sample, wherein when the additional amount of Extraction solvent is 30 times of weight of pretreated fructus arctii sample,
Extraction effect is best.
In homogenate extraction, with the increase of extraction time, increase tendency is presented in the yield of fructus arctii polyphenol.It is between at the extraction
When 20s to 40s, polyphenol recovery rate grows steadily, and there is recovery rate within the scope of 40s to 50s is obviously substantially increased, more greater than after 50s
Phenol yield shows downward trend.The appearance of yield decline may be since the component through a long time high-speed friction of extractor generates
Fuel factor and molecules in solution high-speed motion bring heat make a part of unstable polyphenol decompose.In summary
Consider, select the extraction time of 50s study it is most appropriate.
In order to enable those skilled in the art can clearly understand the technical solution of the application, below with reference to tool
The technical solution of the application is described in detail in the embodiment of body.
Test material used in the embodiment of the present invention and comparative example is the test material of this field routine, can be passed through
Commercial channel is commercially available.
Alkali protease Alcalase, neutral proteinase Neutrase used in the present invention and flavor protease
Flavourzyme is purchased from letter Bioisystech Co., Ltd of Novi.
Embodiment 1：The preparation of semen sojae atricolor polypeptide wine
1. the preparation of semen sojae atricolor albumen：
(1) pretreatment of semen sojae atricolor
Semen sojae atricolor is chosen from market, is cleaned up, is removed the peel, peeling semen sojae atricolor is put into pulverizer and is crushed, is smashed it through
50 meshes install sieving black bean powder, are put into drying box stand-by.
(2) black bean powder degreasing
Semen sojae atricolor fat and some substances for being dissolved in organic reagent, semen sojae atricolor are sloughed from black bean powder using petroleum ether degreasing method
Powder and petroleum ether are with 1:10 ratios, are sealed with preservative film, stir 3h under room temperature on digital-display magnetic stirrer, and incline supernatant
(yellow, containing petroleum ether, water and fat etc.) adds young oil ether and continues to stir 3h, be then allowed to stand 5h or more, incline supernatant
Sediment fraction is taken out and is air-dried by liquid, until air-drying becomes white powder, re-dry is stand-by.
(3) semen sojae atricolor albumen is prepared
Weigh the degreasing black bean powder of certain mass, solid-to-liquid ratio 1:10 ratio is added distilled water, in digital display under room temperature
It is stirred on magnetic stirring apparatus, until degreasing black bean powder is uniformly mixed with distilled water.The pH meter corrected is reused, with 1mol/L's
The above-mentioned pH value of solution of NaOH solution tune is 8.0, is removed in bean powder not using 4000r/min speed centrifugation 15min again after stirring 30min
Soluble substance.Take the HCl solution tune supernatant of its supernatant 2mol/L pH value be 4.5, then under the conditions of 4000r/min from
Heart 15min makes protein be in isoelectric point state and agglomerates generation precipitating and sink.It finally takes out precipitating washing centrifugation twice, abandons
Supernatant is removed, gained precipitating is semen sojae atricolor albumen, then stand-by after freeze-drying.
2. the preparation of semen sojae atricolor polypeptide：
Semen sojae atricolor polypeptide is prepared using the method for multienzyme fractional hydrolysis, the optimal pH in view of Alcalase alkali protease is 7-
9, temperature is 55-70 DEG C；The optimal pH of Neutrase neutral proteinase is 5.5-7.5, and temperature is 55-70 DEG C；Flavourzyme
The optimal pH of flavor protease is 5.0-7.5, and temperature is 50-65 DEG C.
The 2% Alcalase alkali protease for first using substrate protein concentration, temperature is 60 DEG C, pH is that 8.5 hydrolysis are black
Legumin, pH value constantly decline, and the pH of solution drops to 7.0 or so after about 30min, at this time add substrate protein concentration 3%
Flavourzyme flavor protease, temperature be 50 DEG C in the case where digest 30min, add 2% Neutrase later
Neutral proteinase digests 1h under conditions of temperature is 50 DEG C, further increases its enzymatic hydrolyzation, and being made has anti-oxidation characteristics
Semen sojae atricolor polypeptide.
3. the preparation of burdock extract：
Fructus arctii is cut into the thin slice with a thickness of 0.3-0.5cm, is placed in the aqueous solution for the sodium citrate that mass concentration is 3%
15min is impregnated, it is dry, obtain pretreated fructus arctii sample；The second of 30 times of weight is added into pretreated fructus arctii sample
Alcoholic solution carries out homogenate extraction, and the volumetric concentration of the ethanol solution is 75%, and extractions voltage is 100V, extraction time 50s,
Extraction time is 2 times, obtains fructus arctii extracting solution；By fructus arctii extracting solution microwave drying, burdock extract is obtained.
4. the preparation of ginkgo biloba extract：
By ginkgo leaf crushed after being dried at 80-100 mesh fine powder, the water of 8 times of weight is added, extracts 2h at 100 DEG C, filters,
Filter residue is discarded, filtrate is concentrated and dried, and obtains ginkgo biloba extract.
5. the preparation of fermentation substrate：
Semen sojae atricolor polypeptide, burdock extract and ginkgo biloba extract are mixed, sucrose, anhydrous sodium sulfite and water is added, goes out
Bacterium, as fermentation substrate；Adjusting the semen sojae atricolor content of peptides in fermentation substrate is 1.0%, and burdock extract content is 4.0%, silver
Apricot extract content is 2.0%, cane sugar content 16%, and anhydrous sodium sulfite content is 0.5%；It is above weight percent
Than.
6. fermentation：
Angel Yeasts are accessed by 1.2 ‰ into fermentation substrate, are fermented 9 days at 26 DEG C；
7. clarification：
Fermentation liquid is clarified using the method for centrifugation, centrifugal rotational speed 3500r/min, centrifugation time 10min, i.e.,
Obtain semen sojae atricolor polypeptide wine.
Comparative example 1：
The composition of fermentation substrate is adjusted, the semen sojae atricolor content of peptides in fermentation substrate is 1.0%, cane sugar content 16%, nothing
Water sodium sulfite content is 0.5%.Angel Yeasts are accessed by 1.2 ‰ into fermentation substrate, are fermented 9 days at 26 DEG C；Using centrifugation
Method fermentation liquid is clarified, centrifugal rotational speed 3500r/min, centrifugation time be 10min to get arrive semen sojae atricolor polypeptide wine.
Comparative example 2：
The composition of fermentation substrate is adjusted, the semen sojae atricolor content of peptides in fermentation substrate is 1.0%, cane sugar content 16%.To
Angel Yeasts are accessed by 1.2 ‰ in fermentation substrate, are fermented 9 days at 26 DEG C；Fermentation liquid is clarified using the method for centrifugation, from
Heart revolving speed be 3500r/min, centrifugation time be 10min to get arrive semen sojae atricolor polypeptide wine.
Comparative example 3：
The composition of fermentation substrate is adjusted, the semen sojae atricolor content of peptides in fermentation substrate is 1.0%, cane sugar content 16%, nothing
Water sodium sulfite content is 0.5%.Angel Yeasts are accessed by 1.2 ‰ into fermentation substrate, are fermented 9 days at 26 DEG C；Fermentation liquid mistake
It filters to get semen sojae atricolor polypeptide wine is arrived.
Comparative example 4：
The composition of fermentation substrate is adjusted, the semen sojae atricolor content of peptides in fermentation substrate is 1.0%, and burdock extract content is
4.0%, cane sugar content 16%, anhydrous sodium sulfite content is 0.5%.Angel Yeasts are accessed by 1.2 ‰ into fermentation substrate,
It ferments 9 days at 26 DEG C；Fermentation liquid is clarified using the method for centrifugation, centrifugal rotational speed 3500r/min, centrifugation time is
10min to get arrive semen sojae atricolor polypeptide wine.
Comparative example 5：
The composition of fermentation substrate is adjusted, the semen sojae atricolor content of peptides in fermentation substrate is 1.0%, and ginkgo biloba extract content is
2.0%, cane sugar content 16%, anhydrous sodium sulfite content is 0.5%.Angel Yeasts are accessed by 1.2 ‰ into fermentation substrate,
It ferments 9 days at 26 DEG C；Fermentation liquid is clarified using the method for centrifugation, centrifugal rotational speed 3500r/min, centrifugation time is
10min to get arrive semen sojae atricolor polypeptide wine.
Test example 1：Alcohol content measurement, determining content of peptides and subjective appreciation in semen sojae atricolor polypeptide wine
1. alcohol content measures：
It takes 100mL wine sample in 500mL cucurbit with volumetric flask, adds 100mL water sum number grain bead, install condenser,
It is removed when slipping out liquid, about 95mL with the reception of 100mL volumetric flask, adds water to be settled to scale, shake up, liquid will be slipped out and pour into 100mL amount
In cylinder, it is gently put into thermometer and alcohol meter, until alcohol meter can float, and reads when static, then looks into《Alcohol is relatively close
Degree and the alcoholic strength table of comparisons》It is converted into alcoholic strength.
The alcohol content measurement result of semen sojae atricolor polypeptide wine prepared by embodiment 1, comparative example 1-3 is shown in Table 1.
Table 1：Alcohol content measurement result
| Product | Alcohol content (%) |
| Embodiment 1 | 9.8 |
| Comparative example 1 | 8.4 |
| Comparative example 2 | 7.5 |
| Comparative example 3 | 9.3 |
2. determining content of peptides：
(1) drafting of standard curve
Six test tubes are taken, are numbered respectively, reagent needed for being added according to table 2-4, it is necessary to reagent A first be added and create alkaline item
Then part adds biuret reagent B, it is 4mL that the dosage of biuret reagent, which has altogether, places 30min under room ambient conditions after mixing,
Then the ratio of its light absorption value of measurement at 540nm, A reagent and B reagent is 3:2, convenient for test substance and biuret B reagent
Reaction.Using bovine serum albumin content as abscissa, measuring light absorption value is ordinate, draws the regression equation (Fig. 5) of standard curve.
(2) measurement of semen sojae atricolor content of peptides
By 0.9mL distilled water, 2.4mL biuret reagent A is added and is added in the sample to be tested of 0.1mL, is added after mixing
1.6mL biuret reagent B, while doing three repetitions measures its absorbance value using blank reagent as reference respectively, calculate to
Test sample kind content of peptides.Content of peptides is obtained according to standard curve regression equation calculation.
The alcohol content measurement result of semen sojae atricolor polypeptide wine prepared by embodiment 1, comparative example 1-3 is shown in Table 2.
Table 2：Determining content of peptides result
| Product | Content of peptides (mg/mL) |
| Embodiment 1 | 9.28 |
| Comparative example 1 | 8.25 |
| Comparative example 2 | 7.64 |
| Comparative example 3 | 8.36 |
3. subjective appreciation：
Valuation officer feels product with all kinds of Score indexes of clear, accurate stepping qualitative description sample such as (table 3)
Official's evaluation.The sensory evaluation scores of averagely each sample are calculated according to the score of every valuation officer.
Table 3：Semen sojae atricolor polypeptide wine sensory evaluation criteria
Note：Score value is represented in bracket.
The sensory evaluation scores measurement result of semen sojae atricolor polypeptide wine prepared by embodiment 1, comparative example 1-3 is shown in Table 4.
Table 4：The sensory evaluation scores measurement result of semen sojae atricolor polypeptide wine
| Product | Subjective appreciation |
| Embodiment 1 | 88.6 |
| Comparative example 1 | 64.5 |
| Comparative example 2 | 71.2 |
| Comparative example 3 | 74.7 |
As can be seen from Table 4, its color of semen sojae atricolor polypeptide wine, state, mouthfeel, taste and smell prepared by the present invention are superior to
Polypeptide wine prepared by comparative example 1- comparative example 3.
Test example 2：The inoxidizability of semen sojae atricolor polypeptide wine measures
One, test method：
(1) DPPH Scavenging activity measures
Respectively in the certain density sample solution 4mL that takes for the 1st to the 9th day of fermentation, 1mL dehydrated alcohol (analysis is added
It is pure) prepare DPPH solution and make the final concentration of 0.1mmol/L of DPPH.Firmly then shaking mixes, and is placed on quiet in darkroom
30min is set, measures absorbance under 517nm.DPPH clearance rate is calculated as follows.
DPPH clearance rate %=100 × [1- (Ax-Axo)/Ao]
In formula：Ax is the absorbance being added after DPPH and sample solution；Axo is the absorbance of sample solution；Ao is to steam
The absorbance of distilled water and DPPH.
(2)O2 -Clearance rate measurement
Respectively at the 1st to the 9th day of fermentation, measured using assay NBT photoreduction.Take 50mmol/LTris-HCl slow
Fliud flushing (pH8.2) 4.5mL is placed in 25 DEG C of water-baths heat preservation 20min, be separately added into 0.4mL 25mmol/L pyrogallol solution and
1mL sample solution stops reaction, surveys at 299nm after mixing in 25 DEG C of water-bath 5min, the 8mmol/L HCl that 1mL is added
Determine absorbance A x, blank control group replaces sample with same volume distilled water.Each sample does 3 Duplicate Samples, is averaged, presses
Following formula calculates O2 -Clearance rate.
O2 -Clearance rate/%=100 × (Ao-Ax)/Ao
In formula：Ao is placebo solution absorbance；Ax is the absorbance of sample solution.
(3) reducing power measures
Respectively at the 1st to the 9th day of fermentation, using potassium ferricyanide method.1mL polypeptide wine solution is taken, is separately added into 2.5mL's
The phosphate buffer of 0.2mol/L1% potassium ferricyanide solution and pH6.6 are uniformly mixed it, and mixed liquor adds in 50 DEG C of water-baths
After hot 20min, adds 2.5mL10% trichloroacetic acid and terminate reaction.This mixed solution 2.5mL is drawn, 0.5mL 0.1% is added
Ferric trichloride, after mixing place 30min after in 700nm at measurement absorbance.
Two, test result：
The inoxidizability measurement result of semen sojae atricolor polypeptide wine prepared by embodiment 1, comparative example 1-3 is shown in Table 5.
Table 5：The inoxidizability measurement result of semen sojae atricolor polypeptide wine
| Product | DPPH clearance rate | O2 -Clearance rate | Reducing power |
| Embodiment 1 | 78.9% | 64.6% | 1.876 |
| Comparative example 1 | 56.4% | 45.3% | 1.152 |
| Comparative example 2 | 52.3% | 41.6% | 1.082 |
| Comparative example 3 | 54.5% | 42.1% | 1.174 |
| Comparative example 4 | 60.3% | 51.2% | 1.204 |
| Comparative example 5 | 58.6% | 48.7% | 1.195 |
As can be seen from Table 5, compared with the semen sojae atricolor polypeptide wine of other methods preparation, semen sojae atricolor polypeptide wine prepared by the present invention
Oxidation resistance is significantly improved.It is found through concertedness analysis, each raw material component has association in fermentation substrate of the invention
Same synergistic effect.
The foregoing is merely preferred embodiment of the present application, are not intended to limit this application, for the skill of this field
For art personnel, various changes and changes are possible in this application.Within the spirit and principles of this application, made any to repair
Change, equivalent replacement, improvement etc., should be included within the scope of protection of this application.
Claims (10)
1. a kind of preparation method of semen sojae atricolor polypeptide wine, which is characterized in that include the following steps：
(1) semen sojae atricolor albumen is digested, obtains semen sojae atricolor polypeptide liquid；
(2) fructus arctii is thinly sliced, is placed in the aqueous solution containing sodium citrate and impregnates 10-15min, it is dry, after obtaining pretreatment
Fructus arctii sample；The ethanol solution that 25-30 times of weight is added into pretreated fructus arctii sample carries out homogenate extraction, the second
The volumetric concentration of alcoholic solution is 70-80%, and extraction voltage is 100V, and extraction time 45-55s, extraction time is 2 times, is obtained
Fructus arctii extracting solution；By fructus arctii extracting solution microwave drying, burdock extract is obtained；
(3) ginkgo leaf crushed after being dried is added the water of 6-8 times of weight, extracts 1-3h at 80-100 DEG C at 80-100 mesh fine powder,
Filtering discards filter residue, and filtrate is concentrated and dried, and obtains ginkgo biloba extract；
(4) semen sojae atricolor polypeptide liquid, burdock extract and ginkgo biloba extract are mixed, adds sucrose, anhydrous sodium sulfite and water, goes out
Bacterium, as fermentation substrate；Adjusting the semen sojae atricolor content of peptides in fermentation substrate is 1.0%, and burdock extract content is 2.0-
4.0%, ginkgo biloba extract content is 1.0-3.0%, and cane sugar content 12-16%, anhydrous sodium sulfite content is 0.4-
0.6%；
(5) saccharomycete is accessed into fermentation substrate, is fermented 9 days at 26 DEG C；
(6) fermentation liquid is clarified using the method for centrifugation to get semen sojae atricolor polypeptide wine is arrived.
2. preparation method according to claim 1, which is characterized in that in step (1), the specific method of the enzymatic hydrolysis is：
Alcalase alkali protease is first used, semen sojae atricolor albumen is hydrolyzed under conditions of temperature is 60 DEG C, pH is 8.5；It is added after 30min
Flavourzyme flavor protease digests 30min under conditions of temperature is 50 DEG C, pH is 7.0；It adds later
Neutrase neutral proteinase digests 1h under conditions of temperature is 50 DEG C.
3. preparation method according to claim 1, which is characterized in that in step (2), fructus arctii is cut into a thickness of 0.3-
The thin slice of 0.5cm.
4. preparation method according to claim 1, which is characterized in that in step (2), lemon in the aqueous solution containing sodium citrate
The mass concentration of lemon acid sodium is 2-4%.
5. preparation method according to claim 1, which is characterized in that in step (2), the microwave drying controls vacuum pressure
Power is -0.1-0.05MPa, and drying temperature is 40-50 DEG C.
6. preparation method according to claim 1, which is characterized in that in step (4), the semen sojae atricolor adjusted in fermentation substrate is more
Peptide content is 1.0%, and burdock extract content is 4.0%, and ginkgo biloba extract content is 2.0%, and cane sugar content 16% is anhydrous
Sodium sulfite content is 0.5%.
7. preparation method according to claim 1, which is characterized in that in step (5), the access amount of the saccharomycete is hair
The 1.2 ‰ of ferment matrix weight.
8. preparation method according to claim 1 or claim 7, which is characterized in that in step (5), the saccharomycete is Angel ferment
The Angel Yeast of female limited liability company's production.
9. preparation method according to claim 1, which is characterized in that in step (6), centrifugal rotational speed 3500r/min, from
The heart time is 10min.
10. the semen sojae atricolor polypeptide wine that the described in any item methods of claim 1-9 are prepared.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112501227A (en) * | 2020-12-11 | 2021-03-16 | 山西娇禾生物科技有限公司 | Method for producing black bean polypeptide protein powder by biological fermentation |
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| CN1467285A (en) * | 2002-07-08 | 2004-01-14 | *** | Burdock and ginkgo health wine |
| KR20040039207A (en) * | 2004-02-17 | 2004-05-10 | 신명식 | kum cong ju |
| CN101230313A (en) * | 2007-01-24 | 2008-07-30 | 西北农林科技大学 | Walnut polypeptide wine and method for making same |
| CN102443516A (en) * | 2011-12-26 | 2012-05-09 | 青海省农林科学院 | Preparation method of broad bean polypeptide wine |
| CN105349307A (en) * | 2015-11-11 | 2016-02-24 | 广西辰昱生物科技有限公司 | Black soya bean wine and making method thereof |
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2018
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1467285A (en) * | 2002-07-08 | 2004-01-14 | *** | Burdock and ginkgo health wine |
| KR20040039207A (en) * | 2004-02-17 | 2004-05-10 | 신명식 | kum cong ju |
| CN101230313A (en) * | 2007-01-24 | 2008-07-30 | 西北农林科技大学 | Walnut polypeptide wine and method for making same |
| CN102443516A (en) * | 2011-12-26 | 2012-05-09 | 青海省农林科学院 | Preparation method of broad bean polypeptide wine |
| CN105349307A (en) * | 2015-11-11 | 2016-02-24 | 广西辰昱生物科技有限公司 | Black soya bean wine and making method thereof |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112501227A (en) * | 2020-12-11 | 2021-03-16 | 山西娇禾生物科技有限公司 | Method for producing black bean polypeptide protein powder by biological fermentation |
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Effective date of registration: 20221025 Address after: 402660 No. 269-2, Datong Street, Zitong Town, Tongnan District, Chongqing Patentee after: Wang Liangjun Address before: 221018 Lishui Road, Yunlong District, Xuzhou, Jiangsu 2 Patentee before: XUZHOU University OF TECHNOLOGY |