CN103589766A - Method for simultaneously extracting soybean peptide and soybean oligosaccharide from aqueous phase produced through aqueous enzymatic method - Google Patents
Method for simultaneously extracting soybean peptide and soybean oligosaccharide from aqueous phase produced through aqueous enzymatic method Download PDFInfo
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Abstract
The invention discloses a method for simultaneously extracting soybean peptide and soybean oligosaccharide from an aqueous phase produced through the aqueous enzymatic method and belongs to the technology of gain and oil bioprocessing. The method comprises the following steps: (1) soybeans are extruded and puffed, and then mixed with water for enzymolysis, and centrifugal separation is performed after enzymolysis, so that free oil, an emulsion, a hydrolysate and residues are obtained; (2) the emulsion, the hydrolysate and the residues are subjected to ultrasonic treatment to obtain a mixed liquor, two types of alkaline protease are added into the mixed liquor for step-by-step enzymolysis, centrifugal separation is performed after enzymolysis, so that free oil, an aqueous phase waste liquor and residues are obtained; (3) the acid deposition is performed on the aqueous phase waste liquor and then centrifugal separation is performed on the aqueous phase waste liquor to obtain sediment and aqueous phase mixture, and the sediment is subjected to vacuum concentration and spray drying to obtain soybean peptide; nanofiltration and ethanol precipitation are performed on the aqueous phase mixture to obtain sediment, and the sediment is subjected to decolorization and desalination treatment, and then subjected to vacuum concentration and spray drying to obtain the soybean oligosaccharide. According to the method, the aqueous phase mixed system formed during oil production through the aqueous enzymatic method is fully utilized, so that the waste is reduced, and the soybean peptide and the soybean oligosaccharide can be simultaneously obtained; the method has an excellent application prospect.
Description
Technical field
The invention belongs to grain and oil biological processing technology, relate generally to a kind of from aqueous enzymatic method water the method for simultaneous extraction soybean polypeptide and soybean oligosaccharide.
Background technology
It is on the basis of Mechanical Crushing that aqueous enzymatic method is carried oil tech, adopt enzyme to act on oil plant, make grease be easy to discharge from oil plant, utilize the avidity difference of non-oil component (protein and carbohydrate) to oil and water, utilize profit proportion different and oil is separated with non-oil component simultaneously.It carries oil tech as a kind of emerging " safety, green, environmental protection ", is more and more subject to lot of domestic and international scholar's attention.Can reclaim oil plant efficiently when extracting grease in, other value component (protein and carbohydrate etc.) ，Bei grease scientific circles are called " a kind of complete utilization technology of oil resource ".The grease of aqueous enzymatic extraction has good quality, and because enzymolysis carries out in water, functional substance in soybean, for example soybean isoflavones, soybean saponin, soybean peptides, soybean oligosaccharide, soybean dietary fibre isoreactivity material are all retained in water, because aqueous enzymatic method is put forward oil tech operational condition gentleness, the trace nutrient in raw material can be farthest retained, if its functional ingredient is separated, its added value of product can be improved.
Soybean peptides is that average peptide chain length is the small peptide (take 2-3 low molecular peptide as main) of 2-10, also contain the compositions such as a small amount of total free aminoacids, inorganic salt and carbohydrate, relative molecular mass take lower than 1000Da be main body, main relative molecular mass is distributed within the scope of 300Da-700Da.It is almost completely identical with soy-protein that soybean peptide ammino acid forms, the good and rich content of the balance of indispensable amino acid.Research finds that many small molecular weight titaniums, when through human body alimentary canal, are not hydrolyzed the utilization that can directly be absorbed by the body, and have multiple physiologically active, as hypotensive, oxidation-resistance, reduction cholesterol level etc.; Soybean oligosaccharide is the general name of soluble sugar in soybean.Mainly comprise sucrose, raffinose and stachyose, wherein monose and sucrose account for 65%, and raffinose accounts for 5%-7%, and stachyose accounts for 30%-32%.Raffinose and stachyose are functional oligose, have the effect of propagation bifidus bacillus.
Extruding and puffing technology is the high temperature, short time working method in a kind of modern times.Soybean is subject to mixing, compresses, mediates and shears in extruder, plastified and boiling, thereby to material component and microtexture generation effect, this directly has influence on following process effect through comprehensive actions such as moisture, temperature, pressure and mechanical shearings.In addition, when soybean passes through extrusion die orifice in a flash, high pressure sharply discharges, soybean moisture evaporates rapidly, make soybean by the inner tissue looseness's body that produces many tiny holes of expanded one-tenth, histocyte is damaged, thereby is conducive to the release of effective constituent (as grease, protein, isoflavones, saponin(e etc.) in cell.In addition, the extrusion process of high temperature, short time makes effective constituent in soybean be subject to the degree of heat collapse to be reduced to lower-most point.Therefore, in the soybean that extruding puffing is crossed, its effective constituent not only content loss is few, and is easily extracted.
Ultrasonic wave has judder and cavitation effect, and cavitation effect cell membrane plays destruction, can accelerate release and the stripping of effective constituent, and ultrasonic wave is constantly vibrated extracting solution, contributes to solute diffusion, thereby shortens extraction time, improves efficiency of pcr product.Therefore, ultrasonic supplementary means is widely used in natural product extraction research.
So far, the processing method that there is no a kind of technique simultaneous extraction soybean oil, soybean peptides and soybean oligosaccharide from aqueous enzymatic method water, and first extruding puffing-ultrasonication-substep highland barley seedlings by enzymolysis-membrane technology-resin absorption technology is combined, the effective constituent in simultaneous extraction soybean.Extruding puffing, ultrasonication combine, and have effectively destroyed the structure of cell, thereby are conducive to the release of effective constituent in cell, improve efficiency of pcr product; Utilize Sumizyme MP substep enzymolysis, reach breakdown of emulsion and the object of producing soybean peptides simultaneously; Utilize membrane technique and polymeric adsorbent technology to combine, soybean oligosaccharide is refined, obtain the soybean oligosaccharide product that purity is high.This technique is fully used the formed water mixed system of aqueous enzymatic method liquefaction, cut the waste, reduce costs, can obtain continuously 3 kinds of products, can be used as the raw material of protective foods and pharmaceuticals, and can carry out large-scale industrial production, there is significant economic benefit, environmental benefit and social benefit.
Summary of the invention
Technical problem to be solved by this invention is to overcome the deficiencies in the prior art, provide a kind of from aqueous enzymatic method water the method for simultaneous extraction soybean polypeptide and soybean oligosaccharide, reach the object of carrying high product yield, increasing added value.
Technical problem to be solved by this invention is achieved through the following technical solutions:
A kind of from aqueous enzymatic method water the method for simultaneous extraction soybean polypeptide and soybean oligosaccharide, the method comprises the following steps: (1) carries out expelling-expansion pretreatment after soybean cleaning is pulverized, obtain soybean extruding-puffing thing, after extruding puffing thing is mixed with water, carry out enzymolysis and obtain enzymolysis solution, enzymolysis solution centrifugation obtains free oil, milk sap, hydrolyzed solution and residue, (2) by milk sap, hydrolyzed solution and residue carry out supersound process and obtain mixed solution, described ultrasonic power is 250-450W, ultrasonic time is 10-30s, ultrasonic temperature is 60-80 ℃, first in mixed solution, add 2709 Sumizyme MPs to carry out enzymolysis, described enzyme concentration is 2%, hydrolysis temperature is 55 ℃, enzymolysis time is 1.5h, enzymolysis pH is 8.5, in mixed solution, add Sumizyme MP (Denmark novo company) to carry out enzymolysis again, described enzyme concentration is 3%, hydrolysis temperature is 55 ℃, enzymolysis time is 1.5h, enzymolysis pH is 9.0, after enzymolysis, carry out centrifugation and obtain free oil, water waste liquid and residue, (3) in water waste liquid, add acid for adjusting pH to 4.5, carry out the heavy processing of acid, then centrifugation is precipitated and aqueous mixture, precipitation is through vacuum concentration, spraying is dried to obtain soybean peptides, aqueous mixture is at 35 ℃, under 0.85MPa, carry out nanofiltration and must see through liquid, its molecular weight cut-off is 200-1000Da, seeing through liquid carries out alcohol precipitation and processes and to obtain precipitation, described alcohol concn is 80-100%, the alcohol steep time is 70-110min, alcohol steep temperature is 40-60 ℃, by precipitation by the desalination of decolouring of two kinds of series connection polymeric adsorbents, two kinds of described series connection resins are DM130 resin and D290 resin, adsorption temp is 55 ℃, absorption flow velocity is 35m
3/ (m
3h), decolouring desalination is by vacuum concentration, the dry soybean oligosaccharide that obtains of spraying.
Described supersound process preferred parameter is: ultrasonic power 297.19W, ultrasonic time 21.56s, 74.42 ℃ of ultrasonic temperature.
Described alcohol precipitation is processed preferred parameter: alcohol concn 93.15%, alcohol steep time 89.66min, 49.11 ℃ of alcohol steep temperature.
The invention belongs to the extraction processing technology of Vegetable oil lipoprotein, utilize physics and chemistry and biotechnology to combine, simultaneous extraction soybean oil, soybean peptides and soybean oligosaccharide effective constituent from aqueous enzymatic method water, adopt extruding and puffing technology-ultrasonic technology to combine, effectively destroyed the structure of cell, thereby be conducive to the release of effective constituent in cell, improve efficiency of pcr product; Utilize two kinds of Sumizyme MP substep enzymolysis, can not only be hydrolyzed the protein in water, can also destroy the lipoprotein membrane that is wrapped in oil droplets that oil plant forms in the processes such as defibrination, reduce the stability of milk sap, thereby improve free oil yield, when reaching breakdown of emulsion, obtain soybean peptides; Utilize membrane technique and polymeric adsorbent technology to combine, soybean oligosaccharide is refined, obtain the soybean oligosaccharide product that purity is high.The needed equipment of this technique is simple, operational safety, can synchronously obtain 3 kinds of products, can carry out serialization and produce multiple product, has significant economic benefit, environmental benefit and social benefit.
The present invention combines extruding puffing-ultrasonication-substep highland barley seedlings by enzymolysis-membrane technology-resin absorption technology first, the effective constituent in simultaneous extraction soybean, and soybean polypeptide yield can reach 94.46%, and soybean oligosaccharide yield can reach 67.87%.
Accompanying drawing explanation
Fig. 1 process route chart of the present invention;
The impact of Fig. 2 ultrasonic power on soybean polypeptide yield and soybean oligosaccharide yield;
The impact of Fig. 3 ultrasonic time on soybean polypeptide yield and soybean oligosaccharide yield;
The impact of the ultrasonic temperature of Fig. 4 on soybean polypeptide yield and soybean oligosaccharide yield;
Fig. 5
y=f (x 1 , x 2 )response surface;
Fig. 6
y=f (x 1 , x 3 )response surface;
Fig. 7
y=f (x 2 , x 3 )response surface;
The impact of Fig. 8 alcohol concn on soybean oligosaccharide yield;
The impact of Fig. 9 alcohol steep time on soybean oligosaccharide yield;
The impact of Figure 10 alcohol steep temperature on soybean oligosaccharide extraction yield;
Figure 11
y=f (x 1 , x 2 )response surface;
Figure 12
y=f (x 1 , x 3 )response surface;
Figure 13
y=f (x 2 , x 3 )response surface.
Embodiment
Below in conjunction with accompanying drawing, the specific embodiment of the invention is described in detail:
A kind of from aqueous enzymatic method water the method for simultaneous extraction soybean polypeptide and soybean oligosaccharide, the method comprises the following steps: (1) carries out expelling-expansion pretreatment after soybean cleaning is pulverized, obtain soybean extruding-puffing thing, after extruding puffing thing is mixed with water, carry out enzymolysis and obtain enzymolysis solution, enzymolysis solution centrifugation obtains free oil, milk sap, hydrolyzed solution and residue, (2) by milk sap, hydrolyzed solution and residue carry out supersound process and obtain mixed solution, described ultrasonic power is 250-450W, ultrasonic time is 10-30s, ultrasonic temperature is 60-80 ℃, first in mixed solution, add 2709 Sumizyme MPs to carry out enzymolysis, described enzyme concentration is 2%, hydrolysis temperature is 55 ℃, enzymolysis time is 1.5h, enzymolysis pH is 8.5, in mixed solution, add Sumizyme MP (Denmark novo company) to carry out enzymolysis again, described enzyme concentration is 3%, hydrolysis temperature is 55 ℃, enzymolysis time is 1.5h, enzymolysis pH is 9.0, after enzymolysis, carry out centrifugation and obtain free oil, water waste liquid and residue, (3) in water waste liquid, add acid for adjusting pH to 4.5, carry out the heavy processing of acid, then centrifugation is precipitated and aqueous mixture, precipitation is through vacuum concentration, spraying is dried to obtain soybean peptides, aqueous mixture is at 35 ℃, under 0.85MPa, carry out nanofiltration and must see through liquid, its molecular weight cut-off is 200-1000Da, seeing through liquid carries out alcohol precipitation and processes and to obtain precipitation, described alcohol concn is 80-100%, the alcohol steep time is 70-110min, alcohol steep temperature is 40-60 ℃, by precipitation by the desalination of decolouring of two kinds of series connection polymeric adsorbents, two kinds of described series connection resins are DM130 resin and D290 resin, adsorption temp is 55 ℃, absorption flow velocity is 35m
3/ (m
3h), decolouring desalination is by vacuum concentration, the dry soybean oligosaccharide that obtains of spraying.
Described supersound process preferred parameter is: ultrasonic power 297.19W, ultrasonic time 21.56s, 74.42 ℃ of ultrasonic temperature.
Described alcohol precipitation is processed preferred parameter: alcohol concn 93.15%, alcohol steep time 89.66min, 49.11 ℃ of alcohol steep temperature.
The screening experiment of embodiment 1 supersonic treatment conditional parameter
1 materials and methods
1.1 materials, reagent
1.2 key instrument equipment
1.3 experimental technique
1.3.1 the mensuration of soybean components
The mensuration of moisture: GB304-87 are measured; The mensuration of crude fat: in GB5512-85, Soxhlet extraction process is measured; The mensuration of crude protein: GB6432-94 standard method is carried out; Determination of ash: GB5009.4-85.
1.3.2 technical process (see figure 1)
1.3.3 soybean polypeptide yield determination: Kjeldahl determination method.
In formula
---soluble nitrogen in 10%TCA, mg
1.3.4 the mensuration of soybean oligosaccharide extraction yield:
2 results and discussion
2.1 impacts of supersonic treatment list factor condition on soybean polypeptide yield and soybean oligosaccharide yield
2.1.1 the impact of ultrasonic power on soybean polypeptide yield and soybean oligosaccharide yield
At ultrasonic time, be 20s, ultrasonic temperature is under 70 ℃ of conditions, investigates the impact of ultrasonic power on polypeptide yield and soybean oligosaccharide yield, the results are shown in Figure 2.As shown in Figure 2, along with the increase of ultrasonic power, polypeptide yield and soybean oligosaccharide yield significantly raise; But when ultrasonic power reaches 300W, along with the increase of ultrasonic power, polypeptide yield and soybean oligosaccharide yield decline on the contrary, therefore, in response surface test design below, ultrasonic power level is selected 280-320W.
2.1.2 the impact of ultrasonic time on soybean polypeptide yield and soybean oligosaccharide yield
At ultrasonic power, be 300W, ultrasonic temperature is under 70 ℃ of conditions, investigates the impact of ultrasonic time on polypeptide yield and soybean oligosaccharide yield, the results are shown in Figure 3.By Fig. 3 result, can be found out, when ultrasonic time is less than 20s, increase along with ultrasonic time, soybean oligosaccharide yield is in rising trend, when ultrasonic time reaches 20s, soybean oligosaccharide yield reaches maximum value, and while being greater than 20s, along with the increase of ultrasonic time, soybean oligosaccharide yield is on a declining curve on the contrary; When ultrasonic time is less than 25s, increase along with ultrasonic time, polypeptide yield is in rising trend, when ultrasonic time reaches 25s, polypeptide yield reaches maximum value, and while being greater than 25s, along with the increase of ultrasonic time, polypeptide yield is on a declining curve on the contrary, so ultrasonic time test design level is selected 18-26s in response surface experimental design below.
2.1.3 the impact of ultrasonic temperature on soybean polypeptide yield and soybean oligosaccharide yield
At ultrasonic power, be 300W, ultrasonic time is under 20s condition, investigates the impact of ultrasonic temperature on polypeptide yield and soybean oligosaccharide yield, the results are shown in Figure 4.As shown in Figure 4, when ultrasonic temperature is less than 70 ℃, rising along with ultrasonic temperature, polypeptide yield and soybean oligosaccharide yield are in rising trend, when ultrasonic temperature reaches 70 ℃, polypeptide yield and soybean oligosaccharide yield reach maximum value, and while being greater than 70 ℃, along with the increase of ultrasonic temperature, polypeptide yield and soybean oligosaccharide yield are on a declining curve on the contrary.Therefore, in response surface test design below, ultrasonic temperature is chosen as 60-80 ℃.
The response surface optimization Test of 2.2 supersonic treatments
2.2.1 experimental factor level code table is in Table 1-1.
On the basis of single factor research, choosing ultrasonic power, ultrasonic time, 3 factors of ultrasonic temperature is independent variable(s), take soybean polypeptide yield as response value, according to center combination principle of design, and the experiment of design response surface analysis, its level of factor coding schedule is in Table 1-1.
Table 1-1 level of factor coding schedule
2.2.2 response surface test arrangement and test-results
This test application responds face optimized method carries out process optimization.With x
1, x
2, x
3for independent variable(s), take protein isolate extraction yield as response value Y, response surface testing program and the results are shown in Table 1-2.Test number 1-14 is that analytical test 15-20 is 6 center tests, in order to assess experimental error.
Table 1-2 response surface testing program and test-results
2.2.3 the response surface interpretation of result of supersonic treatment parameter to polypeptide yield
By statistical analysis software DesignExpert7.1.1, carry out data analysis, set up Quadratic response surface regression model as follows:
Y=93.37+0.51x
1+0.73x
2+1.69x
3-3.32x
1x
2-5.07x
1x
3-4.64x
2x
3-3.98x
1 2-3.06x
2 2-2.04x
3 2
Regression analysis and the results of analysis of variance are in Table 1-3, and significant response surface analysis is shown in Fig. 5-Fig. 7 mutually alternately.
Table 1-3 returns and the results of analysis of variance
Note: by analysis, the relative coefficient (R of total regression
2) be 99.17%, the coefficient of determination (R
2 adj) be 98.43%
From showing 1-3, the linear relationship between equation dependent variable and independent variable(s) is obvious, and this model returns extremely significantly (p<0.0001), loses plan item not remarkable, and this model R
2=99.17%, R
2 adj=98.43%, illustrate that this model is good with test matching, between independent variable(s) and response value, linear relationship is remarkable, can infer for the theory of this reaction.By F check, can obtain factor contribution rate is: x
3>x
2>x
1, i.e. ultrasonic temperature > ultrasonic time > ultrasonic power.
Application responds face optimizing analytical procedure is analyzed regression model, and when ultrasonic power is 297.19W, ultrasonic time is 21.56s, and ultrasonic temperature is 74.42 ℃, and response surface has optimum value 94.4624 ± 0.39%.
2.2.4 proof test and simultaneous test
The top condition of trying to achieve in response surface analysis method at response surface analysis: ultrasonic power is 297.19W, ultrasonic time is 21.56s, ultrasonic temperature is under 74.42 ℃ of conditions, carry out 3 parallel tests, the mean value that obtains 3 parallel test polypeptide yield is 94.63%, illustrates that the experimental value of response value and regression equation predictor are identical good.
2.2.5 test brief summary
The supersonic treatment parameter of utilizing Response surface methodology to extract soybean polypeptide from aqueous enzymatic method water is optimized.Having set up corresponding mathematical model is that later pilot scale and suitability for industrialized production provided fundamental basis, and has obtained optimum supersonic treatment parameter: ultrasonic power is 297.19W, and ultrasonic time is 21.56s, and ultrasonic temperature is 74.42 ℃.Through checking with simultaneous test known under optimum supersonic treatment Parameter Conditions soybean polypeptide yield can reach 94.46% left and right.
The screening experiment of embodiment 2 alcohol precipitation treatment process condition parameters
1 materials and methods
1.1 materials, reagent
1.2 key instrument equipment
| Plant and instrument | Producer |
| Whizzer | Beijing Medical Centrifugal Machine Factory |
| RE-52A Rotary Evaporators | Shanghai Yarong Biochemical Instrument Plant |
1.3 experimental technique
1.3.1 technical process (see figure 1)
1.3.2 the mensuration of soybean oligosaccharide extraction yield:
2 results and discussion
2.1 alcohol precipitations are processed the impact on soybean oligosaccharide yield
2.1.1 the impact of alcohol concn on soybean oligosaccharide yield
At alcohol steep time 90min, under 40 ℃ of conditions of alcohol steep temperature, investigate the impact of alcohol concn on soybean oligosaccharide yield, the results are shown in Figure 8.As shown in Figure 8, along with the increase of alcohol concn, soybean oligosaccharide yield significantly raises; When alcohol concn reaches 95%, soybean oligosaccharide yield reaches maximum value, and while being greater than 95%, along with the increase of alcohol concn, soybean oligosaccharide yield is on a declining curve on the contrary, and therefore, in response surface test design below, alcohol concn is chosen as 90-98%.
2.1.2 the impact of alcohol steep time on soybean oligosaccharide yield
At alcohol concn, be 95%, alcohol steep temperature is under 40 ℃ of conditions, investigates the impact of alcohol steep time on soybean oligosaccharide yield, the results are shown in Figure 9.As shown in Figure 9, when the alcohol steep time reaches 90min, soybean oligosaccharide yield all reaches maximum value, therefore in response surface test design below alcohol steep selection of time at 80-100min.
2.1.3 the impact of alcohol steep temperature on soybean oligosaccharide yield
At alcohol concn, be 95%, under alcohol steep time 90min condition, investigate the impact of alcohol steep temperature on soybean oligosaccharide yield, the results are shown in Figure 10.As shown in Figure 10, along with the increase of alcohol steep temperature, soybean oligosaccharide yield significantly raises; But when alcohol steep temperature reaches 45 ℃, along with the increase of alcohol steep temperature, soybean oligosaccharide yield declines on the contrary, therefore, in response surface test design below, alcohol steep temperature levels is selected 35-55 ℃.
The response surface optimization Test of 2.2 alcohol precipitation treatment process
2.2.1 experimental factor level code table is in Table 1-1.
On the basis of single factor research, choosing alcohol concn, alcohol steep time, 3 factors of alcohol steep temperature is independent variable(s), take soybean oligosaccharide yield as response value, according to center combination principle of design, the experiment of design response surface analysis, its level of factor coding schedule is in Table 1-1.
Table 1-1 level of factor coding schedule
2.2.2 response surface test arrangement and test-results
This test application responds face optimized method carries out process optimization.With x
1, x
2, x
3for independent variable(s), take soybean oligosaccharide yield as response value Y, response surface testing program and the results are shown in Table 1-2.Test number 1-14 is that analytical test 15-20 is 6 center tests, in order to assess experimental error.
Table 1-2 response surface testing program and test-results
2.2.3 the response surface interpretation of result of alcohol precipitation treatment process parameter to soybean oligosaccharide yield
By statistical analysis software DesignExpert7.1.1, carry out data analysis, set up Quadratic response surface regression model as follows:
Y=66.97+0.059x
1+0.96x
2+1.28x
3-1.52x
1x
2-4.02x
1x
3-3.21x
2x
3-2.97x
1 2-2.30x
2 2-1.57x
3 2
Regression analysis and the results of analysis of variance are in Table 1-3, and significant response surface analysis is shown in Figure 11-Figure 13 mutually alternately.
Table 1-3 returns and the results of analysis of variance
Note: by analysis, the relative coefficient (R of total regression
2) be 98.82%, the coefficient of determination (R
2 adj) be 97.75%
From showing 1-3, the linear relationship between equation dependent variable and independent variable(s) is obvious, and this model returns extremely significantly (p<0.0001), loses plan item not remarkable, and this model R
2=98.82%, R
2 adj=97.75%, illustrate that this model is good with test matching, between independent variable(s) and response value, linear relationship is remarkable, can infer for the theory of this reaction.By F check, can obtain factor contribution rate is: x
3>x
2>x
1, i.e. alcohol steep temperature > alcohol steep time > alcohol concn.
Application responds face optimizing analytical procedure is analyzed regression model, and when alcohol concn is 93.15%, the alcohol steep time is 89.66min, and alcohol steep temperature is 49.11 ℃, and response surface has optimum value 67.8661 ± 0.34%.
2.2.4 proof test and simultaneous test
The top condition of trying to achieve in response surface analysis method at response surface analysis: alcohol concn is 93.15%, the alcohol steep time is 89.66min, alcohol steep temperature is under 49.11 ℃ of conditions, carry out 3 parallel tests, the mean value that obtains 3 parallel test polypeptide yield is 67.38%, illustrates that the experimental value of response value and regression equation predictor are identical good.
2.2.5 test brief summary
The supersonic treatment parameter of utilizing Response surface methodology to extract soybean polypeptide from aqueous enzymatic method water is optimized.Having set up corresponding mathematical model is that later pilot scale and suitability for industrialized production provided fundamental basis, and has obtained optimum supersonic treatment parameter: alcohol concn is 93.15%, and the alcohol steep time is 89.66min, and alcohol steep temperature is 49.11 ℃.Through checking with simultaneous test known under optimum supersonic treatment Parameter Conditions soybean polypeptide yield can reach 67.87% left and right.
Claims (3)
1. the method for simultaneous extraction soybean polypeptide and soybean oligosaccharide from an aqueous enzymatic method water, it is characterized in that the method comprises the following steps: (1) carries out expelling-expansion pretreatment after soybean cleaning is pulverized, obtain soybean extruding-puffing thing, after extruding puffing thing is mixed with water, carry out enzymolysis and obtain enzymolysis solution, enzymolysis solution centrifugation obtains free oil, milk sap, hydrolyzed solution and residue, (2) by milk sap, hydrolyzed solution and residue carry out supersound process and obtain mixed solution, described ultrasonic power is 250-450W, ultrasonic time is 10-30s, ultrasonic temperature is 60-80 ℃, first in mixed solution, add 2709 Sumizyme MPs to carry out enzymolysis, described enzyme concentration is 2%, hydrolysis temperature is 55 ℃, enzymolysis time is 1.5h, enzymolysis pH is 8.5, in mixed solution, add Sumizyme MP (Denmark novo company) to carry out enzymolysis again, described enzyme concentration is 3%, hydrolysis temperature is 55 ℃, enzymolysis time is 1.5h, enzymolysis pH is 9.0, after enzymolysis, carry out centrifugation and obtain free oil, water waste liquid and residue, (3) in water waste liquid, add acid for adjusting pH to 4.5, carry out the heavy processing of acid, then centrifugation is precipitated and aqueous mixture, precipitation is through vacuum concentration, spraying is dried to obtain soybean peptides, aqueous mixture is at 35 ℃, under 0.85MPa, carry out nanofiltration and must see through liquid, its molecular weight cut-off is 200-1000Da, seeing through liquid carries out alcohol precipitation and processes and to obtain precipitation, described alcohol concn is 80-100%, the alcohol steep time is 70-110min, alcohol steep temperature is 40-60 ℃, by precipitation by the desalination of decolouring of two kinds of series connection polymeric adsorbents, two kinds of described series connection resins are DM130 resin and D290 resin, adsorption temp is 55 ℃, absorption flow velocity is 35m
3/ (m
3h), decolouring desalination is by vacuum concentration, the dry soybean oligosaccharide that obtains of spraying.
According to claim 1 a kind of from aqueous enzymatic method water the method for simultaneous extraction soybean polypeptide and soybean oligosaccharide, it is characterized in that described supersound process preferred parameter is: ultrasonic power 297.19W, ultrasonic time 21.56s, 74.42 ℃ of ultrasonic temperature.
According to claim 1 a kind of from aqueous enzymatic method water the method for simultaneous extraction soybean polypeptide and soybean oligosaccharide, it is characterized in that described alcohol precipitation processing preferred parameter is: alcohol concn 93.15%, alcohol steep time 89.66min, 49.11 ℃ of alcohol steep temperature.
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| CN108635393A (en) * | 2018-07-17 | 2018-10-12 | 安徽天安生物科技股份有限公司 | A kind of anti-oxidant occrycetin injection and preparation method thereof |
| CN108949887A (en) * | 2018-09-04 | 2018-12-07 | 哈尔滨工业大学 | A kind of preparation method of the multi-functional incretin peptide of soybean |
| CN109206475A (en) * | 2018-09-17 | 2019-01-15 | 东北农业大学 | A method of it is digested using single step while extracting oil body and albumen |
| CN109734753A (en) * | 2018-11-27 | 2019-05-10 | 安徽博悦生物科技有限公司 | A kind of extracting method of soyabean oligosaccharides |
| CN109691553A (en) * | 2019-02-13 | 2019-04-30 | 黑龙江八一农垦大学 | A kind of imitative meat product and its processing method rich in soya-bean polypeptides |
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| CN111184219A (en) * | 2020-01-09 | 2020-05-22 | 东北农业大学 | Method for preparing polypeptide compound by combining tea polyphenol and polypeptide |
| CN113699197A (en) * | 2021-09-01 | 2021-11-26 | 吉林大学 | Process for extracting cyperus esculentus oligosaccharide by enzyme method |
| CN113699197B (en) * | 2021-09-01 | 2023-04-07 | 吉林大学 | Process for extracting cyperus esculentus oligosaccharide by enzyme method |
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