CN103564145A - Method for extracting high-solubility black soya bean protein - Google Patents
Method for extracting high-solubility black soya bean protein Download PDFInfo
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Abstract
The invention discloses a method for extracting a high-solubility black soya bean protein, which belongs to a plant protein processing technology. The method comprises the following steps: (1) after black soya beans are cleared and crushed, sieving the obtained object so as to obtain black soya bean flour, mixing the black soya bean flour with normal hexane so as to degrease the black soya bean flour, and after degreasing is completed, carrying out centrifugal separation on the obtained object so as to obtain degreased black soya bean flour; (2) mixing the degreased black soya bean flour with water so as to obtain mixed liquid, and adjusting the pH value of the mixed liquid; (3) carrying out ultrasonic processing on the mixed liquid; (4) carrying out pulsed electric field treatment on the mixed liquid subjected to ultrasonic processing; and (5) carrying out centrifugal separation on the mixed liquid subjected to pulsed electric field treatment so as to obtain liquid supernatant, adjusting the pH value of the liquid supernatant to 4.5, carrying out centrifugal separation on the liquid supernatant so as to obtain a precipitation, and carrying out repeated washing and centrifugal separation on the precipitation so as to obtain the high-solubility black soya bean protein. According to the invention, a high-solubility black soya bean protein can be obtained, required devices are simple and safe in operation, the extraction rate of the obtained black soya bean protein can reach 70.04%, and the solubility can be up to 62.29%.
Description
Technical field
The invention belongs to vegetable protein process technology, relate generally to a kind of extracting method of highly dissoluble black soya bean albumen.
Background technology
In recent years, along with " black-food " theoretical birth, the nutritive value of black soya bean and medical value are further familiar with, and the development of process technology has promoted the variation of black soya bean product, has increased the use value of black soya bean.Black soya bean albumen, as the typical component of nutritious black bean composition, is not only better than various plants albumen on protein content, and its amino acid composition, the property digested and assimilated are all comparable to animal protein simultaneously.Black soya bean contains rich in protein, and black soya bean protein content reaches as high as 49. 8%, higher by 24.5% than soya bean, even higher than meat, egg and milk, have the good reputation of " king of vegetable protein ".In addition, in black soya bean, be also rich in isoflavones, vitamin E, carotenoid, anthocyanidin and polyphenols.Black soya bean albumen not only content is high, and quality is good, and wherein the amino acid of protein forms similar to animal protein.In black soya bean protein, contain 18 seed amino acids, and essential amino acid accounts for the more than 40% of total amino acid amount, meets FAO/WHO balanced mode standard, there is good amino acid balance, being a kind of complete protein, therefore more easily digesting and assimilating, is a kind of ideal, potential high-quality protein resource.The black bean protein powder being made into meets the desirable Model of Nutrition of recommendation simultaneously very much, and from edible angle, black bean protein powder good mouthfeel, easy to carry, be consumers in general's health foods loved by all.But due to black soya bean protein conformation closely, highly folding, cause black soya bean protein structure compliance poor, the corresponding reduction of whole dissolubility, and then due to bad dissolving ambient influnence the performance of the functional characters such as emulsibility, foaming characteristic, greatly limited Industry Promotion and the range of application of black soya bean albumen.
Ultrasonic effect day in food industry is aobvious outstanding, as processing, and the aspects such as fresh-keeping and extraction.Supersonic modifying belongs to a kind of of physical modification method, compares its few, workable advantage that has accessory substance with chemical modification.The food processing process of same complete reproduction, now can by use ultrasonic wave within several seconds or a few minutes with the high repeatability that completes, not only can reduce processing cost, simplify the operation, and only consume fraction and just can provide the purity of higher final products.In addition, the treatment technology at protein can be considered to a kind of feasible technology for the mode of quality assurance and food security.Ultrasonic wave have heat effect, mechanism, cavitation and many second order effects as smashed, emulsification, diffusion etc., can effectively break cell boundaries layer, diffusion velocity is increased, promote the stripping of active ingredient in solid material, can improve significantly extraction efficiency.
Impulse electric field (Pulsed Electric Fields, PEF) can to liquid, semi-solid food products, process with higher electric-field intensity (10-50kV/cm), shorter pulse width (0-100 μ s) and higher pulse frequency (0-2000Hz), make microorganism membrane perforation, break, the structural deterioration of enzyme, reaches the object of the blunt enzyme of sterilization.The burst process time is short, and heat-energy losses is low, and food composition changes little, can preserve to greatest extent local flavor, color and luster, mouthfeel and the nutritive value of food.In recent years, impulse electric field was also more and more extensive in the research in albumen field.Research shows the polarization phenomena that PEF can inducible protein molecule, destroys the hydrophobic interaction power that maintains albumen space structure, and protein molecular is partly stretched, and hydrophobic grouping exposes, and functional character changes.External aspect high-pressure pulse electric, done many research, and great majority research is all the aspect that affects on food microorganisms and enzyme for high-pressure pulse electric, about high-pressure pulse electric on the impact research of food composition seldom.
At present, the method for extraction vegetable protein has cold-press, lixiviation process, the heavy method of the molten acid of alkali, the heavy method of acid, the extraction of alcohol method, aqueous enzymatic method and membrane separation technique etc.But these methods extraction times is long or extraction cost is higher, and this patent adopts ultrasonic pulse electric field auxiliary law to extract black soya bean albumen on the basis of the heavy method of the molten acid of alkali, significantly improves the recovery rate of black soya bean albumen.The method of ultrasonic pulse electric field assisted extraction albumen combines ultrasonic vibration with impulse electric field, make full use of the cavitation of ultrasonic vibration and impulse electric field effect, shortened extraction time, improved extraction efficiency, promote the lifting of black soya bean protein solubility simultaneously, in the technique of modern times extraction albumen, also almost there is no the research of the technique of this respect.
Summary of the invention
Technical problem to be solved by this invention is to overcome the deficiencies in the prior art, and a kind of extracting method of highly dissoluble black soya bean albumen is provided.
Technical problem to be solved by this invention is achieved through the following technical solutions:
A kind of extracting method of highly dissoluble black soya bean albumen, the method comprises the following steps: (1) is crossed 60 mesh sieves after black soya bean cleaning is pulverized and obtained black bean powder, black bean powder is mixed in 1:3 ratio with n-hexane, and at room temperature stir 2h and carry out degreasing, after degreasing, centrifugation obtains degreasing black bean powder; (2) degreasing black bean powder and water are mixed to get to mixed liquor, and the mass fraction that regulates the pH of mixed liquor, described black bean powder to account for water is 0.07-0.11, the pH of mixed liquor is 7-11; (3) mixed liquor is carried out to ultrasonic processing, described ultrasonic power is 100-500W, and ultrasonic time is 10-30min; (4) mixed liquor after ultrasonic processing is carried out to impulse electric field processing, described pulse width is 2-6 μ s, and sample flow rate is 40-80ml/min, and the burst length is 60-500 μ s, and impulse electric field strength is 10-40kV; (5) the mixed liquor centrifugal 40min under 10000r/min after impulse electric field processing is obtained to supernatant, supernatant pH is adjusted to 4.5, centrifugation is precipitated, and will precipitate washing and centrifugation repeatedly, obtains the black soya bean albumen of highly dissoluble.
The mass fraction that described preferred black bean powder accounts for water is 0.094, and preferably the pH of mixed liquor is 9.72.
Described ultrasonic processing preferred parameter is: ultrasonic power 378.47W, ultrasonic time 23.91min.
Described impulse electric field is processed preferred parameter: pulse width 4.3 μ s, sample flow rate 61ml/min, burst lengths 369 μ s, impulse electric field strength 30kV.
The invention belongs to the extraction process technology of vegetable protein, utilize physics and chemistry and biotechnology to combine, extract high extraction and have highly dissoluble black soya bean albumen concurrently, adopting ultrasonic wave to process combines with impulse electric field, because ultrasonic wave has heat effect, mechanism, cavitation and many second order effects are as smashed, emulsification, diffusion etc., can effectively break cell boundaries layer, diffusion velocity is increased, promote the stripping of protein in defatted soy flour, can improve significantly extraction efficiency, and at ultrasonic wave, process the enterprising stepping horizontal pulse electric field treatment in basis of degreasing black bean powder, black soya bean albumen is easier to be distributed to water at alkaline environment, the principle of utilizing protein to separate out in isoelectric point, by Protein Extraction out, recovery rate is improved, the processing of impulse electric field simultaneously, the polarization phenomena of protein molecular have been induced, destruction maintains the hydrophobic interaction power of albumen space structure, protein molecular is partly stretched, the dissolubility of black soya bean albumen is significantly improved, thereby prepare the black soya bean albumen of high extraction and highly dissoluble.This byproduct in process thing is few, workable, and needed equipment is simple, handling safety, with low cost, simplifies the operation, and only consumes fraction and just can provide the purity of higher final products.The black soya bean albumen no solvent residue of gained, quality and food security guarantee to some extent, the black soya bean protein solubility simultaneously obtaining is high, for the high value product of General Promotion black soya bean intensive processing technology, in-depth pollution-free food machining based conception provide intelligence support.
The present invention adopts ultrasonic wave and impulse electric field associated treatment alkalescence bean powder mixed liquor, then, by regulating pH to extract black soya bean albumen to the mode of albumen isoelectric point, can access the black soya bean albumen of high extraction, obtains having the black soya bean albumen of highly dissoluble simultaneously.
The present invention mainly has following technological advantage than the method for the heavy leach protein of the molten acid of traditional alkali:
(1) supersonic synergic impulse electric field is processed the recovery rate that can significantly increase black soya bean albumen;
(2) simple, the handling safety of equipment, cost are low, only consume fraction and just can provide the purity of higher final products;
(3) adopt the processing of impulse electric field, induced the polarization phenomena of protein molecular, destroy the hydrophobic interaction power that maintains albumen space structure, protein molecular is partly stretched, the dissolubility of black soya bean albumen is significantly improved.
Accompanying drawing explanation
Fig. 1 process route chart of the present invention;
The impact of Fig. 2 material percentage on black soya bean protein extracting ratio;
The impact of Fig. 3 pH on black soya bean protein extracting ratio;
The impact of Fig. 4 ultrasonic power on black soya bean protein extracting ratio;
The impact of Fig. 5 ultrasonic time on the impact of black soya bean protein extracting ratio;
Fig. 6 Y=f (x
1, x
2) response surface;
Fig. 7 Y=f (x
1, x
3) response surface;
Fig. 8 Y=f (x
1, x
4) response surface;
Fig. 9 Y=f (x
2, x
3) response surface;
Figure 10 Y=f (x
2, x
4) response surface;
Figure 11 Y=f (x
3, x
4) response surface;
The impact of Figure 12 pulse width on black soya bean protein extracting ratio;
The impact of Figure 13 sample flow rate on black soya bean protein extracting ratio;
The impact of Figure 14 burst length on black soya bean protein extracting ratio;
The impact of Figure 15 impulse electric field strength on black soya bean protein extracting ratio;
Figure 16 Y=f (x
1, x
2) response surface;
Figure 17 Y=f (x
1, x
3) response surface;
Figure 18 Y=f (x
1, x
4) response surface;
Figure 19 Y=f (x
2, x
3) response surface;
Figure 20 Y=f (x
2, x
4) response surface;
Figure 21 Y=f (x
3, x
4) response surface;
The calibration curve of the solubility of Figure 22 protein;
Figure 23 is ultrasonic-and impulse electric field processes the impact on black soya bean protein solubility.
The specific embodiment
Below in conjunction with accompanying drawing, the specific embodiment of the invention is described in detail:
A kind of extracting method of highly dissoluble black soya bean albumen, the method comprises the following steps: (1) is crossed 60 mesh sieves after black soya bean cleaning is pulverized and obtained black bean powder, black bean powder is mixed in 1:3 ratio with n-hexane, and at room temperature stir 2h and carry out degreasing, after degreasing, centrifugation obtains degreasing black bean powder; (2) degreasing black bean powder and water are mixed to get to mixed liquor, and the mass fraction that regulates the pH of mixed liquor, described black bean powder to account for water is 0.07-0.11, the pH of mixed liquor is 7-11; (3) mixed liquor is carried out to ultrasonic processing, described ultrasonic power is 100-500W, and ultrasonic time is 10-30min; (4) mixed liquor after ultrasonic processing is carried out to impulse electric field processing, described pulse width is 2-6 μ s, and sample flow rate is 40-80ml/min, and the burst length is 60-500 μ s, and impulse electric field strength is 10-40kV; (5) the mixed liquor centrifugal 40min under 10000r/min after impulse electric field processing is obtained to supernatant, supernatant pH is adjusted to 4.5, centrifugation is precipitated, and will precipitate washing and centrifugation repeatedly, obtains the black soya bean albumen of highly dissoluble.
The mass fraction that described preferred black bean powder accounts for water is 0.094, and preferably the pH of mixed liquor is 9.72.
Described ultrasonic processing preferred parameter is: ultrasonic power 378.47W, ultrasonic time 23.91min.
Described impulse electric field is processed preferred parameter: pulse width 4.3 μ s, sample flow rate 61ml/min, burst lengths 369 μ s, impulse electric field strength 30kV.
The screening experiment of experimental example 1 ultrasonic processing technique conditional parameter
1.1 materials and equipment
1.1.1 raw material and main agents
Black soya bean, it is pure that n-hexane, NaOH, hydrochloric acid etc. are commercially available analysis
1.1.2 instrument and equipment
| PHS-25 type acidometer | Shanghai great achievement instrument plant |
| Beater disintegrating machine | China Tianjin Stettlen Instrument Ltd. |
| Electric precise mixer | Jintan City, Jiangsu Province high honour instrument manufacturing Co., Ltd |
| Low speed centrifuge | Beijing Medical Centrifugal Machine Factory |
| Supercentrifuge | Germany Eppendorf company |
| Ultrasonic cell disrupte machine | NingBo XinZhi Biology Science Co., Ltd |
| Electric precise mixer | Jintan City, Jiangsu Province high honour instrument manufacturing Co., Ltd |
| Freeze dryer | U.S. SIM company |
1.3 experimental technique
1.3.1 the mensuration of black soya bean composition
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; Ash determination: GB5009.4-85.
1.3.2 black soya bean protein preparation method
Raw material black soya bean was pulverized 60 mesh sieves after peeling, fragmentation, then under 40 ℃ of conditions, adopted n-hexane extraction to prepare defatted soybean meal.The mass ratio of defatted soybean meal being pressed to 1:15 mixes with water, with 2mol/L NaOH adjust pH to 8.0, after stirring l.5h, by its suspension centrifugal 30min of 10000 * g under 4 ℃ of conditions, gets 2mol/L HCl adjust pH to 4.5 for supernatant.After standing under 4 ℃ of conditions the centrifugal 30min of 6000 * g, get albumen precipitation washing 2 times, finally get that precipitation is scattered in water and with 2mol/L NaOH adjust pH to 7.0.The centrifugal 30min of 10000 * g under 4 ℃ of conditions, removes a small amount of insoluble matter again, will after its protein solution freeze drying, pulverize and obtain Powdered black soya bean.
1.3.3 technological process (see figure 1)
1.3.4 computing formula
2 results and discussion
2.1 impacts of ultrasonic processing technique list factor condition on black soya bean protein extracting ratio
2.1.1 the impact of material percentage on black soya bean protein extracting ratio
Material percentage on the impact of recovery rate as Fig. 2.As shown in Figure 2, along with the reduction of solid-liquid ratio or the rising of concentration of substrate, the Protein Extraction then slow decreasing that takes the lead in raising, solid-liquid ratio is greater than 0.08 extraction rate of protein obviously to be increased, but when solid-liquid ratio is greater than 0.1, extraction rate of protein is on a declining curve, so solid-liquid ratio is selected 0.08-0.1 in response surface experimental design below.
2.1.2 the impact of pH on black soya bean protein extracting ratio
PH is shown in Fig. 3 to the impact of oil extracting rate.As shown in Figure 3, at solid-liquid ratio 1:8, ultrasonic time 20min, under ultrasonic power 300W condition, along with the increase of pH, protein extracting ratio increases gradually, when pH has higher value near 9-11, occurs, and recovery rate is the highest, and then, along with the increase of pH, recovery rate reduces gradually.So pH selects 9-11 in response surface experimental design below
2.1.3 the impact of ultrasonic power on black soya bean protein extracting ratio
Ultrasonic time is shown in Fig. 4 to the impact of protein extracting ratio.As shown in Figure 4, when the time is greater than 15min, recovery rate significantly rises, and intensity of variation is more obvious, and when the time is greater than 25min, recovery rate declines to a great extent.So ultrasonic time is selected 15-25min in response surface experimental design below.
2.1.4 the impact of ultrasonic time on black soya bean protein extracting ratio
Ultrasonic power is shown in Fig. 5 to the impact of protein extracting ratio.As shown in Figure 5, when power is greater than 200W, recovery rate rises, and intensity of variation is more obvious, and when power is greater than 400W, recovery rate declines.So ultrasonic power is selected 200-400W in response surface experimental design below.
The response surface Optimum Experiment of 2.2 ultrasonic processing technique
2.2.1 experimental factor level code table is in Table 1-1.
On the basis of single factor research, choose solid-liquid ratio, pH of mixed, ultrasound intensity, 4 factors of ultrasonic time are independent variable, take black soya bean protein extracting ratio as response, according to center combination design principle, the experiment of design response surface analysis, its factor level coding schedule is in Table 1-1.
Table 1-1 factor level coding schedule
2.2.2 response surface test arrangement and result of the test
This test application responds face optimization carries out process optimization.With x
1, x
2, x
3, x
4for independent variable, take protein isolate recovery rate as response Y, response surface testing program and the results are shown in Table 1-2.Tested number 1-24 is that analytical test 25-30 is 6 center tests, in order to assess experimental error.
Table 1-2 response surface testing program and result of the test
2.2.3 the response surface interpretation of result of ultrasonic processing technique parameter to protein isolate recovery rate
By statistical analysis software SAS9.2, carry out data analysis, set up Quadratic response surface regression model as follows:
Y=96.01-0.48x
1+1.87x
2+0.53x
3+2.40x
4-3.08x
1x
2-1.27x
1x
3+3.14x
1x
4-1.43x
2x
3+0.081?x
2?x
4-3.18?x
3?x
4-3.34?x
1 2-2.58?x
2 2-1.51?x
3 2-6.57?x
4 2
Regression analysis and the results of analysis of variance are in Table 1-3, and significant response surface analysis is shown in Fig. 6-Figure 11 mutually alternately.
Table 1-3 returns and the results of analysis of variance
| Source of variation | Quadratic sum | The free degree | All square | F value | P value | ? |
| Model | 2299.26 | 14 | 164.23 | 30.83 | <0.0001 | Extremely remarkable |
| x 1 | 5.56 | 1 | 5.56 | 1.04 | 0.3232 | ? |
| x 2 | 83.51 | 1 | 83.51 | 15.68 | 0.0013 | ? |
| x 3 | 6.86 | 1 | 6.86 | 1.29 | 0.2743 | ? |
| x 4 | 138.38 | 1 | 138.38 | 25.97 | 0.0001 | ? |
| x 1x 2 | 151.35 | 1 | 151.35 | 28.41 | <0.0001 | ? |
| x 1x 3 | 25.78 | 1 | 25.78 | 4.84 | 0.0439 | ? |
| x 1x 4 | 157.31 | 1 | 157.31 | 29.53 | <0.0001 | ? |
| x 2x 3 | 32.92 | 1 | 32.92 | 6.18 | 0.0252 | ? |
| x 2x 4 | 0.10 | 1 | 0.10 | 0.020 | 0.8907 | ? |
| x 3x 4 | 161.35 | 1 | 161.35 | 30.29 | <0.0001 | ? |
| x 1 2 | 354.43 | 1 | 354.43 | 66.52 | <0.0001 | ? |
| x 2 2 | 220.01 | 1 | 220.01 | 41.30 | <0.0001 | ? |
| x 3 2 | 85.17 | 1 | 85.17 | 15.99 | 0.0012 | ? |
| x 4 2 | 1274.72 | 1 | 1274.72 | 239.26 | <0.0001 | ? |
| Residual error | 79.92 | 15 | 5.33 | ? | ? | ? |
| Lose and intend error | 66.24 | 10 | 6.62 | 2.42 | 0.1705 | Not remarkable |
| Pure error | 13.68 | 5 | 2.74 | ? | ? | ? |
| Summation | 2379.17 | 29 | ? | ? | ? | ? |
Note: by analysis, the relative coefficient (R of total regression
2) be 96.64%, the coefficient of determination (R
2 adj) be 93.51%
From showing 1-3, the linear relationship between equation dependent variable and independent variable is obvious, and this model returns extremely significantly (p<0.0001), loses plan item not remarkable, and this model R
2=96.64%, R
2 adj=93.51%, illustrate that this model is good with test matching, between independent variable and response, linear relationship is remarkable, can infer for the theory of this reaction.By F check, can obtain factor contribution rate is: x
4>x
2>x
3>x
1, i.e. ultrasonic time >pH > ultrasound intensity > solid-liquid ratio.
Application responds face optimizing analytical method is analyzed regression model, and when ultrasonic time is 23.91min, pH is 9.72, and ultrasonic power is 378.47W, and material percentage is 0.094, and response surface has optimal value 56.6299 ± 0.42%.。
2.2.4 demonstration test and contrast test
The optimum condition of trying to achieve in response surface analysis method at response surface analysis: when ultrasonic time is 23.91min, pH is 9.72, ultrasonic power is 378.47W, material percentage is under 0.094 condition, carry out 3 parallel tests, the mean value that obtains 3 parallel test black soya bean protein extracting ratios is 56.26%, and the predicted value of minute black soya bean albumen is 56.6299 ± 0.42%, illustrates that the experiment value of response and regression equation predicted value are identical good.
2.4 test brief summaries
Utilizing Response surface methodology from black soya bean, to extract black soya bean albumen ultrasonic processing technique parameter is optimized.Having set up corresponding Mathematical Modeling is that later pilot scale and suitability for industrialized production provided fundamental basis, and has obtained optimum ultrasonic processing technique parameter: ultrasonic time is 23.91min, and pH is 9.72, and ultrasonic power is 378.47W, and material percentage is 0.094.Through checking with contrast test known under optimum ultrasonic processing technique Parameter Conditions black soya bean protein extracting ratio can reach 56.26% left and right.
The Optimum Experiment of test example 2 impulse electric field treatment process parameters
1.1 materials and equipment
1.1.1 raw material and main agents
Black soya bean, it is pure that n-hexane, NaOH, hydrochloric acid etc. are commercially available analysis.
1.1.2 instrument and equipment
| PHS-25 type acidometer | Shanghai great achievement instrument plant |
| Beater disintegrating machine | China Tianjin Stettlen Instrument Ltd. |
| Electric precise mixer | Jintan City, Jiangsu Province high honour instrument manufacturing Co., Ltd |
| Low speed centrifuge | Beijing Medical Centrifugal Machine Factory |
| Supercentrifuge | Germany Eppendorf company |
| Pulse electric field device | Northeast Agricultural University's self-control |
| Electric precise mixer | Jintan City, Jiangsu Province high honour instrument manufacturing Co., Ltd |
| Freeze dryer | U.S. SIM company |
1.3 experimental technique
1.3.1 black soya bean protein preparation method
Raw material black soya bean was pulverized 60 mesh sieves after peeling, fragmentation, then under 40 ℃ of conditions, adopted n-hexane extraction to prepare defatted soybean meal.The mass ratio of defatted soybean meal being pressed to 1:15 mixes with water, with 2mol/L NaOH adjust pH to 8.0, after stirring l.5h, by its suspension centrifugal 30min of 10000 * g under 4 ℃ of conditions, gets 2mol/L HCl adjust pH to 4.5 for supernatant.After standing under 4 ℃ of conditions the centrifugal 30min of 6000 * g, get albumen precipitation washing 2 times, finally get that precipitation is scattered in water and with 2mol/L NaOH adjust pH to 7.0.The centrifugal 30min of 10000 * g under 4 ℃ of conditions, removes a small amount of insoluble matter again, will after its protein solution freeze drying, pulverize and obtain Powdered black soya bean.
1.3.2 technological process (see figure 1)
1.3.3 computing formula
2 results and discussion
2.1 impacts of impulse electric field treatment process list factor condition on black soya bean protein extracting ratio
2.1.1 the impact of pulse width on black soya bean protein extracting ratio
Mixed liquor after ultrasonic processing optimal parameter is processed carries out impulse electric field processing, at sample flow rate, be 60 ml/min, the burst length is 360 μ s, and impulse electric field strength is under 30 kV conditions, investigate the impact of pulse width on black soya bean protein extracting ratio, the results are shown in Figure 12.As shown in Figure 12, along with the increase of pulse width, black soya bean recovery rate significantly raises; But when pulse width reaches 4 μ s, percent of total oil is along with the increase increase of pulse width is not remarkable, and therefore, in response surface experimental design below, pulse width level is selected 3-5 μ s.
2.1.2 the impact of sample flow rate on black soya bean protein extracting ratio
Mixed liquor after ultrasonic processing optimal parameter is processed carries out impulse electric field processing, and pulse width is 4 μ s, and the burst length is 360 μ s, and impulse electric field strength is under 30 kV conditions, investigates the impact of sample flow rate on black soya bean protein extracting ratio, the results are shown in Figure 13.As shown in Figure 13, along with the increase of sample flow rate, black soya bean recovery rate significantly raises; Protein extracting ratio raises along with the increase of sample flow rate, but when sample flow rate reaches 60 ml/min, extraction rate reached is to the highest, and therefore, in response surface experimental design below, sample flow rate level is selected 50-70 ml/min.
2.1.3 the impact of burst length on black soya bean protein extracting ratio
Mixed liquor after ultrasonic processing optimal parameter is processed carries out impulse electric field processing, and pulse width is 4 μ s, and sample flow rate is 360 μ s, and impulse electric field strength is under 30 kV conditions, investigates the impact of burst length on black soya bean protein extracting ratio, the results are shown in Figure 14.As shown in Figure 14, along with the increase in burst length, black soya bean recovery rate significantly raises; But reach after 360 μ s when the burst length, recovery rate raises not obvious, therefore, in response surface experimental design below, burst length level is selected 300-420 μ s.
2.1.4 the impact of impulse electric field strength on black soya bean protein extracting ratio
Mixed liquor after ultrasonic processing optimal parameter is processed carries out impulse electric field processing, and pulse width is 4 μ s, and sample flow rate is 360 μ s, and the burst length is under 360 μ s conditions, investigates the impact of impulse electric field strength on black soya bean protein extracting ratio, the results are shown in Figure 15.As shown in Figure 15, along with the increase of impulse electric field strength, black soya bean recovery rate significantly raises; But when impulse electric field strength reaches after 30 kV, recovery rate raises not obvious, therefore, in response surface experimental design below, impulse electric field strength level is selected 20-40 kV.
2.2 impulse electric fields are processed the response surface Optimum Experiment that extracts black soya bean albumen
2.2.1 experimental factor level code table is in Table 2-1.
On the basis of single factor research, choose pulse width, sample flow rate, burst length, impulse electric field strength, 4 factors are independent variable, take black soya bean protein extracting ratio as response, according to center combination design principle, the experiment of design response surface analysis, its factor level coding schedule is in Table 2-1.
Table 2-1 factor level coding schedule
2.2.2 response surface EE and experimental result
This experimental applications response surface optimization carries out process optimization.With x
1, x
2, x
3, x
4for independent variable, with immobilized enzyme Y, response surface experimental program and the results are shown in Table 2-2.Experiment 1-24 is factorial experiment, and 25-31 is 6 center tests, in order to estimating experiment error.
Table 2-2 response surface experimental program and experimental result
2.2.3 response surface interpretation
By statistical analysis software SAS9.1, carry out data analysis, set up Quadratic response surface regression model as follows:
Y=70.00129+0.593515x1-0.136144x2+0.350832x3-0.656954x4-0.476604x12+0.64195x1x2-1.415312x1x3+0.951899x1x4-1.06025x22+1.840963x2x3-0.084586x2x4-?0.036151?x32+?0.679712x3x4-?1.317635x42
Regression analysis and the results of analysis of variance are in Table 2-3, and response surface optimizing is in Table 2-4, and significant response surface analysis is shown in Figure 16-Figure 21 mutually alternately.
Table 2-3 returns and the results of analysis of variance
| Variable | The free degree | Quadratic sum | All square | F value | Pr>F |
| x 1 | 1 | 8.45423 | 8.45423 | 22.71435 | 0.0002 |
| x 2 | 1 | 0.444843 | 0.444843 | 1.19518 | 0.2905 |
| x 3 | 1 | 2.953997 | 2.953997 | 7.936633 | 0.0124 |
| x 4 | 1 | 10.35814 | 10.35814 | 27.82966 | <.0001 |
| x 1 2 | 1 | 6.49556 | 6.495566 | 17.45192 | 0.0007 |
| x 1x 2 | 1 | 6.593605 | 6.593605 | 17.71533 | 0.0007 |
| x 1x 3 | 1 | 32.04972 | 32.04972 | 86.10938 | <.0001 |
| x 1x 4 | 1 | 14.49779 | 14.49779 | 38.95185 | <.0001 |
| x 2 2 | 1 | 32.14535 | 32.14535 | 86.36633 | <.0001 |
| x 2x 3 | 1 | 54.2263 | 54.2263 | 145.6922 | <.0001 |
| x 2x 4 | 1 | 0.114478 | 0.114478 | 0.307572 | 0.5868 |
| x 3 2 | 1 | 0.037371 | 0.037371 | 0.100406 | 0.7554 |
| x 3x 4 | 1 | 7.392138 | 7.392138 | 19.86078 | 0.0004 |
| x 4 2 | 1 | 49.64682 | 49.64682 | 133.3883 | <.0001 |
| Return | 14 | 214.3619 | 15.31156 | 41.13825 | <.0001 |
| Residue | 6 | 1.834564 | 0.305761 | ? | ? |
| Lose and intend | 10 | 4.1206 | 0.41206 | 1.347655 | 0.3713 |
| |
35 | 2543921 | ? | ? | ? |
Note: by analysis, the relative coefficient (R of total regression
2) be 97.30%, the coefficient of determination (R
2adj) be 94.93%
As shown in Table 3, linear relationship between equation dependent variable and independent variable is obvious, this model returns significantly (p<0.0001), lose plan item not remarkable, and this model R2=97.30%, R2Adj=94.93%, illustrates that this model and experimental fit are good, between independent variable and response, linear relationship is remarkable, can infer for the theory of this reaction.By F check, can obtain factor contribution rate is: x4>x1>x3>x2, i.e. impulse electric field strength > pulse width > burst length > sample flow rate.
Application responds face optimizing analytical method is analyzed regression model, find optimal response the results are shown in Table 2-4 by show 2-4 known when pulse width be 4.3 μ s, sample flow rate is 61 ml/min, the burst length is 369 μ s, when impulse electric field strength is 30kV, response surface has optimal value 70.18 ± 0.23
Table 2-4 response surface optimizing result
| Factor | Level value | Actual value |
| Pulse width x1(μ s) | 0.52020 | 4.260 |
| Sample flow rate x2(ml/min) | 0.22621 | 61.131 |
| Burst length x3(μ s) | 0.15176 | 369.105 |
| Impulse electric field strength x4 (kV) | -0.02951 | 29.852 |
2.2.4 confirmatory experiment and contrast test
Under the optimum condition of trying to achieve in response surface analysis method, when pulse width is 4.3 μ s, sample flow rate is 61 ml/min, burst length is 369 μ s, when impulse electric field strength is 30kV, carry out 3 parallel laboratory tests, black soya bean protein extracting ratio is 70.04%, illustrates that the experiment value of response and regression equation predicted value are identical good.
2.2.4 test brief summary
Mixed liquor after ultrasonic processing optimal parameter is processed carries out impulse electric field processing, utilizes Response surface methodology from black soya bean, to extract black soya bean albumen impulse electric field treatment process parameter and is optimized.Having set up corresponding Mathematical Modeling is that later pilot scale and suitability for industrialized production provided fundamental basis, and obtained optimum impulse electric field treatment process parameter: pulse width is 4.3 μ s, sample flow rate is 61 ml/min, and the burst length is 369 μ s, and impulse electric field strength is 30kV.Through checking with contrast test known under optimum impulse electric field treatment process Parameter Conditions black soya bean protein extracting ratio can reach 70.04% left and right.
Test example 3 is ultrasonic-and impulse electric field processes the impact on black soya bean protein solubility
1 material and equipment
1.1.1 raw material and main agents
Black soya bean, it is pure that n-hexane, NaOH, hydrochloric acid etc. are commercially available analysis, Lowry method protein content determination kit: Li Da bio tech ltd, Shanghai.
1.1.2 instrument and equipment
| PHS-25 type acidometer | Shanghai great achievement instrument plant |
| Beater disintegrating machine | China Tianjin Stettlen Instrument Ltd. |
| Electric precise mixer | Jintan City, Jiangsu Province high honour instrument manufacturing Co., Ltd |
| Low speed centrifuge | Beijing Medical Centrifugal Machine Factory |
| Supercentrifuge | Germany Eppendorf company |
| Pulse electric field device | Northeast Agricultural University's self-control |
| Ultrasonic cell disrupte machine | NingBo XinZhi Biology Science Co., Ltd |
| Electric precise mixer | Jintan City, Jiangsu Province high honour instrument manufacturing Co., Ltd |
| Freeze dryer | U.S. SIM company |
1.3 experimental technique
1.3.1 black soya bean protein preparation method
Raw material black soya bean was pulverized 60 mesh sieves after peeling, fragmentation, then under 40 ℃ of conditions, adopted n-hexane extraction to prepare defatted soybean meal.The mass ratio of defatted soybean meal being pressed to 1:15 mixes with water, with 2mol/L NaOH adjust pH to 8.0, after stirring l.5h, by its suspension centrifugal 30min of 10000 * g under 4 ℃ of conditions, gets 2mol/L HCl adjust pH to 4.5 for supernatant.After standing under 4 ℃ of conditions the centrifugal 30min of 6000 * g, get albumen precipitation washing 2 times, finally get that precipitation is scattered in water and with 2mol/L NaOH adjust pH to 7.0.The centrifugal 30min of 10000 * g under 4 ℃ of conditions, removes a small amount of insoluble matter again, will after its protein solution freeze drying, pulverize and obtain Powdered black soya bean.
1.3.2 technological process (see figure 1)
1.3.3 dissolubility is measured
Take in the deionized water that 100mg protein sample is scattered in 10mL magnetic agitation 30min, 20 ℃, the centrifugal 20min of 12000 * g.Supernatant dilutes through appropriateness, adopts Lowry method to measure protein content, take bovine serum albumin(BSA) as reference material drawing standard curve, sees Figure 22.The solubility table of protein is shown the percentage that supernatant albumen quality concentration accounts for total protein quality concentration
2 results and discussion
2.1 ultrasonic-impulse electric field is processed the impact on black soya bean protein solubility
By previous experiments, to ultrasonic processing and impulse electric field, process optimal parameter assisted extraction black soya bean albumen, measure respectively the heavy albumen of the molten acid of alkali, ultrasonic assisted extraction albumen, impulse electric field assisted extraction albumen, the dissolubility of ultrasonic pulse electric field synergistic assisted extraction albumen, result is as Figure 23, as seen from the figure, ultrasonic processing and impulse electric field are processed and all can be improved to a certain extent the dissolubility of albumen, associated treatment effect is better significantly, protein solubility can reach 62.29%, and the dissolubility of extracting black soya bean albumen than the heavy method of the molten acid of traditional alkali has increased by 16.95%.
Claims (4)
1. the extracting method of a highly dissoluble black soya bean albumen, it is characterized in that the method comprises the following steps: (1) is crossed 60 mesh sieves after black soya bean cleaning is pulverized and obtained black bean powder, black bean powder is mixed in 1:3 ratio with n-hexane, and at room temperature stir 2h and carry out degreasing, after degreasing, centrifugation obtains degreasing black bean powder; (2) degreasing black bean powder and water are mixed to get to mixed liquor, and the mass fraction that regulates the pH of mixed liquor, described black bean powder to account for water is 0.07-0.11, the pH of mixed liquor is 7-11; (3) mixed liquor is carried out to ultrasonic processing, described ultrasonic power is 100-500W, and ultrasonic time is 10-30min; (4) mixed liquor after ultrasonic processing is carried out to impulse electric field processing, described pulse width is 2-6 μ s, and sample flow rate is 40-80ml/min, and the burst length is 60-500 μ s, and impulse electric field strength is 10-40kV; (5) the mixed liquor centrifugal 40min under 10000r/min after impulse electric field processing is obtained to supernatant, supernatant pH is adjusted to 4.5, centrifugation is precipitated, and will precipitate washing and centrifugation repeatedly, obtains the black soya bean albumen of highly dissoluble.
2. the extracting method of a kind of highly dissoluble black soya bean albumen according to claim 1, is characterized in that the mass fraction that described preferred black bean powder accounts for water is 0.094, and preferably the pH of mixed liquor is 9.72.
3. the extracting method of a kind of highly dissoluble black soya bean albumen according to claim 1, is characterized in that described ultrasonic processing preferred parameter is: ultrasonic power 378.47W, ultrasonic time 23.91min.
4. the extracting method of a kind of highly dissoluble black soya bean albumen according to claim 1, is characterized in that described impulse electric field processing preferred parameter is: pulse width 4.3 μ s, sample flow rate 61ml/min, burst lengths 369 μ s, impulse electric field strength 30kV.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107019226A (en) * | 2017-03-10 | 2017-08-08 | 天津和治友德制药有限公司 | A kind of polypeptide powder and its preparation and application |
| CN110037290A (en) * | 2019-05-24 | 2019-07-23 | 福建拓天生物科技有限公司 | A kind of preparation method of coprinus comatus instant powder |
| CN110734469A (en) * | 2018-07-20 | 2020-01-31 | 柴志伟 | Protein alkali-free extraction tower |
| CN111995656A (en) * | 2020-09-11 | 2020-11-27 | 宁波工程学院 | Extraction method of vegetable protein |
| CN112690309A (en) * | 2020-12-25 | 2021-04-23 | 广州市锦华食品有限公司 | Cantonese moon cake containing plant protein meat and making method thereof |
| CN113354749A (en) * | 2021-05-14 | 2021-09-07 | 华南理工大学 | Method for extracting water-soluble soybean polysaccharide from bean dregs by using high-voltage pulse electric field in cooperation with ultrasonic |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101422206A (en) * | 2008-12-05 | 2009-05-06 | 江南大学 | Method for increasing extraction rate of pea separation protein |
| CN101773192A (en) * | 2010-01-14 | 2010-07-14 | 天津大学 | Preparation method of degreased peach kernel protein powder |
| CN102940125A (en) * | 2012-12-07 | 2013-02-27 | 东北农业大学 | Method for preparing high foamability soy protein |
-
2013
- 2013-11-06 CN CN201310541550.XA patent/CN103564145A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101422206A (en) * | 2008-12-05 | 2009-05-06 | 江南大学 | Method for increasing extraction rate of pea separation protein |
| CN101773192A (en) * | 2010-01-14 | 2010-07-14 | 天津大学 | Preparation method of degreased peach kernel protein powder |
| CN102940125A (en) * | 2012-12-07 | 2013-02-27 | 东北农业大学 | Method for preparing high foamability soy protein |
Non-Patent Citations (4)
| Title |
|---|
| 李宝山等: "超声波对大豆分离蛋白提取率及性质的影响", 《饲料研究》 * |
| 李迎秋等: "高压脉冲电场对大豆分离蛋白功能性质的影响", 《农业工程学报》 * |
| 杨敏等: "大豆浓缩蛋白物理改性研究进展", 《粮油食品科技》 * |
| 郭兴凤等: "超声辅助提取黑豆蛋白质的研究", 《河南工业大学学报(自然科学版)》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107019226A (en) * | 2017-03-10 | 2017-08-08 | 天津和治友德制药有限公司 | A kind of polypeptide powder and its preparation and application |
| CN110734469A (en) * | 2018-07-20 | 2020-01-31 | 柴志伟 | Protein alkali-free extraction tower |
| CN110037290A (en) * | 2019-05-24 | 2019-07-23 | 福建拓天生物科技有限公司 | A kind of preparation method of coprinus comatus instant powder |
| CN111995656A (en) * | 2020-09-11 | 2020-11-27 | 宁波工程学院 | Extraction method of vegetable protein |
| CN111995656B (en) * | 2020-09-11 | 2022-09-06 | 宁波工程学院 | Extraction method of vegetable protein |
| CN112690309A (en) * | 2020-12-25 | 2021-04-23 | 广州市锦华食品有限公司 | Cantonese moon cake containing plant protein meat and making method thereof |
| CN113354749A (en) * | 2021-05-14 | 2021-09-07 | 华南理工大学 | Method for extracting water-soluble soybean polysaccharide from bean dregs by using high-voltage pulse electric field in cooperation with ultrasonic |
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