CN102079775A - Method for extracting isolated protein of peanuts - Google Patents
Method for extracting isolated protein of peanuts Download PDFInfo
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- CN102079775A CN102079775A CN 201010559859 CN201010559859A CN102079775A CN 102079775 A CN102079775 A CN 102079775A CN 201010559859 CN201010559859 CN 201010559859 CN 201010559859 A CN201010559859 A CN 201010559859A CN 102079775 A CN102079775 A CN 102079775A
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Abstract
The invention discloses a method for extracting isolated protein of peanuts. The method comprises the following steps of: dissolving peanut protein powder into an alkaline solution, oscillating in a water bath at constant temperature, and centrifuging to obtain supernate 1; extracting precipitate for the second time by the same method, and centrifuging to obtain supernate 2; mixing the supernate 1 and 2, precipitating, and centrifuging to obtain precipitate which is washed with alcohol for twice; and washing with water until neutrality, freezing and drying to obtain the isolated protein of the peanuts. The isolated protein extracted by the method has high content of protein, and the economical benefit and social benefit are remarkable.
Description
Technical field
The present invention relates to peanut food deep process technology field, especially a kind of peanut protein isolate extracting method.
Background technology
China is peanut big producing country, and the peanut annual production occupies first place in the world.Peanut is called as " peanut " in ancient times, often edible can nourish blood enriches blood, tonifying spleen moistening lung, Glycerin, but the disease of preventing hypertension, arteriosclerosis and aspect such as cardiovascular.Protein content in the peanut reaches 20%～30%, is only second to soybean, and peanut also is a kind of important oilseed protein resource.In plant protein resource, Semen arachidis hypogaeae protein occupies the 3rd, accounts for 11% of Tot Prot, is comparatively ideal food protein resource.Semen arachidis hypogaeae protein is the higher vegetable-protein of a kind of nutritive value, it contains 8 kinds of indispensable amino acids of needed by human body, and L-glutamic acid, aspartic acid content are higher, close with animal proteinum, and do not contain cholesterol, digestibility height, its digestibility coefficient reach more than 90% and use wider soybean protein and compare, Semen arachidis hypogaeae protein does not contain the flatulence factor, and antinutritional factor content is less.At present in the world the Semen arachidis hypogaeae protein product according to Protein content, mainly be divided into peanut protein powder (purity＜65%), protein concentrate (purity 65%～70%), protein isolate (purity 85%～96%), wherein peanut protein powder is divided into full-cream, half degreasing and defatted peanut protein powder again.The peanut protein isolate preparation method mainly comprises alkali extraction and acid precipitation, ultrafiltrationmembrane process, ultrasonic extraction and aqueous enzymatic method.Research mainly concentrates on: aspects such as peanut protein powder, peanut protein beverage, condensed milk of peanut protein, Semen arachidis hypogaeae protein film, peanut polypeptide and Semen arachidis hypogaeae protein modification to the Semen arachidis hypogaeae protein Products Development in China.The isolating Semen arachidis hypogaeae protein purity of prior art is low, and the albumen yield is low, and the production time is long.Therefore, need seek a kind of extraction yield height and the high separation method of protein isolate purity.
Summary of the invention
The object of the present invention is to provide a kind of peanut protein isolate extracting method, to improve the yield of peanut concentrated protein.
For achieving the above object, the present invention realizes by the following technical solutions:
Peanut protein powder is dissolved in the NaOH solution of pH value 10, solid-liquid ratio dissolving in 1: 10～1: 12 fully, 40～50 ℃ of water-baths vibrations, lixiviate 90～150min, centrifugal, get supernatant liquor 1; To precipitate secondary lixiviate in the same way, centrifugal, get supernatant liquor 2; Supernatant liquor 1 and 2 is merged, the salt Acid precipitation, centrifugal, taking precipitate alcohol is washed 2 times, is washed to neutrality again, and lyophilize gets peanut protein isolate.
Preferably, separation method of the present invention is:
Peanut protein powder is dissolved in the NaOH solution of pH value 10, solid-liquid ratio (weightmeasurement ratio) dissolving in 1: 12 fully, 50 ℃ of water-baths vibrations, lixiviate 131min, centrifugal, get supernatant liquor 1; To precipitate secondary lixiviate in the same way, centrifugal, get supernatant liquor 2; Supernatant liquor 1 and 2 is merged, the salt Acid precipitation, centrifugal, taking precipitate alcohol is washed 2 times, is washed to neutrality again, and lyophilize gets peanut protein isolate.
Utilize isolating Semen arachidis hypogaeae protein extraction rate of protein of present method and content height, detect through Kjeldahl determination, the purity of peanut protein isolate is more than 90%, more than the extraction rate reached to 65%, and economic benefit and obvious social benefit.
Description of drawings
Fig. 1 extraction temperature is to the influence of peanut protein isolate preparation
Fig. 2 alkali lye pH value is to the influence of peanut protein isolate preparation
Fig. 3 extraction time is to the influence of peanut protein isolate preparation
Fig. 4 solid-liquid ratio is to the influence of peanut protein isolate preparation
The response surface figure of Fig. 5 dual factors
A. extraction time and temperature are to the influence of purity of protein
B. extraction time and alkali lye pH are to the influence of purity of protein
C. the influence of extraction time and liquid material comparison purity of protein
D. extraction temperature and alkali lye pH are to the influence of purity of protein
E. the influence of extraction temperature and liquid material comparison purity of protein
F. the influence of alkali lye pH and liquid material comparison purity of protein
Preparation technology's flow process of Fig. 6 peanut protein isolate
Experimental example
1, materials and methods
1.1 material and reagent
One-level defatted peanut protein powder: Shandong Tianshen Bioprotein Co., Ltd.
It is pure that 95% ethanol, distilled water, hydrochloric acid, NaOH, boric acid etc. are analysis. Methyl red, bromocresol green indicator.
1.2 instrument and equipment
Freeze drier Beijing rich doctor health laboratory apparatus Co., Ltd; The difunctional water-bath constant temperature oscillator of SHA-B Jintan City Jie Ruier Electrical Appliances Co., Ltd; FE20 laboratory pH meter plum Teller-Tuo benefit Instr Ltd.; The DZ-2 automatic potentiometric titrimeter; Full-automatic Kjeldahl determination device
1.3 experimental technique
1.3.1 experiment of single factor design
Change successively that peanut protein powder alkali is put forward the temperature of Acid precipitation, alkali lye pH value, alkali carries the time and alkali is put forward solid-to-liquid ratio, carry out research and analysis with Semen arachidis hypogaeae protein purity and albumen yield as evaluation index, and the optimum parameter of definite four factors, three levels is carried out response surface analysis.
1.3.2 the mensuration of purity of protein and albumen yield
Purity of protein: Kjeldahl determination
Total protein quality * 100% in total protein quality/peanut protein powder in protein extracting ratio (%)=gained peanut protein isolate
1.3.3 feed composition Determination on content
The mensuration of protein content: GB/T5009,5-2003; Determination of fat: GB/T5009,6-2003
The mensuration of moisture content: GB/T5009,3-2003; The mensuration of ash oontent: GB/T5009,4-2003
2 results and analysis
2.1 the main component of peanut protein powder
The main component of table 1 peanut protein powder
2.2 experiment of single factor result and analysis
2.2.1 extraction temperature is to the influence of peanut protein isolate preparation
With the pH value of the hierarchy of control in the process of alkali lye lixiviate peanut protein powder be 9, extraction time is that 90min, solid-to-liquid ratio are 1: 10, different temperature is set, but temperature is unsuitable too high, in order to avoid make protein denaturation, the research extraction temperature is to the influence (Fig. 1) of peanut protein isolate preparation.As we know from the figure, along with the rising of temperature, purity of protein improves gradually, and purity of protein is tending towards constant more than 40 ℃; Protein extracting ratio raises with temperature, raises earlier and reduces, and protein extracting ratio is the highest in the time of 40 ℃.Take all factors into consideration two evaluation indexes and determine that temperature is 40,45,50 ℃ and continues to do response surface analysis to determine best extraction temperature.
2.2.2 alkali lye pH value is to the influence of peanut protein isolate preparation
The temperature of the hierarchy of control is that 40 ℃, solid-to-liquid ratio are that 1: 10, extraction time are 90min in the alkali lye lixiviate peanut protein powder process, and the alkali lye of different pH values is set.Record experiment before the experiment with the water-soluble apparent acidity of peanut protein powder, when protein powder is dissolved in the alkali lye of pH value≤10.5, show weakly alkaline or neutrality, can not make protein denaturation.Alkali lye pH value the results are shown in shown in Figure 2 to the influence of peanut protein isolate preparation.The Semen arachidis hypogaeae protein main component is arachin and Ban Huashengqiudanbai, also has about 10% white protein.Whole sphaeroprotein and white protein all dissolve in the dilute alkaline soln, and protein solubility raises with pH and increases, and the protein stripping increases.As we know from the figure, along with alkali lye pH value increases, purity of protein and extraction yield all are the trend of increase, and wherein pH value=10 o'clock protein extracting ratio is the highest, and protein content tends towards stability when pH=9.Take all factors into consideration the optimal ph point of two evaluation indexes, choose pH and be 9,9.5,10 and continue to do response surface analysis to determine best alkali lye pH value.
2.2.3 extraction time is to the influence of peanut protein isolate preparation
The temperature of the hierarchy of control is that 40 ℃, solid-to-liquid ratio are that 1: 10, alkali lye pH value=9.5 are provided with different extraction times in the alkali lye lixiviate peanut protein powder process, and the research extraction time the results are shown in Figure 3 to the influence of peanut protein isolate preparation.As shown in the figure, purity of protein increases along with the increase of time, and increasing degree is slowed down after the 90min, and raising descends again and protein extracting ratio increases earlier in time.This process may be that extraction time prolongs, and proteic stripping increases, behind certain hour, proteic stripping reaches capacity, then solubility rate tends to balance, if further prolong extraction time again, thus may be because factors such as microorganism growth make protein denaturation that extraction yield is reduced.The best extraction time section of comprehensive evaluation two indexes is chosen 90min, 120min, 150min and is continued to do response surface analysis to determine best extraction time.
2.2.4 solid-liquid ratio is to the influence of peanut protein isolate preparation
The temperature of the hierarchy of control is that 40 ℃, extraction time are 90min, alkali lye pH value=9.5 in the alkali lye lixiviate peanut protein powder process, and different solid-to-liquid ratios is set, and the research solid-to-liquid ratio the results are shown in Figure 4 to the influence of peanut protein isolate preparation.As shown in the figure, purity of protein changes not quite with the increase of solid-to-liquid ratio, and protein extracting ratio then increases with solid-to-liquid ratio and improves.Because unit alkali lye can only dissolve a certain amount of albumen, the alkali lye amount that this experiment of single factor is selected for use can not make Semen arachidis hypogaeae protein fully dissolve or Semen arachidis hypogaeae protein dissolving fully just, so under different solid-to-liquid ratio conditions, purity of protein tends to balance.And increase with the alkali lye amount, proteic meltage increases, thereby makes proteic extraction yield increase.As can be seen from the figure, be the sufficient point of protein dissolution about 1: 11, continue to do response surface analysis to determine best solid-liquid ratio so choose solid-to-liquid ratio=1: 10,1: 11,1: 12 in solid-to-liquid ratio.
2.3 the optimization of peanut protein isolate extraction process
According to adopting Box-Behnken center combination experimental design principle, in conjunction with the single factor experiment result, investigate the influence to purity of protein (Y) of extraction time (X1), temperature (X2), alkali lye pH value (X3) and solid-liquid ratio (X4), experimental factor and level see Table 2, test design and the results are shown in Table 3.Test design adopts Design-expert software with analysis.
Table 2 center combination experimental factor level code table
Table 3 test design and result
2.3.1 the foundation of model and test of significance
According to testing data, independent variable(s) coding X1, X2, X3 and X4 are carried out regression analysis, must the multinomial regression equation of secondary be:
Y＝93.85+1.02X1+0.67X2+0.97X3-0.47X4+0.76X1X2-0.85X1X3+0.42X1X4-0.32X2X3+0.55X2X4+0.91X3X4-1.76X12+0.084X22-0.19X32+0.010X42
Carry out the significance that regression model and each parameter are verified in variance analysis by Design-Expert software, the results are shown in Table 4
The ANOVA analytical results of table 4 model
Annotate: the P value has remarkably influenced less than 0.05 explanation model or investigation factor; The P value is extremely remarkable less than 0.01 explanation influence.
By variance analysis as can be seen, model P value is much smaller than 0.05, and the representation model equation is extremely remarkable, and it is not remarkable that model loses the plan item, and Model Selection is fit to.The degree of confidence of variation coefficient reflection model, the variation coefficient is low more, and the degree of confidence of model is good more; The variation coefficient of this test is 1.07%, illustrates that model equation can well react real trial value.Therefore can with this model equation analyze with predict different extracting conditions under, the changing conditions of Semen arachidis hypogaeae protein purity.P value by table 4 can know, X1, X2 in the equation, X3, X12 are remarkable to the influence of Y, show that experimental factor is not linear to the influence of response value.By the direct size of a coefficient absolute value once in the equation relatively, primary and secondary that can the factor of judgment influence.Size to the proteic purity influence of extraction separation is extraction time, alkali lye pH value, temperature, solid-liquid ratio successively, and promptly extraction time is the most remarkable to the extraction influence of protein isolate.
The figure of RSM is the circle of equal altitudes of a three-dimensional space on two dimensional surface that the specific corresponding factor X1 of response surface Y, X2, X3, X4 value constitutes, of the influence of each factor can be reflected intuitively, the interaction between them can be analyzed from the gained response surface analysis figure response value.The relation of response value and influence factor as can be seen from response surface analysis Fig. 5.
A-f has reflected the influence of each factor to response value intuitively among Fig. 5, exists the condition of extreme value should be at circle centre position as can be seen by isogram.Relatively 6 picture groups as can be known: extraction time is the most remarkable to the influence of purity of protein, and the performance curve is steeper; And alkali lye pH, extraction temperature, solid-liquid ratio take second place, and it is comparatively level and smooth to show as curve, and with the increase or the minimizing of its numerical value, response value changes less.
2.3.2 the optimization of extraction conditions and checking
Further trial model is carried out the typicalness analysis, to obtain optimum extraction conditions with Design-Expert software.By analysis, at X1=130.77min, X2=50 ℃, X3=10, X4=12, promptly the theoretical maximum that obtains at 1: 12 of extraction time 130.77min, 50 ℃ of temperature, alkali lye pH value 10, solid-liquid ratio is 96.32%
In order to consider the feasibility of proof test, the optimum extraction condition that employing obtains carries out the extraction test of peanut protein isolate, consider operation and the convenience of producing simultaneously, extraction conditions is modified to extraction time X1=131min, other conditions are constant, the purity of protein average out to 95.05% that 3 parallel tests obtain, differ 1.27% with theoretical value, therefore the response surface method is feasible to the optimization of peanut protein isolate extraction conditions, and the peanut protein isolate extraction conditions that obtains has actual application value.
3 conclusions
This experiment is the response surface analysis method of carrying out four factors, three levels on single factor basis, the processing condition of extracting peanut protein isolate have been optimized, the response surface experimental analysis goes out, be 90～150min in extraction time, extract that temperature is that 40～50 ℃, alkali lye pH value are 9～10, solid-liquid ratio is that each factor is extraction time, alkali lye pH value, temperature, solid-liquid ratio to the size that the proteic purity of extraction separation influences successively in 1: 10～1: 12 the scope; Extracted peanut protein isolate optimum extraction process parameter and be extraction time 131min, 50 ℃ of temperature, alkali lye pH value 10, solid-liquid ratio 1: 12.
Embodiment
Below in conjunction with embodiment the present invention is described in detail.
Embodiment 1
Peanut protein powder is dissolved in the NaOH solution of pH value 10, solid-liquid ratio dissolving in 1: 10 fully, 40 ℃ of water-baths vibrations, lixiviate 90min, centrifugal, get supernatant liquor 1; To precipitate secondary lixiviate in the same way, centrifugal, get supernatant liquor 2; Supernatant liquor 1 and 2 is merged, the salt Acid precipitation, centrifugal, taking precipitate alcohol is washed 2 times, is washed to neutrality again, and lyophilize gets peanut protein isolate.
Detect through Kjeldahl determination, the purity of peanut protein isolate is 90.02%, extraction rate reached to 66.05%.
Embodiment 2
Peanut protein powder is dissolved in the NaOH solution of pH value 10, solid-liquid ratio dissolving in 1: 11 fully, 45 ℃ of water-baths vibrations, lixiviate 150min, centrifugal, get supernatant liquor 1; To precipitate secondary lixiviate in the same way, centrifugal, get supernatant liquor 2; Supernatant liquor 1 and 2 is merged, the salt Acid precipitation, centrifugal, taking precipitate alcohol is washed 2 times, is washed to neutrality again, and lyophilize gets peanut protein isolate.
Detect through Kjeldahl determination, the purity of peanut protein isolate is 92.15%, extraction rate reached to 67.51%.
Embodiment 3
Peanut protein powder is dissolved in the NaOH solution of pH value 10, solid-liquid ratio dissolving in 1: 12 fully, 50 ℃ of water-baths vibrations, lixiviate 131min, centrifugal, get supernatant liquor 1; To precipitate secondary lixiviate in the same way, centrifugal, get supernatant liquor 2; Supernatant liquor 1 and 2 is merged, the salt Acid precipitation, centrifugal, taking precipitate alcohol is washed 2 times, is washed to neutrality again, and lyophilize gets peanut protein isolate.
Detect through Kjeldahl determination, the purity of peanut protein isolate is 95.05%, extraction rate reached to 71%
Peanut protein powder is dissolved in the NaOH solution of pH value 10, solid-liquid ratio dissolving in 1: 12 fully, 48 ℃ of water-baths vibrations, lixiviate 140min, centrifugal, get supernatant liquor 1; To precipitate secondary lixiviate in the same way, centrifugal, get supernatant liquor 2; Supernatant liquor 1 and 2 is merged, the salt Acid precipitation, centrifugal, taking precipitate alcohol is washed 2 times, is washed to neutrality again, and lyophilize gets peanut protein isolate.
Detect through Kjeldahl determination, the purity of peanut protein isolate is 94.76%, extraction rate reached to 70.80%.
Claims (3)
1. the preparation method of a peanut protein isolate may further comprise the steps:
Peanut protein powder is dissolved in the NaOH solution of pH value 10, solid-liquid ratio dissolving in 1: 10～1: 12 fully, 40～50 ℃ of water-baths vibrations, lixiviate 90～150min, centrifugal, get supernatant liquor 1; To precipitate secondary lixiviate in the same way, centrifugal, get supernatant liquor 2; Supernatant liquor 1 and 2 is merged, the salt Acid precipitation, centrifugal, taking precipitate alcohol is washed 2 times, is washed to neutrality again, and lyophilize gets peanut protein isolate.
2. the preparation method of peanut protein isolate according to claim 1 is characterized in that, may further comprise the steps:
Peanut protein powder is dissolved in the NaOH solution of pH value 10, solid-liquid ratio dissolving in 1: 12 fully, 50 ℃ of water-baths vibrations, lixiviate 131min, centrifugal, get supernatant liquor 1; To precipitate secondary lixiviate in the same way, centrifugal, get supernatant liquor 2; Supernatant liquor 1 and 2 is merged, the salt Acid precipitation, centrifugal, taking precipitate alcohol is washed 2 times, is washed to neutrality again, and lyophilize gets peanut protein isolate.
3. the preparation method of peanut protein isolate according to claim 2 is characterized in that, described alcohol is 95% ethanol.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102239954A (en) * | 2011-07-01 | 2011-11-16 | 华南理工大学 | Preparation and decolouring methods of peanut proteins |
| CN102757489A (en) * | 2012-07-24 | 2012-10-31 | 中国农业科学院农产品加工研究所 | Method for synchronously extracting and separating arachin and conarrachin |
| CN103766574A (en) * | 2013-12-31 | 2014-05-07 | 山东省花生研究所 | Process for producing peanut protein isolate by virtue of enzymatic modification |
| CN106046142A (en) * | 2016-06-24 | 2016-10-26 | 江苏省农业科学院 | Method for separating protein from pig livers |
| CN108892704A (en) * | 2018-07-18 | 2018-11-27 | 辽宁大学 | One extracting method and its application for cultivating peanut leaf soluble protein |
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| CN101543256A (en) * | 2009-04-30 | 2009-09-30 | 山东省高唐蓝山集团总公司 | Process technology for automatically producing peanut protein isolate |
| CN101731445A (en) * | 2010-02-06 | 2010-06-16 | 赵广彬 | Method for preparing peanut protein and peanut peptide by using low temperature peanut pulp |
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2010
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| CN101543256A (en) * | 2009-04-30 | 2009-09-30 | 山东省高唐蓝山集团总公司 | Process technology for automatically producing peanut protein isolate |
| CN101731445A (en) * | 2010-02-06 | 2010-06-16 | 赵广彬 | Method for preparing peanut protein and peanut peptide by using low temperature peanut pulp |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102239954A (en) * | 2011-07-01 | 2011-11-16 | 华南理工大学 | Preparation and decolouring methods of peanut proteins |
| CN102239954B (en) * | 2011-07-01 | 2013-03-06 | 华南理工大学 | Preparation and decolouring methods of peanut proteins |
| CN102757489A (en) * | 2012-07-24 | 2012-10-31 | 中国农业科学院农产品加工研究所 | Method for synchronously extracting and separating arachin and conarrachin |
| CN103766574A (en) * | 2013-12-31 | 2014-05-07 | 山东省花生研究所 | Process for producing peanut protein isolate by virtue of enzymatic modification |
| CN106046142A (en) * | 2016-06-24 | 2016-10-26 | 江苏省农业科学院 | Method for separating protein from pig livers |
| CN108892704A (en) * | 2018-07-18 | 2018-11-27 | 辽宁大学 | One extracting method and its application for cultivating peanut leaf soluble protein |
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Application publication date: 20110601 |