Method for extracting component protein of low-gluten rice
阅读说明：本技术 一种低谷蛋白稻米组分蛋白的提取方法 (Method for extracting component protein of low-gluten rice ) 是由 李天� 兰艳 隋晓东 刘泉 涂云彪 孙影影 王锦 龚静 吴超越 于 2019-05-23 设计创作，主要内容包括：本发明公开了一种低谷蛋白稻米组分蛋白的提取方法,包括以下步骤：1)准确称取精米粉,水浴振荡浸提,以6000rpm转速离心10min,收集上清液,为清蛋白溶液；2)将步骤(1)提取过的稻米中加入氯化钠溶液,水浴振荡浸提,以6000rpm转速离心10min,收集上清液,为球蛋白溶液；3)将步骤(2)提取后稻米加入乙醇溶液,水浴振荡浸提,以6000rpm转速离心10min,收集上清液,为醇溶蛋白溶液；4)将步骤(3)提取后稻米加入氢氧化钠溶液,水浴振荡浸提,以6000rpm转速离心10min,收集上清液,为谷蛋白溶液。该方法操作简单,为低谷蛋白功能稻的综合评价提供一定的理论依据。(The invention discloses a method for extracting component protein of low-gluten rice, which comprises the following steps: 1) accurately weighing fine rice flour, extracting in water bath under oscillation, centrifuging at 6000rpm for 10min, and collecting supernatant as albumin solution; 2) adding a sodium chloride solution into the rice extracted in the step (1), carrying out water bath oscillation leaching, centrifuging at 6000rpm for 10min, and collecting supernatant as a globulin solution; 3) adding the rice extracted in the step (2) into an ethanol solution, carrying out water bath oscillation extraction, centrifuging at 6000rpm for 10min, and collecting supernatant as an alcohol soluble protein solution; 4) and (4) adding the rice extracted in the step (3) into a sodium hydroxide solution, carrying out water bath oscillation leaching, centrifuging at the rotating speed of 6000rpm for 10min, and collecting the supernatant as a gluten solution. The method is simple to operate, and provides a certain theoretical basis for the comprehensive evaluation of the low-gluten functional rice.)
1. A method for extracting the component protein of low-gluten rice is characterized by comprising the following steps:
1) accurately weighing fine rice flour, shaking and leaching in a constant-temperature water bath shaker, centrifuging at 6000rpm for 10min, and collecting supernatant as albumin solution;
2) adding a sodium chloride solution into the rice extracted in the step (1), oscillating and leaching by a constant-temperature water bath shaker, centrifuging at 6000rpm for 10min, and collecting supernatant as a globulin solution;
3) adding the rice extracted in the step (2) into an ethanol solution, carrying out oscillating extraction on the rice by a constant-temperature water bath shaker, centrifuging the rice for 10min at the rotating speed of 6000rpm, and collecting supernatant to obtain an alcohol soluble protein solution;
4) and (4) adding the rice extracted in the step (3) into a sodium hydroxide solution, carrying out oscillating extraction on the rice by a constant-temperature water bath shaking table, centrifuging the rice at the rotating speed of 6000rpm for 10min, and collecting supernatant to obtain a gluten solution.
2. The method for extracting the component protein of the low gluten rice as claimed in claim 1, wherein the mass ratio of the feed liquid in the step (1) is 1:22, the extraction temperature is 31.6 ℃, the extraction time is 43min, and the extraction times are 3 times.
3. The method for extracting the component protein of the low gluten rice as claimed in claim 1, wherein the mass ratio of the feed liquid in the step (2) is 1:20, the extraction temperature is 33.1 ℃, the extraction time is 22min, and the extraction times are 3 times.
4. The method for extracting the component protein of the low gluten rice as claimed in claim 1, wherein the mass ratio of the feed liquid in the step (3) is 1:21, the extraction temperature is 34.6 ℃, the extraction time is 40min, and the extraction times are 2 times.
5. The method for extracting the component protein of the low gluten rice as claimed in claim 1, wherein the mass ratio of the feed liquid in the step (4) is 1:16, the extraction temperature is 34.7 ℃, the extraction time is 22min, and the extraction times are 3 times.
Technical Field
The invention belongs to the technical field of proteomics extraction, and particularly relates to a method for extracting component proteins of low-gluten rice.
Background
The rice grains contain 7-10% of protein, the protein content of the general brown rice is about 8%, and the protein content of the polished rice is about 7%. Proteins can be classified into four categories according to their different solubilities: water-soluble albumin (albumin), salt-soluble globulin (globulin), alcohol-soluble prolamin (prolamin) and alkali-soluble gluten (gluten), which account for 5%, 10%, 5% and 80% of the rice protein, respectively. Albumin and globulin are mainly stored in the tissues of the peel, aleurone layer and embryo, while prolamin and gluten are mainly stored in the endosperm. Gluten is stored in the protein body PB-II and is easily digested and absorbed by the stomach of a human body, and alcohol soluble protein is deposited in the protein body PB-I and cannot be digested and absorbed by the stomach, so that the human body mainly absorbs the gluten. Patients with diabetes and kidney diseases cannot eat rice with soluble protein content of more than 4% due to renal dysfunction. The low glutelin rice is a rice variety which can absorb 3.1 to 4 percent of protein (the sum of albumin, globulin and glutelin), and is also called low water-solubility rice. After a patient eats the same amount of the low-gluten rice, the blood sugar and the blood fat of the human body can not be increased due to excessive heat intake, and the diabetes and the nephropathy can be effectively prevented and treated in an auxiliary way.
Researches suggest that the protein components in rice grains have obvious difference among genotypes. In recent years, with the increasing number of patients with kidney diseases and diabetes complicated with kidney function damage, the market demand of functional rice suitable for such special population is increasing. At present, the domestic research on the low-gluten rice has made a certain progress in breeding and cultivation, but the determination of the component protein thereof is continued to use the continuous extraction method of the common rice. The content of each protein component in the low-gluten rice is far from that of the common rice, and the extraction condition of the common rice is not suitable.
Disclosure of Invention
The invention aims to provide a method for extracting component protein of low-gluten rice, which provides a certain theoretical basis for the comprehensive evaluation of low-gluten functional rice.
The invention is realized by the following technical scheme:
a method for extracting the component protein of low-gluten rice comprises the following steps:
1) accurately weighing fine rice flour, shaking and leaching in a constant-temperature water bath shaker, centrifuging at 6000rpm for 10min, and collecting supernatant as albumin solution;
2) adding a sodium chloride solution into the rice extracted in the step (1), oscillating and leaching by a constant-temperature water bath shaker, centrifuging at 6000rpm for 10min, and collecting supernatant as a globulin solution;
3) adding the rice extracted in the step (2) into an ethanol solution, carrying out shaking extraction on the rice by a constant-temperature water bath shaker, centrifuging the rice for 10min at the rotating speed of 6000rpm, and collecting supernatant to obtain an alcohol soluble protein solution;
4) and (4) adding the rice extracted in the step (3) into a sodium hydroxide solution, carrying out oscillating extraction on the rice by a constant-temperature water bath shaking table, centrifuging the rice for 10min at the rotating speed of 6000rpm, and collecting supernatant to obtain a gluten solution.
Further, in the step (1), the mass ratio of the feed liquid is 1:22, the extraction temperature is 31.6 ℃, the extraction time is 43min, and the extraction times are 3 times.
Further, in the step (2), the material liquid mass ratio is 1:20, the extraction temperature is 33.1 ℃, the extraction time is 22min, and the extraction times are 3 times.
Further, in the step (3), the material liquid mass ratio is 1:21, the extraction temperature is 34.6 ℃, the extraction time is 40min, and the extraction times are 2 times.
Further, in the step (4), the mass ratio of the feed liquid is 1:16, the extraction temperature is 34.7 ℃, the extraction time is 22min, and the extraction times are 3 times.
The invention has the beneficial effects that:
the invention provides a method for extracting the component protein of low-gluten rice, which is simple to operate, can respectively extract albumin, globulin, alcohol soluble protein and gluten, is not interfered with each other, and has popularization and use values.
Drawings
FIG. 1 is a graph of the effect of different extractant concentrations on the protein content of each component; a is globulin, B is prolamin, C is glutelin;
FIG. 2 is a graph showing the effect of feed liquid ratio on protein content of each component; a is albumin, B is globulin, C is prolamin, D is glutelin;
FIG. 3 is a graph of the effect of temperature on the protein content of each component; a is albumin, B is globulin, C is prolamin, D is glutelin;
FIG. 4 is a graph showing the effect of extraction time on protein content of each fraction; a is albumin, B is globulin, C is prolamin, D is glutelin;
FIG. 5 is the effect of the number of extractions on the protein content of each fraction; a is albumin, B is globulin, C is prolamin, D is glutelin;
FIG. 6 is a 3D response profile of the effect of 4-factor interaction on albumin content (A-D, C-D, B-C, B-D, A-B, A-C);
FIG. 7 is a 3D response profile of the effect of 4-factor interaction on globulin content (A-D, C-D, B-C, B-D, A-B, A-C);
FIG. 8 is a 3D response profile of the effect of 4-factor interaction on prolamin content (A-D, C-D, B-C, B-D, A-B, A-C);
FIG. 9 is a 3D response profile of the effect of 4-factor interaction on gluten content (A-D, C-D, B-C, B-D, A-B, A-C).
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to specific embodiments of the present invention, and it should be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
1 materials and methods
1.1 materials and reagents
Selecting low-gluten rice (D105) as a material under the same field cultivation condition, collecting the rice, drying in the air, storing for three months, processing into fine rice, crushing, sieving, and drying for testing.
G250, bovine serum albumin standard, absolute ethyl alcohol, sodium chloride (AR), phosphoric Acid (AR) and sodium hydroxide (AR) were purchased from Durett technologies, Inc., Sichuan.
1.2 instruments and devices
A pulverizer (Yunpong YB-750A), a constant temperature water bath shaking table (THZ-82A), a Centrifuge (ThermosCIENTIFIC SOLVALLY BIOFUGE STRATOS Centrifuge), a double-beam ultraviolet spectrophotometer (Germany Analytik Jena Specord 200Plus), an ultra-pure water machine (YouPUULUP series), and a BT-124S type electronic balance (Germany Sartorius company).
1.3 test methods
1.3.1 extractant concentration screening test
Taking the protein content of each component as an index, investigating the concentration of an extracting agent: influence of globulin extractant sodium chloride solution (0.5, 0.6, 0.7, 0.8, 0.9, 1.0mol/L), prolamine extractant ethanol (70%, 75%, 80%, 85%, 90%), gluten extractant sodium hydroxide solution (0.02, 0.04, 0.06, 0.08, 0.09, 0.1mol/L) on protein content of each component, and fixed temperature of 35 ℃, time of 30min, extraction for 3 times.
1.3.2 component protein content determination
The extraction amount of component protein in rice is determined by G250 color development, taking 1mL of solution to be detected, adding 4mLG250 reagent, shaking, standing for 5min, and detecting light absorption value at 595 nm. And drawing a standard curve by taking the concentration of the bovine serum protein standard solution as an x axis and the absorbance as a y axis.
The protein content of each component is calculated as the mass concentration of protein in the sample liquid, the volume of the liquid to be detected, the fixed volume/the weight of the rice flour/1000.
1.3.3 extraction of proteins of Components in Rice
The component protein of the rice is extracted by adopting a continuous extraction method, which is slightly changed. Accurately weighing 0.300g of fine rice flour, adding water, shaking and leaching in a constant temperature water bath shaker (190rpm), centrifuging for 10min at 6000rpm, collecting supernatant, and diluting to constant volume to obtain albumin solution to be detected; adding sodium chloride solution into the sample after albumin extraction, carrying out oscillation leaching by a constant temperature water bath shaker (190rpm), carrying out centrifugation for 10min at 6000 revolutions, collecting supernatant, and fixing the volume to obtain a solution to be detected of globulin; adding an ethanol solution into the sample after the globulin is extracted, similarly performing the steps, collecting supernatant, and performing constant volume to obtain a solution to be detected of the prolamin; and adding a sodium hydroxide solution into the sample extracted with the over-alcohol-soluble protein, collecting the supernatant, and fixing the volume to obtain a gluten solution to be detected. The content of each component protein in the rice is measured and calculated according to the method 1.3.2.
1.3.4 Single factor test
The influence of the ratio of feed to liquid (1:10, 1:15, 1:20, 1:25, 1:30, fixed
1.3.5 response surface optimization test method
On the basis of a single-factor test result, Design-Expert11(RSM) software is adopted to Design test conditions, a Box-Behnken (BB) model is selected, the protein content of each component is used as a response value, the material-liquid ratio (A), the extraction temperature (B), the extraction time (C) and the extraction frequency (D) are used as independent variables, and a 4-
TABLE 1 factor level coding scheme
1.4 data processing
Statistical analysis was performed on experimental data using Microsoft Excel 2010, SPSS 17.0, Design expert11, SAS 9.4. All experiments were repeated three times.
2 results and analysis
2.1 test results of extractant concentration screening
2.1.1 Effect of sodium chloride concentration on globulin content
As shown in FIG. 1A, the globulin content gradually increased with the increase of the sodium chloride concentration, and when the sodium chloride concentration was 0.6mol/L, the content reached 6.29mg/g, and the concentration continued to increase, but the content gradually decreased. This may be a partial precipitation of protein by higher concentrations of salt ions, reducing the amount of globulin extracted. Therefore, the sodium chloride concentration was selected to be 0.6 mol/L.
2.1.2 Effect of ethanol concentration on prolamin content
As shown in FIG. 1B, the prolamin content gradually increased with the increase of ethanol concentration, and when the ethanol concentration was 80%, the prolamin content reached 4.13mg/g, and the concentration continued to increase, but gradually decreased. This may be that higher ethanol concentrations cause structural disruption of the protein, resulting in partial precipitation, thereby reducing prolamin extraction. Therefore, an ethanol concentration of 80% was selected.
2.1.3 Effect of sodium hydroxide concentration on gluten content
As can be seen from FIG. 1C, the gluten content gradually increased with the increase of the NaOH concentration, and reached 14.15mg/g at 0.04mol/L, while the concentration continued to increase and gradually decreased. This is probably because sodium hydroxide reacts somewhat with the protein and at higher concentrations it reduces the solubility of the protein and thus the amount of gluten extracted. Therefore, the sodium hydroxide concentration was selected to be 0.04 mol/L.
2.2 Single factor test results
2.2.1 Effect of feed liquid ratio on protein content of Each component
As can be seen from FIG. 2, with the increase of the liquid-to-material ratio, the protein extraction amount of the four components is increased, and albumin, globulin, prolamine and gluten reach maximum values of 5.59mg/g, 15.27mg/g, 4.19mg/g and 20.26mg/g at the liquid-to-material ratios of 1:20, 1:20, 1:25 and 1:15, respectively. When the extraction liquid-material ratio exceeds the liquid-material ratio of the maximum extraction value, the extraction amount of the four-component protein is gradually reduced. This is probably because the liquid-material ratio is 1:20, 1:20, 1:25, 1:15, four proteins of rice are basically dissolved out, the liquid-material ratio is continuously increased, soluble impurities are increased, affinity of the proteins to a solvent is reduced, and the extraction amount is reduced. Therefore, albumin was selected for response surface analysis: 1:15, 1:20, 1: 25; globulin: 1:15, 1:20, 1: 25; prolamin: 1:20, 1:25, 1: 30; gluten: the ratio of 1:10, 1:15 and 1:20 are proper liquid-material ratios.
2.2.2 Effect of extraction temperature on protein content of Components
As can be seen from FIG. 3, when the time is 25-40 ℃, the protein extraction amount of the four components is increased, and albumin, globulin and alcohol soluble all reach the maximum value of 4.59mg/g (albumin), 10.29mg/g (globulin), 2.49mg/g (alcohol soluble protein) at 35 ℃, and gluten reaches the maximum value of 19.68mg/g at 40 ℃. When the extraction temperature exceeded 35 ℃ (albumin, prolamin, globulin) and 40 ℃ (gluten), the extraction of the four component proteins gradually decreased. This is probably because the certain temperature (35, 40 ℃) in the test accelerates the intermolecular movement and increases the extraction amount, but when the temperature exceeds the certain temperature (35, 40 ℃), the structure of the protein may be affected, the protein may be precipitated from the solution, and the extraction amount may be decreased. Therefore, albumin was selected for response surface analysis: 30. 35 and 40 ℃; globulin: 30. 35 and 40 ℃; prolamin: 30. gluten at 35, 40 ℃: 35. suitable temperatures are 40 ℃ and 45 ℃.
2.2.3 Effect of extraction time on protein content of Components
As can be seen from FIG. 4, when the time is 20-60 min, the extraction amount of albumin and prolamin increases, and reaches the maximum of 4.49mg/g and 2.49mg/g at 40 min. The extraction of gluten and globulin reached maximum values of 20.44mg/g, 9.26mg/g when the time was 20 min. When the extraction time exceeds 40min (albumin, prolamin) and 20min (gluten, globulin), the extraction amount of the four components is gradually reduced. This may result in substantial dissolution of the four proteins of rice at 40min and 20min, continued increase in extraction time, partial precipitation of the proteins, and reduced extraction yield. Therefore, albumin was selected for response surface analysis: 20. 40, 60 min; globulin: 10. 20, 30 min; prolamin: 20. 40, 60min gluten: 10. 20 and 30min are suitable time.
2.2.4 Effect of extraction times on protein content of Components
As can be seen from FIG. 5, the amount of albumin, globulin and gluten extracted increases when the number of extraction times is 2-4, and reaches the maximum values of 4.59mg/g, 10.29mg/g and 21.79mg/g when the number of extraction times is 3. When the extraction times are 2 times, the alcohol content reaches a maximum of 1.70 mg/g. When the number of extractions exceeded 3 (albumin, globulin, gluten) and 2 (alcoprotein), the amount of protein extraction of the four components gradually decreased. This may cause that four proteins of rice are basically dissolved out when the extraction is carried out for 3 times and 2 times, and as the extraction frequency continues to increase, the dissolution amount of other substances increases, so that partial precipitation of proteins in the final sample liquid occurs, and the extraction amount is reduced. Therefore, albumin was selected for response surface analysis: 2.3, 4 times; globulin: 2.3, 4 times; prolamin: 1.2, 3 times gluten: 2. the number of extraction times is preferably 3 or 4.
2.3 response surface test results and analysis
2.3.1 response surface test design and results
Based on the optimal conditions to be determined by the single-factor test results, the extraction conditions corresponding to each protein component in the rice are as follows: the feed-liquid ratio (A), the extraction temperature (B), the extraction time (C) and the extraction times (D) are independent variables, the response value (the content of the component protein) is a dependent variable, 4-
albumin: y is 1.5813A +0.7815B +0.0055C +3.1192D-0.0126AB + 0.0041 AC-0.0615AD +0.0019BC +0.0120BD +0.0044CD-0.0272A2-0.0010B2- 0.0020C2-0.3854D2-29.9904
Globulin: Y-0.0608A-0.6058B +0.2492C +0.0892D +0.2000 AB-0.3350 AC +0.2475AD +0.0200BC+0.4075BD+0.2825CD-0.5525A2-0.8350B2- 0.7850C2-0.8250D2+10.18
Prolamin:
Y＝0.1575-0.0192B-0.0117C+0.005D-0.1200AB-05075AC+0.0850 AD+0.0375BC+0.055BD+3600CD-0.5666A2-0.2516B2-0.4054C2-0.3229 D2+2.33
gluten:
Y＝0.6967A-0.3917B+0.4542C+0.6792D-0.0025AB+0.7450AC-0.3 025AD+0.7900BC+0.1225BD-0.0275CD-2.47A2-0.186B2-1.15C2-2.03D2+23.10
TABLE 2 response surface analysis protocol
2.3.2 analysis of variance and significance testing of regression models
Analysis of variance (ANOVA) was performed on the regression model using Design-Expert 8.0 software (tables 3, 4, 5), to determine the degree of influence of each factor on the extraction of rice component protein, and to examine the misformed lines of the regression equation. As can be seen from tables 3, 4 and 5, the model obtained by RSM four-component protein is very significant (P)<0.01), the mismatching terms have P values of 0.1038 (albumin), 0.3441 (globulin), 0.1234 (prolamin) and 0.2718 (glutelin), the P values are all more than 0.05, and the mismatching is not obvious; coefficient of correlation R2The model fit was better as indicated by 0.8969 (albumin), 0.8374 (globulin), 0.8569 (prolamin) and 0.9000 (gluten). Therefore, the established mathematical model is shown to successfully fit the test data, and the protein extraction amount of each component in the rice is well predicted within a certain range.
TABLE 3 Albumin response surface regression model analysis of variance
According to the significance analysis of the factors on the protein content of each component of rice, albumin: A. AC, B, C, D, AD, A2、B2、C2、D2The P value of (A) is less than 0.01, the P value of AB is less than 0.05, and according to the F value, the influence of four factors on the extraction amount of albumin is C>A>B>D; globulin: B. a. the2、B2、C2、D2The P value of (A) is less than 0.01, the P value of C, BD is less than 0.05, and the influence of four factors on the extraction amount of globulin is B according to the F value>C>D>A, and D, A did not reach a significant level of effect on globulin extraction, indicating that D, A had less effect on globulin extraction than B, C; prolamin: A. CD, A2、B2、C2、D2The P value of (A) is less than 0.01, and the influence of four factors on the extraction amount of globulin can be obtained according to the F value>B>C>D, and D, B, C did not reach a significant level in alcohol-soluble protein extraction, indicating that D, B, C had less effect on alcohol-soluble protein extraction than a; gluten: A. d, A2、B2、C2、D2The P value of (A) is less than 0.01, the P values of AC, BC, B and C are less than 0.05, and the influence of the four factors on the extraction amount of the gluten is A according to the F value>D>C>B. Significance analysis of each model also showed that the effect of each factor on the protein content of the extracted component was not generally linear.
TABLE 4 prolamin response surface regression model analysis of variance
TABLE 5 analysis of variance of the gluten response surface regression model
2.3.3 response surface analysis
According to the test result, drawing is carried out by using Design-Expert11 software, response surfaces (figure 6, figure 7, figure 8 and figure 9) of four component proteins under different extraction conditions and the interaction of the four factors can be obtained, and the interaction of the factors can be intuitively reflected. The gentler the response surface, the less sensitive the response value to the change of the factor, and conversely, the steeper the response surface, the more sensitive the response value to the change of the factor. Albumin: the interaction surface of the feed-liquid ratio, the extraction temperature, the extraction times and the extraction time is steep, the interaction is obvious (figure 6), the extraction temperature, the extraction time, the extraction times and the interaction surface are mild, the interaction is not obvious, the extraction time and the extraction times and the interaction surface are also mild, the interaction is not obvious, and the result is consistent with the analysis result of variance. Globulin: the extraction temperature and the extraction frequency are high, the interaction surface is steep, the interaction is obvious, the interaction surface among other factors is mild, and the interaction is not obvious. Consistent with the anova results (fig. 7). Prolamin: the interactive surface of the extraction time and the extraction times is steep, the interaction is obvious, the interactive surface of other items is mild, the interaction is not obvious, and the result is consistent with the analysis result of variance (figure 8). Gluten: the interaction surface of the feed-liquid ratio and the extraction time, the extraction temperature and the extraction time, and the interaction surface of the extraction time and the extraction frequency are steep, the interaction is obvious, the interaction surfaces of other items are mild, the interaction is not obvious, and the result is consistent with the analysis result of variance (figure 9). The response value gradually increases as the factor values increase, but gradually decreases as the factor values continue to increase after the response value reaches the maximum value.
2.3.4 validation test
According to the model established by the test, the optimal conditions for extracting the proteins of each component are as follows: the liquid-material ratio is 1:21.64, the extraction temperature is 31.57 ℃, the extraction time is 43.42min, the extraction times are 3.06 times, and the ratio of globulin: the liquid-material ratio is 1:19.65, the extraction temperature is 33.1 ℃, the extraction time is 21.6min, the extraction times are 2.97 times, and the alcohol soluble protein: the liquid-material ratio is 1:20.75, the extraction temperature is 34.63 ℃, the extraction time is 21.6min, the extraction times are 2.01 times, and the ratio of gluten: the material ratio is 1:15.83, the extraction temperature is 34.7 ℃, the extraction time is 22.35min, and the extraction times are 3.15. The extraction amount of each component protein under the condition is respectively 4.34mg/g, 10.31mg/g, 2.34mg/g and 23.27 mg/g.
Considering the feasibility of the experimental procedure, the optimized extraction conditions were modified to be, albumin: the ratio of feed to liquid is 1:22, the extraction temperature is 31.6 ℃, the extraction time is 43min, the extraction times are 3 times, and the ratio of globulin: the ratio of material to liquid is 1:20, the extraction temperature is 33.1 ℃, the extraction time is 22min, the extraction times are 3 times, and the alcohol soluble protein: the ratio of feed to liquid is 1:21, the extraction temperature is 34.6 ℃, the extraction time is 40min, the extraction times are 2 times, and the ratio of gluten: the ratio of material to liquid is 1:16, the extraction temperature is 34.7 ℃, the extraction time is 22min, and the extraction times are 3 times. Under these conditions, 3 repeated tests were carried out, the average extraction amount of each component protein of rice, albumin: 4.26mg/g, globulin: 9.76mg/g, alcolsoluble protein: 2.27mg/g, gluten: 23.60mg/g, and the relative error with the predicted value is 0.0187 (albumin), 0.0563 (globulin), 0.0308 (prolamin) and 0.014 (gluten), which shows that the optimal extraction process condition obtained by the optimization of the response surface method is better and has application value.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
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