CN103976134B - The method that high-strength ultrasonic improves Soybean Protein Isolate Gel intensity in conjunction with the oxidation of hydroxyl radical free radical appropriateness - Google Patents
The method that high-strength ultrasonic improves Soybean Protein Isolate Gel intensity in conjunction with the oxidation of hydroxyl radical free radical appropriateness Download PDFInfo
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- CN103976134B CN103976134B CN201410215909.9A CN201410215909A CN103976134B CN 103976134 B CN103976134 B CN 103976134B CN 201410215909 A CN201410215909 A CN 201410215909A CN 103976134 B CN103976134 B CN 103976134B
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Abstract
The invention discloses a kind of method that high-strength ultrasonic improves Soybean Protein Isolate Gel intensity in conjunction with the oxidation of hydroxyl radical free radical appropriateness, described method is: at FeCl3, Asc and H2O2Oxidation system adding soybean protein isolate, in high-strength ultrasonic, then carries out oxidation cultivate, making soybean protein isolate that oxidation in various degree to occur, thus improving its gel strength.Test result indicate that, in oxidation system, the concentration of soybean protein isolate is 30mg/mL, H2O2Concentration is 10mmol/L, and output is 600W/cm2, frequency be 20kHz high-strength ultrasonic in the gel strength of the oxidation obtained soybean protein isolate of 1h best, its gel strength is 2.35N, improves 176.47% compared with blank group.In the present invention, application high-strength ultrasonic is in conjunction with hydroxyl radical free radical appropriateness oxidation technology, it is possible to produce the soybean protein isolate with high-gel strength.
Description
Technical field
The present invention relates to a kind of method improving Soybean Protein Isolate Gel intensity, be specifically related to a kind of method that high-strength ultrasonic improves Soybean Protein Isolate Gel intensity in conjunction with the oxidation of hydroxyl radical free radical appropriateness.
Background technology
Soybean protein isolate (SoybeanProteinIsolate, SPI) it is with low denatured defatted soybean meal for raw material, a kind of protein content that the process technology of employing advanced person the is produced Functional Soybean Protein Isolate matter goods up to more than 90%, soybean protein isolate is after high-speed low temperature is centrifugal, 2S, 7S, 11S and 15S totally 4 grades can be divided into by sedimentation index, most important of which is that 7S β-companion's globulin (β-conglycinin) and 11S glycinin (glycinin).7S is three subunit (α1, α and β) different combine the trimers formed, they are combined by hydrophobic bond and hydrogen bond.Each 7S contains a small amount of disulfide bond, and without sulfydryl.11S is made up of 6 subunits, and each subunit is formed AB subunit by an Acid polypeptide chain (A) and a basic polypeptide chain (B) by a disulfide bond connection.11S molecule contains more disulfide bond and has sulfydryl.
SPI is the major protein batch materials in food industry, has material impact for improving the texture characteristic of processed food, improving trophism.But, different types of processed food is different for the requirement of required SPI dispensing functional characteristic.But, the SPI goods of the current industrialization of China are primarily present that product specific aim is strong, kind is few, the functional series bottleneck problem such as single, and the performance of product can not fully meet the demand of modern food processing.
SPI is applied in food system, can form more stable gel after heating, has higher glutinous bullet degree and plasticity simultaneously, and the processing of food is extremely advantageous.Meanwhile, the gel process of SPI is a sufficiently complex process, generally includes molecular stretching, several process such as division-combination and gathering etc..Protein forms what gel had been cross-linked with each other mainly by wide-spread peptide chain.In thermal denaturation process, natural SPI conformation is fully extended thus exposing some functional groups (such as sulfydryl and hydrophobic group).In order to reduce the energy of system, these groups exposed are combined by disulfide bond and hydrophobic interaction and form three dimensional network structure.When SPI concentration is more than or equal to the concentration of gel critical point, form gel networks.
It addition, SPI is similar with other compositions such as lipid and pigment, in processing and storage, it is easily subject to oxidative attacks.Protein oxidation is that directly induced by active oxygen or caused by indirectly reacting with lipid peroxide structural modification.At present, existing domestic and international correlational study shows slight or appropriate oxidation, it is possible to increase the functional characteristic of SPI.But all it is concentrated mainly in lipid free radical (such as AAPH, MDA, HPODE etc.) about the SPI research aoxidized, and the research about hydroxy radical appropriateness oxidation raising SPI gel strength there is no.
Meanwhile, high-strength ultrasonic (HighIntensityUltrasound, HIUS) modification technology is a kind of emerging, safety, nontoxic and environmental protection protein modified technology.The frequency range of HIUS is at 16-100kHz, and power can reach 1000W/cm2.The void effect that HIUS produces can interrupt quaternary structure of protein, discharges little molecule subunit or peptide, therefore, it is possible to significantly improve the functional characteristic (mainly dissolubility, emulsibility, gelation etc.) of SPI.And, HIUS can speed up the process of induced oxidation reaction.
Summary of the invention
It is an object of the invention to provide a kind of high-strength ultrasonic of applying and aoxidize the method improving Soybean Protein Isolate Gel intensity in conjunction with hydroxyl radical free radical appropriateness, the method has positive promotion impact for the gel strength of soybean protein isolate.
It is an object of the invention to be achieved through the following technical solutions:
At FeCl3, Ascorbate (Asc) and H2O2Adding concentration in oxidation system is the soybean protein isolate of 10～40mg/mL, FeCl in control system3The concentration that concentration is 0.1mmol/L, Asc be 0.1mmol/L, H2O2Concentration range be 0.1～15mmol/L, soybean protein isolate concentration be 10～40mg/mL, being then placed in output is 400～800W/cm2, frequency be 5～25kHz high-strength ultrasonic in oxidation cultivation 1～5h, make soybean protein isolate that oxidation in various degree to occur, thus improving its gel strength.
According to result of the test it is presumed that the oxidation of appropriateness (i.e. hydroxyl radical free radical oxidative system, referred to as HRGS.Main by FeCl3, Ascorbate and H2O2Reactive oxygen free radical is produced by the redox reaction of ferrum) albumen of expansion can be made to expose more hydrophobic group, thus forming solvable aggregation, the time of this concentration depending primarily on oxidant and oxidation.Meanwhile, high-strength ultrasonic processes and can fully be come out by intramolecular for soybean protein isolate hydrophobic region, adds the surface hydrophobic of soybean protein isolate.Owing to high-strength ultrasonic can form substantial amounts of soluble aggregate in conjunction with after hydroxyl radical free radical appropriateness oxidation technology processed soybeans separation albumen, therefore the soybean protein isolate after applying modified is when preparing gel, the hardness of gel is main relevant with two aspect factors, i.e. disulfide bond content and aggregate size.The soluble aggregate size formed during appropriateness oxidation is moderate, is evenly distributed, and the gel network of formation is dense, thus being conducive to improving gel strength.In oxidation system, the concentration of soybean protein isolate is 30mg/mL, H2O2Concentration is 10mmol/L, and output is 600W/cm2, frequency be 20kHz high-strength ultrasonic in the gel strength of the oxidation obtained soybean protein isolate of 1h best, its gel strength is 2.35N, improves 176.47% compared with blank group.In the present invention, application high-strength ultrasonic is in conjunction with hydroxyl radical free radical appropriateness oxidation technology, it is possible to produce the soybean protein isolate with high-gel strength.
Accompanying drawing explanation
Fig. 1 is that high-strength ultrasonic aoxidizes the impact on Soybean Protein Isolate Gel intensity in conjunction with hydroxyl radical free radical,A-GIn same string letter, identical then expression difference notable (P > 0.05), difference then represents significant difference (P < 0.05).
Detailed description of the invention
Below in conjunction with accompanying drawing, technical scheme is further described; but it is not limited thereto; every technical solution of the present invention modified or equivalent replaces, without deviating from the spirit and scope of technical solution of the present invention, all should be encompassed in protection scope of the present invention.
A kind of method that high-strength ultrasonic improves Soybean Protein Isolate Gel intensity in conjunction with the oxidation of hydroxyl radical free radical appropriateness, specifically comprises the following steps that
1, materials and methods
1.1 materials
Ferric chloride, ethylenediaminetetraacetic acid (EDTA), hydrogen peroxide, ascorbic acid, disodium hydrogen phosphate, sodium dihydrogen phosphate.
The extraction of 1.2 soybean protein isolates
Extracting degreasing low temperature soy meal powder (offer of Xiang He oils and fats company limited of Heilongjiang Province) mixes by the deionized water of 200g and 15 times, with 2mol/LNaOH tune pH value to 7.5-8.0, after stirring 1h, by its suspension centrifugal 20min of 8000g under 4 DEG C of conditions, take supernatant 2mol/LHCl and adjust pH to 4.5.After standing, under 4 DEG C of conditions, 8000g is centrifuged 10min.Take albumen precipitation be scattered in water and adjust pH to 7.0 with 2mol/LNaOH.Lyophilization is placed on 4 DEG C and saves backup.
The preparation of 1.3 hydroxyl radical free radical oxidation systems
By FeCl3, Ascorbate and H2O2Reactive oxygen free radical is produced, i.e. hydroxyl radical free radical oxidative system (Ahydroxylradical-generatingsystem, referred to as HRGS) by the redox reaction of ferrum.FeCl in fixed system3With Asc concentration, change H2O2Concentration；I.e. 0.1mmol/LFeCl3And 0.1mmol/LAsc, H2O2Concentration selects 0.1 respectively, 0.5,1,5,10,15mmol/L.
1.4 high-strength ultrasonics are in conjunction with hydroxyl radical free radical oxidation processes soybean protein isolate
Above-mentioned oxidation system adds soybean protein isolate so that the ultimate density of albumen is 10～40mg/mL.Then at room temperature (20 DEG C) put into output is 400～800W/cm2, frequency be 5～25kHz high-strength ultrasonic in oxidation cultivation 1～5h.Blocking reaction is carried out by adding BHA/Trolox/EDTA (making its ultimate density is 1mmol/L).In order to reduce the oxidising agent impact on testing index, oxidation product to remove supernatant through the washing of pH6.0 phosphate buffer and centrifugal treating (10000g/5min), then obtains, then through frozen dried, the oxidation protein that we need.Soybean protein isolate after oxidation measures its protein content by biuret method, is then passed through lyophilization and is placed on 4 DEG C and saves backup.
The preparation of 1.5 Soybean Protein Isolate Gels
Weigh the soybean protein isolate of above-mentioned different degree of oxidation, preparation protein concentration is to 12% (w/w) and is loaded on the beaker of 25mL, beaker is placed in the water-bath of 90 DEG C and is incubated 30min after covering preservative film by vacuum outgas, and ice bath preserves 24h after being cooled to room temperature in the refrigerator of 4 DEG C, to be measured.
The mensuration of 1.6 gel strengths
From refrigerator, take out gel at room temperature (20 DEG C) the ageing 30min prepared can measure.It is placed in testing sample on mensuration platform to fix, under room temperature, utilizes food texture measurement to measure.Measuring the intensity of gel with matter structure profile analysis method (Textureprofileanalyse writes a Chinese character in simplified form TPA), select the cylindric tack drift of diameter 10mm, set pace as 2mm/s, drawing velocity is 1mm/s；Recession speed is 2mm/s, and press depth is 10mm, once pops one's head in down in mensuration process and presses twice, and each sample measures in triplicate, and hardness (gel strength) is peak-peak during first time compression.
2, result
We it has been determined that appropriate oxidizing condition basis on, soybean protein isolate is processed by combined high-strength ultrasound wave, and by data above is analyzed, as shown in Figure 1, in oxidation system, the concentration of soybean protein isolate is 30mg/mL, H2O2Concentration is 10mmol/L, and output is 600W/cm2, frequency be 20kHz high-strength ultrasonic in the gel strength of the oxidation obtained soybean protein isolate of 1h the highest, its gel strength is 2.35N, improves 176.47% compared with blank group.
Claims (6)
1. the method that a high-strength ultrasonic improves Soybean Protein Isolate Gel intensity in conjunction with the oxidation of hydroxyl radical free radical appropriateness, it is characterised in that described method step is as follows: at FeCl3, ascorbic acid and H2O2Oxidation system adds soybean protein isolate, in high-strength ultrasonic, then carries out oxidation cultivate, make soybean protein isolate that oxidation in various degree to occur, thus improving its gel strength, FeCl in described oxidation system3Concentration be 0.1mmol/L, ascorbic acid concentration be 0.1mmol/L, H2O2Concentration be 0.1～15mmol/L, soybean protein isolate concentration be 10～40mg/mL, described oxidation incubation time is 1～5h, and described oxidation reaction is that the BHA/Trolox/EDTA of 1mmol/L stops by adding ultimate density.
2. the method that high-strength ultrasonic according to claim 1 improves Soybean Protein Isolate Gel intensity in conjunction with the oxidation of hydroxyl radical free radical appropriateness, it is characterised in that described ultrasonic power output is 400～800W/cm2, frequency be 5～25kHz.
3. the method that high-strength ultrasonic according to claim 1 improves Soybean Protein Isolate Gel intensity in conjunction with the oxidation of hydroxyl radical free radical appropriateness, it is characterised in that described soybean protein isolate concentration is 30mg/mL.
4. the method that high-strength ultrasonic according to claim 1 improves Soybean Protein Isolate Gel intensity in conjunction with the oxidation of hydroxyl radical free radical appropriateness, it is characterised in that described H2O2Concentration is 10mmol/L.
5. the method that high-strength ultrasonic according to claim 2 improves Soybean Protein Isolate Gel intensity in conjunction with the oxidation of hydroxyl radical free radical appropriateness, it is characterised in that described output is 600W/cm2, frequency be 20kHz.
6. the method that high-strength ultrasonic according to claim 1 improves Soybean Protein Isolate Gel intensity in conjunction with the oxidation of hydroxyl radical free radical appropriateness, it is characterised in that described oxidization time is 1h.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101569379A (en) * | 2009-06-02 | 2009-11-04 | 东北农业大学 | Method for preparing high gel stability soybean protein isolate |
| CN102934731A (en) * | 2012-12-07 | 2013-02-20 | 东北农业大学 | Preparation method of high-gelation soybean protein |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101569379A (en) * | 2009-06-02 | 2009-11-04 | 东北农业大学 | Method for preparing high gel stability soybean protein isolate |
| CN102934731A (en) * | 2012-12-07 | 2013-02-20 | 东北农业大学 | Preparation method of high-gelation soybean protein |
Non-Patent Citations (3)
| Title |
|---|
| Chemical，Physical，and Gel-forming Properties of Oxidized Myofibrils and Whey-and Soy-protein Isolates;Liu G, Xiong Y L, Butterfield D A;《Food Chemistry and Toxicology》;20001231;第65卷(第5期);第811页摘要、第815页第二栏第2段及第三栏第1段、第816页第三栏最后一段、第817页第一栏第1段 * |
| 羟基自由基氧化对乳清蛋白凝胶流变性质的影响;崔旭海;《食品与发酵工业》;20101231;第36卷(第3期);第40-45页 * |
| 超声处理对大豆分离蛋白热致凝胶功能性质的影响;朱建华;《食品与生物技术学报》;20060131;第25卷(第1期);第15页摘要、第16页第1.2小节、第17-18页第2.2、2.3小节 * |
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