CN111149912A - Modified soybean protein powder and preparation method thereof - Google Patents
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/14—Vegetable proteins
- A23J3/16—Vegetable proteins from soybean
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Abstract
The invention relates to a modified soybean protein powder and a preparation method thereof, wherein the preparation method comprises the following steps: dissolving soy protein in pure water to obtain a soy protein solution; treating the soybean protein solution by using atmospheric pressure cold plasma to obtain a modified soybean protein solution; and sequentially concentrating and drying the modified soybean protein solution to obtain the modified soybean protein powder. Therefore, the space structure and conformation epitope of the soybean protein are changed by utilizing the interaction between the active particles in the cold plasma and the soybean protein, and the aim of improving the processing functional characteristic of the soybean protein and reducing the allergenicity of the soybean protein is fulfilled. The invention has simple preparation, quick effect, low equipment investment, no use of any chemical organic reagent, good product quality and good safety.
Description
Technical Field
The invention relates to the field of food, and in particular relates to modified soybean protein powder and a preparation method thereof.
Background
The soybean protein has the characteristics of high nutritive value, easy preparation, wide source, low cost and the like, and is one of important raw materials in the fields of food processing and pharmacy. However, the natural functional properties of soy protein do not meet the needs of modern industrial products, which limits the practical application of soy protein to some extent. In addition, soybean is one of easily sensitized foods recognized by grain and agriculture organizations of the united nations. The incidence of soybean allergy in infants is about 0.4%, and clinical symptoms include rhinitis, rash, asthma, gastrointestinal discomfort and the like, which may affect the growth and development of infants in severe cases. Therefore, the method has important practical significance and wide market prospect for improving the functional characteristics of the soybean protein and reducing the sensitization of the soybean protein.
Currently, methods for modifying soy protein are commonly used, both in the hot and non-hot processing modes. Compared with the defects that the heat processing easily causes protein aggregation, denaturation and nutrient loss, the non-heat processing is adopted to treat the soybean protein at present. The existing non-thermal processing methods include hydrolysis, cross-linking, high voltage, ultrasound, pulsed electric field, radiation, etc. However, the treatment method of the related art only improves one of the processing functional properties and sensitization of the soybean protein, but loses or does not improve the other property when the improvement of one property is achieved, and it is difficult to achieve both of them.
Disclosure of Invention
The present invention is directed to solving, at least to some extent, one of the technical problems in the related art. Therefore, the invention aims to provide a preparation method of modified soybean protein powder. The method can improve soybean protein processing function, and reduce sensitization of soybean protein.
The second purpose of the invention is to provide modified soybean protein powder.
Therefore, in one aspect of the present invention, the present invention provides a method for preparing modified soy protein powder, which comprises the following steps:
(1) dissolving soy protein in pure water to obtain a soy protein solution;
(2) treating the soybean protein solution by using atmospheric pressure cold plasma to obtain a modified soybean protein solution;
(3) and sequentially concentrating and drying the modified soybean protein solution to obtain the modified soybean protein powder.
According to the preparation method of the modified soybean protein powder, the soybean protein is treated by the atmospheric pressure cold plasma, so that active particles in the cold plasma and the soybean protein are subjected to chemical interaction, the conformational structure and the sensitization epitope of the soybean protein are changed, and the purposes of simultaneously improving the processing functional characteristics and antigenicity of the soybean protein such as solubility, emulsibility and foamability are achieved. The method is simple and convenient to operate, quick in effect taking and remarkable in improvement effect; meanwhile, no organic chemical reagent is used, and the prepared soybean protein has excellent quality and good safety and is easy to realize industrialized production.
In addition, the preparation method of the modified soybean protein powder provided by the embodiment of the invention can also have the following additional technical characteristics:
according to an embodiment of the invention, the carrier gas of the atmospheric pressure cold plasma is air.
According to an embodiment of the invention, the conditions of the atmospheric pressure cold plasma treatment are: the processing voltage is 40kV, the processing frequency is 80-120 Hz, and the processing time is 2-10 min.
According to an embodiment of the present invention, the soy protein is soy protein isolate.
According to an embodiment of the present invention, in step (1), the soy protein is dissolved in pure water at a concentration of 20 mg/mL.
According to an embodiment of the present invention, in the step (3), the modified soy protein solution is concentrated by ultrafiltration and then spray-dried or freeze-dried.
The invention provides modified soybean protein powder on the other hand, which is prepared by adopting the preparation method of the modified soybean protein powder.
According to the modified soybean protein powder, the soybean protein with high processing function and low sensitization can be prepared by the method.
Additional aspects and advantages of the invention will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the invention.
Drawings
FIG. 1 is a functional property of processing of soy protein according to an embodiment of the present invention;
FIG. 2 is a simultaneous fluorescence spectrum of soy protein according to an embodiment of the present invention;
FIG. 3 is a circular dichroism spectrum of soy protein according to an embodiment of the present invention;
FIG. 4 is a secondary structure composition of soy protein according to an embodiment of the present invention;
FIG. 5 is a graph showing the IgE binding ability of soy proteins according to examples of the present invention.
Detailed Description
The technical solution of the present invention is illustrated by specific examples below. It is to be understood that one or more method steps mentioned in the present invention do not exclude the presence of other method steps before or after the combination step or that other method steps may be inserted between the explicitly mentioned steps; it should also be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Moreover, unless otherwise indicated, the numbering of the various method steps is merely a convenient tool for identifying the various method steps, and is not intended to limit the order in which the method steps are arranged or the scope of the invention in which the invention may be practiced, and changes or modifications in the relative relationship may be made without substantially changing the technical content.
In order to better understand the above technical solutions, exemplary embodiments of the present invention are described in more detail below. While exemplary embodiments of the invention have been shown, it should be understood that the invention may be embodied in various forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.
The room temperature of the invention is 25 ℃.
According to the embodiment of the invention, the invention provides a preparation method of modified soybean protein powder, which comprises the following steps:
(1) dissolving soy protein in pure water to obtain soy protein solution. According to an embodiment of the present invention, soy protein is dispersed in pure water at a concentration of 20mg/mL and stirred to dissolve sufficiently to obtain a uniform soy protein solution. Specifically, the soybean protein powder may be dispersed in pure water and stirred at room temperature for 2 hours to sufficiently dissolve the soybean protein powder. According to a preferred example of the present invention, the soy protein powder may be a soy protein isolate powder.
(2) And treating the soybean protein solution by using atmospheric pressure cold plasma to obtain a modified soybean protein solution. According to the specific embodiment of the invention, the soy protein solution is packaged in a flat dish and subjected to atmospheric pressure cold plasma treatment at room temperature. According to the specific embodiment of the invention, the atmospheric pressure cold plasma carrier gas is air, the treatment voltage is 40kV, the treatment frequency is 80-120 Hz, and the treatment time is 2-10 min.
(3) And sequentially concentrating and drying the modified soybean protein solution to obtain the modified soybean protein powder. According to the specific embodiment of the invention, the modified soybean protein solution is subjected to ultrafiltration concentration and then spray drying or freeze drying to prepare the modified soybean protein powder.
Therefore, the high-activity particles in the cold plasma and the soybean protein are subjected to chemical reaction to change the spatial structure and conformation epitope of the soybean protein, so that the aims of simultaneously improving the processing functional characteristics and the allergenicity of the soybean protein are fulfilled; the solubility of the soybean protein prepared by the method is improved by 61.4-152.3%, the emulsibility is improved by 6.0-39.1%, the foamability is improved by 12.6-58.0%, and the allergenicity is reduced by 29.4-76.0%. The method is convenient to operate, easy to realize industrial production, free of toxic and harmful reagents, and good in product quality and safety.
The invention will now be described with reference to specific examples, which are intended to be illustrative only and not to be limiting in any way.
Example 1
A preparation method of modified soybean protein powder comprises the following steps:
(1) 2000mg of soybean protein powder is taken, 100mL of pure water is added, and the mixture is stirred for 2 hours at room temperature to obtain a uniform soybean protein solution with the concentration of 20 mg/mL.
(2) And (2) subpackaging the soybean protein solution obtained in the step (1) into plates with the diameter of 90mm, horizontally placing 10mL of each plate in a tray, placing the tray between electrodes of an atmospheric pressure cold plasma instrument, and treating for 2min under the conditions of output voltage of 40kV and frequency of 80Hz by taking air as carrier gas to obtain the modified soybean protein solution.
(3) And (3) collecting the modified soybean protein solution obtained in the step (2), and performing ultrafiltration concentration and spray drying to obtain the modified soybean protein powder.
Example 2
A preparation method of modified soybean protein powder comprises the following steps:
(1) 1000mg of soybean protein powder is taken and 50mL of pure water is added, and the mixture is stirred for 2h at room temperature to obtain a uniform soybean protein solution with the concentration of 20 mg/mL.
(2) And (2) subpackaging the soybean protein solution obtained in the step (1) into 50mL centrifuge tubes, wherein each centrifuge tube is 20mL, placing the centrifuge tubes between electrodes of an atmospheric pressure cold plasma instrument in an open manner, and treating for 10min under the conditions of output voltage of 40kV and frequency of 100Hz by taking air as carrier gas to obtain the modified soybean protein solution.
(3) And (3) collecting the modified soybean protein solution obtained in the step (2), and performing ultrafiltration concentration and freeze drying to obtain the modified soybean protein powder.
Example 3
A preparation method of modified soybean protein powder comprises the following steps:
(1) 2000mg of soybean protein powder is taken, 100mL of pure water is added, and the mixture is stirred for 2 hours at room temperature to obtain a uniform soybean protein solution with the concentration of 20 mg/mL.
(2) And (2) subpackaging the soybean protein solution obtained in the step (1) into plates with the diameter of 200mm, horizontally placing 20mL of each plate in a tray, placing the tray between electrodes of an atmospheric pressure cold plasma instrument, and treating for 5min under the conditions of output voltage of 40kV and frequency of 120Hz by taking air as carrier gas to obtain the modified soybean protein solution.
(3) And (3) collecting the modified soybean protein solution obtained in the step (2), and performing ultrafiltration concentration and spray drying to obtain the modified soybean protein powder.
Test examples
The modified soybean protein powder prepared in the examples 1 to 3 was used as a test object to perform a performance test of soybean protein.
1. Solubility, emulsifying and foaming properties of soy protein varied:
(1) solubility of soy protein:
modified soy protein solutions obtained in examples 1 to 3 were prepared at a concentration of 0.5mg/mL, respectively, and the solutions were adjusted to pH 4.5 with untreated soy protein solution as a control, and shaken at room temperature for 1 hour. Then, the mixture was centrifuged at 3000g for 15min, and the supernatant was collected to determine the soluble protein content by BCA method.
Solubility (%) ═ protein content in supernatant/protein content in original solution × 100%.
As a result, as shown in FIG. 1, the solubility of the modified soybean protein of example 1 was increased by 112.9% as compared with that of the untreated soybean protein. The solubility of the modified soy protein of example 2 increased by 152.3% over the untreated soy protein. The solubility of the modified soy protein of example 3 increased by 61.4% over the untreated soy protein.
(2) Emulsifiability of soy protein:
the modified soy protein solutions obtained in examples 1 to 3 were prepared at a concentration of 5mg/mL, respectively, and the untreated soy protein solution was used as a control, and the respective solutions were mixed with sunflower seed oil at a ratio of 1: 3(v/v) and homogenizing for 1min at 16000rpm by a homogenizer. The samples were then immediately diluted 1000-fold with 1% SDS solution and their absorbance was measured at a wavelength of 500 nm.
Emulsifiability (m)2(2 × 2.303 × OD × DF)/(1000 × θ × L × C), where OD is the absorbance of the sample; DF is the dilution factor; theta is the oil phase proportion, and is taken as 0.25; l is the optical path and is taken to be 1 cm; c is protein concentration (g/mL).
As shown in fig. 1, the modified soy protein of example 1 showed a 39.1% increase in emulsifiability over the untreated soy protein. The modified soy protein of example 2 had a 12.0% increase in emulsifiability over the untreated soy protein. The modified soy protein of example 3 had a 6.0% increase in emulsifiability over the untreated soy protein.
(3) Foaming properties of soybean protein:
modified soy protein solutions obtained in examples 1 to 3 were prepared at a concentration of 5mg/mL, respectively, and the solutions were homogenized for 2min at 16000rpm in a graduated test tube using an untreated soy protein solution as a control.
Foaming (%) - (V)F0-VL)/VLX 100% where VF0Volume of bubble (mL), VLIs the volume of the initial solution (mL).
As a result, as shown in FIG. 1, the foaming property of the modified soybean protein of example 1 was increased by 58.0% as compared with that of the untreated soybean protein. The foaming properties of the modified soy protein of example 2 increased by 12.6% over the untreated soy protein. The foaming properties of the modified soy protein of example 3 increased by 12.8% over the untreated soy protein.
2. Spatial conformational change of soy protein
Spatial conformational changes of soy protein were analyzed by endogenous fluorescence spectroscopy:
modified soy protein solutions obtained in examples 1 to 3 were added to cuvettes at a concentration of 0.5mg/mL, respectively, and scanned at an excitation wavelength of 295nm and an emission wavelength of 300 to 400nm, using untreated soy protein solution as a control.
As shown in fig. 2, the endogenous fluorescence intensity of the modified soybean protein of example 1 is significantly reduced compared to the control group, and the maximum absorption wavelength is blue-shifted, which indicates that the oxidative fluorescence quenching occurs in the sensitive amino acids such as tyrosine and tryptophan, and the change in the hydrophobic microenvironment of the aromatic amino acid residues, i.e., the change in the spatial conformation of the protein. The endogenous fluorescence intensity of the modified soybean protein in example 2 is obviously reduced compared with that of the control group, but is increased compared with example 1, the maximum absorption wavelength is not blue-shifted obviously compared with example 1, and the frequency and time for increasing the cold plasma can further induce the unfolding of the protein conformation, expose the internal hydrophobic groups and further increase the fluorescence intensity. The endogenous fluorescence intensity of the modified soy protein of example 3 was significantly reduced compared to both the control and example 2, but similar to example 1, the degree of blue-shift of the maximum absorption wavelength was similar to example 2, which is due to the additive effect of key amino acid residue oxidative quenching and protein conceived structure change.
3. Secondary structure change of soy protein
Secondary structure change of soy protein by circular dichroism analysis:
and respectively adding the modified soybean protein solution with the concentration of 0.2mg/mL obtained in the examples 1-3 into a sample tank of a circular dichroism chromatograph, scanning at room temperature, wherein the wavelength range is 260-190 nm, the scanning speed is 100nm/min, the bandwidth is set to be 1.0nm, and a CONTIN method is adopted to perform spectrum decomposition on a sample spectrum band to obtain the percentage of a secondary structure.
Results as shown in fig. 3 and 4, the modified soy protein of example 1 had a reduced β 1-fold (2.0%) and β -turn (13.7%) content, and an increased α -helix (56.2%) and random curl (28.1%) content, as compared to the control (55.1% α -helix, 2.8% β -fold, 15.1% β 0-turn, 27.0% random curl), the modified soy protein of example 1 had a reduced α -helix (48.2%), β -fold (2.3%) and β -turn (14.1%) content, and an increased random curl content (35.3%). the modified soy protein of example 2 had a reduced α -helix (46.3%) content, while the modified soy protein of example 3 had a reduced β -fold (3.5%), β -fold (17.8%) and increased random curl content (32.4%) content, thus indicating that the secondary structure of the modified soy protein was changed under different cold conditions.
4. Soy protein allergenicity assay
The binding capacity of IgE of soy protein was measured by indirect ELISA to reflect changes in sensitization of soy protein:
the modified soy protein solutions obtained in examples 1 to 3 were applied to an ELISA plate at a concentration of 500. mu.g/mL, respectively, at a concentration of 100. mu.L per well, and the untreated soy protein solution was used as a control. 4 ℃ overnight. Cleaning solutionThe plate was washed 5 times, 5min each time, and patted dry. Adding 200 μ L of bovine serum albumin containing 5% per well, sealing, incubating at 37 deg.C for 2h, and washing the plate for 5 times; adding 100 mu L/hole of soybean allergy patient mixed serum diluted at a ratio of 1:30, incubating at 37 ℃ for 1h, and washing the plate for 5 times after the reaction is finished; adding 200 mu L/hole of biotin-labeled sheep anti-human IgE diluted by 1:5000, incubating at 37 ℃ for 2h, and washing the plate for 5 times; adding TMB 100 μ L/well, and developing at 37 deg.C in dark for 30 min; after color development, 2M H was added2SO4The reaction was stopped at 50. mu.L/well and the absorbance (OD value) was measured at a wavelength of 450 nm.
IgE binding capacity (%) ═ 1-ODSample (I)/ODControl) X 100%, wherein, ODSample (I)And ODControlOD values of the sample group and the control group are respectively,
as a result, as shown in FIG. 5, the allergenicity of the modified soybean protein of example 1 was reduced by 29.4% as compared with that of the untreated soybean protein. The allergenicity of the modified soy protein of example 2 was reduced by 50.3% compared to the untreated soy protein. The allergenicity of the modified soy protein of example 3 was reduced by 76.0% over the untreated soy protein.
In conclusion, the invention utilizes the interaction between active particles in the cold plasma and the soybean protein to change the spatial structure and conformation epitope of the soybean protein, and can achieve the purpose of simultaneously improving the processing functional characteristics and the sensitization of the soybean protein by controlling the fixed treatment time and the exposure frequency. The solubility of the soybean protein treated by the method is improved by 61.4-152.3%, the emulsibility is improved by 6.0-39.1%, the foamability is improved by 12.6-58.0%, and the allergenicity is reduced by 29.4-76.0%. The invention has simple operation, quick effect, low equipment investment, no need of using any chemical reagent and good product safety.
In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above should not be understood to necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, various embodiments or examples described in this specification can be combined and combined by those skilled in the art.
Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention, and that variations, modifications, substitutions and alterations can be made to the above embodiments by those of ordinary skill in the art within the scope of the present invention.
Claims (7)
1. The preparation method of the modified soybean protein powder is characterized by comprising the following steps:
(1) dissolving soy protein in pure water to obtain a soy protein solution;
(2) treating the soybean protein solution by using atmospheric pressure cold plasma to obtain a modified soybean protein solution;
(3) and sequentially concentrating and drying the modified soybean protein solution to obtain the modified soybean protein powder.
2. The method for preparing soybean protein powder according to claim 1, wherein the carrier gas of the atmospheric pressure cold plasma is air.
3. The method for preparing soybean protein powder according to claim 1, wherein the conditions of the atmospheric pressure cold plasma treatment are as follows: the processing voltage is 40kV, the processing frequency is 80-120 Hz, and the processing time is 2-10 min.
4. The method for preparing soybean protein powder according to claim 1, wherein the soybean protein is soybean protein isolate.
5. The method for preparing soybean protein powder according to claim 1, wherein the soybean protein is dissolved in pure water at a concentration of 20mg/mL in step (1).
6. The method for preparing soybean protein powder according to any one of claims 1 to 5, wherein in the step (3), the modified soybean protein solution is concentrated by ultrafiltration and then spray-dried or freeze-dried.
7. A modified soybean protein powder, which is characterized by being prepared by the preparation method of the modified soybean protein powder as claimed in any one of claims 1 to 6.
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| CN112042927A (en) * | 2020-08-26 | 2020-12-08 | 华南理工大学 | Method and system for preparing ovalbumin high-efficiency emulsifier based on low-temperature plasma |
| CN112237288A (en) * | 2020-09-15 | 2021-01-19 | 浙江工业大学 | Method for making soybean protein stick |
| CN114391598A (en) * | 2022-02-10 | 2022-04-26 | 上海我爱冰淇淋食品有限公司 | Ice cream and preparation method thereof |
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| WO2024124632A1 (en) * | 2022-12-13 | 2024-06-20 | 江苏大学 | Method for recovering proteins from minced-fish washing waste liquid on the basis of low-temperature plasma technique |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112042927A (en) * | 2020-08-26 | 2020-12-08 | 华南理工大学 | Method and system for preparing ovalbumin high-efficiency emulsifier based on low-temperature plasma |
| CN112237288A (en) * | 2020-09-15 | 2021-01-19 | 浙江工业大学 | Method for making soybean protein stick |
| CN114391598A (en) * | 2022-02-10 | 2022-04-26 | 上海我爱冰淇淋食品有限公司 | Ice cream and preparation method thereof |
| CN114853842A (en) * | 2022-05-18 | 2022-08-05 | 南京财经大学 | Method for continuously modifying isolated soy protein based on gas-conditioning-assisted low-temperature plasma |
| CN114853842B (en) * | 2022-05-18 | 2023-07-18 | 南京财经大学 | Method for continuously modifying isolated soy protein based on modified atmosphere auxiliary low-temperature plasma |
| WO2024124632A1 (en) * | 2022-12-13 | 2024-06-20 | 江苏大学 | Method for recovering proteins from minced-fish washing waste liquid on the basis of low-temperature plasma technique |
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