CN114343190A - Method for improving plant protein emulsifying performance by using quillaja bark extract - Google Patents
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Abstract
The invention provides a method for improving the emulsifying performance of plant protein by using a quillaja bark extract, which comprises the following steps: s1, mixing and hydrating plant protein and a natural soapbark extract by using water or a buffer solution and alcohol according to a proper proportion to obtain a mixed solution of the plant protein and the soapbark extract; s2, drying the mixed solution of the vegetable protein and the soapbark extract to obtain a vegetable protein-soapbark extract compound. The invention does not need complex protein modification, can obviously improve the protein emulsifying property only by adding a small amount of natural soapbark extract, has simple and convenient process operation, no reagent and pure plant base, and is easy to realize industrialized continuous production.
Description
Technical Field
The invention relates to the technical field of food processing, in particular to a method for improving the emulsifying performance of vegetable protein by using a quillaja bark extract.
Background
At present, the requirements of people on the quality of life are gradually improved, green, natural and healthy foods are pursued, and safe and nontoxic natural food additives gradually become the key points of research. The protein has good emulsifying property, and the plant seed protein has wide sources, low cost, complete amino acid types and rich nutrition, so the plant protein is an important resource for developing natural emulsifiers. At present, many vegetable proteins have been shown to have potential as emulsifiers, such as pea, lentil, soy and corn germ proteins. Among vegetable protein emulsifiers, soy protein isolate and modified soy protein isolate are currently being studied. The soybean protein is prepared from defatted soybean meal, can provide all essential amino acids meeting the requirements of human bodies, can be comparable with animal protein, and belongs to one of the best vegetable proteins. The soybean protein has the functional characteristics of good emulsifying property, water absorption, oil retention property, foamability and the like, so the soybean protein can be used as an additive for supplementing nutrition and regulating food structure and is widely applied to food systems such as infant food formulas, meat products, dairy products and the like. However, the denaturation of protein caused by the high-temperature desolventizing process of the defatted soybean meal limits the functional characteristics of the soybean protein and cannot meet the requirements of specific food processing, and the improvement of the emulsifying property of the soybean protein becomes the most fundamental and important problem to be solved urgently.
At present, common methods for improving the emulsifying performance of soy protein can be divided into three types: physical modification, chemical modification and enzymatic modification. Zhao Zhan Ming et al disclose a preparation method (CN101317623B) of emulsified soybean protein isolate, wherein low-temperature defatted soybean meal powder is used as raw material, after enzymolysis by alkalase alkaline protease under alkaline condition, bottom residue is removed by centrifugation, and the obtained soybean protein enzymolysis supernatant is homogenized, acid-precipitated and dried to obtain the emulsified soybean protein isolate. Zhuxiuqing and the like disclose a preparation method of soybean protein isolate with acid resistance and high emulsification performance and a product thereof (CN 104397318B). The soybean protein isolate with acid resistance and high emulsification performance is obtained by adding phytase for the first enzymolysis after ultrasonic pretreatment of soybean protein isolate water solution, then adding acid protease for the second enzymolysis, inactivating enzyme, cooling, concentrating and drying. Zhuxiuqing et al also discloses a preparation method (CN103583791A) of high-emulsification stability soybean protein, which utilizes ultrasonic wave combined with alkylation modification to improve the emulsification stability of the soybean protein. Wuhaibo et al disclose a method (CN104351463A) for improving the emulsifying property of isolated soybean protein under acidic condition, which comprises pretreating isolated soybean protein by extrusion, adding protease for enzymolysis, inactivating enzyme, cooling, concentrating, and drying to improve the emulsifying property of isolated soybean protein under acidic condition. Plum sun exposure et al disclose a processing method (CN105104706A) of soybean protein isolate with high emulsifying activity, which comprises mixing soybean protein isolate powder and maltodextrin in a high-efficiency mixer, placing the mixture in a dry reactor for reaction, naturally drying, and cutting the product into pieces to obtain the high emulsifying activity protein product. Populus plumipes et al discloses a preparation method (CN110583850A) of high-emulsibility soybean protein, which improves the emulsibility of the soybean protein through the complicated steps of soaking, grinding, leaching, shaking, acid precipitation, homogenization, enzymolysis, glycosylation, sterilization, freeze-drying and the like. The protein structure is rapidly unfolded through physical modes such as heating, micro-jet, high-intensity ultrasonic treatment and the like, the emulsifying property is improved through the exposure of hydrophobic groups, and although the safety is high and the action time is short, the energy consumption is high, the treatment cost is high and the modification effect is limited. The emulsifying property is improved by chemical modification in the modes of acid alkalization, acylation, phosphorylation and the like, and the method has the advantages of low cost, simple operation, obvious effect and the like to make up for the deficiency of physical modification, but introduces a large amount of reagents and has limited application. Glycosylation and enzyme methods become more common modification modes, but the enzyme sources are less, the varieties are limited, the price is high, and the emulsification property of the grafting products generated by glycosylation is not outstanding. Therefore, it is critical to develop a method for more rapidly and effectively improving the emulsifiability of soybean protein.
Disclosure of Invention
The technical problem to be solved is as follows: the invention aims to provide a method for remarkably improving the protein emulsifying performance by adding a small amount of natural quillaja bark extract without complex protein modification, and the method has the advantages of simple and convenient process operation, no reagent, pure plant base and easy realization of industrial continuous production.
The technical scheme is as follows: a preparation method of a compound for improving the emulsifying property of plant protein comprises the following steps:
s1, mixing and hydrating plant protein and a natural soapbark extract by using water, a buffer solution or alcohol according to a proper proportion to obtain a mixed solution of the plant protein and the soapbark extract;
s2, drying the mixed solution of the vegetable protein and the soapbark extract to obtain a vegetable protein-soapbark extract compound. Preferably, the mass ratio of the vegetable protein to the natural quillaja bark extract in the step S1 is 100: 0.1-50.
Preferably, the vegetable protein includes soybean protein, peanut protein, buckwheat protein, wheat protein, pea protein, cottonseed protein, rapeseed protein or potato protein.
Preferably, the soy protein is soy protein isolate or soy protein concentrate.
Preferably, the quillaja bark extract is a liquid extract or an extract as a dehydrated solid.
Preferably, the drying method in step S2 includes freeze drying, atmospheric drying, reduced pressure drying, spray drying or boiling drying.
Use of a compound for improving the emulsifying properties of a vegetable protein in a food system in which the emulsifying properties of the protein are to be applied.
Has the advantages that: the compound for improving the emulsifying property of the vegetable protein has the following advantages:
1. the invention utilizes the addition of the quillaja bark extract to improve the emulsifying property of the soybean protein, and the compound is formed only through the intermolecular interaction promoted by wet mixing, and the compound can be used in any food system needing to add high-emulsifying soybean protein. The method retains high-quality full-component protein, protects nutritional value of soybean protein, and the added Quillaja Saponaria Molina extract has multiple biological activities of resisting oxidation, relieving inflammation, and reducing blood lipid. The soybean protein emulsion can be used for preparing functional soybean protein ingredients, can also be used as a high-emulsification soybean protein additive to be applied to foods, health products and cosmetics;
2. the soybean protein and the quillaja bark extract related by the invention are both plant sources, and have sustainability, the addition amount range of the quillaja bark extract in the process of improving the emulsibility of the soybean protein is wide, and is from 0.1 percent to 20 percent (relative to the mass percent of the protein) of the quillaja bark extract, and the effect of improving the emulsibility of the soybean protein is obvious;
3. the method for improving the emulsifying property of the soybean protein is simple, has strong flexibility and can realize industrial production.
Drawings
FIG. 1 is an appearance of emulsions corresponding to examples 1-4 and comparative example 1;
FIG. 2 is a graph showing particle size distributions of emulsions according to examples 1 to 4 and comparative examples 1 to 2;
FIG. 3 is a graph of the viscosity of emulsions corresponding to examples 1-4 and comparative example 1;
FIG. 4 is a graph showing fluorescence spectra of solutions of examples 1 to 4 and comparative example 1;
FIG. 5 is a graph of the surface hydrophobicity versus the ratio of examples 1-4 and comparative example 1;
FIG. 6 is a graph of infrared analysis of examples 5 to 6 and comparative example 1;
FIG. 7 is a graph of thermal stability analysis of examples 5-6 and comparative example 1;
Detailed Description
The present invention is further illustrated by the following examples, which are only intended to illustrate the present invention and not to limit the scope of the present invention.
Example 1
A method for improving plant protein emulsifying performance by using Quillaja saponaria Molina extract comprises the following steps:
s1, dissolving soybean protein and a natural quillaja bark extract in a phosphate buffer solution with the pH value of 7.0, wherein the mass concentrations of the soybean protein and the quillaja bark extract in the buffer solution are 1 wt% and 0.05 wt%, respectively, stirring for 2 hours at room temperature, standing at 4 ℃ for overnight reaction, and fully hydrating to obtain a mixed solution of the soybean protein and the quillaja bark extract;
s2, carrying out freeze drying on the mixed solution of the soybean protein and the soapbark extract to obtain a soybean protein-soapbark extract compound.
Example 2
A method for improving plant protein emulsifying performance by using Quillaja saponaria Molina extract comprises the following steps:
s1, dissolving soybean protein and a natural quillaja bark extract in a phosphate buffer solution with the pH value of 7.0, wherein the mass concentrations of the soybean protein and the quillaja bark extract in the buffer solution are 1 wt% and 0.09 wt% respectively, stirring for 2 hours at room temperature, reacting overnight at 4 ℃, and fully hydrating to obtain a mixed solution of the soybean protein and the quillaja bark extract;
s2, carrying out freeze drying on the mixed solution of the soybean protein and the soapbark extract to obtain a soybean protein-soapbark extract compound.
Example 3
A method for improving plant protein emulsifying performance by using Quillaja saponaria Molina extract comprises the following steps:
s1, dissolving soybean protein and a natural quillaja bark extract in a phosphate buffer solution with the pH value of 7.0, wherein the mass concentrations of the soybean protein and the quillaja bark extract in the buffer solution are 1 wt% and 0.13 wt% respectively, stirring for 2 hours at room temperature, standing at 4 ℃ for overnight reaction, and fully hydrating to obtain a mixed solution of the soybean protein and the quillaja bark extract;
s2, carrying out freeze drying on the mixed solution of the soybean protein and the soapbark extract to obtain a soybean protein-soapbark extract compound.
Example 4
A method for improving plant protein emulsifying performance by using Quillaja saponaria Molina extract comprises the following steps:
s1, dissolving soybean protein and a natural quillaja bark extract in a phosphate buffer solution with the pH value of 7.0, wherein the mass concentrations of the soybean protein and the quillaja bark extract in the buffer solution are 1 wt% and 0.18 wt%, respectively, stirring for 2 hours at room temperature, standing at 4 ℃ for overnight reaction, and fully hydrating to obtain a mixed solution of the soybean protein and the quillaja bark extract;
s2, carrying out freeze drying on the mixed solution of the soybean protein and the soapbark extract to obtain a soybean protein-soapbark extract compound.
Example 5
A method for improving plant protein emulsifying performance by using Quillaja saponaria Molina extract comprises the following steps:
s1, dissolving soybean protein and a natural quillaja bark extract in a phosphate buffer solution with the pH value of 7.0, wherein the mass concentrations of the soybean protein and the quillaja bark extract in the buffer solution are 1 wt% and 0.25 wt%, respectively, stirring for 2h at room temperature, standing at 4 ℃ for overnight reaction, and fully hydrating to obtain a mixed solution of the soybean protein and the quillaja bark extract;
s2, carrying out freeze drying on the mixed solution of the soybean protein and the soapbark extract to obtain a soybean protein-soapbark extract compound.
Example 6
A method for improving plant protein emulsifying performance by using Quillaja saponaria Molina extract comprises the following steps:
s1, dissolving soybean protein and a natural quillaja bark extract in a phosphate buffer solution with the pH value of 7.0, wherein the mass concentrations of the soybean protein and the quillaja bark extract in the buffer solution are 1 wt% and 0.5 wt%, respectively, stirring for 2h at room temperature, standing at 4 ℃ for overnight reaction, and fully hydrating to obtain a mixed solution of the soybean protein and the quillaja bark extract;
s2, carrying out freeze drying on the mixed solution of the soybean protein and the soapbark extract to obtain a soybean protein-soapbark extract compound.
The emulsifying performance of the present invention uses the average particle size of the emulsion as a main evaluation index, and uses the viscosity of the emulsion as an auxiliary evaluation index by rheological analysis.
The method for measuring the average particle size of the emulsion comprises the following steps: the particle size of the emulsion droplets was measured using a Mastersizer 3000 laser particle sizer. The parameters are set as follows: the refractive index of the particles is 1.440, and the absorption rate of the particles is 0.002; the dispersant is water, and the refractive index of the dispersant is 1.330. The average size of the droplet size is characterized by D4,3, the volume mean diameter, D4,3 ∑ nidi4/∑ nidi3, where di is the diameter of the emulsion droplets and ni is the number of emulsion droplets.
The method for measuring the viscosity of the emulsion comprises the following steps: the apparent viscosity of the emulsion was measured using an HAAKE MARS 60 rheometer. Set the shear rate range to 0.1s-1To 100s-1The change in viscosity was recorded.
The appearance of the emulsions of comparative example 1 and examples 1-4 (as shown in fig. 1) prepared using a 1 wt% soy protein solution prepared in comparative example 1 and a 0.09% quillaia bark extract solution prepared in comparative example 2 and soy protein solutions containing quillaia bark extracts at different concentrations prepared in examples 1-4 was observed and it was found that the fluidity of the emulsions was enhanced and the strong plasticity of the soy protein was reduced by gradually adding the quillaia bark extract to the 1 wt% soy protein-based emulsion corresponding to comparative example 1.
The emulsion was prepared by dissolving the powders of comparative examples 1-2 and examples 1-4 in a buffer solution as 25 parts by weight of an aqueous phase, adding 75 parts by weight of soybean oil, and shearing at 24000r/min for 3min at high speed to form an emulsion in which soybean protein and quillaia saponaria extract are jointly stabilized, wherein the total emulsion system was 20g, the aqueous phase was 5g, the oil phase was 15g, and the mass of soybean protein and quillaia saponaria extract was converted to the concentration shown in Table 1.
TABLE 1
The prepared emulsion was subjected to particle size analysis (as shown in Table 1), and from the results of comparative example 1, D4,3 was 14.25 μm with the addition of 1 wt% of the soy protein-based emulsion. Whereas the 0.09 wt% quillaja bark extract based emulsion prepared from comparative example 2 had a D4,3 of 11.53 μm. It can be clearly seen that the gradual addition of quillaja bark extract (0.05-0.18 wt%) in comparative example 1 effectively reduced the average particle size of the 1 wt% soy protein based emulsion from 14.25 μm to 7.32 μm. Furthermore, as can be seen from the emulsion particle size distribution results shown in fig. 2, the particle size distribution of the soy protein-based emulsions with the quillaia saponaria extract added all shifted to the left, indicating that the particle size of the emulsions prepared in examples 1-4 decreased with increasing quillaia saponaria extract concentration. The added quillay extract can be self-assembled on an oil-water interface in a single form or can generate intermolecular interaction with soybean protein on the interface to form a stable interface film, and the flocculation and coalescence among adjacent emulsion droplets are prevented by the acting forces such as electrostatic repulsion, steric hindrance and the like existing in molecules, so that the particle size of emulsion droplets is reduced, and the emulsifying property of the soybean protein can be improved by adding a small amount of quillay extract.
Rheological analysis of the emulsion revealed that (as shown in fig. 3) the viscosity of the 1 wt% soy protein based emulsion added with different quillaia bark extract concentrations gradually decreased with significant effect. Increasing the quillay extract concentration, meaning an increase in its interfacial adsorption, prevents and reduces the effective adsorption of proteins at the interface, thereby disrupting bridging flocculation that exists between soy proteins, resulting in a significant change in emulsion viscosity. In addition, the viscosity reduction of the emulsion is also related to the particle size of emulsion droplets, the particle size of the emulsion droplets is reduced, the resistance during shearing is reduced, and the viscosity reduction of the emulsion is more obvious. Such properties confirm that the development of high protein products can also be achieved with the addition of small amounts of quillaja bark extract.
In order to confirm that the soybean protein and the quillaja bark extract were combined to form a complex, the influence of the addition of the quillaja bark extract on the structure and properties of the soybean protein was studied by fluorescence spectroscopy, fourier infrared and differential scanning calorimetry, and the surface hydrophobicity of the soybean protein to which the quillaja bark extract was added was measured.
Fluorescence spectroscopy: comparative example 1 was prepared as a 1mg/g soybean protein solution, and examples 1 to 4 were prepared as soybean protein (1mg/g) solutions containing quillaja bark extract at concentrations of 0.5, 0.9, 1.3, and 1.8mg/g, respectively, and the temperature was maintained in a pan of constant temperature water bath at 25 ℃ for 5 min. Selecting an excitation wavelength of 280nm, excitation and emission slits of 5nm and a scanning speed of 12000nm/min, and scanning a fluorescence spectrum within the wavelength range of 300-500 nm in a 1cm quartz cuvette of an F-7000 fluorescence spectrometer.
Fourier infrared technology: the powders prepared in comparative example 1 and examples 5 to 6 were mixed with potassium bromide powder at a ratio of 1: 100(m/m), ground to uniformity, placed in a mold and tabletted at 32cm-1Under the condition of the resolution ratio of (1), scanning is carried out for 32 times, and the scanning distance is 4000-400 cm-1Infrared spectroscopy was scanned over the wavenumber range.
Differential scanning calorimetry: 2mg of the powder prepared in comparative examples 1-2 and examples 5-6 was weighed, placed in a sample cell of a differential scanning calorimeter, 20. mu.L of deionized water was added, a platen was sealed, and the mixture was allowed to equilibrate overnight at room temperature. The well-balanced sample cell is placed on the left side of the DPS console, and an empty box is used as a blank sample and placed on the right side. Temperature scanning range: 20-120 ℃; the heating rate is as follows: 10 ℃/min. The thermal transition parameters of the mixture were determined with data recording software.
Surface hydrophobicity of soybean protein: comparative example 1 and examples 1 to 4 were formulated into soybean protein solutions (0.1mg/g, 0.2mg/g, 0.3mg/g, 0.4mg/g, 0.5mg/g, 0.6mg/g) and 8.0 mmol/L1-anilino-8-naphthalenesulfonic Acid (ANS) solutions at different concentrations with a 20mM buffer (pH 7.0). Before detection, 20. mu.L of LANS solution was added to 4mL of protein solution, mixed well, and the fluorescence intensity of the mixture (FI1) and the fluorescence intensity of the sample without ANS solution (FI0) were measured quickly. The excitation wavelength was 370nm and the emission wavelength was 465 nm. The difference between FI1 and FI0 is marked as FI, the protein concentration is plotted on the abscissa and FI is plotted on the ordinate, and the slope of the curve is the surface hydrophobicity index H0 of the protein molecule.
The influence of the quillaja bark extract on the fluorescence spectrum of the soybean protein is researched by a fluorescence spectrum technology, and the result is shown in fig. 4, that the quillaja bark extract has an obvious quenching effect on the fluorescence of the soybean protein, and the fluorescence of the soybean protein is gradually quenched along with the increase of the mass concentration of the quillaja bark extract under the condition of a soybean protein solution with a certain mass concentration. The quillay extract red-shifts the maximum emission peak of soy protein from 337nm to 343nm, approximately 6 nm. This indicates that the quillaja bark extract and the soy protein may interact with each other, so that the spatial conformation of the soy protein isolate molecules is changed, and further the microenvironment around tryptophan is changed from a hydrophobic environment to a hydrophilic environment, and the peptide chain becomes more extended. Therefore, we determined that the surface hydrophobicity of soy protein decreased in a concentration-dependent manner as we expected (as shown in fig. 5), i.e., the addition of quillaia saponaria extract decreased the surface hydrophobicity of soy protein, and the decrease in the surface hydrophobicity of soy protein was more pronounced with the increase in the concentration of quillaia saponaria extract, consistent with the conclusion of fluorescence spectroscopy. This is because the hydrophobic aglycone present on the quillay extract is bound to the hydrophobic amino group of the soybean protein through hydrophobic interaction, resulting in the conversion of tryptophan to hydrophilic and the decrease of surface hydrophobicity.
To confirm the above hypothesis, the structures of comparative example 1 (soy protein) and examples 5-6 (complex formed by soy protein-quillaia saponaria extract) were analyzed by fourier infrared technique, and the results are shown in fig. 6. For proteins, fourier transform spectroscopy can provide information on the amino groups, the amide I band (or H-O-H bending vibration and C ═ O stretching vibration), the amide ii band (N-H bending), the amide iii band (or C-O and C-O-C vibration), the C-C vibration in the protein cyclic structure, and the C-O glycosidic bond vibrational band in the protein molecule. And (3) carrying out baseline correction, deconvolution processing and second derivative fitting processing on the infrared spectrum analysis of the protein in sequence, and ensuring the minimum fitting residual error through multiple fitting. Referring to the existing research, the corresponding relationship between each sub-peak and the secondary structure is as follows: 1615-1637 cm-1And 1682-1700 cm-1Is in beta-folded structure, 1646-1664 cm-1Is of an alpha-helical structure, 1637-1645 cm-1Is a random coil structure, 1664-1681 cm-1Is in a beta-corner structure. As the concentration of quillaia saponaria extract added increased, the beta-sheet content increased (18.18% to 26.93%), while the alpha-helix content (25.68% decreased to 22.32%), the beta-turn content (30.68% decreased to 28.21%) and the random coil content (25.46% decreased to 22.53%), indicating that the partial alpha-helix structure, the beta-turn structure and the random coil structure gradually transformed the beta-sheet structure. While beta-sheet structures are common in the internal folding regions of proteins, hydrophobic interactions are the main driving force for protein folding, and the increase in the content of beta-sheet structures indicates that quillaia saponaria bark extract can effectively bind to soy proteins through hydrophobic interactions.
Further, the thermal stability of comparative examples 1-2 and examples 5-6 was investigated by differential scanning calorimetry, and the results are shown in FIG. 7. As is clear from comparative example 1, the two denaturation temperatures of the soy protein used were 71.8 ℃ and 86.5 ℃ corresponding to the beta-conglycinin and glycinin fractions, respectively. Whereas the quillaia saponaria extract of comparative example 2 did not change at all in this temperature range. It is noted that the right shift of the peaks in examples 5 and 6 illustrates that the addition of quillaja bark extract can improve the thermal stability of soy protein, which is caused by the quillaja bark extract having strong hydrophobic aglycone binding to the hydrophobic amino group of soy protein through hydrophobic interaction to change the structure of soy protein, i.e., form a soy protein-quillaja bark extract complex.
Claims (7)
1. A method for improving the emulsifying performance of plant protein by using a quillaja bark extract is characterized by comprising the following steps:
s1, mixing and hydrating plant protein and a natural soapbark extract by using water, a buffer solution or alcohol according to a proper proportion to obtain a mixed solution of the plant protein and the soapbark extract;
s2, drying the mixed solution of the vegetable protein and the soapbark extract to obtain a vegetable protein-soapbark extract compound.
2. The method of claim 1, wherein the quillaja bark extract is used to improve the emulsifying properties of the plant protein, and the method comprises the steps of: the mass ratio of the plant protein to the natural quillaja bark extract in the step S1 is 100: 0.1-50.
3. The method for preparing a complex for improving the emulsifying properties of a plant protein according to claim 1, wherein: the vegetable protein comprises soybean protein, peanut protein, buckwheat protein, wheat protein, pea protein, cottonseed protein, rapeseed protein or potato protein.
4. The method for preparing a complex for improving the emulsifying properties of a plant protein according to claim 2, wherein: the soybean protein is soybean protein isolate or soybean protein concentrate.
5. The method for preparing a complex for improving the emulsifying properties of a plant protein according to claim 1, wherein: the quillaja bark extract is a liquid extract or a dehydrated extract solid.
6. The method for preparing a complex for improving the emulsifying properties of a plant protein according to claim 2, wherein: the drying method in the step S2 includes freeze drying, drying under normal pressure, drying under reduced pressure, spray drying or boiling drying.
7. A method for improving plant protein emulsifying property by cortex Gleditsiae Abnormalis extract is used in food system requiring protein emulsifying property.
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CN115404009A (en) * | 2022-10-11 | 2022-11-29 | 南京林业大学 | Soybean meal adhesive for plywood and preparation method thereof |
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CN115025230A (en) * | 2022-05-10 | 2022-09-09 | 仙乐健康科技股份有限公司 | Emulsifier composition and application thereof in emulsion |
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CN115404009A (en) * | 2022-10-11 | 2022-11-29 | 南京林业大学 | Soybean meal adhesive for plywood and preparation method thereof |
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