CN114304646A - Iron-protein nano-composite and preparation method and application thereof - Google Patents
Iron-protein nano-composite and preparation method and application thereof Download PDFInfo
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- CN114304646A CN114304646A CN202111630797.XA CN202111630797A CN114304646A CN 114304646 A CN114304646 A CN 114304646A CN 202111630797 A CN202111630797 A CN 202111630797A CN 114304646 A CN114304646 A CN 114304646A
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Abstract
The invention discloses an iron-protein nano-composite and a preparation method and application thereof. The preparation method comprises the following steps: uniformly mixing protein and water according to a proper mass-volume ratio, adjusting the pH value of the dispersion liquid, heating at 65-100 ℃ to obtain a modified protein dispersion liquid, adjusting the modified protein dispersion liquid to be neutral or alkaline, cooling, adding an iron/ferrous solution, stirring, standing, centrifuging, taking supernatant, and freeze-drying or spray-drying to obtain the iron-protein nano-composite with the average particle size of 50-150 nm. The method disclosed by the invention is simple in process, green and safe; and the prepared iron-protein nano-composite has stable property and good solubility, and has good development and application prospects.
Description
Technical Field
The invention belongs to the field of functional nano biological products, and particularly relates to an iron-protein nano composite as well as a preparation method and application thereof.
Background
Iron is an indispensable trace element of human body, is an important component of hemoglobin and various enzymes in the body, and participates in the important physiological metabolic processes of the human body such as the combination and transportation of oxygen, the maintenance of hematopoietic immune function and the like. Iron deficiency of human body is easy to cause iron deficiency anemia, which is a global public health problem. Iron deficiency anemia can lead to the impairment of labor and working ability of adults, retarded development of adolescent motor and cognitive abilities, increased risk of abortion in pregnant women, and the like.
The most common cause of iron deficiency is insufficient daily iron intake, and thus increasing iron intake in humans by iron supplements is the most common and reliable method for improving iron deficiency. However, the traditional micromolecular iron supplement agents such as ferrous sulfate, ferrous fumarate and other iron salts are active in property, have stimulation effect on intestines and stomach, are easy to generate side effects such as vomit, diarrhea and the like, and are not beneficial to long-term taking; in addition, the traditional iron salt is easy to hydrolyze and precipitate under the neutral pH value of the intestinal tract, and is easy to react with polyphenol or phytic acid and other substances in food to generate insoluble substances which cannot be absorbed by human bodies, so that the absorption and utilization rate is unstable. Therefore, the preparation method of the iron supplement agent with stable property and good solubility has good application prospect. Yanping of 2021 discloses a preparation method of a soybean protein fiber ferrous complex with pH response slow release and the complex, which is prepared by a soybean protein fiber solution, a ferrous solution and water-soluble ascorbic acid, and the preparation method specifically comprises the following steps: firstly, carrying out heating treatment on a soybean protein solution for 20-48h under the condition that the pH value is 1.5-2.4 to obtain a soybean protein fiber solution; the obtained soybean protein fiber solution, ferrous solution and water-soluble ascorbic acid are used for successfully preparing the compound with stable property and pH response slow-release effect. However, the heating treatment process for more than 20 hours is not favorable for the popularization and application of the method in industrial production.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention aims to provide a preparation method and application of an iron-protein nano-composite.
The primary object of the present invention is to provide an iron-protein nanocomposite.
Another object of the present invention is to provide a method for preparing the above-described iron-protein nanocomposite.
It is still another object of the present invention to provide the use of the above-described ferritin nanocomplexes.
The purpose of the invention is realized by at least one of the following technical solutions.
The invention provides a preparation method of an iron-protein nano-composite, which comprises the following steps:
(1) adding protein into water, and uniformly stirring to obtain a protein dispersion liquid;
(2) adjusting the pH value of the protein dispersion liquid in the step (1) to be alkaline, and heating the protein dispersion liquid in a stirring state to obtain a denatured protein dispersion liquid;
(3) adjusting the denatured protein dispersion liquid in the step (2) to be neutral or alkaline, and adding Fe2+Solution or Fe3+And (3) stirring the solution, standing, centrifuging to obtain a supernatant, and drying the supernatant to obtain the iron-protein nano-composite.
Further, the protein in the step (1) is one of rice protein, pea protein, whey protein, potato protein, mung bean protein, soybean protein, brown rice protein and the like.
Further, in the step (1), the mass-to-volume ratio of the protein to the water is (1-10):100 g/mL.
Further, in the step (1), the mass-to-volume ratio of the protein to the water is (2-6):100 (g/mL).
Further, in the step (1), the mass-to-volume ratio of the protein to the water is 4:100 (g/mL).
Further, in the step (2), the pH of the protein dispersion liquid is adjusted to be alkaline, so that the pH of the protein dispersion liquid after adjustment is 10-13.
Further, in the step (2), the temperature of the heating treatment is 65-100 ℃.
Further, in the step (2), the time of the heat treatment is 0.5h-1 h.
Further, in the step (2), the temperature of the heating treatment is 75-85 ℃.
Further, in the step (3), the denatured protein dispersion liquid in the step (2) is adjusted to be neutral or alkaline, so that the pH of the denatured protein dispersion liquid is 7-13.
Further, in the step (3), the denatured protein dispersion liquid in the step (2) is adjusted to be neutral or alkaline, so that the pH value of the denatured protein dispersion liquid is 7-12.
Further, in the step (3), the denatured protein dispersion liquid in the step (2) is adjusted to be neutral or alkaline, so that the pH value of the denatured protein dispersion liquid is 7-11.
Further, in the step (3), the Fe2+The solution is one of ferrous sulfate solution, ferrous fumarate solution, ferrous succinate solution, ferrous lactate solution, and ferrous chloride solution.
Further, in the step (3), the Fe2+The solution is ferrous sulfate solution.
Further, in the step (3), the Fe3+The solution is one of ferric chloride solution and ferric citrate solution.
Further, in the step (3), the Fe3+The solution is ferric chloride solution.
Further, said Fe in step (3)2+Solution or Fe3+Fe in solution2+Or Fe3+The mass ratio of the protein to the protein in the step (1) is (1-3): 100.
further, in the step (3), the standing treatment time is 2-10 h.
Further, in the step (3), the rotation speed of the centrifugal treatment is 3000-8000 g.
Further, in the step (3), the time of the centrifugal treatment is 15-30 min.
Further, in the step (3), the temperature of the centrifugal treatment is 15-25 ℃.
Further, in the step (3), the standing treatment time is 4-6 h.
Further, in the step (3), the rotation speed of the centrifugal treatment is 3000-5000 g.
Further, in the step (3), the time of the centrifugal treatment is 15-25 min.
Further, in the step (3), the drying manner includes freeze drying or spray drying.
Further, the temperature of the freeze drying is-50 to-40 ℃.
Further, the air inlet temperature of the spray drying is 160-180 ℃, and the air outlet temperature of the spray drying is 70-90 ℃.
Further, the average particle size of the iron-protein nano-complex in the step (3) is 50-150 nm.
The invention provides an iron-protein nano-composite prepared by the preparation method.
The invention provides the iron-protein nano-composite which can be applied to the preparation of foods or medicines for supplementing iron.
The iron-protein nano compound prepared by the invention can ensure that iron is stably and uniformly dispersed in neutral and alkaline systems, not only can be directly used for researching and developing functional food, but also can be used as a novel functional food ingredient with high nutritive value and physiological activity to be added into a food system.
Compared with the prior art, the invention has the following advantages and beneficial effects:
(1) the preparation method provided by the invention can well improve the solubility and stability of iron under neutral pH by combining protein and iron to form a uniform compound with smaller particle size, and has good application prospect.
(2) The preparation method provided by the invention has the advantages of simple process, greenness, safety, low energy consumption and good industrial prospect.
Drawings
FIG. 1 shows the separation of unheated soy protein and FeSO obtained in comparative example 14Solution reaction of the resulting product with that obtained in example 1Graph of particle size analysis results of the iron-soy protein nanocomposite.
FIG. 2 is a graph showing the results of particle size analysis of the iron-protein nanocomposite obtained in example 2-4.
Detailed Description
The following description of the embodiments of the present invention is provided in connection with the accompanying drawings and examples, but the invention is not limited thereto. It is noted that the processes described below, if not specifically described in detail, are all realizable or understandable by those skilled in the art with reference to the prior art. The reagents or apparatus used are not indicated to the manufacturer, and are considered to be conventional products available by commercial purchase.
Comparative example 1
(1) Adding Soy Protein Isolate (SPI) into water, wherein the mass-volume ratio of the protein to the water is 2:100g/mL, and uniformly stirring to obtain a soy protein isolate dispersion;
(2) adjusting pH of the soy protein isolate dispersion of step (1) to 11 with 2M NaOH, and adding 10mM FeSO4Solution to a final protein mass volume percent concentration of 1% (w/v, g/mL), final FeSO4Stirring for 10min at a concentration of 5mM, standing for 4h, centrifuging at 8000g and 20 deg.C for 15min, collecting supernatant, and freeze drying at-40 deg.C to obtain unheated SPI and FeSO4The solution reacted the resulting product, denoted as unheated SPI.
Example 1
The preparation method of the iron-protein nano-composite comprises the following steps:
(1) adding the soybean protein isolate into water, wherein the mass volume ratio of the protein to the water is 2:100g/mL, and uniformly stirring to obtain a soybean protein dispersion;
(2) and (2) regulating the pH value of the soy protein isolate dispersion liquid obtained in the step (1) to be 12 by using 2M NaOH, standing for 30min, and heating for 30min under a stirring state at the heating temperature of 85 ℃ to obtain a modified soy protein dispersion liquid.
(3) After cooling to room temperature, the pH of the modified soy protein isolate dispersion of step (2) was adjusted to 11 using 1M HCl, and 10mM FeSO was added4Solution, final protein mass volume hundredThe ratio concentration is 1% (w/v, g/mL), and the final FeSO4Stirring for 10min, standing for 4h, centrifuging at 8000g and 20 deg.C for 15min, and freeze drying the supernatant at-40 deg.C to obtain the iron-protein nanocomposite, and recording as heating SPI.
Comparative example 2
(1) Adding rice protein into water, wherein the mass volume ratio of the protein to the water is 4:100g/mL, and uniformly stirring to obtain a rice protein dispersion liquid;
(2) adjusting the pH value of the soybean protein dispersion liquid in the step (1) to 7 by using 2M NaOH, and adding 5mM FeCl2Solution to a final protein mass volume percent concentration of 2% (w/v, g/mL), final FeCl2Stirring at concentration of 2.5mM for 10min, standing for 6h, centrifuging at 3000g and 25 deg.C for 30min, and freeze drying the supernatant at-40 deg.C.
Example 2
The preparation method of the iron-protein nano-composite comprises the following steps:
(1) adding rice protein into water, wherein the mass volume ratio of the protein to the water is 4:100g/mL, and uniformly stirring to obtain a rice protein dispersion liquid;
(2) and (2) adjusting the pH value of the rice protein dispersion liquid obtained in the step (1) to 10 by using 2M NaOH, standing for 30min, and heating for 45min under a stirring state at the heating temperature of 100 ℃ to obtain a modified rice protein dispersion liquid.
(3) After cooling to room temperature, the pH of the denatured rice protein dispersion liquid of step (2) was adjusted to 7 with 1M HCl, and 5mM FeCl was added2Solution to a final protein mass volume percent concentration of 2% (w/v, g/mL), final FeCl2Stirring for 10min, standing for 6h, centrifuging at 3000g and 25 deg.C for 30min, collecting supernatant, and freeze drying at-40 deg.C to obtain Fe-protein nanocomposite (Fe-rice protein).
Comparative example 3
(1) Adding mung bean protein into water, wherein the mass volume ratio of the protein to the water is 10:100g/mL, and uniformly stirring to obtain a mung bean protein dispersion liquid;
(2) regulation of step (1) with 2M NaOHThe pH value of the mung bean protein dispersion liquid is 7.5, and 10mM FeCl is added2Solution to a final protein mass volume percent concentration of 5% (w/v, g/mL), final FeCl2Stirring for 10min at a concentration of 5mM, standing for 2h, centrifuging at 5000g and 15 deg.C for 20min, and freeze drying the supernatant at-40 deg.C.
Example 3
The preparation method of the iron-protein nano-composite comprises the following steps:
(1) adding mung bean protein into water, wherein the mass volume ratio of the protein to the water is 10:100g/mL, and uniformly stirring to obtain a soybean protein dispersion liquid;
(2) and (2) adjusting the pH value of the soybean protein dispersion liquid in the step (1) to 13 by using 2M NaOH, standing for 30min, and heating for 60min under a stirring state at the heating temperature of 65 ℃ to obtain a modified mung bean protein dispersion liquid.
(3) After cooling to room temperature, the pH of the denatured mung bean protein dispersion of step (2) was adjusted to 7.5 with 1M HCl, and 10mM FeCl was added2Solution to a final protein mass volume percent concentration of 5% (w/v, g/mL), final FeCl2Stirring for 10min, standing for 2h, centrifuging at 5000g and 15 deg.C for 20min, collecting supernatant, and freeze drying at-40 deg.C to obtain Fe-protein nanocomposite (Fe-mung bean protein).
Comparative example 4
(1) Adding pea protein into water, wherein the mass volume ratio of the pea protein to the water is 2:100g/mL, and uniformly stirring to obtain pea protein dispersion liquid;
(2) adjusting the pH value of the pea protein dispersion liquid in the step (1) to 7 by using 2M NaOH, and adding 5mM FeCl3Solution to a final protein mass volume percent concentration of 1% (w/v, g/mL), final FeCl3Stirring for 10min, standing for 10h, centrifuging at 8000g and 25 deg.C for 20min, and freeze drying the supernatant at-40 deg.C.
Example 4
The preparation method of the iron-protein nano-composite comprises the following steps:
(1) adding pea protein into water, wherein the mass volume ratio of the pea protein to the water is 2:100g/mL, and uniformly stirring to obtain pea protein dispersion liquid;
(2) and (2) regulating the pH value of the soybean protein dispersion liquid obtained in the step (1) to be 12 by using 2M NaOH, standing for 30min, and heating for 30min under a stirring state at the heating temperature of 85 ℃ to obtain a modified pea protein dispersion liquid.
(3) After cooling to room temperature, the pH of the modified soy protein dispersion of step (2) was adjusted to 7 using 1M HCl, and 5mM FeCl was added3Solution to a final protein mass volume percent concentration of 1% (w/v, g/mL), final FeCl3The concentration is 2.5mM, the mixture is stirred for 10min and then is kept stand for 10h, the mixture is centrifuged for 20min at 8000g and 25 ℃, and the supernatant is taken and freeze-dried at the temperature of minus 40 ℃ to obtain the iron-protein nano composite which is marked as Fe-pea protein.
The iron-protein nanocomposites prepared in comparative examples 1 to 4 and examples 1 to 4 were evaluated as follows, and the results are shown in table 1.
Determination of particle size and Polymer Dispersion Index (PDI): the average particle size and PDI of the obtained nanoparticles were determined using a Nano-particle size potentiostat (Malvern Nano-ZS).
As can be seen from fig. 1, the particle size of the iron-protein complex prepared from the heat-treated soy protein is unimodal and has narrow peak width, indicating that the complex has uniform size, which is favorable for stable dispersion; while the unheated soy protein was reacted with Fe under the same conditions2+The particle size distribution of the product obtained by the solution reaction is a short and wide peak, and the particle size distribution is wide and the uniformity is poor. As can be seen from FIG. 2, the rice protein, mung bean protein, pea protein and Fe after the heat treatment2+Solution or Fe3+The particle diameters of the compound formed by the solution reaction are all in monomodal distribution.
As is clear from Table 1, the average particle size of the iron-protein nanocomposite in examples 1 to 4 is 50 to 150nm, and the PDI is 0.20 to 0.30, which is preferable in terms of uniformity. Comparative examples 1-4 in which the protein and Fe were not heated2+Solution or Fe3+The average particle size of the product obtained by the solution reaction is larger than that of the corresponding embodiment, and the PDI is larger, which indicates that the system has poor uniformity and is not favorable for stable dispersion in water. The results show that the above process is carried outCan prepare the iron-protein nano-composite with small and uniform particle size and stable dispersion under neutral to alkaline conditions.
TABLE 1
The preparation method provided by the invention can well improve the solubility and stability of iron under neutral pH by combining protein and iron to form a uniform compound with smaller particle size, and has the advantages of simple process, greenness, safety, lower energy consumption and better industrial prospect. The above examples are only preferred embodiments of the present invention, which are intended to be illustrative and not limiting, and those skilled in the art should understand that they can make various changes, substitutions and alterations without departing from the spirit and scope of the invention.
Claims (10)
1. A method for preparing an iron-protein nanocomposite, comprising the steps of:
(1) adding protein into water, and uniformly stirring to obtain a protein dispersion liquid;
(2) adjusting the pH value of the protein dispersion liquid in the step (1) to be alkaline, and heating the protein dispersion liquid in a stirring state to obtain a denatured protein dispersion liquid;
(3) adjusting the denatured protein dispersion liquid in the step (2) to be neutral or alkaline, and adding Fe2+Solution or Fe3+And (3) stirring the solution, standing, centrifuging to obtain a supernatant, and drying the supernatant to obtain the iron-protein nano-composite.
2. The method for preparing an iron-protein nanocomposite as claimed in claim 1, wherein the protein in the step (1) is one of rice protein, pea protein, whey protein, potato protein, mung bean protein, soy protein and brown rice protein.
3. The method for preparing an iron-protein nanocomposite as claimed in claim 1, wherein the ratio of the mass of the protein to the volume of water in the step (1) is (1-10):100 g/mL.
4. The method for preparing the iron-protein nanocomposite as claimed in claim 1, wherein the pH of the protein dispersion is adjusted to be alkaline in the step (2) so that the pH of the protein dispersion after the adjustment is 10 to 13.
5. The method for preparing an iron-protein nanocomposite as claimed in claim 1, wherein the heat treatment temperature is 65 to 100 ℃ and the heat treatment time is 30 to 60min in the step (2).
6. The method for preparing the iron-protein nanocomposite as claimed in claim 1, wherein in the step (3), the denatured protein dispersion liquid obtained in the step (3) is adjusted to be neutral or alkaline so that the pH of the denatured protein dispersion liquid is 7 to 12.
7. The method for preparing the iron-protein nanocomposite as claimed in claim 1, wherein the Fe is added in the step (3)2+The solution is one of ferrous sulfate solution, ferrous fumarate solution, ferrous succinate solution, ferrous lactate solution, and ferrous chloride solution, and the Fe is3+The solution is one of ferric chloride solution and ferric citrate solution.
8. The method for preparing the iron-protein nanocomposite as claimed in claim 1, wherein in the step (3), the standing treatment time is 2-10h, the centrifugation rotation speed is 3000-8000g, the centrifugation time is 15-30min, and the centrifugation temperature is 15-25 ℃.
9. The iron-protein nanocomposite prepared by the preparation method according to any one of claims 1 to 8.
10. Use of the iron-protein nanocomposite according to claim 9 for preparing an iron-supplemented food or pharmaceutical product.
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Cited By (1)
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| CN115227716A (en) * | 2022-07-15 | 2022-10-25 | 南通励成生物工程有限公司 | Fibrous protein aggregate-iron compound and preparation method and application thereof |
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