CN114480544A - Preparation method of peanut peptide-calcium and zinc chelate - Google Patents
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Abstract
The invention belongs to the technical field of chemical synthesis of functional nutrient substances, and discloses a preparation method of peanut peptide-calcium and zinc chelates, which comprises the following steps of firstly preparing a peanut protein peptide solution, then carrying out chelation reaction, and finally separating and purifying, wherein the preparation process technology for preparing the peanut peptide-calcium and zinc chelates is established for the first time, the preparation method adopts specific process parameters, and the chelation rate of the finally obtained chelate can reach more than 50%; the operation is simple; the peanut peptide-calcium and zinc chelate is used as a novel bioactive peptide calcium nutrition enhancer, has good water solubility, is easy to be absorbed by human body, supplements calcium and zinc elements required by the human body, supplements nutrient components of the peanut peptide, and is a calcium and zinc simultaneous supplement product with good prospect.
Description
Technical Field
The invention belongs to the technical field of chemical synthesis of functional nutrients, and particularly relates to a preparation method of a peanut peptide-calcium and zinc chelate.
Background
The peanut peptide is a deep-processed product of peanut protein, has rich nutrient components, complete amino acid types and high essential amino acid content of 38.29 percent, and has a completely independent absorption mechanism. The peanut peptide has better solubility and stability than peanut protein, can be directly absorbed by small intestine, quickly supplements nutrition, directly participates in the synthesis of amino acid and protein of human body, promotes the active group of protein to play a role, enhances immunity, and has biological activity such as oxidation resistance. In addition, the peanut peptide has good metal ion chelating capacity, the preparation of peptide-calcium and zinc chelates by the peanut peptide has great advantages, and the absorption of calcium and zinc in the intestinal tract is hindered by the presence of dietary fiber and phytate in food, because the dietary fiber and phytate can form insoluble complexes with metal ions, and the bioavailability of the peanut peptide is reduced. The peanut peptide is chelated with calcium and zinc to form a chelate, so that the phenomena can be avoided, the solubility and stability of metal ions are obviously improved, and the absorption utilization rate of the metal ions is improved. Therefore, the calcium and zinc chelating peptide has high bioavailability and quick absorption, and is an ideal calcium and zinc co-supplement.
Calcium is a necessary nutrient element for the human body to maintain normal physiological metabolism and plays an extremely important role in the growth and development process of the human body. Calcium deficiency can lead to many diseases such as osteoporosis, rickets, etc. Because the dietary structure of residents in China has defects, people lack high-quality sources of calcium in the diet, and the calcium deficiency becomes a common nutritional disease in China. In fact, the daily average calcium intake of residents in China is only half of the recommended amount, and the daily average calcium intake is seriously insufficient. In view of the important role of calcium in the body, and the diseases caused by calcium deficiency, the preparation of calcium nutritional supplements has become a hot research topic.
Zinc is one of the essential trace elements for human body, has important regulation and control function on human health condition, and is praised as an intelligent element by medical and nutrition circles. The zinc deficiency can cause the problems of the human body such as the reduction of immunity, the growth retardation, the reduction of appetite, the dysfunction of the central nervous system and the like. The peptide chelated zinc formed by chelating zinc and peptide has a special monocyclic chelating structure, and compared with inorganic zinc salt, the zinc in the peptide zinc chelate is more easily absorbed by a human body, has stronger stability, and has good effects of supplementing zinc and improving the immunity of the organism.
At present, the research on peanuts is mainly used for oil extraction, and the protein content of peanut meal after oil extraction is very rich, but the peanut meal is thrown away as feed or in the form of waste, so that a large amount of resources are wasted, and the environment is polluted. If the peanut meal after oil extraction can be comprehensively utilized and the protein in the peanut meal is utilized to prepare the calcium and zinc chelating peptide, the problem of resource waste of the peanut meal can be solved, and a high-efficiency calcium and zinc ion supplement can be provided to meet the increasing needs of people.
Disclosure of Invention
In view of the problems raised by the background art described above, the present invention has an object to: aims at providing a preparation method of peanut peptide-calcium and zinc chelate. In order to achieve the technical purpose, the technical scheme adopted by the invention is as follows:
a preparation method of peanut peptide-calcium and zinc chelate comprises the following steps:
s1, preparing a peanut protein peptide solution: preparing a peanut protein peptide solution with the concentration of 10 g/L;
s11, preprocessing the peanut protein: preparing a peanut protein solution with the concentration of 50g/L by using deionized water, and carrying out ultrasonic treatment;
s12, enzymolysis: adding alkaline protease into the peanut protein solution obtained in the step S11, adding 3% of alkaline protease into every 1g of peanut protein dry weight by taking the peanut protein dry weight in the peanut protein solution as a reference, adjusting the pH value to 8, carrying out boiling water bath treatment at the temperature of 100-120 ℃ for 10 minutes, adjusting the pH value of an enzymolysis solution to 7.0 after 3 hours of enzymolysis, adding 3% of flavourzyme, carrying out enzymolysis at the temperature of 55 ℃ for 3 hours, and then inactivating the protease to obtain the enzymolysis solution; then cooling at room temperature, adjusting pH to 4.5 with hydrochloric acid solution after cooling, precipitating the peanut protein without enzymolysis by using isoelectric point, centrifuging the sample, taking supernatant, and freeze-drying to obtain peanut protein peptide;
s13, preparation: modifying the peanut protein peptide obtained in the S12 with sodium tripolyphosphate, and preparing a peanut protein peptide solution with the concentration of 10g/L by using deionized water;
s2, carrying out chelation reaction;
s21, adjusting the pH value: adjusting the pH value of the peanut protein peptide solution prepared in the step S1 to 5-7, and mixing;
s22, mixing: mixing the pH value of the mixed solution with anhydrous calcium chloride and zinc sulfate;
s23, water bath: carrying out chelation reaction for 50 minutes under the water bath condition of 50 ℃ to obtain a chelation reaction product solution;
s3, separating and purifying;
s31, filtering: filtering the chelating reaction product solution obtained in the step S2 by using a filtering membrane to obtain a filtrate;
s32, ethanol separation: mixing the filtrate with 10 times of edible ethanol, and centrifuging;
s33, obtaining a finished product: and (3) taking the precipitate obtained after centrifugation, redissolving in deionized water, and freeze-drying to obtain the peanut peptide-calcium and zinc chelate.
Further limiting, in step S11, the setting value of the ultrasonic processing device is 1500W power, 20min time, 4S ultrasonic processing, 2S intermittent processing, and 50 ℃ maximum temperature, so that the oscillating mixing effect is good.
Further limiting, in step S12, the conditions required for high-temperature inactivation of protease are that the protease is treated in a boiling water bath at 100 ℃ for 10 minutes, so that the quality of the enzymatic hydrolysate is ensured while the inactivation effect is ensured.
Further, in step S12, the centrifugation is performed for 15 minutes at 5000r/min, which is a good separation effect.
Further limiting, in step S22, the peanut protein hydrolysate is dissolved in deionized water to obtain a solution with a mass fraction of 5%, and the solution is mixed with calcium chloride and zinc sulfate, so as to ensure the solution ratio.
Further limiting, in step S21, the pH value of the mixed solution is in the range of 6, in such a way as to ensure the subsequent reaction.
Further limiting, in the step S23, the temperature ranges from 40 ℃ to 80 ℃, so that the reaction effect is ensured.
Further limiting, in the step S32, after mixing, standing for 1-1.5 h, and then centrifuging, so that the separation effect after precipitation is better.
The invention has the following beneficial effects:
the preparation method adopts specific process parameters, and the chelation rate of the finally obtained chelate can reach more than 50 percent; the operation is simple; the peanut peptide-calcium and zinc chelate is used as a novel bioactive peptide calcium nutrition enhancer, has good water solubility, is easy to be absorbed by human body, supplements calcium and zinc elements required by the human body, supplements nutrient components of the peanut peptide, and is a calcium and zinc simultaneous supplement product with good prospect.
Drawings
The invention is further illustrated by the non-limiting examples given in the accompanying drawings;
FIG. 1 is a schematic block diagram of an embodiment of a method for preparing peanut peptide-calcium and zinc chelates according to the present invention.
Detailed Description
In order that those skilled in the art can better understand the present invention, the following technical solutions are further described with reference to the accompanying drawings and examples.
As shown in fig. 1, the preparation method of the peanut peptide-calcium and zinc chelate of the present invention comprises the following steps:
s1, preparing a peanut protein peptide solution: preparing a peanut protein peptide solution with the concentration of 10 g/L;
s11, preprocessing the peanut protein: preparing a peanut protein solution with the concentration of 50g/L by using deionized water, and carrying out ultrasonic treatment;
s12, enzymolysis: adding alkaline protease into the peanut protein solution obtained in the step S11, adding 3% of alkaline protease into every 1g of peanut protein dry weight by taking the peanut protein dry weight in the peanut protein solution as a reference, adjusting the pH value to 8, carrying out boiling water bath treatment at the temperature of 100-120 ℃ for 10 minutes, adjusting the pH value of an enzymolysis solution to 7.0 after 3 hours of enzymolysis, adding 3% of flavourzyme, carrying out enzymolysis at the temperature of 55 ℃ for 3 hours, and then inactivating the protease to obtain the enzymolysis solution; then cooling at room temperature, adjusting pH to 4.5 with hydrochloric acid solution after cooling, precipitating the peanut protein without enzymolysis by using isoelectric point, centrifuging the sample, taking supernatant, and freeze-drying to obtain peanut protein peptide;
s13, preparation: modifying the peanut protein peptide obtained in the S12 with sodium tripolyphosphate, and preparing a peanut protein peptide solution with the concentration of 10g/L by using deionized water;
s2, carrying out chelation reaction;
s21, adjusting the pH value: adjusting the pH value of the peanut protein peptide solution prepared in the step S1 to 5-7, and mixing;
s22, mixing: mixing the pH value of the mixed solution with anhydrous calcium chloride and zinc sulfate;
s23, water bath: carrying out chelation reaction for 50 minutes under the water bath condition of 50 ℃ to obtain a chelation reaction product solution;
s3, separating and purifying;
s31, filtering: filtering the chelating reaction product solution obtained in the step S2 by using a filtering membrane to obtain a filtrate;
s32, ethanol separation: mixing the filtrate with 10 times of edible ethanol, and centrifuging;
s33, obtaining a finished product: and (3) taking the precipitate obtained after centrifugation, redissolving in deionized water, and freeze-drying to obtain the peanut peptide-calcium and zinc chelate.
In this embodiment, when a method for preparing peanut peptide-calcium and zinc chelate is used, ultrasonic-assisted enzymolysis and phosphorylation modification of peanut protein: adding deionized water into the peanut protein according to the ratio of 1:10, and performing ultrasonic pretreatment for 20 min; adjusting the pH value to be about 6.0 accurately, adding 3.0% of alkaline protein, performing enzymolysis at 55 ℃ for 3h, adjusting the pH value to 7.0, adding 3% of flavourzyme, performing enzymolysis at 55 ℃ for 3h, inactivating enzyme in a boiling water bath for 10 min, cooling at room temperature, adjusting the pH value to be 4.5 accurately after cooling, precipitating peanut protein which is not subjected to enzymolysis by using an isoelectric point, centrifuging a sample for 15min under the condition of 5000r/min, taking supernatant, preparing a peanut protein peptide solution, and performing Sodium Tripolyphosphate (STP) modification on the peanut protein peptide subjected to enzymolysis to enable some amino acid residue groups to be substituted by phosphate groups to obtain final peanut protein peptide;
dissolving peanut protein zymolyte into a solution with the mass fraction of 5% by using deionized water, mixing the solution with calcium chloride and zinc sulfate, adjusting the pH value of the mixed solution to 4.0-7.0, and carrying out chelation for 50 minutes under the water bath condition of the temperature of 40-80 ℃ to obtain a solution of a chelation reaction product; after the reaction is finished, centrifugally precipitating to obtain supernatant, then adding ethanol into the supernatant, standing for one hour, centrifugally precipitating, and drying the precipitate to obtain calcium and zinc chelating peptide;
the mass ratio of the peanut protein peptide to the calcium to the zinc in the peanut protein peptide solution is 1:2, 1:1, 2:1, 5:1 and 10: 1;
wherein, the determination of the calcium ion content:
the specific detection method is as shown in the EDTA method in GB5009.92-2016 (determination of calcium in food).
Chelate ratio (%)
= (calcium content/total calcium content in peptide-calcium, zinc chelate)
100%
=（CV2/CV1）
100%
=（V2/V1）
100%
Wherein, C: standard EDTA solution concentration, mol/L;
v2: titrating the volume of EDTA solution consumed by chelating calcium, mL;
v1: volume of EDTA solution consumed for titrating the total amount of calcium, mL
Wherein, the determination of the total calcium content:
0.1mL of the reaction solution was pipetted into a conical flask, 20mL of distilled water was added, 2mL of triethanolamine, KOH1mL, and 1mL of calcium indicator were added in sequence, the mixture was shaken gently and immediately titrated with EDTA, the titration end point was the change of the solution from magenta to pure blue, and the volume of EDTA consumed was recorded as V1.
Wherein, the determination of the calcium content in the chelate:
taking 1mL of reaction solution, adding 10 times of volume of absolute ethyl alcohol, centrifuging for 15min under the condition of the rotating speed of 5000r/min, and removing supernatant. To the precipitate was added 10mL of distilled water and shaken up with a vortex shaker. Sucking 1mL of liquid into a conical flask, adding 20mL of distilled water, sequentially adding 2mL of triethanolamine, 2mL of KOH1mL and 1mL of calcium indicator, shaking up gently, immediately titrating with EDTA (ethylene diamine tetraacetic acid), wherein the titration endpoint is that the solution changes from purple red to pure blue, and recording the volume of the EDTA consumed and recording the volume as V2.
Determination of the chelating rate of zinc ions:
EDTA complexation titration method is adopted. Weighing a certain amount of chelate powder, placing the chelate powder in a beaker, adding 50mL of water and 3mL of 6 mol of hydrochloric acid per liter, carrying out ultrasonic treatment until the chelate powder is completely dissolved, cooling to a constant volume of 100, taking 10mL of the chelate powder in a triangular flask, adding 10mL of H3-NH4Cl (pH =10) buffer solution and 2 drops of chrome black T indicator, titrating the mixture by using EDTA solution until the color of the mixture is changed from wine red to blue, recording the number of milliliters consumed by titration, carrying out parallel measurement on each sample solution for 3 times, taking an average value, and calculating the chelating rate of zinc ions.
In the formula, M1 is the mass of zinc ions in peptide chelated calcium and zinc;
c is the concentration of EDTA standard solution;
v0 is the volume mL of EDTA consumed in the blank test;
v1 is the volume mL of EDTA consumed on average during the measurement.
Preferably, in step S11, the setting value of the ultrasonic processing device is 1500W power, 20min time, 4S ultrasonic processing, 2S intermittent processing, and 50 ℃ maximum temperature, so that the oscillating mixing effect is good, and actually, the setting value of the ultrasonic processing device can be considered according to specific situations.
In step S12, the conditions required for inactivating the protease at high temperature are preferably boiling water bath treatment at 100 ℃ for 10 minutes, so as to ensure the quality of the enzymolysis solution while ensuring the inactivation effect, and in fact, the temperature and time can be considered according to specific conditions.
In step S12, the centrifugation is preferably performed at 5000r/min for 15 minutes, which is preferable because the separation effect is good, and the centrifugation conditions can be considered according to the circumstances.
Preferably, in step S22, the peanut proteolysis product is dissolved by deionized water to obtain a solution with a mass fraction of 5%, and the solution is mixed with calcium chloride and zinc sulfate in such a way as to ensure the solution ratio, and in such a way, the solution ratio is ensured, and in fact, the mass fraction of the proteolysis product can also be considered according to specific conditions.
Preferably, in step S21, the pH value of the mixed solution is in the range of 6, in such a way as to ensure the subsequent reaction, and in fact, the pH value of the mixed solution may be considered according to specific situations.
Preferably, in step S23, the temperature is in the range of 40 ℃ to 80 ℃, in this way, the reaction effect is ensured, and in fact, the temperature value can be considered according to specific situations.
Preferably, in step S32, the mixture is left to stand for 1 to 1.5 hours and then centrifuged, so that the separation effect after precipitation is better, and in fact, the standing time can be considered according to specific conditions.
The foregoing embodiments are merely illustrative of the principles of the present invention and its efficacy, and are not to be construed as limiting the invention. Any person skilled in the art can modify or change the above-mentioned embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which can be made by those skilled in the art without departing from the spirit and technical spirit of the present invention be covered by the claims of the present invention.
Claims (8)
1. A preparation method of peanut peptide-calcium and zinc chelate is characterized by comprising the following steps:
s1, preparing a peanut protein peptide solution: preparing a peanut protein peptide solution with the concentration of 10 g/L;
s11, preprocessing the peanut protein: preparing a peanut protein solution with the concentration of 50g/L by using deionized water, and carrying out ultrasonic treatment;
s12, enzymolysis: adding alkaline protease into the peanut protein solution obtained in the step S11, adding 3% of alkaline protease into every 1g of peanut protein dry weight by taking the peanut protein dry weight in the peanut protein solution as a reference, adjusting the pH value to 8, carrying out boiling water bath treatment at the temperature of 100-120 ℃ for 10 minutes, adjusting the pH value of an enzymolysis solution to 7.0 after 3 hours of enzymolysis, adding 3% of flavourzyme, carrying out enzymolysis at the temperature of 55 ℃ for 3 hours, and then inactivating the protease to obtain the enzymolysis solution; then cooling at room temperature, adjusting pH to 4.5 with hydrochloric acid solution after cooling, precipitating non-enzymatic peanut protein by isoelectric point, centrifuging the sample, collecting supernatant, and freeze drying to obtain peanut protein peptide;
s13, preparation: modifying the peanut protein peptide obtained in the S12 with sodium tripolyphosphate, and preparing a peanut protein peptide solution with the concentration of 10g/L by using deionized water;
s2, carrying out chelation reaction;
s21, adjusting the pH value: adjusting the pH value of the peanut protein peptide solution prepared in the step S1 to 5-7, and mixing;
s22, mixing: mixing the pH value of the mixed solution with anhydrous calcium chloride and zinc sulfate;
s23, water bath: carrying out chelation reaction for 50 minutes under the water bath condition of 50 ℃ to obtain a chelation reaction product solution;
s3, separating and purifying;
s31, filtering: filtering the chelating reaction product solution obtained in the step S2 by using a filtering membrane to obtain a filtrate;
s32, ethanol separation: mixing the filtrate with 10 times of edible ethanol, and centrifuging;
s33, obtaining a finished product: and (3) taking the precipitate obtained after centrifugation, redissolving in deionized water, and freeze-drying to obtain the peanut peptide-calcium and zinc chelate.
2. The method for preparing peanut peptide-calcium-zinc chelate according to claim 1, wherein the method comprises the following steps: in step S11, the setting values of the ultrasonic processing apparatus are power 1500W, time 20min, ultrasonic 4S, pause 2S, and maximum temperature 50 ℃.
3. The method for preparing peanut peptide-calcium-zinc chelate according to claim 2, wherein the method comprises the following steps: in step S12, the conditions required for high temperature inactivation of protease are boiling water bath treatment at 100 ℃ for 10 minutes.
4. The method for preparing peanut peptide-calcium-zinc chelate according to claim 3, wherein the method comprises the following steps: in step S12, the centrifugation is performed for 15 minutes at 5000 r/min.
5. The method for preparing peanut peptide-calcium-zinc chelate according to claim 4, wherein the method comprises the following steps: in step S22, the peanut protein hydrolysate is dissolved in deionized water to obtain a solution with a mass fraction of 5%, and the solution is mixed with calcium chloride and zinc sulfate.
6. The method for preparing peanut peptide-calcium-zinc chelate according to claim 5, wherein the method comprises the following steps: in step S21, the pH range of the mixed solution is 6.
7. The method for preparing peanut peptide-calcium-zinc chelate according to claim 6, wherein the method comprises the following steps: in step S23, the temperature range is 40-80 ℃.
8. The method for preparing peanut peptide-calcium-zinc chelate according to claim 7, wherein the method comprises the following steps: and step S32, mixing, standing for 1-1.5 h, and centrifuging.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115316672A (en) * | 2022-09-23 | 2022-11-11 | 吉林农业大学 | Preparation method of peanut peptide-calcium chelate |
| CN115501132A (en) * | 2022-06-28 | 2022-12-23 | 广州大学 | Preparation method of auricularia polytricha polysaccharide-zinc compound |
| CN115820776A (en) * | 2022-12-12 | 2023-03-21 | 青岛农业大学 | Peanut peptide calcium chelate as well as preparation method and application thereof |
| CN117099869A (en) * | 2023-06-27 | 2023-11-24 | 河南工业大学 | Preparation method of wheat protein peptide chelated calcium |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115501132A (en) * | 2022-06-28 | 2022-12-23 | 广州大学 | Preparation method of auricularia polytricha polysaccharide-zinc compound |
| CN115316672A (en) * | 2022-09-23 | 2022-11-11 | 吉林农业大学 | Preparation method of peanut peptide-calcium chelate |
| CN115316672B (en) * | 2022-09-23 | 2023-11-07 | 吉林农业大学 | Preparation method of peanut peptide-calcium chelate |
| CN115820776A (en) * | 2022-12-12 | 2023-03-21 | 青岛农业大学 | Peanut peptide calcium chelate as well as preparation method and application thereof |
| CN117099869A (en) * | 2023-06-27 | 2023-11-24 | 河南工业大学 | Preparation method of wheat protein peptide chelated calcium |
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