CN109880873B - Method for improving emulsifying activity of perilla seed protein - Google Patents
Method for improving emulsifying activity of perilla seed protein Download PDFInfo
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Abstract
The invention discloses a method for improving the emulsifying activity of perilla seed protein, belonging to the technical field of perilla seed protein modification. The method comprises the following steps: 1) adjusting the moisture of the perilla seed meal, and performing ultrahigh pressure pretreatment after adjusting the moisture; 2) crushing and sieving the perilla seed meal subjected to ultrahigh pressure pretreatment, adding water, mixing to prepare a mixed solution with the concentration of 4-20%, adjusting the pH and the temperature, adding protease for enzymolysis, cooling to room temperature after enzymolysis, and adjusting the pH value to be neutral; 3) and carrying out ultrafiltration treatment on the enzymolysis liquid to obtain perilla seed protein with the molecular weight cutoff of 5-30 kDa, and concentrating and spray drying the perilla seed protein to obtain the perilla seed protein with high emulsifying activity. The method has the characteristics of simple production process, less enzyme consumption, short enzymolysis time and low production cost.
Description
Technical Field
The invention belongs to the technical field of perilla seed protein modification, and particularly relates to a method for improving the emulsifying activity of perilla seed protein.
Background
Emulsifying activity refers to the ability of a protein product to bind oil and water together to form an emulsion. Three general evaluation criteria are used, namely the Emulsification Activity Index (EAI), the Emulsification Capacity (EC) and the emulsification stability. In a food emulsification system, protein can reduce the tension of an oil-water interface, thereby preventing the aggregation of oil droplets in the system. The study finds that the perillartine protein isolate globulin and albumin contain a large amount of hydrophilic groups and hydrophobic groups, and the special structures determine the surface activity of the perillartine protein and are easy to form stable emulsion.
The ultrahigh pressure technology is a pure physical process in the high pressure treatment process, has the characteristics of instantaneous compression, uniform action, safe operation, low energy consumption and the like, and is widely applied to industrial production. Meanwhile, ultrahigh pressure is also applied to the physical modification of proteins. In the high-pressure treatment process, the volume of the material in a liquid medium is compressed, and the high static pressure generated by the ultrahigh pressure can change the non-covalent bonds such as hydrogen bonds, ionic bonds, hydrophobic bonds and the like of the formed biopolymer three-dimensional structure. The protein folds, linearizes and re-crosslinks a high-order structure under the action of the protein, and the spatial conformation of the protein is changed, so that the emulsibility of the protein is influenced.
Domestic scholars such as Jiangweinxin research the emulsifying activity and the emulsifying stability of perilla seed meal under different enzymolysis conditions, and the results show that the emulsifying activity and the emulsifying stability index of perilla seed protein are improved at the beginning of hydrolysis, but small peptides formed by hydrolysis are not easy to be adsorbed on an oil-water interface along with the increase of the hydrolysis degree, so that the emulsifying activity and the emulsifying stability are reduced. The limited hydrolyzed perilla seed protein with a degree of hydrolysis of 3% has the best emulsifying activity when treated with alkaline protease, and the emulsifying activity is improved by more than 40% compared with that of perilla seed protein in a measured pH range. The microwave-assisted enzyme hydrolysis method is adopted, and when the hydrolysis degree is 15%, the emulsifying capacity index and the emulsifying stability index are the largest. The royal winner combines 1% of xanthan gum and 1% of guar gum with perilla seed meal respectively, and the emulsifying activity performance of perilla seed protein before and after combination is determined, and the result shows that the emulsifying activity index and the emulsifying stability of the perilla seed protein can be improved by both the royal winner and the perilla seed meal.
The existing perilla seed protein modification method has the problems of low modification degree, large enzyme dosage, long enzymolysis time and the like, and is not suitable for industrial continuous production. Therefore, it is important to find a method which is fast, convenient and suitable for industrial continuous production. At present, researches on improving the emulsifying activity of perilla seed protein by combining ultrahigh pressure pretreatment with enzyme method and modification are not reported.
Disclosure of Invention
The invention aims to provide a method for improving the emulsifying activity of perilla seed protein, which can achieve the aims of improving the emulsifying activity of the perilla seed protein, simplifying the process and reducing the enzyme dosage and the enzymolysis time.
The invention is realized by the following technical scheme:
a method for improving emulsifying activity of perilla seed protein comprises the following steps:
1) adjusting the moisture of the perilla seed meal, and performing ultrahigh pressure pretreatment after adjusting the moisture;
2) pulverizing the perilla seed meal pretreated under ultrahigh pressure, sieving, adding water, mixing to obtain a perilla seed meal mixed solution, adjusting pH and temperature, adding protease for enzymolysis, cooling to room temperature after enzymolysis, and adjusting pH to neutral;
3) and carrying out ultrafiltration treatment on the enzymolysis liquid to obtain perilla seed protein with the molecular weight cutoff of 5-30 kDa, and concentrating and spray drying the perilla seed protein with the molecular weight cutoff of 5-30 kDa to prepare the perilla seed protein with high emulsifying activity.
Preferably, in the step 1), the water content of the perilla seed meal is adjusted to 4-20%.
Preferably, in the step 1), the pressure used for the ultrahigh pressure pretreatment is 100-600 MPa, and the treatment time is 15-60 min.
Preferably, in the step 2), the sieving is to sieve the crushed perilla seed meal by a sieve of 40-200 meshes.
Preferably, in step 2), the protease is one or more of trypsin, pepsin, alkaline protease and papain.
Preferably, in the step 2), the pH value of the perilla seed meal mixed solution is adjusted to 2-11, and the temperature is adjusted to 35-65 ℃.
Preferably, in the step 2), the enzyme adding amount of enzymolysis is 1-50 mg/g of substrate content, and the enzymolysis time is 30-240 min.
Preferably, in the step 3), the temperature of an air inlet of drying equipment for spray drying is set to be 150-220 ℃.
Compared with the prior art, the invention has the following beneficial technical effects:
the method adopts ultrahigh pressure pretreatment combined with enzyme method to prepare the perilla seed protein with high emulsifying activity, perilla seed meal is subjected to high pressure action to partially degrade the protein or break a disulfide bond, the structure of the perilla seed protein is damaged to a certain degree, the change of the existence state of the disulfide bond and the aggregation and depolymerization phenomena of protein particles occur in the process, secondary bonds of the perilla seed meal are opened, protein molecules are further stretched to expose enzymolysis sites, the subsequent enzymolysis is facilitated, the protein peptide bond is broken due to enzymolysis, the molecular weight of the protein is reduced, charged groups are increased, and hydrophobic groups in the protein are exposed due to the change of the molecular structure, so that the emulsifying activity of the perilla seed protein is improved. The ultrahigh pressure treatment not only absorbs energy by protein, but also has a huge energy release process when the pressure is reduced, so that the increase of the conformational flexibility of the enzyme is promoted, and although the change of the bond energy and the bond angle of the protein is tiny, the integral structure of the protein cannot be collapsed, if the change of the bond occurs at the active site of the enzyme, the enzyme activity will be lost according to the key locking principle. If the bond change does not occur at the active site, the pressurization process can increase the exposed active site of the enzyme, and thus the enzymatic efficiency of the enzyme is stronger. The method has the characteristics of simple production process, less enzyme consumption, short enzymolysis time and low production cost.
Further, the pressure used for ultrahigh pressure pretreatment is 100-600 MPa, the pressure is an important factor influencing the ultrahigh pressure treatment effect, the pressure rises sharply, partial degradation or disulfide bond breakage of protein is caused, protein molecules are further stretched, enzymolysis sites are exposed, and subsequent enzymolysis is facilitated. The pressure is too low, the degradation degree inside protein molecules is not enough, and the subsequent enzymolysis is not facilitated; when the pressure is too high, the protein denaturation degree is too high, and a certain emulsification effect cannot be achieved.
Further, the enzymolysis time is set to be 30-240 min, and when the enzymolysis time is shorter, the modification effect of the perilla seed protein is poorer; however, the time is too long, and the modification effect is not ideal, so that the interval range selected by the invention can achieve good effect.
Detailed Description
The present invention will now be described in further detail with reference to specific examples, which are intended to be illustrative, but not limiting, of the invention.
Example 1
A method for improving emulsifying activity of perilla seed protein, which comprises the following steps:
(1) adjusting the moisture of the perilla seed meal to 4%, and then performing ultrahigh pressure pretreatment, wherein the pressure used for the ultrahigh pressure pretreatment is 600MPa, and the treatment time is 15 min;
(2) crushing the perilla seed meal pretreated under the ultrahigh pressure in the step (1), sieving the crushed perilla seed meal with a 40-mesh sieve, adding water, mixing to prepare a 6% perilla seed meal solution, adjusting the pH to 2, adjusting the temperature to 35 ℃, adding pepsin with the substrate concentration of 1mg/g for enzymolysis for 240min, cooling to room temperature after enzymolysis, and adjusting the pH to be neutral;
(3) performing ultrafiltration treatment on the enzymolysis liquid to obtain perilla seed protein with the molecular weight cutoff of 20kDa, and concentrating and spray drying the trapped protein (the air inlet temperature is 150 ℃) to obtain the perilla seed protein with high emulsifying activity.
The Emulsifying Activity Index (EAI) of unmodified perillartine is 13.28m at pH 7, compared with modified perillartine2The/g is increased to 43.64m2/g。
Example 2
A method for increasing the emulsifying activity of perillartine protein, which comprises the following steps:
(1) adjusting the moisture of the perilla seed meal to 20%, and then performing ultrahigh pressure pretreatment, wherein the pressure used for the ultrahigh pressure pretreatment is 300MPa, and the treatment time is 35 min;
(2) crushing the perilla seed meal pretreated under the ultrahigh pressure in the step (1), sieving the crushed perilla seed meal with a 70-mesh sieve, adding water, mixing to prepare a 10% perilla seed meal solution, adjusting the pH to 7, adjusting the temperature to 40 ℃, adding trypsin with the substrate concentration of 10mg/g for enzymolysis for 200min, cooling to room temperature after enzymolysis, and adjusting the pH to be neutral;
(3) performing ultrafiltration treatment on the enzymolysis liquid to obtain perilla seed protein with the molecular weight cutoff of 30kDa, and concentrating and spray drying the trapped protein (the air inlet temperature is 170 ℃) to obtain the perilla seed protein with high emulsifying activity.
The Emulsifying Activity Index (EAI) of unmodified perillartine is 11.34m at pH 7, compared with modified perillartine2The/g is increased to 47.37m2/g。
Example 3
A method for increasing the emulsifying activity of perillartine protein, which comprises the following steps:
(1) adjusting the water content of the perilla seed meal, adjusting the water content to 15%, and then performing ultrahigh pressure pretreatment, wherein the pressure used for the ultrahigh pressure pretreatment is 200MPa, and the treatment time is 40 min.
(2) And (2) crushing the perilla seed meal pretreated under the ultrahigh pressure in the step (1), sieving the crushed perilla seed meal with a 120-mesh sieve, adding water, mixing to prepare a 10% perilla seed meal solution, adjusting the pH to 11, adjusting the temperature to 45 ℃, adding alkaline protease with the substrate concentration of 30mg/g for enzymolysis for 160min, cooling to room temperature after the enzymolysis, and adjusting the pH to be neutral.
(3) Performing ultrafiltration treatment on the enzymolysis liquid to obtain perilla seed protein with the molecular weight cutoff of 20kDa, and concentrating and spray drying the trapped protein (the air inlet temperature is 220 ℃) to obtain the perilla seed protein with high emulsifying activity.
The Emulsifying Activity Index (EAI) of unmodified perillartine is 11.83m at pH 7, compared with modified perillartine2The/g is increased to 48.66m2/g。
Example 4
A method for improving emulsifying activity of perilla seed protein, which comprises the following steps:
(1) adjusting the moisture of the perilla seed meal to 20%, and then performing ultrahigh pressure pretreatment at 100MPa for 60 min;
(2) pulverizing the perilla seed meal pretreated under the ultrahigh pressure in the step (1), sieving with a 150-mesh sieve, adding water, mixing to prepare a 10% perilla seed meal solution, adjusting the pH to 8, adjusting the temperature to 55 ℃, adding a mixture of alkaline protease and papain with the substrate concentration of 40mg/g, performing enzymolysis for 80min, cooling to room temperature after enzymolysis, adjusting the pH to neutral,
(3) performing ultrafiltration treatment on the enzymolysis liquid to obtain perilla seed protein with the cut-off molecular weight of 10kDa, and concentrating and spray drying the cut-off protein (the air inlet temperature is 200 ℃) to obtain the perilla seed protein with high emulsifying activity.
The Emulsifying Activity Index (EAI) of unmodified perillartine is from 12.65m at pH 7, compared with modified perillartine2The/g is increased to 47.93m2/g。
Example 5
A method for improving emulsifying activity of perilla seed protein, which comprises the following steps:
(1) adjusting the water content of the perilla seed meal to 20%, and then performing ultrahigh pressure pretreatment, wherein the pressure for the ultrahigh pressure pretreatment is 300MPa, and the treatment time is 45 min;
(2) pulverizing the perilla seed meal pretreated under the ultrahigh pressure in the step (1), sieving with a 200-mesh sieve, adding water, mixing to prepare a 15% perilla seed meal solution, adjusting the pH to 4, adjusting the temperature to 65 ℃, adding pepsin with the substrate concentration of 50mg/g, performing enzymolysis for 30min, cooling to room temperature after enzymolysis, adjusting the pH to neutral,
(3) performing ultrafiltration treatment on the enzymolysis liquid to obtain perilla seed protein with the cut-off molecular weight of 10kDa, and concentrating and spray drying the cut-off protein (the air inlet temperature is 170 ℃) to obtain the perilla seed protein with high emulsifying activity.
The Emulsifying Activity Index (EAI) of unmodified perillartine is 11.35m at pH 7, compared with modified perillartine2The/g is increased to 46.48m2/g。
Claims (6)
1. A method for improving the emulsifying activity of perilla seed protein is characterized by comprising the following steps:
1) adjusting the moisture of the perilla seed meal, and performing ultrahigh pressure pretreatment after adjusting the moisture; the pressure for ultrahigh pressure pretreatment is 100-600 MPa, and the treatment time is 15-60 min;
2) pulverizing the perilla seed meal pretreated under ultrahigh pressure, sieving, adding water, mixing to obtain a perilla seed meal mixed solution, adjusting pH and temperature, adding protease for enzymolysis, cooling to room temperature after enzymolysis, and adjusting pH to neutral; the enzyme adding amount of enzymolysis is 1-50 mg/g of substrate content, and the enzymolysis time is 30-240 min;
3) and carrying out ultrafiltration treatment on the enzymolysis liquid to obtain perilla seed protein with the molecular weight cutoff of 5-30 kDa, and concentrating and spray drying the perilla seed protein with the molecular weight cutoff of 5-30 kDa to prepare the perilla seed protein with high emulsifying activity.
2. The method for improving the emulsifying activity of perilla seed protein according to claim 1, wherein in the step 1), the water content of perilla seed meal is adjusted to 4-20%.
3. The method for improving the emulsifying activity of perilla seed protein according to claim 1, wherein the sieving in the step 2) is to sieve the crushed perilla seed meal with a 40-200 mesh sieve.
4. The method for improving the emulsifying activity of perilla seed protein according to claim 1, wherein in the step 2), the protease is one or more of trypsin, pepsin, alkaline protease and papain.
5. The method for improving the emulsifying activity of perilla seed protein according to claim 1, wherein in the step 2), the pH value of the perilla seed meal mixed solution is adjusted to 2-11, and the temperature is adjusted to 35-65 ℃.
6. The method for improving the emulsifying activity of perilla seed protein according to claim 1, wherein in the step 3), the temperature of an air inlet of drying equipment for spray drying is set to be 150-220 ℃.
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| CN111406949A (en) * | 2019-12-04 | 2020-07-14 | 黑龙江万物生生物科技有限公司 | Whole perilla seed (meal) active peptide nutrient and preparation method thereof |
| CN111631297A (en) * | 2020-06-03 | 2020-09-08 | 陕西科技大学 | Stable microemulsion based on modified perilla seed protein and preparation method thereof |
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