CN113208118A - Preparation method of low-bitter-taste antioxidant peptide beverage - Google Patents
Preparation method of low-bitter-taste antioxidant peptide beverage Download PDFInfo
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- CN113208118A CN113208118A CN202110665358.6A CN202110665358A CN113208118A CN 113208118 A CN113208118 A CN 113208118A CN 202110665358 A CN202110665358 A CN 202110665358A CN 113208118 A CN113208118 A CN 113208118A
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Abstract
A method for preparing a low-bitter antioxidant peptide beverage comprises: (1) the method comprises the following steps of (1) carrying out first-step enzymolysis on soybean protein isolate, (2) carrying out second-step enzymolysis on enzymolysis liquid, (3) processing and preparing antioxidant peptides, and (4) preparing a low-bitter antioxidant peptide beverage. The invention exposes hydrophobic groups in the isolated soy protein molecules through first-step enzymolysis, thereby generating stronger bitter taste, then carries out second-step enzymolysis, obviously reduces the threshold range of the bitter taste after the second-step enzymolysis, which is consistent with the expected goal of reaching low bitter taste, and then can reach novel products with high nutrition and low bitter taste which are easy to accept through the modulation of a formula.
Description
Technical Field
The invention belongs to the technical field of functional food processing, and relates to a preparation method of a low-bitter antioxidant peptide beverage.
Background
Because of its unique chemical composition, soybean is one of the most economically valuable and useful agricultural products in the world. In cereals and other legumes, the protein content in soybeans is highest, about 40%. The protein obtained by using low-temperature defatted soybean meal as a raw material and adopting a centrifugal separation technology is called soybean protein isolate. The soybean protein isolate is a high-quality plant protein, has high protein concentration and balanced nutrient components, is rich in amino acid necessary for a human body, and has high digestion and utilization rate in the human body. The proper addition of soy protein isolate in the food product can improve the protein efficiency ratio. Soy protein isolates are classified into 2S, 7S, 11S and 15S proteins according to the rate of protein sedimentation, with the two components with the highest content being the 7S and 11S proteins.
The soybean peptide is a mixture obtained by treating soybean protein after protease action, and comprises peptides with different molecular chains as main components, free amino acids and a small amount of saccharides. The soybean peptide has a molecular weight distribution of less than 1000, is balanced in essential amino acids and rich in content, not only has processing characteristics richer than soybean protein, such as no beany flavor, is not easy to denature and precipitate under the conditions of heat, acid and alkali, has good foaming characteristics and hygroscopicity, has a gel softening function, has the functions of promoting the proliferation of microorganisms and promoting the generation of beneficial metabolites, and also has a plurality of important treatment characteristics, such as the characteristic of easy digestion and absorption of the soybean peptide, the capability of promoting fat metabolism, enhancing muscle strength, helping to recover fatigue and enhance physical strength in time, and simultaneously has low antigenicity, cholesterol reduction, blood pressure increase inhibition and the like, and the soybean peptide is more and more valued as a very useful functional food raw material. By utilizing these characteristics of soybean peptide, people can develop it into functional foods, athlete foods, foods for special patients, or apply it as a food material having good processing characteristics to foods and related fields.
Improving the functional properties of soy protein and increasing the bioavailability of the nutritional components of soy products is a problem to be solved in the food industry. Although the basic properties of the soybean protein can be changed by common methods such as physical modification and chemical modification, a series of problems are brought, the investment is large, the yield is small, the safety and the like are not negligible, and after the soybean protein is subjected to enzymolysis to form the soybean peptide by using incomplete hydrolysis of enzyme, a plurality of original functional properties of the soybean protein, such as an emulsifying function, a foaming function, a water absorption function, a thickening function, an adhesion function, a nutritional value, a physiological function and the like, are improved to different degrees, so that the requirements of the food industry and related industries can be better met.
The soybean protein hydrolysate has small molecular weight, high water solubility, no protein denaturation, and easy digestion and absorption in human body, and is especially suitable for production of functional, health-care and curative food, clinical nutritive product, instant beverage and instant food. However, the hydrolysate often contains a small amount of free amino acids such as leucine and methionine, derivatives thereof and small molecular hydrophobic peptide substances which can generate bitter taste, so that the product has different bitter taste, the flavor of food is directly influenced, some troubles are brought to people to eat, and the popularization and application of the product in the food industry are limited to a certain extent. We further processed the bitter tasting substance by a fractional enzymatic hydrolysis process to produce a low bitter soy polypeptide.
Disclosure of Invention
The invention aims to provide a preparation method of a low-bitter antioxidant peptide beverage.
In order to achieve the above objects and other related objects, the present invention provides the following technical solutions: a method for preparing a low-bitter antioxidant peptide beverage comprises the following steps:
step 1: dispersing the soybean protein isolate in distilled water to obtain a mixed solution with the mass concentration of 3-7%, heating in a water bath at the temperature of 80-90 ℃ for 25-35min to modify the soybean protein isolate, then cooling to room temperature, adding alkaline protease, carrying out enzymolysis for 3-5 h at the temperature of 40-70 ℃, and obtaining a first-step enzymolysis liquid after the enzymolysis is finished;
step 2: carrying out boiling water bath enzyme deactivation on the first-step enzymolysis liquid, adding pepsin for continuous enzymolysis, carrying out enzymolysis for 3-5 hours at the temperature of 40-60 ℃, and obtaining second-step enzymolysis liquid after the enzymolysis is finished;
and step 3: carrying out enzyme deactivation on the enzymolysis liquid in the second step, filtering, taking filtrate, carrying out rotary evaporation to obtain a crude product, freeze-drying the crude product to obtain polypeptide powder, and grinding and sieving by a 60-mesh sieve to obtain the low-bitter-taste antioxidant peptide;
and 4, step 4: the formula of the low-bitter antioxidant peptide beverage comprises the following raw materials in percentage by mass: 1.5% of low-bitter-taste antioxidant peptide, 4.5-5.5% of white granulated sugar, 0.1-0.15% of citric acid, 0.055-0.75% of sodium citrate, 0.055-0.75% of malic acid, 0.04-0.06% of lactic acid, 0-0.05% of essence and the balance of purified water;
mixing the materials at 65-85 deg.C, stirring, and homogenizing under 30-40 MPa; and sterilizing the material after high-pressure homogenization, and aseptically filling to obtain the low-bitter antioxidant peptide beverage product.
The preferable technical scheme is as follows: in the step 1, the substrate ratio is 500-1000U/g during enzymolysis.
The preferable technical scheme is as follows: in the step 2, during enzymolysis, the substrate ratio is 500-1000U/g.
Due to the application of the technical scheme, compared with the prior art, the invention has the advantages that:
1. the invention can achieve the aim of modifying the isolated soy protein by utilizing a stepwise enzymolysis method, is more suitable compared with other modification methods, and can improve the application of the soy protein. The development of antioxidant soybean peptide can greatly improve the added value of soybean products and broaden the application of soybeans. Therefore, the research and development of the soybean antioxidant peptide have wide prospect and certain social and economic benefits.
2. The invention researches and develops the obtained antioxidant peptide and successfully develops a low-bitter beverage rich in soybean protein peptide, achieves the aim from research to practical application, fully proves the feasibility of the beverage, and provides a new idea for comprehensively utilizing the soybean protein isolate to a greater extent.
3. The invention exposes hydrophobic groups in the isolated soy protein molecules through first-step enzymolysis, thereby generating stronger bitter taste, then carries out second-step enzymolysis, obviously reduces the threshold range of the bitter taste after the second-step enzymolysis, which is consistent with the expected goal of reaching low bitter taste, and then can reach novel products with high nutrition and low bitter taste which are easy to accept through the modulation of a formula.
Drawings
FIG. 1 is a graph showing the effect of soy protein isolate concentration on degree of hydrolysis
FIG. 2 shows the effect of substrate specific concentration on the degree of hydrolysis.
FIG. 3 is a graph showing the effect of the temperature of the enzymatic hydrolysis on the degree of hydrolysis.
FIG. 4 is a graph showing the effect of the second step of enzymatic digestion on bitterness score.
FIG. 5 is a graph of the antioxidant activity of polypeptides.
Detailed Description
The following description of the embodiments of the present invention is provided for illustrative purposes, and other advantages and effects of the present invention will become apparent to those skilled in the art from the present disclosure.
Please refer to fig. 1 to 5. It should be understood that in the description of the present invention, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc. indicate the orientation or positional relationship based on the orientation or positional relationship shown in the drawings, or the orientation or positional relationship which the product of the present invention is usually placed in when used, which is only for the convenience of describing the present invention and simplifying the description, but does not indicate or imply that the device or element referred to must have a specific orientation, be constructed in a specific orientation, and be operated, and therefore, should not be construed as limiting the present invention. Furthermore, the terms "first," "second," "third," and the like are used solely to distinguish one from another and are not to be construed as indicating or implying relative importance. The terms "horizontal", "vertical", "overhang" and the like do not imply that the components are required to be absolutely horizontal or overhang, but may be slightly inclined. For example, "horizontal" merely means that the direction is more horizontal than "vertical" and does not mean that the structure must be perfectly horizontal, but may be slightly inclined.
Example 1: preparation method of low-bitter-taste antioxidant peptide beverage
A preparation method of a low-bitter antioxidant peptide beverage is characterized by comprising the following technical steps.
(1) First step enzymolysis of soy protein isolate
Weighing a proper amount of soybean protein isolate, dissolving the soybean protein isolate in distilled water, controlling the concentration of the soybean protein isolate to be 3-7%, heating the soybean protein isolate in water bath at 85 ℃ for 30min after the soybean protein isolate is fully dissolved, modifying the soybean protein isolate, cooling the soybean protein isolate to room temperature, adding a proper amount of alkaline protease (Alcalase) for enzymolysis, controlling the substrate ratio to be 500-1000U/g, controlling the temperature to be 40-70 ℃ for enzymolysis for 3-5 h, and obtaining a first-step enzymolysis liquid after the enzymolysis is finished.
(2) Second step enzymolysis of the enzymolysis liquid
And (3) after carrying out boiling water bath 5min enzyme deactivation on the first-step enzymolysis liquid, adding aeolian pepsin for continuous enzymolysis, controlling the substrate ratio at 500-1000U/g, the temperature at 40-60 ℃, controlling the enzymolysis time at 3-5 h, and collecting the second-step enzymolysis liquid after completing the enzymolysis.
(3) Processing preparation of antioxidant peptide
And (2) carrying out enzyme deactivation operation on the collected enzymatic hydrolysate, then carrying out suction filtration (the size of the filter paper is 11 x 11cm, the aperture of the filter paper is 0.02 mm), taking supernatant fluid to carry out rotary evaporation at 60 ℃ under 0.095MPa to obtain thick liquid, freeze-drying (20 MPa and 40 ℃ below zero for 24 hours) the crude product, repeatedly freeze-drying until the product is completely dried to obtain final polypeptide powder, namely the soybean protein polypeptide, grinding, and sieving with a 60-mesh sieve to obtain the required antioxidant peptide.
(4) Method for preparing low-bitter antioxidant peptide beverage
The beverage formula is as follows: 1.5 percent (3) of low-bitterness antioxidant peptide, 5.0 percent of white granulated sugar, 0.125 percent of citric acid, 0.065 percent of sodium citrate, 0.065 percent of malic acid, 0.05 percent of lactic acid, 0-0.05 percent of essence and the balance of food-grade purified water.
Mixing the materials at 65-85 deg.C, stirring, homogenizing under 30-40MPa, and processing for 2-4 times.
UHT sterilization is adopted for the material after high-pressure homogenization, the sterilization temperature is 130 ℃ and 140 ℃, and the sterilization time is 10-15 s; then cooling to a temperature lower than 40 ℃ within 30-45min, and aseptically filling to obtain the low-bitter-taste antioxidant peptide beverage product.
The pH-star method is simple to operate and can be used for continuous determination, so that the method is adopted for determining the hydrolysis degree. Namely, firstly, recording the pH value under the initial stable condition, then hydrolyzing the isolated soy protein under the alkaline condition, reducing the pH value by the released amino acid, titrating the isolated soy protein back to the initial pH value by using dilute alkali liquor, knowing the breaking condition of protein peptide bonds, and calculating according to a formula.
DH=B×Nb× α-1×MP-1× hhot-1×100%；
In the formula: DH is degree of hydrolysis,%; b is the volume mL of alkali liquor; n is a radical ofbIs the concentration of alkali liquor, mol/L; α is the average degree of amino dissociation; mP Is the protein content in the substrate, g; h ishotIs the molarity of peptide bonds in proteins, mmol/g.
The higher the degree of hydrolysis, i.e.the more thorough the hydrolysis of the amino acid chain, the more complete the hydrolysis, the higher the polypeptide content. With the change of the concentration of the soy protein isolate, the optimal conditions for preparing the polypeptide can be selected under the optimal concentration. The results are shown in FIG. 1.
The degree of hydrolysis was measured by the pH-star method. Namely, firstly, recording the pH value under the initial stable condition, then hydrolyzing the isolated soy protein under the alkaline condition, reducing the pH value by the released amino acid, titrating the isolated soy protein back to the initial pH value by using dilute alkali liquor, knowing the breaking condition of protein peptide bonds, and calculating according to a formula.
DH=B×Nb× α-1×MP-1× hhot-1×100%；
In the formula: DH is degree of hydrolysis,%; b is the volume mL of alkali liquor; n is a radical ofbIs the concentration of alkali liquor, mol/L; α is the average degree of amino dissociation; mPIs the protein content in the substrate, g; h ishotIs the molarity of peptide bonds in proteins, mmol/g.
By exploring the substrate specific concentration and taking the result of the enzymolysis time as an index, the optimal substrate specific concentration can be optimized, and the optimal hydrolysis effect can be achieved under the condition. In order to prepare the desired polypeptide. The results are shown in FIG. 2.
The degree of hydrolysis was measured by the pH-star method. Namely, firstly, recording the pH value under the initial stable condition, then hydrolyzing the isolated soy protein under the alkaline condition, reducing the pH value by the released amino acid, titrating the isolated soy protein back to the initial pH value by using dilute alkali liquor, knowing the breaking condition of protein peptide bonds, and calculating according to a formula.
DH=B×Nb× α-1×MP-1× hhot -1×100%；
In the formula: DH is degree of hydrolysis,%; b is the volume mL of alkali liquor; n is a radical ofbIs the concentration of alkali liquor, mol/L; α is the average degree of amino dissociation; mPIs the protein content in the substrate, g; h ishotIs the molarity of peptide bonds in proteins, mmol/g.
By exploring the enzymolysis temperature and taking the result of the enzymolysis time as an index, the optimal substrate specific concentration can be optimized, and the optimal hydrolysis effect can be achieved under the condition. In order to prepare the desired polypeptide. The results are shown in FIG. 3.
0.19 of quinoline was weighed and dissolved in 50mL of 5% ethanol solution, and 5mL of the solution was diluted to 100 mL with distilled water to obtain 1.0X 10-4g/mL of the quinine solution, and further diluting to 5X 10-5、2.5×10-5、1×10-5、5×10-6g/mL of equiconcentrated quinline solution with bitterness values of 10, 5, 2.5, 1 and 0.5 respectively, and comparing the prepared soybean polypeptide with different concentrations of quinline solution to determine the bitterness of the soybean polypeptideAnd (4) taste value. After the second step of enzymatic hydrolysis, as shown in FIG. 4, it is clearly seen that the bitterness value of the polypeptide decreases with increasing time of enzymatic hydrolysis, which is beneficial for the application of the polypeptide in the food industry.
DPPH, 1-diphenyl-2-trinitrophenylhydrazine, was used for in vitro antioxidant evaluation of antioxidant ingredients. The detection method comprises the following steps: 2mL of each sample was prepared at different concentrations of 1, 3, 5, 7, 9 and 10 mg/mL. To each concentration was added 2.5mL of a 100. mu.L/LDPPH-methanol solution, and allowed to react at room temperature for 30 minutes, followed by absorbance measurement at 517nm (A)1). The samples were then replaced by deionized water and DPPH, and the absorbances under the same conditions were respectively expressed as A0And A2。
DPPH radical clearance: i% = (a)0-A1 + A2）/ A0 × 100％。
As shown in FIG. 5, the result of the DPPH free radical scavenging assay is analyzed, and the antioxidant activity of the polypeptide is obviously improved along with the increase of the concentration of the polypeptide, so that the polypeptide has excellent antioxidant activity.
Example 2: preparation method of low-bitter-taste antioxidant peptide beverage
A method for preparing a low-bitter antioxidant peptide beverage comprises the following steps:
step 1: dispersing the isolated soy protein in distilled water to obtain a mixed solution with the mass concentration of 5%, heating in a water bath at 85 ℃ for 30min to modify the isolated soy protein, then cooling to room temperature, adding alkaline protease, performing enzymolysis at 55 ℃ for 4h, and obtaining a first-step enzymolysis solution after the enzymolysis is finished;
step 2: carrying out boiling water bath enzyme deactivation on the first-step enzymolysis liquid, adding pepsin for continuous enzymolysis, carrying out enzymolysis for 4 hours at the temperature of 50 ℃, and obtaining second-step enzymolysis liquid after the enzymolysis is finished;
and step 3: carrying out enzyme deactivation on the enzymolysis liquid in the second step, filtering, taking filtrate, carrying out rotary evaporation to obtain a crude product, freeze-drying the crude product to obtain polypeptide powder, and grinding and sieving by a 60-mesh sieve to obtain the low-bitter-taste antioxidant peptide;
and 4, step 4: the formula of the low-bitter antioxidant peptide beverage comprises the following raw materials in percentage by mass: 1.5% of low-bitter-taste antioxidant peptide, 5% of white granulated sugar, 0.125% of citric acid, 0.065% of sodium citrate, 0.065% of malic acid, 0.05% of lactic acid and the balance of purified water;
mixing various materials according to a formula at 75 ℃, uniformly stirring, and homogenizing the uniformly mixed materials under high pressure of 35 MPa; and sterilizing the material after high-pressure homogenization, and aseptically filling to obtain the low-bitter antioxidant peptide beverage product.
The preferred embodiment is: in the step 1, the substrate ratio is 800U/g during enzymolysis.
The preferred embodiment is: in the step 2, the substrate ratio is 800U/g during enzymolysis.
Example 3: preparation method of low-bitter-taste antioxidant peptide beverage
A method for preparing a low-bitter antioxidant peptide beverage comprises the following steps:
step 1: dispersing the soybean protein isolate in distilled water to obtain a mixed solution with the mass concentration of 3%, heating in a water bath at 80 ℃ for 25min to modify the soybean protein isolate, then cooling to room temperature, adding alkaline protease, performing enzymolysis at 40 ℃ for 3-5 h to obtain a first-step enzymolysis solution;
step 2: carrying out boiling water bath enzyme deactivation on the first-step enzymolysis liquid, adding pepsin for continuous enzymolysis, carrying out enzymolysis for 3 hours at the temperature of 40 ℃, and obtaining second-step enzymolysis liquid after the enzymolysis is finished;
and step 3: carrying out enzyme deactivation on the enzymolysis liquid in the second step, filtering, taking filtrate, carrying out rotary evaporation to obtain a crude product, freeze-drying the crude product to obtain polypeptide powder, and grinding and sieving by a 60-mesh sieve to obtain the low-bitter-taste antioxidant peptide;
and 4, step 4: the formula of the low-bitter antioxidant peptide beverage comprises the following raw materials in percentage by mass: 1.5% of low-bitter-taste antioxidant peptide, 4.5% of white granulated sugar, 0.1% of citric acid, 0.055% of sodium citrate, 0.055% of malic acid, 0.04% of lactic acid, 0.05% of essence and the balance of purified water;
mixing various materials according to a formula at 65 ℃, uniformly stirring, and homogenizing the uniformly mixed materials under a high pressure of 300 MPa; and sterilizing the material after high-pressure homogenization, and aseptically filling to obtain the low-bitter antioxidant peptide beverage product.
The preferred embodiment is: in the step 1, the substrate ratio is 500U/gU/g during enzymolysis.
The preferred embodiment is: in the step 2, the substrate ratio is 500U/gU/g during enzymolysis.
The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit 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 (3)
1. A preparation method of a low-bitter antioxidant peptide beverage is characterized by comprising the following steps: comprises the following steps:
step 1: dispersing the soybean protein isolate in distilled water to obtain a mixed solution with the mass concentration of 3-7%, heating in a water bath at the temperature of 80-90 ℃ for 25-35min to modify the soybean protein isolate, then cooling to room temperature, adding alkaline protease, carrying out enzymolysis for 3-5 h at the temperature of 40-70 ℃, and obtaining a first-step enzymolysis liquid after the enzymolysis is finished;
step 2: carrying out boiling water bath enzyme deactivation on the first-step enzymolysis liquid, adding pepsin for continuous enzymolysis, carrying out enzymolysis for 3-5 hours at the temperature of 40-60 ℃, and obtaining second-step enzymolysis liquid after the enzymolysis is finished;
and step 3: carrying out enzyme deactivation on the enzymolysis liquid in the second step, filtering, taking filtrate, carrying out rotary evaporation to obtain a crude product, freeze-drying the crude product to obtain polypeptide powder, and grinding and sieving by a 60-mesh sieve to obtain the low-bitter-taste antioxidant peptide;
and 4, step 4: the formula of the low-bitter antioxidant peptide beverage comprises the following raw materials in percentage by mass: 1.5% of low-bitter-taste antioxidant peptide, 4.5-5.5% of white granulated sugar, 0.1-0.15% of citric acid, 0.055-0.75% of sodium citrate, 0.055-0.75% of malic acid, 0.04-0.06% of lactic acid, 0-0.05% of essence and the balance of purified water;
mixing the materials at 65-85 deg.C, stirring, and homogenizing under 30-40 MPa; and sterilizing the material after high-pressure homogenization, and aseptically filling to obtain the low-bitter antioxidant peptide beverage product.
2. The method of making a low bitter antioxidant peptide beverage of claim 1, wherein: in the step 1, the substrate ratio is 500-1000U/g during enzymolysis.
3. The method of making a low bitter antioxidant peptide beverage of claim 1, wherein: in the step 2, during enzymolysis, the substrate ratio is 500-1000U/g.
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