CN107927319B - Method for preparing peanut polypeptide liquid by hydrolyzing peanut protein by using peanut inherent protease - Google Patents
Method for preparing peanut polypeptide liquid by hydrolyzing peanut protein by using peanut inherent protease Download PDFInfo
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/14—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from leguminous or other vegetable seeds; from press-cake or oil-bearing seeds
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- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
- A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
- A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
- A23J3/346—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of vegetable proteins
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/02—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation containing fruit or vegetable juices
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Abstract
A method for preparing peanut polypeptide liquid by hydrolyzing peanut protein by utilizing peanut inherent protease belongs to the technical field of processing of plant protein and grease. Peanut is soaked, red skin is removed, mildewed and deteriorated peanut kernels are removed, washing and grinding are carried out to prepare peanut pulp, a centrifuge is used for processing to obtain a peanut oil body enrichment substance and low-fat peanut pulp, peanut inherent protease in the low-fat peanut pulp is activated through acid processing, the activated peanut inherent protease hydrolyzes protein of the low-fat peanut pulp to generate low-fat peanut polypeptide liquid, and the peanut polypeptide liquid beverage can be obtained through fruit juice after blending and sterilization. The method does not use organic solvent and enzyme preparation, is an environment-friendly method with simple process and low processing cost, and can obtain peanut polypeptide liquid products with low fat, nutrition, health and good functionality; the peanut polypeptide liquid can be obtained, meanwhile, the byproduct peanut oil body enrichment can be obtained, and the peanut polypeptide liquid is a novel peanut oil product and can be used for soup bases and replacing thin butter and the like.
Description
Technical Field
The invention relates to a method for preparing peanut polypeptide liquid by hydrolyzing peanut protein by using peanut inherent protease, belonging to the technical field of processing of plant protein and grease.
Background
The production of polypeptide mainly includes two methods of synthesis and protein in vitro hydrolysis. The synthesis method comprises three methods of chemical synthesis, recombinant technology synthesis and enzymatic synthesis. Although the synthesis method can synthesize any active peptide at will, it has problems of high cost, many by-products, residues, and the like. In vitro methods of hydrolyzing proteins include acid hydrolysis, alkaline hydrolysis and enzymatic hydrolysis. The first two methods are simple and easy to operate, but may have harmful effects in nutrition and toxicology, so that the application thereof is limited to a certain extent. The enzymatic hydrolysis can carry out positioning hydrolysis under certain conditions to generate specific peptide, and the hydrolysis process is easy to control; generally, the loss of nutrition can not be caused, and the toxicological problem can not be caused. However, enzyme preparations are expensive, thus leading to a generally high price for current peptide products.
Peanuts are important oil crops and economic crops in China, the annual output is about 1500 ten thousand tons, and the peanuts contain about 25% of protein. At present, the main product form of the peanut protein is hot-pressed cake meal (or crushed cake meal) and cold-pressed cake meal, and the economic value is low. Peanut protein is a very characteristic plant protein, and is rich in arginine (about 13% of the total amino acids). Arginine has important effects on healthy reproduction, wound healing, ammonia discharge of human body, immunity improvement, blood pressure regulation and growth hormone generation promotion. Peanut protein is allergenic, especially for the European and American population. By the hydrolysis of the inherent protease of the peanut, not only can the anaphylaxis of the peanut protein be greatly reduced, but also the amino acid in the peanut protein can be more easily digested and absorbed; in addition, peanut intrinsic proteases hydrolyze peanut proteins to produce a variety of active polypeptide products. Therefore, the peanut inherent protease can be utilized to provide peanut polypeptide products with lower price and good functionality for the market.
Disclosure of Invention
The invention aims to provide a method for preparing peanut polypeptide liquid by hydrolyzing peanut protein by utilizing peanut inherent protease, provides a novel and high-added-value processing method for the peanut protein, and obtains a peanut polypeptide liquid product with good nutrition, health and functionality.
According to the technical scheme provided by the invention, the method for preparing the peanut polypeptide liquid by hydrolyzing peanut protein by utilizing the inherent protease of the peanut comprises the steps of soaking the peanut, removing red skin, removing mildew and deteriorated peanut kernels, leaching, grinding into slurry to prepare peanut slurry, then obtaining low-fat peanut slurry by a centrifugal machine, activating the inherent protease of the peanut by acid treatment, and hydrolyzing the low-fat peanut slurry protein by utilizing the activated inherent protease of the peanut to generate the peanut polypeptide liquid; blending and sterilizing to obtain peanut polypeptide liquid beverage; the method comprises the following specific steps:
(1) preparing peanut slurry: soaking unbaked peanut, removing red skin, manually selecting or color selecting to remove mildew and deteriorated peanut kernel, rinsing, grinding for 0.5-2min, and removing residue to obtain peanut pulp;
(2) repeating the extraction: adding water into the peanut residues obtained in the step (1), wherein the mass ratio of the peanut residues to the water is 1: 1-3, and grinding the mixture for 0.5-2min to obtain peanut pulp and peanut residues again; repeating the steps for 0-2 times, and combining the peanut pulp obtained in the step (1) and the peanut pulp obtained in the step (2);
(3) centrifugal degreasing: centrifuging the peanut pulp obtained in the step (2) at 3000-10000rpm for 30s-10min to obtain a layering liquid, wherein the heavy phase is low-fat peanut pulp; the light phase is the peanut oil body enrichment, namely the peanut oil part;
(4) peanut inherent protease activation: putting the low-fat peanut pulp obtained in the step (3) into an activation tank, adjusting the pH to 1-3 by using an acidic regulator, and then activating for 0.5-4h at the temperature of 20-60 ℃ to obtain the low-fat peanut pulp containing the activated peanut inherent protease;
(5) activating the inherent protease of the peanut to hydrolyze the peanut protein: and (4) putting the low-fat peanut pulp obtained in the step (4) into a hydrolysis tank, adjusting the pH to 2-5 by using an alkaline regulator, then reacting for 24-72h at 40-80 ℃, and obtaining peanut polypeptide liquid under the action of activated peanut endogenous protease.
The refining steps in the steps (1) and (2) are as follows: adding water into the artificially sorted or color-sorted peanuts, wherein the mass ratio of the peanuts to the water in the step (1) is 1: 1-9, wet-grinding for 0.5-2min, and filtering to obtain peanut pulp and peanut residues; and (3) continuously repeating the steps of adding water and grinding the peanut residues for 0-3 times, wherein the water adding amount is the mass ratio of the peanut residues to the water and is 1: 1-3, filtering, and combining the obtained pulp to obtain the peanut pulp.
And (4) the centrifugal machine used for centrifugal degreasing in the step (3) is a disk type centrifugal machine or a three-phase centrifugal machine.
The used peanut inherent protease is aspartic protease, and the activated aspartic protease has two polypeptide chains with the molecular weights of 32kDa and 9kDa respectively.
Activating aspartic protease by acid treatment; and (4) the acidity regulator is one of hydrochloric acid, citric acid, phosphoric acid or sulfuric acid.
Hydrolyzing the low-fat peanut pulp protein by using the activated peanut inherent protease to generate peanut polypeptide, wherein the alkaline regulator in the step (5) is one of soda, baking soda, sodium hydroxide or potassium hydroxide.
And (3) placing the obtained peanut polypeptide liquid into a blending tank, adding the ingredients of fructose, sucrose, maltose syrup and fruit juice, compounding, sterilizing and carrying out aseptic packaging to obtain the peanut polypeptide liquid beverage.
Sterilizing at 80-95 deg.C for 5-15 min.
The invention has the beneficial effects that: the method does not use organic solvent and industrial enzyme preparation, is an environment-friendly method with low processing cost, and can obtain the peanut polypeptide liquid product with good nutrition, health and functionality.
Drawings
FIG. 1 is an electrophoretogram of low-fat peanut slurry after 48h hydrolysis at pH2.5 after pH2 activation.
Detailed Description
In order to make the technical features, effects and advantages of the present invention more apparent, the present invention will be described in further detail with reference to the accompanying drawings.
Example 1
Soaking peanuts for 12h, removing red skins, removing mildewed and deteriorated peanut kernels, washing, adding water with the mass 6 times that of the peanut kernels, grinding the mixture into slurry for 2min, and filtering to obtain peanut slurry and peanut residues; centrifuging the peanut slurry at 9600rpm for 3min to obtain low-fat peanut slurry and peanut oil body concentrate; adjusting pH of the low-fat peanut pulp to 2 with hydrochloric acid, activating at 50 deg.C for 1h, adjusting pH to 2.5 with sodium bicarbonate, and keeping the temperature at 60 deg.C for 48h to obtain low-fat peanut polypeptide liquid; adding apple juice according to the ratio of apple juice/low-fat peanut polypeptide liquid =1/9, mixing, homogenizing, sterilizing (90 deg.C, 10 min), and packaging to obtain the beverage. The centrifuge is a three-phase centrifuge or a disk centrifuge.
Example 2
Soaking peanuts for 15h, removing red skins, removing mildewed and deteriorated peanut kernels, washing, adding water with the mass 4 times that of the peanut kernels, grinding the peanut kernels for 2min, and filtering to obtain peanut pulp I and peanut residues I; adding water into peanut residue I at a mass ratio of 1: 3, and grinding for 1min to obtain peanut pulp II and peanut residue II; mixing the peanut pulp I and the peanut pulp II to obtain peanut pulp III; centrifuging the peanut slurry III at 7000rpm for 3min to obtain low-fat peanut slurry and peanut oil body concentrate; adjusting the pH value of the low-fat peanut pulp to 1.8 by using phosphoric acid, and activating for 2 hours at the temperature of 40 ℃; adjusting pH to 2.3 with sodium carbonate, and keeping the temperature at 55 deg.C for 55 hr to obtain low-fat peanut polypeptide liquid; adding peach juice according to the proportion of peach juice/low-fat peanut polypeptide liquid =1/9, mixing, homogenizing, sterilizing (95 deg.C, 5 min), and packaging to obtain the beverage. The centrifuge is a three-phase centrifuge or a disk centrifuge.
Example 3
Soaking peanuts for 15h, removing red skins, removing mildewed and deteriorated peanut kernels, washing, adding water with the mass 6 times that of the peanut kernels, grinding the mixture into slurry for 2min, and filtering to obtain peanut slurry I and peanut residues I; adding water into peanut residue I at a mass ratio of 1: 3, and grinding for 1.5min to obtain peanut pulp II and peanut residue II; mixing the peanut pulp I and the peanut pulp II to obtain peanut pulp III; centrifuging the peanut slurry III at 5000rpm for 5min to obtain low-fat peanut slurry and peanut oil body concentrate; adjusting the pH value of the low-fat peanut pulp to 3 by using hydrochloric acid, and activating for 1.5h at the temperature of 60 ℃; keeping the temperature at 55 ℃ for 60h without adjusting the pH value to obtain low-fat peanut polypeptide liquid; adding guava juice according to the ratio of guava juice/low-fat peanut polypeptide liquid =1/9, mixing, homogenizing, sterilizing (80 deg.C, 15 min), and packaging to obtain guava juice peanut polypeptide liquid beverage. The centrifuge is a three-phase centrifuge or a disk centrifuge.
Example 4
Soaking peanuts for 9h, removing red skins, removing mildewed and deteriorated peanut kernels, cleaning, adding water with the mass 5 times that of the peanut kernels, grinding the mixture into slurry for 2min, and filtering to obtain peanut slurry I and peanut residues I; adding water into peanut residues I, wherein the mass ratio of the peanut residues I to the water is 1: 2, and grinding for 30s to obtain peanut pulp II and peanut residues II; adding water into peanut residues II at a mass ratio of 1: 2, and grinding for 30s to obtain peanut pulp III and peanut residues III; mixing the peanut slurry I, II and the peanut slurry III to obtain a peanut slurry IV; centrifuging the peanut pulp IV for 1min at 9600rpm to obtain low-fat peanut pulp and a peanut oil body enrichment; adjusting pH of the low-fat peanut pulp to 2 with phosphoric acid, and activating at 60 deg.C for 0.5 h; keeping the temperature at 60 ℃ for 36h without adjusting the pH value to obtain low-fat peanut polypeptide liquid; adding litchi juice according to the ratio of litchi juice/low-fat peanut polypeptide liquid =1/9, mixing, homogenizing, sterilizing (85 ℃ for 10 min), and packaging to obtain the litchi juice peanut polypeptide liquid beverage. The centrifuge is a three-phase centrifuge or a disk centrifuge.
Application example 1 evaluation of Low fat peanut polypeptide liquid
The analysis method of the main evaluation index is as follows:
tris (hydroxymethyl) methylglycine-sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The concentrations of the prepared concentrated gel and the separation gel were 4% and 16%, the protein concentration of the sample was diluted to 2mg/mL, and 1/1 was added to a sample dissolving solution (4% sodium dodecyl sulfate, 20% glycerol, 0.125M tris (hydroxymethyl) aminomethane-hydrochloric acid buffer (pH 6.8), 0.01% bromophenol blue) to make the protein concentration to 1 mg/mL. Adding 2% (v/v) mercaptoethanol, heating in boiling water bath for 3min, and centrifuging at 10000rpm for 5 min. The sample adding amount is 10 mu L, the voltage is set to be 30mV at the beginning, the voltage is adjusted to be 100mV after all the samples enter the concentrated gel, the constant voltage is kept until the samples reach the bottom of the gel, and the electrophoresis is finished. After completion, the gel was removed and stained with Coomassie blue G-250.
Acid hydrolysis amino acid analysis. Adding the low-fat peanut polypeptide liquid into a hydrolysis tube, adding concentrated hydrochloric acid to make the concentration of hydrochloric acid be 6M, filling nitrogen, and sealing the tube. After hydrolysis at 120 ℃ for 22h, neutralization was carried out with sodium hydroxide, double-layer filter paper filtration, centrifugation (15000 rpm, 30 min) and filling into sample bottles. The measuring equipment is an Agilent 1100 and an Agilent 1260 amino acid special high performance liquid chromatograph.
After 48h of hydrolysis (FIG. 1) under the preferred conditions (pH 2 activation, pH2.5 hydrolysis), 66kDa peanut protein (allergenic protein), 45kDa peanut protein and 35kDa peanut protein are substantially hydrolyzed by the activated intrinsic protease, with a partial residue of 22kDa peanut protein; the <22kDa polypeptide component accounts for more than half of the total protein.
Amino acid composition of low fat peanut polypeptide liquid. After hydrolysis under the preferred conditions (pH 2 activation, pH2.5 hydrolysis) for 48h, the amino acid composition of the reduced fat peanut polypeptide liquid was analyzed (Table 1). The 100g low-fat peanut slurry polypeptide liquid contains 1.9563g amino acids, wherein the arginine accounts for 0.2491g and accounts for 13 percent of the total acid hydrolysis amino acids. It can be seen that arginine is a major amino acid in low fat peanut polypeptide fluids.
TABLE 1 amino acid composition of low fat peanut pulp (g/100 g of low fat peanut polypeptide liquid) after 48h hydrolysis at pH2.5 after pH2 activation.
TABLE 1 amino acid composition (acid hydrolysis) of low fat peanut polypeptide liquid
Use example 2 flavoring of Low fat peanut polypeptide liquid
The low-fat peanut polypeptide liquid is sour in taste, has light peanut flavor and needs seasoning treatment. In order to improve the taste, the invention carries out seasoning treatment by using fruit juice, and separately considers guava juice, apple juice, pear juice, peach juice, lychee juice, orange juice, kiwi fruit juice, grapefruit juice, mango juice and strawberry juice. Seasoning according to the low fat peanut polypeptide liquid/juice =9/1 volume ratio, the results show (table 2): the guava juice, the apple juice, the peach juice, the pear juice and the litchi juice can weaken the sour taste of the polypeptide liquid and endow the final beverage with the flavor of the juice; three juices, namely grapefruit juice, orange juice and kiwi fruit juice, are sour and bitter in taste, are simply used for seasoning, are poor in taste, and can be accepted after cane sugar with the mass percentage of 3% is added; the taste of the mango juice is strange after the mango juice is added, and simultaneously, the mango juice generates astringent taste and is unacceptable; the strawberry juice has a normal taste after being added. It should be noted that, the higher the ratio of the guava juice, the apple juice, the peach juice, the pear juice and the litchi juice, the better the taste of the final beverage.
TABLE 2 flavoring effect of different juices on low fat peanut polypeptide liquids
Note: fruit juice/low fat peanut polypeptide liquid = 1/9.
Finally, it should be noted that: although the present invention has been described in detail with reference to the above embodiments, it should be understood by those skilled in the art that: modifications and equivalents may be made thereto without departing from the spirit and scope of the invention and it is intended to cover in the claims the invention as defined in the appended claims.
Claims (8)
1. A method for preparing peanut polypeptide liquid by hydrolyzing peanut protein by using peanut inherent protease is characterized by comprising the following steps: soaking peanuts, removing red skins, removing mildewed and deteriorated peanut kernels, leaching, grinding into slurry to prepare peanut slurry, then obtaining low-fat peanut slurry through a centrifuge, activating inherent protease of the peanuts through acid treatment, and hydrolyzing the protein of the low-fat peanut slurry by using the activated inherent protease of the peanuts to generate peanut polypeptide liquid; the method comprises the following specific steps:
(1) preparing peanut slurry: soaking unbaked peanut, removing red skin, manually selecting or color selecting to remove mildew and deteriorated peanut kernel, rinsing, grinding for 0.5-2min, and removing residue to obtain peanut pulp;
(2) repeating the extraction: adding water into the peanut residues obtained in the step (1), wherein the mass ratio of the peanut residues to the water is 1: 1-3, and grinding the mixture for 0.5-2min to obtain peanut pulp and peanut residues again; repeating the steps for 0-2 times, and combining the peanut pulp obtained in the step (1) and the peanut pulp obtained in the step (2);
(3) centrifugal degreasing: centrifuging the peanut pulp obtained in the step (2) at 3000-10000rpm for 30s-10min to obtain a layering liquid, wherein the heavy phase is low-fat peanut pulp; the light phase is the peanut oil body enrichment, namely the peanut oil part;
(4) peanut inherent protease activation: putting the low-fat peanut pulp obtained in the step (3) into an activation tank, adjusting the pH to 1-3 by using an acidic regulator, and then activating for 0.5-4h at the temperature of 20-60 ℃ to obtain the low-fat peanut pulp containing the activated peanut inherent protease;
(5) activating the inherent protease of the peanut to hydrolyze the peanut protein: and (4) putting the low-fat peanut pulp obtained in the step (4) into a hydrolysis tank, adjusting the pH to 2-5 by using an alkaline regulator, then reacting for 24-72h at 40-80 ℃, and obtaining peanut polypeptide liquid under the action of activated peanut endogenous protease.
2. The method for preparing peanut polypeptide liquid by hydrolyzing peanut protein with peanut inherent protease as claimed in claim 1, wherein the refining steps in steps (1) and (2) are as follows: adding water into the artificially sorted or color-sorted peanuts, wherein the mass ratio of the peanuts to the water in the step (1) is 1: 1-9, wet-grinding for 0.5-2min, and filtering to obtain peanut pulp and peanut residues; and (3) continuously repeating the steps of adding water and grinding the peanut residues for 0-3 times, wherein the water adding amount is the mass ratio of the peanut residues to the water and is 1: 1-3, filtering, and combining the obtained pulp to obtain the peanut pulp.
3. The method of claim 1, wherein the peanut polypeptide solution is prepared by hydrolyzing peanut protein with peanut inherent protease, wherein the method comprises the following steps: and (4) the centrifugal machine used for centrifugal degreasing in the step (3) is a disk type centrifugal machine or a three-phase centrifugal machine.
4. The method of claim 1, wherein the peanut polypeptide solution is prepared by hydrolyzing peanut protein with peanut inherent protease, wherein the method comprises the following steps: the used peanut inherent protease is aspartic protease, and the activated aspartic protease has two polypeptide chains with the molecular weights of 32kDa and 9kDa respectively.
5. The method of claim 4, wherein the peanut polypeptide solution is prepared by hydrolyzing peanut protein with peanut inherent protease, wherein the method comprises the following steps: activating aspartic protease by acid treatment; and (4) the acidity regulator is one of hydrochloric acid, citric acid, phosphoric acid or sulfuric acid.
6. The method of claim 1, wherein the peanut polypeptide solution is prepared by hydrolyzing peanut protein with peanut inherent protease, wherein the method comprises the following steps: hydrolyzing the low-fat peanut pulp protein by using the activated peanut inherent protease to generate peanut polypeptide, wherein the alkaline regulator in the step (5) is one of soda, baking soda, sodium hydroxide or potassium hydroxide.
7. The method of claim 1, wherein the peanut polypeptide solution is prepared by hydrolyzing peanut protein with peanut inherent protease, wherein the method comprises the following steps: and (3) placing the obtained peanut polypeptide liquid into a blending tank, adding the ingredients of fructose, sucrose, maltose syrup and fruit juice, compounding, sterilizing and carrying out aseptic packaging to obtain the peanut polypeptide liquid beverage.
8. The method of claim 7, wherein the peanut polypeptide solution is prepared by hydrolyzing peanut protein with peanut inherent protease, wherein the method comprises: sterilizing at 80-95 deg.C for 5-15 min.
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| CN1635136A (en) * | 2003-12-30 | 2005-07-06 | 北京市食品研究所 | Method for preparing plant separated protein |
| CN106929141A (en) * | 2017-03-28 | 2017-07-07 | 江南大学 | A kind of method that peanut oil is prepared based on peanut endogenous protease water law demulsification peanut oil body |
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| WO2013010037A1 (en) * | 2011-07-12 | 2013-01-17 | Lyrical Foods, Inc. | Methods and compositions for consumables |
| WO2016005587A1 (en) * | 2014-07-11 | 2016-01-14 | Chr. Hansen A/S | Milk clotting aspartic protease enzyme composition |
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| CN1635136A (en) * | 2003-12-30 | 2005-07-06 | 北京市食品研究所 | Method for preparing plant separated protein |
| CN106929141A (en) * | 2017-03-28 | 2017-07-07 | 江南大学 | A kind of method that peanut oil is prepared based on peanut endogenous protease water law demulsification peanut oil body |
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