CN112690366A - Preparation method and application of rice protein peptide - Google Patents
Preparation method and application of rice protein peptide Download PDFInfo
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- CN112690366A CN112690366A CN202011573464.3A CN202011573464A CN112690366A CN 112690366 A CN112690366 A CN 112690366A CN 202011573464 A CN202011573464 A CN 202011573464A CN 112690366 A CN112690366 A CN 112690366A
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- 235000009566 rice Nutrition 0.000 title claims abstract description 178
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- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 91
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- 206010020751 Hypersensitivity Diseases 0.000 description 1
- 206010022998 Irritability Diseases 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
Abstract
The invention belongs to the technical field of rice protein peptide preparation, and particularly relates to a preparation method and application of rice protein peptide. The preparation method of the rice protein peptide provided by the invention is mainly characterized in that rice is dried, then a screw extruder is adopted to extrude the rice, and then the rice is homogenized under high pressure and subjected to enzymolysis reaction, thus obtaining the rice protein peptide. The preparation method of the rice protein peptide provided by the invention effectively solves the problem that strong alkali damages proteins during alkaline extraction, avoids the generation of toxic substances and bitter and astringent peculiar smell caused by condensation between amino acids, has high purity and solubility, is beneficial to the absorption of animals on the proteins, has the extraction rate as high as 85%, and can effectively save the cost.
Description
Technical Field
The invention belongs to the technical field of rice protein peptide preparation, and particularly relates to a preparation method and application of rice protein peptide.
Background
The rice is one of the most abundant grain crops in the global yield, and the rice protein is used as a product processed by the rice, has abundant resources and balanced nutrition and is not easy to generate allergy. Rice protein refers to the protein fraction of rice grains. Generally, rice contains about 8% of protein. The rice protein can be divided into four major categories according to the solubility difference of each component in the rice protein, namely, albumin which can dissolve in water and accounts for 2 to 5 percent of the total amount; secondly, the globulin is dissolved in 0.5mol/L NaCl solution, accounting for 2 to 10 percent of the total amount; ③ gluten, protein dissolved in dilute acid or dilute alkali, accounting for more than 80 percent of the total amount; and fourthly, the alcohol soluble protein is dissolved in 70 to 80 percent ethanol solution, and accounts for 1 to 5 percent of the total amount. Among them, albumin has the highest lysine content, globulin has high cystine content, and prolamin has low lysine and cystine contents. Although rice has a low protein content, rice protein is considered to have good nutritional quality because it is easily digested and absorbed and has a good flavor. Meanwhile, the rice protein has better quality than wheat protein and corn protein, contains high-quality lysine and has low irritability. According to the optimal amino acid composition standard published by the world trade organization, the amino acid composition mode of the rice protein is superior to that of casein and soy protein. Therefore, the rice protein has very wide application prospect in the food industry.
At present, the methods for producing rice protein peptide include acid-base method and enzymolysis method. Chinese patent application CN111217888A discloses a technology for extracting rice protein by an alkaline method, which comprises the steps of weighing rice flour which passes through a sieve in a fixed amount, preparing strong base with a certain concentration in a fixed amount for later use, dissolving the rice flour in a strong base solution according to a proper liquid-solid ratio, fully and uniformly mixing the rice flour at room temperature, leaching for a certain time to obtain a leaching solution, centrifuging the leaching solution under a high-speed centrifuge with proper power for separation, collecting supernatant, adjusting the pH of the supernatant to 4-6, carrying out acid precipitation, centrifuging after full deposition to obtain precipitate, washing with deionized water to remove albumin and residual inorganic salt for 3 times, and finally carrying out freeze drying to obtain the rice protein powder, wherein although the method is simple to operate and has high extraction rate, the defects of the alkaline extraction method are that the protein is severely denatured under the condition of high alkaline concentration, and meanwhile, the condensation reaction is generated between certain amino acids such as lysine and alanine or cystine under the alkaline environment, toxic substances are generated, the nutritive value of lysine is greatly reduced, and the finished product has dark color, bitter taste and poor practicability.
Chinese patent CN101695334B discloses a method for extracting rice protein from rice, which comprises the following steps: soaking, pulping, mixing, liquefying for one time, centrifuging, homogenizing, washing, centrifuging, performing enzymolysis, inactivating enzyme, sieving, washing, centrifuging, drying, pulverizing, and packaging to obtain rice protein. The method overcomes the damage of high-concentration alkali liquor to rice protein amino acid in the alkaline extraction process, but the purity and extraction rate of the extracted rice protein are low.
Therefore, it is a major technical problem to provide a simple and easy-to-operate preparation method which does not damage the nutrient substances of the rice protein peptide, and can effectively improve the extraction rate and solubility of the rice protein peptide, thereby promoting the digestibility of the rice protein peptide in the animal body.
Disclosure of Invention
In order to solve the problems in the prior art, the invention aims to provide a preparation method and application of rice protein peptide. The rice protein peptide prepared by the preparation method provided by the invention has high purity and solubility, is beneficial to the absorption of protein by animals, has an extraction rate as high as 85%, and effectively saves the cost.
The technical scheme of the invention is as follows:
a preparation method of rice protein peptide comprises the following steps:
s1, adding water into the rice, washing the rice, drying the rice, and storing the rice at the temperature of between 4 ℃ below zero and 10 ℃ below zero for 1 to 2 hours;
s2, extruding the rice obtained in the step S1 by using a screw extruder, crushing the extruded rice, sieving the crushed rice with a 60-80-mesh sieve, adding water with the volume being 10-20 times of that of the crushed rice, and homogenizing the crushed rice in a high-pressure homogenizer to obtain a homogenized liquid;
s3, adding the mixed solution into the homogeneous solution obtained in the step S2, standing for 1-3 hours, adjusting the pH to 4.0-6.0, heating to 35-50 ℃, adding the mixed enzyme twice, performing enzymolysis reaction, after the reaction is finished, heating to 80-90 ℃, performing enzyme inactivation for 10-15 min, cooling to 20-25 ℃, obtaining an enzymolysis solution, and filtering, concentrating and drying the enzymolysis solution to obtain the rice protein peptide.
In the step S2, the screw rotating speed of the screw extruder is 100-200 r/min, the extrusion temperature is 120-200 ℃, the extrusion pressure is 5-10 MPa, and the extrusion time is 20-40S.
The homogenizing time in the step S2 is 5-10 min, and the homogenizing pressure is 20-50 MPa.
The preparation method of the mixed solution in the step S3 includes: and (3) adding water which is 8-12 times of the total mass of tartaric acid, citric acid and dithiothreitol, and uniformly stirring to obtain a mixed solution.
The mass ratio of the tartaric acid to the citric acid to the dithiothreitol in the step S3 is 7: 1-5: 3 to 8.
The mass ratio of the tartaric acid to the citric acid to the dithiothreitol in the step S3 is 7: 3: 5.
the amount of the mixed enzyme added in the step S3 is 0.2-0.5% of the mass of the homogeneous liquid, and the enzymolysis time is 2-3 h.
The compound enzyme in the step S3 is composed of bromelain, papain and cellulase in a weight ratio of 7-11: 1-6: 2-5.
The compound enzyme in the step S3 is composed of bromelain, papain and cellulase according to the weight ratio of 9:4: 3.
In addition, the invention also provides application of the rice protein peptide in livestock feed.
In the prior art, although the enzymatic extraction can effectively solve the problem that strong base damages proteins during alkaline extraction, toxic substances generated by condensation among amino acids and bitter and astringent peculiar smell are avoided, the problems of low purity, low extraction rate, difficult absorption and digestion and the like of the extracted rice protein peptide still exist. The invention provides a preparation method of rice protein peptide, which comprises the steps of firstly, extruding the rice through a screw extruder after freezing preservation, enabling the rice to flow towards a die opening under the action of mechanical force in the screw extruder to generate high-temperature high-pressure high-shearing force and other actions, enabling the high-temperature rice to expand due to physical change when the high-temperature rice passes through the die opening because of negative pressure in a closed space, and physically cracking crude fiber to form a porous structure, enabling the rice to generate material texture recombination to a certain extent, and effectively improving the extraction rate and the solubility of the rice protein peptide when the rice protein peptide is extracted. And then the rice extruded by the screw is subjected to high-pressure homogenization treatment, so that the uniformity of the slurry can be further improved, the stretching of a protein molecular structure is facilitated, and the solubility of rice protein peptide is enhanced.
Furthermore, the mixed solution composed of tartaric acid, citric acid and dithiothreitol in a certain proportion added in the invention has synergistic effect, can promote the extraction of soluble peptide, effectively improve the extraction rate of rice protein peptide and enhance the water solubility of rice protein peptide. And then, after two times of enzymolysis reaction, by adding a complex enzyme consisting of bromelain, papain and cellulase, on one hand, the rice protein polypeptide chain can be hydrolyzed into a short peptide chain, so that the solubility of the rice protein peptide can be effectively improved, and on the other hand, non-protein substances in the rice raw material can be removed, so that the content and the purity of the protein are improved.
Compared with the prior art, the preparation method of the rice protein peptide provided by the invention has the following advantages:
(1) the preparation method of the rice protein peptide provided by the invention is simple, the operation is simple and convenient, the occurrence of bitter and astringent peculiar smell generated in the preparation of the rice protein peptide in the prior art can be solved through the synergistic effect of multiple steps, the extraction rate is up to 85%, and the preparation cost is effectively saved.
(2) The rice protein peptide prepared by the preparation method of the rice protein peptide provided by the invention has high purity which can reach more than 90%, and has good solubility which reaches more than 88%, thereby being very beneficial to the absorption of protein by animals.
Detailed Description
The present invention is further illustrated by the following description of specific embodiments, which are not intended to limit the invention, and various modifications and improvements can be made by those skilled in the art based on the basic idea of the invention, but the invention is within the protection scope of the invention.
The enzyme bromelain, CAS, item number 0511-00-7; papain, CAS:9001-73-4, cat #: 2662626, available from adadra biotechnology, guangzhou; cellulase, CAS:9012-54-8, purchased from Shenzhen Lefu Biotech, Inc.
The other reagents used in the invention are common reagents and can be purchased from conventional reagent production and sale companies.
Example 1A method for preparing rice protein peptide
The preparation method of the rice protein peptide comprises the following steps:
s1, adding water into rice, washing, drying, and storing at-10 deg.C for 1 h;
s2, extruding the rice obtained in the step S1 by adopting a screw extruder, crushing the extruded rice, sieving the crushed rice with a 60-mesh sieve, adding water with the volume being 10 times of that of the crushed rice, and homogenizing the crushed rice by using a high-pressure homogenizer to obtain a homogenized liquid; the screw rotating speed of the screw extruder is 100r/min, the extrusion temperature is 120 ℃, the extrusion pressure is 5MPa, and the extrusion time is 20 s; the homogenization time is 5min, and the homogenization pressure is 20 MPa;
s3, adding the mixed solution into the homogenized solution obtained in the step S2, standing for 1 hour, adjusting the pH to 4.0, heating to 35 ℃, adding the mixed enzyme twice, and performing enzymolysis reaction, wherein the amount of the mixed enzyme added each time is 0.2 percent of the mass of the homogenized solution, and the enzymolysis time is 2 hours each time; after the reaction is finished, heating to 80 ℃ for enzyme inactivation for 10min, cooling to 20 ℃ to obtain an enzymolysis solution, and filtering, concentrating and drying the enzymolysis solution to obtain the rice protein peptide.
The preparation method of the mixed solution in the step S3 includes: adding water with the weight 8 times of that of tartaric acid, citric acid and dithiothreitol, and uniformly stirring to obtain a mixed solution.
The mass ratio of the tartaric acid to the citric acid to the dithiothreitol in the step S3 is 7: 5: 8.
the compound enzyme in the step S3 in the step S3 is composed of bromelain, papain and cellulase according to the weight ratio of 7:6: 5.
Example 2A method for preparing rice protein peptide
The preparation method of the rice protein peptide comprises the following steps:
s1, adding water into rice, washing, drying, and storing at-7 ℃ for 1.5 h;
s2, extruding the rice obtained in the step S1 by adopting a screw extruder, crushing the extruded rice, sieving the crushed rice with a 70-mesh sieve, adding water with the volume 15 times that of the crushed rice, and homogenizing the crushed rice in a high-pressure homogenizer to obtain a homogenized liquid; the screw rotating speed of the screw extruder is 160r/min, the extrusion temperature is 170 ℃, the extrusion pressure is 8MPa, and the extrusion time is 30 s; the homogenization time is 8min, and the homogenization pressure is 40 MPa;
s3, adding the mixed solution into the homogenized solution obtained in the step S2, standing for 2 hours, adjusting the pH value to 5.0, heating to 45 ℃, adding the mixed enzyme twice, and performing enzymolysis reaction, wherein the amount of the mixed enzyme added each time is 0.35 percent of the mass of the homogenized solution, and the enzymolysis time is 2.5 hours each time; after the reaction is finished, heating to 85 ℃ for enzyme inactivation for 12min, cooling to 23 ℃ to obtain an enzymolysis solution, and filtering, concentrating and drying the enzymolysis solution to obtain the rice protein peptide.
The preparation method of the mixed solution in the step S3 includes: adding water with the weight 10 times of that of tartaric acid, citric acid and dithiothreitol, and uniformly stirring to obtain a mixed solution.
The mass ratio of the tartaric acid to the citric acid to the dithiothreitol in the step S3 is 7: 3: 5.
the compound enzyme in the step S3 is composed of bromelain, papain and cellulase according to the weight ratio of 9:4: 3.
Example 3A method for preparing Rice protein peptide
The preparation method of the rice protein peptide comprises the following steps:
s1, adding water into rice, washing, drying, and storing at-4 deg.C for 2 h;
s2, extruding the rice obtained in the step S1 by adopting a screw extruder, crushing the extruded rice, sieving the crushed rice with a 80-mesh sieve, adding water with the volume being 20 times of that of the crushed rice, and homogenizing the crushed rice by using a high-pressure homogenizer to obtain a homogenized liquid; the screw rotating speed of the screw extruder is 200r/min, the extrusion temperature is 200 ℃, the extrusion pressure is 10MPa, and the extrusion time is 40 s; the homogenization time is 10min, and the homogenization pressure is 50 MPa;
s3, adding the mixed solution into the homogenized solution obtained in the step S2, standing for 3 hours, adjusting the pH value to 6.0, heating to 50 ℃, adding the mixed enzyme twice, and performing enzymolysis reaction, wherein the amount of the mixed enzyme added each time is 0.5 percent of the mass of the homogenized solution, and the enzymolysis time is 3 hours each time; after the reaction is finished, heating to 90 ℃ for enzyme inactivation for 15min, cooling to 25 ℃ to obtain an enzymolysis solution, and filtering, concentrating and drying the enzymolysis solution to obtain the rice protein peptide.
The preparation method of the mixed solution in the step S3 includes: adding water with the weight 12 times of that of tartaric acid, citric acid and dithiothreitol, and uniformly stirring to obtain a mixed solution.
The mass ratio of the tartaric acid to the citric acid to the dithiothreitol in the step S3 is 7: 1: 3.
the compound enzyme in the step S3 in the step S3 is composed of bromelain, papain and cellulase according to the weight ratio of 11:1: 2.
Comparative example 1 preparation method of rice protein peptide
The preparation method of the rice protein peptide comprises the following steps:
s1, adding water into the rice, washing the rice, and drying;
s2, extruding the rice obtained in the step S1 by adopting a screw extruder, crushing the extruded rice, sieving the crushed rice with a 70-mesh sieve, adding water with the volume 15 times that of the crushed rice, and homogenizing the crushed rice in a high-pressure homogenizer to obtain a homogenized liquid; the screw rotating speed of the screw extruder is 160r/min, the extrusion temperature is 170 ℃, the extrusion pressure is 8MPa, and the extrusion time is 30 s; the homogenization time is 8min, and the homogenization pressure is 40 MPa;
s3, adding the mixed solution into the homogenized solution obtained in the step S2, standing for 2 hours, adjusting the pH value to 5.0, heating to 45 ℃, adding the mixed enzyme twice, and performing enzymolysis reaction, wherein the amount of the mixed enzyme added each time is 0.35 percent of the mass of the homogenized solution, and the enzymolysis time is 2.5 hours each time; after the reaction is finished, heating to 85 ℃ for enzyme inactivation for 12min, cooling to 23 ℃ to obtain an enzymolysis solution, and filtering, concentrating and drying the enzymolysis solution to obtain the rice protein peptide.
The preparation method of the mixed solution in the step S3 includes: adding water with the weight 10 times of that of tartaric acid, citric acid and dithiothreitol, and uniformly stirring to obtain a mixed solution.
The mass ratio of the tartaric acid to the citric acid to the dithiothreitol in the step S3 is 7: 3: 5.
the compound enzyme in the step S3 is composed of bromelain, papain and cellulase according to the weight ratio of 9:4: 3.
Comparative example 1 is different from example 2 in that it is not stored at-7 ℃ for 1.5h in step S1, and other parameters and operation are the same as example 2.
Comparative example 2 preparation method of rice protein peptide
The preparation method of the rice protein peptide comprises the following steps:
s1, adding water into rice, washing, drying, and storing at-7 ℃ for 1.5 h;
s2, crushing the rice obtained in the step S1, sieving the crushed rice with a 70-mesh sieve, adding water with the volume 15 times that of the crushed rice, and homogenizing the mixture in a high-pressure homogenizer to obtain a homogenized liquid; the homogenization time is 8min, and the homogenization pressure is 40 MPa;
s3, adding the mixed solution into the homogenized solution obtained in the step S2, standing for 2 hours, adjusting the pH value to 5.0, heating to 45 ℃, adding the mixed enzyme twice, and performing enzymolysis reaction, wherein the amount of the mixed enzyme added each time is 0.35 percent of the mass of the homogenized solution, and the enzymolysis time is 2.5 hours each time; after the reaction is finished, heating to 85 ℃ for enzyme inactivation for 12min, cooling to 23 ℃ to obtain an enzymolysis solution, and filtering, concentrating and drying the enzymolysis solution to obtain the rice protein peptide.
The preparation method of the mixed solution in the step S3 includes: adding water with the weight 10 times of that of tartaric acid, citric acid and dithiothreitol, and uniformly stirring to obtain a mixed solution.
The mass ratio of the tartaric acid to the citric acid to the dithiothreitol in the step S3 is 7: 3: 5.
the compound enzyme in the step S3 is composed of bromelain, papain and cellulase according to the weight ratio of 9:4: 3.
The comparative example 2 is different from the example 2 in that the rice is not subjected to the extrusion step using the screw extruder in the step S2, and other parameters and operations are the same as those of the example 2.
Comparative example 3 preparation method of rice protein peptide
The preparation method of the rice protein peptide comprises the following steps:
s1, adding water into rice, washing, drying, and storing at-7 ℃ for 1.5 h;
s2, extruding the rice obtained in the step S1 by adopting a screw extruder, crushing the extruded rice, sieving the crushed rice by a 70-mesh sieve, and adding water with the volume 15 times that of the crushed rice to obtain rice mixed liquor; the screw rotating speed of the screw extruder is 160r/min, the extrusion temperature is 170 ℃, the extrusion pressure is 8MPa, and the extrusion time is 30 s;
s3, adding the mixed solution into the rice mixed solution obtained in the step S2, standing for 2 hours, adjusting the pH value to 5.0, heating to 45 ℃, adding the mixed enzyme twice to perform enzymolysis reaction, wherein the amount of the mixed enzyme added each time is 0.35 percent of the mass of the rice mixed solution, and the enzymolysis time is 2.5 hours each time; after the reaction is finished, heating to 85 ℃ for enzyme inactivation for 12min, cooling to 23 ℃ to obtain an enzymolysis solution, and filtering, concentrating and drying the enzymolysis solution to obtain the rice protein peptide.
The preparation method of the mixed solution in the step S3 includes: adding water with the weight 10 times of that of tartaric acid, citric acid and dithiothreitol, and uniformly stirring to obtain a mixed solution.
The mass ratio of the tartaric acid to the citric acid to the dithiothreitol in the step S3 is 7: 3: 5.
the compound enzyme in the step S3 is composed of bromelain, papain and cellulase according to the weight ratio of 9:4: 3.
The comparative example 3 is different from the example 2 in that the high pressure homogenization treatment is not performed in the step S2, and other parameters and operations are the same as those of the example 2.
Comparative example 4 preparation method of rice protein peptide
The preparation method of the rice protein peptide is similar to that of the example 2.
In comparison with example 2, comparative example 4 is different in that dithiothreitol is not added to the mixed solution of step S3, and other parameters and operations are the same as those of example 2.
Comparative example 5 preparation method of Rice protein peptide
The preparation method of the rice protein peptide is similar to that of the example 2.
The comparative example 5 is different from the example 2 in that citric acid is not added to the mixed solution of the step S3, and other parameters and operations are the same as those of the example 2.
Comparative example 6 preparation method of Rice protein peptide
The preparation method of the rice protein peptide is similar to that of the example 2.
Compared with example 2, comparative example 6 is different in that the mass ratio of tartaric acid, citric acid and dithiothreitol in the mixed solution of step S3 is 1:1:1, other parameters and operations were the same as in example 2.
Comparative example 7 preparation method of rice protein peptide
The preparation method of the rice protein peptide is similar to that of the example 2.
Compared with example 2, comparative example 7 is different in that no cellulase is added to the complex enzyme in step S3, and other parameters and operations are the same as those of example 2.
Comparative example 8 preparation method of Rice protein peptide
The preparation method of the rice protein peptide is similar to that of the example 2.
Compared with the example 2, the comparative example 8 is different in that the complex enzyme in the step S3 is composed of bromelain, papain and cellulase according to the weight ratio of 1:1:1, and other parameters and operation are the same as those of the example 2.
Test example I measurement of solubility of Rice protein peptide
1. Test materials: the rice protein peptides prepared in examples 1 to 3 and comparative examples 1 to 8.
2. The test method comprises the following steps: the rice protein peptides prepared in examples 1 to 3 and comparative examples 1 to 8 were dissolved in water to prepare a 1% protein solution, stirred at 600r/min for 30min, centrifuged to obtain a supernatant, the protein content of the supernatant was determined by Kjeldahl method, and the protein solubility was determined by the ratio of the protein content in the supernatant to the total protein content.
3. Test results
The test results are shown in table 1.
TABLE 1 Rice protein peptide solubility assay
| Group of | Solubility (%) |
| Example 1 | 98.14 |
| Example 2 | 99.35 |
| Example 3 | 98.91 |
| Comparative example 1 | 90.74 |
| Comparative example 2 | 52.39 |
| Comparative example 3 | 69.74 |
| Comparative example 4 | 75.26 |
| Comparative example 5 | 77.81 |
| Comparative example 6 | 82.14 |
| Comparative example 7 | 92.17 |
| Comparative example 8 | 94.15 |
As can be seen from Table 1: the rice protein peptides prepared by the methods of the embodiments 1 to 3 of the invention have the solubility of more than 98%, wherein the embodiment 2 has the best effect, and the solubility is 99.35%, which is the best embodiment of the invention. The solubility of the rice protein peptides prepared by the methods of comparative examples 1 to 8 was lower than that of the rice protein peptides prepared by examples 1 to 3. This shows that the rice protein peptide obtained by the steps of the preparation method of the rice protein peptide provided by the invention are mutually influenced and act synergistically has good solubility.
Test example II determination of purity and extraction yield of Rice protein peptide
1. Test materials: the rice protein peptides prepared in examples 1 to 3 and comparative examples 1 to 8.
2. The test method comprises the following steps: the purity and extraction rate of the rice protein peptides prepared in examples 1 to 3 and comparative examples 1 to 8 were calculated by the following formulas.
(1)
(2)
The protein is detected by adopting a Kjeldahl method provided in GB 5009.5-2016 (determination of protein in food safety national standard food) for determination.
3. And (3) test results:
the test results are shown in table 2.
Table 2: purity and extraction rate determination of rice protein peptide
| Group of | Purity (%) | Extraction ratio (%) |
| Example 1 | 90.25 | 85.25 |
| Example 2 | 92.35 | 86.18 |
| Example 3 | 91.33 | 85.99 |
| Comparative example 1 | 82.56 | 77.23 |
| Comparative example 2 | 80.62 | 52.37 |
| Comparative example 3 | 72.48 | 75.26 |
| Comparative example 4 | 72.35 | 45.26 |
| Comparative example 5 | 76.15 | 48.52 |
| Comparative example 6 | 75.28 | 50.23 |
| Comparative example 7 | 44.24 | 55.36 |
| Comparative example 8 | 48.18 | 56.92 |
As can be seen from Table 2, the rice protein peptides prepared in examples 1 to 3 of the present invention were high in purity and extraction yield. Among them, the rice protein peptide obtained by the method of example 2 has the highest degree of hydrolysis and the highest extraction rate, and is the best example of the present invention. The purity and extraction rate of the rice protein peptide prepared by the method of comparative examples 1 to 8 are obviously reduced compared with those of examples 1 to 3, which shows that the rice protein peptide prepared by the preparation method of the rice protein peptide provided by the invention has higher purity and extraction rate through mutual influence and synergistic effect of the steps.
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 (10)
1. A preparation method of rice protein peptide is characterized by comprising the following steps:
s1, adding water into the rice, washing the rice, drying the rice, and storing the rice at the temperature of between 4 ℃ below zero and 10 ℃ below zero for 1 to 2 hours;
s2, extruding the rice obtained in the step S1 by using a screw extruder, crushing the extruded rice, sieving the crushed rice with a 60-80-mesh sieve, adding water with the volume being 10-20 times of that of the crushed rice, and homogenizing the crushed rice in a high-pressure homogenizer to obtain a homogenized liquid;
s3, adding the mixed solution into the homogeneous solution obtained in the step S2, standing for 1-3 hours, adjusting the pH to 4.0-6.0, heating to 35-50 ℃, adding the mixed enzyme twice, performing enzymolysis reaction, after the reaction is finished, heating to 80-90 ℃, performing enzyme inactivation for 10-15 min, cooling to 20-25 ℃, obtaining an enzymolysis solution, and filtering, concentrating and drying the enzymolysis solution to obtain the rice protein peptide.
2. The method for preparing rice protein peptide according to claim 1, wherein the screw speed of the screw extruder in the step S2 is 100-200 r/min, the extrusion temperature is 120-200 ℃, the extrusion pressure is 5-10 MPa, and the extrusion time is 20-40S.
3. The method for preparing rice protein peptide as claimed in claim 1, wherein the homogenization time in step S2 is 5-10 min, and the homogenization pressure is 20-50 MPa.
4. The method of preparing rice protein peptide as claimed in claim 1, wherein the method of preparing the mixed solution in the step S3 comprises: and (3) adding water which is 8-12 times of the total mass of tartaric acid, citric acid and dithiothreitol, and uniformly stirring to obtain a mixed solution.
5. The method for preparing rice protein peptide as claimed in claim 4, wherein the mass ratio of tartaric acid, citric acid and dithiothreitol in step S3 is 7: 1-5: 3 to 8.
6. The method for preparing rice protein peptide as claimed in claim 5, wherein the mass ratio of tartaric acid, citric acid and dithiothreitol in step S3 is 7: 3: 5.
7. the method for preparing rice protein peptide as claimed in claim 1, wherein the amount of the mixed enzyme added in each step S3 is 0.2-0.5% of the mass of the homogenized solution, and the enzymolysis time is 2-3 h.
8. The method for preparing rice protein peptide as claimed in claim 1, wherein the complex enzyme in the step S3 is bromelain, papain and cellulase in a weight ratio of 7-11: 1-6: 2-5.
9. The method for preparing rice protein peptide as claimed in claim 8, wherein the complex enzyme in step S3 is bromelain, papain and cellulase in a weight ratio of 9:4: 3.
10. Use of a rice protein peptide according to any one of claims 1 to 9 in livestock feed.
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