CN111485010B - A kind of preparation method of cholesterol lowering protein - Google Patents
A kind of preparation method of cholesterol lowering protein Download PDFInfo
- Publication number
- CN111485010B CN111485010B CN202010408766.9A CN202010408766A CN111485010B CN 111485010 B CN111485010 B CN 111485010B CN 202010408766 A CN202010408766 A CN 202010408766A CN 111485010 B CN111485010 B CN 111485010B
- Authority
- CN
- China
- Prior art keywords
- protein
- cholesterol
- modification
- ultrasonic
- lowering
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 68
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 68
- 238000002360 preparation method Methods 0.000 title claims abstract description 8
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 title description 24
- 235000012000 cholesterol Nutrition 0.000 title description 12
- 230000000694 effects Effects 0.000 claims abstract description 49
- 230000029087 digestion Effects 0.000 claims abstract description 33
- 230000004048 modification Effects 0.000 claims abstract description 25
- 238000012986 modification Methods 0.000 claims abstract description 25
- 102000004190 Enzymes Human genes 0.000 claims abstract description 16
- 108090000790 Enzymes Proteins 0.000 claims abstract description 16
- 230000008878 coupling Effects 0.000 claims abstract description 14
- 238000010168 coupling process Methods 0.000 claims abstract description 14
- 238000005859 coupling reaction Methods 0.000 claims abstract description 14
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 claims abstract description 9
- 230000007071 enzymatic hydrolysis Effects 0.000 claims abstract description 7
- 230000007062 hydrolysis Effects 0.000 claims abstract description 5
- 238000006460 hydrolysis reaction Methods 0.000 claims abstract description 5
- 238000001816 cooling Methods 0.000 claims abstract description 3
- 230000002255 enzymatic effect Effects 0.000 claims abstract description 3
- 239000012460 protein solution Substances 0.000 claims abstract description 3
- 239000000725 suspension Substances 0.000 claims abstract description 3
- 235000018102 proteins Nutrition 0.000 claims description 59
- 238000000338 in vitro Methods 0.000 claims description 28
- 230000002496 gastric effect Effects 0.000 claims description 15
- 230000009145 protein modification Effects 0.000 claims description 13
- 235000007164 Oryza sativa Nutrition 0.000 claims description 9
- 238000000034 method Methods 0.000 claims description 9
- 235000009566 rice Nutrition 0.000 claims description 9
- 108091005804 Peptidases Proteins 0.000 claims description 5
- 239000004365 Protease Substances 0.000 claims description 5
- 102000014171 Milk Proteins Human genes 0.000 claims description 3
- 108010011756 Milk Proteins Proteins 0.000 claims description 3
- 235000021239 milk protein Nutrition 0.000 claims description 3
- 230000008569 process Effects 0.000 claims description 2
- 238000012545 processing Methods 0.000 claims description 2
- 238000009210 therapy by ultrasound Methods 0.000 claims description 2
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims 2
- 240000007594 Oryza sativa Species 0.000 claims 1
- 102000035118 modified proteins Human genes 0.000 abstract description 31
- 108091005573 modified proteins Proteins 0.000 abstract description 31
- 238000002604 ultrasonography Methods 0.000 abstract description 12
- 108090000145 Bacillolysin Proteins 0.000 abstract description 10
- 102000035092 Neutral proteases Human genes 0.000 abstract description 10
- 108091005507 Neutral proteases Proteins 0.000 abstract description 10
- 230000009144 enzymatic modification Effects 0.000 abstract description 3
- 239000002994 raw material Substances 0.000 abstract description 2
- 239000000413 hydrolysate Substances 0.000 abstract 1
- 230000000968 intestinal effect Effects 0.000 abstract 1
- 239000000047 product Substances 0.000 abstract 1
- 238000001694 spray drying Methods 0.000 description 25
- 239000000203 mixture Substances 0.000 description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 19
- 210000001035 gastrointestinal tract Anatomy 0.000 description 18
- 229940088598 enzyme Drugs 0.000 description 14
- 238000004088 simulation Methods 0.000 description 13
- 230000000052 comparative effect Effects 0.000 description 12
- 235000013336 milk Nutrition 0.000 description 12
- 239000008267 milk Substances 0.000 description 12
- 210000004080 milk Anatomy 0.000 description 12
- 238000005303 weighing Methods 0.000 description 12
- 238000006243 chemical reaction Methods 0.000 description 9
- 239000000463 material Substances 0.000 description 9
- 241000209094 Oryza Species 0.000 description 8
- 230000009849 deactivation Effects 0.000 description 8
- 239000012153 distilled water Substances 0.000 description 6
- 238000003756 stirring Methods 0.000 description 6
- 230000035790 physiological processes and functions Effects 0.000 description 5
- 102000035195 Peptidases Human genes 0.000 description 3
- 230000000415 inactivating effect Effects 0.000 description 3
- 238000002525 ultrasonication Methods 0.000 description 3
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 210000000936 intestine Anatomy 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 210000002784 stomach Anatomy 0.000 description 2
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 235000019750 Crude protein Nutrition 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 208000035150 Hypercholesterolemia Diseases 0.000 description 1
- 108060003951 Immunoglobulin Proteins 0.000 description 1
- 108010019160 Pancreatin Proteins 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- 102000007079 Peptide Fragments Human genes 0.000 description 1
- 108010033276 Peptide Fragments Proteins 0.000 description 1
- 230000010757 Reduction Activity Effects 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 229920002494 Zein Polymers 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 208000026106 cerebrovascular disease Diseases 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 210000004351 coronary vessel Anatomy 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 230000005714 functional activity Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 102000018358 immunoglobulin Human genes 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000000693 micelle Substances 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 238000000053 physical method Methods 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000007065 protein hydrolysis Effects 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 125000003396 thiol group Chemical class [H]S* 0.000 description 1
- 238000004506 ultrasonic cleaning Methods 0.000 description 1
- 239000005019 zein Substances 0.000 description 1
- 229940093612 zein Drugs 0.000 description 1
Classifications
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/06—Antihyperlipidemics
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Molecular Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Pharmacology & Pharmacy (AREA)
- Hematology (AREA)
- Veterinary Medicine (AREA)
- Analytical Chemistry (AREA)
- Diabetes (AREA)
- Biophysics (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Obesity (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Engineering & Computer Science (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
本发明公开了一种降胆固醇蛋白质的制备方法，属于功能蛋白制备领域。先将蛋白溶液或者悬浊液预热后，采用高强度超声修饰蛋白；然后对蛋白进行低强度超声—酶解耦合改性，超声修饰后的蛋白加入中性蛋白酶进行酶解改性，以胃肠消化后产物的最高降胆固醇活性为水解终点，停止酶解，灭酶，冷却后进行浓缩和喷雾干燥，制备成降胆固醇功能蛋白。在此条件下，酶解物的降胆固醇活性和原料相比提高了4倍以上，和高强度超声修饰蛋白相比提高了2倍以上，和低强度超声—酶解耦合改性蛋白相比增加了44.90％以上。The invention discloses a preparation method of a cholesterol-lowering protein, and belongs to the field of functional protein preparation. After preheating the protein solution or suspension, high-intensity ultrasound is used to modify the protein; then the protein is modified by low-intensity ultrasound-enzymatic hydrolysis coupling, and the protein after ultrasonic modification is added with neutral protease for enzymatic modification. The highest cholesterol-lowering activity of the product after intestinal digestion is the end point of hydrolysis, the enzymatic hydrolysis is stopped, the enzyme is inactivated, concentrated and spray-dried after cooling to prepare a cholesterol-lowering functional protein. Under these conditions, the cholesterol-lowering activity of the enzymatic hydrolysate was increased by more than 4 times compared with the raw material, increased by more than 2 times compared with the high-intensity ultrasound-modified protein, and increased compared with the low-intensity ultrasound-enzyme hydrolysis coupled modified protein. over 44.90%.
Description
Technical Field
The invention belongs to the field of functional protein preparation, and particularly relates to a method for preparing functional protein for reducing cholesterol through protein modification.
Background
Food-borne proteins in the nature except a few proteins such as immunoglobulin can resist the digestion of the stomach and intestine, and further play a physiological role in improving the immunity and the like, and most of the proteins are degraded into small molecular peptides and amino acids through the stomach and intestine digestion to play a nutritional function. In short, after most of proteins are digested by gastrointestinal tract, the high-level structure of the proteins is damaged, and the physiological function value cannot be embodied. If the protein is modified by adopting a proper treatment method, some beneficial high-level structures can be reserved before complete digestion after the protein enters the gastrointestinal tract, and partial physiological functions can be realized. The enzyme method modified protein can carry out enzymolysis on long-chain protein into short-chain protein, and the short-chain protein can theoretically generate peptide fragments with smaller molecular weight and physiological functions under the action of gastrointestinal digestion, so as to further exert the physiological functions.
The physiological function of a protein depends on the higher order structure of the protein and, in addition to enzymatic modification of the protein, other physical methods can modify the protein, for example, ultrasonication can change the conformation of the protein, ultrasonication can increase the surface hydrophobicity, the enzymatic hydrolysis, the solubility, the thiol content of the protein, and change the composition of the secondary structure of the protein (Yang et al, Effects of low power condensation multi-stage chromatography on the enzymatic and structural characterization of degraded protein 2017; Yang et al, Effects and mechanism of adsorbed prediction of protein on the metallic reaction of protein-hydrolysis protein, 2020), ultrasonication can modify the protein by changing the conformation of the protein, and further improve the functional activity of the functional protein.
Hypercholesterolemia refers to the increase of blood fat due to the increase of cholesterol in blood beyond the normal range, which leads to the formation of arterial plaque in coronary artery wall, thus causing atherosclerosis and increasing the risk of cardiovascular and cerebrovascular diseases. At present, researches on the function of reducing cholesterol by using ultrasonic modification and combining with an enzymatic modification technology to prepare functional protein after the functional protein is digested by gastrointestinal protease in a digestive tract are not reported.
Disclosure of Invention
The invention aims to provide a preparation method of protein for reducing cholesterol, which is to adopt ultrasonic-enzymolysis coupling modification on the basis of ultrasonic modification to further obtain functional protein for reducing cholesterol activity.
In order to achieve the above purpose, the specific technical solution is as follows: a preparation method of protein for reducing cholesterol comprises the following steps:
(1) high-intensity ultrasonic modification of protein: preheating a protein solution or suspension (the concentration is 10-100 g/L), and modifying by using ultrasound, wherein the ultrasound processing parameters are as follows: the ultrasonic power density is 50-500W/L, and the ultrasonic treatment time is 5-60 min.
(2) Low-intensity ultrasonic-enzymolysis coupling modification of protein: adding protease into the ultrasonically modified protein for enzymolysis modification, then adjusting the temperature to 50 ℃, adjusting the pH value to 7.0, adding 5% neutral protease (E/S) for enzymolysis, keeping the enzymolysis condition constant, performing the enzymolysis modification process in a low-intensity ultrasonic field, controlling the ultrasonic power density to be 5-20W/L, taking the highest cholesterol reduction activity of a product obtained after in vitro simulation of gastrointestinal tract digestion as a hydrolysis end point, stopping enzymolysis, inactivating enzyme, cooling, concentrating, and performing spray drying to prepare the functional protein for reducing cholesterol.
Wherein the protein in the step (1) is water-soluble protein, such as milk protein and egg white protein. It may also be a protein that is not readily soluble in water, such as zein, rice protein.
The invention has the advantages that: the invention adopts high-intensity ultrasound to modify the protein structure, then adopts low-intensity ultrasound-enzymolysis coupling to modify the protein, and takes the highest cholesterol-reducing activity of the zymolyte after the digestion of the gastrointestinal tract is simulated in vitro as the judgment basis of the enzymolysis end point. Under the condition, the cholesterol-reducing activity of the zymolyte is greatly increased, compared with the raw material, the cholesterol-reducing activity is improved by more than 4 times, compared with the high-intensity ultrasonic modified protein, the cholesterol-reducing activity is improved by more than 2 times, and compared with the low-intensity ultrasonic-enzymolysis coupling modified protein, the cholesterol-reducing activity is increased by more than 44.90%.
The technical solution of the present invention is further described in detail by the following examples.
Detailed Description
Terms used in the present invention have generally meanings as commonly understood by one of ordinary skill in the art, unless otherwise specified. The present invention is described in further detail below with reference to specific examples and with reference to the data. It is to be understood that these examples are for illustrative purposes only and are not to be construed as limiting the scope of the invention, which is intended to be covered by the appended claims.
In the following examples, various procedures and methods not described in detail are conventional methods well known in the art. The source, trade name and composition of the reagents used are indicated at the first appearance and the same reagents used thereafter are the same as indicated for the first time unless otherwise specified.
Proteases of the present example and the comparative example were purchased from Sigma.
The water-soluble proteins of the present example and the comparative example, taking milk protein as an example, are all bright best pasteurized fresh milk (protein content 3.4%, fat content 0%).
The water-insoluble proteins of the present example and the comparative example, taking rice protein as an example, were provided by biological technologies of Beijing Yangshan, lake (crude protein content 57.90%, starch content 11.73%).
The degree of hydrolysis in the examples and the comparative examples was measured by the pH-stat method.
The ultrasonic equipment of the present example and the comparative example was a KQ-300DE type numerical control ultrasonic cleaning apparatus, which was purchased from kunshan ultrasonic instruments ltd.
The following methods were used for simulating gastrointestinal digestion in vitro in both the present example and the control example: adding pepsin into the enzymolysis product according to the enzyme adding amount of 2% (E/S), carrying out enzymolysis for 2 hours, inactivating enzyme after the enzymolysis is finished, adding pancreatin according to the enzyme adding amount of 4% (E/S), carrying out enzymolysis for 4 hours, inactivating enzyme after the enzymolysis is finished, centrifuging, and measuring the cholesterol-reducing activity.
Measurement of Cholesterol lowering Activity in the present example and comparative example refer to the measurement method of the inhibition rate of cholesterol micelle solubility (Nagaoka et al, Biochemical and Biophysical Research Communications,2001,281: 11-17).
Comparative example 1
Weighing 3L of milk, adjusting the temperature to 50 deg.C, preheating, and simulating gastrointestinal digestion in vitro to obtain cholesterol-lowering activity of 7.24%. After concentration and spray drying, the mixture is added with water for redissolution (keeping the protein content of the mixture unchanged), the cholesterol-reducing activity of the mixture after in vitro simulation of gastrointestinal tract digestion is 0.43 percent, and the activity of the mixture after spray drying is kept 5.94 percent.
Comparative example 2
Weighing 3L of milk, adjusting the temperature of the milk to 50 ℃, preheating, and carrying out protein modification by adopting high-intensity ultrasound, wherein the modification time is 5min, and the power density is 50W/L. The cholesterol-lowering activity of the modified protein reaches 11.35 percent after the modified protein is subjected to in vitro gastrointestinal tract digestion simulation, and the modified protein product is obtained after concentration and spray drying. After being reconstituted by adding water (keeping the protein content unchanged), the cholesterol-lowering activity of the composition after in vitro simulated gastrointestinal digestion is measured to be 0.90%, and the activity after spray drying is kept to be 7.93%.
Comparative example 3
Weighing 3L of milk, adjusting the temperature of the milk to 50 ℃, preheating, adjusting the pH to 7.0, adding 5% E/S neutral protease to perform low-intensity ultrasonic-enzymolysis coupling modification on the protein, and maintaining the temperature and the pH constant, wherein the ultrasonic power density is 5W/L. When the cholesterol-reducing activity of the modified protein reaches the highest value of 28.73 percent after the modified protein is subjected to in vitro gastrointestinal tract digestion simulation, the enzymolysis reaction is stopped, and the modified protein product is obtained after enzyme deactivation, concentration and spray drying. After being reconstituted by adding water (keeping the protein content unchanged), the cholesterol-lowering activity of the composition after in vitro simulated gastrointestinal digestion is measured to be 6.24%, and the activity after spray drying is kept to be 21.72%.
Example 1
Weighing 3L of milk, adjusting the temperature of the milk to 50 ℃, preheating, and carrying out protein modification by adopting high-intensity ultrasound, wherein the modification time is 5min, and the power density is 50W/L; after the temperature is adjusted to 50 ℃, the pH value is 7.0, 5 percent of E/S neutral protease is added to carry out low-intensity ultrasonic-enzymolysis coupling modification on the protein, the temperature and the pH value are kept constant, and the ultrasonic power density is 5W/L. When the cholesterol-lowering activity of the modified protein reaches 41.63% of the highest value after in vitro gastrointestinal tract digestion simulation, the enzymolysis reaction is stopped, and the modified protein product is obtained after enzyme deactivation, concentration and spray drying. After being reconstituted with water (keeping the protein content unchanged), the cholesterol-lowering activity of the composition after in vitro simulated gastrointestinal digestion is determined to be 21.56%, and the activity after spray drying is kept to be 51.79%.
Example 2
Weighing 3L of milk, adjusting the temperature of the milk to 50 ℃, preheating, and performing protein modification by adopting high-intensity ultrasound, wherein the modification time is 20min and the power density is 200W/L; after the temperature is adjusted to 50 ℃, the pH value is 7.0, 5 percent of E/S neutral protease is added to carry out low-intensity ultrasonic-enzymolysis coupling modification on the protein, the temperature and the pH value are kept constant, and the ultrasonic power density is 10W/L. When the cholesterol-lowering activity of the modified protein reaches 53.28% of the highest value after in vitro gastrointestinal tract digestion simulation, the enzymolysis reaction is stopped, and the modified protein product is obtained after enzyme deactivation, concentration and spray drying. After being reconstituted with water (keeping the protein content unchanged), the cholesterol-lowering activity of the composition after in vitro simulated gastrointestinal digestion is determined to be 32.43%, and the activity after spray drying is maintained at 60.87%.
Example 3
Weighing 3L of milk, adjusting the temperature of the milk to 50 ℃, preheating, and performing protein modification by adopting high-intensity ultrasound, wherein the modification time is 60min, and the power density is 500W/L; after the temperature is adjusted to 50 ℃, the pH value is 7.0, 5 percent of E/S neutral protease is added to carry out low-intensity ultrasonic-enzymolysis coupling modification on the protein, the temperature and the pH value are kept constant, and the ultrasonic power density is 20W/L. When the cholesterol-reducing activity of the modified protein reaches the highest value of 48.29 percent after the modified protein is subjected to in vitro gastrointestinal tract digestion simulation, the enzymolysis reaction is stopped, and the modified protein product is obtained after enzyme deactivation, concentration and spray drying. After being reconstituted with water (keeping the protein content constant), the cholesterol-lowering activity of the composition after in vitro simulated gastrointestinal digestion was determined to be 26.72%, and the activity after spray drying was determined to be 55.33%.
Comparative example 4
Weighing 10g of rice protein, adding 1L of distilled water to prepare a material with the concentration of 10g/L, uniformly stirring, adjusting the temperature to 50 ℃ for preheating, and simulating gastrointestinal tract digestion in vitro to obtain the cholesterol-reducing activity of 3.37%. After concentration and spray drying, the mixture is added with water for redissolution (keeping the protein content of the mixture unchanged), the cholesterol-reducing activity of the mixture after in vitro simulation of gastrointestinal tract digestion is 0.21 percent, and the activity of the mixture after spray drying is kept 6.24 percent.
Comparative example 5
Weighing 10g of rice protein, adding 1L of distilled water to prepare a material with the concentration of 10g/L, uniformly stirring, adjusting the temperature to 50 ℃ for preheating, and performing protein modification by adopting high-intensity ultrasound for 5min with the power density of 50W/L. The cholesterol-lowering activity of the modified protein reaches 6.17 percent after the modified protein is subjected to in vitro gastrointestinal tract digestion simulation, and a modified protein product is obtained after concentration and spray drying. After being reconstituted with water (keeping the protein content unchanged), the cholesterol-lowering activity of the composition after in vitro simulated gastrointestinal digestion is determined to be 0.54%, and the activity after spray drying is kept to be 8.75%.
Comparative example 6
Weighing 10g of rice protein, adding 1L of distilled water to prepare a material with the concentration of 10g/L, uniformly stirring, adjusting the temperature to 50 ℃ for preheating, adjusting the pH to 7.0, adding 5% E/S neutral protease to perform low-intensity ultrasonic-enzymolysis coupling modification on the protein, and maintaining the temperature and the pH constant, wherein the ultrasonic power density is 5W/L. When the cholesterol-reducing activity of the modified protein reaches the highest value of 14.56 percent after the modified protein is subjected to in vitro gastrointestinal tract digestion simulation, the enzymolysis reaction is stopped, and the modified protein product is obtained after enzyme deactivation, concentration and spray drying. After being reconstituted by adding water (keeping the protein content unchanged), the cholesterol-lowering activity of the composition after in vitro simulated gastrointestinal digestion is measured to be 3.38%, and the activity after spray drying is kept to be 23.21%.
Example 4
Weighing 10g of rice protein, adding 1L of distilled water to prepare a material with the concentration of 10g/L, uniformly stirring, adjusting the temperature of the material to 50 ℃ for preheating, and performing protein modification by adopting high-intensity ultrasound, wherein the modification time is 5min and the power density is 50W/L; after the temperature is adjusted to 50 ℃, the pH value is 7.0, 5 percent of E/S neutral protease is added to carry out low-intensity ultrasonic-enzymolysis coupling modification on the protein, the temperature and the pH value are kept constant, and the ultrasonic power density is 5W/L. When the cholesterol-reducing activity of the modified protein reaches the highest value of 26.35 percent after the modified protein is subjected to in vitro gastrointestinal tract digestion simulation, the enzymolysis reaction is stopped, and the modified protein product is obtained after enzyme deactivation, concentration and spray drying. After being reconstituted with water (keeping the protein content unchanged), the cholesterol-lowering activity of the composition after in vitro simulated gastrointestinal digestion is measured to be 11.63%, and the activity after spray drying is kept to be 44.13%.
Example 5
Weighing 50g of rice protein, adding 1L of distilled water to prepare a material with the concentration of 50g/L, uniformly stirring, adjusting the temperature of the material to 50 ℃ for preheating, and performing protein modification by adopting high-intensity ultrasound, wherein the modification time is 20min and the power density is 200W/L; after the temperature is adjusted to 50 ℃, the pH value is 7.0, 5 percent of E/S neutral protease is added to carry out low-intensity ultrasonic-enzymolysis coupling modification on the protein, the temperature and the pH value are kept constant, and the ultrasonic power density is 10W/L. When the cholesterol-lowering activity of the modified protein reaches 43.20% of the highest value after in vitro gastrointestinal tract digestion simulation, the enzymolysis reaction is stopped, and the modified protein product is obtained after enzyme deactivation, concentration and spray drying. After being reconstituted with water (keeping the protein content unchanged), the cholesterol-lowering activity of the composition after in vitro simulated gastrointestinal digestion is measured to be 25.42%, and the activity after spray drying is kept to be 58.84%.
Example 6
Weighing 100g of rice protein, adding 1L of distilled water to prepare a material with the concentration of 100g/L, uniformly stirring, adjusting the temperature of the material to 50 ℃ for preheating, and performing protein modification by adopting high-intensity ultrasound, wherein the modification time is 60min and the power density is 500W/L; after the temperature is adjusted to 50 ℃, the pH value is 7.0, 5 percent of E/S neutral protease is added to carry out low-intensity ultrasonic-enzymolysis coupling modification on the protein, the temperature and the pH value are kept constant, and the ultrasonic power density is 20W/L. When the cholesterol-lowering activity of the modified protein reaches 54.13% of the highest value after in vitro gastrointestinal tract digestion simulation, the enzymolysis reaction is stopped, and the modified protein product is obtained after enzyme deactivation, concentration and spray drying. After being reconstituted with water (keeping the protein content unchanged), the cholesterol-lowering activity of the composition after in vitro simulated gastrointestinal digestion is determined to be 27.94%, and the activity after spray drying is kept to be 51.62%.
Claims (1)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010408766.9A CN111485010B (en) | 2020-05-14 | 2020-05-14 | A kind of preparation method of cholesterol lowering protein |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010408766.9A CN111485010B (en) | 2020-05-14 | 2020-05-14 | A kind of preparation method of cholesterol lowering protein |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111485010A CN111485010A (en) | 2020-08-04 |
| CN111485010B true CN111485010B (en) | 2022-03-01 |
Family
ID=71790429
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010408766.9A Active CN111485010B (en) | 2020-05-14 | 2020-05-14 | A kind of preparation method of cholesterol lowering protein |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111485010B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116391787A (en) * | 2023-05-05 | 2023-07-07 | 安徽农业大学 | Method for preparing egg white protein cholesterol-reducing peptide through enzymolysis modification |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP3523248B2 (en) * | 2003-02-21 | 2004-04-26 | 明治乳業株式会社 | Cholesterol-reducing peptide |
| CN102864201A (en) * | 2012-10-09 | 2013-01-09 | 陕西同正科技有限公司 | Preparation method of corn active peptides |
| CN103695518B (en) * | 2013-12-30 | 2016-04-06 | 东北农业大学 | Whey protein peptide is utilized to reduce product and the preparation method of serum cholesterol level |
| CN105886588A (en) * | 2016-04-28 | 2016-08-24 | 李健 | Preparation method of bovine casein cholesterol reduction peptide |
| CN106538821B (en) * | 2016-11-08 | 2020-08-18 | 大连工业大学 | Functional protein with bile acid adsorption capacity and preparation method thereof |
| CN109628536A (en) * | 2018-11-27 | 2019-04-16 | 安徽博悦生物科技有限公司 | A kind of preparation method of Soyprotein peptide |
-
2020
- 2020-05-14 CN CN202010408766.9A patent/CN111485010B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN111485010A (en) | 2020-08-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101861909B (en) | Method for modification of rice protein and oryzenin with protein glutaminase | |
| JPH0665280B2 (en) | Protein gelling agent and protein gelling method using the same | |
| CN101427728B (en) | A method for improving the performance of wheat germ protein by using ultrasonic waves | |
| CN111631261A (en) | A kind of liquid milk preparation method based on reducing milk allergy | |
| CN113080468B (en) | Slow-digestion casein nano micelle powder and preparation method thereof | |
| CN111363772A (en) | Method for preparing collagen peptide by hydrolyzing bovine bone and collagen peptide thereof | |
| CN109123067A (en) | A kind of method reducing cow's milk protein sensitization and low sensitization cow's milk protein | |
| CN111485010B (en) | A kind of preparation method of cholesterol lowering protein | |
| CN103215333A (en) | Method for preparing angiotensin-I-converting enzyme (ACE) inhibitory peptide by using enzymolysis goat milk casein | |
| CN111990639A (en) | A method for preparing SPH-EGCG complex by utilizing polyphenol precipitation hydrolyzed peptide | |
| CN109897879A (en) | A kind of preparation method of subsensitivety ovalbumin peptide | |
| CN103444979A (en) | Method for lowering albumen allergenicity by immobilized enzyme hydrolysis | |
| CN102823715B (en) | A method for preparing biologically active peptides by a fully biomimetic digestion model method | |
| CN117363681A (en) | Bonito elastin peptide and preparation method thereof | |
| JP2023530376A (en) | Method for producing collagen peptide | |
| CN104131055A (en) | Preparation method for phycoerythrin ACE inhibitory peptide | |
| WO2023082523A1 (en) | Method for improving extraction rate of chondroitin sulfate prepared from tilapia skull | |
| CN109880874A (en) | A kind of preparation method of rice active peptide, rice active peptide and its application | |
| JP2572716B2 (en) | Novel transglutaminase | |
| JP2003250460A (en) | Method for modifying functionality of milk protein | |
| CN1330771C (en) | Method of preparing peptide having trypsin inhibition activity by using complex enzyme enzymolysis oyster | |
| CN118638175A (en) | A plant protein isolate and preparation method thereof | |
| JPS58158137A (en) | Method for producing protein for hydrolysis | |
| RU2529707C2 (en) | Method of producing whey protein hydrolysate with high degree of hydrolysis and low residual antigenicity | |
| JP2502531B2 (en) | Modified protein manufacturing method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20220127 Address after: 067000 Anyuan Road, Shuangqiao District, Chengde City, Hebei Province Applicant after: CHENGDE MEDICAL University Applicant after: jiangsu university Address before: 067000 Anyuan Road, Shuangqiao District, Chengde City, Hebei Province Applicant before: CHENGDE MEDICAL University |
|
| TA01 | Transfer of patent application right | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |