CN111358009A - Plant peptide and preparation method and application thereof - Google Patents
Plant peptide and preparation method and application thereof Download PDFInfo
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- CN111358009A CN111358009A CN202010183147.4A CN202010183147A CN111358009A CN 111358009 A CN111358009 A CN 111358009A CN 202010183147 A CN202010183147 A CN 202010183147A CN 111358009 A CN111358009 A CN 111358009A
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- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 98
- 238000002360 preparation method Methods 0.000 title description 16
- 244000068988 Glycine max Species 0.000 claims abstract description 55
- 235000010469 Glycine max Nutrition 0.000 claims abstract description 55
- 108010038807 Oligopeptides Proteins 0.000 claims abstract description 55
- 102000015636 Oligopeptides Human genes 0.000 claims abstract description 55
- 241000196324 Embryophyta Species 0.000 claims abstract description 45
- 235000013373 food additive Nutrition 0.000 claims abstract description 32
- 239000002778 food additive Substances 0.000 claims abstract description 32
- 240000008042 Zea mays Species 0.000 claims abstract description 30
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims abstract description 30
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims abstract description 30
- 235000005822 corn Nutrition 0.000 claims abstract description 30
- 239000002994 raw material Substances 0.000 claims abstract description 30
- 239000005715 Fructose Substances 0.000 claims abstract description 22
- 229930091371 Fructose Natural products 0.000 claims abstract description 22
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 claims abstract description 22
- 238000006243 chemical reaction Methods 0.000 claims description 33
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- 108010007119 flavourzyme Proteins 0.000 claims description 19
- 238000004108 freeze drying Methods 0.000 claims description 19
- 239000002131 composite material Substances 0.000 claims description 16
- 210000004369 blood Anatomy 0.000 claims description 15
- 239000008280 blood Substances 0.000 claims description 15
- 230000000415 inactivating effect Effects 0.000 claims description 15
- 238000000108 ultra-filtration Methods 0.000 claims description 15
- 235000019658 bitter taste Nutrition 0.000 claims description 12
- 235000005976 Citrus sinensis Nutrition 0.000 claims description 10
- 240000002319 Citrus sinensis Species 0.000 claims description 10
- UEDUENGHJMELGK-HYDKPPNVSA-N Stevioside Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@]12C(=C)C[C@@]3(C1)CC[C@@H]1[C@@](C)(CCC[C@]1([C@@H]3CC2)C)C(=O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O UEDUENGHJMELGK-HYDKPPNVSA-N 0.000 claims description 10
- 229930189775 mogroside Natural products 0.000 claims description 10
- OHHNJQXIOPOJSC-UHFFFAOYSA-N stevioside Natural products CC1(CCCC2(C)C3(C)CCC4(CC3(CCC12C)CC4=C)OC5OC(CO)C(O)C(O)C5OC6OC(CO)C(O)C(O)C6O)C(=O)OC7OC(CO)C(O)C(O)C7O OHHNJQXIOPOJSC-UHFFFAOYSA-N 0.000 claims description 10
- 229940013618 stevioside Drugs 0.000 claims description 10
- 235000019202 steviosides Nutrition 0.000 claims description 10
- 108091005658 Basic proteases Proteins 0.000 claims description 9
- 108090000145 Bacillolysin Proteins 0.000 claims description 8
- 108091005507 Neutral proteases Proteins 0.000 claims description 8
- 102000035092 Neutral proteases Human genes 0.000 claims description 8
- 239000012528 membrane Substances 0.000 claims description 8
- 239000000706 filtrate Substances 0.000 claims description 7
- 238000000034 method Methods 0.000 claims description 7
- 238000004519 manufacturing process Methods 0.000 claims description 6
- 239000006228 supernatant Substances 0.000 claims description 6
- 108010027597 alpha-chymotrypsin Proteins 0.000 claims description 3
- 210000004185 liver Anatomy 0.000 claims description 3
- 238000005119 centrifugation Methods 0.000 claims description 2
- 238000001816 cooling Methods 0.000 claims description 2
- 239000003814 drug Substances 0.000 claims description 2
- 235000013376 functional food Nutrition 0.000 claims description 2
- 206010019133 Hangover Diseases 0.000 claims 1
- 230000009849 deactivation Effects 0.000 claims 1
- 235000013402 health food Nutrition 0.000 claims 1
- 239000000243 solution Substances 0.000 description 27
- 230000006872 improvement Effects 0.000 description 19
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 15
- 229940080701 chymosin Drugs 0.000 description 14
- 230000000694 effects Effects 0.000 description 12
- 239000000463 material Substances 0.000 description 12
- 230000000052 comparative effect Effects 0.000 description 10
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- 231100000753 hepatic injury Toxicity 0.000 description 8
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- 102000004169 proteins and genes Human genes 0.000 description 8
- 108090000623 proteins and genes Proteins 0.000 description 8
- 238000002474 experimental method Methods 0.000 description 7
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- 102000004196 processed proteins & peptides Human genes 0.000 description 6
- 239000000047 product Substances 0.000 description 5
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 4
- 206010061218 Inflammation Diseases 0.000 description 4
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- 230000000050 nutritive effect Effects 0.000 description 4
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- 230000015572 biosynthetic process Effects 0.000 description 3
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- 108010009736 Protein Hydrolysates Proteins 0.000 description 2
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- 230000010030 glucose lowering effect Effects 0.000 description 2
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
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- 239000013642 negative control Substances 0.000 description 2
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- 238000012360 testing method Methods 0.000 description 2
- UUUHXMGGBIUAPW-UHFFFAOYSA-N 1-[1-[2-[[5-amino-2-[[1-[5-(diaminomethylideneamino)-2-[[1-[3-(1h-indol-3-yl)-2-[(5-oxopyrrolidine-2-carbonyl)amino]propanoyl]pyrrolidine-2-carbonyl]amino]pentanoyl]pyrrolidine-2-carbonyl]amino]-5-oxopentanoyl]amino]-3-methylpentanoyl]pyrrolidine-2-carbon Chemical compound C1CCC(C(=O)N2C(CCC2)C(O)=O)N1C(=O)C(C(C)CC)NC(=O)C(CCC(N)=O)NC(=O)C1CCCN1C(=O)C(CCCN=C(N)N)NC(=O)C1CCCN1C(=O)C(CC=1C2=CC=CC=C2NC=1)NC(=O)C1CCC(=O)N1 UUUHXMGGBIUAPW-UHFFFAOYSA-N 0.000 description 1
- 208000007848 Alcoholism Diseases 0.000 description 1
- 208000004930 Fatty Liver Diseases 0.000 description 1
- 229940123457 Free radical scavenger Drugs 0.000 description 1
- 108010024636 Glutathione Proteins 0.000 description 1
- 206010018910 Haemolysis Diseases 0.000 description 1
- 206010019708 Hepatic steatosis Diseases 0.000 description 1
- 108010061951 Methemoglobin Proteins 0.000 description 1
- 102000004270 Peptidyl-Dipeptidase A Human genes 0.000 description 1
- 108090000882 Peptidyl-Dipeptidase A Proteins 0.000 description 1
- 240000009235 Rubia cordifolia Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000009098 adjuvant therapy Methods 0.000 description 1
- 201000007930 alcohol dependence Diseases 0.000 description 1
- 230000001476 alcoholic effect Effects 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 235000021245 dietary protein Nutrition 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 208000010706 fatty liver disease Diseases 0.000 description 1
- 229960003180 glutathione Drugs 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000008588 hemolysis Effects 0.000 description 1
- 239000000413 hydrolysate Substances 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 238000011502 immune monitoring Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000021633 leukocyte mediated immunity Effects 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 230000003908 liver function Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
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- 230000036961 partial effect Effects 0.000 description 1
- 238000011056 performance test Methods 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 239000002516 radical scavenger Substances 0.000 description 1
- 150000003254 radicals Chemical class 0.000 description 1
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- 230000008439 repair process Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000012898 sample dilution Substances 0.000 description 1
- 230000002000 scavenging effect Effects 0.000 description 1
- BHZOKUMUHVTPBX-UHFFFAOYSA-M sodium acetic acid acetate Chemical compound [Na+].CC(O)=O.CC([O-])=O BHZOKUMUHVTPBX-UHFFFAOYSA-M 0.000 description 1
- 231100000240 steatosis hepatitis Toxicity 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
Classifications
-
- 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
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- 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
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/03—Organic compounds
-
- 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
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/30—Foods or foodstuffs containing additives; Preparation or treatment thereof containing carbohydrate syrups; containing sugars; containing sugar alcohols, e.g. xylitol; containing starch hydrolysates, e.g. dextrin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/01—Hydrolysed proteins; Derivatives thereof
- A61K38/011—Hydrolysed proteins; Derivatives thereof from plants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- 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
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/02—Antidotes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- General Health & Medical Sciences (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Polymers & Plastics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Food Science & Technology (AREA)
- Diabetes (AREA)
- Nutrition Science (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Immunology (AREA)
- Gastroenterology & Hepatology (AREA)
- Hematology (AREA)
- Botany (AREA)
- Endocrinology (AREA)
- Emergency Medicine (AREA)
- Toxicology (AREA)
- Obesity (AREA)
- Epidemiology (AREA)
- Mycology (AREA)
- Pain & Pain Management (AREA)
- Rheumatology (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention provides a plant peptide, which is prepared from the following raw materials in parts by weight: 70-80 parts of soybean peptide, 4-10 parts of corn oligopeptide, 10-20 parts of crystalline fructose and 1-2 parts of food additive. The plant peptide is characterized by being prepared from the following raw materials in parts by weight: 70-80 parts of soybean peptide, 4-10 parts of corn oligopeptide, 10-20 parts of crystalline fructose and 1-2 parts of food additive.
Description
Technical Field
The invention relates to the technical field of food, in particular to a plant peptide and a preparation method and application thereof.
Background
The corn oligopeptide powder is used as a mixture of various small peptides obtained by degrading corn protein through enzyme, has the excellent characteristics of peptide substances, namely the characteristics of direct absorption superior to amino acid or protein, strong solubility (the corn oligopeptide powder can be completely dissolved in water under a large pH value, no turbidity and precipitate are generated), strong stability (thermal stability, unchanged components, no loss of functions), high safety (natural food protein, safety, reliability, no toxic or side effect) and the like, and also has unique special functions.
The plant peptide refers to peptide substances derived from plants, and comprises peptides prepared by hydrolyzing, separating and purifying plants and peptides naturally existing in plants. In recent years, the research shows that the plant peptide has a plurality of biological functions of 1 and anti-tumor effect. Rubicin, for example, is a group of cyclohexapeptides with anticancer activity found in the plant rubia cordifolia. In addition, some polypeptide substances obtained from natural products have been reported in the literature to enhance cell-mediated immune response, improve immune self-stability and immune monitoring function. 2. Reducing blood pressure. The blood pressure lowering function of the plant peptide is achieved by inhibiting the activity of angiotensin converting enzyme. 3. Scavenging free radicals. As a free radical scavenger, it can protect cell membranes from oxidative damage, prevent hemolysis of erythrocytes, and promote the reduction of methemoglobin, such as glutathione.
Disclosure of Invention
The invention provides a plant peptide, a preparation method and application thereof, and aims to provide a plant peptide which is simple in preparation method, high in reaction efficiency, short in synthesis time, low in production cost, wide in raw material source and wide in application prospect, and the activity of a product is improved.
The invention provides a plant peptide which is prepared from the following raw materials in parts by weight: 70-80 parts of soybean peptide, 4-10 parts of corn oligopeptide, 10-20 parts of crystalline fructose and 1-2 parts of food additive.
As a further improvement of the invention, the food additive is prepared by mixing the following raw materials in parts by weight: 0.5 part of sweet orange essence, 0.5 part of bitter taste reducing essence, 0.4 part of mogroside and 0.05 part of stevioside.
As a further improvement of the invention, the soybean oligopeptide is obtained by carrying out enzymolysis on a soybean peptide raw material, the soybean oligopeptide is mixed with the corn oligopeptide, dissolved and adjusted in pH value, a protein-like reaction is carried out under certain conditions to obtain a composite peptide, and the composite peptide is added with crystalline fructose and a food additive and uniformly mixed to obtain the plant peptide.
As a further improvement of the invention, the preparation method of the soybean oligopeptide comprises the following steps: dissolving soybean peptide in water, adjusting pH of the solution to 8.5, adding alkaline protease for enzymolysis, adjusting pH to 7, adding neutral protease for enzymolysis, inactivating enzyme after enzymolysis, centrifuging, collecting supernatant, ultrafiltering, and lyophilizing to obtain soybean oligopeptide.
As a further improvement of the invention, the addition amount of the alkaline protease is 2500U/kg.
As a further improvement of the invention, the ultrafiltration is ultrafiltration by an ultrafiltration membrane with the molecular weight cut-off of 1000u, and the filtrate with the molecular weight of less than 1000u is obtained.
As a further improvement of the invention, the conditions of the protein-like reaction are that the pH is adjusted to 5-7, α -chymotrypsin and flavourzyme are added for reaction, after the reaction is finished, the enzyme is deactivated, the centrifugation is carried out, the supernatant is taken, dialyzed and freeze-dried, and the composite peptide is obtained.
As a further improvement of the invention, the addition amount of the α -chymochymotrypsin is 2000U/kg, and the addition amount of the flavourzyme is 3000U/kg.
The term "U/kg" as used herein means the activity (U) of the enzyme added to the material per kg.
As a further improvement of the invention, the freeze-drying condition is pre-cooling for 30-50min at-10 ℃, and then freeze-drying for 15-24h at-30 ℃.
The invention further protects the application of the plant peptide, and the plant peptide is safe and nontoxic and can be used for clinical related application, disease treatment and adjuvant treatment as a raw material of medicaments for relieving alcoholism and sobering up, resisting inflammation and protecting liver and reducing blood sugar, health-care food and functional food.
The invention has the following beneficial effects: the invention further cuts the soybean peptide into small molecular soybean oligopeptide by adopting a two-stage enzymolysis method, thereby improving the nutritive value of the soybean oligopeptide, and particularly having better anti-inflammatory, antioxidant and immunity-improving effects;
according to the invention, a protein-like reaction is utilized, soybean oligopeptide and corn oligopeptide are combined, the peptide chain growth is promoted, a mixture which has a higher molecular weight and is similar to protein is formed, the bitter taste in protein hydrolysate is eliminated, the nutritive value and the efficacy of the protein are improved, the specific biological activity of the protein in various aspects of reducing blood sugar, resisting inflammation, improving immunity and the like is realized, and the liver-protecting and alcohol-dispelling effects of the corn oligopeptide are enhanced;
the preparation method of the plant peptide is simple, the reaction efficiency is high, the synthesis time is short, the production cost is reduced, the activity of the product is improved, the raw material source is wide, and the application prospect is wide
Detailed Description
The technical solutions in the embodiments of the present invention are clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The freeze-drying conditions of the soybean oligopeptide in the embodiment and the comparative example are as follows: precooling for 30min at the temperature of minus 10 ℃, and then freeze-drying for 12h at the temperature of minus 30 ℃; inactivating enzyme at 105 deg.C for 15min, centrifuging at 10000r/min for 10 min. The dialysis conditions were 1 day by using a semipermeable membrane.
Example 1
The raw materials comprise the following components in parts by weight: 70 parts of soybean peptide, 4 parts of corn oligopeptide, 10 parts of crystalline fructose and 1 part of food additive.
The food additive is prepared by mixing the following raw materials in parts by weight: 0.5 part of sweet orange essence, 0.5 part of bitter taste reducing essence, 0.4 part of mogroside and 0.05 part of stevioside.
A preparation method of plant peptide comprises the following steps:
s1, dissolving soybean peptide in water (the mass ratio of the soybean peptide to the water is 1:10), adjusting the pH of the solution to 8.5 by using 1mol/L NaOH solution, adding alkaline protease for enzymolysis, then adjusting the pH to 7 by using 1mol/L HCl solution, adding neutral protease for enzymolysis, adding 2500U/kg of the neutral protease for enzymolysis, inactivating enzyme after the enzymolysis is finished, centrifuging, taking supernate, performing ultrafiltration by using an ultrafiltration membrane with the molecular weight cutoff of 1000U to obtain filtrate with the molecular weight of less than 1000U, and freeze-drying to obtain soybean oligopeptide;
s2, mixing soybean oligopeptide and corn oligopeptide, dissolving the mixture in water (the mass ratio of the materials to the water is 1:10), adjusting the pH to 5 by using 1mol/L HCl solution, adding α -chymosin and flavourzyme for reaction, wherein the addition amount of α -chymosin is 2000U/kg, the addition amount of flavourzyme is 3000U/kg, inactivating the enzyme after the reaction is finished, centrifuging, taking supernate, dialyzing, precooling for 30min at the temperature of-10 ℃, freeze-drying for 15h at the temperature of-30 ℃ to obtain composite peptide, adding crystalline fructose and a food additive, and uniformly mixing to obtain the plant peptide.
Example 2
The raw materials comprise the following components in parts by weight: 80 parts of soybean peptide, 10 parts of corn oligopeptide, 20 parts of crystalline fructose and 2 parts of food additive.
The food additive is prepared by mixing the following raw materials in parts by weight: 0.5 part of sweet orange essence, 0.5 part of bitter taste reducing essence, 0.4 part of mogroside and 0.05 part of stevioside.
A preparation method of plant peptide comprises the following steps:
s1, dissolving soybean peptide in water (the mass ratio of materials to water is 1:10), adjusting the pH of the solution to 8.5 by using 1mol/L NaOH solution, adding alkaline protease for enzymolysis, then adjusting the pH to 7 by using 1mol/L HCl solution, adding neutral protease for enzymolysis, adding 2500U/kg, inactivating enzyme after the enzymolysis is finished, centrifuging, taking supernate, performing ultrafiltration by using an ultrafiltration membrane with the molecular weight cutoff of 1000U to obtain filtrate with the molecular weight of less than 1000U, and freeze-drying to obtain soybean oligopeptide;
s2, mixing soybean oligopeptide and corn oligopeptide, dissolving the mixture in water (the mass ratio of the materials to the water is 1:10), adjusting the pH to 7 by using 1mol/L HCl solution, adding α -chymosin and flavourzyme for reaction, wherein the addition amount of α -chymosin is 2000U/kg, the addition amount of flavourzyme is 3000U/kg, inactivating the enzyme after the reaction is finished, centrifuging, taking supernate, dialyzing, precooling for 50min at the temperature of-10 ℃, freeze-drying for 24h at the temperature of-30 ℃ to obtain composite peptide, adding crystalline fructose and a food additive, and uniformly mixing to obtain the plant peptide.
Example 3
The raw materials comprise the following components in parts by weight: 75 parts of soybean peptide, 6 parts of corn oligopeptide, 15 parts of crystalline fructose and 1.5 parts of food additive.
The food additive is prepared by mixing the following raw materials in parts by weight: 0.5 part of sweet orange essence, 0.5 part of bitter taste reducing essence, 0.4 part of mogroside and 0.05 part of stevioside.
A preparation method of plant peptide comprises the following steps:
s1, dissolving soybean peptide in water (the mass ratio of materials to water is 1:10), adjusting the pH of the solution to 8.5 by using 1mol/L NaOH solution, adding alkaline protease for enzymolysis, then adjusting the pH to 7 by using 1mol/L HCl solution, adding neutral protease for enzymolysis, adding 2500U/kg, inactivating enzyme after the enzymolysis is finished, centrifuging, taking supernate, performing ultrafiltration by using an ultrafiltration membrane with the molecular weight cutoff of 1000U to obtain filtrate with the molecular weight of less than 1000U, and freeze-drying to obtain soybean oligopeptide;
s2, mixing soybean oligopeptide and corn oligopeptide, dissolving the mixture in water (the mass ratio of the materials to the water is 1:10), adjusting the pH to 6 by using 1mol/L HCl solution, adding α -chymosin and flavourzyme for reaction, wherein the addition amount of α -chymosin is 2000U/kg, the addition amount of flavourzyme is 3000U/kg, inactivating the enzyme after the reaction is finished, centrifuging, taking supernate, dialyzing, precooling for 40min at the temperature of-10 ℃, freeze-drying for 20h at the temperature of-30 ℃ to obtain composite peptide, adding crystalline fructose and a food additive, and uniformly mixing to obtain the plant peptide.
Comparative example 1
The raw materials comprise the following components in parts by weight: 75 parts of soybean peptide, 6 parts of corn oligopeptide, 15 parts of crystalline fructose and 1.5 parts of food additive.
The food additive is prepared by mixing the following raw materials in parts by weight: 0.5 part of sweet orange essence, 0.5 part of bitter taste reducing essence, 0.4 part of mogroside and 0.05 part of stevioside.
A preparation method of plant peptide comprises the following steps:
mixing soybean peptide and corn oligopeptide, adding crystalline fructose and food additive, and mixing to obtain the plant peptide.
Comparative example 2
The raw materials comprise the following components in parts by weight: 75 parts of soybean peptide, 6 parts of corn oligopeptide, 15 parts of crystalline fructose and 1.5 parts of food additive.
The food additive is prepared by mixing the following raw materials in parts by weight: 0.5 part of sweet orange essence, 0.5 part of bitter taste reducing essence, 0.4 part of mogroside and 0.05 part of stevioside.
A preparation method of plant peptide comprises the following steps:
mixing soybean peptide and corn oligopeptide, dissolving the mixture in water (the mass ratio of the materials to the water is 1:10), adjusting the pH to 6 by using 1mol/LHCl solution, adding α -chymosin and flavourzyme for reaction, wherein the addition amount of α -chymosin and flavourzyme is 2000U/kg, the addition amount of flavourzyme is 3000U/kg, inactivating enzyme after the reaction is finished, centrifuging, taking supernatant, dialyzing, precooling for 40min at-10 ℃, freeze-drying for 20h at-30 ℃ to obtain composite peptide, adding crystalline fructose and food additive, and uniformly mixing to obtain the plant peptide.
Comparative example 3
The raw materials comprise the following components in parts by weight: 6 parts of corn oligopeptide, 15 parts of crystalline fructose and 1.5 parts of food additive.
The food additive is prepared by mixing the following raw materials in parts by weight: 0.5 part of sweet orange essence, 0.5 part of bitter taste reducing essence, 0.4 part of mogroside and 0.05 part of stevioside.
A preparation method of plant peptide comprises the following steps:
dissolving corn oligopeptide in water (the mass ratio of the materials to the water is 1:10), adjusting the pH value to 6 by using 1mol/L HCl solution, adding α -chymosin and flavourzyme for reaction, wherein the addition amount of α -chymosin is 2000U/kg, the addition amount of flavourzyme is 3000U/kg, inactivating enzyme after the reaction is finished, centrifuging, taking supernate, dialyzing, precooling for 40min at the temperature of-10 ℃, freeze-drying for 20h at the temperature of-30 ℃, adding crystalline fructose and food additives, and uniformly mixing to obtain the plant peptide.
Comparative example 4
The raw materials comprise the following components in parts by weight: 75 parts of soybean peptide, 15 parts of crystalline fructose and 1.5 parts of food additive.
The food additive is prepared by mixing the following raw materials in parts by weight: 0.5 part of sweet orange essence, 0.5 part of bitter taste reducing essence, 0.4 part of mogroside and 0.05 part of stevioside.
A preparation method of plant peptide comprises the following steps:
s1, dissolving soybean peptide in water (the mass ratio of materials to water is 1:10), adjusting the pH of the solution to 8.5 by using 1mol/L NaOH solution, adding alkaline protease for enzymolysis, then adjusting the pH to 7 by using 1mol/L HCl solution, adding neutral protease for enzymolysis, adding 2500U/kg, inactivating enzyme after the enzymolysis is finished, centrifuging, taking supernate, performing ultrafiltration by using an ultrafiltration membrane with the molecular weight cutoff of 1000U to obtain filtrate with the molecular weight of less than 1000U, and freeze-drying to obtain soybean oligopeptide;
s2, dissolving soybean oligopeptide in water (the mass ratio of the materials to the water is 1:10), adjusting the pH value to 6 by using 1mol/L HCl solution, adding α -chymosin and flavourzyme for reaction, wherein the addition amount of α -chymosin and flavourzyme are 2000U/kg and 3000U/kg respectively, inactivating enzymes after the reaction is finished, centrifuging, taking supernate, dialyzing, precooling for 40min at the temperature of-10 ℃, freeze-drying for 20h at the temperature of-30 ℃, adding crystalline fructose and food additives, and uniformly mixing to obtain the plant peptide.
Comparative example 5
The raw materials comprise the following components in parts by weight: 20 parts of soybean peptide, 50 parts of corn oligopeptide, 15 parts of crystalline fructose and 1.5 parts of food additive.
The food additive is prepared by mixing the following raw materials in parts by weight: 0.5 part of sweet orange essence, 0.5 part of bitter taste reducing essence, 0.4 part of mogroside and 0.05 part of stevioside.
A preparation method of plant peptide comprises the following steps:
s1, dissolving soybean peptide in water (the mass ratio of materials to water is 1:10), adjusting the pH of the solution to 8.5 by using 1mol/L NaOH solution, adding alkaline protease for enzymolysis, then adjusting the pH to 7 by using 1mol/L HCl solution, adding neutral protease for enzymolysis, adding 2500U/kg, inactivating enzyme after the enzymolysis is finished, centrifuging, taking supernate, performing ultrafiltration by using an ultrafiltration membrane with the molecular weight cutoff of 1000U to obtain filtrate with the molecular weight of less than 1000U, and freeze-drying to obtain soybean oligopeptide;
s2, mixing soybean oligopeptide with corn oligopeptide, dissolving the mixture in water (the mass ratio of the materials to the water is 1:10), adjusting the pH value to 6, adding α -chymosin and flavourzyme for reaction, wherein the addition amount of α -chymosin is 2000U/kg, the addition amount of flavourzyme is 3000U/kg, inactivating enzyme after the reaction is finished, centrifuging, taking supernatant, dialyzing, precooling for 40min at the temperature of-10 ℃, freeze-drying for 20h at the temperature of-30 ℃ to obtain composite peptide, adding crystalline fructose and food additives, and uniformly mixing to obtain the plant peptide.
Test example 1 blood sugar lowering Performance test
The method for determining the activity of the DPP-4 inhibitory active peptide in vitro comprises the following steps:
the sample was diluted to an appropriate concentration with 100mmol/L Tris-HCl (pH8.0) buffer, 25. mu.L of the sample dilution was aspirated, mixed with 25. mu.L of a substrate (concentration: 1.6mmol/L), and added to a 96-well plate. After incubation for 10min at 37 ℃, adding 50 mu L of DPP-4 enzyme solution (the enzyme activity is 8U/L), evenly mixing, incubating for 60min at 37 ℃, immediately adding 100 mu L of 1mol/L acetic acid-sodium acetate (pH 4.0) buffer solution to terminate the reaction, measuring the absorbance A at 405nm, and calculating the DPP-4 inhibition rate of the hydrolysate according to the following formula. Each set of experiments was repeated 3 times.
DPP-4 inhibition (%) {1- (sample-a sample blank)/(a negative control-a negative blank) } × 100
Wherein: sample A: is the light absorption value A of the sample reaction solution at 405 nm;
sample a blank: taking Tris-HCL buffer solution as a sample blank control instead of DPP-4 enzyme solution, and taking the absorbance value A at 405 nm;
negative control A: taking Tris-HCL buffer solution as a negative control to replace the sample, and taking the absorbance A at 405 nm;
negative blank control a: Tris-HCl buffer was used as a negative blank, instead of DPP-4 enzyme solution and sample, absorbance A at 405 nm.
And (3) determining a sample: composite peptides prepared in examples 1 to 3 and comparative examples 1 to 5 and commercially available composite peptides (national treasure brand vitamin complex peptide powder, Nicoti New era health industry Co., Ltd.).
TABLE 1 DPP-4 inhibition ratio of composite peptide powder
As can be seen from Table 1, the composite peptide prepared in the examples 1 to 3 of the present invention has a blood glucose lowering function significantly superior to that of the same kind of commercially available products, and shows an increase in blood glucose lowering effect with an increase in concentration, but at a higher concentration, the increase is slower.
Compared with the embodiment 3, the soybean oligopeptide is not formed by the soybean peptide through enzymolysis, and meanwhile, the soybean peptide and the corn oligopeptide are not subjected to the growth of the protein-like reaction chain to form the protein-like factor with better blood sugar reducing effect, so the blood sugar reducing effect is greatly reduced.
Compared with the example 3, the soybean peptide of the comparative example 2 is not subjected to enzymolysis, but is subjected to partial enzymolysis during the later period of the protein-like reaction to form part of protein-like factors with better blood sugar reducing effect, but the blood sugar reducing effect is obviously inferior to that of the plant peptide in the example 3.
Comparative example 3 compared with example 3, the soybean peptide is not added in the raw materials, and the soybean oligopeptide is not involved in the protein-like reaction for forming the composite peptide, so that the protein-like factor with better blood sugar reducing effect is not formed under the formula
Compared with the example 3, the raw materials are not added with the corn oligopeptide, and the corn oligopeptide does not participate in the protein-like reaction for forming the composite peptide, so that the protein-like factor with better blood sugar reducing effect is not formed under the formula.
Compared with the example 3, the comparative example 5 has the same raw materials but different formula amounts, and the result shows that the blood sugar reducing effect is greatly reduced and is less than half of the original result, so that the protein-like factors with better blood sugar reducing effect cannot be formed under the formula ratio.
Test example 2 crowd efficacy experiment of liver-protecting formula food
The experimental population is as follows: the experiment is divided into 50-60 years old group, 60-70 years old group and 70-80 years old group, and each group selects 10 patients with liver injury such as fatty liver, alcoholic liver and the like with similar living habits and living standards.
The experimental method comprises the following steps: liver function tests were performed once on each experimental group prior to the start of the experiment and the results were recorded. The product is administered twice a day in a dose of 1 bag 10g each time, and is administered with warm water 30 days later, and liver function test is performed again, and compared with the result before the experiment, the improvement of liver injury is recorded.
The experimental results are as follows:
the improvement is better: the liver injury is obviously improved
The general improvement is as follows: slight improvement of liver injury
No improvement: without any improvement in the condition of liver injury
TABLE 2 results of the population observation experiment
| Better improve | General improvements | Without improvement | Total improvement (%) | |
| 50-60 years old group | 7 | 2 | 1 | 90 |
| 60-70 years old group | 6 | 3 | 1 | 90 |
| Group 70-80 years old | 6 | 2 | 2 | 80 |
As can be seen from the data in Table 1, the liver injury of each experimental group population is improved to different degrees, and the total improvement rate is 80% -90%, which shows that the invention can help to repair the liver injury, restore the liver function and has the function of preventing chemical liver injury.
Compared with the prior art, the invention further cuts the soybean peptide into small-molecular soybean oligopeptide by adopting a two-stage enzymolysis method, thereby improving the nutritive value of the soybean oligopeptide, and particularly having better effects of resisting inflammation and oxidation and improving the immunity;
according to the invention, a protein-like reaction is utilized, soybean oligopeptide and corn oligopeptide are combined, the peptide chain growth is promoted, a mixture which has a higher molecular weight and is similar to protein is formed, the bitter taste in protein hydrolysate is eliminated, the nutritive value and the efficacy of the protein are improved, the specific biological activity of the protein in various aspects of reducing blood sugar, resisting inflammation, improving immunity and the like is realized, and the liver-protecting and alcohol-dispelling effects of the corn oligopeptide are enhanced;
the preparation method of the plant peptide is simple, the reaction efficiency is high, the synthesis time is short, the production cost is reduced, the activity of the product is improved, the raw material source is wide, and the application prospect is wide.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (10)
1. The plant peptide is characterized by being prepared from the following raw materials in parts by weight: 70-80 parts of soybean peptide, 4-10 parts of corn oligopeptide, 10-20 parts of crystalline fructose and 1-2 parts of food additive.
2. The plant peptide according to claim 1, wherein the food additive is prepared by mixing the following raw materials in parts by weight: 0.5 part of sweet orange essence, 0.5 part of bitter taste reducing essence, 0.4 part of mogroside and 0.05 part of stevioside.
3. A process for preparing the plant peptide as claimed in claim 1 or 2, wherein the plant peptide is prepared from soybean peptide through enzymolyzing to obtain soybean oligopeptide, mixing with corn oligopeptide, dissolving in water, reaction to obtain composite peptide, adding crystalline fructose and food additive, and mixing.
4. The method for preparing the plant peptide according to claim 3, wherein the soybean oligopeptide is prepared by the following steps: dissolving soybean peptide in water, adjusting pH of the solution to 8-10, adding alkaline protease for enzymolysis, adjusting pH to 6.5-7.5, adding neutral protease for enzymolysis, inactivating enzyme after enzymolysis, centrifuging, collecting supernatant, ultrafiltering, and lyophilizing to obtain soybean oligopeptide.
5. The method for producing a plant peptide according to claim 4, wherein the amount of the alkaline protease added is 2500U/kg.
6. The method of claim 4, wherein the ultrafiltration is performed with an ultrafiltration membrane having a molecular weight cut-off of 1000u to obtain a filtrate having a molecular weight of less than 1000 u.
7. The method for preparing plant peptide according to claim 3, wherein the conditions of the proteinoid reaction are adjusted to pH 5-7, α -chymotrypsin and flavourzyme are added for reaction, after the reaction, enzyme deactivation and centrifugation are carried out, and the composite peptide is obtained by taking supernatant, dialyzing and freeze-drying.
8. The method for producing a plant peptide according to claim 7, wherein the α -chymotrypsin is added in an amount of 2000U/kg, and the flavourzyme is added in an amount of 3000U/kg.
9. The method for preparing the plant peptide according to claim 4 or 7, wherein the lyophilization condition is pre-cooling for 30-50min at-10 ℃, and then freeze-drying for 15-24h at-30 ℃.
10. Use of the plant peptide according to any one of claims 1 to 9 for preparing a medicament, health food or functional food for alleviating hangover, protecting liver, and lowering blood sugar.
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