CN114921514B - Hechong peptide and preparation method and application thereof - Google Patents
Hechong peptide and preparation method and application thereof Download PDFInfo
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- CN114921514B CN114921514B CN202210389573.2A CN202210389573A CN114921514B CN 114921514 B CN114921514 B CN 114921514B CN 202210389573 A CN202210389573 A CN 202210389573A CN 114921514 B CN114921514 B CN 114921514B
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- 238000002360 preparation method Methods 0.000 title claims abstract description 17
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- 241000209504 Poaceae Species 0.000 claims abstract description 53
- 239000004365 Protease Substances 0.000 claims abstract description 39
- 239000011550 stock solution Substances 0.000 claims abstract description 26
- 108091005804 Peptidases Proteins 0.000 claims abstract description 18
- 238000004140 cleaning Methods 0.000 claims abstract description 17
- 102000004142 Trypsin Human genes 0.000 claims abstract description 16
- 108090000631 Trypsin Proteins 0.000 claims abstract description 16
- 239000012588 trypsin Substances 0.000 claims abstract description 16
- 239000006228 supernatant Substances 0.000 claims abstract description 14
- 239000000047 product Substances 0.000 claims abstract description 13
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- OHDRQQURAXLVGJ-HLVWOLMTSA-N azane;(2e)-3-ethyl-2-[(e)-(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid Chemical compound [NH4+].[NH4+].S/1C2=CC(S([O-])(=O)=O)=CC=C2N(CC)C\1=N/N=C1/SC2=CC(S([O-])(=O)=O)=CC=C2N1CC OHDRQQURAXLVGJ-HLVWOLMTSA-N 0.000 description 9
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- OCZVHBZNPVABKX-UHFFFAOYSA-N 1,1-diphenyl-2-(2,4,6-trinitrophenyl)hydrazine;ethanol Chemical compound CCO.[O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1NN(C=1C=CC=CC=1)C1=CC=CC=C1 OCZVHBZNPVABKX-UHFFFAOYSA-N 0.000 description 2
- GLDQAMYCGOIJDV-UHFFFAOYSA-N 2,3-dihydroxybenzoic acid Chemical compound OC(=O)C1=CC=CC(O)=C1O GLDQAMYCGOIJDV-UHFFFAOYSA-N 0.000 description 2
- 102000000634 Cytochrome c oxidase subunit IV Human genes 0.000 description 2
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- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 2
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- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 230000031700 light absorption Effects 0.000 description 2
- FJKROLUGYXJWQN-UHFFFAOYSA-N papa-hydroxy-benzoic acid Natural products OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 description 2
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- WQGWDDDVZFFDIG-UHFFFAOYSA-N pyrogallol Chemical compound OC1=CC=CC(O)=C1O WQGWDDDVZFFDIG-UHFFFAOYSA-N 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 229960004889 salicylic acid Drugs 0.000 description 2
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- 235000013619 trace mineral Nutrition 0.000 description 2
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- 108010052832 Cytochromes Proteins 0.000 description 1
- 229910002567 K2S2O8 Inorganic materials 0.000 description 1
- 241000158764 Murraya Species 0.000 description 1
- 240000002853 Nelumbo nucifera Species 0.000 description 1
- 235000006508 Nelumbo nucifera Nutrition 0.000 description 1
- 241000243827 Nereis Species 0.000 description 1
- 241001248610 Ophiocordyceps sinensis Species 0.000 description 1
- 241000243803 Tylorrhynchus heterochaetus Species 0.000 description 1
- 102000006668 UniProt protein families Human genes 0.000 description 1
- 108020004729 UniProt protein families Proteins 0.000 description 1
- 241001661641 Verrucosa Species 0.000 description 1
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- 238000006701 autoxidation reaction Methods 0.000 description 1
- OHJMTUPIZMNBFR-UHFFFAOYSA-N biuret Chemical compound NC(=O)NC(N)=O OHJMTUPIZMNBFR-UHFFFAOYSA-N 0.000 description 1
- 125000002091 cationic group Chemical group 0.000 description 1
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Classifications
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- 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
- 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
-
- 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
- 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
- A61K38/012—Hydrolysed proteins; Derivatives thereof from animals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
-
- 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
- A61P39/06—Free radical scavengers or antioxidants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Animal Behavior & Ethology (AREA)
- Polymers & Plastics (AREA)
- Veterinary Medicine (AREA)
- Organic Chemistry (AREA)
- Public Health (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Food Science & Technology (AREA)
- Molecular Biology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Biochemistry (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Toxicology (AREA)
- Mycology (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Birds (AREA)
- Genetics & Genomics (AREA)
- Dermatology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Gerontology & Geriatric Medicine (AREA)
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- General Engineering & Computer Science (AREA)
- Animal Husbandry (AREA)
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- Cosmetics (AREA)
Abstract
The invention belongs to the technical field of polypeptide products, and discloses a graminide peptide, a preparation method and application thereof. The preparation method comprises the following steps: cleaning and crushing the gramineae, centrifuging, and collecting supernatant to obtain gramineae protein stock solution; adding protease into the stock solution of the graminaceous insect protein, carrying out enzymolysis, centrifuging, and collecting supernatant. According to the preparation method provided by the invention, trypsin, pepsin, papain and bromelain are adopted to carry out enzymolysis on broken graminaceous worms, and the graminaceous worms peptides are obtained through filtration and purification. The preparation process is simple, low in cost, high in yield and easy to realize industrialization. The graminium peptide has excellent oxidation resistance, has obvious scavenging effect on DPPH free radical, ABTS free radical, hydroxyl free radical and superoxide anion under low concentration, can be widely used in cosmetics, nutritional foods, functional foods, health-care products, feeds and preparing antioxidant medicines, and is beneficial to development and utilization of graminium.
Description
Technical Field
The invention belongs to the technical field of polypeptide products, and particularly relates to a graminide peptide and a preparation method and application thereof.
Background
The graminaceous, the academic name, the Nepala verrucosa, is produced in coastal salty and fresh water juncture areas, belongs to the phylum of Androzoon, the class of Murraya, the order of wandering, the Nereidae, and the Nereidae of the genus Nereis on systematic classification, is a widely distributed warm temperate zone and subtropical species, and is distributed in the places such as Nanjing, shanghai, fujian and Guangdong in China. The cereal worm has rich nutrition, is rich in protein, high fat and various trace elements, and the history of eating cereal worm can be traced to the clean generation. The cereal worm also has high medicinal value, has the effects of nourishing yin and tonifying yang, strengthening spleen, resisting oxidation, improving immunity, warming body, dispelling dampness and the like, is called as 'Cordyceps sinensis' in water, and is a good food and medicine. The Gramineae has wide application prospect in cosmetics, foods, medicines, health products and the like. At present, the gramineae is almost fully eaten, fresh gramineae or frozen gramineae is directly cooked, and is sometimes chopped into a small workshop of gramineae sauce, but the large-scale industry is not formed yet. In addition, the processing technology of the gramineae is less, and the research of other active ingredients is relatively poor. Because the fresh gramineae is not resistant to storage and inconvenient to transport, the current processing technology severely limits the circulation and further development and utilization of gramineae products.
The whole body of the Hechong is Bao, and researches show that the Hechong is rich in protein, high fat and various trace elements, and the Hechong extract has certain activity. However, how to refine the gramineae and prepare the gramineae product with high activity, so that the gramineae is more widely developed and utilized, and the method is still a problem to be solved.
Therefore, there is a need to provide a processed product of graminaceous worms which can be more widely developed and utilized.
Disclosure of Invention
The present invention aims to solve at least one of the technical problems in the prior art described above. Therefore, the invention provides a graminide peptide, a preparation method and application thereof. The graminia peptide provided by the invention has higher oxidation resistance, can be used as a raw material of medicines and cosmetics, and is beneficial to development and utilization of graminia.
The first aspect of the invention provides a preparation method of graminide.
Specifically, the preparation method of the graminide comprises the following steps:
(1) Cleaning and crushing the gramineae, centrifuging, and collecting supernatant to obtain gramineae protein stock solution;
(2) And adding protease into the graminium protein stock solution, carrying out enzymolysis, centrifuging, and collecting supernatant to obtain the graminium peptide.
Preferably, in step (1), the graminaceous insect is a fresh graminaceous insect or a frozen graminaceous insect.
Preferably, the graminaceous insect is an allergic graminaceous insect (i.e., an adult insect) in sexual maturity.
Preferably, in the step (1), the cleaning process is to clean with clear water to remove sediment impurities on the surface; then the gauze is fished out by a filter spoon and dried. The body of the gramineae is soft, and care should be taken during cleaning so as to avoid the cracking of the gramineae and the explosion of pulp.
Preferably, in step (1), the crushing is carried out by crushing the gramineae at a rotation speed of 500-2000 r/min.
Preferably, in the step (1), the centrifugation is performed at a rotation speed of 1000-10000r/min for 10-60min; further preferably, in step (1), the centrifugation is performed at a rotational speed of 3000 to 10000r/min for 10 to 30min.
Preferably, in step (2), the mass of the protease is 3% -60% of the mass of the graminium protein stock solution; further preferably, in step (2), the mass of the protease is 20% -40% of the mass of the graminium protein stock solution.
Preferably, in step (2), the protease is selected from at least one of trypsin, pepsin, papain or bromelain; further preferably, the protease is selected from at least one of trypsin, pepsin or papain; more preferably, the protease is trypsin and/or pepsin; most preferably, the protease is trypsin.
Preferably, the enzyme activity of the trypsin is 100-500U/mg; further preferably, the trypsin has an enzyme activity of 200-400U/mg.
Preferably, the pH value is controlled to 7.5-8.0 when the trypsin is adopted for enzymolysis.
Preferably, the enzyme activity of the pepsin is 100-500U/mg; further preferably, the pepsin has an enzyme activity of 200-400U/mg.
Preferably, the pH value is controlled to be 1.5-2.0 when the pepsin is adopted for enzymolysis.
Preferably, the enzyme activity of the papain is 500-1000U/mg; further preferably, the enzyme activity of the papain is 600-900U/mg.
Preferably, the pH value is controlled to be 6.5-7.0 when the papain is adopted for enzymolysis.
Preferably, the enzyme activity of the bromelain is 400-1600U/mg; further preferably, the bromelain has an enzyme activity of 500-1000U/mg.
Preferably, the pH value is controlled to 7.0-7.5 when the bromelain is adopted for enzymolysis.
Preferably, in the step (2), the enzymolysis temperature is 30-60 ℃, the enzymolysis temperature is determined according to the selected protease, and the optimal enzymolysis temperature of different proteases is selected.
Preferably, in the step (2), the enzymolysis time is 0.5-48h; further preferably, in the step (2), the enzymolysis time is 0.5-10h; more preferably, in step (2), the time of the enzymolysis is 1 to 5 hours.
Preferably, in the step (2), the centrifugation is performed at 5000-10000r/min for 10-60min; further preferably, in step (2), the centrifugation is performed at 5000-8000r/min for 10-30min.
In a second aspect, the invention provides a graminide.
Specifically, the graminium peptide is prepared by the preparation method, the scavenging rate of the graminium peptide to hydroxyl free radicals is more than 65%, the scavenging rate to superoxide anions is more than or equal to 50%, the scavenging rate to DPPH free radicals is more than 40%, and the scavenging rate to ABTS free radicals is more than 55% with the concentration of 0.5 mg/mL.
Preferably, the concentration of the graminide is 0.5mg/mL, the clearance rate of the graminide to hydroxyl radicals is more than 65%, the clearance rate to superoxide anions is more than or equal to 60%, the clearance rate to DPPH radicals is more than 80%, and the clearance rate to ABTS radicals is more than 55%.
Further preferably, the concentration of the graminide is 0.5mg/mL, the clearance rate of the graminide to hydroxyl radicals is more than 85%, the clearance rate to superoxide anions is more than or equal to 95%, the clearance rate to DPPH radicals is more than 80%, and the clearance rate to ABTS radicals is more than 65%.
The third aspect of the invention provides the use of graminide.
In particular to the application of the graminide in cosmetics. Such as essence, cream, facial mask, etc.
In particular to application of the graminide in foods and health products.
In particular to application of the graminide in preparing antioxidant medicaments.
Compared with the prior art, the invention has the following beneficial effects:
(1) According to the preparation method provided by the invention, trypsin, pepsin, papain and bromelain are adopted to carry out enzymolysis on broken graminaceous insects, and the graminaceous peptides are obtained through filtration and purification. The preparation process is simple, convenient to operate, low in cost, environment-friendly, high in yield and easy to realize industrialization.
(2) The graminium peptide prepared by the invention has excellent oxidation resistance, particularly the graminium peptide obtained by trypsin enzymolysis has obvious scavenging effect on DPPH free radical, ABTS free radical, hydroxyl free radical and superoxide anion under low concentration, can be widely used in cosmetics, nutritional foods, functional foods, health care products and feeds and preparing antioxidant medicines, and is beneficial to development and utilization of graminium.
Drawings
FIG. 1 is a mass spectrum Basepeak diagram of the Gramineae protein obtained in example 1.
Detailed Description
In order to make the technical solutions of the present invention more apparent to those skilled in the art, the following examples will be presented. It should be noted that the following examples do not limit the scope of the invention.
The starting materials, reagents or apparatus used in the following examples are all available from conventional commercial sources or may be obtained by methods known in the art unless otherwise specified.
Example 1
A method for preparing graminide, comprising the following steps:
(1) Cleaning treatment of gramineae
Repeatedly cleaning fresh Gramineae (adults) which are metamorphosis bodies in selective maturity stage) with clear water to remove surface sediment impurities and the like; then the gauze is fished up by a filter spoon and dried; because the body of the gramineae is soft, care should be taken during cleaning so as to avoid the cracking of the gramineae and the explosion of pulp.
(2) Extracting water-soluble graminia protein stock solution
Weighing 5g of Gramineae, adding 50mL of ultrapure water, crushing Gramineae with a stirrer at a rotating speed of 1200r/min, transferring the Gramineae mixed solution into a centrifuge tube, centrifuging for 20min at a rotating speed of 6000r/min, discarding the precipitate, and collecting the supernatant to obtain the water-soluble Gramineae protein stock solution. Protein concentration was measured using BCA protein assay kit, and the test result showed that the protein concentration of the graminiella protein stock was 3.57mg/mL. Mass spectrometry was performed on water-soluble graminaceous protein stocks, wherein the major steps of the large-scale protein identification (Shotgun) project procedure included: proteolysis, LC-MS/MS analysis, database retrieval, data analysis, experimental report and the like, and the mass spectrum Basepeak diagram is shown in figure 1. Meanwhile, a mass spectrum database is adopted to search a software MaxQuant 1.6.1.0 to compare protein databases (derived from Uniprot Protein Database and species Tylorrhynchus heterochaetus (Nelumbo nucifera) to obtain 25 protein sequences, and the fact that water-soluble graminium proteins mainly comprise extracellular globulin-2B, extracellular globulin-2A, extracellular globulin-2C, extracellular hemoglobin linker chain 1, cytochrome C oxidase subunit 2, extracellular hemoglobin linker chain 2, cytochrome B, NADH-ubiquitin oxidoreductase chain 5, NADH-ubiquitin oxidoreductase chain 3, NADH-ubiquitin oxidoreductase chain 1, cytochrome C oxidase subunit 1 (fragment) and the like is found, wherein the proportion of the cytochrome C oxidase is the largest, and the antioxidation of graminium proteins and graminium peptides possibly has a great relationship with cytochrome C oxidase.
(3) Enzymatic extraction of water-soluble graminaceous insect proteins
a, preparing an enzyme solution: 0.10g of trypsin (250U/mg) was weighed, 10mL of ultrapure water was added thereto, dissolved, and sterilized by filtration through a membrane having a pore size of 0.22 μm to obtain a protease solution of 10 mg/mL.
b, enzymolysis of water-soluble graminium protein: taking 5mL of the water-soluble graminium protein stock solution prepared in the step (2), adding 357 mu L of trypsin solution (the mass of protease solution accounts for 20% of the mass of the graminium protein stock solution), adjusting pH=7.5, adding NaOH solution dropwise during enzymolysis for 120min (the pH of a system is kept constant to be pH=7.5), carrying out boiling water bath for 10min after enzymolysis is finished, inactivating enzyme, centrifuging for 10min at 4000r/min, collecting supernatant, and obtaining graminium peptide, wherein the concentration of protein is measured to be 3.12mg/mL.
Protein concentration was measured by BCA method, and peptide bond of protein or polypeptide and Cu in alkaline environment 2+ Complexing and combining Cu 2 + Reduction to Cu 1+ (biuret reaction). BCA and Cu 1+ The combination forms a stable violet-blue complex, has a high light absorption value at 562nm and is proportional to the protein concentration.
Example 2
A method for preparing graminide, comprising the following steps:
(1) Cleaning treatment of gramineae
Repeatedly cleaning fresh Gramineae (adults) which are metamorphosis bodies in selective maturity stage) with clear water to remove surface sediment impurities and the like; then the gauze is fished up by a filter spoon and dried; because the body of the gramineae is soft, care should be taken during cleaning so as to avoid the cracking of the gramineae and the explosion of pulp.
(2) Extracting water-soluble graminia protein stock solution
Weighing 5g of Gramineae, adding 50mL of ultrapure water, crushing Gramineae with a stirrer at a rotating speed of 1200r/min, transferring the Gramineae mixed solution into a centrifuge tube, centrifuging for 20min at a rotating speed of 6000r/min, discarding the precipitate, and collecting the supernatant to obtain the water-soluble Gramineae protein stock solution. Protein concentration was measured using BCA protein assay kit, and the test result showed that the protein concentration of the graminiella protein stock was 3.57mg/mL.
(3) Enzymatic extraction of water-soluble graminaceous insect proteins
a, preparing an enzyme solution: 0.10g of pepsin (250U/mg) was weighed, 10mL of ultrapure water was added thereto, dissolved, and sterilized by filtration through a membrane having a pore size of 0.22 μm to obtain a protease solution of 10 mg/mL.
b, enzymolysis of water-soluble graminium protein: pepsin enzymolysis: taking 5mL of the water-soluble graminium protein stock solution prepared in the step (2), adding 535 mu L of pepsin solution (the mass of the protease solution accounts for 30% of the mass of the graminium protein stock solution), adjusting pH=1.5, carrying out water bath at 37 ℃ for 180min (the pH of a system is kept constant by dripping HCl in the enzymolysis process), carrying out boiling water bath for 10min after the enzymolysis is finished, inactivating enzyme, adjusting the pH value to 5.0, centrifuging for 10min at the rotating speed of 4000r/min, collecting supernatant, and obtaining graminium peptide, wherein the protein concentration of the graminium peptide is measured to be 2.73mg/mL.
Example 3
A method for preparing graminide, comprising the following steps:
(1) Cleaning treatment of gramineae
Repeatedly cleaning fresh Gramineae (adults) which are metamorphosis bodies in selective maturity stage) with clear water to remove surface sediment impurities and the like; then the gauze is fished up by a filter spoon and dried; because the body of the gramineae is soft, care should be taken during cleaning so as to avoid the cracking of the gramineae and the explosion of pulp.
(2) Extracting water-soluble graminia protein stock solution
Weighing 5g of Gramineae, adding 50mL of ultrapure water, crushing Gramineae with a stirrer at a rotating speed of 1200r/min, transferring the Gramineae mixed solution into a centrifuge tube, centrifuging for 20min at a rotating speed of 6000r/min, discarding the precipitate, and collecting the supernatant to obtain the water-soluble Gramineae protein stock solution. Protein concentration was measured using BCA protein assay kit, and the test result showed that the protein concentration of the graminiella protein stock was 3.57mg/mL.
(3) Enzymatic extraction of water-soluble graminaceous insect proteins
a, preparing an enzyme solution: 0.10g of papain (800U/mg) was weighed, 10mL of ultrapure water was added thereto, dissolved, and sterilized by filtration through a membrane having a pore size of 0.22 μm to obtain a protease solution of 10 mg/mL.
b, enzymolysis of water-soluble graminium protein: papain enzymolysis: taking 5mL of the water-soluble graminium protein stock solution prepared in the step (2), adding 357 mu L of papain solution (the mass of the papain solution accounts for 20% of the mass of the graminium protein stock solution), adjusting pH=6.5, carrying out water bath at 55 ℃ for 90min, carrying out boiling water bath for 10min after enzymolysis is finished, inactivating enzyme, centrifuging for 10min at the rotating speed of 4000r/min, collecting supernatant, and obtaining graminium peptide, wherein the protein concentration of the graminium peptide is measured to be 3.05mg/mL.
Example 4
A method for preparing graminide, comprising the following steps:
(1) Cleaning treatment of gramineae
Repeatedly cleaning fresh Gramineae (adults) which are metamorphosis bodies in selective maturity stage) with clear water to remove surface sediment impurities and the like; then the gauze is fished up by a filter spoon and dried; because the body of the gramineae is soft, care should be taken during cleaning so as to avoid the cracking of the gramineae and the explosion of pulp.
(2) Extracting water-soluble graminia protein stock solution
Weighing 5g of Gramineae, adding 50mL of ultrapure water, crushing Gramineae with a stirrer at a rotating speed of 1200r/min, transferring the Gramineae mixed solution into a centrifuge tube, centrifuging for 20min at a rotating speed of 6000r/min, discarding the precipitate, and collecting the supernatant to obtain the water-soluble Gramineae protein stock solution. Protein concentration was measured using BCA protein assay kit, and the test result showed that the protein concentration of the graminiella protein stock was 3.57mg/mL.
(3) Enzymatic extraction of water-soluble graminaceous insect proteins
a, preparing an enzyme solution: 0.10g of bromelain (600U/mg) was weighed, 10mL of ultrapure water was added thereto, dissolved, and sterilized by filtration through a membrane having a pore size of 0.22 μm to obtain a protease solution of 10 mg/mL.
b, enzymolysis of water-soluble graminium protein: bromelain enzymolysis: taking 5mL of the water-soluble graminium protein stock solution prepared in the step (2), adding 535 mu L of bromelain solution (the mass of the bromelain solution is 20% of the mass of the graminium protein stock solution), adjusting pH=7, carrying out water bath at 55 ℃ for 9h, carrying out boiling water bath for 10min after enzymolysis is finished, inactivating enzyme, centrifuging for 10min at the rotating speed of 4000r/min, collecting supernatant fluid, and obtaining graminium peptide, wherein the protein concentration of the graminium peptide is measured to be 2.83mg/mL.
Product effect test
The graminipeptide prepared in examples 1-4 was prepared into 0.5mg/mL graminipeptide solutions, a set of VE and VC standard solutions was used as positive controls, and the scavenging ability of the graminipeptide prepared in examples 1-4 against different free radicals (hydroxyl radical, superoxide anion, DPPH radical and ABTS radical) was measured.
Measurement of DPPH radical scavenging ability: 1, 1-diphenyl-2-trinitrophenylhydrazine (DPPH for short) is a stable long-life free radical, and the ethanol solution is dark purple and has strong absorption near 517 nm. When a free radical scavenger exists, the light absorption of the DPPH ethanol solution is weakened due to the single electron pairing with the free radical scavenger. The extent of discoloration of the DPPH ethanol solution is linearly related to the number of electrons it receives, and thus the ability of the test sample to scavenge free radicals, i.e., the magnitude of antioxidant activity, can be evaluated.
Determination of the ability of hydroxyl radical to scavenge: generating hydroxyl radicals by Fenton reaction: h 2 O 2 +Fe 2+ ＝-OH+H 2 O+Fe 3+ Salicylic acid is added into a reaction system, and hydroxyl radicals generated by Fenton reaction react with the salicylic acid to generate 2,3 dihydroxybenzoic acid with special absorption at 510 nm. If a test substance having a function of scavenging hydroxyl radicals is added to the reaction system, the generated hydroxyl radicals are reduced, and the amount of colored compounds generated is reduced accordingly. The absorbance of the reaction liquid containing the measured object is measured at 510nm by adopting a fixed reaction time method, and compared with a blank liquid to determine the scavenging effect of the measured object on the hydroxyl free radicals.
ABTS radical scavenging ability assay: the ABTS method is to oxidize the color-developing agent ABTS into green ABTS under the action of proper oxidant + The absorbance of ABTS measured at wavelength 405nm gives the total antioxidant capacity of the sample. Direct generation of stable cationic free radical ABTS with K2S2O8 and ABTS + Antioxidant substances and ABTS + The reaction occurs to fade the reaction system.
Determination of superoxide anion scavenger capability: the pyrogallol is subjected to autoxidation under the weak base condition, so that superoxide anions can be generated, and the self-oxidation product has a characteristic absorption peak at the wavelength of 319nm and can be detected by a spectrophotometer.
The results of the ability of the graminide peptides prepared in examples 1-4 to scavenge different free radicals are shown in Table 1.
TABLE 1
As shown in Table 1, the stock solution of the graminium protein provided by the invention has a certain oxidation resistance, but when the graminium peptide obtained by adopting the method provided by the invention is exquisite, the oxidation resistance is improved to different degrees. Especially, the graminide prepared by trypsin enzymolysis in example 1 has excellent scavenging ability on hydroxyl free radical, superoxide anion, DPPH free radical and ABTS free radical. The antioxidant is strong in oxidation resistance, and can be widely applied to cosmetics and health care products.
Claims (4)
1. The preparation method of the graminide is characterized by comprising the following steps:
(1) Cleaning and crushing the gramineae, centrifuging, and collecting supernatant to obtain gramineae protein stock solution;
(2) Adding protease into the graminium protein stock solution, carrying out enzymolysis, centrifuging, and collecting supernatant to obtain graminium peptide;
in the step (2), the protease is trypsin;
the mass of the trypsin accounts for 20% of the mass of the graminaceous insect protein stock solution;
the enzyme activity of the trypsin is 250U/mg;
the enzymolysis temperature is 37 ℃; the enzymolysis time is 2 hours; the pH value of the enzymolysis is 7.5.
2. A graminium peptide prepared by the preparation method of claim 1, wherein the graminium peptide has a hydroxyl radical clearance of more than 85%, a superoxide anion clearance of more than or equal to 95%, a DPPH radical clearance of more than 80%, and an ABTS radical clearance of more than 65% at a concentration of 0.5 mg/mL.
3. Use of a graminide according to claim 2 for the preparation of an antioxidant cosmetic and/or an antioxidant health product.
4. Use of a graminide according to claim 2 for the preparation of an antioxidant medicament.
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7060790B1 (en) * | 1997-12-17 | 2006-06-13 | Vlaams Interuniversitair Instituut Voor Biotechnologie Vzw | Peptides and nucleic acids derived from Eisenia foetida and the use thereof |
| CN106722934A (en) * | 2016-11-25 | 2017-05-31 | 济南睿呋康生物科技有限公司 | A kind of anti-ageing health food for containing peptide extract containing earthworm, clam worm |
| CN107821258A (en) * | 2017-11-07 | 2018-03-23 | 华南师范大学 | A kind of method of artificial propagation standing grain worm |
| CN107950617A (en) * | 2017-12-29 | 2018-04-24 | 广州合诚实业有限公司 | A kind of standing grain worm egg flavor bread and preparation method thereof |
| CN110093335A (en) * | 2019-05-05 | 2019-08-06 | 北京颢美细胞基因生物技术有限公司 | A kind of extracting method of Nereid kinase |
| CN110904180A (en) * | 2019-12-23 | 2020-03-24 | 山东省海洋生物研究院 | Nereid antioxidant peptide and preparation method and application thereof |
| CN111647094A (en) * | 2020-06-11 | 2020-09-11 | 福建省水产研究所（福建水产病害防治中心） | Method for extracting nereis polysaccharide with auxiliary protection effect on chemical liver injury |
-
2022
- 2022-04-14 CN CN202210389573.2A patent/CN114921514B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7060790B1 (en) * | 1997-12-17 | 2006-06-13 | Vlaams Interuniversitair Instituut Voor Biotechnologie Vzw | Peptides and nucleic acids derived from Eisenia foetida and the use thereof |
| CN106722934A (en) * | 2016-11-25 | 2017-05-31 | 济南睿呋康生物科技有限公司 | A kind of anti-ageing health food for containing peptide extract containing earthworm, clam worm |
| CN107821258A (en) * | 2017-11-07 | 2018-03-23 | 华南师范大学 | A kind of method of artificial propagation standing grain worm |
| CN107950617A (en) * | 2017-12-29 | 2018-04-24 | 广州合诚实业有限公司 | A kind of standing grain worm egg flavor bread and preparation method thereof |
| CN110093335A (en) * | 2019-05-05 | 2019-08-06 | 北京颢美细胞基因生物技术有限公司 | A kind of extracting method of Nereid kinase |
| CN110904180A (en) * | 2019-12-23 | 2020-03-24 | 山东省海洋生物研究院 | Nereid antioxidant peptide and preparation method and application thereof |
| CN111647094A (en) * | 2020-06-11 | 2020-09-11 | 福建省水产研究所（福建水产病害防治中心） | Method for extracting nereis polysaccharide with auxiliary protection effect on chemical liver injury |
Non-Patent Citations (4)
| Title |
|---|
| Antifatigue effects of Hechong (Tylorrhynchus heterochaetus) through modulation of Nrf2/ARE- mediated antioxidant signaling pathway;Sunil等;Food and Chemical Toxicology;第157卷(第01期);1-8 * |
| 沙蚕多肽提取及功能研究;李雪等;食品安全质量检测学报;第09卷(第08期);1971-1975 * |
| 管华诗主编.《中华海洋本草精选本》.上海科学技术出版社,2014,(第01版),160-161. * |
| 罗永康主编.《生物活性肽功能与制备》.中国轻工业出版社,2019,(第01版),66. * |
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