CN108077939A - The deep-sea fish flesh of fish prepares the purposes of active peptides compound - Google Patents
The deep-sea fish flesh of fish prepares the purposes of active peptides compound Download PDFInfo
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- CN108077939A CN108077939A CN201711228159.9A CN201711228159A CN108077939A CN 108077939 A CN108077939 A CN 108077939A CN 201711228159 A CN201711228159 A CN 201711228159A CN 108077939 A CN108077939 A CN 108077939A
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- 241000251468 Actinopterygii Species 0.000 title claims abstract description 91
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 71
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 69
- 150000001875 compounds Chemical class 0.000 title claims abstract description 36
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- 150000003722 vitamin derivatives Chemical class 0.000 claims abstract description 6
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- 239000007788 liquid Substances 0.000 claims description 24
- 238000004108 freeze drying Methods 0.000 claims description 22
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- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 1
- 241001474374 Blennius Species 0.000 description 1
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- -1 methyl mercaptan compound Chemical class 0.000 description 1
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- 239000013589 supplement Substances 0.000 description 1
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- 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
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/04—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from fish or other sea animals
-
- 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
- A23L33/115—Fatty acids or derivatives thereof; Fats or oils
-
- 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
- A23L33/15—Vitamins
-
- 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
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Food Science & Technology (AREA)
- Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Mycology (AREA)
- Zoology (AREA)
- Biochemistry (AREA)
- Marine Sciences & Fisheries (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The active peptides compound prepared with the deep-sea fish flesh of fish for health products or drug, is played the role of extending the brain cell service life by the present invention.Above-mentioned active peptides are using the deep-sea fish flesh of fish as raw material, carry out stepwise discretization to it using Porphyra yezoensis protease and flavor protease and obtain.The present invention by 10 ~ 20 parts of active peptides, 5 ~ 10 parts of collagen small peptides, 20 ~ 30 parts of vitamin Cs and 3 ~ 5 parts of DHA by being mixed and made into pulvis or tablet or liquor obtains active peptides compound.It has the beneficial effect that：The active peptides low fat being prepared, no fishy smell, purity are high.Each ingredient has coupling in compound, can effectively inhibit the generation of endogenous endonuclease, delays the DNA degradation of cell chromosome, so as to delay the expression of apoptosis program in neuronal cell, effectively extend its service life.
Description
Technical field
It is specifically a kind of to prepare active peptides compound with the deep-sea fish flesh of fish the present invention relates to health products and medicine field
Purposes.
Background technology
Polypeptide attracts attention since earlier 1900s are just, and research finds that it is different from protein and is different from amino acid, tool
There is drug.Having arrived the last century nineties just has tremendous development, under the technological guidance of continuous renewal, polypeptide
Application range is more and more wider, and effect is also increasing, this has just strided forward our life, and the share occupied in we live is got over
Come bigger, polypeptide drug high-efficiency low-toxicity high specificity has the peculiar advantage of other drugs of being different from, and just constantly affects me
Life.And polypeptide has a high potential, and is the another quantum jump and developing direction of bioscience development.
Naturally occurring Marine Bioactive Peptides due in organism content it is low, and extract difficult, it is difficult to realize a large amount of
Needed for production supply, chemical artificial synthetic polypeptide is expensive.So sight is more invested exploitation proteolysis production by people
Come in this approach of object acquisition active peptides.Since the environment and terrestrial life of marine organisms existence are entirely different, such as high pressure, low
Temperature, high temperature and the extreme environments such as with high salt, in order to adapt to these extreme marine biological environments, marine protein matter no matter amino
The composition or the sequence of amino acid of acid are all very different with terrestrial life albumen, meanwhile, marine protein resource without
By being all far longer than land protein resource in type and quantity, and do not developed well.Miscellaneous ocean egg
In casamino acid sequence, potential many biologically active amino acid sequences with special protease hydrolytic, just release
Active peptide fragment.Marine protein resource is the important of the important albumen group food of the 21 century mankind and bioactive substance
Source.The current marine protein total resources in China comes out at the top in countries in the world, but the protide aquatic products that China is current
In product, in addition to a part is directly edible, major part is made food by simple processing technology and enters market, and processing technology falls
Afterwards, product structure is single, and value-added content of product is low so that the competitiveness of product in the international market is poor.Therefore, China is badly in need of
Marine protein resource is optimized and is utilized and high-valued processing.
Brain cell is the common name for the various kinds of cell for forming brain.Brain cell mainly includes neuron and Deiter's cells.God
Break up through first cell height, non-renewable, scientific research is shown, people can gradually decrease to brain cell number after 40 years old, generally
People can consume 10% or so to brain cell during death.Brain is administered activities, the cerebellums such as language, thinking memory and calculating and is known
The action of human body and the coordination ability, the reduction of brain cell can cause people the symptoms such as decrease of memory, hypophrenia occur.
The prior art such as Authorization Notice No. is the Chinese invention patent of CN101503728B, discloses one kind and accounts for fish fish from green grass or young crops
The method that composite polypeptide liquid is extracted in bone.Comprise the following steps：Mackerel bone is cleaned, is rubbed, is impregnated with deionized water；It crosses
Deionized water is filtered out, by fish-bone by the addition water of 1: 5-1: 10 mass ratio, animal is pressed by the 1%-5% amounts of fish-bone quality
The ratio that the mass ratio of proteolytic enzyme and neutral proteinase is 3: 1-1: 3 adds in enzyme, pH is adjusted in 5.5-8.5, then in 45-
It is digested at a temperature of 70 DEG C；Above-mentioned 90-100 DEG C of enzymolysis liquid is heated, so that enzyme inactivates；It is filtered with linen, filtrate is in 2000-
4000rpm centrifuges 10-15min, and obtained supernatant refilters degerming and obtain composite polypeptide liquid by GPC gel purifications.It is above-mentioned
The complex polypeptide purity arrived prepared by preparation method is not high, and loss amount is big in preparation process, and recovery rate is not high.Protease is equal
Using land protease, it is impossible to which marine protein is developed well.
The content of the invention
It is an object of the invention to provide one kind for health products or drug, play the role of extending using deeply for brain cell service life
The ocean fish flesh of fish prepares active peptides compound.The active peptides purity that the present invention is prepared is high, and each ingredient has coupling in product
Effect can effectively extend the service life of human body brain cell.
The problem of present invention in background technology for mentioning, the technical solution taken is：Activity is prepared with the deep-sea fish flesh of fish
The purposes of polypeptide compound for health products or drug, plays the role of extending the brain cell service life.
The preparation method of active peptides compound is prepared with the deep-sea fish flesh of fish, is comprised the following steps：
1）Degreasing：The deep-sea fish flesh of fish is cut into small pieces, and adds in 2 ~ 4 ‰ chitosan-immobilized lipase, adds in the steaming of 8 ~ 15 times of volumes
Distilled water, constant temperature digests 0.8 ~ 2.5h at 25 ~ 50 DEG C, and the degreasing deep-sea fish flesh of fish is obtained by filtration in enzyme deactivation after enzymolysis.Lipase
Fat in the deep-sea fish flesh of fish can be progressively hydrolyzed into glycerine and aliphatic acid.Lipase after immobilization, stability, enzyme activity
Property, heat resistance are significantly improved, and reusable；
2）Deodorant：The degreasing deep-sea fish flesh of fish is dipped in 0.4 ~ 0.8h in deodorant liquid, soaking temperature is 0 ~ 4 DEG C；Deodorant liquid ingredient and
Its parts by weight is 0.2 ~ 0.8 part of 3 ~ 8 parts of tea polyphenols, 10 ~ 20 parts of pure water and grape seed extract.Ingredient cooperates in above-mentioned deodorant liquid
Effect has various biological activity, can eliminate methyl mercaptan compound, and can be combined with amino acid, has centainly
Enzyme and bactericidal effect are passivated, and ingredient is natural, it is green safe；
3）Stepwise discretization：Minced fillet is made in deep-sea fish flesh of fish homogenate after deodorant, adds in 20 ~ 30 times of volume of water, 5 ~ 8wt, ‰ streaks
Seaweed protease, ethyl caprate and DTT.Wherein, the addition of ethyl caprate and DTT are respectively Porphyra yezoensis protease quality
1 ~ 5% and 10 ~ 20%.25 ~ 35 DEG C of hydrolysis temperature, pH are 7.0 ~ 9.0, and enzymolysis time is 30 ~ 40min, enzyme deactivation after enzymolysis；Add
Enter 1 ~ 3wt ﹪ flavor proteases, enzymolysis pH is 6.0 ~ 7.0, and hydrolysis temperature is 52 ~ 60 DEG C, and enzymolysis time is 15 ~ 25min.Deep-sea
There is the restriction enzyme site of a large amount of Porphyra yezoensis protease on Fish protein, can obtain peptide chain different in size after enzymolysis, then with flavor egg
White enzyme enzymolysis can obtain the active peptides that can effectively extend the brain cell service life.It cannot be obtained using other protease stepwise discretizations
State active peptides.Ethyl caprate and DTT are attached to the surface of Porphyra yezoensis protease by Van der Waals force and electrostatic interaction, one
Determine to change its tertiary structure in degree, the yield of active peptides can be also improved while the activity of Porphyra yezoensis is improved, but have
Body mechanism is still not clear；
4）Isoelectric precipitation：Enzymolysis liquid is filtered, the pH value for adjusting filtrate is 4.0 ~ 4.5, and centrifuging and taking precipitates after standing.Above-mentioned
Under pH environment, active peptides are in electroneutral, are slightly soluble in water, most of to be separated with precipitation form from aqueous solution；
5）Second ultrafiltration and nanofiltration：5 ~ 10 times of distilled water are added in the precipitation obtained in isoelectric point precipitation, after being completely dissolved first
Using the film ultrafiltration of retention 30 ~ 40kD of relative molecular mass, then the film ultrafiltration using 4 ~ 8kD of retention relative molecular mass, finally
Filtered solution is subjected to nanofiltration desalination.Be not in blocked up gel layer under above-mentioned condition, in ultra-filtration process, ultrasiltrated rate is also more
It is preferable；
6）Macroporous resin adsorption elutes：By the liquid concentration after nanofiltration, sample concentration is 0.1 ~ 0.3g/mL, and loading flow velocity is 0.4
~ 0.6mL/min, eluant, eluent volume are 7.7 ~ 7.9BV, and eluant, eluent is distilled water, and elution flow rate is 0.8 ~ 1.4mL/min.Macropore
Resin carries out adsorption and desorption to active peptides, and equilibrium adsorption capacity reaches up to 14.23mg/g or so, Static Adsorption rate
90.15% or so reaches 86.54% or so with static desorption efficiency, can effectively separate active peptides and other impurities；
7）Freeze-drying：The eluent of 10 ~ 30min periods is collected, is freeze-dried after its low temperature is concentrated, pre-freeze：-35~-45
DEG C 3 ~ 5h of pre-freeze；Freeze-drying：- 45 ~ -35 DEG C, 3 ~ 5Pa is freeze-dried 5 ~ 7h, and -10 ~ -5 DEG C, 10 ~ 20Pa is freeze-dried 2 ~ 3h；
Lyophilization：10 ~ 15 DEG C, 50 ~ 60Pa lyophilizations, 3 ~ 4h；Parsing-desiccation:7 ~ 14 DEG C, 3 ~ 5Pa, 2 ~ 4h of parsing-desiccation are lived
Property polypeptide.Lyophilized whole process is all under program control, easy to operate, and process is succinct, and high degree of automation reduces people
Power, material resources.Under low temperature drying can preferably retentive activity polypeptide activity；Dry under low pressure, active peptides are not oxidizable, curative effect
More preferably；Active peptides form " skeleton " when freezing, and original shape is preserved after dry, are basically unchanged for porous loose structure and color, produce
Product appearance is more excellent；Rehydration is good, and freeze drying activity polypeptide, which can absorb water rapidly, is reduced into lyophilized preceding state, easy to use；Dehydration
Thoroughly, water content is low in active peptides, generally in 1%-3%, and is conducive to long-distance transport and long-term preservation；
8）The preparation of compound：According to 10 ~ 20 parts of active peptides, 5 ~ 10 parts of collagen small peptides, 20 ~ 30 parts of vitamin Cs and 3 ~ 5
Part DHA is mixed and made into pulvis or tablet or liquor obtains active peptides compound.Each ingredient has coupling in above-mentioned compound,
The generation of endogenous endonuclease can effectively be inhibited, delay the DNA degradation of cell chromosome, so as to delay in neuronal cell
The expression of apoptosis program effectively extends its service life.
Compared with prior art, the advantage of the invention is that：The active peptides that the present invention is prepared with the deep-sea fish flesh of fish compound
Object is used for health products or drug, plays the role of extending the brain cell service life.Using Porphyra yezoensis protease and flavor protease to depth
The ocean fish flesh of fish carries out stepwise discretization, the active peptides low fat being prepared, no fishy smell, purity height.Each ingredient has coupling in compound
Cooperation is used, and can effectively be inhibited the generation of endogenous endonuclease, be delayed the DNA degradation of cell chromosome, so as to delay nerve
The expression of apoptosis program in first cell, effectively extends its service life.
Specific embodiment
The present invention program is described further below by embodiment：
Embodiment 1：
The purposes of active peptides compound is prepared with the deep-sea fish flesh of fish, for health products or drug, plays and extends the brain cell service life
Effect.
The preparation method of active peptides compound is prepared with the deep-sea fish flesh of fish, is comprised the following steps：
1）Degreasing：The deep-sea fish flesh of fish is cut into small pieces, and adds in 3 ‰ chitosan-immobilized lipase, adds in the distilled water of 10 times of volumes,
Constant temperature digests 2.0h at 30 DEG C, and the degreasing deep-sea fish flesh of fish is obtained by filtration in enzyme deactivation after enzymolysis；
2）Deodorant：The degreasing deep-sea fish flesh of fish is dipped in 0.6h in deodorant liquid, soaking temperature is 2 DEG C；Deodorant liquid ingredient and its weight
Part is 0.5 part of 5 parts of tea polyphenols, 16 parts of pure water and grape seed extract；
3）Stepwise discretization：Minced fillet is made in deep-sea fish flesh of fish homogenate after deodorant, adds in 25 times of volume of water, ‰ Porphyra yezoensis of 6wt
Protease, ethyl caprate and DTT.Wherein, the addition of ethyl caprate and DTT are respectively 3% He of Porphyra yezoensis protease quality
15%.30 DEG C, pH 8.0, enzymolysis time 35min of hydrolysis temperature, enzyme deactivation after enzymolysis；2wt ﹪ flavor proteases are added in,
It is 6.5 to digest pH, and hydrolysis temperature is 55 DEG C, enzymolysis time 20min；
4）Isoelectric precipitation：Enzymolysis liquid is filtered, the pH value for adjusting filtrate is 4.2, and centrifuging and taking precipitates after standing；
5）Second ultrafiltration and nanofiltration：8 times of distilled water are added in the precipitation obtained in isoelectric point precipitation, are first used after being completely dissolved
Retain the film ultrafiltration of relative molecular mass 35kD, then the film ultrafiltration using retention relative molecular mass 6kD, finally by filtered solution into
Row nanofiltration desalination；
6）Macroporous resin adsorption elutes：By the liquid concentration after nanofiltration, sample concentration 0.2g/mL, loading flow velocity is 0.5mL/
Min, eluant, eluent volume are 7.8BV, and eluant, eluent is distilled water, elution flow rate 1.0mL/min；
7）Freeze-drying：The eluent of 10 ~ 30min periods is collected, is freeze-dried after its low temperature is concentrated, pre-freeze：- 40 DEG C pre-
Freeze 4h；Freeze-drying：- 35 DEG C, 2Pa freeze-dryings 6h, -7 DEG C, 15Pa is freeze-dried 2.5h；Lyophilization：12 DEG C, 55Pa liters
The dry 3.5h of China；Parsing-desiccation:12 DEG C, 4Pa parsing-desiccation 3h obtain active peptides；
8）The preparation of compound：According to 15 parts of active peptides, 7 parts of collagen small peptides, 27 parts of vitamin Cs and 4 parts of DHA mixing systems
Active peptides compound is obtained into pulvis.
Embodiment 2：
The purposes of active peptides compound is prepared with the deep-sea fish flesh of fish, for health products or drug, plays and extends the brain cell service life
Effect.
The preparation method of active peptides compound is prepared with the deep-sea fish flesh of fish, is comprised the following steps：
1）Degreasing：The deep-sea fish flesh of fish is cut into small pieces, and adds in 2 ‰ chitosan-immobilized lipase, adds in the distilled water of 12 times of volumes,
Constant temperature digests 2.5h at 35 DEG C, and the degreasing deep-sea fish flesh of fish is obtained by filtration in enzyme deactivation after enzymolysis；
2）Deodorant：The degreasing deep-sea fish flesh of fish is dipped in 0.7h in deodorant liquid, soaking temperature is 4 DEG C；Deodorant liquid ingredient and its weight
Part is 0.6 part of 6 parts of tea polyphenols, 20 parts of pure water and grape seed extract；
3）Stepwise discretization：Minced fillet is made in deep-sea fish flesh of fish homogenate after deodorant, adds in 20 times of volume of water, ‰ Porphyra yezoensis of 5wt
Protease, ethyl caprate and DTT.Wherein, the addition of ethyl caprate and DTT are respectively 4% He of Porphyra yezoensis protease quality
18%.30 DEG C, pH 8.0, enzymolysis time 35min of hydrolysis temperature, enzyme deactivation after enzymolysis；2wt ﹪ flavor proteases are added in,
It is 6.5 to digest pH, and hydrolysis temperature is 55 DEG C, enzymolysis time 20min；
4）Isoelectric precipitation：Enzymolysis liquid is filtered, the pH value for adjusting filtrate is 4.2, and centrifuging and taking precipitates after standing；
5）Second ultrafiltration and nanofiltration：8 times of distilled water are added in the precipitation obtained in isoelectric point precipitation, are first used after being completely dissolved
Retain the film ultrafiltration of relative molecular mass 35kD, then the film ultrafiltration using retention relative molecular mass 6kD, finally by filtered solution into
Row nanofiltration desalination；
6）Macroporous resin adsorption elutes：By the liquid concentration after nanofiltration, sample concentration 0.2g/mL, loading flow velocity is 0.5mL/
Min, eluant, eluent volume are 7.8BV, and eluant, eluent is distilled water, elution flow rate 1.0mL/min；
7）Freeze-drying：The eluent of 10 ~ 30min periods is collected, is freeze-dried after its low temperature is concentrated, pre-freeze：- 40 DEG C pre-
Freeze 4h；Freeze-drying：- 35 DEG C, 2Pa freeze-dryings 6h, -7 DEG C, 15Pa is freeze-dried 2.5h；Lyophilization：12 DEG C, 55Pa liters
The dry 3.5h of China；Parsing-desiccation:12 DEG C, 4Pa parsing-desiccation 3h obtain active peptides；
8）The preparation of compound：According to 15 parts of active peptides, 7 parts of collagen small peptides, 27 parts of vitamin Cs and 4 parts of DHA mixing systems
Piece agent obtains active peptides compound.
Embodiment 3：
The purposes of active peptides compound is prepared with the deep-sea fish flesh of fish, for health products or drug, plays and extends the brain cell service life
Effect.
The preparation method of active peptides compound is prepared with the deep-sea fish flesh of fish, is comprised the following steps：
1）Degreasing：The deep-sea fish flesh of fish is cut into small pieces, and adds in 4 ‰ chitosan-immobilized lipase, adds in the distilled water of 15 times of volumes,
Constant temperature digests 2.5h at 25 DEG C, and the degreasing deep-sea fish flesh of fish is obtained by filtration in enzyme deactivation after enzymolysis；
2）Deodorant：The degreasing deep-sea fish flesh of fish is dipped in 0.6h in deodorant liquid, soaking temperature is 4 DEG C；Deodorant liquid ingredient and its weight
Part is 0.5 part of 8 parts of tea polyphenols, 18 parts of pure water and grape seed extract；
3）Stepwise discretization：Minced fillet is made in deep-sea fish flesh of fish homogenate after deodorant, adds in 25 times of volume of water, ‰ Porphyra yezoensis of 5wt
Protease, ethyl caprate and DTT.Wherein, the addition of ethyl caprate and DTT are respectively 3% He of Porphyra yezoensis protease quality
15%.30 DEG C, pH 8.0, enzymolysis time 35min of hydrolysis temperature, enzyme deactivation after enzymolysis；2wt ﹪ flavor proteases are added in,
It is 6.5 to digest pH, and hydrolysis temperature is 55 DEG C, enzymolysis time 20min；
4）Isoelectric precipitation：Enzymolysis liquid is filtered, the pH value for adjusting filtrate is 4.5, and centrifuging and taking precipitates after standing；
5）Second ultrafiltration and nanofiltration：8 times of distilled water are added in the precipitation obtained in isoelectric point precipitation, are first used after being completely dissolved
Retain the film ultrafiltration of relative molecular mass 30kD, then the film ultrafiltration using retention relative molecular mass 5kD, finally by filtered solution into
Row nanofiltration desalination；
6）Macroporous resin adsorption elutes：By the liquid concentration after nanofiltration, sample concentration 0.2g/mL, loading flow velocity is 0.5mL/
Min, eluant, eluent volume are 7.8BV, and eluant, eluent is distilled water, elution flow rate 1.0mL/min；
7）Freeze-drying：The eluent of 10 ~ 30min periods is collected, is freeze-dried after its low temperature is concentrated, pre-freeze：- 40 DEG C pre-
Freeze 4h；Freeze-drying：- 35 DEG C, 2Pa freeze-dryings 6h, -7 DEG C, 15Pa is freeze-dried 2.5h；Lyophilization：12 DEG C, 55Pa liters
The dry 3.5h of China；Parsing-desiccation:12 DEG C, 4Pa parsing-desiccation 3h obtain active peptides；
8）The preparation of compound：According to 15 parts of active peptides, 7 parts of collagen small peptides, 27 parts of vitamin Cs and 4 parts of DHA mixing systems
Active peptides compound is obtained into liquor.
Routine operation in the operating procedure of the present invention is well known to those skilled in the art, herein without repeating.
Technical scheme is described in detail in embodiment described above, it should be understood that the above is only
For specific embodiments of the present invention, it is not intended to limit the invention, all any modifications made in the spirit of the present invention,
Supplement or similar fashion replacement etc., should all be included in the protection scope of the present invention.
Claims (7)
1. the purposes of active peptides compound is prepared with the deep-sea fish flesh of fish, it is characterised in that for the purposes of health products or drug.
2. the purposes according to claim 1 that active peptides compound is prepared with the deep-sea fish flesh of fish, it is characterised in that：It is described
Compound preparation method, comprise the following steps：
1）Degreasing：The deep-sea fish flesh of fish is cut into small pieces, and adds in chitosan-immobilized lipase and distilled water, constant temperature enzymolysis, enzymolysis knot
The degreasing deep-sea fish flesh of fish is obtained by filtration in enzyme deactivation after beam；
2）Deodorant：The degreasing deep-sea fish flesh of fish is dipped in 0.4 ~ 0.8h in deodorant liquid, soaking temperature is 0 ~ 4 DEG C；
3）Stepwise discretization：Minced fillet is made in deep-sea fish flesh of fish homogenate after deodorant, adds in distilled water, Porphyra yezoensis protease, the last of the ten Heavenly stems
Acetoacetic ester and DTT constant temperature digest, enzyme deactivation after enzymolysis, add flavor protease constant temperature and digest, enzyme deactivation after enzymolysis；
4）Isoelectric precipitation：Enzymolysis liquid is filtered, the pH value for adjusting filtrate is 4.0 ~ 4.5, and centrifuging and taking precipitates after standing；
5）Second ultrafiltration and nanofiltration：5 ~ 10 times of distilled water are added in the precipitation obtained in isoelectric point precipitation, after being completely dissolved first
Using the film ultrafiltration of retention 30 ~ 40kD of relative molecular mass, then the film ultrafiltration using 4 ~ 8kD of retention relative molecular mass, finally
Filtered solution is subjected to nanofiltration desalination；
6）Macroporous resin adsorption elutes：By the liquid concentration after nanofiltration, sample concentration is 0.1 ~ 0.3g/mL, and loading flow velocity is 0.4
~ 0.6mL/min, eluant, eluent volume are 7.7 ~ 7.9BV, and eluant, eluent is distilled water, and elution flow rate is 0.8 ~ 1.4mL/min；
7）Freeze-drying：The eluent of 10 ~ 30min periods is collected, is freeze-dried after its low temperature is concentrated, it is more to obtain activity
Peptide；
8）The preparation of compound：According to 10 ~ 20 parts of active peptides, 5 ~ 10 parts of collagen small peptides, 20 ~ 30 parts of vitamin Cs and 3 ~ 5
Part DHA is mixed and made into pulvis or tablet or liquor obtains active peptides compound.
3. the purposes according to claim 2 that active peptides compound is prepared with the deep-sea fish flesh of fish, it is characterised in that：It is described
Chitosan-immobilized lipase addition for 2 ~ 4 ‰, the addition of distilled water is 8 ~ 15 times of deep-sea fish flesh of fish volume, enzyme
It is 25 ~ 50 DEG C to solve temperature, and enzymolysis time is 0.8 ~ 2.5h.
4. the purposes according to claim 2 that active peptides compound is prepared with the deep-sea fish flesh of fish, it is characterised in that：It is described
Deodorant liquid ingredient and its parts by weight be 3 ~ 8 parts of tea polyphenols, 0.2 ~ 0.8 part of 10 ~ 20 parts of pure water and grape seed extract.
5. the purposes according to claim 2 that active peptides compound is prepared with the deep-sea fish flesh of fish, it is characterised in that：It is described
The addition of distilled water be 20 ~ 30 times of minced fillet volume, the addition of Porphyra yezoensis protease is 5 ~ 8wt ‰, ethyl caprate
Addition with DTT is respectively the 1 ~ 5% and 10 ~ 20% of Porphyra yezoensis protease quality, and hydrolysis temperature is 25 ~ 35 DEG C, pH 7.0
~ 9.0, enzymolysis time is 30 ~ 40min.
6. the purposes according to claim 2 that active peptides compound is prepared with the deep-sea fish flesh of fish, it is characterised in that：It is described
Flavor protease addition for 1 ~ 3wt ﹪, enzymolysis pH is 6.0 ~ 7.0, and hydrolysis temperature is 52 ~ 60 DEG C, enzymolysis time for 15 ~
25min。
7. the purposes according to claim 2 that active peptides compound is prepared with the deep-sea fish flesh of fish, it is characterised in that：It is described
Freeze-drying be：Pre-freeze：- 35 ~ -45 DEG C of 3 ~ 5h of pre-freeze；Freeze-drying：- 45 ~ -35 DEG C, 3 ~ 5Pa be freeze-dried 5 ~ 7h, -10
~ -5 DEG C, 10 ~ 20Pa is freeze-dried 2 ~ 3h；Lyophilization：10 ~ 15 DEG C, 50 ~ 60Pa lyophilizations, 3 ~ 4h；Parsing-desiccation:7~14
DEG C, 3 ~ 5Pa, 2 ~ 4h of parsing-desiccation obtain active peptides.
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| CN107245507A (en) * | 2017-06-20 | 2017-10-13 | 兰溪市捷喜食品加工技术有限公司 | Extend the preparation method of the deep-sea fish extracts in brain cell life-span |
| CN107348404A (en) * | 2017-07-01 | 2017-11-17 | 浦江县泰如食品科技有限公司 | Extend the deep-sea fish extracts in brain cell life-span |
| CN107397954A (en) * | 2017-07-31 | 2017-11-28 | 兰溪市沉默生物科技有限公司 | The purposes for the function small peptide extracted from deep-sea fish |
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| CN107245507A (en) * | 2017-06-20 | 2017-10-13 | 兰溪市捷喜食品加工技术有限公司 | Extend the preparation method of the deep-sea fish extracts in brain cell life-span |
| CN107348404A (en) * | 2017-07-01 | 2017-11-17 | 浦江县泰如食品科技有限公司 | Extend the deep-sea fish extracts in brain cell life-span |
| CN107397954A (en) * | 2017-07-31 | 2017-11-28 | 兰溪市沉默生物科技有限公司 | The purposes for the function small peptide extracted from deep-sea fish |
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| CN111493205A (en) * | 2020-04-28 | 2020-08-07 | 广东海洋大学 | Fish protein hydrolysate and preparation method thereof |
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Application publication date: 20180529 |