CN109517868A - A kind of preparation method of the fish source collagen protein peptides of anti-aging - Google Patents
A kind of preparation method of the fish source collagen protein peptides of anti-aging Download PDFInfo
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Abstract
The invention belongs to field of biotechnology, are related to a kind of preparation method of the fish source collagen protein peptides of anti-aging；Specific steps are as follows: air bladder thaws first, cleans, drains, stripping and slicing, blends, clean, industrial alcohol is added and carries out ultrasonic wave added degreasing；After filters pressing and crushing, adds No. six solvent naphthas and carry out ultrasonic wave added degreasings；Then filters pressing is added hot water and extracts collagen, filters to take filtrate；After cooling, concentrated hydrochloric acid stomach function regulating proteinase mimic stomach cancellation ratio is added；After adjusting pH, pancreatin simulation intestines cancellation ratio is added；It finally boils, cool down, be centrifuged, obtained supernatant is subjected to ultrafiltration, filter liquor carries out nanofiltration again, collects trapped fluid and is spray-dried, obtains fish source collagen protein peptides；The usuryization that the present invention realizes fish processing waste utilizes, and the collagen peptide of preparation has the function of delaying senescence；Simple production process, the period is short, at low cost, non-environmental-pollution, and collagen peptide can be applied to the fields such as food, medicine, cosmetics.
Description
Technical field
The invention belongs to field of biotechnology, and in particular to a kind of preparation side of the fish source collagen protein peptides of anti-aging
Method.
Technical background
Collagen product is widely used to the fields such as food, medicine, cosmetics, biomaterial.Traditionally collagen egg
It is white to be mainly derived from livestock and poultry, but since people and livestock and poultry are total to the frequent hair of illness (aftosa, rabid ox disease, Streptococcus suis, bird flu)
Raw, safety increasingly allows people to worry；Meanwhile religious also makes its application and trade receive certain restrictions.China is
One fishery big country, annual a large amount of processing waste cause the serious wasting of resources and environmental pollution.Therefore, with aquatic products plus
Work waste is the collagen product that raw material production can substitute traditional livestock and poultry source, is to promote food safety and Fisheries Development
Effective way.
Skin is the maximum organ of human body, be responsible for protection, feeling, adjust body temperature, secretion, excretion and be immunized etc. many lifes
Manage function.But with advancing age, retrogression, structure atrophy, deterioration or forfeiture inevitably occur for skin, no
It can only have a negative impact to appearance and psychology, and there is close teiology to contact with many skin diseases, such as ulcer, Cuo
Sore, moss, bleb, keratosis, leucoderma, psoriasis, lupus erythematosus, cutaneum carcinoma etc..Delay skin aging process is numerous love
The unremitting pursuit of beauty scholar, and the effective way of skin disease is conquered, become one of the research hotspot of current life science.
Collagen has longer research and applicating history in delay skin aging.Early in the Eastern Han Dynasty, " typhoid fever
By " in record pig skin and have the effect of " and blood vessels, moisten skin "；Civil in China, people eat trotter, chicken feet etc. rich in collagen egg more
White food is to skin maintenance and cosmetic benefits；In current market, from pulvis, capsule to oral solution, for improving the collagen egg of skin quality
White product is numerous.In recent years, Peking University, Southern Yangtze University, Chinese Marine University, China Agricultural University and Japan, South Korea,
The effect of Germany, Italy, Brazilian researcher also confirm collagen delay skin aging from different angles respectively.
However, causing application of the collagen in delay skin aging always due to lacking sufficient experimental evidence and theoretical foundation
By dispute and query, the excessive publicity of manufacturing enterprise causes its negative event to take place frequently in addition, and confusion ring in market is raw.
Air bladder is a kind of collagen-rich fish processing waste, for a long time as food medicinal material and invigorant in
State and country in Southeast Asia gain great popularity.Traditional Chinese Medicine thinks that air bladder collagen product has improvement qi and blood, invigorates the kidney and stops nocturnal emission, supports
Face moisturizing nourishes tendon and vessel, improves a variety of physiological functions such as immune, brain tonic benefit intelligence, resisting cold clearing damp, hemostasis stasis, stimulating milk secretion detumescence, but
Its confirmation for almost not obtaining modern scientific research.Therefore, it using air bladder as raw material, produces a kind of with function of delaying senility
Collagen peptide, and its molecular mechanism is illustrated, to promotion food safety and Fisheries Development, improve people's quality of life and healthy water
It is flat to be of great significance.
Summary of the invention
In view of the deficiencies of the prior art, present invention seek to address that one of described problem；The present invention is utilized using air bladder as raw material
The technologies such as ultrasonic wave added degreasing, simulating gastrointestinal digestion, super nanofiltration separation, spray drying produce a kind of with function of delaying senility
Fish source collagen protein peptides, can be applied to the fields such as food, medicine, cosmetics.
In order to achieve the goal above, the present invention takes following technical scheme:
(1) pretreatment of raw material: after air bladder is thawed, is cleaned, is drained, stripping and slicing, blends, clean, be added industrial alcohol into
Row ultrasonic wave added degreasing；After filters pressing and crushing, adds No. six solvent naphthas and carry out ultrasonic wave added degreasings, then filters pressing is taken off
The air bladder of rouge；
(2) extraction of collagen: the degreasing air bladder that step (1) obtains is placed in hot water, extraction collagen is continuously stirred
Albumen, the filtrate obtained after filtering, as collagen solution；
(3) after the collagen solution that step (2) is obtained is cooling, concentrated hydrochloric acid stomach function regulating egg the enzymatic hydrolysis of collagen: is added
White enzyme, stirring simulation stomach cancellation ratio；After adjusting pH with sodium hydroxide again, pancreatin, stirring simulation intestines cancellation ratio is added；Finally boil
Boiling, cooling, centrifugation, gained supernatant is the enzymolysis product of collagen；
(4) separation of collagen peptide: the collagen enzymatic hydrolysis product that step (3) is obtained carries out ultrafiltration, and filter liquor is again
Then secondary carry out nanofiltration is collected trapped fluid and is spray-dried, obtains the collagen peptide with function of delaying senility.
Preferably, the ratio of air bladder described in step (1) and industrial alcohol, No. six solvent naphthas is 1kg:3~5L；It is described
The ultrasound condition of auxiliary degreasing is 10~50W/L of intensity, 3~5h of time, and 30~50 DEG C of temperature.
Preferably, hot water temperature described in step (2) is 50~60 DEG C；The degreasing air bladder and hot water ratio are 1kg:7
~10L；The mixing speed is 200~300rpm；The extraction time is 1~2h.
Preferably, temperature cooling described in step (3) is 35~40 DEG C；The concentrated hydrochloric acid and glue of the simulation stomach cancellation ratio
The volume ratio of former protein solution is 1:100~150, and pepsin is 2500~5000U/mL, and the time is 2~3h.
Preferably, adjusting pH with sodium hydroxide described in step (3) is 6.8~7.4, and pancreatin is 20000~30000U/
ML, time are 4~6h.
Preferably, the temperature of simulation stomach cancellation ratio described in step (3) and simulation intestines cancellation ratio is 37 DEG C, mixing speed
It is 150~200rpm.
Preferably, boiling time described in step (3) is 10~15min；The centrifugal condition is (5000~10000) g
× (20~30) min.
Preferably, the molecular cut off of ultrafiltration membrane described in step (4) is 1000Da, and intake pressure is 0.1~0.3MPa；
The molecular cut off of the nanofiltration membrane is 150Da, and intake pressure is 0.2~0.5MPa.
Preferably, the inlet air temperature of spray drying described in step (4) is 180~220 DEG C, and leaving air temp is 110~120
℃。
Air bladder as described in the above technical scheme includes the air bladder of Acipenseridae, gadidae, Chang section or Sciaenidae.
Beneficial effects of the present invention
1. the present invention produces collagen peptide by raw material of fish processing waste in terms of the selection of raw material, both avoided
The insecurity factor of traditional livestock and poultry collagen peptide, in turn avoids the wasting of resources and environmental pollution of air bladder, realizes fish
The higher value application of processing waste.
2. the present invention utilizes the side of ultrasonic wave added industrial alcohol and No. six solvent naphtha degreasings in terms of the pretreatment of raw material
Method, effect is good, and low energy consumption, the time is short, is easily achieved industrialization.
3. the present invention uses Hot water extraction, and collagen recovery rate is higher, molecule knot in terms of the extraction of collagen
Structure is also more complete, and compared to traditional salt, acid, Enzymatic Extraction, simple process, the period is short, at low cost, and non-environmental-pollution.
4. in terms of the enzymatic hydrolysis of collagen, the method that the present invention is digested by in-vitro simulated human gastrointestinal tract, not only greatly
The digestibility of collagen peptide is improved greatly, and effectively prevents being orally ingested rear human gastrointestinal tract and re-digests enzymatic hydrolysis and cause
Bioactivity significantly reduce, the shortcomings that even disappearing completely.
5. in terms of the separation of collagen peptide, the method that the present invention utilizes UF membrane (ultrafiltration, nanofiltration) is had concurrently dense
The function of contracting, separation, purifying, and have efficient, energy-saving and environmental protection, the advantage that process is simple, easily controllable, it both remains enzymatic hydrolysis and produces
The effect of object component, and improve the biological value of final product.
6. the present invention passes through rat Long-term Feeding experiment card in terms of the biological safety of collagen peptide and physiological activity
It is real, the collagen peptide not only biological safety with higher, but also can significantly improve skin texture and matrix is flat
Weighing apparatus improves antioxidase activity, alleviates oxidative stress, effective delay skin aging process；Fibroblasts in vitro culture experiment
It has also been found that collagen peptide (Hyp-Gly) can not only promote cell Proliferation, additionally it is possible to pass through p38 protein phosphorylation, activation
MAPK intracellular signal transduction pathway, regulate and control I-type collagen and its mRNA expression, effectively maintain skin normal organization and
Biochemical functions.
Detailed description of the invention
Fig. 1 is the SDS- polyacrylamide gel electrophoresis map of collagen prepared by embodiment 1.
Fig. 2 is the gel permeation chromatogram spectrum of collagen peptide prepared by embodiment 1.
Fig. 3 is influence of the collagen peptide of the preparation of embodiment 1 to rat skin institutional framework；Wherein, HE: hematoxylin-
Eosin stains；MS:Masson dyeing；SRO: the sirius red stains under normal light；SRP: the sirius red stains under polarisation；
BC: blank control group；YC: young control；CT: collagen peptide processing group.
A is influence of the collagen peptide of different molecular structures to L929 cell Proliferation in Fig. 4；B is various concentration
Influence of the Hyp-Gly to L929 cell Proliferation, wherein bFGF is basic fibroblast growth factor；" * " is indicated and control group
There are significant difference (P < 0.05).
Fig. 5 is the influence that Hyp-Gly secretes L929 cell I-type collagen and COL1A1 mRNA is expressed；Wherein, " * "
Indicate that there are significant difference (P < 0.05) with control group.
Fig. 6 is the L929 cell COL1A1 mRNA expression that intracellular signal transduction pathway inhibitor induces Hyp-Gly
Influence；Wherein, Genistein is genistein, and H7 is protein kinase C pathway inhibitor；" * " indicates exist with control group
Significant difference (P < 0.05).
Fig. 7 is influence of the Hyp-Gly to L929 cell p38 protein phosphorylation level；Wherein, " * " expression is deposited with control group
In significant difference (P < 0.05).
Specific embodiment
In conjunction with following instance, the present invention will be described in further detail.
Embodiment 1:
10kg sturgeon fish glue is thawed, cleaning drains, stripping and slicing (3 × 3cm), and 5 meshes are crossed after blending and remove impurity；Then, add
Enter 50L industrial alcohol, degreasing 3h is assisted with 50W/L ultrasonic wave at 30 DEG C；After filters pressing and crushing, No. six solvents of 30L are added
Oil assists degreasing 5h at 50 DEG C with 10W/L ultrasonic wave；Then filters pressing obtains the sturgeon fish glue about 3.0kg of degreasing；
50 DEG C of hot water 30L is added into above-mentioned degreasing air bladder, extracts collagen 1h under 200rpm stirring；Then it crosses
20 meshes are cooled to 35 DEG C, and concentrated hydrochloric acid 0.3L and pepsin are added into filtrate to 2500U/mL, continuously stirs in 150rpm
3h simulates stomach cancellation ratio；PH to 6.8 is adjusted with NaOH again, pancreatin is added to 20000U/mL, continuously stirs 6h simulation in 200rpm
Intestines cancellation ratio；10min is finally boiled, cooling, 10000g is centrifuged 20min, and gained supernatant is collagen enzymatic hydrolysis product；
Using molecular cut off be 1000Da ultrafiltration membrane, at intake pressure 0.1MPa to collagen digest product into
Row ultrafiltration collects filter liquor up to no solute filters out；Recycling molecular cut off is the nanofiltration membrane of 150Da, in intake pressure
Nanofiltration is carried out to above-mentioned filter liquor under 0.2MPa, until filtering out without solute, collects trapped fluid in 180 DEG C of inlet air temperature, outlet air
It is spray-dried under the conditions of 110 DEG C of temperature, white powder is obtained, as with the collagen peptide of delaying senility function.
Embodiment 2:
10kg gadus fish glue is taken to thaw, cleaning drains, stripping and slicing (about 1 × 1cm), and 10 meshes are crossed after blending and remove impurity；It is added
30L industrial alcohol assists degreasing 5h at 50 DEG C with 10W/L ultrasonic wave；After filters pressing and crushing, No. six solvent naphthas of 50L are added,
Degreasing 3h is assisted with 50W/L ultrasonic wave at 30 DEG C；Last filters pressing obtains the gadus fish glue about 3.5kg of degreasing；
60 DEG C of hot water 25L are added into above-mentioned degreasing air bladder, extract collagen 2h under 300rpm stirring；Then 50 are crossed
Mesh is cooled to 40 DEG C, and concentrated hydrochloric acid 0.2L and pepsin are added into filtrate to 5000U/mL, continuously stirs 2h in 200rpm
Simulate stomach cancellation ratio；PH to 7.4 is adjusted with NaOH again, pancreatin is added to 30000U/mL, continuously stirs 4h simulation intestines in 150rpm
Cancellation ratio；15min is finally boiled, cooling, 5000g is centrifuged 30min, and gained supernatant is collagen enzymatic hydrolysis product；
Using molecular cut off be 1000Da ultrafiltration membrane, at intake pressure 0.3MPa to collagen digest product into
Row ultrafiltration collects filter liquor until filtering out almost without solute；Recycling molecular cut off is the nanofiltration membrane of 150Da, in influent pressure
Nanofiltration is carried out to above-mentioned filter liquor under power 0.5MPa, until filtering out almost without solute, collects trapped fluid in inlet air temperature 220
DEG C, be spray-dried under the conditions of 120 DEG C of leaving air temp, white powder is obtained, as with the collagen egg of delaying senility function
White peptide.
Embodiment 3:
10kg Channel-catfish air bladder is thawed, cleaning drains, stripping and slicing (about 2 × 2cm), and 5 meshes are crossed after blending and remove impurity；It is added
40L industrial alcohol assists degreasing 4h at 40 DEG C with 30W/L ultrasonic wave；After filters pressing and crushing, No. six solvent naphthas of 40L are added,
Degreasing 4h is assisted with 30W/L ultrasonic wave at 40 DEG C；Last filters pressing obtains degreasing Channel-catfish air bladder about 3.2kg；
55 DEG C of hot water 27.5L are added into above-mentioned pretreated air bladder, extract collagen under the stirring of 250rpm
1.5h；Then 30 meshes are crossed, are cooled to 37 DEG C, concentrated hydrochloric acid 0.2L and pepsin are added into filtrate to 4000U/mL,
2.5h simulation stomach cancellation ratio is continuously stirred under 175rpm；PH to 7.0 is adjusted with NaOH again, pancreatin is added to 25000U/mL,
5h simulation intestines cancellation ratio is continuously stirred under 175rpm；12min is finally boiled, cooling, 7500g is centrifuged 25min, and gained is colorless and transparent
Supernatant be collagen enzymatic hydrolysis product；
Using molecular cut off be 1000Da ultrafiltration membrane, at intake pressure 0.2MPa to collagen digest product into
Row ultrafiltration collects filter liquor until filtering out almost without solute；Recycling molecular cut off is the nanofiltration membrane of 150Da, in influent pressure
Power 0.3MPa to above-mentioned filter liquor carry out nanofiltration, to being filtered out almost without solute until, collection trapped fluid 200 DEG C of inlet air temperature,
It is spray-dried under the conditions of 115 DEG C of leaving air temp, white powder is obtained, as with the collagen of delaying senility function
Peptide.
Performance test
The collagen peptide that selection example 1 obtains carries out subsequent performance test.
Test method:
(1) analysis of physical and chemical feature of collagen and polypeptide
SDS- polyacrylamide gel electrophoresis (PAGE): using 5% separation gel and 3% concentration glue discontinuous electro-phoresis system；
Amino acid composition: automatic using amino acid after sample is hydrolyzed for 24 hours in 110 DEG C of the 6M hydrochloric acid containing 1% (v/v) phenol
Analyzer detection；True digestibility: rat metabolism cage method；Molecular weight distribution: gel permeation chromatography (GPC) method, using Waters
120 column of Ultrahydrogel (7.8 × 300mm), distilled water elution speed are 0.5mL/min, Detection wavelength 220nm.
(2) the delay skin aging evaluation of effect of collagen peptide
30 6 monthly age of Healthy female SD rats are randomly divided into 2 groups: blank control group (BC, AIN-93M standard feed) and
Collagen peptide processing group (CT, 4% collagen peptide+96%AIN-93M standard feed).Ad lib and drinking-water 12 months
Afterwards, dislocation is put to death, and collects Rats Organs and Tissues and skin of back.Using 3 monthly age of Healthy female SD rat as young control (YC,
AIN-93M standard feed).
Hydroxyproline content: colorimetric method for determining is carried out according to ISO3496；Oxidative stress: illustrate to survey according to kit
Determine catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) vigor and malonaldehyde
(MDA) content；Institutional framework: after skin is fixed, is dehydrated, is embedded, is sliced, hematoxylin-eosin (HE), Masson are carried out respectively
And sirius red stains, then dermis thickness, collagenous fibres density and III, type i collagen are analyzed using software Halcon 12.0
Protein ratio.
(3) Study on Molecular Mechanism of collagen peptide delay skin aging
Fibroblast (L929) is recovered, secondary culture is inoculated in 96 porocyte culture plates after adjusting cell concentration, adds
Enter the FBS-DMEM culture medium containing collagen peptide: blood counting chamber carries out cell count；ELISA detects I-type collagen
Expression quantity；RT-PCR measures COL1A1 mRNA expression；Blocked method analyzes intracellular signal transduction pathway；Western
Blotting detects signal effect molecule.
Test result:
(1) physicochemical property of collagen and polypeptide；
Using 50 DEG C of hot water extraction 1h, collagen recovery rate is up to 91.3%；Pass through simulating gastrointestinal digestion and super nanofiltration
The collagen peptide yield of separation method preparation is 66.5%, true digestibility 98.6%.
Fig. 1 is the SDS-PAGE map of collagen.The collagen has relatively complete molecular structure, includes at least
Two α chains (α 1 and α 2) and its dimer (β) and polymer, are presented typical I-type collagen electrophorogram feature.
Fig. 2 is the gpc chromatogram of collagen peptide.The collagen peptide molecular weight is concentrated mainly on 200-600Da, average mark
Son amount is 374.2Da；Amino acid composition analysis shows (table 1), which is rich in glycine (accounting for about 1/3), alanine,
Proline, glutamic acid and hydroxyproline content are relatively high, and tyrosine and histidine content are relatively low, be not detected tryptophan and
Cysteine.
The amino acid of 1 collagen peptide of table forms (residue number/1000 residue)
(2) the delay skin aging effect of collagen peptide；
From table 2 it can be seen that weight, food-intake, organ index and the blank control of the collagen peptide processing group rat
Group is compared to without significant difference, it was confirmed that its higher biological safety.
Fig. 3 is influence of the collagen peptide to rat skin institutional framework.HE dyeing shows blank control group rat skin
Skin has the characteristic feature of the agings such as sparse cell, fiber disorder, true epidermis blur boundary, and dermis thickness also compares young control
Reduce 20.1%.Collagen peptide processing group rat skin is more orderly, compact and complete, dermis thickness and young control
Also without significant difference.Observe that blank control group rat skin collagenous fibres are loose, unordered and fragmentation by Masson dyeing,
And collagen peptide processing group collagenous fibres are closely orderly, density increases 22.2%, and hydroxyproline increases 27.7%.Its wolf
Star red colouring also shows that young control rat skin is mainly orange-yellow or shiny red I-type collagen, blank control group
Predominantly green type III, and collagen peptide processing significantly reduces the ratio of type III and I-type collagen.The result table
Bright collagen peptide can significantly improve the institutional framework and extracellular matrix dynamic equilibrium of rat skin.
Influence of 2 collagen peptide of table to rat physical signs and skin characteristic
In addition, CAT, SOD, GSH-Px vigor of collagen peptide processing group rat skin are respectively increased compared with blank control group
48.0%, 49.4% and 24.2%, MDA content then reduces 31.5%.This shows that collagen peptide improves antioxidase
Vigor, it is suppressed that lipid peroxidation alleviates the oxidative stress of skin.
(3) molecular mechanism of collagen peptide delay skin aging；
Fig. 4 is the collagen peptide (a) of different molecular structures and the Hyp-Gly (b) of various concentration to L929 cell Proliferation
Influence；It can be seen that from Fig. 4 a after handling L929 cell 7d using the collagen peptide of different molecular structures, Pro-Hyp,
Hyp-Gly, Ala-Hyp, Ala-Hyp-Gly and Leu-Hyp significantly promote cell Proliferation, and wherein Hyp-Gly promotes cell
Proliferative capacity is most strong, and cell number improves 2.09 times than control group, and is in dose-dependant in 0~200 μm of ol concentration range
Relationship (Fig. 4 b).
Fig. 5 is the influence that Hyp-Gly secretes L929 cell I-type collagen and COL1A1 mRNA is expressed；Utilize Hyp-
After Gly handles L929 cell for 24 hours, the content of I-type collagen improves 3.74 times than control group, 1 chain gene COL1A1 of α
The expression quantity of mRNA also increases 2.46 times.
Fig. 6 is shadow of the cell-signaling pathways inhibitor to the Hyp-Gly L929 cell COL1A1 mRNA expression induced
It rings；Genistein and H7 have blocked the signal path of Hyp-Gly, significantly reduce the expression of COL1A1 mRNA.
Fig. 7 is influence of the Hyp-Gly to L929 cell p38 protein phosphorylation level；Hyp-Gly is in 30min and 60min
Significantly improve p38 phosphorylation level；This shows that Hyp-Gly passes through induction p38 phosphorylation activation MAPK signal path, in turn
Promote COL1A1 mRNA expression.
In summary: (1) using the collagen recovery rate with higher (91.3%) of 50 DEG C of hot water extractions and relatively
Complete I-type collagen molecular structure；(2) collagen peptide prepared by simulating gastrointestinal digestion and super nanofiltration separation method
Yield is 66.5%, true digestibility 98.6%, and molecular weight is concentrated mainly on 200-600Da, average molecular weight 374.2Da,
Rich in glycine, alanine, proline, glutamic acid and hydroxyproline etc.；(3) collagen peptide bio-safety with higher
Property, can improve the institutional framework and matrix equilibrium of skin, alleviate skin oxidative stress, delay skin aging process；(4) glue
Former protein peptides main component can not only promote fibroblast proliferation, and by induction p38 protein phosphorylation, activate MAPK
Intracellular signal transduction pathway promotes COL1A1 mRNA expression and I-type collagen synthesis secretion.
Illustrate: above embodiments are only to illustrate the present invention and not limit the technical scheme described by the invention；Therefore,
Although this specification is referring to above-mentioned each embodiment, the present invention has been described in detail, the common skill of this field
Art personnel should be appreciated that and still can modify to the present invention or equivalent replacement；And all do not depart from spirit of the invention and
The technical solution and its improvement of range, should all cover in scope of the presently claimed invention.
Claims (9)
1. a kind of preparation method of the fish source collagen protein peptides of anti-aging, which is characterized in that steps are as follows:
(1) after air bladder is thawed, is cleaned, is drained, stripping and slicing, blends, clean, it is auxiliary that industrial alcohol ultrasound the pretreatment of raw material: is added
Help degreasing；After filters pressing and crushing, No. six solvent naphtha ultrasonic wave added degreasings are added, then filters pressing obtains degreasing air bladder；
(2) extraction of collagen: the degreasing air bladder that step (1) obtains being placed in hot water, extraction collagen is continuously stirred,
Filtrate, as collagen solution are obtained after filtering；
(3) enzymatic hydrolysis of collagen: after the collagen solution that step (2) is obtained is cooling, being added concentrated hydrochloric acid and pepsin,
Stirring simulation stomach cancellation ratio；After adjusting pH with sodium hydroxide again, pancreatin, stirring simulation intestines cancellation ratio is added；It finally boils, is cold
But, it is centrifuged, gained supernatant is the enzymolysis product of collagen；
(4) separation of collagen peptide: the collagen enzymatic hydrolysis product that step (3) is obtained carries out ultrafiltration, filter liquor again into
Row nanofiltration is collected trapped fluid and is spray-dried, obtains the collagen peptide with function of delaying senility.
2. a kind of preparation method of the fish source collagen protein peptides of anti-aging according to claim 1, which is characterized in that step
Suddenly the ratio of air bladder described in (1) and industrial alcohol, No. six solvent naphthas is 1kg:3 ~ 5L；The ultrasound condition of the auxiliary degreasing
It is 10 ~ 50W/L of intensity, 3 ~ 5h of time, 30 ~ 50 DEG C of temperature.
3. a kind of preparation method of the fish source collagen protein peptides of anti-aging according to claim 1, which is characterized in that step
Suddenly hot water temperature described in (2) is 50 ~ 60 DEG C；The degreasing air bladder and hot water ratio are 1kg:7 ~ 10L；The mixing speed is
200~300rpm；The extraction time is 1 ~ 2h.
4. a kind of preparation method of the fish source collagen protein peptides of anti-aging according to claim 1, which is characterized in that step
Suddenly cooling temperature described in (3) is 35 ~ 40 DEG C；The volume ratio of the concentrated hydrochloric acid and collagen solution is 1:100 ~ 150；It is described
Pepsin is 2500 ~ 5000U/mL；The time of the stirring simulation stomach cancellation ratio is 2 ~ 3h.
5. a kind of preparation method of the fish source collagen protein peptides of anti-aging according to claim 1, which is characterized in that step
Suddenly adjusting pH with sodium hydroxide described in (3) is 6.8 ~ 7.4；The pancreatin is 20000 ~ 30000U/mL；Intestines are simulated in the stirring
The time of cancellation ratio is 4 ~ 6h.
6. a kind of preparation method of the fish source collagen protein peptides of anti-aging according to claim 1, which is characterized in that step
Suddenly the temperature of simulation stomach cancellation ratio and intestines cancellation ratio described in (3) is 37 DEG C, and mixing speed is 150 ~ 200rpm.
7. a kind of preparation method of the fish source collagen protein peptides of anti-aging according to claim 1, which is characterized in that step
Suddenly boiling time described in (3) is 10 ~ 15min；The centrifugal condition is (5000 ~ 10000) g × (20 ~ 30) min.
8. a kind of preparation method of the fish source collagen protein peptides of anti-aging according to claim 1, which is characterized in that step
The molecular cut off of ultrafiltration membrane is 1000Da when carrying out ultrafiltration described in (4) suddenly, and intake pressure is 0.1 ~ 0.3MPa；The progress
The molecular cut off of nanofiltration membrane is 150Da when nanofiltration, and intake pressure is 0.2 ~ 0.5MPa.
9. a kind of preparation method of the fish source collagen protein peptides of anti-aging according to claim 1, which is characterized in that step
Suddenly the inlet air temperature of spray drying described in (4) is 180 ~ 220 DEG C, and leaving air temp is 110 ~ 120 DEG C.
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| CN110577975A (en) * | 2019-09-24 | 2019-12-17 | 福州海锐黎思生物科技有限责任公司 | Method for extracting and preparing swimming bladder collagen oligopeptide |
| CN110713534A (en) * | 2019-11-29 | 2020-01-21 | 福建农林大学 | Collagen peptide with photoaging improvement effect and preparation method thereof |
| CN110973636A (en) * | 2019-11-19 | 2020-04-10 | 杭州民生健康药业有限公司 | Composition containing vitamin D, preparation method thereof and application thereof in increasing bone density |
| CN111635920A (en) * | 2020-05-21 | 2020-09-08 | 内蒙古元本生物医药科技有限公司 | Method for preparing donkey bone collagen peptide powder by two-stage bionic enzymatic hydrolysis technology |
| CN112409933A (en) * | 2020-10-21 | 2021-02-26 | 江苏大学 | Production method of fish-derived gelatin and nanofiber membrane |
| CN113698471A (en) * | 2021-07-04 | 2021-11-26 | 海南欣芯生物科技有限公司 | Pickling unit of PH value regulation of being convenient for is used in collagen production |
| CN115191596A (en) * | 2022-07-08 | 2022-10-18 | 江苏大学 | Lactobacillus plantarum-based collagen peptide product for delaying senescence and production method thereof |
| CN116179634A (en) * | 2023-04-27 | 2023-05-30 | 北京盛美诺生物技术有限公司 | III type collagen peptide and preparation method thereof |
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110577975A (en) * | 2019-09-24 | 2019-12-17 | 福州海锐黎思生物科技有限责任公司 | Method for extracting and preparing swimming bladder collagen oligopeptide |
| CN110973636A (en) * | 2019-11-19 | 2020-04-10 | 杭州民生健康药业有限公司 | Composition containing vitamin D, preparation method thereof and application thereof in increasing bone density |
| CN110713534A (en) * | 2019-11-29 | 2020-01-21 | 福建农林大学 | Collagen peptide with photoaging improvement effect and preparation method thereof |
| CN111635920A (en) * | 2020-05-21 | 2020-09-08 | 内蒙古元本生物医药科技有限公司 | Method for preparing donkey bone collagen peptide powder by two-stage bionic enzymatic hydrolysis technology |
| CN112409933A (en) * | 2020-10-21 | 2021-02-26 | 江苏大学 | Production method of fish-derived gelatin and nanofiber membrane |
| CN113698471A (en) * | 2021-07-04 | 2021-11-26 | 海南欣芯生物科技有限公司 | Pickling unit of PH value regulation of being convenient for is used in collagen production |
| CN113698471B (en) * | 2021-07-04 | 2023-03-17 | 海南欣芯生物科技有限公司 | Pickling device convenient for PH value adjustment for collagen production |
| CN115191596A (en) * | 2022-07-08 | 2022-10-18 | 江苏大学 | Lactobacillus plantarum-based collagen peptide product for delaying senescence and production method thereof |
| CN115191596B (en) * | 2022-07-08 | 2023-10-20 | 江苏大学 | Lactobacillus plantarum-based anti-aging collagen peptide product and production method thereof |
| CN116179634A (en) * | 2023-04-27 | 2023-05-30 | 北京盛美诺生物技术有限公司 | III type collagen peptide and preparation method thereof |
| CN116179634B (en) * | 2023-04-27 | 2023-08-18 | 北京盛美诺生物技术有限公司 | III type collagen peptide and preparation method thereof |
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