CN107245098A - With the tuna extract for suppressing cancer cell effect - Google Patents
With the tuna extract for suppressing cancer cell effect Download PDFInfo
- Publication number
- CN107245098A CN107245098A CN201710476258.2A CN201710476258A CN107245098A CN 107245098 A CN107245098 A CN 107245098A CN 201710476258 A CN201710476258 A CN 201710476258A CN 107245098 A CN107245098 A CN 107245098A
- Authority
- CN
- China
- Prior art keywords
- cancer cell
- freeze
- suppressing
- tuna
- tuna extract
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/461—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from fish
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Toxicology (AREA)
- Gastroenterology & Hepatology (AREA)
- Biophysics (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention discloses a kind of tuna extract for having and suppressing cancer cell effect, contain the bioactive peptide that amino acid sequence is HSHACASYYCRCLVLHPCVNCYCRCSR.Preparation process includes complex enzyme zymohydrolysis, isoelectric precipitation, ultrafiltration and nanofiltration, HPLC elutions and is freeze-dried.Have the beneficial effect that：The different peptide chain of a large amount of length is can obtain after having the restriction enzyme site of a large amount of ocean oxidized bacillus brevis production alkali proteases and undaria pinnitafida protease, enzymolysis on tuna albumen, enzymolysis efficiency is high；The purity of the bioactive peptide prepared is high, and bioactive peptide can effectively suppress untwisting for cancer cell DNA double helical structure, so as to suppress cancer cell DNA duplication, cancer cell is rested on interphase in cell division；And the secretion of E calcium mucins can be promoted, the motility of cancer cell is reduced, so as to reduce the speed of its transfer.
Description
Technical field
It is specifically a kind of that there is the tuna for suppressing cancer cell effect to extract the present invention relates to health products preparing technical field
Thing.
Background technology
It is well known that malignant tumour turns into the principal disease for threatening human health, it, which is treated, also turns into of concern
One of focus.Traditional chemotherapy and radiation also produces serious toxic side effect while treatment.The biological therapy of tumour be after
4th kind for the treatment of means of traditional remedies, current research is it has proven convenient that biological therapy is played more and more in the treatment of tumour
Big effect, and be expected to turn into the therapy for capturing tumour.
Polypeptide since earlier 1900s just attract attention, research finds that it is different from protein and is different from amino acid, has
There is medicine.Having arrived eighties of last century the nineties just has tremendous development, under the technological guidance of continuous renewal, polypeptide
Application is more and more wider, acts on 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 for being different from other drugs, just constantly affects me
Life.It is the another quantum jump and developing direction of bioscience development and polypeptide has a high potential.
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 costly.So, sight is more invested exploitation proteolysis production by people
Come in this approach of thing acquisition active peptides.Because environment and the terrestrial life that marine organisms survive are entirely different, such as high pressure, low
The extreme environment such as temperature, high temperature and high salt, in order to adapt to the marine biological environment that these are extreme, 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 amino acid sequences with bioactivity, with special protease hydrolytic, are just discharged
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 of China comes out at the top in countries in the world, but the current protide aquatic products of China
In product, in addition to a part is directly eaten, major part is made food by simple process technology and enters market, and process 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 utilized and high-valued processing.
Prior art such as Authorization Notice No. is CN103333940B Chinese invention patent, discloses one kind and utilizes hairtail system
The method of standby DPP IV peptide for inhibiting, step includes：Hairtail is added water and rubs into uniform flesh of fish slurry；Flesh of fish slurry is put into enzyme
Solve in tank, add endo protease, stirring 4~12h of hydrolysis obtains hydrolyzate；95~100 DEG C are warming up to, is kept for 10~15 minutes
Go out enzyme；Hydrolyzate is cooled to 40~60 DEG C, exoproteinase, 2~8h of stirring hydrolysis is added；95~100 DEG C are warming up to, 10 are kept
~15 minutes enzymes that go out；Endo protease is in papain, Neutrase neutral proteinases, Alcalase alkali protease
It is a kind of.Exoproteinase is Flavourzyme flavor proteases.Above-mentioned gained is gone out the hydrolyzate centrifuging and taking supernatant after enzyme；
Supernatant is added in Ultra filtration membrane device, the pressure of regulation milipore filter is 0.05~0.1MPa, the retention molecule of milipore filter
Measure as 3000Da, collection milipore filter permeate；DPP-IV peptide for inhibiting is isolated and purified using affinity chromatography method.Marine protein
The composition of matter amino acid or the sequence of amino acid are all very different with terrestrial life albumen, and the above method uses land
The protease extracted on the ground is digested to marine organisms, and enzymolysis efficiency is low.
The content of the invention
It can effectively suppress cancer cell DNA duplication it is an object of the invention to provide a kind of, cancer cell is rested on cell
Intermitotic tuna extract.
The present invention is directed to the problem of being mentioned in background technology, and the technical scheme taken is：With suppression cancer cell effect
Contain the bioactive peptide that amino acid sequence is HSHACASYYCRCLVLHPCVNCYCRCSR in tuna extract.Above-mentioned activity
Small peptide can effectively suppress untwisting for cancer cell DNA double helical structure, so as to suppress cancer cell DNA duplication, stop cancer cell
In interphase in cell division；And the secretion of E- calcium mucins can be promoted, the motility of cancer cell is reduced, so as to reduce its transfer
Speed.
With suppress cancer cell effect tuna extract preparation method, including complex enzyme zymohydrolysis, isoelectric precipitation,
Ultrafiltration and nanofiltration, HPLC elutions and freeze-drying；Complex enzyme zymohydrolysis step is to produce alkali protease using ocean oxidized bacillus brevis
Homogenate is oppressed with undaria pinnitafida protease hydrolyzed tuna, and uses ultrasound-assisted enzymolysis.Marine protein matter no matter ammonia
The composition of base acid or the sequence of amino acid are all very different with terrestrial life albumen, the alkaline egg of ocean oxidized bacillus brevis production
White enzyme and undaria pinnitafida protease are digested to tuna albumen simultaneously, can obtain a large amount of active peptide fragments.
Preferably, in complex enzyme zymohydrolysis step ocean oxidized bacillus brevis production alkali protease and undaria pinnitafida protease plus
Enter amount for 1 ~ 2wt%, hydrolysis temperature is 25 ~ 35 DEG C, pH is 7.0 ~ 9.0, enzymolysis time is 50 ~ 70min.Marine protein matter
The composition of amino acid or the sequence of amino acid are all very different with terrestrial life albumen, there is big on tuna albumen
Measure and a large amount of length differences are can obtain after the restriction enzyme site of ocean oxidized bacillus brevis production alkali protease and undaria pinnitafida protease, enzymolysis
Peptide chain, enzymolysis efficiency is high.
Preferably, the pH that isoelectric precipitation step is regulation enzymolysis liquid is 6.0 ~ 6.5, centrifuging and taking is precipitated after standing.
Under above-mentioned pH value condition, purpose bioactive peptide is in electroneutral, the solubility rapid decrease in water, while its activity is not broken
It is bad.
Dissolved preferably, ultrafiltration and nano-filtration step are the precipitation for being obtained isoelectric precipitation with pure water, first use 2-4kDa
Milipore filter pressurization ultrafiltration, then desalted with NF membrane nanofiltration, concentrate filtrate.It is further pure to purpose bioactive peptide by ultrafiltration
Change.
Preferably, elution step is 0 ~ 5min in HPLC elution steps, washed with 1 ~ 3% trifluoroacetic acid and 98 ~ 99% acetonitriles
De-, flow velocity is 0.8 ~ 1.5ml/min；5 ~ 15min, is eluted with 5 ~ 7% trifluoroacetic acids and 93 ~ 95% acetonitriles, and flow velocity is 0.8 ~
1.5ml/min；15 ~ 30min, is eluted with 8 ~ 12% trifluoroacetic acids and 88 ~ 92% acetonitriles, and flow velocity is 0.8 ~ 1.5ml/min.According to work
Property small peptide polarity design eluant component and elution action, by the bioactive peptide for having inhibitory action to cancer cell and other materials
Separate, product is further purified.
Preferably, the eluate for having absorption in 250nm is collected in elution in HPLC elution steps after terminating, merge and dense
Contracting.Bioactive peptide has stronger absorption at 250nm, can be collected according to this characteristic and deposition activity small peptide.
Preferably, freeze-drying step is：Pre-freeze：- 35 ~ -45 DEG C of 3 ~ 5h of pre-freeze；Freeze-drying：- 45 ~ -35 DEG C, 0 ~
4Pa is freeze-dried 5 ~ 7h, and -10 ~ -5 DEG C, 6 ~ 14Pa is freeze-dried 1 ~ 2h；Lyophilization：13 ~ 20 DEG C, 50 ~ 60Pa lyophilizations 2
~3h；Parsing-desiccation：5 ~ 10 DEG C, 3 ~ 5Pa, 2 ~ 4h of parsing-desiccation obtain tuna extract.Lyophilized whole process is all in program
Simple to operate under control, process is succinct, and automaticity is high, reduces human and material resources.Drying can preferably retain under low temperature
The activity of bioactive peptide；Dried under low pressure, bioactive peptide is not oxidizable, better efficacy；Bioactive peptide formation " skeleton " when freezing,
Original shape is preserved after drying, is porous loose structure and color is basically unchanged, product appearance is more excellent；Rehydration is good, freezes and lives
Property small peptide can absorb water rapidly be reduced into it is lyophilized before state, it is easy to use；Dehydration is thorough, and water content is low in bioactive peptide, typically exists
1%-3%, and be conducive to long-distance transport and long-term preservation.
Preferably, adding 40 ~ 50wt% auxiliary materials before freeze-drying, auxiliary material is mannitol.Above-mentioned auxiliary material be conducive to product into
Type, that is, prevent from shrinking when when vacuumizing, medicine disperses or is molded together with vapor.
Compared with prior art, the advantage of the invention is that：1）Marine protein matter no matter amino acid composition or
The sequence of amino acid is all very different with terrestrial life albumen, there is a large amount of ocean oxidized bacillus brevis production alkali on tuna albumen
Property protease and undaria pinnitafida protease restriction enzyme site, the different peptide chain of a large amount of length is can obtain after enzymolysis, enzymolysis efficiency is high；
2）The purity of the bioactive peptide prepared is high, and bioactive peptide can effectively suppress untwisting for cancer cell DNA double helical structure, so that
Suppress cancer cell DNA duplication, cancer cell is rested on interphase in cell division；And the secretion of E- calcium mucins can be promoted, dropped
The motility of low cancer cell, so as to reduce the speed of its transfer.
Specific embodiment
The present invention program is described further below by embodiment：
Embodiment 1：
It is containing amino acid sequence in tuna extract with suppression cancer cell effect
HSHACASYYCRCLVLHPCVNCYCRCSR bioactive peptide.Above-mentioned bioactive peptide can effectively suppress cancer cell DNA double spiral knot
Untwisting for structure, so as to suppress cancer cell DNA duplication, makes cancer cell rest on interphase in cell division；And E- calcium can be promoted to stick
The secretion of albumen, reduces the motility of cancer cell, so as to reduce the speed of its transfer.
With suppress cancer cell effect tuna extract preparation method, including complex enzyme zymohydrolysis, isoelectric precipitation,
Ultrafiltration and nanofiltration, HPLC elutions and freeze-drying；Complex enzyme zymohydrolysis step is to produce alkali protease using ocean oxidized bacillus brevis
Homogenate is oppressed with undaria pinnitafida protease hydrolyzed tuna, and uses ultrasound-assisted enzymolysis.Marine protein matter no matter ammonia
The composition of base acid or the sequence of amino acid are all very different with terrestrial life albumen, the alkaline egg of ocean oxidized bacillus brevis production
White enzyme and undaria pinnitafida protease are digested to tuna albumen simultaneously, can obtain a large amount of active peptide fragments.
The addition of ocean oxidized bacillus brevis production alkali protease and undaria pinnitafida protease is in complex enzyme zymohydrolysis step
1.5wt%, hydrolysis temperature is 30 DEG C, and pH is 8.0, and enzymolysis time is 60min.Marine protein matter no matter the composition of amino acid
Or the sequence of amino acid is all very different with terrestrial life albumen, there is a large amount of ocean oxidized bacillus brevis on tuna albumen
The different peptide chain of a large amount of length, enzymolysis efficiency are can obtain after producing the restriction enzyme site of alkali protease and undaria pinnitafida protease, enzymolysis
It is high.
Isoelectric precipitation step is that the pH of regulation enzymolysis liquid is 6.2, and centrifuging and taking is precipitated after standing.In above-mentioned pH value condition
Under, purpose bioactive peptide is in electroneutral, the solubility rapid decrease in water, while its activity is not destroyed.
Ultrafiltration and nano-filtration step are that the precipitation for being obtained isoelectric precipitation with pure water dissolves, and are first added with 2-4kDa milipore filter
Ultrafiltration is pressed, then is desalted with NF membrane nanofiltration, filtrate is concentrated.By ultrafiltration, purpose bioactive peptide is further purified.
Elution step is that elution step is 0 ~ 5min in HPLC elution steps, is eluted with 2% trifluoroacetic acid and 98% acetonitrile, stream
Speed is 1ml/min；5 ~ 15min, is eluted, flow velocity is 1ml/min with 6% trifluoroacetic acid and 94% acetonitrile；15 ~ 30min, with 10% 3
Fluoroacetic acid and the elution of 90% acetonitrile, flow velocity is 1ml/min.The eluate for having absorption in 250nm is collected, merges and concentrates.According to work
Property small peptide polarity design eluant component and elution action, by the bioactive peptide for having inhibitory action to cancer cell and other materials
Separate, product is further purified.
Elution collects the eluate for having absorption in 250nm after terminating in HPLC elution steps, merges and concentrates.Bioactive peptide
There is stronger absorption at 250nm, can be collected according to this characteristic and deposition activity small peptide.
Being freeze-dried step is：Pre-freeze：- 40 DEG C of pre-freeze 4h；Freeze-drying：- 40 DEG C, 2Pa freeze-drying 6h, -7 DEG C,
10Pa is freeze-dried 1.4h；Lyophilization：16 DEG C, 56Pa lyophilizations 2.4h；Parsing-desiccation：8 DEG C, 4Pa parsing-desiccation 3h are obtained
To tuna extract.Lyophilized whole process is all under program control, simple to operate, and process is succinct, and automaticity is high, subtracts
Human and material resources are lacked.Under low temperature drying can preferably retentive activity small peptide activity；Dried under low pressure, bioactive peptide is difficult oxygen
Change, better efficacy；Bioactive peptide formation " skeleton " when freezing, preserves original shape after drying, is porous loose structure and color is basic
Constant, product appearance is more excellent；Rehydration is good, and freeze drying activity small peptide, which can absorb water rapidly, is reduced into lyophilized preceding state, user
Just；Dehydration is thorough, and water content is low in bioactive peptide, typically in 1%-3%, and is conducive to long-distance transport and long-term preservation.
40wt% auxiliary materials are added before freeze-drying, auxiliary material is mannitol.Above-mentioned auxiliary material is conducive to formed product, that is, prevents
Shunk when medicine disperses or is molded together with vapor when vacuumizing.
Embodiment 2：
It is containing amino acid sequence in tuna extract with suppression cancer cell effect
HSHACASYYCRCLVLHPCVNCYCRCSR bioactive peptide.Above-mentioned bioactive peptide can effectively suppress cancer cell DNA double spiral knot
Untwisting for structure, so as to suppress cancer cell DNA duplication, makes cancer cell rest on interphase in cell division；And E- calcium can be promoted to stick
The secretion of albumen, reduces the motility of cancer cell, so as to reduce the speed of its transfer.
Preparation method with the tuna extract for suppressing cancer cell effect, comprises the following steps：
1）Complex enzyme zymohydrolysis：Alkali protease is produced using ocean oxidized bacillus brevis and the undaria pinnitafida protease hydrolyzed tuna flesh of fish is even
Slurries, and using 28kHZ ultrasound-assisted enzymolysis.Ocean oxidized bacillus brevis produce alkali protease and undaria pinnitafida protease plus
Enter amount for 1.9wt%, hydrolysis temperature is 32 DEG C, and pH is 7.8, and enzymolysis time is 65min；
2）Isoelectric precipitation：The pH for adjusting enzymolysis liquid with 1mol/L watery hydrochloric acid is 6.2, stands centrifuging and taking after 2h and precipitates；
3）Ultrafiltration and nanofiltration：The precipitation for being obtained isoelectric precipitation with the pure water of 2 times of volumes dissolves, and is first added with 3kDa milipore filter
Ultrafiltration is pressed, then is desalted with NF membrane nanofiltration, filtrate is concentrated；
4）HPLC is eluted：Elution step is 0 ~ 5min, is eluted with 2% trifluoroacetic acid and 98% acetonitrile, flow velocity is 1ml/min；5~
15min, is eluted, flow velocity is 1ml/min with 6% trifluoroacetic acid and 94% acetonitrile；15 ~ 30min, with 10% trifluoroacetic acid and 90% acetonitrile
Elution, flow velocity is 1ml/min.The eluate for having absorption in 250nm is collected, merges and concentrates；
5）Freeze-drying：Pre-freeze：- 40 DEG C of pre-freeze 4h；Freeze-drying：- 40 DEG C, 2Pa freeze-dryings 6h, -7 DEG C, 10Pa freezings are dry
Dry 1.4h；Lyophilization：16 DEG C, 56Pa lyophilizations 2.4h；Parsing-desiccation：8 DEG C, 4Pa parsing-desiccation 3h obtain tuna and carried
Take thing.45wt% auxiliary materials are added before freeze-drying, auxiliary material is mannitol.
Embodiment 3：
It is containing amino acid sequence in tuna extract with suppression cancer cell effect
HSHACASYYCRCLVLHPCVNCYCRCSR bioactive peptide.Above-mentioned bioactive peptide can effectively suppress cancer cell DNA double spiral knot
Untwisting for structure, so as to suppress cancer cell DNA duplication, makes cancer cell rest on interphase in cell division；And E- calcium can be promoted to stick
The secretion of albumen, reduces the motility of cancer cell, so as to reduce the speed of its transfer.
Preparation method with the tuna extract for suppressing cancer cell effect, comprises the following steps：
1）Complex enzyme zymohydrolysis：Alkali protease is produced using ocean oxidized bacillus brevis and the undaria pinnitafida protease hydrolyzed tuna flesh of fish is even
Slurries, and using 26kHZ ultrasound-assisted enzymolysis.Ocean oxidized bacillus brevis produce alkali protease and undaria pinnitafida protease plus
Enter amount for 1.8wt%, the mass ratio of ocean oxidized bacillus brevis production alkali protease and undaria pinnitafida protease is 1:2, hydrolysis temperature is
30 DEG C, pH is 8.0, and enzymolysis time is 55min；
2）Isoelectric precipitation：The pH for adjusting enzymolysis liquid with 1mol/L watery hydrochloric acid is 6.2, stands centrifuging and taking after 2h and precipitates；
3）Ultrafiltration and nanofiltration：The precipitation for being obtained isoelectric precipitation with the pure water of 2 times of volumes dissolves, and is first added with 3kDa milipore filter
Ultrafiltration is pressed, then is desalted with NF membrane nanofiltration, filtrate is concentrated；
4）HPLC is eluted：Elution step is 0 ~ 5min, is eluted with 2% trifluoroacetic acid and 98% acetonitrile, flow velocity is 1ml/min；5~
15min, is eluted, flow velocity is 1ml/min with 6% trifluoroacetic acid and 94% acetonitrile；15 ~ 30min, with 10% trifluoroacetic acid and 90% acetonitrile
Elution, flow velocity is 1ml/min.The eluate for having absorption in 250nm is collected, merges and concentrates；
5）Freeze-drying：Pre-freeze：- 40 DEG C of pre-freeze 4h；Freeze-drying：- 40 DEG C, 2Pa freeze-dryings 6h, -7 DEG C, 10Pa freezings are dry
Dry 1.4h；Lyophilization：16 DEG C, 56Pa lyophilizations 2.4h；Parsing-desiccation：8 DEG C, 4Pa parsing-desiccation 3h obtain tuna and carried
Take thing.45wt% auxiliary materials are added before freeze-drying, auxiliary material is mannitol.
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 embodiment described above, it should be understood that it is described above only
For the specific embodiment 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 be included in the scope of the protection.
SEQUENCE LISTING
<110>Lanxi City silence bio tech ltd
<120>With the tuna extract for suppressing cancer cell effect
<130> 1
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 27
<212> PRT
<213>It is artificial synthesized
<400> 1
His Ser His Ala Cys Ala Ser Tyr Tyr Cys Arg Cys Leu Val Leu His
1 5 10 15
Pro Cys Val Asn Cys Tyr Cys Arg Cys Ser Arg
20 25
Claims (9)
1. with the tuna extract for suppressing cancer cell effect, it is characterised in that：Contain ammonia in described tuna extract
Base acid sequence is HSHACASYYCRCLVLHPCVNCYCRCSR bioactive peptide.
2. according to claim 1 have the tuna extract for suppressing cancer cell effect, it is characterised in that：Described gold
The preparation method of marlin extract includes complex enzyme zymohydrolysis, isoelectric precipitation, ultrafiltration and nanofiltration, HPLC elutions and is freeze-dried.
3. according to claim 2 have the preparation method for suppressing the tuna extract that cancer cell is acted on, its feature exists
In：Described complex enzyme zymohydrolysis step is to produce alkali protease and the golden rifle of undaria pinnitafida protease hydrolyzed using ocean oxidized bacillus brevis
Fish oppresses homogenate, and uses ultrasound-assisted enzymolysis；Described ocean oxidized bacillus brevis production alkali protease and undaria pinnitafida egg
The addition of white enzyme is 1 ~ 2wt%, and hydrolysis temperature is 25 ~ 35 DEG C, and pH is 7.0 ~ 9.0, and enzymolysis time is 50 ~ 70min.
4. according to claim 2 have the preparation method for suppressing the tuna extract that cancer cell is acted on, its feature exists
In：Described isoelectric precipitation step is that the pH of regulation enzymolysis liquid is 6.0 ~ 6.5, and centrifuging and taking is precipitated after standing.
5. according to claim 2 have the preparation method for suppressing the tuna extract that cancer cell is acted on, its feature exists
In：Described ultrafiltration and nano-filtration step are that the precipitation for being obtained isoelectric precipitation with pure water dissolves, first with 2-4kDa milipore filter
Pressurization ultrafiltration, then desalted with NF membrane nanofiltration, concentrate filtrate.
6. according to claim 2 have the preparation method for suppressing the tuna extract that cancer cell is acted on, its feature exists
In：Elution step is 0 ~ 5min in described HPLC elution steps, is eluted with 1 ~ 3% trifluoroacetic acid and 98 ~ 99% acetonitriles, flow velocity is
0.8~1.5ml/min；5 ~ 15min, is eluted with 5 ~ 7% trifluoroacetic acids and 93 ~ 95% acetonitriles, and flow velocity is 0.8 ~ 1.5ml/min；15~
30min, is eluted with 8 ~ 12% trifluoroacetic acids and 88 ~ 92% acetonitriles, and flow velocity is 0.8 ~ 1.5ml/min.
7. according to claim 2 have the preparation method for suppressing the tuna extract that cancer cell is acted on, its feature exists
In：Elution collects the eluate for having absorption in 250nm after terminating in described HPLC elution steps, merges and concentrates.
8. according to claim 2 have the preparation method for suppressing the tuna extract that cancer cell is acted on, its feature exists
In：Described freeze-drying step is：Pre-freeze：- 35 ~ -45 DEG C of 3 ~ 5h of pre-freeze；Freeze-drying：- 45 ~ -35 DEG C, 0 ~ 4Pa freezings are dry
Dry 5 ~ 7h, -10 ~ -5 DEG C, 6 ~ 14Pa is freeze-dried 1 ~ 2h；Lyophilization：13 ~ 20 DEG C, 50 ~ 60Pa lyophilizations, 2 ~ 3h；Parsing
Dry：5 ~ 10 DEG C, 3 ~ 5Pa, 2 ~ 4h of parsing-desiccation obtain tuna extract.
9. according to claim 2 have the preparation method for suppressing the tuna extract that cancer cell is acted on, its feature exists
In：40 ~ 50wt% auxiliary materials are added before described freeze-drying, auxiliary material is mannitol.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710476258.2A CN107245098A (en) | 2017-06-21 | 2017-06-21 | With the tuna extract for suppressing cancer cell effect |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710476258.2A CN107245098A (en) | 2017-06-21 | 2017-06-21 | With the tuna extract for suppressing cancer cell effect |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107245098A true CN107245098A (en) | 2017-10-13 |
Family
ID=60019431
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710476258.2A Withdrawn CN107245098A (en) | 2017-06-21 | 2017-06-21 | With the tuna extract for suppressing cancer cell effect |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107245098A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107821726A (en) * | 2017-10-25 | 2018-03-23 | 浦江县欧立生物技术有限公司 | The purposes of grass carp anti-fatigue active peptide |
| CN109662263A (en) * | 2017-10-17 | 2019-04-23 | 江苏美中医疗科技有限公司 | A kind of dedicated diet of cancer dyscrasia patient |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104372056A (en) * | 2014-11-13 | 2015-02-25 | 华侨大学 | Method for preparing oxidation-resistant active substances and compound amino acids from octopus leftovers |
| CN104774899A (en) * | 2015-04-24 | 2015-07-15 | 浙江海洋学院 | Method for preparing cervical cancer resistant polypeptide from tuna dark meat protein |
-
2017
- 2017-06-21 CN CN201710476258.2A patent/CN107245098A/en not_active Withdrawn
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104372056A (en) * | 2014-11-13 | 2015-02-25 | 华侨大学 | Method for preparing oxidation-resistant active substances and compound amino acids from octopus leftovers |
| CN104774899A (en) * | 2015-04-24 | 2015-07-15 | 浙江海洋学院 | Method for preparing cervical cancer resistant polypeptide from tuna dark meat protein |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109662263A (en) * | 2017-10-17 | 2019-04-23 | 江苏美中医疗科技有限公司 | A kind of dedicated diet of cancer dyscrasia patient |
| CN107821726A (en) * | 2017-10-25 | 2018-03-23 | 浦江县欧立生物技术有限公司 | The purposes of grass carp anti-fatigue active peptide |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103992384B (en) | A kind of large yellow croaker fish bone collagen peptide and its production and use | |
| CN104774896A (en) | Preparation method for iron-chelated collagen peptide of hairtail fish-bones | |
| CN100369929C (en) | Jellyfish collagen and method for preparing the same | |
| KR20070105714A (en) | Method for preparation of capsule of seacucumber peptide | |
| CN102808010A (en) | Method for preparing antihypertensive peptides through enzymolysis of ground meat proteins of tuna | |
| Cha et al. | Screening of extracts from red algae in Jeju for potentials MarineAngiotensin-I converting enzyme (ACE) inhibitory activity | |
| KR100448672B1 (en) | A liquefied plant salt of preparation method and liquefied plant salt | |
| CN108118077A (en) | The technique that a kind of enzymatic hydrolysis salmon collagen prepares anti-oxidation peptide and antifreeze peptide | |
| CN108484723A (en) | The inhibiting peptide of tonin and its preparation method and application in Enteromorpha source | |
| CN116284341B (en) | Preparation and application of deep sea fish skin collagen peptide with low immunogenicity, blood pressure reduction and oxidation resistance | |
| CN107298710A (en) | A kind of extracting method of spirulina phycocyanin | |
| CN107245098A (en) | With the tuna extract for suppressing cancer cell effect | |
| CN103882083A (en) | Method for preparing antioxidant collagen peptide | |
| CN103819552A (en) | Cyanea capillata polypeptide growth factor, preparation method and applications thereof | |
| CN104593462A (en) | Preparation method of blue clam protein source ACE inhibition peptide | |
| CN107267583A (en) | Preparation method with the tuna extract for suppressing cancer cell effect | |
| CN110590908A (en) | Micropterus-derived antibacterial peptide additive and preparation method thereof | |
| CN107279972A (en) | Compound based on Spanish mackerel active peptides | |
| CN101130801A (en) | Preparation of active antihypertensive peptide product | |
| CN107312063A (en) | The preparation method of blunt top spirulina activated protein | |
| CN110698540B (en) | ACE inhibitory peptide derived from snakehead protein and preparation method thereof | |
| CN107245508A (en) | The hairtail activity extract repaired for Bones and joints | |
| CN107307430A (en) | Compound based on tuna extract and preparation method thereof | |
| CN107384999A (en) | From the method for Ba Sha fish-skin abstraction function protein peptides | |
| CN107312811A (en) | The preparation method for the hairtail activity extract repaired for Bones and joints |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20171013 |
|
| WW01 | Invention patent application withdrawn after publication |