CN107987121A - Suppress the preparation method of the polypeptide extract of cancer cell - Google Patents
Suppress the preparation method of the polypeptide extract of cancer cell Download PDFInfo
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- CN107987121A CN107987121A CN201711092034.8A CN201711092034A CN107987121A CN 107987121 A CN107987121 A CN 107987121A CN 201711092034 A CN201711092034 A CN 201711092034A CN 107987121 A CN107987121 A CN 107987121A
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- cancer cell
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/16—Extraction; Separation; Purification by chromatography
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- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- Medicinal Chemistry (AREA)
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- Engineering & Computer Science (AREA)
- Water Supply & Treatment (AREA)
- Peptides Or Proteins (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses the preparation method for the polypeptide extract for suppressing cancer cell, method includes the pretreatment of Optimization of Low Value Fish, the preparation of albumen, the decomposition of albumen and isolates and purifies.Have the beneficial effect that：Addition of this method without catalyst or enzyme, avoids separating step；Brine boiling Optimization of Low Value Fish, plays Optimization of Low Value Fish the effect of sterilization, and the protein in Optimization of Low Value Fish can be moderately denatured at this temperature, and good condition is provided for follow-up autoclaving；Gained polypeptide can effectively suppress the propagation of cancer cell.
Description
Technical field
The present invention relates to technical field of biological product preparation, more particularly, to the preparation for the polypeptide extract for suppressing cancer cell
Method.
Background technology
Due to the popularization of modern Fishing technology, larger fish resource is gradually destroyed, so that small fish in biological chain,
Shrimp breeds rapidly.In addition, the small fish, shrimp and disposal from fishery product processing in catches typically constitute from the 28% of catches.By
In the further investigation for lacking process and utilization technology, low value fish are mainly for the production of the low value-added product such as fish meal at present, even
Directly abandoned as discarded object, not only waste of resource, but also also result in the pollution of ocean, terrestrial environment.If can be by Optimization of Low Value Fish
Further develop and utilize, the added value of Optimization of Low Value Fish and its accessory substance will be greatly improved, to make full use of this marine resources to open
Ward off new approach.
The content of the invention
It is an object of the invention to provide the preparation method for the polypeptide extract for suppressing cancer cell, this method without catalyst or
The addition of enzyme, avoids separating step, and the high income of gained polypeptide, can effectively suppress the propagation of cancer cell.
The present invention is directed to the problem of being mentioned in background technology, and the technical solution taken is：Suppress the polypeptide extraction of cancer cell
The preparation method of thing, method comprise the following steps：
The pretreatment of Optimization of Low Value Fish：By Optimization of Low Value Fish boiling in the saline solution that mass fraction is 22%-25%, boiling temperature 81-
87 DEG C, time 17-21min, is cooled to room temperature, wherein, the mass ratio of Optimization of Low Value Fish and saline solution is 1/1.2-1.5, pretreatment
Process avoids the influence of impurity, while with brine boiling Optimization of Low Value Fish, the effect of sterilization is played to Optimization of Low Value Fish, and at this temperature
Protein in Optimization of Low Value Fish can be moderately denatured, and good condition is provided for follow-up autoclaving；
The preparation of albumen：Optimization of Low Value Fish is put into pressure cooker, the boiling at absolute pressure 0.15-0.2MPa, 121-126 DEG C of temperature
0.6-1h, obtains protein liquid, and under the conditions of being somebody's turn to do, the albumen dissolution rate contained by Optimization of Low Value Fish is fast, and dissolution rate is high, obtains low value fish resource
Make full use of, substantially increase the economic value of method；
The decomposition of albumen：Gained protein liquid is continued into boiling 3.1- at absolute pressure 0.1-0.15MPa, 116-122 DEG C of temperature
3.8h, obtains protein breakdown liquid, and the addition without catalyst during protein breakdown, avoids the separation process of subsequent catalyst, letter
Change processing step, and avoided the loss of Optimization of Low Value Fish Protein in separation process, reduce processing cost；
Isolate and purify：Protein breakdown liquid is filtered through the ultrafiltration membrane that molecular cut off is 1kDa, trapped fluid is collected, by trapped fluid
It is added to according to volume ratio in the chromatographic column equipped with 8-10 times of D101 macroreticular resin, impurity is removed with 3-5 times of column volume water elution,
Then it is that 58%-61% ethanol waters are eluted with the mass fraction of 5-8 times of column volume, ethanol eluate is removed in 50 DEG C of revolvings
Remove ethanol, be freeze-dried to obtain active peptides.The process is easy to operate, reproducible, and Elution range is wide, can be according to actual conditions
The condition of being amplified realizes industrialized production.
Preferably, during isolating and purifying, account for ethanol in the preceding addition into ethanol eluate of ethanol eluate revolving and wash
The diphenyl-bromomethane and 0.03% 3- cresotinic acids of de- liquid mass fraction 0.02%.The diphenyl-bromomethane that is added, 3- first
Base salicylic acid has synergistic effect with ethanol, improves the yield of active peptides during revolving.
Compared with prior art, the advantage of the invention is that：The polypeptide extract disclosed by the invention for suppressing cancer cell
Preparation method includes the pretreatment of Optimization of Low Value Fish, the preparation of albumen, the decomposition of albumen and isolates and purifies, this method without catalyst or
The addition of enzyme, avoids separating step；Brine boiling Optimization of Low Value Fish, plays Optimization of Low Value Fish the effect of sterilization, and low at this temperature
Protein in value fish can be moderately denatured, and good condition is provided for follow-up autoclaving.
Embodiment
Below by embodiment, the present invention will be further described：
Embodiment 1：
Suppress the preparation method of the polypeptide extract of cancer cell, the described method comprises the following steps：
1) pretreatment of Optimization of Low Value Fish：By Optimization of Low Value Fish boiling in the saline solution that mass fraction is 22%, boiling temperature is 81 DEG C, when
Between be 17min, be cooled to room temperature, wherein, the mass ratio of Optimization of Low Value Fish and saline solution is 1/1.2, and preprocessing process avoids impurity
Influence, while with brine boiling Optimization of Low Value Fish, the effect of sterilization, and the albumen in Optimization of Low Value Fish at this temperature are played to Optimization of Low Value Fish
Matter can be moderately denatured, and good condition is provided for follow-up autoclaving；
2) preparation of albumen：Optimization of Low Value Fish is put into pressure cooker, the boiling 0.6h at absolute pressure 0.15MPa, 121 DEG C of temperature, obtains
Protein liquid, under the conditions of being somebody's turn to do, the albumen dissolution rate contained by Optimization of Low Value Fish is fast, and dissolution rate is high, low value fish resource has been obtained abundant profit
With substantially increasing the economic value of method；
3) decomposition of albumen：Gained protein liquid is continued into boiling 3.1h at absolute pressure 0.1MPa, 116 DEG C of temperature, obtains albumen
Decomposed solution, the addition without catalyst during protein breakdown, avoids the separation process of subsequent catalyst, simplifies technique step
Suddenly, and the loss of Optimization of Low Value Fish Protein in separation process is avoided, reduces processing cost；
4) isolate and purify：Protein breakdown liquid is filtered through the ultrafiltration membrane that molecular cut off is 1kDa, is collected trapped fluid, will be retained
Liquid is added in the chromatographic column equipped with 8 times of D101 macroreticular resins according to volume ratio, removes impurity with 3 times of column volume water elutions, so
Eluted afterwards with the mass fractions of 5 times of column volumes for 58% ethanol water, ethanol eluate 50 DEG C of revolvings remove ethanol,
It is freeze-dried to obtain active peptides.The process is easy to operate, reproducible, and Elution range is wide, can be put according to actual conditions
Big condition realizes industrialized production.
Embodiment 2：
Suppress the preparation method of the polypeptide extract of cancer cell, the described method comprises the following steps：It is the pre- place of Optimization of Low Value Fish first
Reason：By Optimization of Low Value Fish boiling in the saline solution that mass fraction is 25%, boiling temperature is 87 DEG C, and time 21min, is cooled to room
Temperature, wherein, the mass ratio of Optimization of Low Value Fish and saline solution is 1/1.5；Followed by the preparation of albumen：Optimization of Low Value Fish is put into pressure cooker,
Absolute pressure 0.2MPa, boiling 1h at 126 DEG C of temperature, obtains protein liquid, should under the conditions of, the albumen dissolution rate contained by Optimization of Low Value Fish
It hurry up, dissolution rate is high, low value fish resource is fully used, substantially increases the economic value of method；Followed by albumen
Decomposition：Gained protein liquid is continued into boiling 3.8h at absolute pressure 0.15MPa, 122 DEG C of temperature, obtains protein breakdown liquid；Most
After be to isolate and purify：Protein breakdown liquid is filtered through the ultrafiltration membrane that molecular cut off is 1kDa, trapped fluid is collected, by trapped fluid
It is added to according to volume ratio in the chromatographic column equipped with 10 times of D101 macroreticular resins, removes impurity with 5 times of column volume water elutions, then
Eluted with the mass fractions of 8 times of column volumes for 61% ethanol water, ethanol eluate removes ethanol, cold in 50 DEG C of revolvings
Lyophilized dry active peptides.The process is easy to operate, reproducible, and Elution range is wide, can be amplified according to actual conditions
Condition realizes industrialized production.
Embodiment 3：
Suppress the preparation method of the polypeptide extract of cancer cell, the described method comprises the following steps：It is the pre- place of Optimization of Low Value Fish first
Reason：By Optimization of Low Value Fish boiling in the saline solution that mass fraction is 24%, boiling temperature is 86 DEG C, and time 21min, is cooled to room
Temperature, wherein, the mass ratio of Optimization of Low Value Fish and saline solution is 1/1.4；Followed by the preparation of albumen：Optimization of Low Value Fish is put into pressure cooker,
Absolute pressure 0.2MPa, boiling 1h at 125 DEG C of temperature, obtains protein liquid, should under the conditions of, the albumen dissolution rate contained by Optimization of Low Value Fish
It hurry up, dissolution rate is high, low value fish resource is fully used, substantially increases the economic value of method；Followed by albumen
Decomposition：Gained protein liquid is continued into boiling 3.8h at absolute pressure 0.15MPa, 121 DEG C of temperature, obtains protein breakdown liquid；Most
After be to isolate and purify：Protein breakdown liquid is filtered through the ultrafiltration membrane that molecular cut off is 1kDa, trapped fluid is collected, by trapped fluid
It is added to according to volume ratio in the chromatographic column equipped with 10 times of D101 macroreticular resins, removes impurity with 5 times of column volume water elutions, then
Eluted with the mass fraction of 8 times of column volumes for 60% ethanol water, added into gained ethanol eluate and account for ethanol elution
The diphenyl-bromomethane of liquid mass fraction 0.02% and 0.03% 3- cresotinic acids, then remove ethanol, cold in 50 DEG C of revolvings
Lyophilized dry active peptides.The process is easy to operate, reproducible, and Elution range is wide, can be amplified according to actual conditions
Condition realizes industrialized production.
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 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 all be included in the protection scope of the present invention.
Claims (7)
1. suppress the preparation method of the polypeptide extract of cancer cell, it is characterised in that：It the described method comprises the following steps：
1) pretreatment of Optimization of Low Value Fish：By Optimization of Low Value Fish in saline solution boiling, it is cooled to room temperature；
2) preparation of albumen：Optimization of Low Value Fish is put into pressure cooker, in absolute pressure 0.15-0.2MPa boilings, obtains protein liquid；
3) decomposition of albumen：By protein liquid obtained by step 2) in absolute pressure 0.1-0.15MPa boilings, protein breakdown liquid is obtained；
4) isolate and purify：Protein breakdown liquid is filtered through ultrafiltration membrane, trapped fluid is collected, trapped fluid is added according to volume ratio
Into the chromatographic column equipped with 8-10 times of D101 macroreticular resin, impurity is removed with 3-5 times of column volume water elution, then with 5-8 times of column
The ethanol water of volume is eluted, and revolving removes ethanol, dry polypeptide.
2. the preparation method of the polypeptide extract according to claim 1 for suppressing cancer cell, it is characterised in that：The step
1) mass fraction of salt is 22%-25% in, and the mass ratio of Optimization of Low Value Fish and saline solution is 1/1.2-1.5.
3. the preparation method of the polypeptide extract according to claim 1 for suppressing cancer cell, it is characterised in that：The step
1) boiling temperature is 81-87 DEG C in, time 17-21min.
4. the preparation method of the polypeptide extract according to claim 1 for suppressing cancer cell, it is characterised in that：The step
2) boiling temperature is 121-126 DEG C in, digestion time 0.6-1h.
5. the preparation method of the polypeptide extract according to claim 1 for suppressing cancer cell, it is characterised in that：The step
3) boiling temperature is 116-122 DEG C in, digestion time 3.1-3.8h.
6. the preparation method of the polypeptide extract according to claim 1 for suppressing cancer cell, it is characterised in that：The step
4) molecular cut off of ultrafiltration membrane is 1kDa in.
7. the preparation method of the polypeptide extract according to claim 1 for suppressing cancer cell, it is characterised in that：The ethanol
The mass fraction of ethanol is 58%-61% in aqueous solution.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711092034.8A CN107987121A (en) | 2017-11-08 | 2017-11-08 | Suppress the preparation method of the polypeptide extract of cancer cell |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711092034.8A CN107987121A (en) | 2017-11-08 | 2017-11-08 | Suppress the preparation method of the polypeptide extract of cancer cell |
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| CN107987121A true CN107987121A (en) | 2018-05-04 |
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| CN201711092034.8A Withdrawn CN107987121A (en) | 2017-11-08 | 2017-11-08 | Suppress the preparation method of the polypeptide extract of cancer cell |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1628540A (en) * | 2003-12-16 | 2005-06-22 | 威海市宇王水产有限公司 | Production method of fish scale collagen |
| CN104710511A (en) * | 2015-03-18 | 2015-06-17 | 浙江海洋学院 | Iron chelating peptide derived from hairtail fish protein and preparation method and application thereof |
| CN107164447A (en) * | 2017-06-16 | 2017-09-15 | 中国海洋大学 | A kind of method that utilization cod processing accessory substance prepares anti-oxidation peptide |
| CN107279452A (en) * | 2017-07-11 | 2017-10-24 | 浙江丰宇海洋生物制品有限公司 | A kind of processing method of high-quality anchovy protein powder product |
-
2017
- 2017-11-08 CN CN201711092034.8A patent/CN107987121A/en not_active Withdrawn
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1628540A (en) * | 2003-12-16 | 2005-06-22 | 威海市宇王水产有限公司 | Production method of fish scale collagen |
| CN104710511A (en) * | 2015-03-18 | 2015-06-17 | 浙江海洋学院 | Iron chelating peptide derived from hairtail fish protein and preparation method and application thereof |
| CN107164447A (en) * | 2017-06-16 | 2017-09-15 | 中国海洋大学 | A kind of method that utilization cod processing accessory substance prepares anti-oxidation peptide |
| CN107279452A (en) * | 2017-07-11 | 2017-10-24 | 浙江丰宇海洋生物制品有限公司 | A kind of processing method of high-quality anchovy protein powder product |
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Application publication date: 20180504 |
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