CN107840869A - Alleviate the preparation method of the active peptides of knee osteoarthritis - Google Patents
Alleviate the preparation method of the active peptides of knee osteoarthritis Download PDFInfo
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- CN107840869A CN107840869A CN201711092051.1A CN201711092051A CN107840869A CN 107840869 A CN107840869 A CN 107840869A CN 201711092051 A CN201711092051 A CN 201711092051A CN 107840869 A CN107840869 A CN 107840869A
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- preparation
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- low value
- active peptides
- knee osteoarthritis
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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
- C07K1/145—Extraction; Separation; Purification by extraction or solubilisation
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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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- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention discloses the preparation method for the active peptides for alleviating knee osteoarthritis, method includes the pretreatment of Optimization of Low Value Fish, the preparation of albumen, the decomposition of albumen and isolated and purified.Have the beneficial effect that：Addition of this method without catalyst or enzyme, avoids separating step；Salt solution boiling Optimization of Low Value Fish, the effect of sterilization is served to Optimization of Low Value Fish, 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 alleviate knee osteoarthritis.
Description
Technical field
The present invention relates to technical field of biological product preparation, more particularly, to the system for the active peptides for alleviating knee osteoarthritis
Preparation Method.
Background technology
Knee osteoarthritis is using cartilage degeneration as the bone and sclerotin of core, synovial membrane, capsular ligament and joint other structures
Multi-faceted, multi-level, different degrees of chronic inflammation.Its clinical manifestation is mostly gonalgia, swelling, dysfunction, pass
Save thick, Varus deformity etc., knee osteoarthritis is mostly the elderly, be the disease of asthenia in origin and asthenia in superficiality, in addition due to knee joint ache
Pain and long-term lacking activity, cause local QI-blood circulation deficiency, and muscle is become homeless foster, numbness of aching, long easy to cause myoatrophy and sclerotin
It is loose.
The content of the invention
It is an object of the invention to provide the preparation method for the active peptides for alleviating knee osteoarthritis, this method is without catalyst
Or the addition of enzyme, separating step, and the high income of gained active peptides are avoided, can effectively alleviate knee osteoarthritis.
The present invention is directed to the problem of being mentioned in background technology, and the technical scheme taken is：Alleviate the activity of knee osteoarthritis
The preparation method of polypeptide, 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, room temperature is cooled to, 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 salt solution boiling Optimization of Low Value Fish, the effect of sterilization is served 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, protein liquid is obtained, 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, 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 milipore filter 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 removes in 50 DEG C of revolvings
Remove ethanol, be freeze-dried to obtain active peptides.The process operation is simple, 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 synergy with ethanol, improves the yield of active peptides during revolving.
Compared with prior art, the advantage of the invention is that：The active peptides disclosed by the invention for alleviating knee osteoarthritis
Preparation method include the pretreatment of Optimization of Low Value Fish, the preparation of albumen, the decomposition of albumen and isolate and purify, this method is without catalyst
Or the addition of enzyme, avoid separating step；Salt solution boiling Optimization of Low Value Fish, the effect of sterilization is served 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.
Embodiment
Below by embodiment, the present invention will be further described：
Embodiment 1：
Alleviate the preparation method of the active peptides of knee osteoarthritis, 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 salt solution boiling Optimization of Low Value Fish, the effect of sterilization, and the albumen in Optimization of Low Value Fish at this temperature are served 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, obtained
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 milipore filter 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 operation is simple, reproducible, and Elution range is wide, can be put according to actual conditions
Big condition realizes industrialized production.
Embodiment 2：
Alleviate the preparation method of the active peptides of knee osteoarthritis, the described method comprises the following steps：It is the pre- of Optimization of Low Value Fish first
Processing：By Optimization of Low Value Fish boiling in the saline solution that mass fraction is 25%, boiling temperature is 87 DEG C, 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,
The boiling 1h at absolute pressure 0.2MPa, 126 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
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 milipore filter 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 operation is simple, reproducible, and Elution range is wide, can be amplified according to actual conditions
Condition realizes industrialized production.
Embodiment 3：
Alleviate the preparation method of the active peptides of knee osteoarthritis, the described method comprises the following steps：It is the pre- of Optimization of Low Value Fish first
Processing：By Optimization of Low Value Fish boiling in the saline solution that mass fraction is 24%, boiling temperature is 86 DEG C, 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,
The boiling 1h at absolute pressure 0.2MPa, 125 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
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 milipore filter 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 operation is simple, 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 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.
Claims (7)
1. alleviate the preparation method of the active peptides of knee osteoarthritis, 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, room temperature is cooled to；
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 milipore filter, 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 post
The ethanol water of volume is eluted, and revolving removes ethanol, dry polypeptide.
2. the preparation method of the active peptides according to claim 1 for alleviating knee osteoarthritis, it is characterised in that：The step
It is rapid 1) in the mass fraction of salt be 22%-25%, the mass ratio of Optimization of Low Value Fish and saline solution is 1/1.2-1.5.
3. the preparation method of the active peptides according to claim 1 for alleviating knee osteoarthritis, it is characterised in that：The step
It is rapid 1) in boiling temperature be 81-87 DEG C, time 17-21min.
4. the preparation method of the active peptides according to claim 1 for alleviating knee osteoarthritis, it is characterised in that：The step
It is rapid 2) in boiling temperature be 121-126 DEG C, digestion time 0.6-1h.
5. the preparation method of the active peptides according to claim 1 for alleviating knee osteoarthritis, it is characterised in that：The step
It is rapid 3) in boiling temperature be 116-122 DEG C, digestion time 3.1-3.8h.
6. the preparation method of the active peptides according to claim 1 for alleviating knee osteoarthritis, it is characterised in that：The step
It is rapid 4) in the molecular cut off of milipore filter be 1kDa.
7. the preparation method of the active peptides according to claim 1 for alleviating knee osteoarthritis, it is characterised in that：The second
The mass fraction of ethanol is 58%-61% in alcohol solution.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201711092051.1A CN107840869A (en) | 2017-11-08 | 2017-11-08 | Alleviate the preparation method of the active peptides of knee osteoarthritis |
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| CN201711092051.1A CN107840869A (en) | 2017-11-08 | 2017-11-08 | Alleviate the preparation method of the active peptides of knee osteoarthritis |
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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 CN201711092051.1A patent/CN107840869A/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: 20180327 |