CN108148881A - The preparation method of hemoglobin polypeptide powder - Google Patents
The preparation method of hemoglobin polypeptide powder Download PDFInfo
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- CN108148881A CN108148881A CN201711486717.1A CN201711486717A CN108148881A CN 108148881 A CN108148881 A CN 108148881A CN 201711486717 A CN201711486717 A CN 201711486717A CN 108148881 A CN108148881 A CN 108148881A
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- 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
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/795—Porphyrin- or corrin-ring-containing peptides
- C07K14/805—Haemoglobins; Myoglobins
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Abstract
The invention discloses the preparation method of hemoglobin polypeptide powder, the preparation including hemoglobin, the preparation of enzymolysis liquid and the preparation of polypeptide powder.It has the beneficial effect that：The preparation method of polypeptide powder that the present invention is announced is simple, and the polypeptide molecular weight of gained is small, and purity is high, conducive to absorption of human body, can be used directly, and has significant economic benefit.
Description
Technical field
The invention belongs to bioengineering field, more particularly, to the preparation method of hemoglobin polypeptide powder.
Background technology
Hematoglobin protein is with high biological value, 8 kinds of essential amino acids containing needed by human body, Compositional balance,
For high-quality protein source.But current animal blood, particularly sheep blood are most of to be discharged or is abandoned in the form of sewage, is caused big
The valuable nutritional resource of amount is lost in, and cause serious pollution to environment.In sheep blood, hemoglobin accounts for its gross protein and contains
The 60-70% of amount.Bioactive Peptides Derived from Hemoglobin has many particularly important functional activities, as opioid activity, ACE inhibit
Activity, enhancing acllideic i peptide activity, stimulation bacterial growth activity, analgesic activities, antibacterial activity etc..Process hemoglobin not
But contribute to the protection of environment, and a large amount of valuable protein in blood can be made full use of, develop the work(for high added value
It can product.
Invention content
The object of the present invention is to provide the preparation methods of hemoglobin polypeptide powder, and preparation method is simple, the polypeptide of gained
Molecular weight is small, and purity is high, conducive to absorption of human body, can be used directly, and has significant economic benefit.
Technical solution provided by the invention in order to achieve the above object is：The preparation method of hemoglobin polypeptide powder, system
Preparation Method includes the following steps：
The preparation of hemoglobin：Trisodium citrate is added in into sheep blood, the mass ratio of wherein trisodium citrate and sheep blood is 3-5/
100, then stirring centrifuges 6-10min in relative centrifugal force 4000-6000g, removes layer liquid and obtain hemoglobin；
The preparation of enzymolysis liquid：35-45 parts of water are added in into 2-5 parts of hemoglobins, adjust pH to 7-9, are heated to 90-95 DEG C of holding
30-40min is then cooled to 55-60 DEG C, adds in the thiol protease reaction 2-6h for accounting for hemoglobin mass 3%-5%, then
The papain for accounting for hemoglobin mass 2%-5% is added, at 45-55 DEG C, pH hydrolyzes 7-10h under conditions of being 8, reacts
After enzyme deactivation live, filter to take supernatant, obtain enzymolysis liquid, under this condition, using thiol protease and two hatching egg of papain
White enzyme stepwise discretization, obtained polypeptide molecular weight is small, compared with protein, is more easy to digest and assimilate, while sensitization is low, expands
Its application in medicine food；Its solubility in isoelectric point is improved, it is steady under high ionic strength to increase it
It is qualitative, the protein fortification available for food；Enzymolysis also reduces the viscosity of product, changes its rheological equationm of state, facilitates pair
It is processed manufacture；And no harmful substance is mixed into the process, is not had any harmful effect to the health of human body, is belonged to strong
Health green production process；
The preparation of polypeptide powder：By enzymolysis liquid with the flow velocity of 9-10 times of column volume/h by cation exchange resin, then with 9-10
The flow velocity of times column volume/h by anion exchange resin, added in into gained eluent account for the β of supernatant quality 7%-12%-
Cyclic annular monkey essence, it is spray-dried up to polypeptide powder, under this condition, prepared polypeptide has good anti-inflammatory, anti-aging, enhancing
Immunity of organisms, the multiple efficacies such as pre- preventing tumor, beauty, eye-candy have extraordinary suppression to colorectal cancer, lung cancer, breast cancer etc.
System activity, and molecular weight is small, purity is high, conducive to absorption of human body, can be used directly, and has significant economic benefit.
Compared with prior art, beneficial effects of the present invention are：Using thiol protease and two kinds of albumen of papain
Enzyme stepwise discretization, obtained polypeptide molecular weight is small, compared with protein, is more easy to digest and assimilate, while sensitization is low, expand
Its application in medicine food；Its solubility in isoelectric point is improved, increases its stabilization under high ionic strength
Property, the protein fortification available for food；Enzymolysis also reduces the viscosity of product, changes its rheological equationm of state, facilitates to it
It is processed manufacture；And no harmful substance is mixed into the process, to the health of human body does not have any harmful effect, belongs to health
Green production process；Prepared polypeptide has good anti-inflammatory, anti-aging, an enhancing immunity of organisms, pre- preventing tumor, beauty,
The multiple efficacies such as eye-candy have extraordinary inhibitory activity, and molecular weight is small to colorectal cancer, lung cancer, breast cancer etc., and purity is high,
It conducive to absorption of human body, can be used directly, there is significant economic benefit.
Specific embodiment
It is described in further detail by the following examples：
Embodiment 1：The preparation method of hemoglobin polypeptide powder, preparation method include the following steps：
1) preparation of hemoglobin：Trisodium citrate is added in into sheep blood, the mass ratio of wherein trisodium citrate and sheep blood is 3/
100, then stirring centrifuges 6-10min in relative centrifugal force 4000-6000g, removes layer liquid and obtain hemoglobin；
2) preparation of enzymolysis liquid：35 parts of water are added in into 2 parts of hemoglobins, adjust pH to 7, are heated to 90 DEG C of holding 30min, then
55 DEG C are cooled to, the thiol protease reaction 2h for accounting for hemoglobin mass 3% is added in, then adds and account for hemoglobin mass
2% papain, at 45 DEG C, pH hydrolyzes 7h under conditions of being 8, and enzyme deactivation after reaction is lived, and is filtered to take supernatant, is obtained enzyme
Liquid is solved, under this condition, using thiol protease and two kinds of protease stepwise discretizations of papain, obtained polypeptide molecular weight
It is small, it compared with protein, is more easy to digest and assimilate, while sensitization is low, expands its application in medicine food；Improve it
Solubility in isoelectric point increases its stability under high ionic strength, the protein fortification available for food；Enzymolysis
The viscosity of product is also reduced, changes its rheological equationm of state, facilitates and manufacture is processed to it；And in the process without nuisance
Matter is mixed into, and is not had any harmful effect to the health of human body, is belonged to healthy green production process；
3) preparation of polypeptide powder：By enzymolysis liquid with the flow velocity of 9 times of column volume/h by cation exchange resin, then with 9 times of columns
The flow velocity of volume/h by anion exchange resin, added in into gained eluent account for supernatant quality 7% β-ring-type monkey it is smart,
Spray-dried polypeptide powder to obtain the final product, under this condition, prepared polypeptide has good anti-inflammatory, anti-aging, enhancing immunity of organism
Power, the multiple efficacies such as pre- preventing tumor, beauty, eye-candy have colorectal cancer, lung cancer, breast cancer etc. extraordinary inhibitory activity, and
Molecular weight is small, and purity is high, conducive to absorption of human body, can be used directly, and has significant economic benefit.
Embodiment 2：The preparation method of hemoglobin polypeptide powder, preparation method include the following steps：
1) preparation of hemoglobin：Trisodium citrate is added in into sheep blood, the mass ratio of wherein trisodium citrate and sheep blood is 5/
100, then stirring centrifuges 6-10min in relative centrifugal force 4000-6000g, removes layer liquid and obtain hemoglobin；
2) preparation of enzymolysis liquid：45 parts of water are added in into 5 parts of hemoglobins, adjust pH to 9, are heated to 95 DEG C of holding 40min, then
60 DEG C are cooled to, the thiol protease reaction 6h for accounting for hemoglobin mass 5% is added in, then adds and account for hemoglobin mass
5% papain, at 55 DEG C, pH hydrolyzes 10h under conditions of being 8, and enzyme deactivation after reaction is lived, and is filtered to take supernatant, is obtained
Enzymolysis liquid, under this condition, using thiol protease and two kinds of protease stepwise discretizations of papain, obtained peptide molecule
It measures small, compared with protein, is more easy to digest and assimilate, while sensitization is low, expands its application in medicine food；It improves
Its solubility in isoelectric point increases its stability under high ionic strength, the protein fortification available for food；Enzyme
Solution also reduces the viscosity of product, changes its rheological equationm of state, facilitates and manufacture is processed to it；And in the process without harmful
Substance is mixed into, and is not had any harmful effect to the health of human body, is belonged to healthy green production process；
3) preparation of polypeptide powder：By enzymolysis liquid with the flow velocity of 10 times of column volume/h by cation exchange resin, then with 10 times
The flow velocity of column volume/h adds in the β-ring-type monkey for accounting for supernatant quality 12% into gained eluent by anion exchange resin
Essence, it is spray-dried up to polypeptide powder, under this condition, there is prepared polypeptide good anti-inflammatory, anti-aging, enhancing body to exempt from
Epidemic disease power, the multiple efficacies such as pre- preventing tumor, beauty, eye-candy have extraordinary inhibitory activity to colorectal cancer, lung cancer, breast cancer etc.,
And molecular weight is small, purity is high, conducive to absorption of human body, can be used directly, and has significant economic benefit.
Embodiment 3：The preparation method of hemoglobin polypeptide powder, preparation method include the following steps：
1) preparation of hemoglobin：Trisodium citrate is added in into sheep blood, the mass ratio of wherein trisodium citrate and sheep blood is 4/
100, then stirring centrifuges 6-10min in relative centrifugal force 4000-6000g, removes layer liquid and obtain hemoglobin；
2) preparation of enzymolysis liquid：37 parts of water are added in into 3 parts of hemoglobins, adjust pH to 8, are heated to 93 DEG C of holding 34min, then
It is cooled to 57 DEG C, adds in the thiol protease for accounting for hemoglobin mass 5%, add that account for 1 ‰ Propiram of hemoglobin mass more
Sugar and hemoglobin mass 0.3 ‰ 2,4- hexadienoic acids, react 3h, are subsequently added into the wood for accounting for hemoglobin mass 2%
Melon protease, at 48 DEG C, pH hydrolyzes 8h under conditions of being 8, and enzyme deactivation after reaction is lived, and is filtered to take supernatant, is obtained enzymolysis liquid, institute
2,4- hexadienoic acids, the pulullan polysaccharide added in has synergistic effect, enhances cysteine contained by thiol protease
Activity improves the ability of its catalyzing hydrolysis peptide bond, reduces the molecular weight of the polypeptide of enzymolysis gained, improves its economy effect
Benefit；
3) preparation of polypeptide powder：By enzymolysis liquid with the flow velocity of 9 times of column volume/h by cation exchange resin, then with 10 times of columns
The flow velocity of volume/h adds in the β-ring-type monkey for accounting for supernatant quality 12% into gained eluent by anion exchange resin
Essence, it is spray-dried up to polypeptide powder, under this condition, there is prepared polypeptide good anti-inflammatory, anti-aging, enhancing body to exempt from
Epidemic disease power, the multiple efficacies such as pre- preventing tumor, beauty, eye-candy have extraordinary inhibitory activity to colorectal cancer, lung cancer, breast cancer etc.,
And molecular weight is small, purity is high, conducive to absorption of human body, can be used directly, and has significant economic benefit.
Embodiment of above is merely to illustrate the present invention, and not limitation of the present invention, the ordinary skill people of this field
Member without departing from the spirit and scope of the present invention, can also make a variety of changes and modification.Therefore, it is all equivalent
Technical solution also belong to scope of the invention, scope of patent protection of the invention should be defined by the claims.
Claims (6)
1. the preparation method of hemoglobin polypeptide powder, the preparation including hemoglobin, the preparation of enzymolysis liquid and the system of polypeptide powder
It is standby, it is characterised in that：The preparation method of the enzymolysis liquid is：35-45 parts of water are added in into hemoglobin, adjust pH to 7-9, heating
To 90-95 DEG C of holding 30-40min, 55-60 DEG C is then cooled to, thiol protease reaction is added in, then adds Papain
Enzyme, at 45-55 DEG C, pH reacts under conditions of being 8, and enzyme deactivation after reaction is lived, and is filtered to take supernatant, is obtained enzymolysis liquid.
2. the preparation method of hemoglobin polypeptide powder according to claim 1, it is characterised in that：The system of the hemoglobin
Preparation Method is：Trisodium citrate is added in into sheep blood, is stirred, then 6-10min is centrifuged in relative centrifugal force 4000-6000g, takes
Lower floor's liquid obtains hemoglobin.
3. the preparation method of hemoglobin polypeptide powder according to claim 1, it is characterised in that：The preparation of the polypeptide powder
Method is：By enzymolysis liquid with the flow velocity of 9-10 times of column volume/h by cation exchange resin, then with 9-10 times of column volume/h
Flow velocity by anion exchange resin, added in into gained eluent account for supernatant quality 7%-12% β-ring-type monkey it is smart,
Spray-dried polypeptide powder to obtain the final product.
4. the preparation method of hemoglobin polypeptide powder according to claim 1, it is characterised in that：The thiol protease
Addition accounts for the 3%-5% of hemoglobin mass, and the thiol protease reaction time is 2-6h.
5. the preparation method of hemoglobin polypeptide powder according to claim 1, it is characterised in that：The papain
Addition accounts for the 2%-5% of hemoglobin mass, and the papain reaction time is 7-10h.
6. the preparation method of hemoglobin polypeptide powder according to claim 2, it is characterised in that：The trisodium citrate with
The mass ratio of sheep blood is 3-5/100.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201711486717.1A CN108148881A (en) | 2017-12-29 | 2017-12-29 | The preparation method of hemoglobin polypeptide powder |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201711486717.1A CN108148881A (en) | 2017-12-29 | 2017-12-29 | The preparation method of hemoglobin polypeptide powder |
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| CN201711486717.1A Withdrawn CN108148881A (en) | 2017-12-29 | 2017-12-29 | The preparation method of hemoglobin polypeptide powder |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109793885A (en) * | 2019-03-13 | 2019-05-24 | 湖北瑞邦生物科技有限公司 | It is a kind of for preventing or alleviating the preparation method of the complex polypeptide of anaemia |
Citations (6)
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|---|---|---|---|---|
| WO2002091860A1 (en) * | 2001-05-14 | 2002-11-21 | Vasily Petrovich Andreichuk | Biologically active food additive and biologically active forage additive for preventing iodine deficiency and optimising iodine metabolism, food and forage products containing said additives |
| CN1454494A (en) * | 2003-04-29 | 2003-11-12 | 东北农业大学 | Albumen oligo peptide and preparing method thereof |
| CN101744092A (en) * | 2010-01-21 | 2010-06-23 | 辽源市麒鸣生物技术综合开发有限公司 | Sheep hemoglobin polypeptide powder and preparation method thereof |
| WO2011122740A1 (en) * | 2010-04-01 | 2011-10-06 | Gendocs, Inc. | Environmental stress-inducible 557 promoter isolated from rice and uses thereof |
| US20140220163A1 (en) * | 2014-02-04 | 2014-08-07 | Hamzeh Soleimani Babadi | Powder mixture composition of natural materials for controlling deficiencies in polypeptide alpha and beta hemoglobin chains |
| CN104745663A (en) * | 2015-03-24 | 2015-07-01 | 合肥学院 | Method for comprehensively utilizing porcine hemoglobin |
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2017
- 2017-12-29 CN CN201711486717.1A patent/CN108148881A/en not_active Withdrawn
Patent Citations (6)
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|---|---|---|---|---|
| WO2002091860A1 (en) * | 2001-05-14 | 2002-11-21 | Vasily Petrovich Andreichuk | Biologically active food additive and biologically active forage additive for preventing iodine deficiency and optimising iodine metabolism, food and forage products containing said additives |
| CN1454494A (en) * | 2003-04-29 | 2003-11-12 | 东北农业大学 | Albumen oligo peptide and preparing method thereof |
| CN101744092A (en) * | 2010-01-21 | 2010-06-23 | 辽源市麒鸣生物技术综合开发有限公司 | Sheep hemoglobin polypeptide powder and preparation method thereof |
| WO2011122740A1 (en) * | 2010-04-01 | 2011-10-06 | Gendocs, Inc. | Environmental stress-inducible 557 promoter isolated from rice and uses thereof |
| US20140220163A1 (en) * | 2014-02-04 | 2014-08-07 | Hamzeh Soleimani Babadi | Powder mixture composition of natural materials for controlling deficiencies in polypeptide alpha and beta hemoglobin chains |
| CN104745663A (en) * | 2015-03-24 | 2015-07-01 | 合肥学院 | Method for comprehensively utilizing porcine hemoglobin |
Non-Patent Citations (3)
| Title |
|---|
| 李良铸等: "《最新生化药物制备技术》", 30 June 2016 * |
| 郝利平等: "《食品添加剂》", 31 August 2002, 北京：中国农业大学出版社 * |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109793885A (en) * | 2019-03-13 | 2019-05-24 | 湖北瑞邦生物科技有限公司 | It is a kind of for preventing or alleviating the preparation method of the complex polypeptide of anaemia |
| CN109793885B (en) * | 2019-03-13 | 2022-07-08 | 湖北瑞邦生物科技有限公司 | Preparation method of composite polypeptide for preventing or relieving anemia |
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Application publication date: 20180612 |
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