CN105132399A - Method for extracting plasma protein powder from pig blood - Google Patents
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- CN105132399A CN105132399A CN201510628766.9A CN201510628766A CN105132399A CN 105132399 A CN105132399 A CN 105132399A CN 201510628766 A CN201510628766 A CN 201510628766A CN 105132399 A CN105132399 A CN 105132399A
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- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
- C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
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- C07K14/4717—Plasma globulins, lactoglobulin
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Abstract
The invention provides a method for extracting plasma protein powder from pig blood. The method comprises the following steps of preparing anticoagulation pig blood; separating plasma from haemocyte; depositing the plasma to collect prothrombin; preparing crude thrombin liquid; preparing fine thrombin products; breaking walls and performing hemolysis of the haemocyte; performing enzymolysis of hyperglobulinemia by segment; preparing hydrolyzed protein of the pig blood. The method has the beneficial effects that the amino acid content of the hydrolyzed protein of the pig blood is more than 90% of protein, the hydrolyzed protein of the pig blood is easy to absorb and utilize, raw materials are fully utilized, and two high added-value products can be produced by once charging. In the whole process of production, organic solvents with large toxicity are not used, the defects of product singleness, solvent residue and the like are overcome, and the production cost is reduced.
Description
Technical field
The invention belongs to the separation and extraction technology field of biologically active substance, particularly relate to the method that a boar blood extracts plasma protein powder.
Background technology
Pig blood is a kind of high-quality protein, and nutritionist is called liquid meat.Containing the most nutritive ingredient that needed by human body is wanted in pig blood: the indispensable trace element of human body such as protein, amino acid, VITAMIN, sugar and sodium, potassium, iron, calcium.Wherein protein 18.9%, non-protein organism 1.2%, mineral substance 0.9%, moisture 79%, in addition, its amino acid Compositional balance, containing the total amount of 8 kinds of indispensable amino acids of needed by human body higher than human milk and shell egg, particularly the content of Methionin is up to 9%, meets Food and Argriculture OrganizationFAO, pattern is recommended by the World Health Organization (FAO/WHO) expert group.
Shi Zhu blood resource big country of China, but on pig blood resource develops, lag behind many developed countries, due to the not easily digested absorption and color is comparatively dark, the heavier palatability of the blood smell is poor of the porcine haemoglobin through simple processing, so for a long time, much pig blood pours trench into as waste, both huge waste was caused, again contaminate environment.
Start from the beginning of last century to the exploitation of animal blood abroad, the initial stage is mainly used in fodder industry.Along with the development of new and high technology, domestic and international many research departments especially emphasis biochemical technology processing pig blood are used for the research of the aspect such as food, pharmacy.Current blood products reaches a certain scale in health products trade, and industrialization prospect is good.In recent years, China's pig blood deep development also achieves impressive progress, and pig blood, as a nutrition treasure-house, has obtained studying extremely widely, not lower tens kinds of the producing process flow process proposed.From direct edible pork blood bean curd, to enzymolysis Swine blood meal, the production of compound amino acid powder, has achieved considerable progress, but still can not meet the needs of the mankind.Problems existing is most of technique also imperfection, if any the large length consuming time of process energy consumption, the technique that has can only produce single product, the technique yield technique that is low, that have that has uses a large amount of organic solvent, there is environmental protection defect etc.And the present invention overcomes above-mentioned shortcoming, the pig blood that utilizes having invented zero release produces zymoplasm and the protelytic green method of pig watery blood simultaneously.
Summary of the invention
The object of this invention is to provide a boar blood and extract the method for plasma protein powder, solve above-mentioned one or multiple of the prior art.
The invention provides the method that a boar blood extracts plasma protein powder, comprise the steps:
1) anti-freezing pig blood is prepared: get the qualified fresh pig blood of quarantine and put into the first container, add in advance in this container the Trisodium Citrate of 1.0%, 1.0% disodium ethylene diamine tetraacetate, 1.0% EDTA-2K, 0.6% Sodium hexametaphosphate 99 and 0.8% trisodium phosphate composition antithrombotics, stir, at 7 ~ 8 DEG C, canning 42h;
2) blood plasma, blood cell are separated: at 7 ~ 8 DEG C, after canning 42h, anti-freezing pig blood upper strata is colourless transparent liquid is blood plasma, and lower floor's scarlet thick liquid is blood cell, is drawn onto in second container with siphonage by upper plasma; Remaining blood cell through rotating speed be 2500r/min centrifugal after, upper plasma is merged into cryogenic freezing in second container for subsequent use, blood cell then spends the night in chilled storage to the 3rd container stand-by;
3) blood plasma precipitates collects thrombogen: after the blood plasma unfreezing in second container, after the White Flocculus that filtering upper strata is floating, use 0.9% normal saline dilution, stand at low temperature is spent the night, and namely collecting precipitation obtains thrombogen;
4) blood coagulation crude enzyme liquid preparation: the thrombogen of collection being precipitated and dissolved in pH value is in the phosphoric acid buffer of 7.0, uses 1.0%CaCl
2solution activates thrombogen, and namely collecting by filtration filtrate obtain zymoplasm crude enzyme liquid;
5) zymoplasm fine work preparation: be the nanofiltration membrane treatment of 1.2nm by zymoplasm crude enzyme liquid aperture, after concentrated, the 1/25-1/30 of original volume collects trapped fluid, further by trapped fluid spraying dry, obtains plasma protein powder;
6) blood cell broken wall, haemolysis: the appropriate physiological saline adding corpuscle volume in the 3rd container, makes protein content in liquid be 20%, be warming up to 60 DEG C, carry out ultrasonication 12min;
7) hyperglobulinemia subsection enzymolysis: by 4.0mol/LNaOH adjust ph to 8.0, temperature is risen to 40-45 DEG C, add appropriate serrapeptass, be incubated after 2 hours, then be warming up to 50 DEG C, be incubated 3 hours; Adjust ph to 9.0, adds appropriate compound protease, is incubated and hyperglobulinemia is fully hydrolyzed in 4 hours;
8) pig blood protolysate preparation: after enzymolysis terminates, be cooled to normal temperature, namely can be made into liquid pig blood protolysate with the membrane filtration of 0.20um is degerming, make globulin powder by after this liquid spray drying further.
The invention has the beneficial effects as follows: in the pig blood protolysate that the method obtains, aminoacids content accounts for more than 90% of protein, is easy to absorb, and takes full advantage of raw material, once feeds intake and can produce two kinds of high value added products.Whole production process avoids the organic solvent using toxicity large, overcomes the defects such as product is single, dissolvent residual, reduces production cost.
In some embodiments, in step 6) in, the condition of ultrasonication is ultrasonic power 250W.
In some embodiments, in step 7) in, described compound protease is mixed by a certain percentage by trypsinase, Sumizyme MP, papoid, bromeline.
Embodiment
The invention provides the method that a boar blood extracts plasma protein powder, below in conjunction with embodiment, the present invention described in detail:
Embodiment 1
In the present embodiment, compound protease is mixed by a certain percentage by trypsinase, Sumizyme MP, papoid, bromeline.Concrete steps are:
1) anti-freezing pig blood is prepared: get the qualified fresh pig blood of quarantine and put into the first container, add in advance in this container the Trisodium Citrate of 1.0%, 1.0% disodium ethylene diamine tetraacetate, 1.0% EDTA-2K, 0.6% Sodium hexametaphosphate 99 and 0.8% trisodium phosphate composition antithrombotics, stir, at 7 DEG C, canning 42h;
2) blood plasma, blood cell are separated: at 7 DEG C, after canning 42h, anti-freezing pig blood upper strata is colourless transparent liquid is blood plasma, and lower floor's scarlet thick liquid is blood cell, is drawn onto in second container with siphonage by upper plasma; Remaining blood cell through rotating speed be 2500r/min centrifugal after, upper plasma is merged into cryogenic freezing in second container for subsequent use, blood cell then spends the night in chilled storage to the 3rd container stand-by;
3) blood plasma precipitates collects thrombogen: after the blood plasma unfreezing in second container, after the White Flocculus that filtering upper strata is floating, use 0.9% normal saline dilution, stand at low temperature is spent the night, and namely collecting precipitation obtains thrombogen; 4) blood coagulation crude enzyme liquid preparation: the thrombogen of collection being precipitated and dissolved in pH value is in the phosphoric acid buffer of 7.0, uses 1.0%CaCl
2solution activates thrombogen, and namely collecting by filtration filtrate obtain zymoplasm crude enzyme liquid;
5) zymoplasm fine work preparation: be the nanofiltration membrane treatment of 1.2nm by zymoplasm crude enzyme liquid aperture, after concentrated, 1/30 of original volume collects trapped fluid, further by trapped fluid spraying dry, obtains plasma protein powder;
6) blood cell broken wall, haemolysis: the appropriate physiological saline adding corpuscle volume in the 3rd container, makes protein content in liquid be 20%, be warming up to 60 DEG C, carries out the ultrasonication 12min that power is 250W;
7) hyperglobulinemia subsection enzymolysis: by 4.0mol/LNaOH adjust ph to 8.0, temperature is risen to 40 DEG C, add appropriate serrapeptass, be incubated after 2 hours, then be warming up to 50 DEG C, be incubated 3 hours; Adjust ph to 9.0, adds appropriate compound protease, is incubated and hyperglobulinemia is fully hydrolyzed in 4 hours;
8) pig blood protolysate preparation: after enzymolysis terminates, be cooled to normal temperature, namely can be made into liquid pig blood protolysate with the membrane filtration of 0.20um is degerming, make globulin powder by after this liquid spray drying further.
Embodiment 2
In the present embodiment, compound protease is mixed by a certain percentage by trypsinase, Sumizyme MP, papoid, bromeline.Concrete steps are:
1) anti-freezing pig blood is prepared: get the qualified fresh pig blood of quarantine and put into the first container, add in advance in this container the Trisodium Citrate of 1.0%, 1.0% disodium ethylene diamine tetraacetate, 1.0% EDTA-2K, 0.6% Sodium hexametaphosphate 99 and 0.8% trisodium phosphate composition antithrombotics, stir, at 7 DEG C, canning 42h;
2) blood plasma, blood cell are separated: at 7 DEG C, after canning 42h, anti-freezing pig blood upper strata is colourless transparent liquid is blood plasma, and lower floor's scarlet thick liquid is blood cell, is drawn onto in second container with siphonage by upper plasma; Remaining blood cell through rotating speed be 2500r/min centrifugal after, upper plasma is merged into cryogenic freezing in second container for subsequent use, blood cell then spends the night in chilled storage to the 3rd container stand-by;
3) blood plasma precipitates collects thrombogen: after the blood plasma unfreezing in second container, after the White Flocculus that filtering upper strata is floating, use 0.9% normal saline dilution, stand at low temperature is spent the night, and namely collecting precipitation obtains thrombogen;
4) blood coagulation crude enzyme liquid preparation: the thrombogen of collection being precipitated and dissolved in pH value is in the phosphoric acid buffer of 7.0, uses 1.0%CaCl
2solution activates thrombogen, and namely collecting by filtration filtrate obtain zymoplasm crude enzyme liquid;
5) zymoplasm fine work preparation: be the nanofiltration membrane treatment of 1.2nm by zymoplasm crude enzyme liquid aperture, after concentrated, 1/27 of original volume collects trapped fluid, further by trapped fluid spraying dry, obtains plasma protein powder;
6) blood cell broken wall, haemolysis: the appropriate physiological saline adding corpuscle volume in the 3rd container, makes protein content in liquid be 20%, be warming up to 60 DEG C, carries out the ultrasonication 12min that power is 250W;
7) hyperglobulinemia subsection enzymolysis: by 4.0mol/LNaOH adjust ph to 8.0, temperature is risen to 42 DEG C, add appropriate serrapeptass, be incubated after 2 hours, then be warming up to 50 DEG C, be incubated 3 hours; Adjust ph to 9.0, adds appropriate compound protease, is incubated and hyperglobulinemia is fully hydrolyzed in 4 hours;
8) pig blood protolysate preparation: after enzymolysis terminates, be cooled to normal temperature, namely can be made into liquid pig blood protolysate with the membrane filtration of 0.20um is degerming, make globulin powder by after this liquid spray drying further.
Embodiment 3
In the present embodiment, compound protease is mixed by a certain percentage by trypsinase, Sumizyme MP, papoid, bromeline.Concrete steps are:
1) anti-freezing pig blood is prepared: get the qualified fresh pig blood of quarantine and put into the first container, add in advance in this container the Trisodium Citrate of 1.0%, 1.0% disodium ethylene diamine tetraacetate, 1.0% EDTA-2K, 0.6% Sodium hexametaphosphate 99 and 0.8% trisodium phosphate composition antithrombotics, stir, at 8 DEG C, canning 42h;
2) blood plasma, blood cell are separated: at 8 DEG C, after canning 42h, anti-freezing pig blood upper strata is colourless transparent liquid is blood plasma, and lower floor's scarlet thick liquid is blood cell, is drawn onto in second container with siphonage by upper plasma; Remaining blood cell through rotating speed be 2500r/min centrifugal after, upper plasma is merged into cryogenic freezing in second container for subsequent use, blood cell then spends the night in chilled storage to the 3rd container stand-by;
3) blood plasma precipitates collects thrombogen: after the blood plasma unfreezing in second container, after the White Flocculus that filtering upper strata is floating, use 0.9% normal saline dilution, stand at low temperature is spent the night, and namely collecting precipitation obtains thrombogen;
4) blood coagulation crude enzyme liquid preparation: the thrombogen of collection being precipitated and dissolved in pH value is in the phosphoric acid buffer of 7.0, uses 1.0%CaCl
2solution activates thrombogen, and namely collecting by filtration filtrate obtain zymoplasm crude enzyme liquid;
5) zymoplasm fine work preparation: be the nanofiltration membrane treatment of 1.2nm by zymoplasm crude enzyme liquid aperture, after concentrated, 1/25 of original volume collects trapped fluid, further by trapped fluid spraying dry, obtains plasma protein powder;
6) blood cell broken wall, haemolysis: the appropriate physiological saline adding corpuscle volume in the 3rd container, makes protein content in liquid be 20%, be warming up to 60 DEG C, carries out the ultrasonication 12min that power is 250W;
7) hyperglobulinemia subsection enzymolysis: by 4.0mol/LNaOH adjust ph to 8.0, temperature is risen to 45 DEG C, add appropriate serrapeptass, be incubated after 2 hours, then be warming up to 50 DEG C, be incubated 3 hours; Regulate pH to 9.0, add appropriate compound protease, be incubated and hyperglobulinemia be fully hydrolyzed in 4 hours;
8) pig blood protolysate preparation: after enzymolysis terminates, be cooled to normal temperature, namely can be made into liquid pig blood protolysate with the membrane filtration of 0.20um is degerming, make globulin powder by after this liquid spray drying further.
Above-described is only some embodiments of the present invention.For the person of ordinary skill of the art, without departing from the concept of the premise of the invention, can also make some distortion and improvement, these all belong to protection scope of the present invention.
Claims (3)
1. a boar blood extracts the method for plasma protein powder, wherein, comprises the steps:
1) anti-freezing pig blood is prepared: get the qualified fresh pig blood of quarantine and put into the first container, add in advance in this container the Trisodium Citrate of 1.0%, 1.0% disodium ethylene diamine tetraacetate, 1.0% EDTA-2K, 0.6% Sodium hexametaphosphate 99 and 0.8% trisodium phosphate composition antithrombotics, stir, at 7 ~ 8 DEG C, canning 42h;
2) blood plasma, blood cell are separated: at 7 ~ 8 DEG C, after canning 42h, anti-freezing pig blood upper strata is colourless transparent liquid is blood plasma, and lower floor's scarlet thick liquid is blood cell, is drawn onto in second container with siphonage by upper plasma; Remaining blood cell through rotating speed be 2500r/min centrifugal after, upper plasma is merged into cryogenic freezing in second container for subsequent use, blood cell then spends the night in chilled storage to the 3rd container stand-by;
3) blood plasma precipitates collects thrombogen: after the blood plasma unfreezing in second container, after the White Flocculus that filtering upper strata is floating, use 0.9% normal saline dilution, stand at low temperature is spent the night, and namely collecting precipitation obtains thrombogen;
4) blood coagulation crude enzyme liquid preparation: the thrombogen of collection being precipitated and dissolved in pH value is in the phosphoric acid buffer of 7.0, uses 1.0%CaCl
2solution activates thrombogen, and namely collecting by filtration filtrate obtain zymoplasm crude enzyme liquid;
5) zymoplasm fine work preparation: be the nanofiltration membrane treatment of 1.2nm by zymoplasm crude enzyme liquid aperture, after concentrated, the 1/25-1/30 of original volume collects trapped fluid, further by trapped fluid spraying dry, obtains plasma protein powder;
6) blood cell broken wall, haemolysis: the appropriate physiological saline adding corpuscle volume in the 3rd container, makes protein content in liquid be 20%, be warming up to 60 DEG C, carry out ultrasonication 12min;
7) hyperglobulinemia subsection enzymolysis: by 4.0mol/LNaOH adjust ph to 8.0, temperature is risen to 40-45 DEG C, add appropriate serrapeptass, be incubated after 2 hours, then be warming up to 50 DEG C, be incubated 3 hours; Adjust ph to 9.0, adds appropriate compound protease, is incubated and hyperglobulinemia is fully hydrolyzed in 4 hours;
8) pig blood protolysate preparation: after enzymolysis terminates, be cooled to normal temperature, namely can be made into liquid pig blood protolysate with the membrane filtration of 0.20um is degerming, make globulin powder by after this liquid spray drying further.
2. a boar blood according to claim 1 extracts the method for plasma protein powder, wherein, in step 6) in, the condition of ultrasonication is ultrasonic power 250W.
3. a boar blood according to claim 1 extracts the method for plasma protein powder, wherein, in step 7) in, described compound protease is mixed by a certain percentage by trypsinase, Sumizyme MP, papoid, bromeline.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105950576A (en) * | 2016-05-26 | 2016-09-21 | 成都远睿生物技术有限公司 | Method for extracting multiple proteins from bovine blood |
| CN106191188A (en) * | 2016-08-09 | 2016-12-07 | 广东海洋大学 | A kind of method for hydrolysis of king crab plasma protein |
| CN106497903A (en) * | 2016-09-26 | 2017-03-15 | 河北大安制药有限公司 | A kind of technique for purifying blood coagulation proenzyme complex |
| CN107245422A (en) * | 2017-08-03 | 2017-10-13 | 重庆市药物种植研究所 | A kind of deer-blood wine and preparation method thereof |
| CN107699555A (en) * | 2017-11-17 | 2018-02-16 | 浙江丰安生物制药有限公司 | A kind of preparation method of pig blood fibrin ferment |
| CN107836701A (en) * | 2017-09-04 | 2018-03-27 | 韩山师范学院 | A kind of pig blood anti-coagulants |
| CN110042139A (en) * | 2019-05-05 | 2019-07-23 | 王福芳 | A kind of preparation method and applications of animal blood activity complex peptides |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105950576A (en) * | 2016-05-26 | 2016-09-21 | 成都远睿生物技术有限公司 | Method for extracting multiple proteins from bovine blood |
| CN106191188A (en) * | 2016-08-09 | 2016-12-07 | 广东海洋大学 | A kind of method for hydrolysis of king crab plasma protein |
| CN106191188B (en) * | 2016-08-09 | 2020-01-07 | 广东海洋大学 | Hydrolysis method of limulus plasma protein |
| CN106497903A (en) * | 2016-09-26 | 2017-03-15 | 河北大安制药有限公司 | A kind of technique for purifying blood coagulation proenzyme complex |
| CN106497903B (en) * | 2016-09-26 | 2018-07-20 | 河北大安制药有限公司 | A kind of technique for purifying blood coagulation proenzyme compound |
| CN107245422A (en) * | 2017-08-03 | 2017-10-13 | 重庆市药物种植研究所 | A kind of deer-blood wine and preparation method thereof |
| CN107245422B (en) * | 2017-08-03 | 2021-04-13 | 重庆市药物种植研究所 | Deer blood wine and preparation method thereof |
| CN107836701A (en) * | 2017-09-04 | 2018-03-27 | 韩山师范学院 | A kind of pig blood anti-coagulants |
| CN107699555A (en) * | 2017-11-17 | 2018-02-16 | 浙江丰安生物制药有限公司 | A kind of preparation method of pig blood fibrin ferment |
| CN110042139A (en) * | 2019-05-05 | 2019-07-23 | 王福芳 | A kind of preparation method and applications of animal blood activity complex peptides |
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Application publication date: 20151209 |