CN106086140A - A kind of method that microwave-assisted and membrane filtration prepare fresh-water fishes hemepeptide - Google Patents
A kind of method that microwave-assisted and membrane filtration prepare fresh-water fishes hemepeptide Download PDFInfo
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Abstract
The invention belongs to technical field of bioengineering, be specifically related to a kind of method that microwave-assisted prepares fresh-water fishes hemepeptide with membrane filtration.A kind of method that microwave-assisted and membrane filtration prepare fresh-water fishes hemepeptide, comprises the following steps: anticoagulant process, precipitation, enzymolysis, breaking cellular wall, microwave-assisted subsection enzymolysis, enzymolysis 1, enzymolysis 2, enzyme denaturing are lived, filtered and be spray-dried.The microwave-assisted of the present invention and membrane filtration are prepared the method for fresh-water fishes hemepeptide and are had that yield is high, purity is high, solvent consumption is few, low cost and the advantage that is prone to industrialized production.The whole technological process of the present invention is short, the time is short, and energy consumption is low, pollution less, does not use toxic reagent, non-pollutant discharge, reaches to clean productive target.
Description
Technical field
The invention belongs to technical field of bioengineering, be specifically related to a kind of microwave-assisted and prepare fresh-water fishes hemepeptide with membrane filtration
Method.
Background technology
In fresh-water fishes blood, erythrocytic protein content is up to 38%, accounts for more than the 75% of whole blood Tot Prot, and wherein 92%
Being above hemoglobin, therefore fresh-water fishes blood cell is a kind of excellent animal protein source.But owing to blood cell powder containing haemachrome,
Making blood cell powder is dark red, and with the heaviest smell of blood, bad being difficult to accepts sense organ, and its economic value added is the highest
Haemachrome is that the form with ferrous porphyrin is combined formation hemoglobin with four globins, is present in erythrocyte.?
In food industry, the colour former nitrate in the alternative meat products of haemachrome and synthetic food color, use haemachrome to reduce
The carcinogenesis of nitrite.In pharmaceuticals industry, haemachrome can as semi-synthetic bilirubin raw material, and be anticancer specially good effect
Medicine.Haemachrome, clinically as iron supplementary, can treat the anemia caused by iron deficiency, and this haemachrome iron supplementary can be direct
Being absorbed by the body, absorbance is up to 10～20%, has and accumulates poisoning without internal ferrum and the advantages such as untoward reaction such as gastrointestinal irritation.
The bioactive peptide obtained by protein digestion, is one of current most active field of performance animal nutrient research.Fill
Point utilize the abundant protein resource in fresh-water fishes blood, utilize the modern biotechnology zymolysis technique can be by the globin in fresh-water fishes blood
It is converted into the protein fragments of little molecule and there is bioactive little peptide.Little Molecular hemoglobin peptide does not only have and well dissolves
Property, low viscosity, anti-gel formative, and digest and assimilate fast in vivo, protein utilization is high, has low antigenicity, will not
Produce anaphylaxis.Biological enzymolysis technology reaction condition is gentle, easily operation, energy consumption are low, can avoid because of violent the caused battalion of operation
The loss formed point, and improve the content of essential amino acids and delicious amino acid, improve the palatability of protein, improve
Digestibility.
Therefore, become there is bioactive peptide matters by the hemoglobin enzymolysis in fish hemocyte, will greatly meet people
Utilization rate to functional raising protein resource.
The information being disclosed in this background section is merely intended to increase the understanding of the general background to the present invention, and should not
When being considered to recognize or imply in any form this information structure prior art well known to persons skilled in the art.
Summary of the invention
In order to overcome the deficiencies in the prior art, the invention provides a kind of microwave-assisted and prepare fresh-water fishes hemepeptide with membrane filtration
Method.
For solve above-mentioned technical problem, the present invention by the following technical solutions:
A kind of method that microwave-assisted and membrane filtration prepare fresh-water fishes hemepeptide, comprises the steps:
(1) preparation of anticoagulant fresh-water fishes blood: first put in container 1 by anticoagulant solution, then takes new fresh freshwater fish blood and puts
Enter in this container, stir, stand at low temperature, obtain anticoagulant fresh-water fishes blood；
(2) blood plasma, blood cell separate: anticoagulant fresh-water fishes blood upper strata is blood plasma, and lower floor is blood cell, is drawn onto by blood plasma with siphonage
In container 2；Remaining blood cell by centrifugation after, upper plasma is merged into freezing in container 2 standby, blood cell then chilled storage
To container 3 the most stand-by；
(3) blood plasma precipitates collects thrombinogen: after the blood plasma unfreezing in container 2, filter the White Flocculus that upper strata is floating
After, adjust pH to 5.1-5.2 with 2mol/L acetic acid again with after 2-8 times of normal saline dilution, stand at low temperature overnight, collects precipitation, i.e.
Obtain thrombinogen；
(4) prepared by blood coagulation crude enzyme liquid: be precipitated and dissolved in normal saline by the thrombinogen of collection, starts microwave device and adds
Heat insulation, to 37 DEG C, uses 1.0%CaCl2Solution, stirs 20-30min, activates thrombinogen, filtrate is collected by filtration, obtains blood coagulation
Enzyme crude enzyme liquid；
(5) preparation of thrombin: add cold saline in thrombin crude enzyme liquid, by ultrafiltration membrane treatment, be concentrated into former
After the l/20-1/25 of volume, collecting trapped fluid, this trapped fluid is refined thrombin liquid, by trapped fluid lyophilization, obtains powder
Shape thrombin；
(6) blood cell breaking cellular wall, haemolysis: add normal saline in container 3, obtain blood cell solution, be warming up to 10-15 DEG C, stir
Mix 20-30min；
(7) hyperglobulinemia subsection enzymolysis: regulate pH to 7.5 with 4.0moL/L NaOH, starts microwave device, by temperature liter
To 37-40 DEG C, add compound protease I by substrate quality 2-2.5%, after insulation 2h, then be warming up to 55-60 DEG C, be incubated 1.5-
2h；Regulation pH to 9.0, adds compound protease II by substrate quality 3-4.5%, restarts microwave extraction device, is incubated 2h, makes
Hyperglobulinemia is fully hydrolyzed；
(8) prepared by fresh-water fishes hemepeptide: after enzymolysis terminates, and is cooled to room temperature, degerming with the membrane filtration of 0.22um, spray dried
Dry, i.e. obtain fresh-water fishes hemepeptide powder.
As preferably, the anticoagulant solution described in step (1) is that anticoagulant trisodium citrate is dissolved to deionized water
Obtain, described in be the mass percent of anticoagulant solution be 0.5-5%.
As preferably, rotating speed centrifugal in step (2) is 3000-3500r/min.
As preferably, in step (4), the rotating speed of stirring is 500-800r/min.
As preferably, in step (5), the addition of cold saline is 8-10 times of thrombin crude enzyme liquid volume.
As preferably, in step (5), ultrafilter membrane is to retain the ultrafilter membrane that relative molecular weight is 6000.
As preferably, in step (6), the addition of normal saline is 8-10% for making blood cell protein content in solution.
As preferably, in step (6), the rotating speed of stirring is 1000-5000r/min.
As preferably, in step (7), compound protease I is alkalescence enzyme；Compound protease II is neutral enzymatic.
As preferably, the frequency of the microwave device described in step (4) and (7) is 2450mHZ, and power is 20KW.
Compared with prior art, there is advantages that
1. the method for the present invention improves the enzymolysis process of fresh-water fishes haemproteins and technology, by by microwave-assisted enzymolysis skill
Art preparation has bioactive oligopeptide, expands fresh water fin albumen application, turns waste into wealth, make from expansion of breeding fish merely
To multiple industry and the fields such as animal and fowl fodder, feed additive to human food, medicine, health product, product of undoubtedly prolongation being breeded fish
Industry chain, improves industry comprehensive benefit of breeding fish, it is achieved growth of agricultural efficiency increasing peasant income rural area is flourishing, to promoting-carry a road construction comprehensively
It is of great importance.
Microwave-assisted the most of the present invention and membrane filtration are prepared the method for fresh-water fishes hemepeptide and are had that yield is high, purity is high, solvent disappears
Consumption less, low cost and the advantage that is prone to industrialized production.
3. the whole technological process of the present invention is short, the time is short, and energy consumption is low, pollution less, does not use toxic reagent, contamination-free to arrange
Put, reach to clean productive target.
Accompanying drawing explanation
Fig. 1 is the process chart that microwave-assisted of the present invention and membrane filtration prepare fresh-water fishes hemepeptide.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is made further details of elaboration, but embodiments of the present invention are not
It is confined to the scope that embodiment represents.These embodiments are merely to illustrate the present invention, not for limiting the scope of the present invention.This
Outward, after reading present disclosure, those skilled in the art can various modifications may be made to the present invention, and these equivalent variations are same
Sample falls within appended claims limited range of the present invention.
Experimental technique used in following embodiment if no special instructions, is conventional method.Institute in following embodiment
The material of use, reagent etc., if no special instructions, the most commercially obtain.
Embodiment 1:
A kind of method that microwave-assisted and membrane filtration prepare fresh-water fishes hemepeptide, comprises the steps:
(1) preparation of anticoagulant fresh-water fishes blood: first put in container 1 by anticoagulant solution, then takes new fresh freshwater fish blood and puts
Enter in this container, stir, stand at low temperature, obtain anticoagulant fresh-water fishes blood；Described anticoagulant solution is by anticoagulant Fructus Citri Limoniae
Acid trisodium is dissolved to what deionized water obtained, described in be the mass percent of anticoagulant solution be 0.5%；
(2) blood plasma, blood cell separate: anticoagulant fresh-water fishes blood upper strata is blood plasma, and lower floor is blood cell, is drawn onto by blood plasma with siphonage
In container 2；Remaining blood cell by centrifugation after, upper plasma is merged into freezing in container 2 standby, blood cell then chilled storage
To container 3 the most stand-by；Centrifugal rotating speed is 3000r/min；
(3) blood plasma precipitates collects thrombinogen: after the blood plasma unfreezing in container 2, filter the White Flocculus that upper strata is floating
After, adjust pH to 5.1 with 2mol/L acetic acid again with after 2 times of normal saline dilutions, stand at low temperature overnight, is collected precipitation, is obtained blood coagulation
Proenzyme；
(4) prepared by blood coagulation crude enzyme liquid: be precipitated and dissolved in normal saline by the thrombinogen of collection, starts microwave device and adds
Heat insulation, to 37 DEG C, uses 1.0%CaCl2Solution, stirs 20min, activates thrombinogen, filtrate is collected by filtration, obtains thrombin
Crude enzyme liquid；The rotating speed of stirring is 500r/min；The frequency of described microwave device is 2450mHZ, and power is 20KW；
(5) preparation of thrombin: add cold saline in thrombin crude enzyme liquid, by ultrafiltration membrane treatment, be concentrated into former
After the l/20 of volume, collecting trapped fluid, this trapped fluid is refined thrombin liquid, by trapped fluid lyophilization, obtains powdery and coagulates
Hemase；The addition of cold saline is 8 times of thrombin crude enzyme liquid volume；Ultrafilter membrane is that to retain relative molecular weight be 6000
Ultrafilter membrane；
(6) blood cell breaking cellular wall, haemolysis: add normal saline in container 3, obtain blood cell solution, be warming up to 10 DEG C, stirring
20min；The rotating speed of stirring is 1000r/min；The addition of normal saline is 8% for making blood cell protein content in solution；
(7) hyperglobulinemia subsection enzymolysis: regulate pH to 7.5 with 4.0moL/L NaOH, starts microwave device, by temperature liter
To 37 DEG C, add compound protease I by substrate quality 2%, after insulation 2h, then be warming up to 55 DEG C, be incubated 1.5h；Regulation pH is extremely
9.0, add compound protease II by substrate quality 3%, restart microwave extraction device, be incubated 2h, make the abundant water of hyperglobulinemia
Solve；Compound protease I is alkalescence enzyme；Compound protease II is neutral enzymatic；The frequency of described microwave device is 2450mHZ, merit
Rate is 20KW；
(8) prepared by fresh-water fishes hemepeptide: after enzymolysis terminates, and is cooled to room temperature, degerming with the membrane filtration of 0.22um, spray dried
Dry, i.e. obtain fresh-water fishes hemepeptide powder.
Embodiment 2:
A kind of method that microwave-assisted and membrane filtration prepare fresh-water fishes hemepeptide, comprises the steps:
(1) preparation of anticoagulant fresh-water fishes blood: first put in container 1 by anticoagulant solution, then takes new fresh freshwater fish blood and puts
Enter in this container, stir, stand at low temperature, obtain anticoagulant fresh-water fishes blood；Described anticoagulant solution is by anticoagulant Fructus Citri Limoniae
Acid trisodium is dissolved to what deionized water obtained, described in be the mass percent of anticoagulant solution be 5%；
(2) blood plasma, blood cell separate: anticoagulant fresh-water fishes blood upper strata is blood plasma, and lower floor is blood cell, is drawn onto by blood plasma with siphonage
In container 2；Remaining blood cell by centrifugation after, upper plasma is merged into freezing in container 2 standby, blood cell then chilled storage
To container 3 the most stand-by；Centrifugal rotating speed is 3500r/min；
(3) blood plasma precipitates collects thrombinogen: after the blood plasma unfreezing in container 2, filter the White Flocculus that upper strata is floating
After, adjust pH to 5.2 with 2mol/L acetic acid again with after 8 times of normal saline dilutions, stand at low temperature overnight, is collected precipitation, is obtained blood coagulation
Proenzyme；
(4) prepared by blood coagulation crude enzyme liquid: be precipitated and dissolved in normal saline by the thrombinogen of collection, starts microwave device and adds
Heat insulation, to 37 DEG C, uses 1.0%CaCl2Solution, stirs 30min, activates thrombinogen, filtrate is collected by filtration, obtains thrombin
Crude enzyme liquid；The rotating speed of stirring is 800r/min；The frequency of described microwave device is 2450mHZ, and power is 20KW；
(5) preparation of thrombin: add cold saline in thrombin crude enzyme liquid, by ultrafiltration membrane treatment, be concentrated into former
Behind the 1/25 of volume, collecting trapped fluid, this trapped fluid is refined thrombin liquid, by trapped fluid lyophilization, obtains powdery and coagulates
Hemase；The addition of cold saline is 8-10 times of thrombin crude enzyme liquid volume；Ultrafilter membrane for retaining relative molecular weight is
The ultrafilter membrane of 6000；
(6) blood cell breaking cellular wall, haemolysis: add normal saline in container 3, obtain blood cell solution, be warming up to 15 DEG C, stirring
30min；The rotating speed of stirring is 5000r/min；The addition of normal saline is 10% for making blood cell protein content in solution；
(7) hyperglobulinemia subsection enzymolysis: regulate pH to 7.5 with 4.0moL/L NaOH, starts microwave device, by temperature liter
To 40 DEG C, add compound protease I by substrate quality 2.5%, after insulation 2h, then be warming up to 60 DEG C, be incubated 2h；Regulation pH is extremely
9.0, add compound protease II by substrate quality 4.5%, restart microwave extraction device, be incubated 2h, make hyperglobulinemia abundant
Hydrolysis；Compound protease I is alkalescence enzyme；Compound protease II is neutral enzymatic；The frequency of described microwave device is 2450mHZ,
Power is 20KW；
(8) prepared by fresh-water fishes hemepeptide: after enzymolysis terminates, and is cooled to room temperature, degerming with the membrane filtration of 0.22um, spray dried
Dry, i.e. obtain fresh-water fishes hemepeptide powder.
Embodiment 3:
A kind of method that microwave-assisted and membrane filtration prepare fresh-water fishes hemepeptide, comprises the steps:
(1) preparation of anticoagulant fresh-water fishes blood: first put in container 1 by anticoagulant solution, then takes new fresh freshwater fish blood and puts
Enter in this container, stir, stand at low temperature, obtain anticoagulant fresh-water fishes blood；Described anticoagulant solution is by anticoagulant Fructus Citri Limoniae
Acid trisodium is dissolved to what deionized water obtained, described in be the mass percent of anticoagulant solution be 3.0%；
(2) blood plasma, blood cell separate: anticoagulant fresh-water fishes blood upper strata is blood plasma, and lower floor is blood cell, is drawn onto by blood plasma with siphonage
In container 2；Remaining blood cell by centrifugation after, upper plasma is merged into freezing in container 2 standby, blood cell then chilled storage
To container 3 the most stand-by；Centrifugal rotating speed is 3200r/min；
(3) blood plasma precipitates collects thrombinogen: after the blood plasma unfreezing in container 2, filter the White Flocculus that upper strata is floating
After, adjust pH to 5.1 with 2mol/L acetic acid again with after 5 times of normal saline dilutions, stand at low temperature overnight, is collected precipitation, is obtained blood coagulation
Proenzyme；
(4) prepared by blood coagulation crude enzyme liquid: be precipitated and dissolved in normal saline by the thrombinogen of collection, starts microwave device and adds
Heat insulation, to 37 DEG C, uses 1.0%CaCl2Solution, stirs 25min, activates thrombinogen, filtrate is collected by filtration, obtains thrombin
Crude enzyme liquid；The rotating speed of stirring is 700r/min；The frequency of described microwave device is 2450mHZ, and power is 20KW；
(5) preparation of thrombin: add cold saline in thrombin crude enzyme liquid, by ultrafiltration membrane treatment, be concentrated into former
After the l/22 of volume, collecting trapped fluid, this trapped fluid is refined thrombin liquid, by trapped fluid lyophilization, obtains powdery and coagulates
Hemase；The addition of cold saline is 9 times of thrombin crude enzyme liquid volume；Ultrafilter membrane is that to retain relative molecular weight be 6000
Ultrafilter membrane；
(6) blood cell breaking cellular wall, haemolysis: add normal saline in container 3, obtain blood cell solution, be warming up to 12 DEG C, stirring
25min；The rotating speed of stirring is 3000r/min；The addition of normal saline is 9% for making blood cell protein content in solution；
(7) hyperglobulinemia subsection enzymolysis: regulate pH to 7.5 with 4.0moL/L NaOH, starts microwave device, by temperature liter
To 38 DEG C, add compound protease I by substrate quality 2.2%, after insulation 2h, then be warming up to 58 DEG C, be incubated 1.8h；Regulation pH is extremely
9.0, add compound protease II by substrate quality 4.0%, restart microwave extraction device, be incubated 2h, make hyperglobulinemia abundant
Hydrolysis；Compound protease I is alkalescence enzyme；Compound protease II is neutral enzymatic；The frequency of described microwave device is 2450mHZ,
Power is 20KW；
(8) prepared by fresh-water fishes hemepeptide: after enzymolysis terminates, and is cooled to room temperature, degerming with the membrane filtration of 0.22um, spray dried
Dry, i.e. obtain fresh-water fishes hemepeptide powder.
Comparison: with reference to the method in Application No. 201510162420.4 patent.
The extracting method of a kind of loach fish corner material collagen protein, it is characterised in that extraction step is as follows:
In a, loach fish, fish skin, fishbone and fin are raw material, use special equipment to pulverize raw material；To the raw material after pulverizing
It is watered, is allowed to become serosity；Acidolysis implemented by b, material acid to addition in serosity；It is blended into the water and pulverizing raw material pulverized in raw material
Ratio be 3-8:1；C, to after acidolysis serosity implement ultrasonic Treatment；The time of ultrasonic Treatment is 2h；The time of extraction
For 4h；D, serosity implement extraction after being ultrasonically treated；Acid material is citric acid；E, filtration；Filter and use filter, will
Upper strata oil layer and deposit in serosity are implemented to filter, and form the acid solution containing collagen protein；F, desalination: be by containing collagen
The acid solution of albumen, respectively by purpose ceramic-film filter and NF membrane desalination；G, freezing, cryogenic temperature is subzero 30 DEG C；H, dry
Dry, the temperature being dried is 8-15 DEG C.
The beneficial effect of table 1 extracting method of the present invention
| Group | Enzymolysis time (h) | Conversion ratio (%) |
| Embodiment 1 | 0.59 | 81.20 |
| Embodiment 2 | 0.65 | 83.4 |
| Embodiment 3 | 0.65 | 85.2 |
| Comparison | 6 | 61.2 |
As known from Table 1, compared with contrast method, present invention microwave-assisted and membrane filtration technique, it is not required to solvent, soda acid
Consumption is few, and enzymolysis time is short, and conversion ratio is high, and prepares the polypeptide more preferable condition of creation for next step.
The aforementioned description to the specific illustrative embodiment of the present invention illustrates that and the purpose of illustration.These describe
It is not wishing to limit the invention to disclosed precise forms, and it will be apparent that according to above-mentioned teaching, can much change
And change.The purpose selected exemplary embodiment and describe is to explain that the certain principles of the present invention and reality thereof should
With so that those skilled in the art be capable of and utilize the present invention various different exemplary and
Various different selections and change.The scope of the present invention is intended to be limited by claims and equivalents thereof.
Claims (10)
1. the method that a microwave-assisted prepares fresh-water fishes hemepeptide with membrane filtration, it is characterised in that comprise the steps:
(1) preparation of anticoagulant fresh-water fishes blood: first put in container 1 by anticoagulant solution, then takes new fresh freshwater fish blood and puts into this
In container, stir, stand at low temperature, obtain anticoagulant fresh-water fishes blood；
(2) blood plasma, blood cell separate: anticoagulant fresh-water fishes blood upper strata is blood plasma, and lower floor is blood cell, with siphonage, blood plasma is drawn onto container
In 2；Remaining blood cell by centrifugation after, upper plasma is merged into freezing in container 2 standby, blood cell then chilled storage to hold
In device 3 the most stand-by；
(3) blood plasma precipitates collects thrombinogen: after the blood plasma unfreezing in container 2, after filtering the White Flocculus that upper strata is floating, uses
Adjusting pH to 5.1-5.2 with 2mol/L acetic acid again after 2-8 times of normal saline dilution, stand at low temperature overnight, is collected precipitation, is obtained blood coagulation
Proenzyme；
(4) prepared by blood coagulation crude enzyme liquid: be precipitated and dissolved in normal saline by the thrombinogen of collection, starts microwave device heating and protects
Temperature, to 37 DEG C, uses 1.0%CaCl2Solution, stirs 20-30min, activates thrombinogen, filtrate is collected by filtration, obtains thrombin thick
Enzyme liquid；
(5) preparation of thrombin: add cold saline in thrombin crude enzyme liquid, by ultrafiltration membrane treatment, be concentrated into original volume
L/20-1/25 after, collect trapped fluid, this trapped fluid is refined thrombin liquid, by trapped fluid lyophilization, obtain powdery coagulate
Hemase；
(6) blood cell breaking cellular wall, haemolysis: add normal saline in container 3, obtain blood cell solution, be warming up to 10-15 DEG C, stir 20-
30min；
(7) hyperglobulinemia subsection enzymolysis: regulate pH to 7.5 with 4.0moL/L NaOH, starts microwave device, temperature is risen to 37-
40 DEG C, add compound protease I by substrate quality 2-2.5%, after insulation 2h, then be warming up to 55-60 DEG C, be incubated 1.5-2h；Adjust
Joint pH to 9.0, adds compound protease II by substrate quality 3-4.5%, restarts microwave extraction device, is incubated 2h, makes blood cell
Albumen is fully hydrolyzed；
(8) prepared by fresh-water fishes hemepeptide: after enzymolysis terminates, and is cooled to room temperature, degerming with the membrane filtration of 0.22um, is spray-dried,
I.e. obtain fresh-water fishes hemepeptide powder.
The method that microwave-assisted the most according to claim 1 and membrane filtration prepare fresh-water fishes hemepeptide, it is characterised in that step
(1) anticoagulant trisodium citrate is dissolved to deionized water and obtains by the anticoagulant solution described in, described in be that anticoagulant is molten
The mass percent of liquid is 0.5-5%.
The method that microwave-assisted the most according to claim 1 and membrane filtration prepare fresh-water fishes hemepeptide, it is characterised in that step
(2) rotating speed centrifugal in is 3000-3500r/min.
The method that microwave-assisted the most according to claim 1 and membrane filtration prepare fresh-water fishes hemepeptide, it is characterised in that step
(4) in, the rotating speed of stirring is 500-800r/min.
The method that microwave-assisted the most according to claim 1 and membrane filtration prepare fresh-water fishes hemepeptide, it is characterised in that step
(5) in, the addition of cold saline is 8-10 times of thrombin crude enzyme liquid volume.
The method that microwave-assisted the most according to claim 1 and membrane filtration prepare fresh-water fishes hemepeptide, it is characterised in that step
(5) in, ultrafilter membrane is to retain the ultrafilter membrane that relative molecular weight is 6000.
The method that microwave-assisted the most according to claim 1 and membrane filtration prepare fresh-water fishes hemepeptide, it is characterised in that step
(6) in, the addition of normal saline is 8-10% for making blood cell protein content in solution.
The method that microwave-assisted the most according to claim 1 and membrane filtration prepare fresh-water fishes hemepeptide, it is characterised in that step
(6) in, the rotating speed of stirring is 1000-5000r/min.
The method that microwave-assisted the most according to claim 1 and membrane filtration prepare fresh-water fishes hemepeptide, it is characterised in that step
(7) in, compound protease I is alkalescence enzyme；Compound protease II is neutral enzymatic.
The method that microwave-assisted the most according to claim 1 and membrane filtration prepare fresh-water fishes hemepeptide, it is characterised in that step
Suddenly the frequency of the microwave device described in (4) and (7) is 2450mHZ, and power is 20KW.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610652842.4A CN106086140A (en) | 2016-08-10 | 2016-08-10 | A kind of method that microwave-assisted and membrane filtration prepare fresh-water fishes hemepeptide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610652842.4A CN106086140A (en) | 2016-08-10 | 2016-08-10 | A kind of method that microwave-assisted and membrane filtration prepare fresh-water fishes hemepeptide |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101979532A (en) * | 2010-10-13 | 2011-02-23 | 江苏省江大绿康生物工程技术研究有限公司 | Method for comprehensively using pig blood |
| CN102146427A (en) * | 2010-12-30 | 2011-08-10 | 吉林大学 | Method for preparing swine blood active peptide by microwave-accelerated enzymolysis |
| CN103952458A (en) * | 2014-04-30 | 2014-07-30 | 桂林军供生化技术开发有限公司 | Method for preparing active peptides in duck blood through microwave-assisted enzymolysis |
| CN105331665A (en) * | 2015-12-09 | 2016-02-17 | 梁尚文 | Method for preparing brain polypeptide and brain small-molecule peptide by means of pig brain protein through enzymolysis |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101979532A (en) * | 2010-10-13 | 2011-02-23 | 江苏省江大绿康生物工程技术研究有限公司 | Method for comprehensively using pig blood |
| CN102146427A (en) * | 2010-12-30 | 2011-08-10 | 吉林大学 | Method for preparing swine blood active peptide by microwave-accelerated enzymolysis |
| CN103952458A (en) * | 2014-04-30 | 2014-07-30 | 桂林军供生化技术开发有限公司 | Method for preparing active peptides in duck blood through microwave-assisted enzymolysis |
| CN105331665A (en) * | 2015-12-09 | 2016-02-17 | 梁尚文 | Method for preparing brain polypeptide and brain small-molecule peptide by means of pig brain protein through enzymolysis |
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Application publication date: 20161109 |