CN106520882A - Production method of extracting small-peptide ferroheme from animal blood - Google Patents
Production method of extracting small-peptide ferroheme from animal blood Download PDFInfo
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- CN106520882A CN106520882A CN201611134138.6A CN201611134138A CN106520882A CN 106520882 A CN106520882 A CN 106520882A CN 201611134138 A CN201611134138 A CN 201611134138A CN 106520882 A CN106520882 A CN 106520882A
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- C12P21/00—Preparation of peptides or proteins
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- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
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Abstract
The invention discloses a production method for extracting small-peptide ferroheme from animal blood, and relates to the field of the production and the preparation of the small-peptide ferroheme. The method comprises the following steps: (1) collecting blood; (2) conducting anticoagulation; (3) conducting standing by and separation; (4) conducting centrifugal separation; (5) conducting hemolysis; (6) conducting ultrasonic disruption; (7) conducting filtration; (8) conducting enzymatic hydrolysis; (9) resisting oxidation and promoting absorption; (10) conducting spray-drying; (11) conducting separation and crystallization; and (12) conducting mixing and shaping. The production method has the advantages of being reasonable in production method and high in resource utilization rate, and the prepared small-peptide ferroheme is good in quality and taste and the small-peptide ferroheme can be easily absorbed by human body.
Description
Technical field
The present invention relates to the production preparation field of small peptide heme, more particularly to one kind is from animal blood extraction small peptide
The production method of heme.
Background technology
Iron deficiency anemia (iron deficiency anemia, abbreviation IDA) is global nutrition problem, is serious danger
The common nutritional disease of victimization class, particularly child.The whole world there are about 5～600,000,000 people and suffer from IDA according to statistics, and China's trophism is lean
Have 95.65% in blood patient for IDA.Calendar year 2001, the Ministry of Education of the state was adjusted to the students in middle and primary schools of 7～14 years old age level of China
Look into, it is found that the student's ratio for suffering from IDA is up to 27%, it was demonstrated that China's students in middle and primary schools' constitution status make us disturbing.It has been investigated that,
During IDA can cause the reason for series of symptoms such as slow upgrowth and development of children, mental retardation, immunity degradation, morbidity to be food
Iron content is not enough, and the source great majority of ferrum are vegetable foods, and absorption rate is low.
Small peptide heme is natural novel ecological iron preparation, is for the heretofore known ideal anti-anemia action of the mankind
Medicine, has：⑴Fe2+Content is high, good stability；(2) it is in good taste；(3) splendid absorption rate.Small peptide heme can have
Effect preventing and treating IDA (iron deficiency anemia iron deficiency anemia), the total effective rate for treating IDA reach 96.2%, significantly
Higher than the iron preparation of other any state types.Small peptide heme is widely used as the hardening agent of functional food.
The content of the invention
For the deficiencies in the prior art, it is to provide a kind of from animal blood extraction small peptide that the purpose of the present invention is
The production method of heme, rationally, resource utilization is high for its production method, and the quality of the small peptide heme of preparation is good
And mouthfeel is good.
For achieving the above object, the present invention is achieved by the following technical programs：
A kind of production method for extracting small peptide heme from animal blood, the method comprise the steps：
(1) take a blood sample：In slaughter hall, animal blood is collected rapidly with tube type vacuum blood sampling knife, the blood collected is deposited
It is placed in collecting bag or resealable container；
(2) anticoagulant：Anticoagulant and ascorbic acid are pressed before blood sampling the 2%-15% and 0.045%- of blood sampling volume respectively
0.048% is added in collecting bag or resealable container；
(3) standing separation：After stirring and evenly mixing, stand 100-150 minutes, with siphonage by upper strata show ruddy blood plasma with
The wine-colored hemocyte of lower floor is separated；
(4) centrifugation：The hemocyte liquid that standing separation is gone out, is centrifuged 10 minutes in the centrifuge of 4500r/m, removes
Upper plasma, further concentrates erythrocyte；
(5) haemolysis：The erythrocyte of Jing centrifugal concentratings is put into into freezing 2-4 hours in -20 DEG C of freezer, is taken out, will freezing
Hemocyte in 50 DEG C -54 DEG C of tepidarium haemolysis, 3-4 time repeatedly；
(6) ultrasonic disruption：Hemolysate is put in ultrasonic unit carries out broken 8-12min, while utilizing Glass rod
It is stirred, obtains mixed liquor；
(7) filter：By mixed liquor by hollow fiber membrane filter, membrane component stromatin is removed, purer blood is obtained
Red eggs white liquor；
(8) digest：Digested with compound protease substep gradual control in an inert atmosphere, obtain small peptide heme, adopt
With trypsin, flavor protease and subtilisin substep controlled enzymatic hydrolysis, tryptic technological parameter is added
PH7.0-7.5, temperature 45-48 DEG C, time 2-3 hour, add flavor protease technological parameter be PH6.0-6.5, temperature 52-
54 DEG C, time 3-4 hour, add subtilisin technological parameter be PH6.5-7.0, temperature 49-51 DEG C, time 3-4
Hour, after the completion of enzymolysis, enzyme denaturing obtains digesting solution；
(9) antioxygen promotees to absorb：Ascorbic acid and phosphopeptide caseinate composition mixture are added in enzymolysis solution；
(10) it is spray-dried：Mixture is dried using spray drying method, obtains small peptide heme；
(11) fractional crystallization：The blood plasma mixing that centrifugation goes out in the blood plasma that standing separation in step 3 is gone out and step 4,
By the blood plasma vacuum distillation of mixing to dry, by the residue (g) being evaporated：Hydrochloric acid (ml)=1:7 mass volume ratio, toward what is be evaporated
Add concentration for the hydrochloric acid solution of 21-25% in residue, be warming up to 80-100 DEG C, be stirred at reflux 2-5h, be cooled to 5-10 DEG C
2-8h is stood still for crystals, is filtered, is obtained small peptide heme crystal, be dried after washing with water, obtain small peptide heme；
(12) mixed-forming：By obtained small peptide ferrous iron blood in obtained small peptide heme in step 10 and step 11
Red pigment mixes, and using milling device pulverizing, is then sieved using vibrosieve, is not come back to by the powder particle of sieve aperture
Pulverizing operation is carried out in flour mill, is powder small peptide heme finished product by the powder of sieve aperture.
Further, the anticoagulant in the step 2 in methods described is one or two the mixing in EDTA and heparin sodium
Thing.
Further, trypsin, flavor protease and subtilisin and blood red egg in the step 8 in methods described
The mass ratio of white liquor is respectively 8:100、9:100、3:100.
Further, ascorbic acid and phosphopeptide caseinate and the quality score for digesting solution in the step 9 in methods described
Wei 3:100 and 1:50.
In sum, it is an advantage of the invention that：The production method that small peptide heme is extracted from animal blood is reasonable, leads to
The extraction ratio that the method can effectively improve little too heme is crossed, resource utilization is high, and the small peptide ferrous iron of preparation is blood red
Plain quality is good, mouthfeel is good, and is easily absorbed by the body.
Specific embodiment
Embodiment
A kind of production method for extracting small peptide heme from animal blood, the method comprise the steps：
(1) take a blood sample：In slaughter hall, animal blood is collected rapidly with tube type vacuum blood sampling knife, the blood collected is deposited
It is placed in collecting bag or resealable container；
(2) anticoagulant：Anticoagulant and ascorbic acid are pressed before blood sampling the 2%-15% and 0.045%- of blood sampling volume respectively
0.048% is added in collecting bag or resealable container；
(3) standing separation：After stirring and evenly mixing, stand 100-150 minutes, with siphonage by upper strata show ruddy blood plasma with
The wine-colored hemocyte of lower floor is separated；
(4) centrifugation：The hemocyte liquid that standing separation is gone out, is centrifuged 10 minutes in the centrifuge of 4500r/m, removes
Upper plasma, further concentrates erythrocyte；
(5) haemolysis：The erythrocyte of Jing centrifugal concentratings is put into into freezing 2-4 hours in -20 DEG C of freezer, is taken out, will freezing
Hemocyte in 50 DEG C -54 DEG C of tepidarium haemolysis, 3-4 time repeatedly；
(6) ultrasonic disruption：Hemolysate is put in ultrasonic unit carries out broken 8-12min, while utilizing Glass rod
It is stirred, obtains mixed liquor；
(7) filter：By mixed liquor by hollow fiber membrane filter, membrane component stromatin is removed, purer blood is obtained
Red eggs white liquor；
(8) digest：Digested with compound protease substep gradual control in an inert atmosphere, obtain small peptide heme, adopt
With trypsin, flavor protease and subtilisin substep controlled enzymatic hydrolysis, tryptic technological parameter is added
PH7.0-7.5, temperature 45-48 DEG C, time 2-3 hour, add flavor protease technological parameter be PH6.0-6.5, temperature 52-
54 DEG C, time 3-4 hour, add subtilisin technological parameter be PH6.5-7.0, temperature 49-51 DEG C, time 3-4
Hour, after the completion of enzymolysis, enzyme denaturing obtains digesting solution；
(9) antioxygen promotees to absorb：Ascorbic acid and phosphopeptide caseinate composition mixture are added in enzymolysis solution；
(10) it is spray-dried：Mixture is dried using spray drying method, obtains small peptide heme；
(11) fractional crystallization：The blood plasma mixing that centrifugation goes out in the blood plasma that standing separation in step 3 is gone out and step 4,
By the blood plasma vacuum distillation of mixing to dry, by the residue (g) being evaporated：Hydrochloric acid (ml)=1:7 mass volume ratio, toward what is be evaporated
Add concentration for the hydrochloric acid solution of 21-25% in residue, be warming up to 80-100 DEG C, be stirred at reflux 2-5h, be cooled to 5-10 DEG C
2-8h is stood still for crystals, is filtered, is obtained small peptide heme crystal, be dried after washing with water, obtain small peptide heme；
(12) mixed-forming：By obtained small peptide ferrous iron blood in obtained small peptide heme in step 10 and step 11
Red pigment mixes, and using milling device pulverizing, is then sieved using vibrosieve, is not come back to by the powder particle of sieve aperture
Pulverizing operation is carried out in flour mill, is powder small peptide heme finished product by the powder of sieve aperture.
Further, the anticoagulant in the step 2 in methods described is one or two the mixing in EDTA and heparin sodium
Thing.
Further, trypsin, flavor protease and subtilisin and blood red egg in the step 8 in methods described
The mass ratio of white liquor is respectively 8:100、9:100、3:100.
Further, ascorbic acid and phosphopeptide caseinate and the quality score for digesting solution in the step 9 in methods described
Wei 3:100 and 1:50.
Should be from the production method step of animal blood extraction small peptide heme rationally, by fractional crystallization to isolating
Blood plasma carry out the extraction of secondary small peptide heme, effectively improve the extraction ratio of small peptide heme so that money
Source utilization rate gets a promotion；Increase ascorbic acid in anticoagulant and antioxygen promote absorption process simultaneously, in anti-coagulation process, add anticoagulant
Agent can effectively ensure that the quality of obtained product in the quality and production process of raw material, so as to little for what is prepared from material
The quality of peptide heme provides guarantee；Add EDTA in antioxygen promotees absorption process and can promote small peptide heme quilt
Human small intestine absorbs；The crushing effect of erythrocyte is effectively raised using two steps of haemolysis and ultrasonic disruption so that work
Lifted as efficiency, improved the extraction ratio of haemoglobin liquid；Trypsin, flavor protease and hay bar is arrived used in enzymolysis
Mycoproteinase so that haemoglobin liquid fully can be digested, lifts the mouthfeel of the small peptide heme of production.
For a person skilled in the art, other can be made various corresponding according to above technical scheme and design
Change and deformation, and all these change and deformation should all belong within the protection domain of the claims in the present invention.
Claims (4)
1. it is a kind of from animal blood extract small peptide heme production method, it is characterised in that：The method includes following step
Suddenly：
(1) take a blood sample：In slaughter hall, animal blood is collected rapidly with tube type vacuum blood sampling knife, the blood collected is stored in
In collecting bag or resealable container；
(2) anticoagulant：Anticoagulant and ascorbic acid are added by the 2%-15% and 0.045%-0.048% of blood sampling volume respectively before blood sampling
To in collecting bag or resealable container；
(3) standing separation：After stirring and evenly mixing, 100-150 minutes are stood, upper strata is shown into ruddy blood plasma and lower floor with siphonage
Wine-colored hemocyte is separated；
(4) centrifugation：The hemocyte liquid that standing separation is gone out, is centrifuged 10 minutes in the centrifuge of 4500r/m, removes upper strata
Blood plasma, further concentrates erythrocyte；
(5) haemolysis：The erythrocyte of Jing centrifugal concentratings is put into into freezing 2-4 hours in -20 DEG C of freezer, is taken out, by the blood of freezing
Cell haemolysis in 50 DEG C -54 DEG C of tepidarium, 3-4 time repeatedly；
(6) ultrasonic disruption：Hemolysate is put in ultrasonic unit carries out broken 8-12min, while being carried out using Glass rod
Stirring, obtains mixed liquor；
(7) filter：By mixed liquor by hollow fiber membrane filter, membrane component stromatin is removed, purer blood red egg is obtained
White liquor；
(8) digest：Digested with compound protease substep gradual control in an inert atmosphere, obtain small peptide heme, using pancreas
Protease, flavor protease and subtilisin substep controlled enzymatic hydrolysis, add tryptic technological parameter PH7.0-
7.5, temperature 45-48 DEG C, time 2-3 hour, add flavor protease technological parameter be PH6.0-6.5, temperature 52-54 DEG C,
Time 3-4 hour, add subtilisin technological parameter be PH6.5-7.0, temperature 49-51 DEG C, time 3-4 hour,
After the completion of enzymolysis, enzyme denaturing obtains digesting solution；
(9) antioxygen promotees to absorb：Ascorbic acid and phosphopeptide caseinate composition mixture are added in enzymolysis solution；
(10) it is spray-dried：Mixture is dried using spray drying method, obtains small peptide heme；
(11) fractional crystallization：The blood plasma mixing that centrifugation goes out in the blood plasma that standing separation in step 3 is gone out and step 4, will be mixed
The blood plasma vacuum distillation of conjunction to dry, by the residue (g) being evaporated：Hydrochloric acid (ml)=1:7 mass volume ratio, toward the residual being evaporated
Add concentration for the hydrochloric acid solution of 21-25% in thing, be warming up to 80-100 DEG C, be stirred at reflux 2-5h, be cooled to 5-10 DEG C of standing
Crystallization 2-8h, filters, obtains small peptide heme crystal, is dried, obtains small peptide heme after washing with water；
(12) mixed-forming：By obtained small peptide heme in obtained small peptide heme in step 10 and step 11
Mixing, using milling device pulverizing, is then sieved using vibrosieve, is not come back to pulverizing by the powder particle of sieve aperture
Pulverizing operation is carried out in machine, is powder small peptide heme finished product by the powder of sieve aperture.
2. the production method of small peptide heme is extracted from animal blood according to claim 1, it is characterised in that：It is described
Anticoagulant in step 2 in method is one or two the mixture in EDTA and heparin sodium.
3. the production method of small peptide heme is extracted from animal blood according to claim 1, it is characterised in that：It is described
In step 8 in method, the mass ratio of trypsin, flavor protease and subtilisin and haemoglobin liquid is respectively
8:100、9:100、3:100。
4. the production method of small peptide heme is extracted from animal blood according to claim 1, it is characterised in that：It is described
In step 9 in method, ascorbic acid and phosphopeptide caseinate are respectively 3 with the mass ratio of enzymolysis solution:100 and 1:50.
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Cited By (4)
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| CN110100944A (en) * | 2019-04-04 | 2019-08-09 | 中国农业科学院农产品加工研究所 | Preparation method rich in active ferrous iron ion animal blood oligopeptides |
| CN111018867A (en) * | 2019-12-12 | 2020-04-17 | 山东立菲生物产业有限公司 | Method for quickly extracting heme from hemoglobin |
| CN111387402A (en) * | 2020-03-04 | 2020-07-10 | 合肥工业大学 | Heme peptide protective agent and using method thereof |
| CN115010803A (en) * | 2021-03-03 | 2022-09-06 | 内蒙古天奇生物科技有限公司 | Preparation of hemoglobin polypeptide rich in heme iron |
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Application publication date: 20170322 |