CN101755997B - Sheep heme peptide and preparation method thereof - Google Patents
Sheep heme peptide and preparation method thereof Download PDFInfo
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- CN101755997B CN101755997B CN2010100009366A CN201010000936A CN101755997B CN 101755997 B CN101755997 B CN 101755997B CN 2010100009366 A CN2010100009366 A CN 2010100009366A CN 201010000936 A CN201010000936 A CN 201010000936A CN 101755997 B CN101755997 B CN 101755997B
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Abstract
The invention discloses a sheep heme peptide and a preparation method thereof, and belongs to the field of food additives. The preparation method mainly comprises the following steps: adding trisodium citrate to sheep blood, then centrifuging, and collecting a sheep hemoglobin solution obtained by separation; then placing in a sealed container to be vacuumized, and removing the smell of mutton; then adding compound protease composed of papain and neutral protease to be digested, centrifuging and collecting precipitation; then washing a sediment with distilled water, centrifuging to take the sediment; and finally, obtaining the sheep heme peptide by freezing and drying. The heme peptide of the invention has no smell of mutton and side effects, and has high safety; the recovery rate of heme can reach up to 95 percent above, and the heme content can reach up to 28 to 32%; and the preparation method of the invention is simple and easy and is low in cost.
Description
Technical field
The invention belongs to functional food and food additives field, be specifically related to a kind of sheep heme peptide and preparation method thereof.
Background technology
Animal blood protein Compositional balance contains 8 kinds of essential amino acids of needed by human body, is the high-quality protein source, has higher biological value, contains abundant organic iron simultaneously.Discharged or abandoned with the form of sewage but animal blood, particularly sheep blood are most of at present, cause a large amount of valuable nutritional resources to run off, return environment and caused severe contamination.
Heme iron is the red pigments in higher mammal blood, the muscle, and the porphines (Porphin) that its basic structure composition is made up of the pyrroles forms porphyrin (Porphyrin) after being with side chain, is heme iron with the ferrous formation ferriporphyrin of a part compound again.In hemoglobin and myoglobins, heme iron accounts for 3.8%, and heme iron content is about 0.5% in the whole blood.Heme iron is one of staple product of animal blood deep processing.The production method of ferroheme mainly contain the sodium acetate method (week light should the grade. medicine biotechnology, 2002,9 (2): 103-104; ), the way of distillation and carboxymethyl cellulose method.
(application number is: 200910058656.8) (application number is: 200710031826.4) disclose relevant pig blood peptide native or heme peptide and preparation method thereof with " preparation method of collapse due to massive hemorrhage red pigment porcine hemoglobin zymolyte and heme peptide " in patent application " hydrolyzation of pig blood with complex enzyme method for producing blood peptide native "; But up to the present; Also do not find the report of relevant sheep heme peptide; Mainly be because sheep blood has smell of mutton, be difficult to use as food additives.
Summary of the invention
The sheep heme peptide that the purpose of this invention is to provide a kind of without smelling of mutton.Sheep heme peptide of the present invention does not have bad local flavor, can directly make an addition to food; The ferroheme rate of recovery of the present invention is high, and content of hemachrome is high.
Another purpose of the present invention provides the preparation method of above-mentioned sheep heme peptide.
Realize that technical scheme of the present invention is following:
A kind of sheep heme peptide of the present invention prepares according to following method:
(1) preliminary treatment: the percentage by weight according to 0.3～0.5% adds trisodium citrate in sheep blood; Stir; At the centrifugal 6～10min of 4000～6000g, be separated into two layers of solution then, the upper strata is the sheep blood plasma protein solution; Lower floor is the sheep hemoglobin solutions, collects the sheep hemoglobin solutions of lower floor;
(2) gained sheep hemoglobin solutions is placed closed container, bleed with vavuum pump then, when vacuum reaches 0.07～0.09 MPa, be heated to 60～70 ℃ again and bled 3～5 minutes, obtain the sheep hemoglobin solutions of without smelling of mutton;
(3) the sheep hemoglobin solutions of step (2) gained is diluted to concentration is 15～20% to water; Regulating its pH is 6.5～7.5; According to mass ratio between compound protease and the hemoglobin is that 1: 50～100 ratio adds compound protease in the sheep hemoglobin solutions, is the reaction 4～7h that is hydrolyzed under 6.5～7.5 conditions at 40～45 ℃, pH then, again at 85～90 ℃ of insulation 2～4min enzymes that go out down; Then be cooled to 20 ℃; At the centrifugal 5～10min of 5000～6000g, collecting precipitation gets sediment;
(4) with the sediment of distilled water washing step (3) gained of 1～2 times of weight, again at the centrifugal 5min～10min of 4000g～5000g, taking precipitate; Press aforesaid operations repeated washing sediment 2～3 times; Freeze drying gets the sheep heme peptide.
Compound protease described in the above-mentioned sheep heme peptide step (3) is made up of papain and neutral proteinase, and the ratio of living of the enzyme between them is 1: 1.
Sheep blood described in the above-mentioned sheep heme peptide step (1) can be fresh sheep blood.
PH value described in the above-mentioned sheep heme peptide step (3) is regulated with the hydrochloric acid or the sodium hydroxide solution of 1 molar concentration.
Described papain and neutral proteinase all can be bought in market.
The preparation method of above-mentioned sheep heme peptide comprises the steps:
(1) preliminary treatment: the percentage by weight according to 0.3～0.5% adds trisodium citrate in sheep blood; Stir; At the centrifugal 6～10min of 4000～6000g, be separated into two layers of solution then, the upper strata is the sheep blood plasma protein solution; Lower floor is the sheep hemoglobin solutions, collects the sheep hemoglobin solutions of lower floor;
(2) gained sheep hemoglobin solutions is placed closed container, bleed with vavuum pump then, when vacuum reaches 0.07～0.09 MPa, be heated to 60～70 ℃ again and bled 3～5 minutes, obtain the sheep hemoglobin solutions of without smelling of mutton;
(3) the sheep hemoglobin solutions of step (2) gained is diluted to concentration is 15～20% to water, and regulating pH is 6.5～7.5, is that 1: 50～100 ratio adds compound protease in the sheep hemoglobin solutions according to mass ratio between compound protease and the hemoglobin; Reaction 4～7h then is hydrolyzed under 40～45 ℃, pH6.5～7.5 conditions; At 85～90 ℃ of following insulation 2～4min enzymes that go out, then be cooled to 20 ℃, again at the centrifugal 5～10min of 5000～6000g; Collecting precipitation gets sediment;
(4) with the sediment of distilled water washing step (3) gained of 1～2 times of weight, again at the centrifugal 5min～10min of 4000g～5000g, taking precipitate; Press aforesaid operations repeated washing sediment 2～3 times; Freeze drying gets the sheep heme peptide.
Compound protease described in above-mentioned preparation method's step (3) is made up of papain and neutral proteinase, and the ratio of living of the enzyme between them is 1: 1.
Sheep blood described in above-mentioned preparation method's step (1) can be fresh sheep blood.
PH value described in above-mentioned preparation method's step (3) is regulated with the hydrochloric acid or the sodium hydroxide solution of 1 molar concentration.
Described papain and neutral proteinase all can be bought on market.
Sheep heme peptide of the present invention can be used as functional food or food additives make an addition in the food.
The advantage that the present invention has: (1), sheep ferroheme without smelling of mutton of the present invention, adopt the vacuum suction method, can remove wherein 95% smell of mutton, its local flavor can be accepted by the consumer.(2), the present invention adopt food-grade albumen enzyme (papain and neutral proteinase) through under temperate condition enzymolysis prepare blood sheep red pigment peptide, it is safe, has no side effect.(3), sheep red pigment peptide of the present invention, absorptivity is high, has no adverse reaction after edible.(4) the ferroheme rate of recovery of the present invention is high, can reach more than 95%, and content of hemachrome is high, can reach 28～32%.(4), preparation method of the present invention is simple and easy to do, cost is low.
The specific embodiment
The preparation of embodiment 1 sheep heme peptide
Carry out according to following method:
(1), in the 500g sheep blood, to add concentration be 20% citric acid three sodium solution 10g, stir, then at the centrifugal 6min of 5000g; Separate and obtain two layers of solution; The upper strata is the sheep blood plasma protein solution, and lower floor is the sheep hemoglobin solutions, collects sheep hemoglobin solutions (325g).
(2), the gained hemoglobin solutions is placed airtight container, bleed with vavuum pump then, when vacuum reaches 0.085 MPa, be heated to 65 ℃ again, bled 3 minutes, obtain the sheep hemoglobin solutions of without smelling of mutton.
(3), step (2) gained sheep hemoglobin solutions is diluted to concentration is 15% to water; Regulating pH is 6.8; Adding compound protease (ratio that papain (productions of Nanning Jie Woli biological products Co., Ltd) and neutral proteinase (the magnificent prosperous and powerful Bioisystech Co., Ltd in east, Beijing produces) were lived 1: 1 by enzyme) 0.8g, is the reaction 5h that is hydrolyzed under 6.8 conditions at 42 ℃, pH, under 90 ℃, is incubated 2 minutes then; Be cooled to 20 ℃; At the centrifugal 8min of 5500g, collecting precipitation obtains sediment 15g;
(4), with 15g distilled water washing step (3) gained sediment, at 4500g centrifugal 9 minutes, taking precipitate; Repeat above-mentioned washing precipitate operation 3 times; With gained sediment freeze drying (FD-1PF, development in science and technology Co., Ltd is helped in moral sky, Beijing) 12 hours, get sheep heme peptide 7g again.
Adopt the content of iron in the potassium rhodanide colorimetric method for determining heme peptide, the content of iron is 7.2% in the heme peptide as a result.
The preparation of embodiment 2 sheep heme peptides
(1), in the 1000g sheep blood, adding concentration is 20% citric acid three sodium solution 18g; Stir, then at 5000g centrifugal (TGL-16A centrifuge, the ordinary instrument and meter in Changsha Co., Ltd) 7min; Separate and obtain two layers of solution; Lower floor is sheep hemoglobin solutions (660g), and the upper strata is the sheep blood plasma protein solution, collects the sheep hemoglobin solutions (660g) of lower floor.
(2), gained sheep hemoglobin solutions is placed closed container, bleed with vavuum pump then, when vacuum reaches 0.087 MPa, be heated to 64 ℃ again, bled 4 minutes, obtain the sheep hemoglobin solutions of without smelling of mutton.
(3), step (2) gained sheep hemoglobin solutions is diluted to concentration is 17% to water, regulating pH is 7.0, (papain and neutral proteinase are in 1: 1 ratio of enzyme work to add compound protease; The source is with embodiment 1) 1.3g, the reaction 5.5h that under 43 ℃, pH7.0 condition, is hydrolyzed is then 85 ℃ of insulations 3 minutes down; Be cooled to 20 ℃; At the centrifugal 8min of 6000g, collecting precipitation obtains sediment 31g.
(4), with the sediment of 40g distilled water washing step (3) gained, centrifugal 8 minutes then at 5000g, taking precipitate; Repeat above-mentioned washing precipitate operation 2 times,, get heme peptide 15g at last with sediment freeze drying (FD-1PF, development in science and technology Co., Ltd is helped in moral sky, Beijing) 15 hours.
Adopt the content of iron in the potassium rhodanide colorimetric method for determining heme peptide, the content of iron is 7.1% in the heme peptide as a result.
The preparation of embodiment 3 sheep heme peptides
Carry out according to following steps:
(1), in the 2000g sheep blood, adding concentration is 20% citric acid three sodium solution 37g; Stir, then at 5000g centrifugal (TGL-16A centrifuge, the ordinary instrument and meter in Changsha Co., Ltd) 6min; Separate and obtain two layers of solution; Lower floor is sheep hemoglobin solutions (1340g), and the upper strata is sheep blood plasma protein solution (660g), collects the sheep hemoglobin solutions of lower floor.
(2), the sheep hemoglobin solutions is placed closed container, bleed with vavuum pump then, when vacuum reaches 0.084 MPa, be heated to 65 ℃ again, bled 5 minutes, obtain the sheep hemoglobin solutions of without smelling of mutton.
(3), the sheep hemoglobin solutions of step (2) gained is diluted to concentration is 18% to water, regulating pH is 6.9, adds compound protease (papain and neutral proteinase are in 1: 1 ratio of enzyme work) 3.0g; At 43 ℃, pH is the reaction 6h that is hydrolyzed under 6.9 conditions; 90 ℃ of insulations 3 minutes, be cooled to 20 ℃, then at the centrifugal 6min of 6000g; Collecting precipitation obtains sediment 60g;
(4), with the sediment of 80g distilled water washing step (3) gained, at 5000g centrifugal 8 minutes, taking precipitate; Repeat above-mentioned washing precipitate operation 2 times,, get heme peptide 31g sediment freeze drying (FD-1PF, development in science and technology Co., Ltd is helped in moral sky, Beijing) 10 hours.
Adopt the content of iron in the potassium rhodanide colorimetric method for determining heme peptide, the content of iron is 7.3% in the heme peptide as a result.
Claims (4)
1. sheep heme peptide is characterized in that according to following method preparation:
(1) in sheep blood, adds trisodium citrate according to 0.3～0.5% percentage by weight, stir, then at the centrifugal 6～10min of 4000～6000g; Be separated into two layers of solution; The upper strata is the sheep blood plasma protein solution, and lower floor is the sheep hemoglobin solutions, collects the sheep hemoglobin solutions of lower floor; Described sheep blood is fresh sheep blood;
(2) gained sheep hemoglobin solutions is placed closed container, bleed with vavuum pump then, when vacuum reaches 0.07～0.09 MPa, be heated to 60～70 ℃ again and bled 3～5 minutes, obtain the sheep hemoglobin solutions of without smelling of mutton;
(3) water is 15～20% with the concentration adjustment of the sheep hemoglobin solutions of step (2) gained; Regulating its pH is 6.5～7.5; According to mass ratio between compound protease and the sheep hemoglobin is that 1: 50～100 ratio adds compound protease in the sheep hemoglobin solutions, is the reaction 4～7h that is hydrolyzed under 6.5～7.5 conditions at 40～45 ℃, pH then, again at 85～90 ℃ of insulation 2～4min enzymes that go out down; Then be cooled to 20 ℃; At the centrifugal 5～10min of 5000～6000g, collecting precipitation gets sediment; Described compound protease is made up of papain and neutral proteinase, and the ratio of living of the enzyme between them is 1: 1;
(4) with the sediment of distilled water washing step (3) gained of 1～2 times of weight, again at the centrifugal 5min～10min of 4000g～5000g, taking precipitate; Press aforesaid operations repeated washing sediment 2～3 times; Freeze drying gets the sheep heme peptide.
2. according to the described sheep heme peptide of claim 1, it is characterized in that hydrochloric acid or the sodium hydroxide solution adjusting of the pH value described in its step (3) with 1 molar concentration.
3. the preparation method of the described sheep heme peptide of claim 1 comprises the steps:
(1) in sheep blood, adds trisodium citrate according to 0.3～0.5% percentage by weight, stir, then at the centrifugal 6～10min of 4000～6000g; Be separated into two layers of solution; The upper strata is the sheep blood plasma protein solution, and lower floor is the sheep hemoglobin solutions, collects the sheep hemoglobin solutions of lower floor; Described sheep blood is fresh sheep blood;
(2) gained sheep hemoglobin solutions is placed closed container, bleed with vavuum pump then, when vacuum reaches 0.07～0.09 MPa, be heated to 60～70 ℃ again and bled 3～5 minutes, obtain the sheep hemoglobin solutions of without smelling of mutton;
(3) the sheep hemoglobin solutions of step (2) gained is diluted to concentration is 15～20% to water; Regulating pH is 6.5～7.5; According to mass ratio between compound protease and the sheep hemoglobin is that 1: 50～100 ratio adds compound protease in the sheep hemoglobin solutions, and the reaction 4～7h that under 40～45 ℃, pH 6.5～7.5 conditions, is hydrolyzed then is again at 85～90 ℃ of insulation 2～4min enzymes that go out down; Then be cooled to 20 ℃; At the centrifugal 5～10min of 5000～6000g, collecting precipitation gets sediment; Described compound protease is made up of papain and neutral proteinase, and the ratio of living of the enzyme between them is 1: 1;
(4) with the sediment of distilled water washing step (3) gained of 1～2 times of weight, again at the centrifugal 5min～10min of 4000g～5000g, taking precipitate; Press aforesaid operations repeated washing sediment 2～3 times; Freeze drying gets the sheep heme peptide.
4. according to the described preparation method of claim 3, it is characterized in that hydrochloric acid or the sodium hydroxide solution adjusting of the pH value described in its step (3) with 1 molar concentration.
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| CN104003769B (en) * | 2014-06-05 | 2016-05-04 | 湖北大学 | A kind of preparation method of duck hemepeptide and duck hemepeptide are in the application of plantation duck blood tea |
| CN108651995A (en) * | 2017-04-01 | 2018-10-16 | 新疆喀什昆仑翠翎鸽业有限责任公司 | One kind is rich in active peptides dove Blood piece agent and preparation method thereof of enriching blood |
| CN108753895A (en) * | 2018-07-02 | 2018-11-06 | 华中农业大学 | A kind of small molecule heme peptide and preparation method thereof |
| CN112481342A (en) * | 2020-12-11 | 2021-03-12 | 湖北省农业科学院农产品加工与核农技术研究所 | Method for synchronously preparing poultry heme and antioxidant blood peptide |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1279061A (en) * | 1999-07-06 | 2001-01-10 | 张大文 | Process for preparing cosmetic lotion containing sheep placenta extract and its deodouring reactor |
| CN101085777A (en) * | 2007-07-10 | 2007-12-12 | 青岛波尔旺肉业股份有限公司 | Method for producing sheep blood hemachrome |
| CN101107943A (en) * | 2007-08-20 | 2008-01-23 | 澳芝妙乳业（北京）有限公司 | Physics the goat milk smell doffing method |
| CN101366442A (en) * | 2008-10-08 | 2009-02-18 | 青岛波尔旺肉业股份有限公司 | Sheep hemoglobin blood pressure reduction peptide and preparation method |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1279061A (en) * | 1999-07-06 | 2001-01-10 | 张大文 | Process for preparing cosmetic lotion containing sheep placenta extract and its deodouring reactor |
| CN101085777A (en) * | 2007-07-10 | 2007-12-12 | 青岛波尔旺肉业股份有限公司 | Method for producing sheep blood hemachrome |
| CN101107943A (en) * | 2007-08-20 | 2008-01-23 | 澳芝妙乳业（北京）有限公司 | Physics the goat milk smell doffing method |
| CN101366442A (en) * | 2008-10-08 | 2009-02-18 | 青岛波尔旺肉业股份有限公司 | Sheep hemoglobin blood pressure reduction peptide and preparation method |
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