CN107557423A - A kind of preparation method of camel hemalbumin polypeptide - Google Patents
A kind of preparation method of camel hemalbumin polypeptide Download PDFInfo
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- CN107557423A CN107557423A CN201711025726.0A CN201711025726A CN107557423A CN 107557423 A CN107557423 A CN 107557423A CN 201711025726 A CN201711025726 A CN 201711025726A CN 107557423 A CN107557423 A CN 107557423A
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- hemalbumin
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- enzymolysis liquid
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Abstract
It is an object of the invention to provide a kind of preparation method of camel hemalbumin polypeptide, to make up the deficiency of camel hemalbumin polypeptide production technology, improves the value of camel hemalbumin polypeptide.Gained camel hemalbumin polypeptide can be used for the raw material of food, health food, special medicine purposes food and medicine.The present invention is during hunchbacked hemalbumin polypeptide is prepared without any material of addition.There is no phase transformation, whole process is carried out at less than 60 DEG C, is effectively protected the quality and effect of hunchbacked hemalbumin polypeptide.The method of the present invention effectively removes small-molecule substance and amino acid in hunchbacked blood, make the content of activated protein polypeptide in its unit mass increase, and ensure the bioactivity of camel hemalbumin.
Description
Technical field：
The invention belongs to polypeptide preparing technical field, and in particular to a kind of preparation method of camel hemalbumin polypeptide.
Background technology
Polypeptide is the intermediate product of protein hydrolysis, simpler than protein structure, and molecular weight is small, generally by three or three
Above amino acid molecular dehydrating condensation forms.Polypeptide is the bioactive substance for being related to various cell functions in organism, almost
All cells all by polypeptides for modulating, have regulation body physiological function and the double effects of nutrition are provided for body.China is generation
One of main place of production of camel in boundary, the whole nation there are about peak of camel 29.5 ten thousand or so, be mainly distributed on Inner Mongol, Xinjiang, Qinghai, sweet
On the arid desert grassland that about 1,100,000 square kilometres of the provinces and regions such as respectful, Ningxia.In camel industrial chain, hunchbacked milk, camel's hair and hunchbacked meat are
Its major product, and slaughtering hunchbacked blood caused by camel, due to reasons such as mouthfeels, in addition to the peasants and herdsmen's use of small part locality,
The overwhelming majority goes out of use, and causes the huge wasting of resources and environmental pollution.
Main active is protein in camel blood, and content is about 15~20%, is huge animal protein storehouse.Camel
With its special function, superpower tolerance and indomitable vitality, run quickly among the desert and Gobi of life disappearance, it is warm
Referred to as " ship of the desert ".Special environment makes its blood compared with the blood of other animals, has special physiology, nutrition work(
Energy.Modern study shows：Albumin in camel blood is lived not only apparently higher than other animals with abnormal high physiology
Property.In the total protein of camel serum, albumin content is up to 73.20%, hence it is evident that is moved higher than mammalitys such as sheep, ox, horses
The albumin of blood in thing.Therefore, using hunchbacked blood as raw material, hunchbacked hemalbumin polypeptide is prepared using modern separation technology separation, and then
The health-oriented products as raw material are researched and developed, this precious resources of desert area is made full use of, has great importance.
The camel hemalbumin polypeptide for the high-purity being prepared is milky, and does not have peculiar smell.And derive from camel blood
The polypeptide crude product of liquid has obvious fishy smell, and is in kermesinus, has had a strong impact on its extensive use and market value.
The content of the invention
It is an object of the invention to provide a kind of preparation method of camel hemalbumin polypeptide, to make up the life of camel hemalbumin polypeptide
The deficiency of production. art, improve the value of camel hemalbumin polypeptide.Gained camel hemalbumin polypeptide can be used for food, health care
The raw material of food, special medicine purposes food and medicine.
The preparation method of the camel hemalbumin polypeptide of the present invention, comprises the following steps：
1) camel blood is added in 3~6 times of water, the Mixed enzyme for animal proteolysis that addition concentration is 0.3%~1% enters
Row processing, hydrolysis temperature are 55~60 DEG C, and enzymolysis time is 1~3h；
2) after enzymolysis terminates, temperature is risen to 90 DEG C rapidly and inactivated, is then cooled, centrifuged, obtain enzymolysis liquid；
3) after enzymolysis liquid being removed into impurity by ceramic membrane, by ultrafiltration membrane system, enzymolysis liquid I is obtained, enzymolysis liquid I is led to
Nanofiltration membrane system is crossed, obtains enzymolysis liquid II；
4) enzymolysis liquid II is dried, sterilizes, packs and obtain camel hemalbumin polypeptide products.
Above-mentioned steps 1) described in Mixed enzyme for animal proteolysis include pepsin, trypsase, serine protease
With in thiol protease it is any two or more.
Above-mentioned steps 2) in the rotating speed of centrifugation be 16000rpm, centrifuge 10min or more.
Above-mentioned steps 3) described in milipore filter molecular weight be 5000~3000,15~30bar of pressure, temperature be less than 30 DEG C；
Above-mentioned steps 3) described in NF membrane molecular weight be 1000~180,5~10bar of pressure, temperature be less than 30 DEG C；
Above-mentioned steps 4) described in drying be freeze-drying, spray drying, vacuum drying, one kind in forced air drying.
The optimal conditions of the spray drying is：EAT：100~145 DEG C, leaving air temp：80~110 DEG C, feed liquid
Density 0.9~1.1.
The optimal conditions of the freeze-drying is：- 60 DEG C~-80 DEG C of cryogenic temperature, solid content 60%~80%.
Described sterilization method includes a kind of or several in Co-60 radiation sterilization, ultraviolet irradiation sterilizing, electron beam sterilization
Kind, sterilization time is 3~6h.
The present invention is during hunchbacked hemalbumin polypeptide is prepared without any material of addition.There is no phase transformation, whole process is low
Carried out at 60 DEG C, be effectively protected the quality and effect of hunchbacked hemalbumin polypeptide.The method of the present invention effectively removes hunchbacked blood
In small-molecule substance and amino acid, make the content of activated protein polypeptide in its unit mass increase, ensure camel hemalbumin
Bioactivity.
Embodiment
In the prior art, the fishy smell and color and luster of hunchbacked hemalbumin polypeptide products are removed, turns into and hinders its high-valued key
Technical problem.At present, the method that common deodorant is decolourized has a hydrogen peroxide method, activated carbon method and Flavonoids by Macroporous Adsorption Resin etc., its
Middle hydrogen peroxide method can cause the composition in polypeptide to aoxidize；Activated carbon method can adsorb substantial amounts of polypeptide, cause
Active material is lost in, and yield declines；Flavonoids by Macroporous Adsorption Resin can not only cause active material that dead absorption occurs, it is difficult to elutes, and
And the production that macroporous absorbent resin is used for food, health food has significant limitation, cost height.Therefore, development is a kind of new
The production technology that the camel hemalbumin polypeptide deodorant of type is decolourized, has great importance.
It is embodiments of the invention below, the above of the present invention is described in further detail.But not should
This scope for being interpreted as aforementioned body of the present invention is only limitted to following embodiment.It should be pointed out that for the common of the art
For technical staff, do not depart from the present invention core technology on the premise of, improvements and modifications can also be made, these improve and
Retouching should also belong to the scope of patent protection of the present invention.It is any in the implication and scope suitable with claims of the present invention
Change, be all considered as being included within the scope of the claims.
Embodiment 1：
1) 5L purified waters will be added in the fresh camel blood of 1kg, extraction is transferred to after shearing 30min with high-speed shearing machine
Tank, add pepsin 15g, trypsase 15g, 60 DEG C of Extracting temperature；Stir enzyme digestion reaction 2h；It is brought rapidly up to after 90 DEG C,
0.5h is reacted, is then centrifuged with 16000rpm tube centrifuge, obtains enzymolysis liquid.
2) enzymolysis liquid is pumped into ceramic membrane, the enzymolysis liquid of permeation ceramic membrane passes through the membranous system that molecular weight is 5000, mistake again
Film temperature is controlled below 25 DEG C, flow velocity 50mL/min, pressure 15bar, collects the enzymolysis liquid through membranous system；
3) will transmit through the enzymolysis liquid of membranous system by molecular weight is 350 nanofiltration membrane systems, crosses film temperature control at 30 DEG C, pressure
Power is 25bar.The enzymolysis liquid Direct spraying of membranous system retention is dried, the white powder of obtained milk is consolidated for hunchbacked hemalbumin polypeptide
Body, after aluminium foil bag packing, Co-60 radiation sterilizing equipments sterilizing 3h is placed in, produces hunchbacked hemalbumin polypeptide finished product, yield 10%.
Products obtained therefrom detects through HPLC, molecular weight ranges be 5000~800, its middle-molecular-weihydroxyethyl concentrate on 1000 or so it is more
Peptide accounts for more than 70%.
Hunchbacked hemalbumin polypeptide anti-fatigue active evaluation
1. experimental method
1.1 experimental animal：ICR adult male mices, 18~22g of body weight.
1.2 dosage are grouped and given the test agent gives the time
Experiment sets hunchbacked two dosage groups (100mg/kg, 200mg/kg) of hemalbumin polypeptide, a hunchbacked hemalbumin dosage group
(200mg/kg) and negative control dosage group.Given the test agent is given 30 days, will be every after last gives given the test agent 30min
The sheet lead of 10 body weight of mouse tail root load 5% of group, is placed in swimming with a load attached to the body in the swimming trunk of (30 ± 2 DEG C).Swum with mouse
Swimming to head gos deep into 10s in water and no longer floats end, records mouse swimming time.Remaining every group of 10 mouse takes blood,
Determine blood lactase acid.
2. time interpretation of result
Influence of the 2.1 hunchbacked hemalbumin polypeptides to tired mouse movement endurance, blood lactase acid
In terms of power exhausts the time：Hunchbacked two dosage groups 100mg/kg, 200mg/kg of hemalbumin polypeptide are compared with negative control
With significant difference, power exhausts the time and extends 27% and 35% respectively；Meanwhile hunchbacked two dosage group (100mg/ of hemalbumin polypeptide
Kg, 200mg/kg) compared with (200mg/kg) hunchbacked hemalbumin dosage group power exhausts the time also there are significant difference, and 100mg/kg
Hunchbacked hemalbumin polypeptide dosage group exhaust time lengthening 7% than 200mg/kg camel hemalbumin dosage group power.
In terms of blood lactase acid：Hunchbacked two dosage groups (100mg/kg, 200mg/kg) of hemalbumin polypeptide are compared with negative control
With significant difference, blood lactase acid area under the curve is decreased obviously, and have dropped 12.3% and 18.7% respectively；And 200mg/kg camel blood
Albumen dosage group blood lactase acid area under the curve compares no difference with control.
Thus illustrate that the hunchbacked hemalbumin polypeptide that patent of the present invention obtains has good antifatigue effect, and positive effect is excellent
In hunchbacked hemalbumin.This may make it that the molecular weight of hunchbacked hemalbumin polypeptide is low with this preparation method, and branched-chain amino acid is more, utilizes suction
Receive, bioavilability height is relevant.
Embodiment 2：
1) 40kg camel blood is placed in 500L enzymatic vessel, adds 200L purified waters.Stirring is warming up to 60 DEG C, herein
During, by 1.2kg pepsin, trypsase, thiol protease mixture (1:1:0.5) add 1L and be less than 60 DEG C
Enzymatic vessel is added after fully being dissolved in purified water, stops rising to after temperature rises to 60 DEG C, keeps 60 DEG C of reaction 2h.Enzymolysis terminates
After be brought rapidly up to 90 DEG C of reaction 0.5h, enzymolysis liquid is squeezed into storage tank, outer circulation adds industrial water auxiliary temperature-reducing；Temperature is down to
At less than 30 DEG C, tube centrifuge centrifugation is pumped into, rotating speed is in 12000rpm.
2) above-mentioned enzymolysis liquid is pumped into ceramic membrane, from ceramic membrane pass through enzymolysis liquid then by molecular weight be 5000 film
System, the membranous system of 350 molecular weight is pumped into again through the enzymolysis liquid of 5000 membranous systems, whole process is entered in the case of sealing
OK, temperature control is below 60 DEG C, and pressure is in 20bar or so.
3) enzymolysis liquid by the retention of 350 membranous systems is collected, and is spray-dried and is produced hunchbacked hemalbumin polypeptide powder, yield 10%, milk
White, free from extraneous odour, there is light milk fragrance.It will detect, indices all meet and are better than national solid beverage standard (NY/T
1323-2007)。
Claims (10)
1. a kind of preparation method of camel hemalbumin polypeptide, it is characterised in that described preparation method comprises the following steps：
1) camel blood is added in 3~6 times of water, addition concentration is at 0.3%~1% Mixed enzyme for animal proteolysis
Reason, hydrolysis temperature are 55~60 DEG C, and enzymolysis time is 1~3h；
2) after enzymolysis terminates, temperature is risen to 90 DEG C rapidly and inactivated, is then cooled, centrifuged, obtain enzymolysis liquid；
3) after enzymolysis liquid being removed into impurity by ceramic membrane, by ultrafiltration membrane system, enzymolysis liquid I is obtained, by enzymolysis liquid I by receiving
Filter membrane system, obtain enzymolysis liquid II；
4) enzymolysis liquid II is dried, sterilizes, packs and obtain camel hemalbumin polypeptide products.
2. the method as described in claim 1, it is characterised in that the Mixed enzyme for animal proteolysis of described step 1) includes stomach
In protease, trypsase, serine protease and thiol protease it is any two or more.
3. the method as described in claim 1, it is characterised in that the rotating speed of centrifugation is 16000rpm in described step 2), from
Heart 10min or more.
4. the method as described in claim 1, it is characterised in that milipore filter molecular weight in described step 3) for 5000~
3000。
5. the method as described in claim 1, it is characterised in that NF membrane molecular weight in described step 3) for 1000~
180。
6. the method as described in claim 1, it is characterised in that the drying in described step 4) is freeze-drying, spraying is dry
Any of dry, vacuum drying, forced air drying.
7. method as claimed in claim 6, it is characterised in that the condition being spray-dried described in described step 4) is as follows：
EAT：100~145 DEG C, leaving air temp：80~110 DEG C, feed liquid density 0.9~1.1.
8. the method as described in claim 1, it is characterised in that the sterilization method in described step 4) radiates including Co-60
One or several kinds in sterilizing, ultraviolet irradiation sterilizing, electron beam sterilization.
9. a kind of camel hemalbumin polypeptide, it is characterised in that described polypeptide is prepared with any methods of claim 1-8
's.
10. application of the polypeptide in antifatigue product is prepared described in claim 9.
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Cited By (5)
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| CN105596370A (en) * | 2015-10-13 | 2016-05-25 | 中国科学院兰州化学物理研究所 | Preparation method of camel blood polypeptides |
| CN111607628A (en) * | 2020-06-02 | 2020-09-01 | 中国科学院兰州化学物理研究所 | A kind of glycopeptide for lowering blood sugar of radix Pachyrhizi Erosi and camel |
| CN111820314A (en) * | 2020-08-04 | 2020-10-27 | 中国科学院兰州化学物理研究所 | An antifatigue health product prepared from sanguis Cervi polypeptide and fructus Lycii polysaccharide |
| CN113304244A (en) * | 2021-05-26 | 2021-08-27 | 杭州佰倍优生物科技有限公司 | Preparation method of blood protein polypeptide for preventing and relieving anemia |
| CN113797320A (en) * | 2021-11-01 | 2021-12-17 | 杭州佰倍优生物科技有限公司 | Polypeptide composition for improving anemia symptoms and preparation method thereof |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105596370A (en) * | 2015-10-13 | 2016-05-25 | 中国科学院兰州化学物理研究所 | Preparation method of camel blood polypeptides |
| CN111607628A (en) * | 2020-06-02 | 2020-09-01 | 中国科学院兰州化学物理研究所 | A kind of glycopeptide for lowering blood sugar of radix Pachyrhizi Erosi and camel |
| CN111607628B (en) * | 2020-06-02 | 2022-10-25 | 中国科学院兰州化学物理研究所 | A kind of glycopeptide for lowering blood sugar of radix Pachyrhizi Erosi and camel |
| CN111820314A (en) * | 2020-08-04 | 2020-10-27 | 中国科学院兰州化学物理研究所 | An antifatigue health product prepared from sanguis Cervi polypeptide and fructus Lycii polysaccharide |
| CN113304244A (en) * | 2021-05-26 | 2021-08-27 | 杭州佰倍优生物科技有限公司 | Preparation method of blood protein polypeptide for preventing and relieving anemia |
| CN113797320A (en) * | 2021-11-01 | 2021-12-17 | 杭州佰倍优生物科技有限公司 | Polypeptide composition for improving anemia symptoms and preparation method thereof |
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