CN109824754A - A kind of yak skin polypeptide and preparation method thereof with nourishing blood function - Google Patents
A kind of yak skin polypeptide and preparation method thereof with nourishing blood function Download PDFInfo
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- CN109824754A CN109824754A CN201910206116.3A CN201910206116A CN109824754A CN 109824754 A CN109824754 A CN 109824754A CN 201910206116 A CN201910206116 A CN 201910206116A CN 109824754 A CN109824754 A CN 109824754A
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Abstract
The present invention relates to medicine field more particularly to a kind of yak skin polypeptides and preparation method thereof with nourishing blood function.The yak skin polypeptide molecular weight is made of 17KDa, 34 KDa, 43 KDa, and peptide bond ultraviolet maximum absorption wavelength is at 216 nm, and amide I band infrared spectroscopy is in 1600 ~ 1700 cm‑1Place, amide II band infrared spectroscopy is in 1560 ~ 1500 cm‑1Place, amide III infrared spectroscopy is in 1234 ~ 1237 cm‑1Place.Preparation method includes mill skin, heats, mentions foam, prepares enzymolysis liquid, centrifugal concentrating and separation drying.The present invention is full of nutrition using from pure natural free of contamination yak in Qinghai-tibet skin, as primary raw material, raw material sources are extensive, increases a kind of product with nourishing blood function while improving yak skin economic value added；Its preparation manipulation process is simple, and the splitter of early period is changed to decorticating mill, avoid because raw material thickness it is different caused by waste；There are three the bioactive ingredients of enriching blood of molecular weight section for yak skin polypeptide tool obtained.
Description
Technical field
The present invention relates to medicine field more particularly to a kind of yak skin polypeptides and preparation method thereof with nourishing blood function.
Background technique
Yak is the peculiar ox kind in plateau, it is lived in, and anoxic, high and cold, High aititude, herbage are short, sparse, withered grass phase length
In severe living environment.Glue nutritional ingredient rich in, document report Yak-skin Gelatin made of its fresh hide is processed contain
20.95 % human bodies must the mineral matter elements such as amino acid and calcium, sodium, magnesium, potassium.Previous research work finds different molecular weight section
Yak-skin Gelatin is in physicochemical property --- denaturation temperature, emulsifiability, foaming capacity and foamed stability, antioxygenic property etc. have compared with
Big difference.Therefore, the economic value of yak not only can be improved in exploitation yak skin product, equally has to Community health's public health work
There is positive effect.
Anaemia is in the disease incidence highest of asian population, and since the period is long, progression of the disease is slow, not specific clinical condition
Shape has great hiding, and health and quality of life to people cause to seriously affect, its cognition and fortune to children
Dynamic development has a negative impact, and leads to the fatigue and low productivity of general population.Modern studies have found that Yak-skin Gelatin is to human body
Alimentary anemia plays the role of improvement, and in applicant's previous research, to the research of Yak-skin Gelatin have multinomial authorization and
Accept patent of invention.Wherein, the extracting and developing of Yak-skin Gelatin has obtained authorization patent of invention.For this purpose, in the base of early-stage study
It is the scope of application of further expansion yak skin on plinth, the applicant has studied the nourishing blood function of Yak-skin Gelatin again.
Summary of the invention
The purpose of the present invention is to provide a kind of yak skin polypeptide and preparation method thereof with nourishing blood function, especially by
The splitter of early period is changed to decorticating mill by yak skin dermal sites, avoid because raw material thickness it is different caused by waste, adopted after extraction
The suitable molecular weight ranges of Yak-skin Gelatin, and the yak skin polypeptide with nourishing blood function are obtained with different UF membranes.
To achieve the above object, the technical solution of the present invention is as follows:
A kind of yak skin polypeptide with nourishing blood function, molecular weight are made of 17KDa, 34 KDa, 43 KDa, the ultraviolet maximum of peptide bond
Absorbing wavelength is at 216 nm, and amide I band infrared spectroscopy is in 1600 ~ 1700 cm-1Place, amide II band infrared spectroscopy 1560 ~
1500 cm-1Place, amide III infrared spectroscopy is in 1234 ~ 1237 cm-1Place.
A kind of preparation method of the yak skin polypeptide with nourishing blood function, includes the following steps:
(1) it grinding skin: the yak skin after degreasing of losing hair or feathers is cleaned up, clipper is cut into the square of 10~15cm, epidermis is ground off,
It cleans up；
(2) heat: it is 1~9 that pH, which is added, in the ratio of 1:1~10 in the yak skin for grinding off epidermis, and concentration is the sweet of 0.1 mol/L
In propylhomoserin buffer, 2~30 min are heated, are cooled to room temperature；
(3) it mentions foam: taking pure water, be heated to water boiling, be put into yak Pi Timo, 0.1~0.5 h of time；
(4) prepare enzymolysis liquid: being enzyme: yak skin=1:1~100(w/w according to material ratio), hydrolysis temperature is 20~45 °C, enzymatic hydrolysis
Time is 0.5~24 h, and enzymolysis liquid is heated 2~30 min, is cooled to room temperature；
Optimization, the preparation enzymolysis liquid: being enzyme: yak skin=1:1~50(w/w according to material ratio), hydrolysis temperature is 20~
45 °C, enzymolysis time is 0.5~12 h, and enzymolysis liquid is heated 2~30 min, is cooled to room temperature；
(5) centrifugal concentrating: by the enzymolysis liquid centrifugal force 13500*g of step (4), being centrifuged 2~30 min, be collected into supernatant, dense
It is reduced to certain proportion；
(6) it separating drying: taking yak skin concentrate, Ultra filtration membrane, membrane aperture is 0.02~2.0 μm, it is dry, obtain molecule
Measure the yak skin polypeptide being made of 17Ka, 34 KDa, 43 KDa.
Further, yak skin as described in step (1) is the skin of yak in Qinghai-tibet.
Further, mill skin as described in step (1) is carried out in decorticating mill.
Further, step (4) prepare enzymolysis liquid described in enzyme be pepsin, papain, alkali protease,
Compound protease it is one or more.
Further, heating described in above-mentioned preparation method is microwave heating.
Further, drying described in step (6) is freeze-drying.
Yak skin polypeptide made from the above method has nourishing blood function.
Compared with prior art, the beneficial effects of the present invention are: the present invention is to come from pure natural free of contamination Qinghai-Tibet Platean
Yak skin is primary raw material, and raw material sources are extensive, full of nutrition, increases one kind while improving yak skin economic value added
Product with nourishing blood function；Its preparation manipulation process is simple, and the splitter of early period is changed to decorticating mill, is avoided because of raw material thickness
It is wasted caused by different；There are three the bioactive ingredients of enriching blood of molecular weight section for yak skin polypeptide tool obtained.
Detailed description of the invention
Fig. 1 is that 14 d spleen HE coloration results (HE 400 ×) are administered to anemia mice in Yak-skin Gelatin polypeptide of the present invention
In figure: A is blank control group；B is model group；C, D, E are Yak-skin Gelatin polypeptide group；F is donkey-hide gelatin group.
Specific embodiment
Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out clear, complete
Site preparation description, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.It is based on
Embodiment in the present invention, it is obtained by those of ordinary skill in the art without making creative efforts every other
Embodiment shall fall within the protection scope of the present invention.
Embodiment 1
A kind of yak skin polypeptide with nourishing blood function, molecular weight are made of 17KDa, 34 KDa, 43 KDa, the ultraviolet maximum of peptide bond
Absorbing wavelength is at 216 nm, and amide I band infrared spectroscopy is in 1600 ~ 1700 cm-1Place, amide II band infrared spectroscopy 1560 ~
1500 cm-1Place, amide III infrared spectroscopy is in 1234 ~ 1237 cm-1Place.
A kind of preparation method of the yak skin polypeptide with nourishing blood function, includes the following steps:
(1) it grinds skin: the yak in Qinghai-tibet skin after degreasing of losing hair or feathers being cleaned up, clipper is cut into the square of 10cm, sets mill
Epidermis is ground off in skin machine, is cleaned up；
(2) heat: it is 1 that pH, which is added, in the ratio of 1:1 in the yak skin for grinding off epidermis, and the glycine that concentration is 0.1 mol/L is slow
In fliud flushing, 2 min of microwave heating is cooled to room temperature；
(3) it mentions foam: taking pure water, be heated to water boiling, be put into yak Pi Timo, time 0.1h；
(4) enzymolysis liquid is prepared: according to material ratio pepsin: yak skin=1:1(w/w), hydrolysis temperature is 20 °C, enzymolysis time
2 min of enzymolysis liquid microwave heating is cooled to room temperature for 0.5 h；
(5) centrifugal concentrating: by the enzymolysis liquid centrifugal force 13500*g of step (4), 2 min is centrifuged, supernatant is collected into, is concentrated into
Certain proportion；
(6) it separates drying: taking yak skin concentrate, Ultra filtration membrane, membrane aperture is 0.02 μm, and freeze-drying obtains molecular weight
The yak skin polypeptide being made of 17Ka, 34 KDa, 43 KDa.
Yak skin polypeptide made from the above method has nourishing blood function.
Embodiment 2
A kind of yak skin polypeptide with nourishing blood function, molecular weight are made of 17KDa, 34 KDa, 43 KDa, the ultraviolet maximum of peptide bond
Absorbing wavelength is at 216 nm, and amide I band infrared spectroscopy is in 1600 ~ 1700 cm-1Place, amide II band infrared spectroscopy 1560 ~
1500 cm-1Place, amide III infrared spectroscopy is in 1234 ~ 1237 cm-1Place.
A kind of preparation method of the yak skin polypeptide with nourishing blood function, includes the following steps:
(1) it grinds skin: the yak in Qinghai-tibet skin after degreasing of losing hair or feathers being cleaned up, clipper is cut into the square of 15cm, sets mill
Epidermis is ground off in skin machine, is cleaned up；
(2) heat: it is 9 that pH, which is added, in 1~10 ratio in the yak skin for grinding off epidermis, and concentration is the glycine of 0.1 mol/L
In buffer, 30 min of microwave heating is cooled to room temperature；
(3) it mentions foam: taking pure water, be heated to water boiling, be put into yak Pi Timo, 0.5 h of time；
(4) enzymolysis liquid is prepared: according to material ratio pepsin+papain+alkali protease+compound protease: yak skin=
1:100(w/w), hydrolysis temperature is 45 °C, and enzymolysis time is that 30 min of enzymolysis liquid microwave heating is cooled to room temperature by 24 h；
(5) centrifugal concentrating: by the enzymolysis liquid centrifugal force 13500*g of step (4), 30 min is centrifuged, supernatant is collected into, is concentrated into
Certain proportion；
(6) it separates drying: taking yak skin concentrate, Ultra filtration membrane, membrane aperture is 2.0 μm, and freeze-drying obtains molecular weight
The yak skin polypeptide being made of 17Ka, 34 KDa, 43 KDa.
Yak skin polypeptide made from the above method has nourishing blood function.
Embodiment 3
A kind of yak skin polypeptide with nourishing blood function, molecular weight are made of 17KDa, 34 KDa, 43 KDa, the ultraviolet maximum of peptide bond
Absorbing wavelength is at 216 nm, and amide I band infrared spectroscopy is in 1600 ~ 1700 cm-1Place, amide II band infrared spectroscopy 1560 ~
1500 cm-1Place, amide III infrared spectroscopy is in 1234 ~ 1237 cm-1Place.
A kind of preparation method of the yak skin polypeptide with nourishing blood function, includes the following steps:
(1) it grinds skin: the yak in Qinghai-tibet skin after degreasing of losing hair or feathers being cleaned up, clipper is cut into the square of 13cm, sets mill
Epidermis is ground off in skin machine, is cleaned up；
(2) heat: it is 4 that pH, which is added, in the ratio of 1:5 in the yak skin for grinding off epidermis, and the glycine that concentration is 0.1 mol/L is slow
In fliud flushing, 15 min of microwave heating is cooled to room temperature；
(3) it mentions foam: taking pure water, be heated to water boiling, be put into yak Pi Timo, 0.3 h of time；
(4) enzymolysis liquid is prepared: according to material ratio pepsin+papain: yak skin=1:50(w/w), hydrolysis temperature is
35 °C, enzymolysis time is that 17 min of enzymolysis liquid microwave heating is cooled to room temperature by 12 h；
(5) centrifugal concentrating: by the enzymolysis liquid centrifugal force 13500*g of step (4), 18 min is centrifuged, supernatant is collected into, is concentrated into
Certain proportion；
(6) it separates drying: taking yak skin concentrate, Ultra filtration membrane, membrane aperture is 0.06 μm, and freeze-drying obtains molecular weight
The yak skin polypeptide being made of 17Ka, 34 KDa, 43 KDa.
Yak skin polypeptide made from the above method has nourishing blood function.
Embodiment 4
A kind of yak skin polypeptide with nourishing blood function, molecular weight are made of 17KDa, 34 KDa, 43 KDa, the ultraviolet maximum of peptide bond
Absorbing wavelength is at 216 nm, and amide I band infrared spectroscopy is in 1600 ~ 1700 cm-1Place, amide II band infrared spectroscopy 1560 ~
1500 cm-1Place, amide III infrared spectroscopy is in 1234 ~ 1237 cm-1Place.
A kind of preparation method of the yak skin polypeptide with nourishing blood function, includes the following steps:
(1) it grinds skin: the yak in Qinghai-tibet skin after degreasing of losing hair or feathers being cleaned up, clipper is cut into the square of 11.5cm, sets
Epidermis is ground off in decorticating mill, is cleaned up；
(2) heat: it is 7.5 that pH, which is added, in the ratio of 1:6.8 in the yak skin for grinding off epidermis, and concentration is the sweet ammonia of 0.1 mol/L
In acid buffer, 8 min of microwave heating is cooled to room temperature；
(3) it mentions foam: taking pure water, be heated to water boiling, be put into yak Pi Timo, 0.4 h of time；
(4) enzymolysis liquid is prepared: according to material ratio pepsin+papain+alkali protease: yak skin=1:70(w/w),
Hydrolysis temperature is 42 °C, and enzymolysis time is that 20 min of enzymolysis liquid microwave heating is cooled to room temperature by 18 h；
(5) centrifugal concentrating: by the enzymolysis liquid centrifugal force 13500*g of step (4), 20 min is centrifuged, supernatant is collected into, is concentrated into
Certain proportion；
(6) it separates drying: taking yak skin concentrate, Ultra filtration membrane, membrane aperture is 0.09 μm, and freeze-drying obtains molecular weight
The yak skin polypeptide being made of 17Ka, 34 KDa, 43 KDa.
Yak skin polypeptide made from the above method has nourishing blood function.
Embodiment 5
Yak skin polypeptide has nourishing blood function verification test
With healthy Kunming small white mouse, (half male and half female, 4 week old, weight is at 18 ~ 22 grams, by Gansu Chinese of Traditional Chinese Medicine SPF grades of experimental animal
It is experimental animal that center, which provides [quality certification number is SCXK (sweet)], for Anemia model animal, by product of the present invention to experimental group
Mouse continuous gavage is administered 14 days, and sets blank control group, positive controls, 14 days observation each group mouse of administration group successive administration
General signs detect peripheral hemogram, calculate spleen and thymus gland coefficient, as a result as follows: (being detailed in attached drawing 1):
(1) morphosis of the Yak-skin Gelatin polypeptide to anemia mice spleen
The results show that model group red pulp white pulp boundary is unclear, lymphocyte number and acini lienalis are reduced, yak skin polypeptide administration group
Spleen lymphocyte increases, and the boundary of white pulp and red pulp becomes clear.
(2) influence of the Yak-skin Gelatin polypeptide to anemia mice organ coefficient
Yak-skin Gelatin polypeptide group can be such that Spleen coefficient significantly reduces, and so that thymus gland coefficient is extended significant increase with administration time and (be shown in Table
1).
Influence (n=10) of the 1 Yak-skin Gelatin polypeptide of table to anemia mice organ coefficient
Note: compared to the blank group* P< 0.05,** P<0.01；Compared with model group# P< 0.05,## P<0.01
(3) influence of the Yak-skin Gelatin polypeptide to the red system's parameter of anemia mice blood picture
The administration of Yak-skin Gelatin polypeptide, RBC, HGB, HCT, MCH and MCHC are significantly increased.(see Table 2).
Influence (n=10, x ± s) of the 2 Yak-skin Gelatin polypeptide of table to the red system's parameter of anemia mice blood picture
Note: P < 0.01 * P < 0.05 compared to the blank group, * *；#P < 0.05 compared with model group, ##P < 0.01
Erythrocyte number: model group and blank group have significant difference, and Yak-skin Gelatin polypeptide can dramatically increase erythrocyte number,
It is consistent with positive group donkey-hide gelatin group.
Hemoglobin content: model group and blank group have significant difference, and Yak-skin Gelatin polypeptide can dramatically increase blood red egg
White quantity, it is consistent with positive group donkey-hide gelatin group.
Hematocrit: model group and blank group have significant difference, and Yak-skin Gelatin polypeptide can dramatically increase red blood cell
Hematocrit, it is consistent with positive group donkey-hide gelatin group.
Average hemoglobin amount: model group and blank group have significant difference, and Yak-skin Gelatin polypeptide can dramatically increase flat
Equal hemoglobin content, it is consistent with positive group donkey-hide gelatin group.
Mean hemoglobin concentration: model group and blank group have significant difference, and Yak-skin Gelatin polypeptide can dramatically increase
Mean hemoglobin concentration, it is consistent with positive group donkey-hide gelatin group.
In conclusion product of the present invention can significantly improve the morphosis of anemia mice spleen, drench anemia mice spleen
The boundary of bar cytosis, white pulp and red pulp becomes clear, and administration group can be such that Spleen coefficient significantly reduces, and make thymus gland coefficient with administration
Time, which extends, significantly to be increased, and is increased erythrocyte number, is improved content of hemoglobin, and being indicated above products obtained therefrom of the present invention has benefit
Blood effect.
It although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
A variety of variations, modification, replacement can be carried out to these embodiments without departing from the principles and spirit of the present invention by understanding
And modification, the scope of the present invention is defined by the appended.
Claims (9)
1. a kind of yak skin polypeptide with nourishing blood function, which is characterized in that the yak skin polypeptide molecular weight is by 17KDa, 34
KDa, 43 KDa composition, peptide bond ultraviolet maximum absorption wavelength is at 216 nm, and amide I band infrared spectroscopy is in 1600 ~ 1700 cm-1
Place, amide II band infrared spectroscopy is in 1560 ~ 1500 cm-1Place, amide III infrared spectroscopy is in 1234 ~ 1237 cm-1Place.
2. a kind of preparation method of the yak skin polypeptide with nourishing blood function of claim 1, which is characterized in that including walking as follows
It is rapid:
Mill skin: the yak skin after degreasing of losing hair or feathers is cleaned up, clipper is cut into the square of 10~15cm, grinds off epidermis, clearly
Wash clean；
Heating: it is 1~9 that pH, which is added, in the ratio of 1:1~10 in the yak skin for grinding off epidermis, and concentration is the sweet ammonia of 0.1 mol/L
In acid buffer, 2~30 min are heated, are cooled to room temperature；
It mentions foam: taking pure water, be heated to water boiling, be put into yak Pi Timo, 0.1~0.5 h of time；
Prepare enzymolysis liquid: being enzyme: yak skin=1:1~100(w/w according to material ratio), hydrolysis temperature is 20~45 °C, when enzymatic hydrolysis
Between be 0.5~24 h, by enzymolysis liquid heat 2~30 min, be cooled to room temperature；
Centrifugal concentrating: by the enzymolysis liquid centrifugal force 13500*g of step (4), 2~30 min is centrifuged, supernatant is collected into, is concentrated into
Certain proportion；
Separation drying: taking yak skin concentrate, Ultra filtration membrane, and membrane aperture is 0.02~2.0 μm, dry, obtain molecular weight by
The yak skin polypeptide that 17Ka, 34 KDa, 43 KDa are formed.
3. a kind of preparation method of yak skin polypeptide with nourishing blood function according to claim 2, which is characterized in that step
Suddenly yak skin described in (1) is the skin of yak in Qinghai-tibet.
4. a kind of preparation method of yak skin polypeptide with nourishing blood function according to claim 2, which is characterized in that step
Suddenly mill skin described in (1) is carried out in decorticating mill.
5. a kind of preparation method of yak skin polypeptide with nourishing blood function according to claim 2, which is characterized in that step
Suddenly preparation enzymolysis liquid described in (4) is enzyme: yak skin=1:1~50(w/w according to material ratio), hydrolysis temperature is 20~45 °C,
Enzymolysis time is 0.5~12 h, and enzymolysis liquid is heated 2~30 min, is cooled to room temperature.
6. a kind of preparation method of yak skin polypeptide with nourishing blood function according to claim 2, which is characterized in that step
Suddenly (4) prepare enzymolysis liquid described in enzyme be pepsin, papain, alkali protease, compound protease one kind or
It is a variety of.
7. a kind of preparation method of yak skin polypeptide with nourishing blood function according to claim 2, which is characterized in that on
Stating heating described in preparation method is microwave heating.
8. a kind of preparation method of yak skin polypeptide with nourishing blood function according to claim 2, which is characterized in that step
Suddenly drying described in (6) is freeze-drying.
9. special according to a kind of described in any item preparation methods of the yak skin polypeptide with nourishing blood function of claim 2-8
Sign is that obtained yak skin polypeptide has nourishing blood function.
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| CN110229864A (en) * | 2019-06-21 | 2019-09-13 | 中国科学院西北高原生物研究所 | A kind of animal glue preparation method promoting platelet activation |
| CN110229862A (en) * | 2019-06-21 | 2019-09-13 | 中国科学院西北高原生物研究所 | A kind of Yak-skin Gelatin preparation method promoting platelet activation |
| CN110229861A (en) * | 2019-06-21 | 2019-09-13 | 中国科学院西北高原生物研究所 | A kind of animal glue preparation method improving alopecia |
| CN112335882A (en) * | 2020-12-03 | 2021-02-09 | 中国科学院西北高原生物研究所 | Yak skin glue compound product with blood replenishing and bleeding stopping effects and preparation method thereof |
| CN112426483A (en) * | 2020-12-03 | 2021-03-02 | 中国科学院西北高原生物研究所 | Compound yak hide product with blood replenishing effect and preparation method thereof |
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| CN110229864A (en) * | 2019-06-21 | 2019-09-13 | 中国科学院西北高原生物研究所 | A kind of animal glue preparation method promoting platelet activation |
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| CN110229861A (en) * | 2019-06-21 | 2019-09-13 | 中国科学院西北高原生物研究所 | A kind of animal glue preparation method improving alopecia |
| CN112335882A (en) * | 2020-12-03 | 2021-02-09 | 中国科学院西北高原生物研究所 | Yak skin glue compound product with blood replenishing and bleeding stopping effects and preparation method thereof |
| CN112426483A (en) * | 2020-12-03 | 2021-03-02 | 中国科学院西北高原生物研究所 | Compound yak hide product with blood replenishing effect and preparation method thereof |
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