CN103820517B - The preparation method of the yak skin micromolecular collagen of regulation and control human body energy metabolism - Google Patents
The preparation method of the yak skin micromolecular collagen of regulation and control human body energy metabolism Download PDFInfo
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Abstract
The present invention relates to a kind of regulate and control the preparation method of yak skin micromolecular collagen of human body energy metabolism, the method comprises the following steps: (1) by fresh or through infiltrating, softening after the Qinghai-Tibet Platean peculiar ox kind yak wild of drying? the yak skin of yak cleans up, then add alkali to soak, obtain the yak skin after losing hair or feathers; (2) after the yak skin after depilation soaks dealkalize, clear water rinses, and obtains the yak skin of dealkalize depilation; (3) the yak skin of dealkalize depilation cuts into fritter, and then add water skin of scalding, and obtains the yak skin after degreasing; (4) the yak skin water after degreasing carries out extracting rear filtration, discards residue, obtains filtrate; (5) be separated after filtrate water dilution, obtain the former protein solution of the micromolecular Yak-skin Gelatin of 4Kda; (6), by concentrated, dry for micromolecular for 4Kda Yak-skin Gelatin former protein solution warp, obtain yak skin micromolecular collagen.The inventive method is simple, Be very effective, and the former molecular weight of albumen distribution of obtained Yak-skin Gelatin is concentrated, and has the function of regulation and control human body energy metabolism.
Description
Technical field
The present invention relates to a kind of preparation method of collagen protein, particularly relate to the preparation method of the yak skin micromolecular collagen of regulation and control human body energy metabolism.
Background technology
The sugar of energy derive needed for body in food, fat and protein, these nutritive ingredients are transformed into absorbable small molecules nutritive substance through digestion and are absorbed.Human body nearly 16% is protein, and has the collagen protein of 35 ~ 45% in protein, and there are skin, muscle, bone, tooth etc. in the main position existed, and collagen protein is the important composition composition in skeleton, especially cartilaginous tissue.
Serum lactic dehydrogenase (LDH) is a kind of glycolytic ferment, be present in all histiocytic kytoplasms of body, reduce and oxidizing reaction between catalysis pyruvic acid and lactic acid, pyruvic acid is reduced to lactic acid when glycolysis-speed exceedes aerobic metabolism speed, is the important enzyme of involved in sugar anaerobic glycolysis and glyconeogenesis.
In the cytoplasm that creatine kinase (CK) is present in the tissue such as heart, muscle and brain and plastosome; be that one operates with intracellular energy, Muscle contraction, ATP regenerate the important kinases having direct relation, its phosphoryl that turns reversibly between catalysis creatine and ATP reacts.
Triphosaden (ATP) directly supplies the material of energy, PCr is main margin of energy material, under physiological conditions, ATP can be shifted from mitochondrial inner membrane by mitochondrial creatine kinase and form PCr, and PCr is needing to transfer ATP to for utilization through the catalysis of kytoplasm CK isozyme again in position.
Yak is one of ancient and original ox kind of China, is the ox kind of procreation of uniquely surviving under Qinghai-Tibet ecotope, is Qinghai-Tibet distinctive prepotent sire, has unique ecological characteristics and economic worth.Through the Natural Selection and Evolution of 1,100, define the heritability of its uniqueness and the adaptive faculty to severe cold region severe environment, there is extremely strong anti-adversity ability, resistance against diseases, tolerance and adaptability, the high and cold severe ecotope that the immalleable High aititude of general domestic animal, high anoxic etc. are harsh can be adapted to.Biologically active substance containing many antifatigues, anti-hypoxia in yak body.The micromolecular collagen of pure natural is extracted from yak skin, be converted to after being absorbed by body and participate in enzyme needed in human body energy metabolic process or participate in intermediate product anabolic process, energy can be provided effectively in time when body requirement energy, effective reduction lactic acid content, avoids meta-bolites to the damage of body.
Summary of the invention
Technical problem to be solved by this invention is to provide that a kind of method is simple, the preparation method of the yak skin micromolecular collagen of the regulation and control human body energy metabolism of Be very effective.
For solving the problem, the preparation method of the yak skin micromolecular collagen of regulation and control human body energy of the present invention metabolism, comprises the following steps:
By fresh or through infiltrating, softening after the yak skin of Qinghai-Tibet Platean peculiar ox kind yak wildyak of drying clean up, then add by 2 ~ 3 times of its weight the alkali that mass concentration is 0.5 ~ 10%, be soak 2 ~ 72 hours under the condition of 20 ~ 38 DEG C in temperature, obtain the yak skin after losing hair or feathers;
(2) the yak skin after described depilation is added water by 3 ~ 5 times of its weight, be soak dealkalize under the condition of 20 ~ 30 DEG C, repeat 1 ~ 4 time, each 10 ~ 30 minutes in water temperature, then clear water rinses, and cleans remaining soft flocks, and measures pH value, obtains the yak skin of dealkalize depilation;
(3) the yak skin that described dealkalize is lost hair or feathers is cut into the fritter of 3 ~ 5cm, then add the water of its weight 5 ~ 20 times, be the skin 2 ~ 3 times of scalding under the condition of 40 ~ 60 DEG C in temperature, obtain the yak skin after degreasing;
(4) the water of the yak skin after described degreasing by its quality 8 ~ 12 times is extracted, filter after 2 ~ 10 hours, discard residue, obtain filtrate;
(5), after described filtrate being diluted with 5 ~ 50 times of water of its volume, be separated under the condition of 30 ~ 50 DEG C in separation temperature, obtain the former protein solution of the micromolecular Yak-skin Gelatin of 4Kda;
(6), by concentrated, dry for micromolecular for described 4Kda Yak-skin Gelatin former protein solution warp, obtain yak skin micromolecular collagen.
Described step (1) in alkali refer to one in sodium hydroxide solution, potassium hydroxide solution, sodium hydrogen carbonate solution.
Described step (5) in filtrate be separated in centrifugal, membrane sepn, dialysis process any one.
Described step (6) in concentratedly refer to concentrating under reduced pressure, dry up concentrated in any one.
Described step (6) in drying conditions be any one in spraying dry, lyophilize, microwave drying, oven drying.
The present invention compared with prior art has the following advantages:
1, the present invention is with yak skin for raw material, by alkaline process depilation process yak skin, obtains micromolecular collagen through extraction and isolation.The former molecular weight of albumen distribution of the Yak-skin Gelatin adopting the method obtained is concentrated, and its molecular weight is at 4KDa, and granularity is 2.9nm, and content is greater than more than 99.9% (see table 1, table 2, Fig. 1, Fig. 2).
Table 1: small molecules collagen DLS Dynamic Light Scattering Determination result
Note: Item is for carry out mensuration number of times to same batch sample.
Table 2 micromolecular collagen particle diameter peak Distribution
2, pharmacodynamic experiment is carried out to the yak skin micromolecular collagen of gained of the present invention:
(1) animal swimming with a load attached to the body experimental result:
Select two dosage (200mg/kg/d, 1.0g/kg/d) groups to carry out, healthy KM mouse 30 is divided into 3 groups at random, male and female half and half, blank group presses 0.2ml/10g gavage pure water, successive administration seven days.Before and after administration, the body weight of mouse, swimming time compare analysis, observe the impact (see table 3) of different dosing dosage mouse swimming time and body weight.
The impact that table 3 different dosing dosage is tested mice burden swimming
Compare with blank group: * P<0.05; * P<0.01.
(2) Biochemical Indexes:
By healthy KM mouse 30, be divided into 3 groups at random, male and female half and half, two administration groups (low, high), blank group, administration seven days.Detect the content (see table 4) of total protein (TP), serum lactic dehydrogenase (LDH), lactic acid (LA), liver glycogen, Body development.
Table 4 administration after seven days on the impact of blood biochemistry index
Compare with blank group: * P<0.05; * P<0.01.
The antifatigue experiment of different dosing dosage, can extend the swimming time of mouse, the liver Body development deposit of low, high 2 the dosed administration group mouse of the mensuration of biochemical indicator has significant lifting.Because when mouse exhausted movemeat, glycogen rapidly for mouse has carried out energy supply, thus makes the swimming time of mouse obtain growth (Coyle, Cogganetal.1986) as a kind of energy matter.
Under the administration of small-molecular-weight collagen protein high dosage, obviously reduce LDH and CK content, there is significance effect.Mouse is transformed into margin of energy material as glycogen and fat etc. through digesting and assimilating by body, and therefore in violent type exercise fatigue situation, margin of energy abundance but can not be supplied (Holloszy, Kohrtetal.1998) in time; And low molecular weight range is converted to some in vivo and participates in human body energy metabolic processes required enzyme or participate in intermediate product anabolic process going after being digested and assimilated by body; thus make body and energy requirement at once can effective supply energy, effectively reduce the damage that " the tired factor " lactic acid content protection body causes from meta-bolites.
Therefore, the yak skin micromolecular collagen of gained of the present invention has margin of energy function, and when needing, instant energy starts, and has removing function to meta-bolites lactic acid.
3, any chemical is not added in leaching process of the present invention, Be very effective, human energy metabolism can be regulated, energy can be provided effectively in time when body requirement energy, effective reduction lactic acid content, avoid meta-bolites to the damage of body, can be used for producing food, healthcare products, medicine and producing various solid preparation and liquid preparation, be especially applicable to the large crowd of labour intensity and eat.
Accompanying drawing explanation
Below in conjunction with accompanying drawing, the specific embodiment of the present invention is described in further detail.
Fig. 1 is the percentage composition distribution of micromolecular collagen of the present invention.
Fig. 2 is micromolecular collagen particle size content of the present invention distribution.
Embodiment
embodiment 1the preparation method of the yak skin micromolecular collagen of regulation and control human body energy metabolism, comprises the following steps:
(1) fresh yak skin is cleaned up, then add by 2 times of its weight the sodium hydroxide solution that mass concentration is 0.5%, be soak 2 hours under the condition of 38 DEG C in temperature, obtain the yak skin after losing hair or feathers.
(2) the yak skin after depilation is added water by 3 times of its weight, be soak dealkalize under the condition of 30 DEG C, repeat 2 times, each 30 minutes in water temperature, then clear water rinses, and cleans remaining soft flocks, and measures pH value, obtains the yak skin of dealkalize depilation.
(3) the yak skin that dealkalize is lost hair or feathers is cut into the fritter of 5cm, then add the water of its weight 20 times, be the skin 2 times of scalding under the condition of 60 DEG C in temperature, obtain the yak skin after degreasing.
(4) the water of the yak skin after degreasing by its quality 8 times is extracted, filter after 10 hours, discard residue, obtain filtrate.
By filtrate with after 5 times of water dilution of its volume, be under the condition of 30 DEG C in separation temperature, adopt aperture 10, molecular weight cut-off be 500 dialysis tubing carry out dialysis separation, obtain the former protein solution of the micromolecular Yak-skin Gelatin of 4Kda.
By the former protein solution of micromolecular for 4Kda Yak-skin Gelatin after concentrated, lyophilize, obtain yak skin micromolecular collagen.
Wherein: concentrate and refer to concentrating under reduced pressure, its condition is vacuum tightness 0.07MPa, temperature 50 C.Lyophilize condition refers to temperature-25 DEG C of precoolings 25 minutes ,-45 DEG C, dry 60 hours of 45MPa.
embodiment 2the preparation method of the yak skin micromolecular collagen of regulation and control human body energy metabolism, comprises the following steps:
(1) the yak skin of the Qinghai-Tibet Platean peculiar ox kind yak wildyak of the drying after infiltrating, softening is cleaned up, then add by 3 times of its weight the potassium hydroxide solution that mass concentration is 8%, be soak 50 hours under the condition of 25 DEG C in temperature, obtain the yak skin after losing hair or feathers.
(2) the yak skin after depilation is added water by 5 times of its weight, be soak dealkalize under the condition of 30 DEG C, repeat 3 times, each 30 minutes in water temperature, then clear water rinses, and cleans remaining soft flocks, and measures pH value, obtains the yak skin of dealkalize depilation.
(3) the yak skin that dealkalize is lost hair or feathers is cut into the fritter of 5cm, then add the water of its weight 10 times, be the skin 3 times of scalding under the condition of 50 DEG C in temperature, obtain the yak skin after degreasing.
(4) the water of the yak skin after degreasing by its quality 10 times is extracted, filter after 2 hours, discard residue, obtain filtrate.
By filtrate with after 25 times of water dilution of its volume, be 50 DEG C in separation temperature, under pressure is the condition of 3.5KPa, adopts 100K cellulose acetate film film to be separated, obtain the former protein solution of the micromolecular Yak-skin Gelatin of 4Kda.
By the former protein solution of micromolecular for 4Kda Yak-skin Gelatin after concentrated, spraying dry, obtain yak skin micromolecular collagen.
Wherein: concentrated referring to dries up concentrated, and its condition is temperature 15 DEG C, concentration time 60 hours.Spray-dired condition refers to mist flow 23ml/min, feeding temperature 40 DEG C, inlet temperature 175 DEG C.
embodiment 3the preparation method of the yak skin micromolecular collagen of regulation and control human body energy metabolism, comprises the following steps:
(1) fresh yak skin is cleaned up, then add by 2 times of its weight the sodium hydroxide solution that mass concentration is 10%, be soak 72 hours under the condition of 30 DEG C in temperature, obtain the yak skin after losing hair or feathers.
(2) the yak skin after depilation is added water by 5 times of its weight, be soak dealkalize under the condition of 20 DEG C, repeat 4 times, each 10 minutes in water temperature, then clear water rinses, and cleans remaining soft flocks, and measures pH value, obtains the yak skin of dealkalize depilation.
(3) the yak skin that dealkalize is lost hair or feathers is cut into the fritter of 4cm, then add the water of its weight 10 times, be the skin 3 times of scalding under the condition of 40 DEG C in temperature, obtain the yak skin after degreasing.
(4) the water of the yak skin after degreasing by its quality 9 times is extracted, filter after 8 hours, discard residue, obtain filtrate.
By filtrate with after 35 times of water dilution of its volume, in separation temperature be under the condition of 25 DEG C with 3000 revs/min of speed centrifugations 30 minutes, obtain the former protein solution of the micromolecular Yak-skin Gelatin of 4Kda.
By the former protein solution of micromolecular for 4Kda Yak-skin Gelatin after concentrated, microwave drying to constant weight, obtain yak skin micromolecular collagen.
Wherein: concentrate and refer to concentrating under reduced pressure, its condition is vacuum tightness 0.07MPa, temperature 50 C.The condition of microwave drying refers to power 1000W, 25 minutes time of drying.
embodiment 4the preparation method of the yak skin micromolecular collagen of regulation and control human body energy metabolism, comprises the following steps:
(1) fresh yak skin is cleaned up, then add by 3 times of its weight the sodium hydrogen carbonate solution that mass concentration is 7%, be soak 55 hours under the condition of 20 DEG C in temperature, obtain the yak skin after losing hair or feathers.
(2) the yak skin after depilation is added water by 4 times of its weight, be soak dealkalize under the condition of 25 DEG C, repeat 1 time, each 20 minutes in water temperature, then clear water rinses, and cleans remaining soft flocks, and measures pH value, obtains the yak skin of dealkalize depilation.
(3) the yak skin that dealkalize is lost hair or feathers is cut into the fritter of 3cm, then add the water of its weight 15 times, be the skin 2 times of scalding under the condition of 55 DEG C in temperature, obtain the yak skin after degreasing.
(4) the water of the yak skin after degreasing by its quality 12 times is extracted, filter after 4 hours, discard residue, obtain filtrate.
By filtrate with after 50 times of water dilution of its volume, be 50 DEG C in separation temperature, under pressure is the condition of 3.5KPa, adopts 100K cellulose acetate film film to be separated, obtain the former protein solution of the micromolecular Yak-skin Gelatin of 4Kda.
By the former protein solution of micromolecular for 4Kda Yak-skin Gelatin after concentrating, being dried to constant weight, obtain yak skin micromolecular collagen.
Wherein: concentrate and refer to concentrating under reduced pressure, its condition is vacuum tightness 0.07MPa, temperature 50 C.Dry condition refers to 40 ~ 50 DEG C of thermostatic drying chambers dry 25 hours.
embodiment 5the preparation method of the yak skin micromolecular collagen of regulation and control human body energy metabolism, comprises the following steps:
(1) the yak skin of the Qinghai-Tibet Platean peculiar ox kind yak wildyak of the drying after infiltrating, softening is cleaned up, then add by 2.5 times of its weight the sodium hydrogen carbonate solution that mass concentration is 5%, be soak 12 hours under the condition of 28 DEG C in temperature, obtain the yak skin after losing hair or feathers.
(2) the yak skin after depilation is added water by 3.5 times of its weight, be soak dealkalize under the condition of 27 DEG C, repeat 2 times, each 25 minutes in water temperature, then clear water rinses, and cleans remaining soft flocks, and measures pH value, obtains the yak skin of dealkalize depilation.
(3) the yak skin that dealkalize is lost hair or feathers is cut into the fritter of 3cm, then add the water of its weight 5 times, be the skin 2 times of scalding under the condition of 45 DEG C in temperature, obtain the yak skin after degreasing.
(4) the water of the yak skin after degreasing by its quality 9 times is extracted, filter after 5 hours, discard residue, obtain filtrate.
By filtrate with after 50 times of water dilution of its volume, be 50 DEG C in separation temperature, under pressure is the condition of 3.5KPa, adopts 100K cellulose acetate film film to be separated, obtain the former protein solution of the micromolecular Yak-skin Gelatin of 4Kda.
By the former protein solution of micromolecular for 4Kda Yak-skin Gelatin after concentrating, being dried to constant weight, obtain yak skin micromolecular collagen.
Wherein: concentrate and refer to concentrating under reduced pressure, its condition is vacuum tightness 0.07MPa, temperature 50 C.Dry condition refers to 40 ~ 50 DEG C of thermostatic drying chambers dry 25 hours.
Claims (5)
1. regulate and control the preparation method of the yak skin micromolecular collagen of human body energy metabolism, comprise the following steps:
By fresh or through infiltrating, softening after the yak skin of Qinghai-Tibet Platean peculiar ox kind yak wildyak of drying clean up, then add by 2 ~ 3 times of its weight the alkali that mass concentration is 0.5 ~ 10%, be soak 2 ~ 72 hours under the condition of 20 ~ 38 DEG C in temperature, obtain the yak skin after losing hair or feathers;
(2) the yak skin after described depilation is added water by 3 ~ 5 times of its weight, be soak dealkalize under the condition of 20 ~ 30 DEG C, repeat 1 ~ 4 time, each 10 ~ 30 minutes in water temperature, then clear water rinses, and cleans remaining soft flocks, and measures pH value, obtains the yak skin of dealkalize depilation;
(3) the yak skin that described dealkalize is lost hair or feathers is cut into the fritter of 3 ~ 5cm, then add the water of its weight 5 ~ 20 times, be the skin 2 ~ 3 times of scalding under the condition of 40 ~ 60 DEG C in temperature, obtain the yak skin after degreasing;
(4) the water of the yak skin after described degreasing by its quality 8 ~ 12 times is extracted, filter after 2 ~ 10 hours, discard residue, obtain filtrate;
(5), after described filtrate being diluted with 5 ~ 50 times of water of its volume, be separated under the condition of 30 ~ 50 DEG C in separation temperature, obtain the former protein solution of the micromolecular Yak-skin Gelatin of 4Kda;
(6), by concentrated, dry for micromolecular for described 4Kda Yak-skin Gelatin former protein solution warp, obtain yak skin micromolecular collagen.
2. the preparation method of the yak skin micromolecular collagen of regulation and control human body energy as claimed in claim 1 metabolism, is characterized in that: described step (1) in alkali refer to one in sodium hydroxide solution, potassium hydroxide solution, sodium hydrogen carbonate solution.
3. the preparation method of the yak skin micromolecular collagen of regulation and control human body energy as claimed in claim 1 metabolism, is characterized in that: described step (5) in filtrate be separated in centrifugal, membrane sepn, dialysis process any one.
4. the preparation method of the yak skin micromolecular collagen of regulation and control human body energy as claimed in claim 1 metabolism, is characterized in that: described step (6) in concentratedly refer to concentrating under reduced pressure, dry up concentrated in any one.
5. the preparation method of the yak skin micromolecular collagen of regulation and control human body energy as claimed in claim 1 metabolism, is characterized in that: described step (6) in drying conditions be any one in spraying dry, lyophilize, microwave drying, oven drying.
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| CN107362356A (en) * | 2017-03-17 | 2017-11-21 | 中国科学院西北高原生物研究所 | A kind of Yak-skin Gelatin original protein product with hemostasia effect and preparation method thereof |
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| CN102614499A (en) * | 2012-03-15 | 2012-08-01 | 中国科学院西北高原生物研究所 | Yak skin collagen compound traditional Tibetan medicine with anti-fatigue effect and preparation process thereof |
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| CN102614499A (en) * | 2012-03-15 | 2012-08-01 | 中国科学院西北高原生物研究所 | Yak skin collagen compound traditional Tibetan medicine with anti-fatigue effect and preparation process thereof |
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| CN107362356A (en) * | 2017-03-17 | 2017-11-21 | 中国科学院西北高原生物研究所 | A kind of Yak-skin Gelatin original protein product with hemostasia effect and preparation method thereof |
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Address after: 810001 Xiguan Street, Xining, Xining, Qinghai Co-patentee after: Qinghai Provincial Institute of drug control and inspection Patentee after: Northwest Institute of Plateau Biology, Chinese Academy of Sciences Address before: 810001 Xiguan Street, Xining, Xining, Qinghai Co-patentee before: Qinghai Province food and medicine inspection institute Patentee before: Northwest Institute of Plateau Biology, Chinese Academy of Sciences |
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Effective date of registration: 20180903 Address after: 810000 22 Wei three road, Chengbei District, Xining, Qinghai (Qinghai Biotechnology Industrial Park) Patentee after: Qinghai Qiying Sino-tibetan Biologic Pharmacy Co., Ltd. Address before: 810001 Xiguan Street, Xining, Xining, Qinghai Co-patentee before: Qinghai Provincial Institute of drug control and inspection Patentee before: Northwest Institute of Plateau Biology, Chinese Academy of Sciences |