CN103820517A - Preparation method for yak skin micromolecular collagen capable of regulating human body energy metabolism - Google Patents
Preparation method for yak skin micromolecular collagen capable of regulating human body energy metabolism Download PDFInfo
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Abstract
The invention relates to a preparation method for yak skin micromolecular collagen capable of regulating human body energy metabolism. The method comprises the following steps: (1), cleaning up fresh or soaked and softened dry yak skin of a special yak wild yak in Qinghai-Tibet Plateau, and then adding alkali for soaking, so as to obtain hair removed yak skin; (2), adding water for soaking the hair removed yak skin, dealkalizing, and then cleaning with clean water, so as to obtain dealkalized hair removed yak skin; (3), shearing the dealkalized hair removed yak skin into small pieces, and then adding water for blanching the skin pieces, so as to obtain degreased yak skin; (4),extracting the degreased yak skin with water, and then filtering for removing residues, so as to obtain a filtrate; (5), diluting the filtrate with water, and then separating, so as to obtain a 4Kda micromolecular yak skin collagen solution; (6), concentrating and drying the 4Kda micromolecular yak skin collagen solution, so as to obtain the yak skin micromolecular collagen. The method is simple and remarkable in effect, and the prepared yak skin collagen is concentrated in molecular weight distribution, and has the function of regulating human body energy metabolism.
Description
Technical field
The present invention relates to a kind of preparation method of collagen protein, relate in particular to the preparation method of the yak skin micromolecular collagen of regulation and control human body energy metabolism.
Background technology
Sugar, fat and the protein of the required energy derive of body in food, these nutritive ingredients are transformed into absorbable small molecules nutritive substance and are absorbed through digestion.Human body nearly 16% is protein, and in protein, has 35 ~ 45% collagen protein, and there are skin, muscle, bone, tooth etc. in the main position existing, 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, reduction and oxidizing reaction between catalysis pyruvic acid and lactic acid, pyruvic acid is reduced to lactic acid in the time that glycolysis-speed exceedes aerobic metabolism speed, is the important enzyme of involved in sugar anaerobic glycolysis and glyconeogenesis.
Creatine kinase (CK) is present in the cytoplasm and plastosome of the tissues such as heart, muscle and brain; the important kinases that and intracellular energy running, Muscle contraction, ATP regeneration have direct relation, it reversibly the phosphoryl that turns between catalysis creatine and ATP react.
Triphosaden (ATP) is directly supplied with the material of energy, PCr is main margin of energy material, under physiological conditions, ATP can be shifted and form PCr from mitochondrial inner membrane by mitochondrial creatine kinase, and PCr transfers ATP to for utilization through the catalysis of kytoplasm CK isozyme again at need energy position.
Yak is one of ancient and original ox kind of China, is the ox kind of unique procreation of 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, its unique heritability and the adaptive faculty to high and cold anoxic severe environment are formed, there is extremely strong anti-adversity ability, resistance against diseases, tolerance and adaptability, can adapt to the harsh high and cold severe ecotopes such as the immalleable high height above sea level of general domestic animal, high anoxic.Yak body contains the biologically active substance of many antifatigues, anti-hypoxia.From yak skin, extract the micromolecular collagen of pure natural, after being absorbed by body, be converted to and participate in human body energy metabolic process required enzyme or participate in intermediate product anabolic process, in the time of body requirement energy, can provide effectively in time energy, effectively reduce lactic acid content, avoid the damage of meta-bolites to body.
Summary of the invention
Technical problem to be solved by this invention is to provide the preparation method of the yak skin micromolecular collagen that a kind of method is simple, effect regulates and controls human body energy metabolism significantly.
For addressing the above 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:
(1) the yak skin of fresh or dry Qinghai-Tibet Platean peculiar ox kind yak wild yak through infiltrating, after softening is cleaned up, then adding mass concentration by 2 ~ 3 times of its weight is 0.5 ~ 10% alkali, be to soak 2 ~ 72 hours under the condition of 20 ~ 38 ℃ in temperature, the yak skin after being lost hair or feathers;
(2) the yak skin after described depilation being added to water by 3 ~ 5 times of its weight, is to soak dealkalize under the condition of 20 ~ 30 ℃ in water temperature, repeats 1 ~ 4 time, and each 10 ~ 30 minutes, then clear water rinsed, and cleaned remaining soft flocks, and measured pH value, obtained the yak skin that dealkalize is lost hair or feathers;
(3) the yak skin of described dealkalize depilation being cut into the fritter of 3 ~ 5cm, then add the water of 5 ~ 20 times of its weight, is the skin 2 ~ 3 times of scalding under the condition of 40 ~ 60 ℃ in temperature, obtains the yak skin after degreasing;
(4) the yak skin after described degreasing is extracted with the water of 8 ~ 12 times of its quality, after 2 ~ 10 hours, filter, discard residue, obtain filtrate;
(5) by described filtrate after 5 ~ 50 times of water dilution with its volume, be to separate under the condition of 30 ~ 50 ℃ in separation temperature, obtain the former protein solution of the micromolecular Yak-skin Gelatin of 4Kda;
(6) by concentrated, dry former micromolecular described 4Kda Yak-skin Gelatin protein solution warp, obtain yak skin micromolecular collagen.
The alkali of described step in (1) refers to the one in sodium hydroxide solution, potassium hydroxide solution, sodium hydrogen carbonate solution.
That the filtrate of described step in is (5) separated into is centrifugal, any one in membrane sepn, dialysis process.
Concentrated in (6) of described step refer to concentrating under reduced pressure, dry up concentrated in any one.
The drying conditions of described step in is (6) that spraying is dry, any one in lyophilize, microwave drying, oven drying.
The present invention compared with prior art has the following advantages:
1, the present invention, take yak skin as raw material, is lost hair or feathers and is processed yak skin by alkaline process, obtains micromolecular collagen through extracting to separate.The former molecular weight of albumen of Yak-skin Gelatin that adopts the method to make distributes and concentrates, and its molecular weight is at 4KDa, and granularity is 2.9nm, and content is greater than more than 99.9% (referring to table 1, table 2, Fig. 1, Fig. 2).
Table 1: small molecules collagen DLS Dynamic Light Scattering Determination result
Note: Item is for to measure number of times to same batch sample.
Table 2 micromolecular collagen particle diameter peak Distribution
2, the yak skin micromolecular collagen of gained of the present invention is carried out to pharmacodynamic experiment:
(1) animal swimming with a load attached to the body experimental result:
Select two dosages (200 mg/kg/d, 1.0 g/kg/d) group to carry out, 30 of healthy KM mouse are divided into 3 groups at random, male and female half and half, blank group is by 0.2 ml/10 g gavage pure water, successive administration seven days.Before and after administration, body weight, the swimming time of mouse compare analysis, observe the impact (referring to table 3) of different dosing dosage mouse swimming time and body weight.
The impact of table 3 different dosing dosage on mice burden swimming experiment
With the comparison of blank group: * P<0.05; * P<0.01.
(2) Biochemical Indexes:
By 30 of healthy KM mouse, be divided at random 3 groups, half and half, two administration group of male and female (low, height), blank group, administration seven days.Detect the content (referring to table 4) of total protein (TP), serum lactic dehydrogenase (LDH), lactic acid (LA), liver glycogen, inose unit.
Table 4 administration is the impact on blood biochemistry index after seven days
With the comparison of blank group: * P<0.05; * P<0.01.
The antifatigue experiment of different dosing dosage, can extend the swimming time of mouse, and the liver inose unit deposit of low, high 2 the dosed administration group mouse of mensuration of biochemical indicator has significant lifting.Because exhaust when motion in mouse power, glycogen as a kind of energy matter rapidly for mouse has carried out energy supply, thereby make the swimming time of mouse obtain growth (Coyle, Coggan et al. 1986).
Under the administration of small molecules amount collagen protein high dosage, obviously reduce LDH and CK content, there is significance effect.Mouse transforms 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, Kohrt et al. 1998) in time; And small molecules weight range is converted in vivo some 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; thereby make body and energy requirement at once can effective supply energy, effectively reduce " the tired factor " lactic acid content protection body and avoid the damage that meta-bolites causes.
Therefore, the yak skin micromolecular collagen of gained of the present invention has margin of energy function, and while needs, instant energy starts, and meta-bolites lactic acid is had to removing function.
3, in leaching process of the present invention, do not add any chemical, effect is remarkable, can regulate human energy metabolism, in the time of body requirement energy, can provide effectively in time energy, effectively reduce lactic acid content, avoid the damage of meta-bolites to body, can be used for producing food, healthcare products, medicine and produce various solid preparations and liquid preparation, be especially applicable to the crowd that labour intensity is large edible.
Accompanying drawing explanation
Below in conjunction with accompanying drawing, the specific embodiment of the present invention is described in further detail.
Fig. 1 is that the percentage composition of micromolecular collagen of the present invention distributes.
Fig. 2 is micromolecular collagen particle diameter content distribution of the present invention.
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 adding mass concentration by 2 times of its weight is 0.5% sodium hydroxide solution, is to soak 2 hours under the condition of 38 ℃ the yak skin after being lost hair or feathers in temperature.
(2) the yak skin after depilation being added to water by 3 times of its weight, is to soak dealkalize under the condition of 30 ℃ in water temperature, repeats 2 times, and each 30 minutes, then clear water rinsed, and cleaned remaining soft flocks, and measured pH value, obtained the yak skin that dealkalize is lost hair or feathers.
(3) the yak skin of dealkalize depilation being cut into the fritter of 5cm, then add the water of 20 times of its weight, is the skin 2 times of scalding under the condition of 60 ℃ in temperature, obtains the yak skin after degreasing.
(4) the yak skin after degreasing is extracted with the water of 8 times of its quality, after 10 hours, filter, discard residue, obtain filtrate.
(5) by filtrate after the 5 times of water dilution with its volume, be, under the condition of 30 ℃, to adopt aperture 10 in separation temperature, the dialysis tubing that molecular weight cut-off is 500 separation of dialysing, obtains the former protein solution of the micromolecular Yak-skin Gelatin of 4Kda.
By the former protein solution of micromolecular 4Kda Yak-skin Gelatin after concentrated, lyophilize, obtain yak skin micromolecular collagen.
Wherein: concentrate and refer to that concentrating under reduced pressure, its condition are vacuum tightness 0.07MPa, temperature 50 C.Lyophilize condition refers to temperature-25 ℃ precooling 25 minutes, dry 60 hours of-45 ℃, 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 dry Qinghai-Tibet Platean peculiar ox kind yak wild yak after infiltrating, softening is cleaned up, then adding mass concentration by 3 times of its weight is 8% potassium hydroxide solution, be to soak 50 hours under the condition of 25 ℃ in temperature, the yak skin after being lost hair or feathers.
(2) the yak skin after depilation being added to water by 5 times of its weight, is to soak dealkalize under the condition of 30 ℃ in water temperature, repeats 3 times, and each 30 minutes, then clear water rinsed, and cleaned remaining soft flocks, and measured pH value, obtained the yak skin that dealkalize is lost hair or feathers.
(3) the yak skin of dealkalize depilation being cut into the fritter of 5cm, then add the water of 10 times of its weight, is the skin 3 times of scalding under the condition of 50 ℃ in temperature, obtains the yak skin after degreasing.
(4) the yak skin after degreasing is extracted with the water of 10 times of its quality, after 2 hours, filter, discard residue, obtain filtrate.
(5) by filtrate after the 25 times of water dilution with its volume, be under 50 ℃, the pressure condition that is 3.5KPa in separation temperature, adopt 100K cellulose acetate film film to separate, obtain the former protein solution of the micromolecular Yak-skin Gelatin of 4Kda.
By the former protein solution of micromolecular 4Kda Yak-skin Gelatin after concentrated, spraying is dry, obtain yak skin micromolecular collagen.
Wherein: concentrated referring to dries up concentratedly, and its condition is 15 ℃ of temperature, concentration time 60 hours.Spray-dired condition refers to mist flow 23ml/min, 40 ℃ of feeding temperatures, 175 ℃ of inlet temperature.
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 adding mass concentration by 2 times of its weight is 10% sodium hydroxide solution, is to soak 72 hours under the condition of 30 ℃ the yak skin after being lost hair or feathers in temperature.
(2) the yak skin after depilation being added to water by 5 times of its weight, is to soak dealkalize under the condition of 20 ℃ in water temperature, repeats 4 times, and each 10 minutes, then clear water rinsed, and cleaned remaining soft flocks, and measured pH value, obtained the yak skin that dealkalize is lost hair or feathers.
(3) the yak skin of dealkalize depilation being cut into the fritter of 4cm, then add the water of 10 times of its weight, is the skin 3 times of scalding under the condition of 40 ℃ in temperature, obtains the yak skin after degreasing.
(4) the yak skin after degreasing is extracted with the water of 9 times of its quality, after 8 hours, filter, discard residue, obtain filtrate.
(5) by filtrate after the 35 times of water dilution with its volume, in separation temperature be under the condition of 25 ℃ 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 4Kda Yak-skin Gelatin through concentrated, microwave drying to constant weight, obtain yak skin micromolecular collagen.
Wherein: concentrate and refer to that concentrating under reduced pressure, its condition are 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 adding mass concentration by 3 times of its weight is 7% sodium hydrogen carbonate solution, is to soak 55 hours under the condition of 20 ℃ the yak skin after being lost hair or feathers in temperature.
(2) the yak skin after depilation being added to water by 4 times of its weight, is to soak dealkalize under the condition of 25 ℃ in water temperature, repeats 1 time, and each 20 minutes, then clear water rinsed, and cleaned remaining soft flocks, and measured pH value, obtained the yak skin that dealkalize is lost hair or feathers.
(3) the yak skin of dealkalize depilation being cut into the fritter of 3cm, then add the water of 15 times of its weight, is the skin 2 times of scalding under the condition of 55 ℃ in temperature, obtains the yak skin after degreasing.
(4) the yak skin after degreasing is extracted with the water of 12 times of its quality, after 4 hours, filter, discard residue, obtain filtrate.
(5) by filtrate after the 50 times of water dilution with its volume, be under 50 ℃, the pressure condition that is 3.5KPa in separation temperature, adopt 100K cellulose acetate film film to separate, obtain the former protein solution of the micromolecular Yak-skin Gelatin of 4Kda.
By the former protein solution of micromolecular 4Kda Yak-skin Gelatin through concentrating, be dried to after constant weight, obtain yak skin micromolecular collagen.
Wherein: concentrate and refer to that concentrating under reduced pressure, its condition are vacuum tightness 0.07MPa, temperature 50 C.Dry condition refers to 40 ~ 50 ℃ 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 dry Qinghai-Tibet Platean peculiar ox kind yak wild yak after infiltrating, softening is cleaned up, then adding mass concentration by 2.5 times of its weight is 5% sodium hydrogen carbonate solution, be to soak 12 hours under the condition of 28 ℃ in temperature, the yak skin after being lost hair or feathers.
(2) the yak skin after depilation being added to water by 3.5 times of its weight, is to soak dealkalize under the condition of 27 ℃ in water temperature, repeats 2 times, and each 25 minutes, then clear water rinsed, and cleaned remaining soft flocks, and measured pH value, obtained the yak skin that dealkalize is lost hair or feathers.
(3) the yak skin of dealkalize depilation being cut into the fritter of 3cm, then add the water of 5 times of its weight, is the skin 2 times of scalding under the condition of 45 ℃ in temperature, obtains the yak skin after degreasing.
(4) the yak skin after degreasing is extracted with the water of 9 times of its quality, after 5 hours, filter, discard residue, obtain filtrate.
(5) by filtrate after the 50 times of water dilution with its volume, be under 50 ℃, the pressure condition that is 3.5KPa in separation temperature, adopt 100K cellulose acetate film film to separate, obtain the former protein solution of the micromolecular Yak-skin Gelatin of 4Kda.
By the former protein solution of micromolecular 4Kda Yak-skin Gelatin through concentrating, be dried to after constant weight, obtain yak skin micromolecular collagen.
Wherein: concentrate and refer to that concentrating under reduced pressure, its condition are vacuum tightness 0.07MPa, temperature 50 C.Dry condition refers to 40 ~ 50 ℃ of thermostatic drying chambers dry 25 hours.
Claims (5)
1. the 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 fresh or dry Qinghai-Tibet Platean peculiar ox kind yak wild yak through infiltrating, after softening is cleaned up, then adding mass concentration by 2 ~ 3 times of its weight is 0.5 ~ 10% alkali, be to soak 2 ~ 72 hours under the condition of 20 ~ 38 ℃ in temperature, the yak skin after being lost hair or feathers;
(2) the yak skin after described depilation being added to water by 3 ~ 5 times of its weight, is to soak dealkalize under the condition of 20 ~ 30 ℃ in water temperature, repeats 1 ~ 4 time, and each 10 ~ 30 minutes, then clear water rinsed, and cleaned remaining soft flocks, and measured pH value, obtained the yak skin that dealkalize is lost hair or feathers;
(3) the yak skin of described dealkalize depilation being cut into the fritter of 3 ~ 5cm, then add the water of 5 ~ 20 times of its weight, is the skin 2 ~ 3 times of scalding under the condition of 40 ~ 60 ℃ in temperature, obtains the yak skin after degreasing;
(4) the yak skin after described degreasing is extracted with the water of 8 ~ 12 times of its quality, after 2 ~ 10 hours, filter, discard residue, obtain filtrate;
(5) by described filtrate after 5 ~ 50 times of water dilution with its volume, be to separate under the condition of 30 ~ 50 ℃ in separation temperature, obtain the former protein solution of the micromolecular Yak-skin Gelatin of 4Kda;
(6) by concentrated, dry former micromolecular described 4Kda Yak-skin Gelatin 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: the alkali of described step in (1) refers to the 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: the filtrate of described step in is (5) separated into any one in centrifugal, membrane sepn, dialysis process.
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: concentrated in (6) of described step 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: the drying conditions of described step in is (6) that spraying is dry, any one in lyophilize, microwave drying, oven drying.
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| CN105596261A (en) * | 2016-01-29 | 2016-05-25 | 中国科学院西北高原生物研究所 | Yak skin collagen product with functions of maintaining beauty and keeping young and preparation method of yak skin collagen product |
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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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| Publication number | Priority date | Publication date | Assignee | Title |
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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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| CN104000848A (en) * | 2014-06-05 | 2014-08-27 | 中国科学院西北高原生物研究所 | Method for improving excessive internal heat caused by taking Maoxijiao granules |
| CN105596261A (en) * | 2016-01-29 | 2016-05-25 | 中国科学院西北高原生物研究所 | Yak skin collagen product with functions of maintaining beauty and keeping young and preparation method of yak skin collagen product |
| CN105662998A (en) * | 2016-01-29 | 2016-06-15 | 中国科学院西北高原生物研究所 | Yak hide collagen mask with ultraviolet ray resistance and preparation method |
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| CN109295145A (en) * | 2018-10-31 | 2019-02-01 | 西藏央金生态农牧科技有限公司 | A kind of preparation method of Ultra-low molecular weight yak collagen peptide |
| CN110229860A (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 Skin small-molecular peptides preparation method promoting Marrow Stromal Cells in Proliferation |
| CN110241161A (en) * | 2019-06-21 | 2019-09-17 | 中国科学院西北高原生物研究所 | A kind of yak skin small-molecular peptides preparation method enhancing marrow stromal cell ability |
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