CN106939325A - A kind of preparation method of ox bone collagen albumen - Google Patents
A kind of preparation method of ox bone collagen albumen Download PDFInfo
- Publication number
- CN106939325A CN106939325A CN201610001744.4A CN201610001744A CN106939325A CN 106939325 A CN106939325 A CN 106939325A CN 201610001744 A CN201610001744 A CN 201610001744A CN 106939325 A CN106939325 A CN 106939325A
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- Prior art keywords
- bone
- collagen
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- digestion reaction
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- 210000000988 Bone and Bones Anatomy 0.000 title claims abstract description 91
- 102000008186 Collagen Human genes 0.000 title claims abstract description 47
- 108010035532 Collagen Proteins 0.000 title claims abstract description 47
- 229960005188 collagen Drugs 0.000 title claims abstract description 47
- 229920001436 collagen Polymers 0.000 title claims abstract description 47
- 238000002360 preparation method Methods 0.000 title claims abstract description 16
- OKTJSMMVPCPJKN-UHFFFAOYSA-N carbon Chemical compound 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[C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 32
- 241000283690 Bos taurus Species 0.000 claims abstract description 24
- 239000000843 powder Substances 0.000 claims abstract description 24
- 238000004140 cleaning Methods 0.000 claims abstract description 17
- 238000001914 filtration Methods 0.000 claims abstract description 15
- 102000004190 Enzymes Human genes 0.000 claims abstract description 9
- 108090000790 Enzymes Proteins 0.000 claims abstract description 9
- 238000005238 degreasing Methods 0.000 claims abstract description 8
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O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 5
- 108090000284 Pepsin A Proteins 0.000 claims description 4
- 238000000498 ball milling Methods 0.000 claims description 4
- 238000009835 boiling Methods 0.000 claims description 4
- 239000008367 deionised water Substances 0.000 claims description 4
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 claims description 4
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- 230000015572 biosynthetic process Effects 0.000 description 2
- OYPRJOBELJOOCE-UHFFFAOYSA-N calcium Chemical compound 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[Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 2
- 239000011575 calcium Substances 0.000 description 2
- 229910052791 calcium Inorganic materials 0.000 description 2
- 238000004033 diameter control Methods 0.000 description 2
- 239000000835 fiber Substances 0.000 description 2
- 238000005755 formation reaction Methods 0.000 description 2
- 239000008187 granular material Substances 0.000 description 2
- 229910052500 inorganic mineral Inorganic materials 0.000 description 2
- -1 magnesium phosphorus Chemical compound 0.000 description 2
- 239000011707 mineral Substances 0.000 description 2
- 230000000050 nutritive Effects 0.000 description 2
- 238000010298 pulverizing process Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 241000272525 Anas platyrhynchos Species 0.000 description 1
- 102000016942 Elastin Human genes 0.000 description 1
- 108010014258 Elastin Proteins 0.000 description 1
- 210000000936 Intestines Anatomy 0.000 description 1
- 208000008466 Metabolic Disease Diseases 0.000 description 1
- 208000001132 Osteoporosis Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000005904 alkaline hydrolysis reaction Methods 0.000 description 1
- 230000003712 anti-aging Effects 0.000 description 1
- 239000000919 ceramic Substances 0.000 description 1
- 201000010251 cutis laxa Diseases 0.000 description 1
- 238000010612 desalination reaction Methods 0.000 description 1
- 229920002549 elastin Polymers 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000005416 organic matter Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 229910001220 stainless steel Inorganic materials 0.000 description 1
- 239000010935 stainless steel Substances 0.000 description 1
- 239000002351 wastewater Substances 0.000 description 1
- 238000004065 wastewater treatment Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
Abstract
The invention belongs to collagen preparation field, it discloses a kind of preparation method of ox bone collagen albumen, including processing step：Ox bone is cleaned, ox bone is crushed, degreasing, decalcification processing bovine bone powder are digested to bovine bone powder, destroy the enzyme treatment are carried out at 100 DEG C, filtering obtains collagen solution, and activated carbon decolorizing processing, concentration obtains collagen.Preparation technology of the present invention is simple, non-environmental-pollution, and production cost is low.
Description
Technical field
The present invention relates to collagen extractive technique field, more particularly to a kind of preparation method of ox bone collagen albumen.Background technology
Human body bone collagen metabolic disorder, is to cause the major reason of various osteopathy.It is to be not easy to effect a radical cure osteopathy with other methods if being unable to the albumen of collagen hyalronic.Human body bone is slightly made up of cartilage and bone.The collagen that wherein content of collagen is accounted in the 40% of cartilage matrix, os osseum in cartilage accounts for the 90%. of organic matter because than being described as os in os containing f.Bone road is just as a net for being full of duck eye.Firmly pin calcium and magnesium phosphorus and other mineral compositions.If without bone collagen, calcium and magnesium phosphorus and other mineral matters will not be evenly dispersed on bone, and bone road also would not be so hard, and what joint also will not be so is flexible.Intestines acid reason pain, joint will occur and fall pain and osteoporosis.
Bone collagen uses pharmagel for raw material substantially.Elastin is rich in bone collagen, for improving cutis laxa, the effect of anti-aging can become apparent from.Therefore no matter from technique or effect, pure bone collagen is more suitable for for absorption of human body.
The content of the invention
Based on above mentioned problem, problem to be solved by this invention is to provide a kind of simple to operate, non-environmental-pollution, and the preparation method of the ox bone collagen albumen of technique productions low cost.
Technical scheme is as follows：
A kind of preparation method of ox bone collagen albumen, comprises the following steps：
S1, cleaning：Half ox bone more than one full year of life, below two one full year of life is put into rinse bath, then toward adding clear water in rinse bath and flooding ox bone completely, ultrasonic agitation then is carried out 30 ~ 60 minutes to rinse bath；The step is repeated repeatedly, to remove the impurity on ox bone surface；It is preferred that the ox bone of an one full year of life, the collagen of this ox bone constitutes the slightly curved fiber of a branch of beam, mutually with last layer and next layer of formation right-angle array, and quantity is more, is of high nutritive value；
S2, drying：Clean ox bone is dried 12 ~ 24 hours at 60 ~ 80 DEG C；
S3, crushing：After with pulverizer, dry ox bone is crushed again on ball mill ball milling into the bovine bone powder that particle diameter is 5 ~ 200 microns；
S4, low temperature reflux degreasing 4 ~ 5 hours will be carried out under 60 DEG C of water bath conditions of bovine bone powder；It is preferred that reflux temperature is 80 ~ 100 DEG C；
S5, decalcification：The bovine bone powder of ungrease treatment is put into 10% hydrochloric acid solution and soaked 12 ~ 24 hours, decalcification processing is carried out, and a hydrochloric acid solution was changed every 3 hours, hydrochloric acid solution is cleaned up repeatedly with clear water again；
S6, enzymolysis：Bovine bone powder after decalcification is handled is poured into enzyme digestion reaction tank, and toward the deionized water that 0.1 ~ 0.5 times of bovine bone powder weight is added in enzyme digestion reaction tank, and add the acid protease of bovine bone powder weight 0.1 ~ 0.5%, then 30 ~ 50 DEG C are warming up to the progress of enzyme digestion reaction tank while stirring, enzymolysis 2 ~ 5 hours, while the pH value in controlled enzymatic hydrolysis retort is 2.5 ~ 4；
S7, go out enzyme：After enzymolysis is finished, enzyme digestion reaction tank is warming up to 100 DEG C, the slurry micro-boiling 5 ~ 10 minutes after enzymolysis is kept, acid protease is lost activity；
S8, filtering：After the enzyme that goes out is finished, enzyme digestion reaction tank is cooled to 50 ~ 60 DEG C, then will be filtered using vertical diatomite filtering machine, and be removed filter residue, obtain collagen solution；
S9, decolouring：Activated carbon is added into collagen solution and carries out decolorization, the addition of activated carbon is 0.1 ~ 0.5 times of collagen solution weight；
S10, concentration：The activated carbon after decolorization is filtered, and the filtrate obtained after filtering is passed through in batch (-type) single-action coiled vacuum concentration pan according to 550kg/h flow, and vacuum concentration processing is carried out at for 65 ~ 70 DEG C, moisture removal is removed, obtains solid matter；
S11, drying：The solid matter that vacuum concentration processing is obtained, which is placed in the drying box of air-dry machine, is dried processing；Wherein, the EAT of air-dry machine is 230 DEG C, and leaving air temp is 95 DEG C, obtains ox bone collagen albumen.
The preparation method of the ox bone collagen albumen, it is preferable that acid protease is pepsin, SUKAPro AC, 537 acid proteases.
The preparation method of the ox bone collagen albumen, it is preferable that activated carbon is granular active carbon, and particle diameter is 20 ~ 40 mesh.
The preparation method for the ox bone collagen albumen that the present invention is provided, has the following advantages that：
1st, the desalination degreasings such as acidolysis, alkaline hydrolysis need not be carried out using substantial amounts of acid, aqueous slkali, the waste water of exclusion is environmentally safe, meanwhile, and due to eliminating waste water treatment process, reduce production cost；
2nd, in enzymolysis process, the water of addition is few, and hydrolysis temperature is low, substantially reduces enzymolysis time, to obtain small-molecular-weight collagen, it is easy to absorb；
3rd, the product after enzyme digestion reaction, good filtration effect are filtered using vertical diatomite filtering machine, and the collagen obtained presses white solution purity；
4th, from half more than one full year of life two one full year of life following ox bone as the raw material for extracting bone collagen, this ox bone collagen albumen constitutes the slightly curved fiber of a branch of beam, and mutually with last layer and next layer of formation right-angle array, and quantity is more, is of high nutritive value.
Embodiment
Presently preferred embodiments of the present invention is described in further detail below.
Embodiment 1：
The step of preparation process of ox bone collagen albumen is as follows：
S1, cleaning：The ox bone of 5Kg half one full year of life oxen is put into rinse bath, then toward clear water is added in rinse bath, the bloodstain sticked on ox bone, viscous thing are rinsed repeatedly；Then in injected clear water and ox bone is flooded completely, then carrying out ultrasonic vibration to rinse bath stirs 30 minutes；Repeat the step 4-6 times, it is standby until ox bone surface clean is clean to remove the impurity on ox bone surface；
S2, drying：Clean ox bone is put into drying baker, is dried 12 hours under air atmosphere at 80 DEG C, in drying process, an ox bone was stirred every 2 hours, ox bone is gone moisture removal, uniform drying as far as possible；
S3, crushing：After with pulverizer, dry ox bone is crushed again on ball mill ball milling into the bovine bone powder that particle diameter is 5 microns；For ease of crushing, before crushing, ox bone first can be cut into sheet, depending on thickness is as needed, then carry out pulverization process with pulverizer；Ball mill is ground using agate ball, and sphere diameter control is at 5 ~ 20 millimeters；
S4, degreasing：Bovine bone powder is placed under 60 DEG C of water bath conditions and carries out low temperature reflux degreasing 4 hours, and reflux temperature control is at 80 DEG C or so；
S5, decalcification：Bovine bone powder after ungrease treatment is put into 10% hydrochloric acid solution and soaked 12 hours, decalcification processing is carried out, and a hydrochloric acid solution was changed every 3 hours, hydrochloric acid solution is cleaned up repeatedly with clear water again, it is complete after filter moisture；
S6, enzymolysis：Bovine bone powder after decalcification is handled is poured into the enzyme digestion reaction tank of stainless steel, and toward the deionized water (0.5Kg) that 0.1 times of ox bone weight is added in enzyme digestion reaction tank, and add the pepsin (5g) of ox bone weight 0.1%, then 30 DEG C are warming up to the progress of enzyme digestion reaction tank while stirring, enzymolysis 2 hours, while the pH value in controlled enzymatic hydrolysis retort is 2.5；
S7, go out enzyme：After enzymolysis is finished, enzyme digestion reaction tank is warming up to 100 DEG C, the slurry micro-boiling 5 minutes after enzymolysis is kept, pepsin is lost activity；
S8, filtering：After the enzyme that goes out is finished, enzyme digestion reaction tank is cooled to 50 DEG C, then will be filtered using vertical diatomite filtering machine, and be removed filter residue, obtain collagen solution；
S9, decolouring：Activated carbon is added into collagen solution and carries out decolorization, the addition of 20 mesh particle diameter activated carbon granules is 0.1 times (0.5kg) of collagen solution weight；
S10, concentration：The activated carbon after decolorization is filtered, and the filtrate obtained after filtering is passed through in batch (-type) single-action coiled vacuum concentration pan according to 550kg/h flow, and vacuum concentration processing is carried out at for 65 DEG C, moisture removal is removed, obtains solid matter；
S11, drying：The solid matter that vacuum concentration processing is obtained, which is placed in the drying box of air-dry machine, is dried processing；Wherein, the EAT of air-dry machine is 230 DEG C, and leaving air temp is 95 DEG C, obtains ox bone collagen albumen.
Embodiment 2：
The step of preparation process of ox bone collagen albumen is as follows：
S1, cleaning：The ox bone of 10Kg two one full year of life oxen is put into rinse bath, then toward clear water is added in rinse bath, the bloodstain sticked on ox bone, viscous thing are rinsed repeatedly；Then in injected clear water and ox bone is flooded completely, then carrying out ultrasonic vibration to rinse bath stirs 60 minutes；Repeat the step 4-6 times, it is standby until ox bone surface clean is clean to remove the impurity on ox bone surface；
S2, drying：Clean ox bone is put into drying baker, dried 24 hours under air atmosphere at 60 DEG C；In drying process, an ox bone was stirred every 2 hours, ox bone is gone moisture removal, uniform drying as far as possible；
S3, crushing：After with pulverizer, dry ox bone is crushed again on ball mill ball milling into the bovine bone powder that particle diameter is 200 microns；For ease of crushing, before crushing, ox bone can first be cut into the fine granularity of small dimension, then pulverization process is being carried out with pulverizer；Ball mill is ground using agate ball, and sphere diameter control is at 5 ~ 20 millimeters；
S4, degreasing：Bovine bone powder is placed under 60 DEG C of water bath conditions and carries out low temperature reflux degreasing 5 hours, and reflux temperature control is at 100 DEG C or so；
S5, decalcification：The bovine bone powder of ungrease treatment is put into 10% hydrochloric acid solution and soaked 24 hours, decalcification processing is carried out, and a hydrochloric acid solution was changed every 3 hours, hydrochloric acid solution is cleaned up repeatedly with clear water again；
S6, enzymolysis：Bovine bone powder after decalcification is handled is poured into the enzyme digestion reaction tank of ceramics, and toward the deionized water (5Kg) that 0.5 times of bovine bone powder weight is added in enzyme digestion reaction tank, and add the SUKAPro AC (50g) of bovine bone powder weight 0.5%, then 50 DEG C are warming up to the progress of enzyme digestion reaction tank while stirring, enzymolysis 5 hours, while the pH value in controlled enzymatic hydrolysis retort is 4；
S7, go out enzyme：After enzymolysis is finished, enzyme digestion reaction tank is warming up to 100 DEG C, the slurry micro-boiling 10 minutes after enzymolysis is kept, SUKAPro AC is lost activity；
S8, filtering：After the enzyme that goes out is finished, enzyme digestion reaction tank is cooled to 60 DEG C, then will be filtered using vertical diatomite filtering machine, and be removed filter residue, obtain collagen solution；
S9, decolouring：Activated carbon is added into collagen solution and carries out decolorization, the addition of 40 mesh particle diameter activated carbon granules is 0.1 times (1kg) of collagen solution weight；
S10, concentration：The activated carbon after decolorization is filtered, and the filtrate obtained after filtering is passed through in batch (-type) single-action coiled vacuum concentration pan according to 550kg/h flow, and vacuum concentration processing is carried out at 70 DEG C, moisture removal is removed, obtains solid matter；
S11, drying：The solid matter that vacuum concentration processing is obtained, which is placed in the drying box of air-dry machine, is dried processing；Wherein, the EAT of air-dry machine is 230 DEG C, and leaving air temp is 95 DEG C, obtains ox bone collagen albumen.
Claims (3)
1. a kind of preparation method of ox bone collagen albumen, it is characterised in that comprise the following steps：
S1, cleaning：Half ox bone more than one full year of life, below two one full year of life is put into rinse bath, then toward adding clear water in rinse bath and flooding ox bone completely, ultrasonic agitation then is carried out 30 ~ 60 minutes to rinse bath；The step is repeated repeatedly, to remove the impurity on ox bone surface；
S2, drying：Clean ox bone is dried 12 ~ 24 hours at 60 ~ 80 DEG C；
S3, crushing：After with pulverizer, dry ox bone is crushed again on ball mill ball milling into the bovine bone powder that particle diameter is 5 ~ 200 microns；
S4, degreasing：Bovine bone powder is placed under 60 DEG C of water bath conditions and carries out low temperature reflux degreasing 4 ~ 5 hours；
S5, decalcification：Bovine bone powder after ungrease treatment is put into 10% hydrochloric acid solution and soaked 12 ~ 24 hours, decalcification processing is carried out, and a hydrochloric acid solution was changed every 3 hours, hydrochloric acid solution is cleaned up repeatedly with clear water again；
S6, enzymolysis：Bovine bone powder after decalcification is handled is poured into enzyme digestion reaction tank, and toward the deionized water that 0.1 ~ 0.5 times of bovine bone powder weight is added in enzyme digestion reaction tank, and add the acid protease of bovine bone powder weight 0.1 ~ 0.5%, then 30 ~ 50 DEG C are warming up to the progress of enzyme digestion reaction tank while stirring, enzymolysis 2 ~ 5 hours, while the pH value in controlled enzymatic hydrolysis retort is 2.5 ~ 4；
S7, go out enzyme：After enzymolysis is finished, enzyme digestion reaction tank is warming up to 100 DEG C, the slurry micro-boiling 5 ~ 10 minutes after enzymolysis is kept, acid protease is lost activity；
S8, filtering：After the enzyme that goes out is finished, enzyme digestion reaction tank is cooled to 50 ~ 60 DEG C, then will be filtered using vertical diatomite filtering machine, and be removed filter residue, obtain collagen solution；
S9, decolouring：Activated carbon is added into collagen solution and carries out decolorization, the addition of activated carbon is 0.1 ~ 0.5 times of collagen solution weight；
S10, concentration：The activated carbon after decolorization is filtered, and the filtrate obtained after filtering is passed through in batch (-type) single-action coiled vacuum concentration pan according to 550kg/h flow, and vacuum concentration processing is carried out at for 65 ~ 70 DEG C, moisture removal is removed, obtains solid matter；
S11, drying：The solid matter that vacuum concentration processing is obtained, which is placed in the drying box of air-dry machine, is dried processing；Wherein, the EAT of air-dry machine is 230 DEG C, and leaving air temp is 95 DEG C, obtains ox bone collagen albumen.
2. the preparation method of ox bone collagen albumen according to claim 1, it is characterised in that the acid protease is pepsin, SUKAPro AC, 537 acid proteases.
3. the preparation method of ox bone collagen albumen according to claim 1, it is characterised in that the activated carbon is granular active carbon, and particle diameter is 20 ~ 40 mesh.
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| CN107630061A (en) * | 2017-09-29 | 2018-01-26 | 中国农业科学院农产品加工研究所 | The preparation method of Yak Bone NTx albumen |
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| CN109369800A (en) * | 2018-11-09 | 2019-02-22 | 贵州梵净山生态农业股份有限公司 | The extracting method of high-purity collagen in a kind of ox bone |
| CN109369800B (en) * | 2018-11-09 | 2021-12-21 | 贵州梵净山生态农业股份有限公司 | Method for extracting high-purity collagen from bovine bone |
| CN110564802A (en) * | 2019-09-30 | 2019-12-13 | 南宁学院 | Extraction method of yak achilles tendon bone collagen |
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| CN113201065B (en) * | 2021-04-19 | 2022-05-20 | 国肽生物工程（常德）有限公司 | Bovine bone collagen peptide with functions of relieving fatigue and improving bone density and preparation method thereof |
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