CN101289507A - Collagen protein and collagen polypeptides, preparation thereof and applications - Google Patents
Collagen protein and collagen polypeptides, preparation thereof and applications Download PDFInfo
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Abstract
The invention provides a collagen and a collagen polypeptide, wherein, the weight percentage of hydroxylysine is more than 1.3 percent. The invention also provides the preparation technology and the application of the collagen and the collagen polypeptide. The invention can make full use of inexpensive raw materials including the fish skin and fish scale of tilapias to produce products with high added value. The collagen polypeptide is used for treating arthritis with a definite drug effect, and the clinical dose is considerably less than other collagen polypeptides. The collagen polypeptide can solve the pollution problem of fish skin wastes, and at the same time avoid the risk of infectious diseases which can be caused by the collagen of land animals. The collagen and the collagen polypeptide have the advantages of considerable economic benefits, energy conservation and environmental protection, simple process equipment, low cost and easy industrialization.
Description
Technical field
The present invention relates to a kind of collagen protein and collagen polypeptide, particularly derive from collagen protein and collagen polypeptide and the enzymolysis preparation and the application of fish-skin or fish scale.
Background technology
Collagen protein and collagen polypeptide have been widely used in fields such as medicine, chemical industry, food at present, comprise medical material, medicine, healthcare products, also comprise cosmetics and skincare product, the particularly effect of the hemostatic function of collagen uniqueness, the growth of promotion cell and wound healing, and biocompatibility and biological degradability, make collagen protein very obvious as the advantage of healthy products raw material.This also is a domestic and international research focus in recent years.Therefore make full use of the collagen protein biomass resource, have a extensive future.
The preparation raw material of collagen is mainly from mammiferous bone, skin etc. at present, because pestilence in the animal in recent years, bring very big threat to the mankind, as mad cow disease, foot and mouth disease, bird flu etc., cause all over the world fear to the collagen product of these animal-origins, therefore, seeking animal material beyond the Mammals prepares colory collagen and has become that those skilled in the art institute expects jointly.Like this, the former series products of fish glue just becomes and has been popular.
China's fresh water has the squama fish resource very abundant, is again the producing country and the export State of tilapia maximum, and only outlet every year of fish skin and scale is with regard to few hundred thousand tonnes of.
Isin glue collagen contains 19 seed amino acids (more than 7 seed amino acids and 2 kinds of children's indispensable amino acids of comprising needed by human, the contained amino acid of people lactoprotein two kinds, its protein content is higher 4 times than milk powder, sheep milk powder).
Polypeptide more and more comes into one's own in recent years, modern biology experiment is verified, and the fish skin and scale collagen polypeptide is compared with amino acid with protein has more advantages: physiological functions such as excellent absorption, low antigenicity, promotion lipid metabolism, reducing cholesterol, promotion mineral substance absorb, hypotensive, anti-oxidant; Excellent water-absorbent and water-retentivity.
Usually, the characteristic of collagen and its preparation technology are closely related, existing collagen goods mainly are to adopt gelatin degraded, animal osteoderm High Temperature High Pressure to put forward glue technology such as enzymolysis again, cause active reduction of effect of product, energy consumption is too high, also may mix some objectionable impuritiess or the like, and it is existing from the technology of fish skin and scale processed collagen albumen and polypeptide, what mainly adopt also is that High Temperature High Pressure is put forward the glue technology of enzymolysis again, has destroyed its original constitutional features, causes active reduction.(as: Fu Yanfeng, Shen Yuexin, brief talk the utilization of collagen of fish skin, food research and development, the 25th the 2nd phase of volume of April in 2004) the tall and erect equality of fourth, the research of tilapia fishskin collagen protein extraction conditions, aquatic science and technology information, 2006,33 (4) have reported extracted collagen from Java tilapia skins, concrete grammar is: add different concns, different sorts, the swelling of different time is carried out in the acid of different volumes, homogeneous, vacuum filtration obtains the extract of collagen protein, measure the content of noncollagen protein, hydroxyproline content (purity testing), the content of oxyproline are 4.35%.Oxyproline is a kind of specific proteins of collagen protein, and is normal at present by Determination of Hydroxyproline being determined the content of collagen protein in the tilapia fishskin.But, because the steady chemical structure of oxyproline is very high, in the collagen protein of selling in the market, no matter adopt which kind of technology, the content of oxyproline is very little, its content is relevant with animal-origin in other words, and have no relations with technology, also little with the relation of effect quality, therefore, adopting oxyproline is the quality of index reflection collagen protein, has little significance.
Document also has report to extract collagen protein from fish scale, as: Liu Qinghui etc., fish scale collagen research, marine fishery research, September in 2000, the 21st the 3rd phase of volume adopted acidity and enzyme extraction method to extract solubility in acid and enzymatic dissolubility type i collagen albumen from fish scale, and wherein fish scale is after fetch the fish market, through pre-treatment, adopt 1% protease treatment, dialysis is purified, wherein, the percentage composition of hydroxylysine is 0.60%, 0.70%, and the percentage composition of oxyproline is 6.65%, 6.08%.Wang Nanping, Guo Pengda, the development of fish scale collagen, aquatic science and technology information, 2004,31 (6) have reported the collagen protein manufacture craft, with fish scale cleaning, degreasing, deliming, hot melt process, enzymolysis, decolouring, smart filter, concentrate, sterilization, drying, the collagen protein powder.Wherein, the percentage composition of hydroxylysine is 0.80%, and the percentage composition of oxyproline is 10.1%.At the molecular weight of collagen protein, also mention in the document: " must detect its molecular weight size with electrophoretic method as the collagen protein product, generally with 5000-20000 for well, and dissolubility test, pH is neutral, the 400nm transmitance is more than 90%." hydroxylysine exists only in the collagen protein, is the glycosylation site of collagen in biochemical cycle, is the active in vivo important structure of collagen polypeptide site.Except above-mentioned non-patent literature report, the report that also has patent documentation that fish scale collagen is correlated with, as: application number: 200310114500.X, denomination of invention: the production method of fish scale collagen, the present invention relates to a kind of production method of fish scale collagen, it selects for use the fine fish scale to make raw material, carry out pre-treatment, remove thick substances and other impurity on fish scale surface, and after drying removes raw meat, then adopt acid-base method to carry out deliming, degreasing, deliming, the laggard horizontal high voltage catalysis of degreasing is steamed and is separated, and refines liquid low molecular protein peptide fish scale collagen product.The present invention has made full use of aquatic resources, and the fish scale waste is turned waste into wealth, and has both reduced the pollution to environment, has fundamentally guaranteed the quality of collagen protein again; Production technique is simple, strong operability, and cost is low.Do raw material with it, can various nutritive mediums of processing and fabricating and protective foods, be beneficial to very much absorption of human body.Application number: 200510126543.9, denomination of invention: utilize enzyme engineering technology from fish scale, to extract the method for micromolecular collagen, the present invention utilizes enzyme engineering technology to extract the method for micromolecular collagen from fish scale, comprise and adopt fish scale, and utilize the biological enzyme incision technology from fish scale, to extract the small molecules fish scale collagen as main raw material of the present invention; Producing of described small molecules fish scale collagen, must through the screening of fish scale is cleaned, is dried, crushing screening, water distribution is stirred, and utilizes protease hydrolyzed, the enzyme that goes out, filtration.After the concentrating filter liquor that obtains, drying, promptly obtain the small molecules fish scale collagen.Application number: 200510044916.8, denomination of invention: a kind of Fishscale collagen production process, the invention discloses a kind of Fishscale collagen production process, be through twice acid treatment of fish scale, the alkali neutralization, boil, enzymolysis, exchange absorption, concentrate, filter, dry spraying obtains, the invention provides a kind of industrial waste fish scale that made full use of turns waste into wealth, it is low and stable that the Fishscale collagen production process of the simple and safe easy handling of processing step, the product of production have a molecular weight, product does not have the fishy smell bitter taste, the ash content of coal is extremely low, the production efficiency height, fish scale is the characteristics of enzymolysis thoroughly.
Glucosamine glucosamine is that a hydroxyl of glucose is by an amino-substituted compounds.Molecular formula C
6H
13O
5N is commonly called as aminosugar.Extensively be present in nature, 2-amino-2-deoxy-D-glucose is present in the polysaccharide of microorganism, animal-origin and in conjunction with in the polysaccharide with N-acetyl derivative (as chitin) or with N-sulfuric ester and N-acetyl-3-O-lactic acid ether (teichoic acid) form usually.Glucosamine is a kind of derivative of natural amino monose, is the necessary important component of synthetic proteins glycan in the cartilage matrix.Proteoglycan can make joint cartilage have the function that absorbs surging force by the drawing force that suppresses collegen filament.As: Liu Nian, glucosamine, chondroitin sulfate and two medicines unite and are used for the knee osteoarthritis treatment of pain, " foreign medical science: pharmacy fascicle "-2006 33 volumes 4 phases-254-254 page or leaf.But, glucosamine and arthritic compatibility of drugs are used, can bring into play synergistic function, for a person skilled in the art, be unpredictable.
Summary of the invention
Technical scheme of the present invention has provided a kind of new collagen protein and collagen polypeptide, another technical scheme of the present invention provided this collagen protein and collagen polypeptide purposes and with the pharmaceutical composition of other medicines combined utilization.
The invention provides a kind of collagen protein and collagen polypeptide, wherein, the hydroxylysine weight percentage is greater than 1.3%; Its weight-average molecular weight is 2000～4000.
The present invention also provides a kind of technology for preparing collagen protein and collagen polypeptide, may further comprise the steps:
A, with the dried animal material that can be used for preparing collagen of 100 weight parts, add 800～1200 weight parts waters, be warming up to 80～90 ℃, constant temperature 20～60 minutes;
B, be cooled to 30～65 ℃ then, regulate pH value to 8～9, add 1～8 part of Sumizyme MP, constant temperature was degraded 0.5～12 hour;
C, again under 45-50 ℃ of constant temperature, add 1～8 part of neutral protease, stirring reaction 0.5～12 hour;
D and then be warmed up to 75～90 ℃ is incubated 15～30 minutes, makes enzyme deactivation;
E, adding flocculation agent through flocculation sediment impurity, filtration, in vacuum spray drying, promptly get solid state collagen albumen and collagen polypeptide powder with filtrate.
Further preferably,
A, with the dried animal material that can be used for preparing collagen of 100 weight parts, add 800～1200 weight parts waters, be warming up to 80～85 ℃, constant temperature 30 minutes;
B, be cooled to 45～50 ℃ then, regulate pH value to 8, add 2.5～5 parts of Sumizyme MPs, constant temperature was degraded 2～6 hours;
C, again under 45 ℃ of constant temperatures, add 3～5 parts of neutral proteases, stirring reaction 2～5 hours;
D and then be warmed up to 85～90 ℃ is incubated 15～30 minutes, makes enzyme deactivation;
E, adding flocculation agent, flocculation agent is the mixture of diatomite and gac, weight proportion is 1: (1-2) mix, through flocculation sediment impurity, filtration, filtrate in vacuum spray drying, is promptly got solid state collagen albumen and collagen polypeptide powder.
Preferably, the described animal material of step a is fish-skin or fish scale; The described Sumizyme MP of step b is at least a in alcalase or the pancreatin; The described neutral protease of step c is A1398 or Neutrase enzyme, or at least a among the Flavourzyme; The described flocculation agent of step e is at least a of diatom great soil group or gac.
The present invention also provides described collagen protein or the application of collagen polypeptide in the preparation skin-care product.
The present invention also provides collagen protein or the application of collagen polypeptide in the preparation treatment of arthritis.
The present invention also provides a kind of combination medicine of treatment of arthritis, and it comprises the collagen protein and collagen polypeptide and the coupling glucosamine that are used for while, difference or administration successively in the different preparation units, and pharmaceutically useful carrier.
Wherein, the dosage of described collagen protein and collagen polypeptide and glucosamine is singly used effective dose for being lower than.
The common using dosage of present commercially available collagen polypeptide is 10g/ time, (sees: Role ofCollagen Hydrolysate in Bone and Joint Disease Semin Arthritis Rheum.2000Oct for 1 time every day; 30 (2): 87-99).
The present invention uses the dosage of collagen protein to be separately 5g/ time, every day 1 time, can use in clinical to be lower than 3g/ time; Unite use with glucosamine, the collagen protein using dosage is 3g/ time, every day 1 time; Use with collagen protein of the present invention and collagen polypeptide compatibility, the using dosage of glucosamine is: 600mg/ day, (the conventional using dosage of glucosamine was: 1000mg/ day).
Wherein, the weight proportion of collagen protein and collagen polypeptide and glucosamine is:
Collagen protein and collagen polypeptide 70-95 part, glucosamine 5-30 part.
Further preferentially, the weight proportion of collagen protein and collagen polypeptide and glucosamine is
Collagen protein and collagen polypeptide 70-80 part, glucosamine 20-30 part.
Still more preferably, the weight proportion of collagen protein and collagen polypeptide and glucosamine is:
80 parts of collagen protein and collagen polypeptides, 20 parts of glucosamines.
Preparation collagen protein and polypeptide, prior art generally comprises: degreasing, take off foreign protein, liming, acid neutralization, high temperature and put forward glue, normal temperature enzymolysis, deactivation, spraying drying.Compare with existing disclosed technology, in the collagen polypeptide preparation process, the present invention has omitted the treatment process in early stage, directly skips gelatin extraction technique, under the extraction preparation condition of gentleness, with animal material, particularly fish skin and scale is by enzymolysis, the flocculation sediment removal of impurity (comprising foreign protein and pigment), concentrated, dry, final collagen protein enzymolysis and polypeptide powder, carry glue at a lower temperature and guaranteed the biological activity of collagen.
Because the present invention in the preparation process of collagen polypeptide, has simplified earlier stage treatment process, not with an organic solvent, do not use strong oxidizer decolouring, deodorization, do not produce secondary pollution, the finished product do not contain harmful impurity; Simultaneously, the molecular weight size and the distribution of may command bioactive peptide, make it to be stabilized in 2000～4000, the molecular weight homogeneous, the collagen polypeptide of this molecular weight ranges is very easy to be absorbed, and is widely used in the cosmetics, is used for the treatment of sacroiliitis, drug effect is clear and definite, and clinical using dosage is starkly lower than other collagen polypeptide; The water retention property excellence, compared with similar products, under the equivalent weight condition, can be 25 ℃ of temperature, 80% ambient moisture absorbed the airborne moisture of 1～2 times of weight part in 3 hours.
In addition, the present invention can make full use of inexpensive raw material Java tilapia skin fish scale, the product that the production added value is high, can solve the pollution problem of fish-skin waste again, avoid the risk of the transmissible disease that the terricole collagen protein may bring simultaneously, economic benefit is considerable, and energy-conserving and environment-protective, processing unit is simple, and is with low cost, is easy to industrialization.
Description of drawings
Fig. 1 is the molecular weight determination collection of illustrative plates (wherein, molecular weight is 3900 dalton, uv-absorbing wavelength 215 nanometers) of the collagen protein of embodiment 1 preparation;
Fig. 2 is the molecular weight determination collection of illustrative plates (wherein, molecular weight is 3400 dalton, uv-absorbing wavelength 215 nanometers) of the collagen protein of embodiment 2 preparations.
Embodiment
Technical process:
Dried fish-skin (or frozen fish skin), decalcification fish scale → cleaning → enzymolysis → inactivator → flocculation sediment → filtration → spraying drying (or freeze-drying) → sterilization
Embodiment 1 is feedstock production collagen protein and polypeptide with the tilapia squama
With 100 gram decalcification tilapia squamas, washing, add deionized water 1000 be warming up to 85 ℃ 30 minutes, after be cooled to 45 ℃, adjust pH to 8.0 adds 3 gram Sumizyme MPs, degraded 2 hours, and added neutral protease 4 grams under the similarity condition again, reacted 2 hours, scale all dissolves, be warmed up to 90 ℃ then, be incubated 15～30 minutes, make enzyme deactivation, add an amount of flocculation agent diatomite and gac (mixing) sedimentation 8 hours, directly get supernatant with 1: 1; Supernatant liquor is in vacuum tightness 0.06～0.10Mpa, 40～65 ℃ of underpressure distillation of temperature, and last spraying drying promptly gets the solid-state protein polypeptide powder.
After measured, wherein: hydroxyproline content: 9.4%; Hydroxylysine, 1.4%.
Embodiment 2 is feedstock production collagen protein and polypeptide with the Java tilapia skin
Take by weighing the dried fish-skin of 100g in the 1000ml beaker, normal temperature wash to the pH value be about 7, be heated to down constant temperature 30 minutes of 80 degree, be cooled to 50 ℃, adjust pH to 8 adds Sumizyme MP 2.5 grams, reacts 3 hours; Temperature adjustment to 45 ℃ adds neutral protease 4 grams, constant temperature stirring reaction 3 hours; Be heated to 85 ℃ of constant temperature 15 minutes, inactivator.Add an amount of flocculation agent diatomite and gac (mixing) sedimentation 8 hours, directly get supernatant with 1: 2.Supernatant liquor is in vacuum tightness 0.06～0.10Mpa, 40～65 ℃ of underpressure distillation of temperature, and last spraying drying promptly gets the solid-state protein polypeptide powder.
Product detected result: hydroxyproline content: 9.38%; Hydroxylysine, 1.31%.
Embodiment 3 is feedstock production collagen protein and polypeptide with the cod skin
Take by weighing the dry cod skin of 100g in the 1000ml beaker, normal temperature wash to pH value be about 7, be heated to 80 degree constant temperature 30 minutes down, be cooled to 50 ℃, adjust pH to 8, pancreatin 4 restrains in adding respectively, reacts 6 hours; Temperature adjustment to 45 ℃ adds 1398 protease 3s gram, constant temperature stirring reaction 5 hours; Be heated to 85 ℃ of constant temperature 15 minutes, inactivator.
Filter through flocculation sediment, filtrate is carried out underpressure distillation in about 50 ℃, spray-dried at last, promptly gets the collagen of fish skin polypeptide powder.
Product detected result: hydroxyproline content: 7.3%; Hydroxylysine, 1.3%
Embodiment 4 is feedstock production collagen protein and polypeptide with the tilapia squama
Process is with embodiment 1, and wherein, Sumizyme MP is a pancreatin, consumption 8 gram, and neutral protease 8 gram flocculation agents are diatomite, supernatant liquor is in vacuum tightness 0.06～0.10Mpa, 40～65 ℃ of underpressure distillation of temperature, last spraying drying promptly gets the solid-state protein polypeptide powder.
After measured, wherein: hydroxyproline content: 9.2%; Hydroxylysine, 1.42%
Embodiment 5 is feedstock production collagen protein and polypeptide with the grass carp squama
With 100 gram decalcification grass carp squamas, washing adds deionized water 1000 and was warming up to 85 degree 30 minutes, after be cooled to 45 ℃, adjust pH to 8.0 adds 5 gram alcalase enzymes, degraded 2 hours, and added 1398 enzymes, 3 grams under the similarity condition again, reacted 2 hours, scale all dissolves, be warmed up to 90 ℃ then, be incubated 15～30 minutes, make enzyme deactivation, add mixture (mixing) sedimentation 8 hours of an amount of flocculation agent diatomite and gac, directly get supernatant according to 1: 2.Supernatant liquor is in vacuum tightness 0.06～0.10Mpa, 40～65 ℃ of underpressure distillation of temperature, and last spraying drying promptly gets the solid-state protein polypeptide powder.
After measured, wherein: hydroxyproline content: 9.8%; Hydroxylysine, 1.43%.
Test example 1 collagen protein and collagen polypeptide property testing
During collagen protein and collagen polypeptide are soluble in water, place the easy moisture absorption in the air.Compared with similar products, under the equivalent weight condition, can be 25 ℃ of temperature, 80% ambient moisture absorbed the airborne moisture of 1～2 times of weight fraction in 3 hours.Contrast and experiment sees the following form:
The chromatograph (AKTA Explorer 100) that adopts Pharmacia (Pharmacia) company to produce is analyzed, and molecular-weight average about 2000～4000, average peptide chain length are 100～130 and 10～13.
Amino acid analysis (being the detected result of embodiment 1 product):
| Asp, the % asparagus fern | / | 5.10 |
| Thr, % Su An | / | 2.71 |
| Ser, % silk peace | / | 3.26 |
| Glu, % paddy peace | / | 9.38 |
| Gly, the sweet peace of % | / | 20.55 |
| Ala, % third peace | / | 8.83 |
| Cys, % Guang peace | / | 0.18 |
| Val, % figured silk fabrics peace | / | 2.30 |
| Met, % egg peace | / | 1.44 |
| Ile, the different bright peace of % | / | 1.45 |
| Leu, the bright peace of % | / | 2.75 |
| Tyr, % junket peace | / | 0.81 |
| Phe, the % phenylpropyl alcohol | / | 2.09 |
| Lys, % Methionin | / | 3.01 |
| Holys % hydroxylysine | / | 1.43 |
| Ammonia, % ammonia | / | 0.62 |
| His, % organizes peace | / | 1.12 |
| Arg, the smart peace of % | / | 7.08 |
| Pro, % dried meat peace | / | 12.98 |
| HOPro % hydroxyl dried meat peace | / | 9.41 |
| Try, % look peace | / | / |
| Total amino acids，％ | ≥70 | 95.89 |
The comparison of test example 2 collagens of the present invention and collagen polypeptide and currently available products
The collagen polypeptide performance comparison sheet of different process and animal-origin
| Fish-skin (embodiment 2) | Fish scale (embodiment 1) | Pigskin | Gelatin hydrolysate | |
| Water-soluble (25 ℃ are not less than 10g/100ml water) | Molten entirely | Molten entirely | Molten entirely | Molten entirely |
| PH value (1% the aqueous solution) | 6 | 6 | 5.5 | 5.5 |
| Moisture, %≤ | 5 | 4 | 7 | 7 |
| Hydroxylysine content, % 〉= | 1.32 | 1.41 | 1.2 | 1.1 |
| Aqueous solution color and luster (10% concentration) | Faint yellow transparent | Faint yellow transparent | Faint yellow transparent | Faint yellow transparent |
| The repairing performance (was life cycle with 24 hours) that is used for trauma repair essence (content 10%) | Well | Well | Well, but taste be difficult to accept | Do not appear in the newspapers |
| Be used for after the nutritional supplementation special efficacy (with continuous oral 5 days, every days 5 gram, the sensation of skin after 5 days, number of users 10 people) | Well | Obviously improve | Well | No change almost |
| Cost | Low | Low | Higher | High |
| The production environment influence | Well | Well | Pollution is arranged | Pollution is arranged |
| Total nitrogen, % 〉= | 13 | 14 | 10 | 10 |
| Arsenic (in As), mg/kg≤ | 0.5 | 0.5 | 0.5 | 0.5 |
| Plumbous (in Pb), mg/kg≤ | 1 | 0.5 | 1 | 1 |
| Molecular weight and concentration degree with AKATA Explorer 100 chromatographs, detect | 2000～7000, unimodal, 98% | 2000-4000, unimodal, 99% | 2000～7000, three peaks, 97% | 2000～7000, a plurality of peaks, 86-90% |
" well " described in the table refers to the makeup feel exquisiteness for preparing, and effect is reliable, does not have other side effect; " better " refers to have certain effect, but do not reach the standard of " well ".
Above-mentioned evidence, technology of the present invention is easy, low production cost, prepared collagen and collagen polypeptide molecular weight homogeneous, performance has remarkable distinctive characteristics and significant superiority than currently available products, and fabulous prospects for commercial application is arranged.
The application of test example 3 scale collagen polypeptides in arthritis treatment
Material and method:
Case is selected: (1) adult man or women, age 45-70a; (2) meet the patient that osteoarthritis is diagnosed; (3) arthrodynia and the dysfunction that has osteoarthritis such as knee, hip to cause; (5) scoring of the visual simulation scale (VAS) of level walking pain is 4～6 minutes (comprising 4 fens and 6 minutes);
Methods of treatment:
1, the test scale collagen polypeptide powder of the method preparation of product: embodiment 1, the glucosamine hydrochloride that Haipu Bio-Technology Co Ltd, Qingdao buys, room temperature preservation.Scale collagen polypeptide powder: 3 gram/bags; Ammonia sugar collagen peptide powder, 3 gram/bags wherein contain 600 milligrams of ammonia sugar.(wherein the weight compatibility of glucosamine and collagen polypeptide is: 20: 80.)
2, the instructions of taking and the course of treatment: nutrient drug adopts the random device grouping, test group (pure powder group): oral, and each 1 bag, every day 1 time.Test group (ammonia sugar collagen peptide group): oral, each 1 bag, every day 1 time.Taking 4wk continuously is a course of treatment.Be a course of treatment observing time.
Efficacy determination:
1, main curative effect index
Main curative effect index quantizes grade form as main efficacy evaluation instrument for the osteoarthritis state of an illness that adopts design voluntarily.The parameters for observation on effect total points is according to playing the ankylosis scoring gained that adds up in articular pain, level walking pain, rest pain, morning.Being assessed as according to judgement criteria: clinical complete remission---the improvement rate is more than or equal to 90%; Produce effects---the improvement rate is more than or equal to 60%, less than 90%; Effectively---the improvement rate is more than or equal to 30%, less than 60%; Invalid---the improvement rate is less than 30%.Total points * 100% before improvement rate=(total points one treatment back total points before the treatment)/treatment.
Efficient=(alleviating case load+produce effects case load+effective case load fully)/total case load * 100%.
The result:
In December, 2006～2007 amount to case load 284 examples that reception is taken year December, are divided into pure scale collagen polypeptide group and ammonia sugar collagen peptide group, and every group is respectively 133 example and 151 examples.The male sex's 112 examples wherein, women's 172 examples.Be distributed in Beijing, Dandong City, Jilin, Wuhan City, Chengdu, Sichuan Province.
Treatment situation improvement rate therapeutic evaluation comparative example (%)
Simultaneously, collect user's information of some commercially available collagen proteins, comparing result is:
Above-mentioned evidence, collagen polypeptide treatment of arthritis of the present invention, drug effect is obvious, obviously is better than commercially available collagen protein, and compares with commercially available collagen protein, effective using dosage low (commercially available collagen protein effective usage consumption is 10g/ days), drug effect height.
When uniting use with glucosamine, use more to be lower than the routine clinical using dosage, just reached obvious effects in first cycle (in 7 days), be better than using collagen protein of the present invention separately, have the effect of synergy.
The compatibility shaker test of test example 4 glucosamines and collagen polypeptide
Operate by test example 3 described methods, glucosamine and collagen polypeptide shaker test the results are shown in following table:
By above-mentioned test-results explanation, glucosamine and collagen polypeptide compatibility use, weight proportion is: glucosamine: collagen polypeptide (5-30): (70-95) all can reach the effect of synergy preferably, wherein more excellent weight compatibility scope is: glucosamine: collagen polypeptide (20-30): (70-80).
Above-mentioned evidence, the hydroxylysine weight percentage no matter from physical and chemical index, or drug effect, all was better than existing collagen protein and collagen polypeptide products greater than 1.3% o'clock in collagen protein and the collagen polypeptide.As skin-care product, has good effect; Be used for the treatment of sacroiliitis, obviously be better than commercially available collagen protein, and compare with commercially available collagen protein, effectively using dosage is low; When uniting use with glucosamine, use more to be lower than the routine clinical using dosage, have synergistic function, economic benefit is considerable, and energy-conserving and environment-protective, and processing unit is simple, and is with low cost, is easy to industrialization.
Claims (11)
1, a kind of collagen protein and collagen polypeptide is characterized in that: the hydroxylysine weight percentage is greater than 1.3%.
2, collagen protein according to claim 1 and collagen polypeptide is characterized in that: weight-average molecular weight is 2000～4000.
3, a kind of technology for preparing claim 1 or 2 described collagen proteins and collagen polypeptide may further comprise the steps:
A, with the dried animal material that can be used for preparing collagen of 100 weight parts, add 800～1200 weight parts waters, be warming up to 80～90 ℃, constant temperature 20～60 minutes;
B, be cooled to 30～65 ℃ then, regulate pH value to 8～9, add 1～8 part of Sumizyme MP, constant temperature was degraded 0.5～12 hour;
C, again under 45-50 ℃ of constant temperature, add 1～8 part of neutral protease, stirring reaction 0.5～12 hour;
D and then be warmed up to 75～90 ℃ is incubated 15～30 minutes, makes enzyme deactivation;
E, adding flocculation agent through flocculation sediment impurity, filtration, in vacuum spray drying, promptly get solid state collagen albumen and collagen polypeptide powder with filtrate.
4, technology according to claim 3 is characterized in that: the described animal material of step a is fish-skin or fish scale.
5, technology according to claim 3 is characterized in that: the described Sumizyme MP of step b is at least a in alcalase or the pancreatin.
6, technology according to claim 3 is characterized in that: the described neutral protease of step c is A1398 or Neutrase enzyme, or at least a among the Flavourzyme.
7, technology according to claim 3 is characterized in that: the described flocculation agent of step e is at least a of diatomite or gac.
8, claim 1 or 2 described collagen proteins or the collagen polypeptide application in the preparation skin-care product.
9, claim 1 or 2 described collagen proteins or the collagen polypeptide application in the preparation treatment of arthritis.
10, a kind of combination medicine of treatment of arthritis, it comprises the claim 1 or 2 described collagen proteins and collagen polypeptide and the coupling glucosamine that are used for while, difference or administration successively in the different preparation units, and pharmaceutically useful carrier.
11, the combination medicine of treatment of arthritis according to claim 10 is characterized in that: the dosage of described collagen protein and collagen polypeptide and glucosamine is singly used effective dose for being lower than.
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| WO2010043073A1 (en) * | 2008-10-16 | 2010-04-22 | Shi Zongjie | Collagen polypeptide, preparation method and uses thereof |
| CN101899491A (en) * | 2010-05-18 | 2010-12-01 | 武汉艾诗妮娜生物科技有限公司 | Method for preparing micromolecular collagen peptide from fish scales by utilizing complex enzyme |
| CN102676619A (en) * | 2011-12-20 | 2012-09-19 | 浙江省海洋开发研究院 | Comprehensive utilization process for marine fish skins |
| CN103082080A (en) * | 2013-02-07 | 2013-05-08 | 广东海洋大学 | Fish skin and fish scale protein enzymolysis powder with function of relieving fatigue |
| CN103211147A (en) * | 2013-04-08 | 2013-07-24 | 湖北汇丰医药科技有限公司 | Collagen nutrition powder for improving skin activity and whitening skin and preparation method thereof |
| CN103609935A (en) * | 2013-11-13 | 2014-03-05 | 青岛贝尔特生物科技有限公司 | Dietary nutritious supplementary for preventing osteoarticular diseases of middle aged and elderly people |
| CN103893822A (en) * | 2014-04-07 | 2014-07-02 | 四川师范大学 | Collagen polypeptide burn repair film and preparation method thereof |
| CN104223115A (en) * | 2013-06-17 | 2014-12-24 | 史宗洁 | Novel application of fish scale collagen protein |
| CN104351457A (en) * | 2014-11-19 | 2015-02-18 | 乳山市华隆生物科技有限公司 | Fish scale oligopeptide powder and preparation method thereof |
| CN105341767A (en) * | 2015-12-09 | 2016-02-24 | 哈尔滨神守生物科技有限公司 | Preparing method for soft-shelled turtle protein peptide |
| CN106350560A (en) * | 2016-11-08 | 2017-01-25 | 广东海洋大学 | Preparation method of fish protein peptide, prepared fish protein peptide and application |
| CN107073076A (en) * | 2014-11-11 | 2017-08-18 | 株式会社日皮 | Immune activation agent, cellular immunity activator and T cell multiplication agent |
| CN107653291A (en) * | 2017-11-15 | 2018-02-02 | 西藏央金生态农牧科技有限公司 | The standby method for hiding Yak-skin Gelatin original albumen and collagen polypeptide of multi-step enzyme method coordinate system |
| CN108409854A (en) * | 2018-02-10 | 2018-08-17 | 海盐县凌特生物科技有限公司 | A kind of small-molecular-weight silkworm chrysalis collagen |
| CN109350731A (en) * | 2018-12-18 | 2019-02-19 | 汕尾市维明生物科技股份有限公司 | A kind of Rofe fish protein peptide is preparing the application in immune formulation |
| CN113105541A (en) * | 2021-04-23 | 2021-07-13 | 浙江崇山生物制品有限公司 | Method for purifying collagen |
| CN113683678A (en) * | 2021-09-15 | 2021-11-23 | 山西锦波生物医药股份有限公司 | Recombinant I-type humanized collagen C1L2T, and preparation method and application thereof |
| CN114158725A (en) * | 2020-09-10 | 2022-03-11 | 成都恒福禧生物技术有限公司 | Preparation method of ginseng zymolyte containing polypeptide polysaccharide |
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| WO2010043073A1 (en) * | 2008-10-16 | 2010-04-22 | Shi Zongjie | Collagen polypeptide, preparation method and uses thereof |
| CN101899491A (en) * | 2010-05-18 | 2010-12-01 | 武汉艾诗妮娜生物科技有限公司 | Method for preparing micromolecular collagen peptide from fish scales by utilizing complex enzyme |
| CN102676619A (en) * | 2011-12-20 | 2012-09-19 | 浙江省海洋开发研究院 | Comprehensive utilization process for marine fish skins |
| CN102676619B (en) * | 2011-12-20 | 2013-12-25 | 浙江省海洋开发研究院 | Comprehensive utilization process for marine fish skins |
| CN103082080A (en) * | 2013-02-07 | 2013-05-08 | 广东海洋大学 | Fish skin and fish scale protein enzymolysis powder with function of relieving fatigue |
| CN103211147B (en) * | 2013-04-08 | 2015-08-26 | 湖北汇丰医药科技有限公司 | A kind of collagen nutrition powder improving skin vitality skin whitening |
| CN103211147A (en) * | 2013-04-08 | 2013-07-24 | 湖北汇丰医药科技有限公司 | Collagen nutrition powder for improving skin activity and whitening skin and preparation method thereof |
| CN104223115B (en) * | 2013-06-17 | 2017-04-05 | 史宗洁 | The new application of scale collagen polypeptide |
| CN104223115A (en) * | 2013-06-17 | 2014-12-24 | 史宗洁 | Novel application of fish scale collagen protein |
| CN103609935B (en) * | 2013-11-13 | 2016-01-20 | 青岛贝尔特生物科技有限公司 | A kind of dietary supplement preventing middle-aged and old bone and joint diseases |
| CN103609935A (en) * | 2013-11-13 | 2014-03-05 | 青岛贝尔特生物科技有限公司 | Dietary nutritious supplementary for preventing osteoarticular diseases of middle aged and elderly people |
| CN103893822A (en) * | 2014-04-07 | 2014-07-02 | 四川师范大学 | Collagen polypeptide burn repair film and preparation method thereof |
| CN103893822B (en) * | 2014-04-07 | 2015-05-20 | 四川师范大学 | Collagen polypeptide burn repair film and preparation method thereof |
| CN107073076A (en) * | 2014-11-11 | 2017-08-18 | 株式会社日皮 | Immune activation agent, cellular immunity activator and T cell multiplication agent |
| CN104351457A (en) * | 2014-11-19 | 2015-02-18 | 乳山市华隆生物科技有限公司 | Fish scale oligopeptide powder and preparation method thereof |
| CN105341767A (en) * | 2015-12-09 | 2016-02-24 | 哈尔滨神守生物科技有限公司 | Preparing method for soft-shelled turtle protein peptide |
| CN106350560B (en) * | 2016-11-08 | 2020-07-03 | 广东海洋大学 | Preparation method of fish protein peptide, fish protein peptide obtained by preparation method and application of fish protein peptide |
| CN106350560A (en) * | 2016-11-08 | 2017-01-25 | 广东海洋大学 | Preparation method of fish protein peptide, prepared fish protein peptide and application |
| CN107653291A (en) * | 2017-11-15 | 2018-02-02 | 西藏央金生态农牧科技有限公司 | The standby method for hiding Yak-skin Gelatin original albumen and collagen polypeptide of multi-step enzyme method coordinate system |
| CN108409854A (en) * | 2018-02-10 | 2018-08-17 | 海盐县凌特生物科技有限公司 | A kind of small-molecular-weight silkworm chrysalis collagen |
| CN109350731A (en) * | 2018-12-18 | 2019-02-19 | 汕尾市维明生物科技股份有限公司 | A kind of Rofe fish protein peptide is preparing the application in immune formulation |
| CN114158725A (en) * | 2020-09-10 | 2022-03-11 | 成都恒福禧生物技术有限公司 | Preparation method of ginseng zymolyte containing polypeptide polysaccharide |
| CN113105541A (en) * | 2021-04-23 | 2021-07-13 | 浙江崇山生物制品有限公司 | Method for purifying collagen |
| CN113105541B (en) * | 2021-04-23 | 2022-03-11 | 浙江崇山生物制品有限公司 | Method for purifying collagen |
| CN113683678A (en) * | 2021-09-15 | 2021-11-23 | 山西锦波生物医药股份有限公司 | Recombinant I-type humanized collagen C1L2T, and preparation method and application thereof |
| CN113683678B (en) * | 2021-09-15 | 2023-11-24 | 山西锦波生物医药股份有限公司 | Recombinant I-type humanized collagen C1L2T and preparation method and application thereof |
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