CN105219831A - A kind of yak small-molecular peptides of extracting from yak bone and extracting method thereof - Google Patents
A kind of yak small-molecular peptides of extracting from yak bone and extracting method thereof Download PDFInfo
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Abstract
The present invention relates to a kind of yak small-molecular peptides of extracting from yak bone and extracting method thereof, in turn include the following steps: (1) prepares starter; (2) yak bone pre-treatment; (3) elementary fermentable; (4) the elementary yak albumen of electrolysis; (5) alcohol activates the preparation of pancreas slurry; (6) enzymolysis process; (7) last handling process; The molecular weight of described yak small-molecular peptides is that the daltonian accounting of 0-480 is not less than 87.5%; Described yak small-molecular peptides is applicable to diagnosis peptide, peptide vaccine, antibacterial active peptide, cytokine simulating peptide, antiviral peptide or natineoplaston, and peptide health product.
Description
Technical field
The present invention relates to polypeptide preparation technical field, particularly relate to a kind of yak small-molecular peptides of extracting from yak bone and extracting method thereof.
Background technology
Amino acid whose molecule is little, and protein is large, and two or more amino acid dehydrating condensations forms several peptide bonds thus a composition peptide, and multiple peptide carries out a multistage folding just composition protein molecule.Two victory peptides, being called for short dipeptides, is exactly the protein fragments be made up of two amino acid.Having found hundreds of kind peptide in organism, is the requisite participant of physiologically active that body completes various complexity.All cells can improvement on synthesis material, and its functional activity is also by the adjustment of polypeptide.It relates to each field of hormone, nerve, Growth of Cells and reproduction, and its importance is each system organ and cell in control agent.The physiology of enzyme process polypeptide and pharmacological action mainly swash in vivo Enzymes, promote the permeability of intermediary metabolism film, or are transcribed by control DNA or translated and affect special albumen synthesis, finally produce specific physiological effect or play its pharmacological action.Peptide is better than amino acid, and one is that comparatively Amino Acid Absorption is quick; Two is absorbed by body with complete form; Three is active absorption (amino acid belongs to Passive intake); Four is low consumptions, compares with amino acid, and peptide absorbs and has low consumption or do not need catabiotic feature, and peptide, by after duodenal absorption, directly enters blood circulation, by self-energy nutrient delivery to each position of human body; Five is that peptide absorbs comparatively amino acid, has undersaturated feature; Six is that amino acid only has 20 kinds, and function is denumerable, and peptide take amino acid as substrate, can synthesize thousands of kinds up to a hundred.
In many polypeptide, because small active peptides has easy absorption, do not increase human consumption's burden, nutritive value extremely high and extensively realize by contemporary people and accept, become the product of people's health instantly gradually and be subject to heat hold in both hands.The method preparing small-molecular peptides so in the prior art has multiple, such as CN101194733A, CN101532044A, CN102090680A, and CN1280834A all discloses the preparation method about polypeptide, but method is single, extract performance not high, again such as, CN101120724A discloses a kind of microorganism ferment making feed polypeptide formulations, is made up of following weight percent raw material: subtilis 40-55, plant lactobacillus 25-35, lactobacterium casei 10-35.But the polypeptide formulations of preparation is when extracting polypeptide for bone, and extraction efficiency is not high, and bio-fermentation agent is single, and use occasion is also restricted.CN102488073A discloses a kind of extracting method of sea cucumber peptide, adopts enzyme solution, adds sea cucumber specific enzyme, enzymolysis 6 hours, stirs 2 minutes in enzymolysis process every 1 hour; Add gac to de-taste, then hydro-oxidation sodium regulates pH, and measure in concentration process and control degree Beaume 10, the method can not extract more multicomponent polypeptide, and degree Beaume is lower, causes the wasting of resources to a certain degree.CN1931020A discloses a kind of production process of polypeptide of low saline salinity, its technical process comprises: the cleaning of fresh ox and sheep bone is broken, steaming and decocting under high pressure, add protease hydrolyzed, acid adding, alkali adjust ph, filtering and concentrating, to dust drying, finished product packing, raw material sources are extensive, the polypeptide moiety of finished product can reach 90%, improve degree of hydrolysis, can be used for preparation plurality kinds of health care food, be applicable to the method for polypeptide in Enzymatic Extraction ox, sheep animal skeleton, but polypeptide moiety can not be improved as much as possible for valuable bone raw material, cause unnecessary waste.
To sum up, prior art polypeptide extracting method single, extraction efficiency is not high, cause waste, applicable situation is restricted, and the polypeptide of preparation especially small active peptides is difficult to use in peptide medicament and peptide class healthcare products, to meeting growing healthcare products and drug market is not significant.
Summary of the invention
The present invention is for effectively to solve the problem, the invention provides a kind of method that micromolecule active polypeptide especially extracts micromolecule polypeptide from yak, the method can comprehensive advantage, the extraction efficiency of great raising micromolecule active polypeptide, and the method can be applicable to the extraction of multiple bone polypeptide.
Technical scheme of the present invention is a kind of method extracting yak small-molecular peptides from yak bone, in turn includes the following steps: (1) prepares starter; (2) yak bone pre-treatment; (3) elementary fermentable; (4) the elementary yak albumen of electrolysis; (5) alcohol activates the preparation of pancreas slurry; (6) enzymolysis process; (7) last handling process.
Further step (1) is prepared in starter process, the liquid culture of useful bacterial strain is used to prepare starter, comprise subtilis, bifid Helicobacter pylori, lactobacillus rhamnosus, photosynthetic bacteria, Bacillus licheniformis, also add nutrient substance, described nutrient substance is wort;
In the pre-treatment of further step (2) yak bone, the method for steaming and decocting under high pressure is adopted to obtain bone soup; In step (3) elementary fermentable, add agent of promoting production, described agent of promoting production is phytic acid matter, and fermentation obtains the yak albumen of elementary fermentable, i.e. elementary yak albumen;
In the elementary yak albumen of further step (4) electrolysis, described elementary yak albumen is carried out electrolysis, obtain yak oligopeptides bone soup; Step (5) alcohol activates in the preparation of pancreas slurry: pancreas does proteolytic enzyme source, adopts edible ethanol one-step activation to form pancreatin, forms alcohol and activates pancreas slurry;
In further step (6) enzymolysis process: yak oligopeptides bone soup, alcohol activate pancreas slurry and carries out enzymolysis according to certain mass ratio, divide and add described alcohol activation pancreas slurry for three times, obtain elementary yak small-molecular peptides; In step (7) last handling process: obtain yak small-molecular peptides through aftertreatment.
A kind of micromolecule active polypeptide provided by the invention especially extracts the method for micromolecule polypeptide from yak, step is as follows more specifically: (1) prepares starter: use the liquid culture of useful bacterial strain to prepare starter, take subtilis 10 weight part, bifid Helicobacter pylori 10 weight part, lactobacillus rhamnosus 40 weight part, wort 15 weight part, photosynthetic bacteria 20 weight part, Bacillus licheniformis 5 weight part mixes, and be placed to and be loaded with in the triangular flask of substratum, carry out compound criteria, described substratum is nutrient agar, the temperature of carrying out cultivating in described triangular flask is 37 DEG C, 18-20h is cultivated under 200-250rpm/min, thalline or spore concentration is made to reach 5 × 10
13individual/liter (CFU/L), obtained starter, (2) yak bone pre-treatment: fresh yak bone is broken, cleaning, in digester, adopt temperature to be more than 130 DEG C temperature, pressure 0.17-0.18MPa steaming and decocting under high pressure 300-400min, bone slag is discharged, soup oil imports multi_layer extraction in soup oil tank, obtains bone soup, (3) elementary fermentable: the bone soup that step (2) obtains is proceeded to in primary fermentation production tank, and add the agent of promoting production of the 0.5-0.8% of described bone soup weight, described agent of promoting production is phytic acid matter, at the occurs at low temperatures ferment 8-10h of 35-38 DEG C, then be warming up to 160-200 DEG C and continue fermentation 5-8h, obtain the yak albumen of elementary fermentable, i.e. elementary yak albumen, (4) the elementary yak albumen of electrolysis, described elementary yak albumen is carried out electrolysis, use conventional protein electrolytic process, in electrolytic process, bath voltage is 20-50mV, and electrolysis time is 3-5h, get three sample testings, when the molecular weight ranges of the yak peptide in each sample is all no more than 800 dalton, then stop electrolysis, obtain yak oligopeptides bone soup, (5) alcohol activates the preparation of pancreas slurry: it is as follows that alcohol activates pancreas slurry preparation process: in pancreatin activator appliance, put into clean pancreas and do proteolytic enzyme source, edible ethanol one-step activation is adopted to form pancreatin, the add-on of described edible ethanol is 2 times of described pancreatin quality, the volumetric concentration of described edible ethanol is 45%, described activationary time is 60-80h, and activationary temperature is 6-8 DEG C, becomes alcohol and activates pancreas slurry, (6) enzymolysis process: described yak oligopeptides bone soup is imported in hydrolytic decomposition pot, then divide and add described alcohol activation pancreas slurry for three times, the mass ratio that the described alcohol added for three times activates pancreas slurry is 1: 1: 4, add when described enzymolysis starts for the first time, second time adds when described enzymolysis carries out 1h, it is that described enzymolysis adds when carrying out 2h that third time adds, described enzymolysis total time is 7h, what described alcohol activation pancreas was starched adds the 1/9-1/5 that quality is yak oligopeptides bone soup quality, keeps pH in hydrolytic decomposition pot to be 6.7-7.0 in described enzymolysis process, in described enzymolysis process first time add alcohol activate pancreas slurry after to second time add alcohol activate pancreas slurry before hydrolysis temperature be 60 DEG C, second time add alcohol activate pancreas slurry after hydrolysis temperature be 45 DEG C, obtain elementary yak small-molecular peptides, (7) last handling process: through described enzymolysis process, namely activates the hydrolysis of pancreas slurry in hydrolytic decomposition pot through described alcohol, becomes the mixing liquid that hydrolysis is final, after filtering, import concentration tank concentrate, thickening temperature is 50 DEG C, concentrated at least 2h, lower the temperature afterwards, when being cooled to 30 DEG C, when mensuration degree Beaume is not less than 30, stop concentrated, by concentrated solution boiling sterilization, drying of dusting, moisture content is down to less than 2.5%, send into storage tank, pack, obtain yak small-molecular peptides.
Preferably, step (1) is prepared in starter process, and the temperature of carrying out cultivating in described triangular flask is 37 DEG C, under 200rpm/min, cultivate 20h; Step (2) yak bone pre-treatment: in digester, adopts temperature to be 150 DEG C, pressure 0.17MPa steaming and decocting under high pressure 400min.
Preferably, in step (3) elementary fermentable, add the phytic acid matter of 0.8% of described bone soup weight, at the occurs at low temperatures ferment 8h of 38 DEG C, be then warming up to 165 DEG C and continue fermentation 7h, obtain elementary yak albumen; In the elementary yak albumen of step (4) electrolysis, bath voltage is 30mV, and electrolysis time is 4h.
Preferably, step (5) alcohol activates in the preparation of pancreas slurry: described activationary time is 60h, and activationary temperature is 6 DEG C, becomes alcohol and activates pancreas slurry; In step (6) enzymolysis process: described alcohol activates add that quality is yak oligopeptides bone soup quality 1/7 of pancreas slurry, keeps pH in hydrolytic decomposition pot to be 6.8 in described enzymolysis process; In step (7) last handling process: concentrated 3h, cooling, when being cooled to 30 DEG C, stops concentrated when measuring degree Beaume 35.
Preferably, yak finished product content of peptides is close to 100%, and the molecular weight of described yak small-molecular peptides is that the daltonian accounting of 0-480 is no less than 88%.
Preferably, the molecular weight of described yak small-molecular peptides is the daltonian accounting of 0-480 be the daltonian accounting of 89.62%, 480-1000 is 7.38%, and being greater than 1000 daltonian accountings is 3%; Color and luster is faint yellow.
Present invention also offers the application of above-mentioned yak small-molecular peptides, described yak small-molecular peptides is applicable to peptide medicament and peptide class healthcare products.
Preferably, described yak small-molecular peptides is applicable to diagnosis peptide, peptide vaccine, antibacterial active peptide, cytokine simulating peptide, antiviral peptide and/or natineoplaston.
The present invention has following useful effect:
The extracting method that the object of the invention is to carry yak micromolecule active polypeptide is a kind of integrated approach, and use range is extensive, not only may be used for extracting polypeptide in yak bone, can also be used for extracting micromolecule active polypeptide in other animal skeleton.
Yak micromolecule active polypeptide comparison of ingredients prepared by the present invention is pure, be far superior to the polypeptide prepared in prior art, yak micromolecule active polypeptide content prepared by the present invention is close to 100%, the molecular weight of described yak small-molecular peptides is that the daltonian accounting of 0-480 is no less than 87.5%, the molecular weight of concrete described yak small-molecular peptides is the daltonian accounting of 0-480 is 88.54%, the daltonian accounting of 480-1000 is 9.58%, being greater than 1000 daltonian accountings is 1.88%, and color and luster is faint yellow.
Yak small-molecular peptides prepared by the present invention, can be applied to peptide medicament and peptide class healthcare products, and range of application is relatively more extensive, such as, for diagnosis peptide, peptide vaccine, antibacterial active peptide, cytokine simulating peptide, antiviral peptide and/or natineoplaston etc.
Yak small-molecular peptides prepared by the present invention meets the use needs of growing medicine and healthcare products, can prepare use on a large scale safely.
Embodiment
Embodiment 1
The preparation method of yak small-molecular peptides, in turn include the following steps: (1) prepares starter: use the liquid culture of useful bacterial strain to prepare starter, take subtilis 10 weight part, bifid Helicobacter pylori 10 weight part, lactobacillus rhamnosus 40 weight part, wort 15 weight part, photosynthetic bacteria 20 weight part, Bacillus licheniformis 5 weight part mixes, and be placed to and be loaded with in the triangular flask of substratum, carry out compound criteria, described substratum is nutrient agar, the temperature of carrying out cultivating in described triangular flask is 37 DEG C, 20h is cultivated under 200rpm/min, thalline or spore concentration is made to reach 5 × 10
13individual/liter (CFU/L), obtained starter, (2) yak bone pre-treatment: fresh yak bone is broken, cleaning, in digester, employing temperature is the temperature of 150 DEG C, pressure 0.17MPa steaming and decocting under high pressure 400min, and bone slag is discharged, and soup oil imports multi_layer extraction in soup oil tank, obtains bone soup, (3) elementary fermentable: the bone soup that step (2) obtains is proceeded to in primary fermentation production tank, and add the agent of promoting production of 0.8% of described bone soup weight, described agent of promoting production is phytic acid matter, at the occurs at low temperatures ferment 10h of 38 DEG C, then be warming up to 200 DEG C and continue fermentation 8h, obtain the yak albumen of elementary fermentable, i.e. elementary yak albumen, (4) the elementary yak albumen of electrolysis, described elementary yak albumen is carried out electrolysis, use conventional protein electrolytic process, in electrolytic process, bath voltage is 50mV, and electrolysis time is 5h, get three sample testings, when the molecular weight ranges of the yak peptide in each sample is all no more than 800 dalton, then stop electrolysis, obtain yak oligopeptides bone soup, (5) alcohol activates the preparation of pancreas slurry: it is as follows that alcohol activates pancreas slurry preparation process: in pancreatin activator appliance, put into clean pancreas and do proteolytic enzyme source, edible ethanol one-step activation is adopted to form pancreatin, the add-on of described edible ethanol is 2 times of described pancreatin quality, the volumetric concentration of described edible ethanol is 45%, described activationary time is 80h, and activationary temperature is 8 DEG C, becomes alcohol and activates pancreas slurry, (6) enzymolysis process: described yak oligopeptides bone soup is imported in hydrolytic decomposition pot, then divide and add described alcohol activation pancreas slurry for three times, the mass ratio that the described alcohol added for three times activates pancreas slurry is 1: 1: 4, add when described enzymolysis starts for the first time, second time adds when described enzymolysis carries out 1h, it is that described enzymolysis adds when carrying out 2h that third time adds, described enzymolysis total time is 7h, described alcohol activates add that quality is yak oligopeptides bone soup quality 1/5 of pancreas slurry, keeps pH in hydrolytic decomposition pot to be 6.7 in described enzymolysis process, in described enzymolysis process first time add alcohol activate pancreas slurry after to second time add alcohol activate pancreas slurry before hydrolysis temperature be 60 DEG C, second time add alcohol activate pancreas slurry after hydrolysis temperature be 45 DEG C, obtain elementary yak small-molecular peptides, (7) last handling process: through described enzymolysis process, namely activates the hydrolysis of pancreas slurry in hydrolytic decomposition pot through described alcohol, becomes the mixing liquid that hydrolysis is final, after filtering, import concentration tank concentrate, thickening temperature is 50 DEG C, concentrated 3h, lower the temperature afterwards, when being cooled to 30 DEG C, when measuring degree Beaume 40, stop concentrated, by concentrated solution boiling sterilization, drying of dusting, moisture content is down to 1.5%, send into storage tank, pack, obtain yak small-molecular peptides.
The yak small-molecular peptides that above-described embodiment 1 prepares pure, be far superior to the polypeptide prepared in prior art, content is close to 100%, the molecular weight of yak small-molecular peptides is the daltonian accounting of 0-480 is 88.54%, the daltonian accounting of 480-1000 is 9.58%, being greater than 1000 daltonian accountings is 1.88%, and color and luster is faint yellow, can be applied to peptide pharmaceutical products and/or healthcare products.
Embodiment 2
The preparation method of yak small-molecular peptides, in turn include the following steps: (1) prepares starter: use the liquid culture of useful bacterial strain to prepare starter, take subtilis 10 weight part, bifid Helicobacter pylori 10 weight part, lactobacillus rhamnosus 40 weight part, wort 15 weight part, photosynthetic bacteria 20 weight part, Bacillus licheniformis 5 weight part mixes, and be placed to and be loaded with in the triangular flask of substratum, carry out compound criteria, described substratum is nutrient agar, the temperature of carrying out cultivating in described triangular flask is 37 DEG C, 19h is cultivated under 220rpm/min, thalline or spore concentration is made to reach 5 × 10
13individual/liter (CFU/L), obtained starter, (2) yak bone pre-treatment: fresh yak bone is broken, cleaning, in digester, employing temperature is the temperature of 140 DEG C, pressure 0.17MPa steaming and decocting under high pressure 350min, and bone slag is discharged, and soup oil imports multi_layer extraction in soup oil tank, obtains bone soup, (3) elementary fermentable: the bone soup that step (2) obtains is proceeded to in primary fermentation production tank, and add the agent of promoting production of 0.6% of described bone soup weight, described agent of promoting production is phytic acid matter, at the occurs at low temperatures ferment 9h of 37 DEG C, then be warming up to 190 DEG C and continue fermentation 7h, obtain the yak albumen of elementary fermentable, i.e. elementary yak albumen.(4) the elementary yak albumen of electrolysis, described elementary yak albumen is carried out electrolysis, use conventional protein electrolytic process, in electrolytic process, bath voltage is 40mV, and electrolysis time is 4h, get three sample testings, when the molecular weight ranges of the yak peptide in each sample is all no more than 800 dalton, then stop electrolysis, obtain yak oligopeptides bone soup.(5) alcohol activates the preparation of pancreas slurry: it is as follows that alcohol activates pancreas slurry preparation process: in pancreatin activator appliance, put into clean pancreas and do proteolytic enzyme source, edible ethanol one-step activation is adopted to form pancreatin, the add-on of described edible ethanol is 2 times of described pancreatin quality, the volumetric concentration of described edible ethanol is 45%, described activationary time is 70h, and activationary temperature is 6 DEG C, becomes alcohol and activates pancreas slurry.(6) enzymolysis process: described yak oligopeptides bone soup is imported in hydrolytic decomposition pot, then divide and add described alcohol activation pancreas slurry for three times, the mass ratio that the described alcohol added for three times activates pancreas slurry is 1: 1: 4, add when described enzymolysis starts for the first time, second time adds when described enzymolysis carries out 1h, it is that described enzymolysis adds when carrying out 2h that third time adds, described enzymolysis total time is 7h, described alcohol activates add that quality is yak oligopeptides bone soup quality 1/5 of pancreas slurry, keeps pH in hydrolytic decomposition pot to be 7.0 in described enzymolysis process; In described enzymolysis process first time add alcohol activate pancreas slurry after to second time add alcohol activate pancreas slurry before hydrolysis temperature be 60 DEG C, second time add alcohol activate pancreas slurry after hydrolysis temperature be 45 DEG C, obtain elementary yak small-molecular peptides.(7) last handling process: through described enzymolysis process, namely activates the hydrolysis of pancreas slurry in hydrolytic decomposition pot through described alcohol, becomes the mixing liquid that hydrolysis is final, after filtering, import concentration tank concentrate, thickening temperature is 50 DEG C, concentrated at least 2h, lower the temperature afterwards, when being cooled to 30 DEG C, when measuring degree Beaume 35, stop concentrated, by concentrated solution boiling sterilization, drying of dusting, moisture content is down to 2.3%, send into storage tank, pack, obtain yak small-molecular peptides.
The yak small-molecular peptides that above-described embodiment 2 prepares pure, be far superior to the polypeptide prepared in prior art, content is close to 100%, the molecular weight of yak small-molecular peptides is the daltonian accounting of 0-480 is 87.9%, the daltonian accounting of 480-1000 is 10.24%, being greater than 1000 daltonian accountings is 1.86%, and color and luster is faint yellow, can be applied to peptide pharmaceutical products and/or healthcare products.
Above-described embodiment, only for technical conceive of the present invention and feature are described, its object is to person skilled in the art can be understood content of the present invention and implement according to this, can not limit the scope of the invention with this.All equivalences done according to spirit of the present invention change or modify, and all should be encompassed within protection scope of the present invention.
Claims (10)
1. from yak bone, extract a method for yak small-molecular peptides, it is characterized in that, in turn include the following steps: (1) prepares starter; (2) yak bone pre-treatment; (3) elementary fermentable; (4) the elementary yak albumen of electrolysis; (5) alcohol activates the preparation of pancreas slurry; (6) enzymolysis process; (7) last handling process.
2. the method for claim 1, is characterized in that,
Step (1) is prepared in starter process, the liquid culture of useful bacterial strain is used to prepare starter, comprise subtilis, bifid Helicobacter pylori, lactobacillus rhamnosus, photosynthetic bacteria, Bacillus licheniformis, also add nutrient substance, described nutrient substance is wort;
In the pre-treatment of step (2) yak bone, the method for steaming and decocting under high pressure is adopted to obtain bone soup;
In step (3) elementary fermentable, add agent of promoting production, described agent of promoting production is phytic acid matter, and fermentation obtains the yak albumen of elementary fermentable, i.e. elementary yak albumen;
In the elementary yak albumen of step (4) electrolysis, described elementary yak albumen is carried out electrolysis, obtain yak oligopeptides bone soup;
Step (5) alcohol activates in the preparation of pancreas slurry: pancreas does proteolytic enzyme source, adopts edible ethanol one-step activation to form pancreatin, forms alcohol and activates pancreas slurry;
In step (6) enzymolysis process: yak oligopeptides bone soup, alcohol activate pancreas slurry and carries out enzymolysis according to certain mass ratio, divide and add described alcohol activation pancreas slurry for three times, obtain elementary yak small-molecular peptides;
In step (7) last handling process: obtain yak small-molecular peptides through aftertreatment.
3. the method for claim 1, is characterized in that, concrete grammar is as follows:
(1) starter is prepared: use the liquid culture of useful bacterial strain to prepare starter, take subtilis 10 weight part, bifid Helicobacter pylori 10 weight part, lactobacillus rhamnosus 40 weight part, wort 15 weight part, photosynthetic bacteria 20 weight part, Bacillus licheniformis 5 weight part mixes, and be placed to and be loaded with in the triangular flask of substratum, carry out compound criteria, described substratum is nutrient agar, the temperature of carrying out cultivating in described triangular flask is 37 DEG C, 18-20h is cultivated under 200-250rpm/min, thalline or spore concentration is made to reach 5 × 10
13individual/liter (CFU/L), obtained starter.
(2) yak bone pre-treatment: fresh yak bone is broken, cleaning, in digester, adopt temperature to be more than 130 DEG C temperature, pressure 0.17-0.18MPa steaming and decocting under high pressure 300-400min, bone slag is discharged, soup oil imports multi_layer extraction in soup oil tank, obtains bone soup
(3) elementary fermentable: the bone soup that step (2) obtains is proceeded to in primary fermentation production tank, and add the agent of promoting production of the 0.5-0.8% of described bone soup weight, described agent of promoting production is phytic acid matter, at the occurs at low temperatures ferment 8-10h of 35-38 DEG C, then be warming up to 160-200 DEG C and continue fermentation 5-8h, obtain the yak albumen of elementary fermentable, i.e. elementary yak albumen.
(4) the elementary yak albumen of electrolysis, described elementary yak albumen is carried out electrolysis, use conventional protein electrolytic process, in electrolytic process, bath voltage is 20-50mV, and electrolysis time is 3-5h, get three sample testings, when the molecular weight ranges of the yak peptide in each sample is all no more than 800 dalton, then stop electrolysis, obtain yak oligopeptides bone soup.
(5) alcohol activates the preparation of pancreas slurry: it is as follows that alcohol activates pancreas slurry preparation process: in pancreatin activator appliance, put into clean pancreas and do proteolytic enzyme source, edible ethanol one-step activation is adopted to form pancreatin, the add-on of described edible ethanol is 2 times of described pancreatin quality, the volumetric concentration of described edible ethanol is 45%, described activationary time is 60-80h, and activationary temperature is 6-8 DEG C, becomes alcohol and activates pancreas slurry.
(6) enzymolysis process: described yak oligopeptides bone soup is imported in hydrolytic decomposition pot, then divide and add described alcohol activation pancreas slurry for three times, the mass ratio that the described alcohol added for three times activates pancreas slurry is 1: 1: 4, add when described enzymolysis starts for the first time, second time adds when described enzymolysis carries out 1h, it is that described enzymolysis adds when carrying out 2h that third time adds, described enzymolysis total time is 7h, what described alcohol activation pancreas was starched adds the 1/9-1/5 that quality is yak oligopeptides bone soup quality, keeps pH in hydrolytic decomposition pot to be 6.7-7.0 in described enzymolysis process; In described enzymolysis process first time add alcohol activate pancreas slurry after to second time add alcohol activate pancreas slurry before hydrolysis temperature be 60 DEG C, second time add alcohol activate pancreas slurry after hydrolysis temperature be 45 DEG C, obtain elementary yak small-molecular peptides.
(7) last handling process: through described enzymolysis process, namely activates the hydrolysis of pancreas slurry in hydrolytic decomposition pot through described alcohol, becomes the mixing liquid that hydrolysis is final, after filtering, import concentration tank concentrate, thickening temperature is 50 DEG C, concentrated at least 2h, lower the temperature afterwards, when being cooled to 30 DEG C, when mensuration degree Beaume is not less than 30, stop concentrated, by concentrated solution boiling sterilization, drying of dusting, moisture content is down to less than 2.5%, send into storage tank, pack, obtain yak small-molecular peptides.
4. method as claimed in claim 3, it is characterized in that, step (1) is prepared in starter process, and the temperature of carrying out cultivating in described triangular flask is 37 DEG C, under 200rpm/min, cultivate 20h; Step (2) yak bone pre-treatment: in digester, adopts temperature to be 150 DEG C, pressure 0.17MPa steaming and decocting under high pressure 400min.
5. method as claimed in claim 3, is characterized in that, in step (3) elementary fermentable, add the phytic acid matter of 0.8% of described bone soup weight, at the occurs at low temperatures ferment 8h of 38 DEG C, be then warming up to 165 DEG C and continue fermentation 7h, obtain elementary yak albumen; In the elementary yak albumen of step (4) electrolysis, bath voltage is 30mV, and electrolysis time is 4h.
6. method as claimed in claim 3, is characterized in that, step (5) alcohol activates in the preparation of pancreas slurry: described activationary time is 60h, and activationary temperature is 6 DEG C, becomes alcohol and activates pancreas slurry; In step (6) enzymolysis process: described alcohol activates add that quality is yak oligopeptides bone soup quality 1/7 of pancreas slurry, keeps pH in hydrolytic decomposition pot to be 6.8 in described enzymolysis process; In step (7) last handling process: concentrated 3h, cooling, when being cooled to 30 DEG C, stops concentrated when measuring degree Beaume 35.
7. the yak small-molecular peptides that as described in any one of claim 1-6 prepared by method, is characterized in that, yak finished product content of peptides is close to 100%, and the molecular weight of described yak small-molecular peptides is that the daltonian accounting of 0-480 is no less than 87.5%.
8. the yak small-molecular peptides prepared of method as claimed in claim 7, it is characterized in that, the molecular weight of described yak small-molecular peptides is the daltonian accounting of 0-480 be the daltonian accounting of 88.54%, 480-1000 is 9.58%, and being greater than 1000 daltonian accountings is 1.88%; Color and luster is faint yellow.
9. the application of yak small-molecular peptides as described in any one of claim 7-8, it is characterized in that, described yak small-molecular peptides is applicable to peptide medicament and peptide class healthcare products.
10. apply as claimed in claim 9, it is characterized in that, described yak small-molecular peptides is applicable to diagnosis peptide, peptide vaccine, antibacterial active peptide, cytokine simulating peptide, antiviral peptide and/or natineoplaston.
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| CN106309372A (en) * | 2016-08-31 | 2017-01-11 | 湖北瑞邦生物科技有限公司 | Bovine bone polypeptide sustained-release preparation and preparation method thereof |
| CN109125704A (en) * | 2018-11-01 | 2019-01-04 | 北京市蜂业公司 | Royal jelly Yak Bone small-molecular peptides and its preparation method and application |
| CN110403054A (en) * | 2019-06-06 | 2019-11-05 | 光泽县泽农生物科技有限公司 | A method of improving chicken polypeptide absorption rate |
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| CN106309372A (en) * | 2016-08-31 | 2017-01-11 | 湖北瑞邦生物科技有限公司 | Bovine bone polypeptide sustained-release preparation and preparation method thereof |
| CN106309372B (en) * | 2016-08-31 | 2019-12-24 | 湖北瑞邦生物科技有限公司 | Bovine bone polypeptide sustained release agent and preparation method thereof |
| CN109125704A (en) * | 2018-11-01 | 2019-01-04 | 北京市蜂业公司 | Royal jelly Yak Bone small-molecular peptides and its preparation method and application |
| CN110403054A (en) * | 2019-06-06 | 2019-11-05 | 光泽县泽农生物科技有限公司 | A method of improving chicken polypeptide absorption rate |
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