CN108396051A - A kind of industrialized producing technology of sheep placenta lamb albumen oligopeptide - Google Patents
A kind of industrialized producing technology of sheep placenta lamb albumen oligopeptide Download PDFInfo
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- CN108396051A CN108396051A CN201810262025.7A CN201810262025A CN108396051A CN 108396051 A CN108396051 A CN 108396051A CN 201810262025 A CN201810262025 A CN 201810262025A CN 108396051 A CN108396051 A CN 108396051A
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- 241001494479 Pecora Species 0.000 title claims abstract description 56
- 210000002826 placenta Anatomy 0.000 title claims abstract description 56
- 108010038807 Oligopeptides Proteins 0.000 title claims abstract description 51
- 102000015636 Oligopeptides Human genes 0.000 title claims abstract description 51
- 238000005516 engineering process Methods 0.000 title claims abstract description 10
- 238000000108 ultra-filtration Methods 0.000 claims abstract description 31
- 238000001728 nano-filtration Methods 0.000 claims abstract description 30
- 238000001914 filtration Methods 0.000 claims abstract description 27
- 238000004108 freeze drying Methods 0.000 claims abstract description 20
- 238000011049 filling Methods 0.000 claims abstract description 18
- 238000000034 method Methods 0.000 claims abstract description 18
- 239000002994 raw material Substances 0.000 claims abstract description 17
- 238000005238 degreasing Methods 0.000 claims abstract description 14
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- 239000002002 slurry Substances 0.000 claims description 19
- 239000012141 concentrate Substances 0.000 claims description 17
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 17
- 239000012530 fluid Substances 0.000 claims description 16
- 239000007788 liquid Substances 0.000 claims description 16
- 238000010438 heat treatment Methods 0.000 claims description 13
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 12
- 238000010792 warming Methods 0.000 claims description 12
- JGSARLDLIJGVTE-UHFFFAOYSA-N 3,3-dimethyl-7-oxo-6-[(2-phenylacetyl)amino]-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid Chemical compound O=C1N2C(C(O)=O)C(C)(C)SC2C1NC(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-UHFFFAOYSA-N 0.000 claims description 9
- 239000000843 powder Substances 0.000 claims description 9
- 239000000047 product Substances 0.000 claims description 9
- 238000001223 reverse osmosis Methods 0.000 claims description 9
- 102000008186 Collagen Human genes 0.000 claims description 8
- 108010035532 Collagen Proteins 0.000 claims description 8
- 229920001436 collagen Polymers 0.000 claims description 8
- 239000012634 fragment Substances 0.000 claims description 8
- 239000000706 filtrate Substances 0.000 claims description 7
- 238000002372 labelling Methods 0.000 claims description 7
- 150000003839 salts Chemical class 0.000 claims description 7
- 239000012528 membrane Substances 0.000 claims description 6
- 102000004169 proteins and genes Human genes 0.000 claims description 6
- 108090000623 proteins and genes Proteins 0.000 claims description 6
- 239000012465 retentate Substances 0.000 claims description 6
- 239000011780 sodium chloride Substances 0.000 claims description 6
- 230000009849 deactivation Effects 0.000 claims description 5
- 238000000227 grinding Methods 0.000 claims description 5
- 239000012510 hollow fiber Substances 0.000 claims description 5
- 238000010828 elution Methods 0.000 claims description 4
- 239000002245 particle Substances 0.000 claims description 4
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- 239000004744 fabric Substances 0.000 claims description 3
- 238000005192 partition Methods 0.000 claims description 3
- 239000004033 plastic Substances 0.000 claims description 3
- 238000007711 solidification Methods 0.000 claims description 3
- 230000008023 solidification Effects 0.000 claims description 3
- 239000012065 filter cake Substances 0.000 claims description 2
- 230000017854 proteolysis Effects 0.000 claims description 2
- 230000036541 health Effects 0.000 abstract description 5
- 239000003795 chemical substances by application Substances 0.000 abstract description 2
- 239000000490 cosmetic additive Substances 0.000 abstract description 2
- 238000013461 design Methods 0.000 abstract description 2
- 239000002778 food additive Substances 0.000 abstract 1
- 235000013373 food additive Nutrition 0.000 abstract 1
- 230000007062 hydrolysis Effects 0.000 abstract 1
- 238000006460 hydrolysis reaction Methods 0.000 abstract 1
- 238000011031 large-scale manufacturing process Methods 0.000 abstract 1
- 239000002417 nutraceutical Substances 0.000 abstract 1
- 235000021436 nutraceutical agent Nutrition 0.000 abstract 1
- 238000004519 manufacturing process Methods 0.000 description 8
- 230000006872 improvement Effects 0.000 description 6
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- 102000004196 processed proteins & peptides Human genes 0.000 description 4
- MVORZMQFXBLMHM-QWRGUYRKSA-N Gly-His-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CN=CN1 MVORZMQFXBLMHM-QWRGUYRKSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
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- 108010038983 glycyl-histidyl-lysine Proteins 0.000 description 3
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- 238000010257 thawing Methods 0.000 description 3
- 206010054949 Metaplasia Diseases 0.000 description 2
- 230000033228 biological regulation Effects 0.000 description 2
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- 238000007789 sealing Methods 0.000 description 2
- 230000006641 stabilisation Effects 0.000 description 2
- 238000011105 stabilization Methods 0.000 description 2
- 238000012795 verification Methods 0.000 description 2
- 210000001835 viscera Anatomy 0.000 description 2
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- 102000014150 Interferons Human genes 0.000 description 1
- 108010050904 Interferons Proteins 0.000 description 1
- 241000283903 Ovis aries Species 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 102000007079 Peptide Fragments Human genes 0.000 description 1
- 108010033276 Peptide Fragments Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
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- 235000013305 food Nutrition 0.000 description 1
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- 229940088597 hormone Drugs 0.000 description 1
- 238000007654 immersion Methods 0.000 description 1
- 229940079322 interferon Drugs 0.000 description 1
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- 244000144972 livestock Species 0.000 description 1
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- 238000012216 screening Methods 0.000 description 1
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- 238000005092 sublimation method Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
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- 239000012588 trypsin Substances 0.000 description 1
- 238000009777 vacuum freeze-drying Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
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- 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
-
- 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 or cold insoluble globulin [CIG]
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Abstract
The present invention provides a kind of industrialized producing technologies of sheep placenta lamb albumen oligopeptide.It is as follows：1, raw material, 2, slice, 3, impregnate thaw, 4, rub, 5, degreasing, 6, defibrination, 7, saltout, 8, press filtration, 9, enzymolysis, 10, ultrafiltration oligopeptide, 11, nanofiltration retention concentration, desalination, removing impurities matter, 12, degerming, 13, it is filling, 14, freeze-drying, 15, pressure bottle cap and plug, label.The method of the present invention is simple for process, reasonable design, makes full use of the natural resources, it can be achieved that industrializing and large-scale production.The present invention uses food-grade albumen enzyme hydrolysis, safe, without side-effects；The present invention can be used as health products, function food additive or nutraceutical and directly take, while be also used as cosmetic additive agent safe handling.
Description
Technical field
The present invention relates to a kind of industrialized producing technologies of sheep placenta lamb albumen oligopeptide, belong to functional health care product or health care food
Product and cosmetic additive agent field.
Background technology
China is sheep raising big country in the world, and livestock on hand quantity, meat yield, production skin amount come world forefront.Sheep has more
The more lambs of tire breed fast feature, and sheep placenta lamb is resourceful, and potentiality to be exploited is big.The small active peptides extracted from sheep placenta lamb, contain
There are many antibody and interferon, also containing there are many hormone, polysaccharide, hematopoietin and phosphatide etc..With albumen oligopeptide
The market demand is growing, and develops the sheep placenta lamb albumen oligopeptide with the active highly effective and safe environmental protection of biological health and produces work
Skill all has significance to scientific research and practical application.Prepare the industrialization method research of the oligomeric Gly-His-Lys of sheep placenta lamb albumen
Invention, it can be achieved that utilization to sheep placenta lamb Deep-Resource time.
Invention content
The present invention is in the above context and provides a kind of industry of output capacity height, the high sheep placenta lamb albumen oligopeptide of purity
Metaplasia production. art.
The present invention includes the following steps：1, raw material, 2, slice, 3, immersion defrosting, 4, rubbing, 5, degreasing, 6, defibrination, 7, salt
Analysis, 8, press filtration, 9, enzymolysis, 10, ultrafiltration oligopeptide, 11, nanofiltration retention concentration, desalination, removing impurities matter, 12, degerming, 13, it is filling,
14, freeze-drying, 15, pressure bottle cap and plug, labelling.It is digested using zymotechnic, collects the peptide fragment of suitable molecular weight, used
Ultrafiltration membrane technique prepares the sheep placenta lamb albumen oligopeptide for having a variety of physiological active functions by concentration, vacuum freezedrying.
A kind of industrialized producing technology of sheep placenta lamb albumen oligopeptide, its step are as follows：
1, raw material
Raw material, which is selected, completes bloodletting, remove internal organ, the ewe washed away breeds freeze it is completeSheep placenta lamb。
2, it is sliced
The sheep placenta lamb block burnishing surface frozen is lowered into the material path of slicer, it is made to be cut into the thin slice of 0.3-0.6cm thickness.
3, it impregnates and thaws
The sheep placenta lamb frozen is put into stainless steel rinse bath, pure water is injected, impregnates 48h-72 h, until sheep placenta lamb is at soft
State.
4, it rubs
It after pulling the sheep placenta lamb of the soft extraction of slice defrosting out water drenching, puts in meat grinder, it is 0.1- to be rubbed into granularity
The fragment of 0.2cm.
5, degreasing
By the degreaser soak degreasing 1-2d of the sheep placenta lamb of rubbing 1.0-2.0% acetums（It）, finally use 30-50 DEG C of temperature
Water rinses 2 times, obtains the sheep placenta lamb that degreasing is impregnated.
6, defibrination
Sheep placenta lamb after degreasing is worn into slurries with bone mud barreling, particle is become and refines slurries in 20~50 μm of sizes.
7, it saltouts
Fine grinding slurries obtained are adjusted into pH value 4.0~4.5, NaCl powder is added and saltouts, it is stirring while adding, make collagen egg
After white precipitation completely, standing 12~for 24 hours, obtain collagen mixed liquor.
8, press filtration
Mixed liquor is uniformly routed on sheet frame filter cloth, conveying mixed liquor starts 1~6h of press filtration, passes through the aperture regulation of outlet valve
Press filtration progress is controlled, filter cake is discarded, obtains transparent filtrate.
9, it digests
Under the conditions of being 9.0~9.5,50 DEG C~60 DEG C by the slurries adjusting pH value after press filtration, 1-3% trypsase is then added,
So that sheep placenta lamb proteolysis is reacted 8h~10h, is obtained after being stirred continuously with 30 turns/min in enzymolysis process mixed containing oligopeptide
Close liquid.
10, ultrafiltration oligopeptide
Enzymolysis liquid is subjected to classification ultrafiltration through reverse osmosis ultrafiltration unit, the ultrafilter of molecular weight 75KDa is first passed through, using molecule
Measure the ultrafilter filtering of 3KDa.The peptide of suitable molecular weight segment enters collector through film, and the peptide of large fragment continues and enzyme effect
It digests again, then ultrafiltration obtains oligopeptide filtered fluid.Its temperature is warming up to 80~95 DEG C of progress high temperature enzyme deactivations after ultrafiltration.
11, nanofiltration retention concentration, desalination, removing impurities matter
It is concentrated in the nanofiltration device that the molecular weight that albumen oligopeptide filtrate is pumped into reverse osmosis nanofiltration unit is 500Da.Again will
Concentrate adds same volume water elution 3-5 times, obtains and is concentrated into the minimum volume nanofiltration retention that water content only has 3-5% or so
Liquid.It will collect containing other substances and the nanofiltrations filtered fluid such as inorganic salts in raw material, carry out purified treatment, the nanofiltration retention of collection
Liquid is oligopeptide concentrate.
12, degerming
After concentrate is filtered using molecular weight 60-80KDa hollow fiber membrane ultrafiltration devices, enters back into 0.22 μm of micropore filter element and carried out
Filter, carries out degerming by way of retention.
13, filling
Oligopeptide concentrate after degerming is carried out filling with automatic filling machine, is fitted into and is cleaned in sterile 10ml cillin bottles, dosage
Control ensures the stabilization of net content within 5ml, per bottled amount 200mg, and pressurizes rubber stopper plug to state of partly jumping a queue, obtains
To canned product.
14, it is freeze-dried
（1）Pre-freeze cures
By canned concentrate cillin bottle, it is placed on the partition board in vacuum freeze drier dry storehouse, pre-freeze solidification temperature -30
~-45 DEG C, and 5~8h is kept, so that moisture in concentrate is fully freezed, it is ensured that no liquid exists.
（2）Lyophilization：Freeze dryer starts vacuum system, and pressure in storehouse is evacuated to 30~50pa, takes slow heating, with
1 DEG C of heating rate per hour is slowly warming up to -20~-25 DEG C, keeps the temperature 6~8h, then with 2 DEG C per hour of heating rate, delay
It is slow to be warming up to -10 DEG C~-12 DEG C, 3~6h is kept the temperature, lyophilization is carried out.
（3）Parsing-desiccation：After lyophilization, then with 3 DEG C per hour of heating rate, 40~50 DEG C are slowly warming up to, pressure
Power is 10~20Pa.6~8h is kept the temperature, parsing-desiccation freeze-drying is completed, is taken out after the plastic lid that pressurizes.
15, bottle cap and plug, labelling are pressed
Aseptically machinery pressure bottle cap and plug after freeze-drying, makes it post label into labelling machine.Purity of protein height is made
Oligopeptide component content up to 99.5%, relative molecular mass less than 1000Da is averagely more than 88% sheep placenta lamb oligopeptide finished product
Powder.
Compared with prior art, main advantages of the present invention are：
1, as a further improvement on the present invention, it is basic material that the present invention, which makes full use of sheep placenta lamb, is easy to get using raw material, cost
It is low, added value is high, the stringent selection of raw material substantially increases albumen oligopeptide output capacity, solves existing sheep placenta lamb resource and exists
Utilization rate and the low problem of added value.Sheep placenta lamb is sliced by present invention process process through strength, without straight under thawed state
Sheep placenta lamb will be frozen by, which connecing, thinly slices, and better assures that the quality of raw material.Keep raw material finer and smoother uniformly after the mill grinding of bone mud again,
Crushing effect is good, and advantage is created for the follow-up oligomeric peptide content for improving extraction.
2, as a further improvement on the present invention, the present invention is saltoutd in the extraction of albumen oligopeptide using NaCl powder
Method first collagen is cemented out, and after saltouing carry out plate and frame filter press press filtration, then to press filtration obtain slurries carry out
Enzymolysis.Single enzyme and compound protease screening verification have been carried out to the enzyme used in sheep placenta lamb collagen before enzymolysis, passed through verification
Selected the trypsase with complex enzyme function, and after taking appropriate enzymolysis, ultrafiltration, the enzymolysis and extraction side repeatedly digested again
Method destroys partial peptide key and forms the albumen oligopeptide that molecular weight is less than 1000Da, to not reaching the extracting solution of small-molecular peptides
Further enzymolysis is realized, high temperature enzyme deactivation is finally taken, enzymolysis process design ambassador's sheep placenta lamb collagen resolves into small
In the component content average out to 88% of the albumen oligopeptide of 1000Da molecular weight.Thus improve sheep placenta lamb protein active and utilization
Rate.
3, as a further improvement on the present invention, it is more than filtering method present invention employs reverse osmosis ultrafiltration unit, passes through
Macromolecular substances are retained repeatedly, and filtering, separation are achieved the purpose that through small-molecule substance.First pass through the super of molecular weight 75KDa
Filter filters, and is filtered using the ultrafilter of molecular weight 3KDa, and molecular cut off, will in 2000-100000Da protein peptides
1000Da small-molecular-weight segment oligopeptides below enter collector through film, and the peptide of large fragment continues and enzyme effect, then row is super
Filter.It ensure that the concentration output of oligomeric peptide product and making full use of for raw material.
4, as a further improvement on the present invention, the present invention uses molecular weight for the nanofiltration device of the reverse osmosis collecting and filtering apparatus of 500Da,
Oligopeptide is obtained by nanofiltration and retains concentrate, while making salt, impurity as nanofiltration filtered fluid is deviate from.It substantially increases
The purity of albumen oligopeptide so that albumen oligopeptide purity is up to 99.5%.Simultaneously by the nanofiltration containing salt, impurity of collection
Filtrate is pumped into biochemical treatment tank, carries out purified treatment, is discharged after up to standard so that production technology has reached environmentally protective standard.
5, as a further improvement on the present invention, the method that the present invention uses physics degerming, according to point of albumen oligopeptide
Son amount and bacteria molecule amount have selected molecular weight 60-80KDa hollow fiber membrane ultrafiltration devices to refilter, after filtering, into 0.22 micron
The small filter of degerming carries out degerming.The method of the degerming, which has, does not destroy active ingredient, does not increase other pollutions, efficient, degerming
Effect is good, effectively increases the shelf-life of the oligomeric peptide product of albumen.
6, as a further improvement on the present invention, the production of albumen oligopeptide of the present invention is in high-efficiency and continuous automated production
It is produced under system, entire technological process is mostly to be produced under sealing condition, such as filling using automatic filling machine progress, especially this hair
Bright freeze-drying takes slow heating technology, so that temperature in entire sublimation process is no more than eutectic point, ice crystal will not be caused to melt
Change and influence the quality of oligomeric Gly-His-Lys, ensure that the high activity of the oligomeric peptide product of albumen.Environment-friendly advantage of the present invention protrudes, and does not use
Toxic reagent, non-pollutant discharge reach clean manufacturing, environmentally protective, product safety health, with preferable industrialized production
Advanced and practicability.It is that sheep placenta lamb is reasonable using new approach is opened, is the albumen oligopeptide industrialization of sheep placenta lamb and scale
Metaplasia production provides effective process.
Specific implementation mode
Embodiment 1
A kind of industrialized producing technology of sheep placenta lamb albumen oligopeptide, its step are as follows：
1, raw material
Raw material, which is selected, completes bloodletting, removes internal organ, modifies, washes away, what the ewe after Dead weight bred freeze completeSheep placenta Lamb。
2, it is sliced
The sheep placenta lamb block burnishing surface frozen is lowered into the material path of slicer, makes its temperature at -12 DEG C to -6 DEG C, is pressed certainly
Dynamic charging button, pusher start pusher forward, start to cut, and adjust cylinder trip 70cm, air pressure 0.4-0.5Mpa, knife rotation
Speed is 435 turns/min, it is made to be cut into the thin slice of 0.3-0.6cm thickness.
3, it impregnates and thaws
The sheep placenta lamb frozen is put into stainless steel rinse bath, pure water is injected, height of water level will be more than sheep placenta lamb 10-15cm,
It stirs evenly, impregnates 48h-72 h, until sheep placenta lamb is at doughy state, while impregnating removing blood constituent.
4, it rubs
After pulling the sheep placenta lamb of the soft extraction of slice defrosting out water drenching, puts in meat grinder, rubbed, rubbed into granularity
For the fragment of 0.1-0.2cm.
5, degreasing
By the sheep placenta lamb of rubbing 1.0-2.0% acetum degreaser soak degreasing 1-2d, temperature is at 25-30 DEG C, per 2h-3h
It is sufficiently stirred 1 time, filters off supernatant liquid, it is primary to repeat degreasing as stated above, finally uses 30-50 DEG C of warm water rinse 2 times, obtains
Obtain the sheep placenta lamb that degreasing is impregnated.
6, defibrination
Sheep placenta lamb after degreasing is iterated through bone mud to grind 2-3 times, becomes particle in 20~50 μm of size slurries.Water is added to arrive
500-1000L water level lines impregnate 1~2d, remove soaking water, and soaked sheep placenta lamb is ground defibrination 1 time again in bone mud, is obtained
Finish milling slurry.
7, it saltouts
Fine grinding slurries obtained input 3000L is extracted into big tank, 100kg slurries add water 3000L, with 0.5mol/LHCl solution tune
PH value 4.0~4.5 is saved, is stirred evenly, NaCl powder is then added and saltouts, it is stirring while adding, keep 30r/min stirring slurries
Liquid, or so hour add 100kg sodium chloride.After collagen is precipitated completely, standing 12~for 24 hours, obtain collagen
Mixed serum.
8, press filtration
The sheet frame of plate and frame filter press is laid into No. 3927 filter clothes, 0.5% super-cell is added to the tank for filling enzymolysis liquid
In, it then stirs, is configured to the liquid material to be filtered of the enzymolysis liquid mixed liquor added with filter aid, it is later that mixed liquor is uniform
Be routed on sheet frame filter cloth, slowly open filter pressing pump inlet valve, outlet valve, start press filtration relay pump, pressure be 0.1~
1.0MPa, conveying mixed liquor start press filtration, control press filtration progress by the aperture regulation of outlet valve, filter is discarded after the completion of press filtration
Cake obtains transparent filtering slurries.
9, it digests
Slurries after press filtration are added in 2500L enzymatic vessels, add 7L water, 600L slurries about to add water 1400L according to every 3L slurries,
It is 7.0~9.0 to add sodium hydroxide and adjust the pH value of slurries, and is heated under the conditions of 50 DEG C~60 DEG C, then by 1-3%
Trypsase is added in the enzymatic vessel, makes sheep placenta lamb albumen and tryptose enzyme digestion reaction 8h~10h, in enzymolysis process with
30 turns/min is stirred continuously, and the mixed liquor containing a large amount of oligopeptides is obtained by enzymolysis.
10, ultrafiltration oligopeptide
Oligopeptide mixed liquor after enzymolysis is subjected to classification ultrafiltration through reverse osmosis ultrafiltration unit, first passes through the super of molecular weight 75KDa
Filter filters, and is filtered using the ultrafilter of molecular weight 3KDa.Ensure pressure 3.0-5.0MPa, flow velocity 1000- in ultra-filtration process
1200L/h adds water 800L ultrafiltration 2-3 times every time, and the peptide of suitable molecular weight segment enters collector through film after ultrafiltration, retention
The peptide of large fragment continues and enzyme effect digests again, then ultrafiltration obtains the peptide of suitable molecular weight segment.It is low that albumen is obtained repeatedly
Poly- peptide filtered fluid.Filtered fluid temperature is warming up to 80~95 DEG C of progress high temperature enzyme deactivations, 10~25min after ultrafiltration.
11, nanofiltration retention concentration, desalination, removing impurities matter
In the nanofiltration device that the molecular weight that albumen oligopeptide filtrate is pumped into reverse osmosis nanofiltration unit is 500Da, holding pressure 2.5~
3.5MPa carries out reverse osmosis nanofiltration concentration under conditions of temperature is 15~45 DEG C, until nanofiltration retentate fluid is concentrated into water content only
There is the minimum volume of 3-5% or so, then concentrate is added into same volume water elution 3-5 times, while obtaining and being concentrated into water content only
There is the minimum volume nanofiltration retentate fluid of 3-5% or so.It is collected nanofiltration filtered fluid and nanofiltration retentate fluid respectively, collection is contained
The nanofiltrations filtered fluids such as other impurities substance and inorganic salts in raw material are pumped into biochemical treatment tank and carry out purified treatment, heel row up to standard
It puts, the nanofiltration retentate fluid of collection is sheep placenta lamb oligopeptide concentrate, enters degerming process to it.
12, degerming
After being filtered using molecular weight 60-80KDa hollow fiber membrane ultrafiltration devices, enters back into 0.22 μm of micropore filter element and be filtered, pressing
Under the conditions of power 2.0-3.0 MPa, ultrafiltration feed velocity 1.2-1.8L/min, sheep placenta lamb oligopeptide is concentrated by way of retention
Liquid carries out degerming, after degerming, is sealed into filling with stainless steel sealing bucket.
13, filling
Oligopeptide concentrate after degerming is carried out with automatic filling machine it is filling, using cleaning in sterile 10ml cillin bottles.West
Woods bottle first pass through ultraviolet light pass-through box enter ultrasonic bottle washing machine between wash bottle it is per minute wash 200 bottles of washed bottles and enter dryer exist
It is dried under 260 DEG C of 280 DEG C of preheating temperature, 300 DEG C of sterilising temp, heat preservation programs, completes 10,000 bottles of cillin bottle sterilizings per hour and dry
It is dry.Cillin bottle is filling into sterile beginning, and dosage controls within 5ml, ensures the stabilization of net content, per bottled amount 200mg,
Filling 200 bottles per minute, and after pressurized rubber plug to state of partly jumping a queue, canned product is entered in vacuum freeze drier
Freeze-drying.
14, it is freeze-dried
（1）Pre-freeze cures
By canned concentrate cillin bottle, it is placed on the partition board in vacuum freeze drier dry storehouse, pre-freeze solidification temperature -30
~-45 DEG C, and 5~8h is kept, so that moisture in concentrate is fully freezed, it is ensured that no liquid exists.
（2）Lyophilization：Freeze dryer starts vacuum system, and pressure in storehouse is evacuated to 30~50pa, takes slow heating, with
1 DEG C of heating rate per hour is slowly warming up to -20~-25 DEG C, keeps the temperature 6~8h, then with 2 DEG C per hour of heating rate, delay
It is slow to be warming up to -10 DEG C~-12 DEG C, 3~6h is kept the temperature, lyophilization is carried out.
（3）Parsing-desiccation：After lyophilization, then with 3 DEG C per hour of heating rate, 40~50 DEG C are slowly warming up to, pressure
Power is 10~20Pa.6~8h is kept the temperature, parsing-desiccation freeze-drying is completed, is taken out after the plastic lid that pressurizes.
15, bottle cap and plug, labelling are pressed
Aseptically machinery pressure bottle cap and plug after freeze-drying, makes it post label into labeling.Purity of protein is made to be up to
99.5%, oligopeptide component content of the relative molecular mass less than 1000Da is averagely more than 88% sheep placenta lamb oligopeptide finished powder.
Embodiment 2
It weighs 100kg and cleans the sheep placenta lamb that freezes, be put into after slicer is cut into the thin slice of 0.3-0.6cm thickness, inject pure water, soak
16-20h is steeped into doughy state.Put the fragment rubbed in meat grinder at granularity is 0.1-0.2cm.It is de- with 1.0-2.0% acetums
Fat 1-2d, with 30-50 DEG C of warm water rinse 2 times.Particle, which is worn into, with bone mud barreling refines slurries in 20~50 μm of sizes.Adjust pH value
4.0~4.5, NaCl powder is added and saltouts, with plate and frame filter press press filtration, transparent filtrate will be obtained and adjust pH value to be 9.0
~9.5, under the conditions of 50 DEG C~60 DEG C, 1-3% trypsin digestions are added and react 8h~10h.Enzymolysis liquid is classified ultrafiltration, first
By the ultrafilter of molecular weight 75KDa, filtered using the ultrafilter of molecular weight 3KDa.Its temperature is warming up to 80 after ultrafiltration~
95 DEG C of progress high temperature enzyme deactivations.Again through molecular weight be 500Da nanofiltration device in concentrated.Elution 3-5 times, makes to be concentrated into water content
The only minimum volume of 3-5% or so.The nanofiltration filtered fluid of impure and inorganic salts etc. is subjected to purified treatment.Oligopeptide is dense
After contracting liquid is using the filtering of molecular weight 60-80KDa hollow fiber membrane ultrafiltration devices, 0.22 μm of micropore filter element filtration sterilization is entered back into.With certainly
Dynamic bottle placer progress is filling, and 10ml cillin bottle dosage controls within 5ml, per bottled amount 200mg.Vacuum freeze drier freezes
It is dry.Purity of protein is made and is up to oligopeptide component content of the 99.5%, relative molecular mass less than 1000Da averagely up to 88%
Sheep placenta lamb oligopeptide finished powder.100kg, which freezes fresh sheep placenta lamb raw material, can averagely produce the oligomeric Gly-His-Lys of 3.6kg.
Claims (1)
1. a kind of industrialized producing technology of sheep placenta lamb albumen oligopeptide, its step are as follows：
（1）Raw material
Raw material selects sheep placenta lamb；
（2）Slice
It is cut into the thin slice of 0.3-0.6cm thickness；
（3）It impregnates and thaws
48h-72 h are impregnated in water filling, until sheep placenta lamb is at doughy state；
（4）It rubs
Rub into the fragment that granularity is 0.1-0.2cm；
（5）Degreasing
By the degreaser soak degreasing 1-2d of the sheep placenta lamb of rubbing 1.0-2.0% acetums, finally 30-50 DEG C of warm water is used to float
It washes 2 times, obtains the sheep placenta lamb that degreasing is impregnated；
（6）Defibrination
Particle is worn into 20~50 μm of grinding slurries；
（7）It saltouts
Grinding slurry obtained is adjusted into pH value 4.0~4.5, NaCl powder is added and saltouts, standing 12~for 24 hours, obtain collagen
Mixed liquor；
（8）Press filtration
Mixed liquor is uniformly routed on sheet frame filter cloth, conveying mixed liquor starts 1~6h of press filtration, discards filter cake, obtains transparent
Filtrate；
（9）Enzymolysis
Under the conditions of being 9.0~9.5,50 DEG C~60 DEG C by the slurries adjusting pH value after press filtration, 1-3% trypsase is then added,
So that sheep placenta lamb proteolysis is reacted 8h~10h, is obtained after being stirred continuously with 30 turns/min in enzymolysis process mixed containing oligopeptide
Close liquid；
（10）Ultrafiltration oligopeptide
Enzymolysis liquid is subjected to classification ultrafiltration through reverse osmosis ultrafiltration unit, the ultrafilter of molecular weight 75KDa is first passed through, using molecule
Measure the ultrafilter filtering of 3KDa；The peptide of suitable molecular weight segment enters collector through film, and the peptide of large fragment continues and enzyme effect
It digests again, then ultrafiltration obtains oligopeptide filtered fluid；Its temperature is warming up to 80~95 DEG C of progress high temperature enzyme deactivations after ultrafiltration；
（11）Nanofiltration retains concentration, desalination, removing impurities matter
It is concentrated in the nanofiltration device that the molecular weight that albumen oligopeptide filtrate is pumped into reverse osmosis nanofiltration unit is 500Da；Again will
Concentrate adds same volume water elution 3-5 times, and acquisition is concentrated into the minimum volume nanofiltration retentate fluid that water content only has 3-5%；It will
It collects containing other substances and the nanofiltrations filtered fluid such as inorganic salts in raw material, carries out purified treatment, the nanofiltration retentate fluid of collection is
Oligopeptide concentrate；
（12）Degerming
After concentrate is filtered using molecular weight 60-80KDa hollow fiber membrane ultrafiltration devices, enters back into 0.22 μm of micropore filter element and carried out
Filter, carries out degerming by way of retention；
（13）It is filling
With filling in cillin bottle；
（14）Freeze-drying
（a）Pre-freeze cures
By canned concentrate cillin bottle, it is placed on the partition board in vacuum freeze drier dry storehouse, pre-freeze solidification temperature -30
~-45 DEG C, and 5~8h is kept, so that moisture in concentrate is fully freezed, it is ensured that no liquid exists；
（b）Lyophilization：Freeze dryer starts vacuum system, and pressure in storehouse is evacuated to 30~50pa, takes slow heating, with every small
When 1 DEG C of heating rate, be slowly warming up to -20~-25 DEG C, keep the temperature 6~8h, then with 2 DEG C per hour of heating rate, slowly rise
Temperature arrives -10 DEG C~-12 DEG C, keeps the temperature 3~6h, carries out lyophilization；
（c）Parsing-desiccation：After lyophilization, then with 3 DEG C per hour of heating rate, 40~50 DEG C are slowly warming up to, pressure is
10~20Pa；6~8h is kept the temperature, parsing-desiccation freeze-drying is completed, is taken out after the plastic lid that pressurizes；
（15）Press bottle cap and plug, labelling
Aseptically machinery pressure bottle cap and plug after freeze-drying, makes it post label into labelling machine；Purity of protein height is made
Oligopeptide component content up to 99.5%, relative molecular mass less than 1000Da is averagely more than 88% sheep placenta lamb oligopeptide finished product
Powder.
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| CN1923214A (en) * | 2005-08-29 | 2007-03-07 | 包头蒙肽生物科技有限公司 | Preparation of sheep placenta peptide |
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| CN102329842A (en) * | 2011-07-12 | 2012-01-25 | 湖州康海斯生物科技有限公司 | Lamb placenta polypeptide and preparation method thereof |
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Application publication date: 20180814 |