CN108823270A - A kind of extracting method of oyster peptide - Google Patents
A kind of extracting method of oyster peptide Download PDFInfo
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- CN108823270A CN108823270A CN201810475406.3A CN201810475406A CN108823270A CN 108823270 A CN108823270 A CN 108823270A CN 201810475406 A CN201810475406 A CN 201810475406A CN 108823270 A CN108823270 A CN 108823270A
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- 241000237502 Ostreidae Species 0.000 title claims abstract description 161
- 235000020636 oyster Nutrition 0.000 title claims abstract description 161
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 91
- 238000000034 method Methods 0.000 title claims abstract description 25
- 235000013372 meat Nutrition 0.000 claims abstract description 51
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 claims abstract description 32
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 claims abstract description 32
- 239000004365 Protease Substances 0.000 claims abstract description 30
- 108091005804 Peptidases Proteins 0.000 claims abstract description 29
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract description 29
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 28
- 102000004190 Enzymes Human genes 0.000 claims abstract description 27
- 108090000790 Enzymes Proteins 0.000 claims abstract description 27
- 239000002781 deodorant agent Substances 0.000 claims abstract description 25
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 claims abstract description 23
- 150000001875 compounds Chemical class 0.000 claims abstract description 20
- 230000007071 enzymatic hydrolysis Effects 0.000 claims abstract description 12
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 claims abstract description 12
- WHGYBXFWUBPSRW-FOUAGVGXSA-N beta-cyclodextrin Chemical compound OC[C@H]([C@H]([C@@H]([C@H]1O)O)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O3)[C@H](O)[C@H]2O)CO)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H]3O[C@@H]1CO WHGYBXFWUBPSRW-FOUAGVGXSA-N 0.000 claims abstract description 8
- 229920000858 Cyclodextrin Polymers 0.000 claims abstract description 7
- 239000001116 FEMA 4028 Substances 0.000 claims abstract description 7
- 235000011175 beta-cyclodextrine Nutrition 0.000 claims abstract description 7
- 229960004853 betadex Drugs 0.000 claims abstract description 7
- 238000001914 filtration Methods 0.000 claims abstract description 7
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- 108090000623 proteins and genes Proteins 0.000 claims description 27
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- 238000000108 ultra-filtration Methods 0.000 claims description 12
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- 230000000996 additive effect Effects 0.000 claims description 11
- 238000001728 nano-filtration Methods 0.000 claims description 11
- SRBFZHDQGSBBOR-OWMBCFKOSA-N L-ribopyranose Chemical compound O[C@H]1COC(O)[C@@H](O)[C@H]1O SRBFZHDQGSBBOR-OWMBCFKOSA-N 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- RYYKJJJTJZKILX-UHFFFAOYSA-M sodium octadecanoate Chemical compound [Na+].CCCCCCCCCCCCCCCCCC([O-])=O RYYKJJJTJZKILX-UHFFFAOYSA-M 0.000 claims description 7
- JRMUNVKIHCOMHV-UHFFFAOYSA-M tetrabutylammonium bromide Chemical compound [Br-].CCCC[N+](CCCC)(CCCC)CCCC JRMUNVKIHCOMHV-UHFFFAOYSA-M 0.000 claims description 7
- 235000013601 eggs Nutrition 0.000 claims description 2
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 claims 1
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- 101000693530 Staphylococcus aureus Staphylokinase Proteins 0.000 description 6
- 244000005700 microbiome Species 0.000 description 6
- 235000020995 raw meat Nutrition 0.000 description 6
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- 241000251468 Actinopterygii Species 0.000 description 3
- 102000003886 Glycoproteins Human genes 0.000 description 3
- 108090000288 Glycoproteins Proteins 0.000 description 3
- 150000001299 aldehydes Chemical class 0.000 description 3
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- 238000006731 degradation reaction Methods 0.000 description 3
- 238000004332 deodorization Methods 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- DNJIEGIFACGWOD-UHFFFAOYSA-N ethanethiol Chemical compound CCS DNJIEGIFACGWOD-UHFFFAOYSA-N 0.000 description 3
- 239000000796 flavoring agent Substances 0.000 description 3
- 235000019634 flavors Nutrition 0.000 description 3
- 230000007062 hydrolysis Effects 0.000 description 3
- 238000006460 hydrolysis reaction Methods 0.000 description 3
- 108091008146 restriction endonucleases Proteins 0.000 description 3
- MMFCJPPRCYDLLZ-CMDGGOBGSA-N (2E)-dec-2-enal Chemical compound CCCCCCC\C=C\C=O MMFCJPPRCYDLLZ-CMDGGOBGSA-N 0.000 description 2
- QOPYYRPCXHTOQZ-CMDGGOBGSA-N (e)-dec-2-en-1-ol Chemical compound CCCCCCC\C=C\CO QOPYYRPCXHTOQZ-CMDGGOBGSA-N 0.000 description 2
- 102000007698 Alcohol dehydrogenase Human genes 0.000 description 2
- 108010021809 Alcohol dehydrogenase Proteins 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 102000008186 Collagen Human genes 0.000 description 2
- 108010035532 Collagen Proteins 0.000 description 2
- 241000548230 Crassostrea angulata Species 0.000 description 2
- 206010051820 Sordes Diseases 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 229920001436 collagen Polymers 0.000 description 2
- 239000002131 composite material Substances 0.000 description 2
- 238000010276 construction Methods 0.000 description 2
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- 230000004151 fermentation Effects 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 230000002209 hydrophobic effect Effects 0.000 description 2
- 150000002576 ketones Chemical class 0.000 description 2
- 238000003754 machining Methods 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 235000015170 shellfish Nutrition 0.000 description 2
- 230000002195 synergetic effect Effects 0.000 description 2
- 102000005369 Aldehyde Dehydrogenase Human genes 0.000 description 1
- 108020002663 Aldehyde Dehydrogenase Proteins 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- 241000237519 Bivalvia Species 0.000 description 1
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 1
- 241001522252 Crassostrea rivularis Species 0.000 description 1
- 244000086443 Craterellus fallax Species 0.000 description 1
- 235000007926 Craterellus fallax Nutrition 0.000 description 1
- WHGYBXFWUBPSRW-UHFFFAOYSA-N Cycloheptaamylose Natural products O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO WHGYBXFWUBPSRW-UHFFFAOYSA-N 0.000 description 1
- 108010002156 Depsipeptides Proteins 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 108090000270 Ficain Proteins 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 102000007079 Peptide Fragments Human genes 0.000 description 1
- 108010033276 Peptide Fragments Proteins 0.000 description 1
- 241000490567 Pinctada Species 0.000 description 1
- 102100025292 Stress-induced-phosphoprotein 1 Human genes 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 239000011149 active material Substances 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 238000013475 authorization Methods 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 238000006356 dehydrogenation reaction Methods 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 235000019836 ficin Nutrition 0.000 description 1
- POTUGHMKJGOKRI-UHFFFAOYSA-N ficin Chemical compound FI=CI=N POTUGHMKJGOKRI-UHFFFAOYSA-N 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 238000011017 operating method Methods 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000035911 sexual health Effects 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- RKHXQBLJXBGEKF-UHFFFAOYSA-M tetrabutylphosphanium;bromide Chemical compound [Br-].CCCC[P+](CCCC)(CCCC)CCCC RKHXQBLJXBGEKF-UHFFFAOYSA-M 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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- Meat, Egg Or Seafood Products (AREA)
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Abstract
The invention discloses a kind of extracting methods of oyster peptide, it is homogenized after oyster meat is removed internal organs, active dry yeast and arabinose deodorant is first added and then beta-cyclodextrin is added and is handled, compound protease progress constant temperature enzymatic hydrolysis is added after adjusting deodorant oyster meat homogenate pH value, enzyme deactivation after enzymatic hydrolysis, centrifuged supernatant filtering, is freeze-dried to get oyster peptide.It has the beneficial effect that：Low, easy to operate, protein degradation effect is good, oyster peptide yield is high, the higher advantage of sensory-acceptance with putting into for preparation method of the present invention, it is easy to industrial-scale production, the even molecular weight distribution of obtained oyster peptide, oyster peptide content of the molecular weight ranges in 4000-6000 is greater than 87%, has good application value and market potential.
Description
Technical field
The present invention relates to bioactive ingredients technical field, in particular to a kind of extracting method of oyster peptide.
Background technique
Oyster（oyster）It is to belong to Mollusca Bivalvia pearl shell purpose Qu Li section and Ostreidae（Ostreidae,
True oyster）Or Yan Ha section（Aviculidae, pearl oyster）Bivalve also known as oyster, oyster, oyster be yellow, oyster clam
Or male clam.Oyster is the first big cultivated shellfish in the world, worldwide it has been found that oyster kind have 100 kinds or so, it is male
Oyster is also China important marine shellfish, has more than 20 kinds in the oyster of China coast distribution, mainly has Crassostrea rivularis, Dalian Bay male
Oyster, Pacific oyster（Long oyster）（Crassostrea gigas）And close squama oyster.China is oyster culture big country, oyster money
Source electrode horn of plenty.In recent years, growth trend is presented in China oyster yield, however relies solely on conventional machining techniques production oyster and produce
Oneself is unable to satisfy the social development need to oyster industry now to product, and it is even more impossible to meet the majority of consumers to high-quality oyster product
Demand.Therefore using oyster as raw material, produced using oyster functional sexual health of the advanced machining technology production rich in various active substance
Product or drug have vast potential for future development, oneself becomes the hot spot studied both at home and abroad.Study oyster intensive processing and comprehensive utilization
Technology can not only promote the sound development of oyster secondary industry, while also can be the hair of China's modern times aquatic products intensive processing industry
Exhibition provides certain theoretical basis, and bigger economic benefit can be also created for enterprise.
The shell and software of oyster utility value all with higher, make full use of oyster software and oyster shell development function male
Oyster product has important social value and economic value.Protein content is up to 50%, and amino in oyster software dry matter
Acid composition is perfect, is good protein.But the isolation technics and process for mostly using tradition many and diverse during domestic production, make
The active component content of product is relatively low.To make full use of its nutritional ingredient and physiological activator, needing to release has
Some " inert proteins " are changed into " active peptide " by active albumen and peptide fragment.Mmp reaction mild condition will not destroy ammonia
The nutriments such as base acid, and reaction process is easily controllable, is used widely in oyster process.Oyster Protein matter is through egg
Many small molecule skins and amino acid are generated after white enzyme hydrolysis, not only contributes to absorption of human body, improve the utilization rate of protein, and
Active material can also be extracted.
The prior art such as Authorization Notice No. is the Chinese invention patent of CN104694606B, discloses a kind of complex enzyme water
The method that solution Oyster Protein prepares small-molecular peptides, specially：(1) oyster is cleaned, water and ficin is added, carry out enzyme
Solution, takes oyster meat；(2) oyster meat is smashed, water is added, stirs evenly, obtains oyster slurries；(3) complex enzyme, enzyme are added
Solution, enzyme deactivation obtain enzymolysis liquid；(4) enzymolysis liquid is subjected to ultrafiltration, is freeze-dried, obtains oyster small-molecular peptides.This method technique letter
It is single, it is easily operated, mechanical dejacketing is digested, the time is saved, shortens the production cycle, the protein peptides being prepared are small-molecular peptides,
Molecular weight is small, and activity is high.But the small-molecular peptides fishy smell is heavier, sensory-acceptance is poor, strongly limits such function factor
Application in health care product, medicine field.
Summary of the invention
The purpose of the present invention is to provide one kind, and with putting into, low, easy to operate, protein degradation effect is good, oyster peptide obtains
Rate height, the higher advantage of sensory-acceptance, the extracting method of the oyster peptide of the even molecular weight distribution of obtained oyster peptide.
The present invention in view of the above technology in the problem of mentioning, the technical solution taken is：
A kind of extracting method of oyster peptide, is homogenized after oyster meat is removed internal organs, and active dry yeast is first added and arabinose is de-
Raw meat, then addition beta-cyclodextrin is handled, and addition compound protease carries out constant temperature enzyme after adjusting deodorant oyster meat homogenate pH value
It solves, enzyme deactivation after enzymatic hydrolysis, centrifuged supernatant filtering is freeze-dried to get oyster peptide.The preparation method, which has, puts into low, behaviour
Work is simple, protein degradation effect is good, oyster peptide yield is high, the higher advantage of sensory-acceptance, is easy to industrial-scale production,
The even molecular weight distribution of obtained oyster peptide, oyster peptide content of the molecular weight ranges in 4000-6000 are greater than 87%, have good
Good application value and market potential.
Preferably, defishying the specific steps are：Active dry yeast and arabinose will be added in homogenate, in 38-
Then the beta-cyclodextrin of oyster meat weight 5-8% is added, temperature is 60-70 DEG C, super-pressure is in deodorant 20-30min at 43 DEG C
Super-pressure defishying 10-20min is under conditions of 300-400MPa to get the homogenate of deodorant oyster meat.Active dry yeast is loose
Structure has a suction-operated to stench substance, while active dry yeast contains there are many enzyme, first with active dry yeast by raw meat in the step
Sordes matter is converted into without substance of smelling as of rotten fish, then the macromolecular substances such as substance such as aldehyde, ketone of smelling as of rotten fish recycle β-ring-type by cell accumulation
The embedding effect of dextrin removes remaining yeast fermented flavour and fermentation acid and remaining fishy smell, reaches more preferably except raw meat effect, mentions
The sensory-acceptance of high target product.
Preferably, the additive amount of active dry yeast is oyster meat weight 0.8-1.3%, the additive amount of arabinose is male
Oyster meat weight 0.1-0.3%.The presence of arabinose can provide energy consumption for active dry yeast, improve alcohol dehydrogenase and aldehyde dehydrogenation
Fishy smell substance E-2- decenal is converted to E-2- decenol and E-2- decylenic acid, improves deodorization effect and mesh by the metabolism amount of enzyme
The quality of product is marked, while being avoided that active dry yeast consumes clam albumen, improves the yield of target product, and oyster can be made
The even molecular weight distribution of peptide.
Preferably, containing the D-arabinose of 4.3-4.6% in arabinose.The spy of D-arabinose in arabinose
It is different to exist, so that the albumen of different structure is very sensitive to pressure in oyster, the construction of Oyster Protein molecule is made to change,
It is denaturalized, so that internal restriction enzyme site exposure, is easy to by protease digestion, while oyster growing environment keeps itself contained micro-
Biomass is big, and mild condition is conducive to microbial growth during proteolysis, by the quality and yield production to product
Raw adverse effect, and D-arabinose cooperation super-pressure effect can change the Premeabilisation of cells pressure of microorganism, kill such micro- life
Object improves the yield and quality of oyster peptide.
Preferably, the weight ratio of oyster meat and water is 1 in homogenate:7-10.
Preferably, enzymatic hydrolysis the specific steps are：The pH to 7.5-8.5 that deodorant oyster meat is homogenized is adjusted, is by enzyme concentration
Compound protease is added in 3-4%, adds the sodium stearate of compound protease weight 2-2.6% and the tetrabutyl phosphonium bromide of 0.3-0.5%
Then ammonium digests 8-12h at 30-40 DEG C to get enzymolysis liquid.The special presence of sodium stearate and tetrabutylammonium bromide can not only
The hydrophobic region of Oyster Protein is enough acted on, dispersibility and stability of the Oyster Protein in oyster meat homogenate is improved, improves oyster
The touch opportunity of albumen and protease, and then enzymatic hydrolysis rate and oyster peptide yield are improved, and the glycosidic bond of glycoprotein can be cut,
Make glycogen degradation, so that protease quickly identifies that the albumen for the glycogen that is removed is digested, final raising oyster peptide
Yield, and the even molecular weight distribution of obtained oyster peptide, oyster peptide content of the molecular weight ranges in 4000-6000 are greater than 87%.
Preferably, compound protein is neutral proteinase and taste protease, weight ratio 1:2.3-2.7.Using neutrality
Protease and flavor protease composite hydrolysis can carry out digestion from different amino acid sites, so that it is more to generate a large amount of activity
Peptide, while leading to reinfocing effect due to synergistic function between enzyme, the polypeptide fragment function of accordingly generating can be further
Enhance, the content of polypeptide and the activity of active peptides in the final enzymolysis efficiency for improving albumen, enzymolysis liquid.
Preferably, filtering the specific steps are：Enzymolysis liquid is surpassed through the ultrafiltration membrane that molecular cut off is 8-12kDa
Filter obtains 8-12kDa peptide mixer below, then the nanofiltration membrane nanofiltration through 200-400Da, obtains the dense of 200-400Da or more
Depsipeptides liquid is freeze-dried up to oyster peptide.It is existing not to be fully hydrolyzed since the microorganism zymolyte comparison of ingredients of protein is complicated
Protein macromolecule, the peptide for also having molecular size range not equal and free amino acid, step separation belong to physics mode, do not have
There is introducing chemical reagent, preferably saves the primary characteristic of collagen peptide, and have to enzymolysis liquid and preferably isolate and purify effect
Fruit.
Compared with the prior art, the advantages of the present invention are as follows：1）Preparation method of the present invention have put into it is low, easy to operate,
Protein degradation effect is good, oyster peptide yield is high, the higher advantage of sensory-acceptance, is easy to industrial-scale production, obtains
The even molecular weight distribution of oyster peptide has good application value and market potential；2）The preparation method deodorization effect is excellent,
The sensory-acceptance of oyster peptide is improved, the restriction enzyme site exposure inside Oyster Protein can be made during deodorant, is easy to by protease
Digestion can be changed the Premeabilisation of cells pressure of microorganism, kill this quasi-microorganism, improve the yield and quality of oyster peptide；3）The preparation
Method can improve the touch opportunity of Oyster Protein and protease, can cut the glycosidic bond of glycoprotein, make glycogen degradation, improve enzymatic hydrolysis
Rate and oyster peptide yield, and obtain molecular weight ranges and be greater than 87% in the oyster peptide content of 4000-6000.
Specific embodiment
The present invention program is described further below by embodiment：
Embodiment 1：
A kind of extracting method of oyster peptide, is homogenized after oyster meat is removed internal organs, and active dry yeast is first added and arabinose is de-
Raw meat, then addition beta-cyclodextrin is handled, and addition compound protease carries out constant temperature enzyme after adjusting deodorant oyster meat homogenate pH value
It solves, enzyme deactivation after enzymatic hydrolysis, centrifuged supernatant filtering is freeze-dried to get oyster peptide.The preparation method, which has, puts into low, behaviour
Work is simple, protein degradation effect is good, oyster peptide yield is high, the higher advantage of sensory-acceptance, is easy to industrial-scale production,
The even molecular weight distribution of obtained oyster peptide, oyster peptide content of the molecular weight ranges in 4000-6000 are greater than 87%, have good
Good application value and market potential.
Defishying the specific steps are：Active dry yeast and arabinose, the deodorant at 38 DEG C will be added in homogenate
Then the beta-cyclodextrin of oyster meat weight 5%, superelevation under conditions of temperature is 70 DEG C, super-pressure is 300MPa is added in 30min
Press defishying 20min to get the homogenate of deodorant oyster meat.The loose structure of active dry yeast has suction-operated to stench substance,
Active dry yeast contains there are many enzyme simultaneously, is first converted stench substance to without substance of smelling as of rotten fish, raw meat with active dry yeast in the step
Then the macromolecular substances such as sordes matter such as aldehyde, ketone recycle the embedding effect of cycloheptaamylose to remove remaining by cell accumulation
Yeast fermented flavour and fermentation acid and remaining fishy smell, reach more preferably except raw meat effect, improve the sensory-acceptance of target product.
The additive amount of above-mentioned active dry yeast is oyster meat weight 0.8%, and the additive amount of arabinose is oyster meat weight
0.3%.The presence of arabinose can provide energy consumption for active dry yeast, improve the metabolism amount of alcohol dehydrogenase and aldehyde dehydrogenase, will
Fishy smell substance E-2- decenal is converted to E-2- decenol and E-2- decylenic acid, improves the quality of deodorization effect and target product,
It is avoided that active dry yeast consumes clam albumen simultaneously, improves the yield of target product, and the molecular weight point of oyster peptide can be made
Cloth is uniform.
In above-mentioned arabinose containing 4.3% D-arabinose.The special presence of D-arabinose, makes in arabinose
The albumen for obtaining different structure in oyster is very sensitive to pressure, so that the construction of Oyster Protein molecule is changed, is denaturalized,
So that internal restriction enzyme site exposure, is easy to by protease digestion, while oyster growing environment makes itself contained micro organism quantity
Greatly, condition mild during proteolysis is conducive to microbial growth, by the quality and the unfavorable shadow of yield generation to product
It rings, and D-arabinose cooperation super-pressure effect can change the Premeabilisation of cells pressure of microorganism, kill this quasi-microorganism, improve male
The yield and quality of oyster peptide.
The weight ratio of oyster meat and water is 1 in above-mentioned homogenate:10.
Enzymatic hydrolysis the specific steps are：The pH to 7.5 for adjusting the homogenate of deodorant oyster meat is 4% addition compound protein by enzyme concentration
Enzyme adds the sodium stearate of compound protease weight 2% and 0.5% tetrabutylammonium bromide, then digests 12h at 30 DEG C,
Up to enzymolysis liquid.The special presence of sodium stearate and tetrabutylammonium bromide can not only act on the hydrophobic region of Oyster Protein, mention
Dispersibility and stability of the high Oyster Protein in oyster meat homogenate, improve the touch opportunity of Oyster Protein and protease, in turn
Enzymatic hydrolysis rate and oyster peptide yield are improved, and the glycosidic bond of glycoprotein can be cut, makes glycogen degradation, so that protease is fast
Speed identifies that the albumen for the glycogen that is removed is digested, the final yield for improving oyster peptide, and the molecular weight of obtained oyster peptide
It is evenly distributed, oyster peptide content of the molecular weight ranges in 4000-6000 is greater than 87%.
Above-mentioned compound protein is neutral proteinase and taste protease, weight ratio 1:2.3.Using neutral proteinase and wind
Taste protease composite hydrolysis can carry out digestion from different amino acid sites, to generate a large amount of active peptides, while enzyme it
Between lead to reinfocing effect due to synergistic function, the polypeptide fragment function of accordingly generating can further enhance, and finally mention
The content of polypeptide and the activity of active peptides in the enzymolysis efficiency of high protein, enzymolysis liquid.
Filtering the specific steps are：Enzymolysis liquid is subjected to ultrafiltration through the ultrafiltration membrane that molecular cut off is 12kDa, is obtained
12kDa peptide mixer below, then the nanofiltration membrane nanofiltration through 200Da, obtain the concentration peptide liquid of 200Da or more, and freeze-drying is
Obtain oyster peptide.Since the microorganism zymolyte comparison of ingredients of protein is complicated, the existing protein macromolecule not being fully hydrolyzed,
There are molecular size range not equal peptide and free amino acid, step separation belongs to physics mode, does not introduce chemical reagent, compared with
The good primary characteristic for saving collagen peptide, and have to enzymolysis liquid and preferably isolate and purify effect.
Embodiment 2：
A kind of extracting method of oyster peptide, is homogenized after oyster meat is removed internal organs, and the weight ratio of oyster meat and water is 1 in homogenate:
8, active dry yeast and arabinose will be added in homogenate, wherein the additive amount of active dry yeast is oyster meat weight 1.0%, Ah
The additive amount for drawing uncle's sugar is oyster meat weight 0.2%, then β-ring paste of oyster meat weight 6% is added in the deodorant 25min at 40 DEG C
Essence, super-pressure defishying 15min obtains the homogenate of deodorant oyster meat under conditions of temperature is 65 DEG C, super-pressure is 350MPa, adjusts
The pH to 8.0 for saving the homogenate of deodorant oyster meat is 3.5% addition compound protease by enzyme concentration, adds compound protease weight
2.4% sodium stearate and 0.4% tetrabutylammonium bromide, then digest 10h at 35 DEG C and obtain enzymolysis liquid, then by enzymolysis liquid through cutting
It stays the ultrafiltration membrane that molecular weight is 10kDa to carry out ultrafiltration, obtains 10kDa peptide mixer below, then the nanofiltration membrane through 300Da is received
Filter obtains the concentration peptide liquid of 300Da or more, is freeze-dried up to oyster peptide, in above-mentioned arabinose containing 4.5% D- I
Uncle's sugar；Compound protein is neutral proteinase and taste protease, weight ratio 1:2.5.
Embodiment 3：
A kind of extracting method of oyster peptide, is homogenized after oyster meat is removed internal organs, and the weight ratio of oyster meat and water is 1 in homogenate:
10, active dry yeast and arabinose will be added in homogenate, wherein the additive amount of active dry yeast is oyster meat weight 0.8%, Ah
The additive amount for drawing uncle's sugar is oyster meat weight 0.3%, then β-ring paste of oyster meat weight 5% is added in the deodorant 30min at 38 DEG C
Essence, super-pressure defishying 20min obtains the homogenate of deodorant oyster meat under conditions of temperature is 70 DEG C, super-pressure is 300MPa, adjusts
The pH to 7.5 for saving the homogenate of deodorant oyster meat is 4% addition compound protease by enzyme concentration, adds compound protease weight 2%
Sodium stearate and 0.5% tetrabutylammonium bromide, then digest 12h at 30 DEG C and obtain enzymolysis liquid, then by enzymolysis liquid through retaining molecule
The ultrafiltration membrane that amount is 8kDa carries out ultrafiltration, obtains 8kDa peptide mixer below, then the nanofiltration membrane nanofiltration through 400Da, obtains
The concentration peptide liquid of 400Da or more is freeze-dried up to oyster peptide, in above-mentioned arabinose containing 4.3% D-arabinose；It is multiple
Hop protein is neutral proteinase and taste protease, weight ratio 1:2.7.
Comparative example 1：
A kind of extracting method of oyster peptide, is homogenized after oyster meat is removed internal organs, and the weight ratio of oyster meat and water is 1 in homogenate:
8, active dry yeast and arabinose will be added in homogenate, wherein the additive amount of active dry yeast is oyster meat weight 1.0%, Ah
The additive amount for drawing uncle's sugar is oyster meat weight 0.2%, then β-ring paste of oyster meat weight 6% is added in the deodorant 25min at 40 DEG C
Essence, super-pressure defishying 15min obtains the homogenate of deodorant oyster meat under conditions of temperature is 65 DEG C, super-pressure is 350MPa, adjusts
The pH to 8.0 for saving the homogenate of deodorant oyster meat is 3.5% addition compound protease by enzyme concentration, then digests 10h at 35 DEG C and obtain
Enzymolysis liquid, then enzymolysis liquid is subjected to ultrafiltration through the ultrafiltration membrane that molecular cut off is 10kDa, 10kDa peptide mixer below is obtained,
Nanofiltration membrane nanofiltration through 300Da again obtains the concentration peptide liquid of 300Da or more, is freeze-dried up to oyster peptide, above-mentioned arabinose
In containing 4.5% D-arabinose；Compound protein is neutral proteinase and taste protease, weight ratio 1:2.5.
In order to compare effect of the present invention, respectively to the molecule of oyster peptide prepared by embodiment 1, embodiment 2, comparative example 1
Amount is tested, concrete outcome such as table 1.
The molecular weight distribution table of 1 oyster peptide of table
As shown in Table 1, the oyster peptide obtained in embodiment 1, embodiment 2 and embodiment 3, molecular weight ranges are in 4000-6000
Content be greater than 87%, and comparative example 1 obtain oyster peptide molecular weight ranges 4000-6000 content be 68.4, illustrate ten
The even molecular weight distribution for the oyster peptide that the special presence of eight sour sodium and tetrabutylammonium bromide can make.
Routine operation in operating procedure of the invention is well known to those skilled in the art, herein without repeating.
Technical solution of the present invention is described in detail in embodiment described above, it should be understood that the above is only
For specific embodiments of the present invention, it is not intended to restrict the invention, all any modifications made in spirit of the invention,
Supplement or similar fashion substitution etc., should all be included in the protection scope of the present invention.
Claims (8)
1. a kind of extracting method of oyster peptide, it is characterised in that：It is homogenized after oyster meat is removed internal organs, active dry yeast is first added
With arabinose deodorant, then addition beta-cyclodextrin is handled, and compound protein is added after adjusting deodorant oyster meat homogenate pH value
Enzyme carries out constant temperature enzymatic hydrolysis, enzyme deactivation after enzymatic hydrolysis, and centrifuged supernatant filtering is freeze-dried to get oyster peptide.
2. a kind of extracting method of oyster peptide according to claim 1, it is characterised in that：The specific step of the defishying
Suddenly it is：Active dry yeast will be added in being homogenized and then oyster meat is added in arabinose, the deodorant 20-30min at 38-43 DEG C
The beta-cyclodextrin of weight 5-8%, super-pressure defishying under conditions of temperature is 60-70 DEG C, super-pressure is 300-400MPa
10-20min is to get the homogenate of deodorant oyster meat.
3. a kind of extracting method of oyster peptide according to claim 1, it is characterised in that：The addition of the active dry yeast
Amount is oyster meat weight 0.8-1.3%, and the additive amount of the arabinose is oyster meat weight 0.1-0.3%.
4. a kind of extracting method of oyster peptide according to claim 1, it is characterised in that：Contain in the arabinose
The D-arabinose of 4.3-4.6%.
5. a kind of extracting method of oyster peptide according to claim 1, it is characterised in that：Oyster meat and water in the homogenate
Weight ratio be 1:7-10.
6. a kind of extracting method of oyster peptide according to claim 1, it is characterised in that：The specific steps of the enzymatic hydrolysis
For：The pH to 7.5-8.5 for adjusting the homogenate of deodorant oyster meat is that compound protease is added in 3-4% by enzyme concentration, adds compound egg
The sodium stearate of white enzyme weight 2-2.6% and the tetrabutylammonium bromide of 0.3-0.5%, then digest 8-12h, i.e., at 30-40 DEG C
Obtain enzymolysis liquid.
7. a kind of extracting method of oyster peptide according to claim 1, it is characterised in that：The compound protein is neutral egg
White enzyme and taste protease, weight ratio 1:2.3-2.7.
8. a kind of extracting method of oyster peptide according to claim 1, it is characterised in that：The specific steps of the filtering
For：Enzymolysis liquid is subjected to ultrafiltration through the ultrafiltration membrane that molecular cut off is 8-12kDa, obtains 8-12kDa peptide mixer below, then
Nanofiltration membrane nanofiltration through 400-600Da obtains the concentration peptide liquid of 400-600Da or more, is freeze-dried up to oyster peptide.
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| CN111334549A (en) * | 2020-02-21 | 2020-06-26 | 海南美肽生物科技有限公司 | Oyster peptide and oyster peptide extraction method |
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