CN108740280A - The preparation method and its usage of scorpion activated protein - Google Patents
The preparation method and its usage of scorpion activated protein Download PDFInfo
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- 241000239226 Scorpiones Species 0.000 title claims abstract description 127
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 115
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 115
- 238000002360 preparation method Methods 0.000 title claims abstract description 22
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 60
- 239000002781 deodorant agent Substances 0.000 claims abstract description 22
- 238000005238 degreasing Methods 0.000 claims abstract description 21
- 238000000502 dialysis Methods 0.000 claims abstract description 20
- 239000003963 antioxidant agent Substances 0.000 claims abstract description 17
- 230000003078 antioxidant effect Effects 0.000 claims abstract description 17
- 239000000843 powder Substances 0.000 claims abstract description 17
- 235000006708 antioxidants Nutrition 0.000 claims abstract description 14
- 235000013372 meat Nutrition 0.000 claims abstract description 12
- 238000000926 separation method Methods 0.000 claims abstract description 11
- 235000020183 skimmed milk Nutrition 0.000 claims abstract description 9
- 235000019441 ethanol Nutrition 0.000 claims description 23
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 18
- 239000012153 distilled water Substances 0.000 claims description 14
- 238000001556 precipitation Methods 0.000 claims description 14
- 239000013049 sediment Substances 0.000 claims description 13
- 239000000284 extract Substances 0.000 claims description 10
- 238000004108 freeze drying Methods 0.000 claims description 10
- 238000000605 extraction Methods 0.000 claims description 9
- 239000006228 supernatant Substances 0.000 claims description 9
- 238000013019 agitation Methods 0.000 claims description 8
- 239000006227 byproduct Substances 0.000 claims description 8
- MWIXENPCUPDSOS-QMMMGPOBSA-N (2s)-2-amino-3-(4-hydroxyphenyl)propanehydrazide Chemical class NNC(=O)[C@@H](N)CC1=CC=C(O)C=C1 MWIXENPCUPDSOS-QMMMGPOBSA-N 0.000 claims description 7
- 238000007710 freezing Methods 0.000 claims description 6
- 230000008014 freezing Effects 0.000 claims description 6
- 235000007164 Oryza sativa Nutrition 0.000 claims description 5
- 238000001035 drying Methods 0.000 claims description 5
- 239000000835 fiber Substances 0.000 claims description 5
- 238000000265 homogenisation Methods 0.000 claims description 5
- 230000001376 precipitating effect Effects 0.000 claims description 5
- 235000009566 rice Nutrition 0.000 claims description 5
- 230000001476 alcoholic effect Effects 0.000 claims description 4
- 238000003860 storage Methods 0.000 claims description 4
- VKPPFDPXZWFDFA-UHFFFAOYSA-N 2-chloroethanamine Chemical compound NCCCl VKPPFDPXZWFDFA-UHFFFAOYSA-N 0.000 claims description 3
- 239000000463 material Substances 0.000 claims description 3
- 240000007594 Oryza sativa Species 0.000 claims 1
- 238000004140 cleaning Methods 0.000 claims 1
- 239000002244 precipitate Substances 0.000 claims 1
- 230000001681 protective effect Effects 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 20
- 239000002585 base Substances 0.000 abstract description 15
- 239000003513 alkali Substances 0.000 abstract description 13
- 239000012467 final product Substances 0.000 abstract description 9
- 238000004332 deodorization Methods 0.000 abstract description 4
- 238000000034 method Methods 0.000 abstract description 4
- 238000010525 oxidative degradation reaction Methods 0.000 abstract description 4
- 239000000047 product Substances 0.000 abstract description 4
- 239000002253 acid Substances 0.000 abstract 1
- 230000009286 beneficial effect Effects 0.000 abstract 1
- 235000015110 jellies Nutrition 0.000 abstract 1
- 239000008274 jelly Substances 0.000 abstract 1
- 239000000126 substance Substances 0.000 description 30
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 24
- 239000000243 solution Substances 0.000 description 20
- 150000001299 aldehydes Chemical class 0.000 description 14
- 150000001412 amines Chemical class 0.000 description 14
- 239000002956 ash Substances 0.000 description 10
- 239000007788 liquid Substances 0.000 description 9
- ONRREFWJTRBDRA-UHFFFAOYSA-N 2-chloroethanamine;hydron;chloride Chemical compound [Cl-].[NH3+]CCCl ONRREFWJTRBDRA-UHFFFAOYSA-N 0.000 description 8
- 235000002918 Fraxinus excelsior Nutrition 0.000 description 8
- 230000015556 catabolic process Effects 0.000 description 8
- 238000006731 degradation reaction Methods 0.000 description 8
- 239000012071 phase Substances 0.000 description 8
- -1 Quinoline ketone Chemical class 0.000 description 7
- 239000003814 drug Substances 0.000 description 7
- 239000012074 organic phase Substances 0.000 description 7
- 102000004190 Enzymes Human genes 0.000 description 6
- 108090000790 Enzymes Proteins 0.000 description 6
- 241000522620 Scorpio Species 0.000 description 6
- 150000002632 lipids Chemical class 0.000 description 6
- 239000009490 scorpio Substances 0.000 description 6
- 235000013305 food Nutrition 0.000 description 5
- 238000005259 measurement Methods 0.000 description 5
- 230000002195 synergetic effect Effects 0.000 description 5
- 241000209094 Oryza Species 0.000 description 4
- 229910052760 oxygen Inorganic materials 0.000 description 4
- 239000001301 oxygen Substances 0.000 description 4
- 238000004321 preservation Methods 0.000 description 4
- 238000003916 acid precipitation Methods 0.000 description 3
- 230000003712 anti-aging effect Effects 0.000 description 3
- 230000000844 anti-bacterial effect Effects 0.000 description 3
- 238000006701 autoxidation reaction Methods 0.000 description 3
- 210000000170 cell membrane Anatomy 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 235000019784 crude fat Nutrition 0.000 description 3
- 238000006481 deamination reaction Methods 0.000 description 3
- 238000006114 decarboxylation reaction Methods 0.000 description 3
- 230000002708 enhancing effect Effects 0.000 description 3
- 230000014759 maintenance of location Effects 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 241001481692 Mesobuthus martensii Species 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 210000000080 chela (arthropods) Anatomy 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 230000003859 lipid peroxidation Effects 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 239000006187 pill Substances 0.000 description 2
- 241000208340 Araliaceae Species 0.000 description 1
- 241000239290 Araneae Species 0.000 description 1
- 241000238421 Arthropoda Species 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 241000040710 Chela Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241001481690 Mesobuthus eupeus Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 1
- 235000003140 Panax quinquefolius Nutrition 0.000 description 1
- 206010047115 Vasculitis Diseases 0.000 description 1
- SMWDFEZZVXVKRB-UHFFFAOYSA-N anhydrous quinoline Natural products N1=CC=CC2=CC=CC=C21 SMWDFEZZVXVKRB-UHFFFAOYSA-N 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 230000003073 embolic effect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- 235000008434 ginseng Nutrition 0.000 description 1
- 150000002576 ketones Chemical group 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 238000011017 operating method Methods 0.000 description 1
- 238000005502 peroxidation Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000012460 protein solution Substances 0.000 description 1
- 150000003254 radicals Chemical class 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000002795 scorpion venom Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 206010044652 trigeminal neuralgia Diseases 0.000 description 1
- 210000004885 white matter Anatomy 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/02—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from meat
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Abstract
The invention discloses the preparation method and its usage of scorpion activated protein, the preparation method of scorpion activated protein, including：Degreasing, separation, dialysis, deodorant, it is heavy to acid after skimmed milk solution alkali soluble to obtain scorpion protein isolate first by scorpion meat jelly dry powder degreasing, then carry out dialysis and remove ash content, the ethanol solution deodorant containing 3- hydroxy-5-methyl base -2,4- imidazolones is added up to scorpion active protein powder.It has the beneficial effect that：Preparation method is simple, environment-friendly high-efficiency by the present invention, preparation method can efficiently act on the quaternary structure of protein, and the content of scorpion activated protein in final product can be greatly improved while avoiding protein by oxidative degradation, the method of the present invention is preferable to the deodorization effect of activated protein, and scorpion activated protein can be applied to anti-oxidant green nourishing product.
Description
Technical field
The present invention relates to the deep processing application fields of scorpion, more particularly, to the preparation method and its use of scorpion activated protein
On the way.
Technical background
Scorpion is the general designation of animal kingdom's Arthropoda Arachnoidea Scorpionida type, and spider also belongs to Arachnoidea.They are typical
Feature include tall and thin body, chela, curved section and the tail with telson（Postabdomen）.The earliest scorpion in land is about
Come across the uncommon retained profits subperiod (Silurian Period) before 400,013,000 years.There are about more than 1700 to plant for scorpion in the world, commonly use to be used as medicine,
It is buthus martensii Karscs to steep in wineKarsch, there is preferable therapeutic effect, also known as geneva just to clamp scorpion rheumatism, belong to the pincers of Scorpionida
Scorpion sectionButhidae.Buthus martensii Karscs quantity is most, and distribution is most wide, spreads all over China more than 10 and saves.Other scorpions of being used as medicine：Tibet plucked string instrument with a fretted fingerboard scorpionScorpiops Tibetanus；Other scorpions of steeping in wine：Streak clamps scorpion（Mongolian positive pincers scorpion）Mesobuthus Eupeus.The medicine of scorpio
Very high with being worth, the prescriptions of traditional Chinese medicine being made into scorpio is up to more than 100 kind.Scorpio be ginseng restorative pill, dahuoluo pills, it is seven precious red,
The important source material of more than the 30 kinds of Chinese patent drugs such as security personnel's cure-all medicine, Qianzheng San, is the common animal drug of China's tcm clinical practice.Especially
In recent years, scorpio had notable curative effect in the discovery on disease that cures difficult and complicated cases, as scorpio can treat vasculitis, embolic occlusion, scorpion venom
Cardiovascular disease, various tumours, trigeminal neuralgia etc. can be treated.These applications make the demand of scorpion sharply increase.In addition to medicinal,
Nourishing food can also be made in scorpio.With the progress of social material civilization, scorpion is as treatment, health-care good product, by people
Concern.
Invention content
The purpose of the present invention is to provide the preparation method and its usage of scorpion activated protein, preparation method of the invention letter
Single easy, environment-friendly high-efficiency, preparation method can efficiently act on the quaternary structure of protein, and avoid protein by oxygen
The content of scorpion activated protein in final product, deodorant of the method for the present invention to activated protein can be greatly improved by changing while degradation
Effect is preferable, and scorpion activated protein can be applied to anti-oxidant green nourishing product.
The present invention is directed to the problem of being mentioned in background technology, and the technical solution taken is：
The preparation method of scorpion activated protein, including：Degreasing, separation, dialysis, deodorant, specifically include following steps：
Degreasing：It is crushed after scorpion meat is cleaned up, is transferred in vacuum freeze drier, freezes after being placed in -80~-90 DEG C of freezings
Dry about 24~36h, crushes, using supercritical CO2It extracts degreasing and obtains fat-soluble by-product, the degreasing dregs of rice crushed 40~80
Mesh sieves, -20~-30 DEG C of storages；With supercritical CO2Scorpion meat degreasing is extracted, not only easy to operate, extraction efficiency is high, Er Qieke
To recycle fat-soluble by-product, realize that the full effect of scorpion meat utilizes；
Separation：Scorpion skimmed milk is weighed in container, according to solid-liquid ratio 1g:Distilled water is added in 20~25mL, molten with sodium hydroxide
Liquid adjusts pH to 11.5~11.8, and magnetic agitation extracts 50~60min under the conditions of 38~40 DEG C, then with 4000~4800r/
Min, 10~12min is centrifuged under the conditions of 1~4 DEG C, collect supernatant；The precipitating proteins of pH to 4.5~4.55 of supernatant are adjusted,
Then to centrifuge 8~10min under the conditions of 10000~12000r/min, 1~4 DEG C, sediment is collected, precipitation is washed with distilled water
Object 2~3 times, freeze-drying, -20~-25 DEG C of preservations obtain scorpion protein isolate；Under alkaline environment, in scorpion skimmed milk
Alkali-soluble reactive protein can be slowly dissolved in lye, protein can be precipitated through isoelectric precipitation, but with solution alkaline
Enhancing, scorpion albumen occur decarboxylation deamination reaction and generate bitter substance and other smell substances, lead to the peculiar smell of protein
Enhancing, color burn；
Dialysis：It is dialysed to scorpion separated protein powder using 12ku aperture fibers bag filter, solid-liquid ratio 1:200~230, temperature
Degree is 1~2 DEG C, and dialyse 48~72h, per a dialyzate is changed for 24 hours, collects precipitation, is preserved with -20~-25 DEG C after freeze-drying；
Low temperature dialysis can effectively reduce the content of ashes in scorpion protein isolate, and content of ashes can be reduced to 3% to divide hereinafter, increasing
Purity from protein in albumen and content；
Deodorant：Weigh the scorpion separated protein powder after dialysis, 4~10 times 95~98% of ethanol solution be added, 18~20 DEG C,
5~8min of high speed homogenization under the conditions of 10000~12500r/min, then with 1500~2000r/min rotating speeds centrifugation 10~
15min collects precipitation, and repeating deodorant with above-mentioned condition is placed at aeration-drying for 2~5 times, and sediment is allowed to be done naturally at room temperature
Dry scorpion active protein powder to obtain the final product；Some endogenous enzymes, microorganism in scorpion body etc. to the conversion of scorpion albumen, degradation and
After some aldehyde, the amine substance of lipid autoxidation generation can have certain bad smell, alkali extraction-acid precipitation to extract scorpion albumen,
Since the effect of alkali causes the scorpion albumen of extraction to generate certain alkali taste, scorpion activated protein can be influenced after being mixed with bad smell
Utilization；Water phase and organic phase in ethanol solution constitute two-phase system, can effectively remove with the fat-soluble of fishy smell
With water-soluble substances, the separation of fishy smell substance is realized, while ethyl alcohol is easier to remove by volatilizing.
Preferably, the 2-chloroethyl amine hydrochloric acid containing 2~3% ethyl alcohol, 0.5~0.6% in distilled water in separating step
Salt and 0.02~0.03 ‰ l-tyrosine hydrazides；The 2-chloroethyl amine hydrochloride of micro and special proportioning exists with l-tyrosine hydrazides
The quaternary structure that protein can be acted under alcoholic environment weakens the space arrangement mode of quaternary structure, is soaked in magnetic agitation
It carries under effect, the alkali solubility albumen in scorpion albumen can dissociate and enter in lye, while 2-chloroethyl amine hydrochloride can seize body
Oxide in system, avoids oxidative degradation of the oxide to protein, on the one hand avoids protein and is degraded to bad smell object
Matter reduces the fishy smell of activated protein, is easy to deodorant, on the other hand improves the content of scorpion activated protein in final product, improves
The biological activity of scorpion activated protein.
Preferably, containing 0.2~0.5% 3- hydroxy-5-methyl base -2,4- imidazoles in ethanol solution in deodorant step
Quinoline ketone, the content containing (R) -3- hydroxy-5-methyl base -2,4- imidazolones is 78 in 3- hydroxy-5-methyl base -2,4- imidazolones
~79%；Aldehyde, amine substance in scorpion activated protein system have certain oxidisability, may be made to scorpion activated protein
At certain degradation, and in ethanol solution specific proportioning (R) -3- hydroxy-5-methyl base -2,4- imidazolones Yu (S) -3- hydroxyls
There is base -5- methyl -2,4- imidazolones synergistic effect, the synergistic effect can greatly enhance aldehyde, amine substance in system
On the one hand dissolubility in organic phase can be rapidly separated out aldehyde, amine substance, improve deodorization effect, on the other hand also drop
The low degradation of aldehyde, amine substance to scorpion activated protein, improves the content and purity of scorpion activated protein in final product.
The purposes of scorpion activated protein, purposes of the scorpion activated protein in anti-oxidant nutriment, scorpion activated protein can
Body total antioxidant capacity, activities of antioxidant enzymes and GSH contents are effectively improved, to reduce mistake of the oxygen radical to Cell membrane lipids
Oxidation reduces body MDA contents, shows inhibiting effect to liver spontaneity lipid peroxidation, therefore certainly with removing
By biological effectiveness such as base, antibacterial, anti-oxidant and anti-agings, a kind of environmentally friendly, efficient anti-oxidant green can be applied to
Nutriment.
Compared with the prior art, the advantages of the present invention are as follows：
1）Water phase and organic phase in ethanol solution constitute two-phase system, can effectively remove with fishy smell it is fat-soluble with
Water-soluble substances realize the separation of fishy smell substance, while ethyl alcohol is easier to remove by volatilizing；
2）2-chloroethyl amine hydrochloride in separating step can avoid oxidative degradation of the oxide to protein significantly, both avoid
Protein is degraded to bad smell substance, reduces the fishy smell of activated protein, is easy to deodorant, and improves scorpion activity in final product
The content of albumen improves the biological activity of scorpion activated protein；
3）In separating step, the 2-chloroethyl amine hydrochloride of micro and special proportioning and l-tyrosine hydrazides can be under alcoholic environment
The quaternary structure for acting on protein weakens the space arrangement mode of quaternary structure, under magnetic agitation extraction effect, scorpion egg
Alkali solubility albumen in white can dissociate and enter in lye；
4）(R) -3- hydroxy-5-methyls base -2,4- imidazolones of specific proportioning and (S) -3- hydroxy-5-methyls base-in deodorant step
The synergistic effect of 2,4- imidazolones can greatly enhance the dissolubility of aldehyde, amine substance in organic phase in system, can be with
It is rapidly separated out aldehyde, amine substance, improves deodorization effect, while also reducing aldehyde, amine substance to the drop of scorpion activated protein
Solution, improves the content and purity of scorpion activated protein in final product；
5）Scorpion activated protein can effectively improve body total antioxidant capacity, activities of antioxidant enzymes and GSH contents, reduce oxygen radical
To the peroxidation of Cell membrane lipids, therefore has and removes the biological effectiveness such as free radical, antibacterial, anti-oxidant and anti-aging,
Environmentally friendly, efficient anti-oxidant green nourishing product can be applied to.
Specific implementation mode
The present invention program is described further below by embodiment：
Embodiment 1：
The preparation method of scorpion activated protein, includes the following steps：
1）It crushes, is transferred in vacuum freeze drier after being placed in -80 DEG C of freezings, freeze-drying is about after scorpion meat is cleaned up
For 24 hours, it crushes, using supercritical CO2It extracts degreasing and obtains fat-soluble by-product, the degreasing dregs of rice be crushed into 40 mesh sieve, -20 DEG C of storages
It hides；2）Scorpion skimmed milk is weighed in container, according to solid-liquid ratio 1g:Distilled water is added in 20mL, and pH is adjusted with sodium hydroxide solution
To 11.5, magnetic agitation extracts 50min under the conditions of 38 DEG C, then to centrifuge 10min under the conditions of 4000r/min, 1 DEG C, collects
Supernatant；The precipitating proteins of pH to 4.5 for adjusting supernatant are collected then to centrifuge 8min under the conditions of 10000r/min, 1 DEG C
Sediment, is washed with distilled water sediment 2 times, freeze-drying, and -20 DEG C of preservations obtain scorpion protein isolate；3）Using the holes 12ku
Diameter fiber dialysis bag dialyses to scorpion separated protein powder, solid-liquid ratio 1:200, temperature is 1 DEG C, and dialyse 48h, every to change for 24 hours
Dialyzate collects precipitation, is preserved with -20 DEG C after freeze-drying；4）The scorpion separated protein powder after dialysis is weighed, is added 4
Then 95% ethanol solution again, high speed homogenization 5min under the conditions of 18 DEG C, 10000r/min are centrifuged with 1500r/min rotating speeds
10min collects precipitation, and repeating deodorant with above-mentioned condition is placed at aeration-drying for 2 times, and sediment is allowed to spontaneously dry i.e. at room temperature
Obtain scorpion active protein powder.
Embodiment 2：
The preparation method of scorpion activated protein, includes the following steps：
1）Degreasing：It crushes, is transferred in vacuum freeze drier after being placed in -90 DEG C of freezings, freezing is dry after scorpion meat is cleaned up
Dry about 36h is crushed, using supercritical CO2It extracts degreasing and obtains fat-soluble by-product, the degreasing dregs of rice, which crushed 80 mesh, to be sieved, -30 DEG C
Storage；With supercritical CO2Scorpion meat degreasing is extracted, not only easy to operate, extraction efficiency is high, and can recycle fat-soluble by-product
Object realizes that the full effect of scorpion meat utilizes；
2）Separation：Scorpion skimmed milk is weighed in container, according to solid-liquid ratio 1g:Distilled water is added in 25mL, contains 3% in distilled water
Ethyl alcohol, 0.6% 2-chloroethyl amine hydrochloride and 0.03 ‰ l-tyrosine hydrazides；PH to 11.8 is adjusted with sodium hydroxide solution,
Magnetic agitation extracts 60min under the conditions of 40 DEG C, then to centrifuge 12min under the conditions of 4800r/min, 4 DEG C, collects supernatant；
The precipitating proteins of pH to 4.55 of supernatant are adjusted, then to centrifuge 10min under the conditions of 12000r/min, 4 DEG C, collect precipitation
Object, is washed with distilled water sediment 3 times, freeze-drying, and -25 DEG C of preservations obtain scorpion protein isolate；Under alkaline environment, scorpion
Alkali-soluble reactive protein in sub- skimmed milk can be slowly dissolved in lye, and protein can be precipitated through isoelectric precipitation, but
As solution alkaline enhances, scorpion albumen occurs decarboxylation deamination reaction and generates bitter substance and other smell substances, causes
The peculiar smell enhancing of protein, color burn；
3）Dialysis：It is dialysed to scorpion separated protein powder using 12ku aperture fibers bag filter, solid-liquid ratio 1:230, temperature
It it is 2 DEG C, dialyse 72h, per a dialyzate is changed for 24 hours, collects precipitation, is preserved with -25 DEG C after freeze-drying；Low temperature dialysis can have
Effect reduce scorpion protein isolate in content of ashes, content of ashes can be reduced to 3% hereinafter, increase protein isolate in protein
Purity and content；
4）Deodorant：The scorpion separated protein powder after dialysis is weighed, 10 times 98% of ethanol solution is added, contains in ethanol solution
0.5% 3- hydroxy-5-methyl base -2,4- imidazolones contain (R) -3- hydroxyls-in 3- hydroxy-5-methyl base -2,4- imidazolones
The content of 5- methyl -2,4- imidazolones is 79%；High speed homogenization 8min under the conditions of 20 DEG C, 12500r/min, then with
2000r/min rotating speeds centrifuge 15min, collect precipitation, and repeating deodorant with above-mentioned condition is placed at aeration-drying for 5 times, at room temperature
Sediment is allowed to spontaneously dry up to scorpion active protein powder；Some endogenous enzymes, microorganism in scorpion body etc. are to scorpion albumen
Conversion, some aldehyde of degradation and lipid autoxidation generation, amine substance can have certain bad smell, alkali extraction-acid precipitation
After extracting scorpion albumen, since the effect of alkali causes the scorpion albumen of extraction to generate certain alkali taste, meeting after being mixed with bad smell
Influence the utilization of scorpion activated protein；Water phase and organic phase in ethanol solution constitute two-phase system, can effectively remove
Fat-soluble and water-soluble substances with fishy smell realize the separation of fishy smell substance, while ethyl alcohol is easier to remove by volatilizing.
Embodiment 3：
The preparation method of scorpion activated protein, including：Degreasing, separation, dialysis, deodorant, specifically include following steps：
Degreasing：It is crushed after scorpion meat is cleaned up, is transferred in vacuum freeze drier, is freeze-dried after being placed in -90 DEG C of freezings
About 30h is crushed, using supercritical CO2It extracts degreasing and obtains fat-soluble by-product, the degreasing dregs of rice be crushed into 60 mesh sieve, -25 DEG C of storages
It hides；With supercritical CO2Scorpion meat degreasing is extracted, not only easy to operate, extraction efficiency is high, and can recycle fat-soluble by-product
Object realizes that the full effect of scorpion meat utilizes；
Separation：Scorpion skimmed milk is weighed in container, according to solid-liquid ratio 1g:Distilled water is added in 24mL, with sodium hydroxide solution tune
PH to 11.6 is saved, magnetic agitation extracts 60min under the conditions of 38 DEG C, then to centrifuge 10min under the conditions of 4500r/min, 2 DEG C,
Collect supernatant；The precipitating proteins of pH to 4.52 for adjusting supernatant, then to be centrifuged under the conditions of 10500r/min, 2 DEG C
10min collects sediment, is washed with distilled water sediment 3 times, is freeze-dried, and -22 DEG C of preservations obtain scorpion protein isolate；?
Under alkaline environment, the alkali-soluble reactive protein in scorpion skimmed milk can be slowly dissolved in lye, through isoelectric precipitation
Protein is precipitated, but as solution alkaline enhances, scorpion albumen occur decarboxylation deamination reaction generate bitter substance and other
Smell substance causes the peculiar smell of protein to enhance, color burn；
Dialysis：It is dialysed to scorpion separated protein powder using 12ku aperture fibers bag filter, solid-liquid ratio 1:210, temperature 1
DEG C, dialyse 72h, per a dialyzate is changed for 24 hours, collects precipitation, is preserved with -20 DEG C after freeze-drying；Low temperature dialysis can be effective
Reduce the content of ashes in scorpion protein isolate, content of ashes can be reduced to 3% hereinafter, increasing protein in protein isolate
Purity and content；
Deodorant：The scorpion separated protein powder after dialysis is weighed, 8 times 98% of ethanol solution is added, in 20 DEG C, 11000r/min items
Then high speed homogenization 6min under part centrifuges 12min with 1800r/min rotating speeds, collect precipitation, and deodorant 4 times is repeated with above-mentioned condition
It is placed at aeration-drying, allows sediment to spontaneously dry at room temperature up to scorpion active protein powder；Some in scorpion body are endogenous
Some aldehyde that enzyme, microorganism etc. generate the conversion of scorpion albumen, degradation and lipid autoxidation, amine substance can have
Certain bad smell, after alkali extraction-acid precipitation extracts scorpion albumen, since the effect of alkali causes the scorpion albumen of extraction to generate centainly
Alkali taste, the utilization of scorpion activated protein can be influenced after being mixed with bad smell；Water phase and organic phase in ethanol solution constitute
Two-phase system can effectively remove the fat-soluble and water-soluble substances with fishy smell, realize the separation of fishy smell substance, while second
Alcohol is easier to remove by volatilizing.
Contain 3% ethyl alcohol, 0.55% 2-chloroethyl amine hydrochloride and 0.02 ‰ L- junket in distilled water in separating step
Propylhomoserin hydrazides；The 2-chloroethyl amine hydrochloride of micro and special proportioning can act on egg with l-tyrosine hydrazides under alcoholic environment
The quaternary structure of white matter weakens the space arrangement mode of quaternary structure, under magnetic agitation extraction effect, the alkali in scorpion albumen
Dissolubility albumen can dissociate and enter in lye, while 2-chloroethyl amine hydrochloride can seize the oxide in system, avoid oxide
On the one hand oxidative degradation to protein avoids protein and is degraded to bad smell substance, reduces the fishy smell of activated protein, easily
In deodorant, the content of scorpion activated protein in final product is on the other hand improved, the biology for improving scorpion activated protein is lived
Property.
Contain 0.4% 3- hydroxy-5-methyl base -2,4- imidazolones, 3- hydroxyls -5- in ethanol solution in deodorant step
The content containing (R) -3- hydroxy-5-methyl base -2,4- imidazolones is 78% in methyl -2,4- imidazolones；Scorpion activated protein
Aldehyde, amine substance in system have certain oxidisability, may cause certain degradation to scorpion activated protein, and ethyl alcohol
(R) -3- hydroxy-5-methyls base -2,4- imidazolones of specific proportioning and (S) -3- hydroxy-5-methyl base -2,4- imidazolines in solution
There is ketone synergistic effect, the synergistic effect can greatly enhance the dissolubility of aldehyde, amine substance in organic phase in system, and one
Aspect can be rapidly separated out aldehyde, amine substance, improve deodorization effect, on the other hand also reduce aldehyde, amine substance to scorpion
The degradation of activated protein improves the content and purity of scorpion activated protein in final product.
The purposes of scorpion activated protein, purposes of the scorpion activated protein in anti-oxidant nutriment, scorpion activated protein can
Body total antioxidant capacity, activities of antioxidant enzymes and GSH contents are effectively improved, to reduce mistake of the oxygen radical to Cell membrane lipids
Oxidation reduces body MDA contents, shows inhibiting effect to liver spontaneity lipid peroxidation, therefore certainly with removing
By biological effectiveness such as base, antibacterial, anti-oxidant and anti-agings, a kind of environmentally friendly, efficient anti-oxidant green can be applied to
Nutriment.
Embodiment 4：
According to GB5009.5-2016《The measurement of national food safety standard Protein in Food》, measure in scorpion activated protein
Protein content；According to GB5009.4-2016《The measurement of ash content in national food safety standard food》, measure scorpion and live
Content of ashes in property albumen；The crude fat content in remaining method measurement scorpion activated protein is extracted using Soxhlet fat.To reality
The measurement result for applying the scorpion activated protein in a 1-3 is as shown in table 1, as shown in Table 1, compared to embodiment 1, embodiment 2 with
Protein content in embodiment 3 has been more than 90%, and ash content is relatively low with crude fat content, especially protein in embodiment 3
Content has been more than 95%, illustrates that the method for the present invention is a kind of preparation method of efficient scorpion activated protein.
The measurement result of scorpion activated protein in 1. embodiment 1-3 of table
| Project | Embodiment 1 | Embodiment 2 | Embodiment 3 |
| Protein content | 75.8% | 90.2% | 95.8% |
| Content of ashes | 8.3% | 2.5% | 1.1% |
| Crude fat content | 12.5% | 6.6% | 2.8% |
Routine operation in operating procedure of the present invention is well known to those skilled in the art, herein without repeating.
Technical scheme 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 the spirit of the present invention,
Supplement or similar fashion replacement etc., should all be included in the protection scope of the present invention.
Claims (8)
1. the preparation method of scorpion activated protein, including：Degreasing, separation, dialysis, deodorant, it is characterised in that：The separating step
For：Scorpion skimmed milk is weighed in container, the distilled water containing ethyl alcohol, 2-chloroethyl amine hydrochloride and l-tyrosine hydrazides is added,
PH to 11.5~11.8 is adjusted, is centrifuged after magnetic agitation extraction, collection supernatant adjusting pH to 4.5~4.55 precipitating proteins,
It is then centrifuged for collecting sediment, is washed with distilled water sediment, freeze-drying obtains scorpion protein isolate.
2. the preparation method of scorpion activated protein according to claim 1, it is characterised in that：Contain 2 in the distilled water
~3% ethyl alcohol, 0.5~0.6% 2-chloroethyl amine hydrochloride and 0.02~0.03 ‰ l-tyrosine hydrazides.
3. the preparation method of scorpion activated protein according to claim 1, it is characterised in that：The defatting step is：It will
Scorpion meat crushes after cleaning up, and is transferred in vacuum freeze drier after being placed in -80~-90 DEG C of freezings, is faced using super after crushing
Boundary CO2It extracts degreasing and obtains fat-soluble by-product, the degreasing dregs of rice are crushed into chilled storage.
4. the preparation method of scorpion activated protein according to claim 1, it is characterised in that：The dialysis step is：It adopts
It is dialysed to scorpion separated protein powder with 12ku aperture fibers bag filter, dialyse 48~72h, per a dialyzate is changed for 24 hours, receives
Collection precipitation, freezen protective after freeze-drying.
5. the preparation method of scorpion activated protein according to claim 4, it is characterised in that：In the dialysis step, material
Liquor ratio is 1:200~230, dialysis temperature is 1~2 DEG C.
6. the preparation method of scorpion activated protein according to claim 1, it is characterised in that：The deodorant step is：Claim
The scorpion separated protein powder after dialysis is taken, ethanol solution is added, centrifuging and taking precipitates after high speed homogenization, repeats 2~5 postpositions of deodorant
At aeration-drying, sediment spontaneously dries up to scorpion active protein powder.
7. the preparation method of scorpion activated protein according to claim 6, it is characterised in that：Second in the deodorant step
Contain 0.2~0.5% 3- hydroxy-5-methyl base -2,4- imidazolones, 3- hydroxy-5-methyl base -2,4- imidazolones in alcoholic solution
In containing (R) -3- hydroxy-5-methyl base -2,4- imidazolones content be 78~79%.
8. the purposes of scorpion activated protein, it is characterised in that：Purposes of the scorpion activated protein in anti-oxidant nutriment.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117069801A (en) * | 2023-10-17 | 2023-11-17 | 山东省食品药品检验研究院 | Polypeptide with common characteristics of scorpion of spiny thorn and scorpion of strip-shaped plaque and screening method thereof |
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2018
- 2018-04-19 CN CN201810353518.1A patent/CN108740280A/en not_active Withdrawn
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117069801A (en) * | 2023-10-17 | 2023-11-17 | 山东省食品药品检验研究院 | Polypeptide with common characteristics of scorpion of spiny thorn and scorpion of strip-shaped plaque and screening method thereof |
| CN117069801B (en) * | 2023-10-17 | 2023-12-19 | 山东省食品药品检验研究院 | Polypeptide with common characteristics of scorpion of spiny thorn and scorpion of strip-shaped plaque and screening method thereof |
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