CN104479003A - Selenous acidized beta-lactoglobulin, preparation method and application - Google Patents
Selenous acidized beta-lactoglobulin, preparation method and application Download PDFInfo
- Publication number
- CN104479003A CN104479003A CN201410820824.3A CN201410820824A CN104479003A CN 104479003 A CN104479003 A CN 104479003A CN 201410820824 A CN201410820824 A CN 201410820824A CN 104479003 A CN104479003 A CN 104479003A
- Authority
- CN
- China
- Prior art keywords
- lactoglobulin
- beta
- selenous acid
- selenous
- acid beta
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 108010060630 Lactoglobulins Proteins 0.000 title claims abstract description 110
- 238000002360 preparation method Methods 0.000 title claims abstract description 22
- 102000000119 Beta-lactoglobulin Human genes 0.000 title claims 18
- MCAHWIHFGHIESP-UHFFFAOYSA-N selenous acid Chemical group O[Se](O)=O MCAHWIHFGHIESP-UHFFFAOYSA-N 0.000 claims abstract description 93
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 claims abstract description 33
- 229910052711 selenium Inorganic materials 0.000 claims abstract description 33
- 239000011669 selenium Substances 0.000 claims abstract description 33
- 238000004108 freeze drying Methods 0.000 claims abstract description 6
- 229940082569 selenite Drugs 0.000 claims description 20
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 16
- 238000006243 chemical reaction Methods 0.000 claims description 15
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims description 10
- 238000001556 precipitation Methods 0.000 claims description 9
- 239000007800 oxidant agent Substances 0.000 claims description 8
- 238000000108 ultra-filtration Methods 0.000 claims description 8
- 239000012528 membrane Substances 0.000 claims description 7
- 239000004475 Arginine Substances 0.000 claims description 6
- CHJJGSNFBQVOTG-UHFFFAOYSA-N N-methyl-guanidine Natural products CNC(N)=N CHJJGSNFBQVOTG-UHFFFAOYSA-N 0.000 claims description 6
- ZRALSGWEFCBTJO-UHFFFAOYSA-N anhydrous guanidine Natural products NC(N)=N ZRALSGWEFCBTJO-UHFFFAOYSA-N 0.000 claims description 6
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims description 6
- SWSQBOPZIKWTGO-UHFFFAOYSA-N dimethylaminoamidine Natural products CN(C)C(N)=N SWSQBOPZIKWTGO-UHFFFAOYSA-N 0.000 claims description 6
- WQYVRQLZKVEZGA-UHFFFAOYSA-N hypochlorite Chemical compound Cl[O-] WQYVRQLZKVEZGA-UHFFFAOYSA-N 0.000 claims description 6
- 230000008676 import Effects 0.000 claims description 6
- 125000000637 arginyl group Chemical group N[C@@H](CCCNC(N)=N)C(=O)* 0.000 claims description 5
- 238000005406 washing Methods 0.000 claims description 5
- 125000000879 imine group Chemical group 0.000 claims description 4
- XTEGARKTQYYJKE-UHFFFAOYSA-N chloric acid Chemical group OCl(=O)=O XTEGARKTQYYJKE-UHFFFAOYSA-N 0.000 claims description 3
- 229940005991 chloric acid Drugs 0.000 claims description 3
- 239000003814 drug Substances 0.000 claims description 3
- QWPPOHNGKGFGJK-UHFFFAOYSA-N hypochlorous acid Chemical compound ClO QWPPOHNGKGFGJK-UHFFFAOYSA-N 0.000 claims description 3
- 230000036039 immunity Effects 0.000 claims description 3
- 102000008192 Lactoglobulins Human genes 0.000 abstract description 92
- 230000033116 oxidation-reduction process Effects 0.000 abstract description 9
- 230000015572 biosynthetic process Effects 0.000 abstract description 6
- 238000000034 method Methods 0.000 abstract description 6
- 238000011161 development Methods 0.000 abstract description 4
- 230000018109 developmental process Effects 0.000 abstract description 4
- 230000003712 anti-aging effect Effects 0.000 abstract description 3
- 230000000050 nutritive effect Effects 0.000 abstract description 2
- 102000004169 proteins and genes Human genes 0.000 abstract description 2
- 108090000623 proteins and genes Proteins 0.000 abstract description 2
- 239000002994 raw material Substances 0.000 abstract description 2
- 229910018162 SeO2 Inorganic materials 0.000 abstract 1
- 230000001093 anti-cancer Effects 0.000 abstract 1
- 238000005119 centrifugation Methods 0.000 abstract 1
- 230000007166 healthy aging Effects 0.000 abstract 1
- JPJALAQPGMAKDF-UHFFFAOYSA-N selenium dioxide Chemical compound O=[Se]=O JPJALAQPGMAKDF-UHFFFAOYSA-N 0.000 abstract 1
- 238000001308 synthesis method Methods 0.000 abstract 1
- 239000000047 product Substances 0.000 description 16
- 239000000243 solution Substances 0.000 description 15
- 102000007544 Whey Proteins Human genes 0.000 description 8
- 108010046377 Whey Proteins Proteins 0.000 description 8
- 239000007788 liquid Substances 0.000 description 5
- 238000003786 synthesis reaction Methods 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 4
- 239000005862 Whey Substances 0.000 description 4
- 230000006870 function Effects 0.000 description 4
- -1 tin anhydride Chemical class 0.000 description 4
- 235000021119 whey protein Nutrition 0.000 description 4
- 150000002500 ions Chemical class 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 102000008114 Selenoproteins Human genes 0.000 description 2
- 108010074686 Selenoproteins Proteins 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 230000003005 anti-senility effect Effects 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 235000006708 antioxidants Nutrition 0.000 description 2
- 230000006907 apoptotic process Effects 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 235000013351 cheese Nutrition 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 235000013365 dairy product Nutrition 0.000 description 2
- 231100000053 low toxicity Toxicity 0.000 description 2
- 235000013336 milk Nutrition 0.000 description 2
- 239000008267 milk Substances 0.000 description 2
- 210000004080 milk Anatomy 0.000 description 2
- 230000035790 physiological processes and functions Effects 0.000 description 2
- 239000000376 reactant Substances 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- 239000002699 waste material Substances 0.000 description 2
- 102000014914 Carrier Proteins Human genes 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 101000829725 Homo sapiens Phospholipid hydroperoxide glutathione peroxidase Proteins 0.000 description 1
- 208000019926 Keshan disease Diseases 0.000 description 1
- 206010039921 Selenium deficiency Diseases 0.000 description 1
- 238000002441 X-ray diffraction Methods 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 108091008324 binding proteins Proteins 0.000 description 1
- 210000000601 blood cell Anatomy 0.000 description 1
- 230000003925 brain function Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000002983 circular dichroism Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000002354 daily effect Effects 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 238000000113 differential scanning calorimetry Methods 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 235000020776 essential amino acid Nutrition 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000037406 food intake Effects 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 239000002440 industrial waste Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000001665 lethal effect Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000009456 molecular mechanism Effects 0.000 description 1
- 230000000926 neurological effect Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 238000003305 oral gavage Methods 0.000 description 1
- 201000008482 osteoarthritis Diseases 0.000 description 1
- 239000012286 potassium permanganate Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 238000005096 rolling process Methods 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 238000011287 therapeutic dose Methods 0.000 description 1
- 231100000816 toxic dose Toxicity 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
Classifications
-
- 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
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
- C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
- C07K14/4717—Plasma globulins, lactoglobulin
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicinal Chemistry (AREA)
- Zoology (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Gastroenterology & Hepatology (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Toxicology (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention relates to a synthesis and preparation method and an application of selenous acidized beta-lactoglobulin. Beta-lactoglobulin and SeO2 are taken as raw materials, selenous acid groups are introduced into beta-lactoglobulin with an oxidation-reduction method, and a novel organic selenium source is prepared. The preparationmethodcomprises the following steps: preparation of selenous acid; preparation ofa beta-lactoglobulinliquid containing selenous acid; introduction of selenous acid radical; filterationand centrifugation; freeze drying; and obtaining of a pure product selenous acidized beta-lactoglobulin. The obtained selenous acidized beta-lactoglobulin has high nutritive value, stable properties, and anticancer and anti-aging effects, and is an excellent organic selenium source. The method not only has greattheoretical significance on development of organic selenized protein, but also has great social practical significance on healthy aging.
Description
Technical field
The invention belongs to the field of the multi-crossed disciplines such as food scientific technology, organic chemistry and field of biological pharmacy, providing is a kind of selenous acid beta-lactoglobulin and synthesis preparation method and application.
Background technology
Selenium is the requisite trace element of living organism, is antioxidant important in body.Selenium affects the Radical Metabolism of organism, anti-oxidant function, immunologic function, reproductive function and apoptosis etc. by regulating seleno-protein or selenium enzyme (Selenoperoxidase and human phospholipid hydroperoxide glutathione peroxidase).Large quantifier elimination shows both at home and abroad, and the aging of selenium and body is closely related.Selenium, as the attemperator of brain function, participates in multiple with increasing the neurological occurred age.
Selenium in Soil is the main selenium source needed for organism, but the abundance of selenium in the earth's crust is extremely low, its distribution is also extremely unbalanced, and there is more than 40 countries and regions scarce selenium in various degree in the whole world, consequently leads to the generation of many region selenium deficiency diseases, as Keshan disease, Kaschin-Beck disease etc.China is serious one of country lacking selenium in the world, and the region area of about 72% is in scarce selenium state in various degree.Diet cannot meet the physiological requirements of human body, and therefore, the physiological function maintaining human normal depends on the picked-up of external source selenium.But, the therapeutic dose of inorganic selenium and the scope of toxic dose very narrow, and the antioxygenation etc. of selenium often depends on higher dosage.The toxicity ratio inorganic selenium of organoselenium is little, and bioavailability is high, also remarkable than inorganic selenium on challenge.Therefore, the synthesis of organic selenium source of high-efficiency low-toxicity and functional study for maintaining human normal physiological function, delay senility significant.
Beta-lactoglobulin (β-lactoglobulin, β-Lg) is a kind of important whey-protein, accounts for 50% of milk whey protein total amount.Whey had once been considered to the waste of the milk-product such as cheese, cheese processing in dairy industry, was only simply processed into the low value-added product such as whey powder or condensed whey protein.Along with the development of national economy and the raising of people's living standard, dairy industry development rapidly, has become one of important mainstay industry of China's foodstuffs industry.The exploitation of whey by-product enjoy people to pay close attention to.Whey-protein main component beta-lactoglobulin molecular weight is about 18kDa, is made up of 162 amino-acid residues, and containing multiple indispensable amino acid, wherein containing 3 arginine residues, amino acid ratio is reasonable, is a kind of natural quality protein.Because each beta-lactoglobulin molecule contains three arginine residues, can in conjunction with three selenite radicals, if everyone every daily ingestion 10 grams of beta-lactoglobulins, just can take in the selenium of about 0.07 milligram simultaneously, just in time between the normal range 0.05-0.1 milligram of selenium intake, so utilize beta-lactoglobulin to carry out selenizing, the joint development being conducive to beta-lactoglobulin and selenium utilizes.The research of beta-lactoglobulin vacuum regulation and control selenization technology and the anti-ageing molecular mechanism of selenizing beta-lactoglobulin for other organic selenium albumen develop and seleno-protein regulation and control antisenility function Journal of Sex Research has great theory significance, and there is actual application value.
Summary of the invention
The object of the present invention is to provide a kind of selenous acid beta-lactoglobulin and preparation method and application.Selenite radical to be imported on the imine group of beta-lactoglobulin arginine guanidine radicals by oxidation-reduction method and generates selenous acid beta-lactoglobulin by the present invention, and this synthesis preparation method technique is simple, and product yield is high.
The object of the present invention is achieved like this:
A kind of selenous acid beta-lactoglobulin, the imine group of the arginine guanidine radicals of beta-lactoglobulin imports selenite radical, forms selenous acid beta-lactoglobulin.
A preparation method for selenous acid beta-lactoglobulin, step is as follows
(1) prepare selenous acid;
(2) beta-lactoglobulin is joined in selenous acid solution;
(3) the importing of selenite radical: step solution is (2) added oxygenant, in vacuum, 35-45 DEG C, reaction 3-8h, imports the arginine residues on beta-lactoglobulin by selenous acid, generate selenous acid beta-lactoglobulin;
(4) selenous acid beta-lactoglobulin is separated, washs rear acquisition selenous acid beta-lactoglobulin product.
And, when step (3) react terminate in rear solution, have remaining strong oxidizer or selenite radical time, with the ultra-filtration membrane of molecular weight 3000Da-7000Da the strong oxidizer of upper step molecular weight and the selenite radical of failing to import crossed and filter.
And (4) described step obtains selenous acid beta-lactoglobulin with alcohol settling is centrifugal, by precipitation washing with alcohol 2-4 time, remove inorganic selenium completely, after vacuum lyophilization, obtain selenous acid beta-lactoglobulin product.
And described oxygenant is chloric acid, oxymuriate, hypochlorous acid, hypochlorite or hydrogen peroxide.
Selenous acid beta-lactoglobulin is as the application of healthcare products.
Selenous acid beta-lactoglobulin is as the application of makeup.
Selenous acid beta-lactoglobulin improves the application of the medicine of immunity of organisms as preparation.
Advantage of the present invention and positively effect are:
1, raw material beta-lactoglobulin of the present invention derives from industrial waste whey, cost is again reduced while taking full advantage of waste resource, and beta-lactoglobulin is a kind of natural quality milk endogenous binding protein matter be of high nutritive value, the selenous acid beta-lactoglobulin that importing selenite radical obtains wherein can also provide abundant nutrition while supplement selenium, is organic selenium source of high-quality.
2, the technology of preparing of this selenous acid beta-lactoglobulin is the selenous acid utilizing oxidation-reduction phenomenon common in organic sphere to achieve beta-lactoglobulin, in the middle of this preparation method, the oxidation-reduction agent of wide material sources can be used, as chloric acid and oxymuriate, hypochlorous acid and hypochlorite thereof, hydrogen peroxide and potassium permanganate etc. all can make selenous acid be attached on beta-lactoglobulin, the concentration of oxidation-reduction agent can be arbitrary concentration.
3, the preparation technology of selenous acid beta-lactoglobulin of the present invention is relatively simple, and product yield is high; The product property obtained is stablized, and its stability is far away higher than tin anhydride, selenous acid and compound thereof; And can preserve with pulverous form, be easy to store, the healthcare products of the form such as tablet, capsule can be made into.
4, the importing of selenite radical makes the higher structure of beta-lactoglobulin change by the present invention, more group is exposed, adds its solubleness, thus is easier to the absorption promoting body.
5, selenous acid beta-lactoglobulin provided by the invention has remarkable lethal effect for blood cell K562, confirming that under 100-300ug/ml concentration, cultivate 24-72h can reach 50-90% to the inhibiting rate of K562 cell by experiment, there is typical apoptosis feature in K562 cell.
6, selenous acid beta-lactoglobulin provided by the invention has low toxicity high efficiency.This selenous acid beta-lactoglobulin of Mouse oral gavage 50-100mg/kg every day, do not show any abnormalities, its toxicity is starkly lower than SeO
2.
Accompanying drawing explanation
Fig. 1 is selenous acid beta-lactoglobulin structure of the present invention;
Fig. 2 is the selenous acidization reaction of beta-lactoglobulin of the present invention, and Fig. 2-1 is reaction formula, and Fig. 2-2 is amino acid of selenous acid beta-lactoglobulin;
Fig. 3 is the structural formula that beta-lactoglobulin arginine guanidine radicals of the present invention is combined with selenous acid;
Fig. 4 is the infrared colour spectrogram of selenous acid beta-lactoglobulin of the present invention and beta-lactoglobulin;
Fig. 5 is selenous acid beta-lactoglobulin SDS-PAGE gel electrophoresis figure of the present invention;
Fig. 6 is the circular dichroism spectrogram of selenous acid beta-lactoglobulin of the present invention and beta-lactoglobulin;
Fig. 7 is the x-ray diffraction pattern of selenous acid beta-lactoglobulin of the present invention and beta-lactoglobulin;
Fig. 8 is selenous acid beta-lactoglobulin of the present invention (Fig. 8 b) and beta-lactoglobulin (Fig. 8 differential scanning calorimetry figure a);
Fig. 9 is that selenous acid beta-lactoglobulin of the present invention is on the impact of K562 cellular form.(Fig. 9 is 0h a); (Fig. 9 b) 24h; (Fig. 9 c) 48h; (Fig. 9 d) 72h, cultivates concentration and is 200ug/mL.
Embodiment
Describe embodiments of the invention in detail below; It should be noted that, the present embodiment is narrative, is not determinate, can not limit protection scope of the present invention with this.
This patent is intended to prepare organic selenium albumen, then verifies the antisenility function of selenous acid beta-lactoglobulin, for it provides scientific basis in the application in the fields such as medicine, healthcare products, makeup.
Selenous acid beta-lactoglobulin provided by the invention be selenite radical imported to beta-lactoglobulin arginine guanidine radicals by oxidation-reduction method imine group on generate the preparation method of selenous acid beta-lactoglobulin and selenous acid beta-lactoglobulin, this synthesis preparation method technique is simple, product yield is high, and wherein selenous acid beta-lactoglobulin is the selenite radical importing stable in properties on the arginine guanidine radicals of beta-lactoglobulin.
After the present invention also aims to synthesis selenous acid beta-lactoglobulin, further research its in application that is anti-ageing and that improve in immunity of organisms etc.
Selenous acid beta-lactoglobulin of the present invention is completed by following steps:
(1) selenous acid H
2seO
3preparation: tin anhydride SeO
2with water H
2o reaction generates selenous acid H
2seO
3;
(2) the preparation of the beta-lactoglobulin liquid containing selenous acid: beta-lactoglobulin is placed in the excessive selenous acid solution prepared containing above-mentioned steps, obtains the beta-lactoglobulin liquid containing selenous acid;
(3) the importing of selenite radical: under the above-mentioned beta-lactoglobulin liquid containing selenous acid is placed in the ambient condition of the oxidation-reduction containing strong oxidizer, less than vacuum, 35-45 DEG C is reacted 3-8h, selenous acid can be imported the arginine residues on beta-lactoglobulin, generate selenous acid beta-lactoglobulin;
(4) remove the ion in reaction solution after completion of the reaction: with the ultra-filtration membrane of molecular weight 3000Da-7000Da by the strong oxidizer of upper step molecular weight and fail the selenite radical of importing and cross and filter;
(5) the collection of selenous acid beta-lactoglobulin: obtain selenous acid beta-lactoglobulin through the alcohol settling of the selenous acid beta-lactoglobulin different concns of ultrafiltration purification is centrifugal, by precipitation washing with alcohol 2-4 time, remove inorganic selenium completely, after vacuum lyophilization, obtain selenous acid beta-lactoglobulin product.
Embodiment 1
A preparation method for selenous acid beta-lactoglobulin, step is as follows:
(1) getting 5g tin anhydride is dissolved in the water of 50mL, makes selenous acid solution.
(2) get 10g beta-lactoglobulin to mix with above-mentioned selenous acid solution, obtain the beta-lactoglobulin liquid containing selenous acid.
(3) the oxidation-reduction material in above-mentioned reaction solution adopts the hydrogen peroxide of 6%, and above reactant is put into vacuum reactor, and under vacuum, 40 degree are stirred 5h, complete the importing of selenite radical.
(4) remove the ion in reaction solution after completion of the reaction: with the ultra-filtration membrane of molecular weight 3000Da-7000Da by the strong oxidizer of upper step molecular weight and fail the selenite radical of importing and cross and filter;
(5) in reaction solution, add 100mL ethanol, centrifugal, obtain product precipitation, successively by 20mL washing with alcohol precipitation, in triplicate, obtain selenous acid beta-lactoglobulin after precipitation vacuum lyophilization, product selenium content is 15.17mg/g.
(6) through detecting: the character of selenous acid beta-lactoglobulin is:
1, the aqueous solution of selenous acid beta-lactoglobulin is transparent, colourless, or yellowish;
2, the solvability of selenous acid beta-lactoglobulin: alkali is molten, salt is molten, is slightly soluble in water, is insoluble to the organic reagent such as ethanol, acetone.
3, selenous acid beta-lactoglobulin is after lyophilize, in white powder;
4, molecular weight: 20.75KDa;
5, selenium content: 15.17mg/g.
Embodiment 2
A preparation method for selenous acid beta-lactoglobulin, step is as follows:
(1) getting 5g tin anhydride is dissolved in the water of 50mL, makes selenous acid solution.
(2) get 20g beta-lactoglobulin to mix with above-mentioned selenous acid solution, obtain the beta-lactoglobulin liquid containing selenous acid.
(3) the oxidation-reduction material in above-mentioned reaction solution adopts the hydrogen peroxide of 8%, and above reactant is put into vacuum reactor, and under vacuum, 45 degree are stirred 7h, complete the importing of selenite radical.
(4) remove the ion in reaction solution after completion of the reaction: with the ultra-filtration membrane of molecular weight 3000Da-7000Da by the strong oxidizer of upper step molecular weight and fail the selenite radical of importing and cross and filter;
(5) in reaction solution, add 200mL ethanol, centrifugal, obtain product precipitation, successively by 40mL washing with alcohol precipitation, repeat four times, obtain selenous acid beta-lactoglobulin after precipitation vacuum lyophilization, product selenium content is 12.53mg/g.
(6) through detecting: the character of selenous acid beta-lactoglobulin is:
1, the aqueous solution of selenous acid beta-lactoglobulin is transparent, colourless, or yellowish;
2, the solvability of selenous acid beta-lactoglobulin: alkali is molten, salt is molten, is slightly soluble in water, is insoluble to the organic reagent such as ethanol, acetone.
3, selenous acid beta-lactoglobulin is after lyophilize, in white powder;
4, molecular weight: 41.06KDa;
5, selenium content: 12.53mg/g.
To be 1812 type rolling ultrafiltration membranes produced by the Shanghai fast science equipment company limited that rubs for the proposed model that the ultra-filtration membrane that above-described embodiment step (4) uses is and producer.
Claims (8)
1. a selenous acid beta-lactoglobulin, is characterized in that: on the imine group of the arginine guanidine radicals of beta-lactoglobulin, import selenite radical, forms selenous acid beta-lactoglobulin.
2. a preparation method for selenous acid beta-lactoglobulin, is characterized in that: step is as follows
(1) prepare selenous acid;
(2) beta-lactoglobulin is joined in selenous acid solution;
(3) the importing of selenite radical: step solution is (2) added oxygenant, in vacuum, 35-45 DEG C, reaction 3-8h, imports the arginine residues on beta-lactoglobulin by selenous acid, generate selenous acid beta-lactoglobulin;
(4) selenous acid beta-lactoglobulin is separated, washs rear acquisition selenous acid beta-lactoglobulin product.
3. the preparation method of selenous acid beta-lactoglobulin according to claim 2, it is characterized in that: when step (3) react terminate in rear solution, have remaining strong oxidizer or selenite radical time, with the ultra-filtration membrane of molecular weight 3000Da-7000Da the strong oxidizer of upper step molecular weight and the selenite radical of failing to import crossed and filter.
4. the preparation method of selenous acid beta-lactoglobulin according to claim 2, it is characterized in that: (4) described step obtains selenous acid beta-lactoglobulin with alcohol settling is centrifugal, by precipitation washing with alcohol 2-4 time, remove inorganic selenium completely, after vacuum lyophilization, obtain selenous acid beta-lactoglobulin product.
5. the preparation method of selenous acid beta-lactoglobulin according to claim 2, is characterized in that: described oxygenant is chloric acid, oxymuriate, hypochlorous acid, hypochlorite or hydrogen peroxide.
6. a selenous acid beta-lactoglobulin according to claim 1 is as the application of healthcare products.
7. a selenous acid beta-lactoglobulin according to claim 1 is as the application of makeup.
8. a selenous acid beta-lactoglobulin according to claim 1 is as the application preparing the medicine improving immunity of organisms.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410820824.3A CN104479003A (en) | 2014-12-25 | 2014-12-25 | Selenous acidized beta-lactoglobulin, preparation method and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410820824.3A CN104479003A (en) | 2014-12-25 | 2014-12-25 | Selenous acidized beta-lactoglobulin, preparation method and application |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104479003A true CN104479003A (en) | 2015-04-01 |
Family
ID=52753624
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410820824.3A Pending CN104479003A (en) | 2014-12-25 | 2014-12-25 | Selenous acidized beta-lactoglobulin, preparation method and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104479003A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105732794A (en) * | 2015-11-06 | 2016-07-06 | 广西大学 | Method of recyclable large-scale separation of [beta]-lactoglobulin |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101715869A (en) * | 2009-11-25 | 2010-06-02 | 云南大学 | Novel method for preparing selenate food protein |
| CN101768097A (en) * | 2009-09-21 | 2010-07-07 | 天津科技大学 | Method for preparing selenic acid arginine and application of selenic acid arginine |
| CN102219720A (en) * | 2011-04-28 | 2011-10-19 | 天津市食品加工工程中心 | Novel selenic acid biological material with anti-aging function |
-
2014
- 2014-12-25 CN CN201410820824.3A patent/CN104479003A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101768097A (en) * | 2009-09-21 | 2010-07-07 | 天津科技大学 | Method for preparing selenic acid arginine and application of selenic acid arginine |
| CN101715869A (en) * | 2009-11-25 | 2010-06-02 | 云南大学 | Novel method for preparing selenate food protein |
| CN102219720A (en) * | 2011-04-28 | 2011-10-19 | 天津市食品加工工程中心 | Novel selenic acid biological material with anti-aging function |
Non-Patent Citations (2)
| Title |
|---|
| GUO-BIN DING ET AL: "Preparation and biological evaluation of a novel selenium-containing exopolysaccharide from Rhizobium sp. N613", 《CARBOHYDRATE POLYMERS》 * |
| GUOQIANG ZHENG ET AL: "Selenium modification of b-lactoglobulin (b-Lg) and its biological activity", 《FOOD CHEMISTRY》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105732794A (en) * | 2015-11-06 | 2016-07-06 | 广西大学 | Method of recyclable large-scale separation of [beta]-lactoglobulin |
| CN105732794B (en) * | 2015-11-06 | 2020-07-07 | 广西大学 | Method for circularly separating β -lactoglobulin in large scale |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Dong et al. | Composition and distribution of the main active components in selenium-enriched fruit bodies of Cordyceps militaris link | |
| CN102172393A (en) | Calcium supplement of fish scale protein peptide calcium chelate and preparation method thereof | |
| CN105012940B (en) | A kind of preparation method of nanometre collagen peptide chelated zinc | |
| CN102174073A (en) | Oyster protein peptide and zinc chelate and method for preparing same | |
| CN106191183B (en) | The preparation method and applications of broccoli protein peptides | |
| CN102532339A (en) | Method for selenizing pilose asiabell root polysaccharides and application of product | |
| CN102934810A (en) | Application of chlorella in foods | |
| CN101575359A (en) | Extraction method of water-soluble spirulina phycobiliprotein | |
| CN105029632A (en) | Suaeda salsa fermented beverage and preparation method thereof | |
| CN104757560A (en) | Glutathione yeast enzymolysis solution composition and preparation method thereof | |
| CN110172491B (en) | Multi-element biological protein peptide and preparation method thereof | |
| CN101979655B (en) | Enzyme method for producing mung bean peptide | |
| CN104711203A (en) | Selenium-rich germanium yeast powder | |
| CN103271166A (en) | Soybean whey composite powder and preparation method thereof | |
| CN105029347A (en) | High-nutrient plant salt with various flavoring functions and preparation method thereof | |
| CN107156747A (en) | A kind of cordyceps sinensis walnut peptide nutrient powder and preparation method thereof | |
| CN114134190A (en) | Preparation method of zein active peptide-carried calcium ion nano chelate | |
| CN103865816A (en) | Yeast powder rich in selenium and germanium | |
| CN104479003A (en) | Selenous acidized beta-lactoglobulin, preparation method and application | |
| CN111393408A (en) | Method for preparing natural flavone selenium | |
| CN102987408B (en) | Method for extracting nutritional ingredients such as free amino acids from waste cordyceps militaris cocoons | |
| CN107348278A (en) | A kind of corn soda water and preparation method thereof | |
| CN107699602A (en) | It is a kind of that industrial process and application rich in machine ferrous iron donkey-hide gelatin peptide are prepared by donkey-hide gelatin | |
| CN106858614A (en) | Compound amino acid selenium yeast oral liquid | |
| CN105767997A (en) | Bee pollen and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| EXSB | Decision made by sipo to initiate substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20150401 |