CN1274716C - Method for extracting albumin transferrin with iron supplemented and antibacterial function - Google Patents
Method for extracting albumin transferrin with iron supplemented and antibacterial function Download PDFInfo
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- CN1274716C CN1274716C CN 200310112528 CN200310112528A CN1274716C CN 1274716 C CN1274716 C CN 1274716C CN 200310112528 CN200310112528 CN 200310112528 CN 200310112528 A CN200310112528 A CN 200310112528A CN 1274716 C CN1274716 C CN 1274716C
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- egg white
- transferrins
- albumin
- iron complexes
- exchange resin
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Abstract
The present invention discloses a method for extracting albumin transferrin with functions of supplying iron and resisting bacteria. The present invention comprises the following steps: (1) using fowl egg white as raw material, adding ammonium sulfate to make saturation reach 40 to 50%, adjusting pH value to 3.0 to 6.0, statically positioning at 0 to 10 DEG C, carrying out centrifugation or filtration to remove mixed protein deposition mainly containing albumin, adding ammonium sulfate into supernatant liquid to make the saturation reach 60 to 80%, adjusting the pH value to 6.0 to 8.0, statically positioning the supernatant liquid at 0 to 10 DEG C, and collecting albumin deposition; (2) dissolving the albumin deposition obtained in step (1), further removing mixed protein by chromatographic separation to obtain protein eluent; (3) desalting the protein eluent obtained in step (2) by using anion and cation exchange resin, and obtaining albumin transferrin by condensation and freeze-drying. The present invention has the characteristics of simple technology, high product extraction rate and good application prospect.
Description
Technical field
The present invention relates to a kind of extracting method of egg white Transferrins,iron complexes.
Background technology
Transferrins,iron complexes is that cluster has multiple bioactive betaglobulin, mainly comprise three kinds of typical albumen: the egg white Transferrins,iron complexes in birds egg white and the serum (claiming siderophilin again), the Serum transferrin in lactoferrin in the Mammals whey and the vertebrates serum.The tertiary structure in the proteic aminoacid sequence of this bunch, active centre and function are all very similar, the physiological function that all has good enhancing body immunizing power, can promote the absorption of infant to iron and nutritive substance, effectively prevent anaemia, suppress the generation of pathogen enterobacteria and viral interior free yl, alleviate rheumatoid arthritis and anti-ageing.In recent years, abroad developed the lactoferrin in this bunch albumen, be widely used in the middle of the protective foods as important food function factor.
The content of lactoferrin is very little in the cow's milk, and China lacks the whey resource again, and present technology will realize that the suitability for industrialized production difficulty of Transferrins,iron complexes is very big, the extraction cost costliness.Egg white transferrin content in the birds egg white is far above the lactoferrin of bovine whey, and abroad the someone adopts the method for affinity ultrafiltration to separate the Ovum Gallus domesticus album Transferrins,iron complexes, but ultrafiltration process often limits its practical application because of concentration polarization and film pollute.The birds, beasts and eggs dirt of China produces scale and output all ranks first in the world, and utilizes new technology to extract the egg white Transferrins,iron complexes from the birds, beasts and eggs of China's abundant are taken stock of assets the source, and prospect is boundless.
Summary of the invention
The object of the present invention is to provide a kind of extracting method of new egg white Transferrins,iron complexes.
A kind of extracting method provided by the invention with egg white Transferrins,iron complexes of mending the iron anti-microbial effect, this method may further comprise the steps:
(1) is raw material with birds, beasts and eggs, adds ammonium sulfate and make saturation ratio reach 40% that adjust pH is 4.2,4 ℃ and leaves standstill clearly, the foreign protein precipitation based on albumin is removed in centrifugal or filtration, and supernatant liquor adds ammonium sulfate again, makes saturation ratio reach 70%, adjust pH is 6.0,4 ℃ and leaves standstill, and collects albumen precipitation;
(2) albumen precipitation with step (1) gained is dissolved in water, and further removes foreign protein through chromatographic separation, obtains the albumen elutriant;
(3) the albumen elutriant with step (2) gained carries out desalination with anion-cation exchange resin, and concentrated, lyophilize obtains the egg white Transferrins,iron complexes.
In the extracting method of the present invention, the birds, beasts and eggs that step (1) is selected for use are clearly Ovum Gallus domesticus album, Ovum Anas domestica album, goose albumen or quail egg white; It is DEAE-Mierocrystalline cellulose or ion exchange resin that step (2) is separated the used chromatography carrier of foreign protein; The used Zeo-karb of desalination is 724 or 732 Zeo-karbs in the step (3), and anionite-exchange resin is 701 anionite-exchange resin.
The present invention has the advantages that technology is simple, the product extraction yield is high, has a good application prospect.
Embodiment
The extracting method of egg white Transferrins,iron complexes of the present invention may further comprise the steps:
(1) gets freshly-slaughtered poultry egg white 500 gram,, remove by filter wherein insolubles with filter cloth with the dilution of 500 ml waters;
(2) add a small amount of liquor ferri trichloridi, make egg white Transferrins,iron complexes contained in the egg white solution saturated by ferric ion;
(3) add the anhydrous slufuric acid ammonium, the ammonium sulfate saturation ratio that makes solution is 40%, adjusting the pH value of solution value is 4.2,4 ℃ left standstill 12 hours, 2000 leave gains in depth of comprehension to 1000 milliliters of supernatant liquors, continue to add the anhydrous slufuric acid ammonium in the supernatant liquor and make saturation ratio reach 70%, and transferring pH with ammoniacal liquor is 6.0,4 ℃ left standstill 12 hours, and 2000 leave collecting precipitation behind the heart;
(4) with 500 ml water dissolution precipitations, gained solution is to the water desalination of dialysing, again to pH7.0, the dialysis of 0.02mol/LTris-HCl solution;
(5) dialysis back solution is crossed the DEAE-cellulose chromatography, with 0.05mol/L NaCl wash-out, collect elutriant, get egg white Transferrins,iron complexes solution, gained egg white Transferrins,iron complexes liquid is crossed 724 Zeo-karbs earlier, after 701 anionite-exchange resin, remove salt ion, the protein liquid behind the desalination is through lyophilize, obtain egg white Transferrins,iron complexes product 3 grams, product identifies that through the SDS-polyacrylamide gel electrophoresis molecular weight is 76KD, and purity is higher than 80%.
Claims (4)
1, a kind of extracting method with egg white Transferrins,iron complexes of mending the iron anti-microbial effect may further comprise the steps:
(1) is raw material with birds, beasts and eggs, adds ammonium sulfate and make saturation ratio reach 40% that adjust pH is 4.2,4 ℃ and leaves standstill clearly, the foreign protein precipitation based on albumin is removed in centrifugal or filtration, and supernatant liquor adds ammonium sulfate again, makes saturation ratio reach 70%, adjust pH is 6.0,4 ℃ and leaves standstill, and collects albumen precipitation;
(2) albumen precipitation with step (1) gained is dissolved in water, and further removes foreign protein through chromatographic separation, obtains the albumen elutriant;
(3) the albumen elutriant with step (2) gained carries out desalination with anion-cation exchange resin, and concentrated, lyophilize obtains the egg white Transferrins,iron complexes.
2, the extracting method of egg white Transferrins,iron complexes according to claim 1 is characterized in that birds, beasts and eggs used in the step (1) are clearly Ovum Gallus domesticus album, Ovum Anas domestica album, goose albumen or quail egg white.
3, the extracting method of egg white Transferrins,iron complexes according to claim 1 is characterized in that it is DEAE-Mierocrystalline cellulose or ion exchange resin that step (2) is separated the used chromatography carrier of foreign protein.
4, the extracting method of egg white Transferrins,iron complexes according to claim 1 is characterized in that the used Zeo-karb of desalination is 724 or 732 Zeo-karbs in the step (3), and anionite-exchange resin is 701 anionite-exchange resin.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200310112528 CN1274716C (en) | 2003-12-11 | 2003-12-11 | Method for extracting albumin transferrin with iron supplemented and antibacterial function |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200310112528 CN1274716C (en) | 2003-12-11 | 2003-12-11 | Method for extracting albumin transferrin with iron supplemented and antibacterial function |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1546524A CN1546524A (en) | 2004-11-17 |
| CN1274716C true CN1274716C (en) | 2006-09-13 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200310112528 Expired - Fee Related CN1274716C (en) | 2003-12-11 | 2003-12-11 | Method for extracting albumin transferrin with iron supplemented and antibacterial function |
Country Status (1)
| Country | Link |
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| CN (1) | CN1274716C (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2603517A1 (en) * | 2005-04-08 | 2006-11-02 | Snow Brand Milk Products Co., Ltd. | Iron composition containing milk protein |
| CN102604915B (en) * | 2012-03-27 | 2013-08-21 | 华中农业大学 | Method for jointly extracting a variety of proteins from egg white |
| CN105037531A (en) * | 2015-08-10 | 2015-11-11 | 武汉华美生物工程有限公司 | Method for preparing high-purity human plasma transferrin |
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2003
- 2003-12-11 CN CN 200310112528 patent/CN1274716C/en not_active Expired - Fee Related
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| Publication number | Publication date |
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| CN1546524A (en) | 2004-11-17 |
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