CN1847404A - Technological process of extracting lactoferritin Lf from human foremilk and its product - Google Patents
Technological process of extracting lactoferritin Lf from human foremilk and its product Download PDFInfo
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- CN1847404A CN1847404A CN 200510024866 CN200510024866A CN1847404A CN 1847404 A CN1847404 A CN 1847404A CN 200510024866 CN200510024866 CN 200510024866 CN 200510024866 A CN200510024866 A CN 200510024866A CN 1847404 A CN1847404 A CN 1847404A
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Abstract
The present invention provides extracting process and product of high purity lactoferritin (Lf) from human foremilk. The extracting process includes the following steps: collecting human foremilk of health parturient, centrifuging to eliminate impurity and fatty, eliminating casein from curd, collecting the supernatant as the lactalbumin, processing with 300PS membrane, vacuum concentrating, and freeze drying to obtain extracted Lf with purity of 98 %. The high purity Lf product includes Lf and additive, which may be immunoglobulin, amino acid, Chinese herbal medicine, biological preparation, milk product or their composition, and may be in tablet, capsule or oral liquid.
Description
Technical field
The invention belongs to the utilization biotechnology from the first Ruzhong of human mother's milk, the technical field of separation and Extraction lactoferrin Lf.Be particularly related to a kind of extraction process and goods thereof of extractive of human colostrum lactoferrin Lf.
Background technology
1, the basic structure of lactoferrin
Lactoferrin is that Mammals (people, ox, horse, sheep) colostrum forms stage, lactation period, one of main glycoprotein in the breast phase breast glandular secretion thing that dries up.It is a kind of protein with various biological function, it not only participates in the transhipment of iron, but also have antimicrobial, anti-oxidant, antitumor, strong pain-relieving and adjusting functions of immune system, be considered to a kind of novel antibacterial sterilization, the food and feed additive of anticancer unsting medicine and great exploitation potential for its.
Lactoferrin takes on a red color because of its crystal and claims " red eggs are white ", be a kind of iron associativity glycoprotein, relative molecular mass 77000 ± 2000, its main body is stockless Ginkgo Leaf shape structure arranged side by side, and the size that iron ion is combined in incision part (the Fe interionic is every 2.8～4.3 μ m) oval leaf of two leaves is 5.5 μ m * 3.5 μ m * 3.5 μ m.Contain 2 iron combining sites in the 1 molecule lactoferrin, 15～16 seminoses, 5～6 semi-lactosis, 10～11 acetylglucosamines and 1 sialic acid, wherein neutral sugar content 8.5%, aminosugar content 2.7%.Human milk Lf polypeptide chain is made up of 703 amino-acid residues, has two carbohydrate side chains, carbohydrate content about 7% on it.And L-glutamic acid among the human milk Lf (Glu), asparagus fern door propylhomoserin (Asp), leucine (Lue) and L-Ala (Ala) content are higher, and contained unique sulfur-containing amino acid is halfcystine (Cys), and terminal has a L-Ala (Ala) residue.The human lactoferrin per molecule has two glycosyls and two kinds of molecular conformations, and its main difference is the carbohydrate difference that they are contained.2 melts combine sites are arranged in the molecule, and each site can be in conjunction with a Fe
3+With a HCO
3 -Or CO
3 -2According to lactoferrin in conjunction with Fe
3+Number, it can be divided into iron deficiency type lactoferrin, iron semi-saturation type lactoferrin and three kinds of forms of iron saturation type lactoferrin.Multi-form lactoferrin has different antitypy abilities.
2, the function of lactoferrin and the mechanism of action
The combined function of A, lactoferrin
Lactoferrin can be bases of its various biological function with many material bonded attributes.Contain two Fe in the saturated lactoferrin of per molecule
3+Ion.Lactoferrin can not only be in conjunction with Fe
3+, and to many transition metal such as Cu
2+, Ca
2+, Al
3+Keying action is all arranged.Lactoferrin has been avoided OH in the human body to the combination of iron
+The generation of objectionable impurities.Lactoferrin can participate in cytophagous killing effect by the formation of catalysis free radical.Studies show that iron ion is the necessary material of nearly all bacterial growth except that milk-acid bacteria, and lactoferrin exactly can be captured the required Fe of bacterial growth by rolling into a ball competition iron with the close iron-based of need iron bacteria
3+Thereby, the growth of bacterium is suppressed.
In addition, lactoferrin can also combine with numerous protein and DNA, as casein, albumin, immunoglobulin A and N,O-Diacetylmuramidase and beta-lactoglobulin.Lactoferrin can also combine with cell, all contains the human lactoferrin acceptor in many cells such as intestinal cells, lymphocyte, thrombocyte and breast epithelial cancer cells and the many microorganism cells adventitias.
B, lactoferrin antibiotic presses down malicious function
Liver cell virus (HCV) is a kind of single strand RNA virus that cyst membrane is arranged, and it can cause chronic hepatitis, liver cirrhosis and liver cancer.Lactoferrin can combine with target cell by stoping virus on the envelope protein that is combined in hepatovirus C; The genome of rotavirus is 10 sections different RNA fragments that are wrapped in 3 layers of capsid, protects from infection thereby lactoferrin can stop virus to be adsorbed on the target cell.Lactoferrin also has stronger restraining effect to HIV (human immunodeficiency virus) (HIV).Nearly all research shows that all it is not to stop virus replication after infection that lactoferrin stops virus infected cell, stops its target cell infection but directly be incorporated on all kinds of virus particle.Another sterilization mode of lactoferrin is to realize by a kind of polypeptide that is called lactoferrin antimicrobial peptide.Lactoferrin is through the product of acid egg from enzyme liberating, and its anti-microbial activity exceeds 20 times than lactoferrin, separablely from this degradation product goes out the polypeptide higher 400 times than the anti-microbial activity of lactoferrin, and this peptide species is called as newborn iron polypeptide element (Lfcin).In addition, lactoferrin can also destroy the adventitia of G-bacterium, changes the permeability of after birth and causes the release of lipopolysaccharides (LPS), and play germ-resistant effect.
The immunoloregulation function of C, lactoferrin
Lactoferrin can promote the sterilization and the phagolysis of neutrophilic leukocyte or scavenger cell, and the breeding of Nk cell activity and lymphocyte, neutrophilic leukocyte is had regulating effect.
D, lactoferrin anti-oxidant function
The Fe of some biological reductants such as xitix
2+Or Fe
3+Salt is the lipid oxidation inductor, and lactoferrin can be controlled the catalytic free radical of iron and form reaction by removing the environment iron ion, suppresses the lipid oxidation that iron ion participates in.
Other function of E, lactoferrin
In addition, along with the develop rapidly of biotechnology, the recent findings lactoferrin also has the function of antitumor cell.It also has superpower pain relieving and analgesia function.
Therefore, lactoferrin is the important active substances that is used for development functionality food.Have high medical and nutritive value, will wide application prospect be arranged in fields such as biologic medical, nutritional supplementation, food interpolations.
3, the preparation of lactoferrin and goods thereof
As everyone knows, lactoferrin is present in nearly all mammiferous milk, and just milk content is the highest with the people especially, and the lactoferrin content in the human milk is about ten times of bovine coloctrum, accounts for 20% of total protein.Has very high purification utility value.And the content of other Mammals milk is just lower, does not possess the possibility that industrialization is purified.Although Mammals lactoferrin amino acid has suitable homology, still there is certain difference in the lactoferrin of different sources on aminoacid sequence, and very big difference is also arranged on the effect of biological utilisation.
At present, the technology of extraction separation homology, high purity lactoferrin from human colostrum, Shang Weijian has report.
Summary of the invention
The invention provides a kind of extractive of human colostrum-lactoferrin (Lf) extraction process, its final purpose is in order to obtain human colostrum lactoferrin (Lf) extract.
A further object of the present invention is to provide a kind of lactoferrin (Lf) goods.
In order to achieve the above object, the present invention adopts following technical scheme:
1, collects unnecessary colostrum in the healthy puerpera 1～15 day, in collection process, can adopt inspection methods such as pH value, proportion, sense organ with the control raw materials quality.
2, centrifugal, in order to reach the purpose of removing impurity and fat better, described centrifugation step can be divided into for two steps, at first under 8～12 ℃, and with the speed of 600～800r/min centrifugal about 10 minutes, to remove impurity; Then under 30～40 ℃, with the speed of 2500～3000r/min centrifugal about 10 minutes, to remove impurity and fat;
3, pasteurization, under 63.5 ℃, 30min is with killing microorganisms and virus for process;
4, curdled milk can adopt under 36 ℃ and send out curdled milk with enzyme, also can adopt microbial method or ion exchange method to remove casein;
5, with the grumeleuse fragmentation after the curdled milk, can adopt under 4 ℃, collect whey with the method that the speed of 7000r/min is centrifugal about 20 minutes;
6, the ultra filtration film is handled, and adopts aperture 300PS, 1000PS film to handle to collect whey and sees through liquid;
7, vacuum concentration;
8, lyophilize obtains lactoferrin (Lf) lyophilisate.
Can obtain extractive of human colostrum lactoferrin (Lf) by the inventive method.
Described a kind of lactoferrin (Lf) goods.Comprise the lactoferrin Lf extract and the additive that obtain by aforesaid method, its mass percent is respectively lactoferrin Lf extract 15～80%, additive 15～80%, wherein said additive can be one of immunoglobulin (Ig), amino acid, herbal medicine, biotechnological formulation, all kinds of dairy products or its combination.In the described additive, the mass percent of each component can be all kinds of dairy products 0～5%, immunoglobulin (Ig) 5～80%, amino acid 0～15%, herbal medicine 0～15%, biotechnological formulation 0～15%.
Described immunoglobulin (Ig) can be IgA, IgG.Can be serotype, also can be secretor type.
Described amino acid can be one of taurine, 18 kinds of essential amino acids or its combination.
Described herbal medicine can be genseng, the Radix Astragali, aloe, wolfberry fruit.Can be that former medicine also can be an extract.
Described biotechnological formulation can be all kinds of somatomedins, two qi or lactobacillus, microbiotic, antiviral.
Described all kinds of dairy products can be bovine coloctrum pulvis, animal/vegetable protein pulvis.
The additive formula of the finished product of the present invention is scientific and reasonable, and is with strong points.As Immunoglobulin IgA, IgG, show after deliberation with act synergistically antibacterial virus killing effect of lactoferrin and use apparently higher than folk prescription; Amino acid combines utilization to improving body immunity with lactoferrin, gain in strength, and improves nerve conduction and sight function plays an important role; The former medicine of herbal medicine genseng, the Radix Astragali, aloe, wolfberry fruit, particularly activeconstituents that therefrom extracts and lactoferrin prescription utilization, can significantly improve the activity of human natural killer cell (NK) cell and complement, strengthen neutrophilic leukocyte or microphage phagocytic activity and killing action to virus, thereby improve the immunological competence of body, the generation and the development of prevention and inhibition tumour; Research shows that also lactoferrin (Lf) can to reduce drug dose, reduce microbiotic and antiviral (fungi preparation) infringement to people liver, renal function with all kinds of microbiotic and antiviral synergy.Recently scientist finds that also lactoferrin (Lf) has extremely strong analgesia effect, and the morphine that outclass present widespread use is more than thousand times, and it is to be difficult for habituation, does not have toxic side effect.Can be developed to nutrition (health care) food fully, use late on one's body the cancer patients, help them to overcome insufferable cancerous swelling pain in last date at life.Obviously the additive formula of the finished product of the present invention is scientific and reasonable, with strong points.
Embodiment
Below in conjunction with three human colostrum lactoferrin Lf extraction process embodiment the present invention is described.Among these three extraction process embodiment, except that the parameter that table one is listed, all the other are all identical with following processing step.
Human colostrum lactoferrin Lf extraction process may further comprise the steps:
1, gets the unnecessary colostrum of healthy puerpera, adopt inspection method control raw materials quality such as pH value, proportion, sense organ;
2, centrifugal removal cell and fat;
3, centrifugal removal fat, and in refrigeration below-25 ℃;
4, pasteurization, under 63.5 ℃, through 30min, killing microorganisms and virus;
5, the refrigerated skimming milk is at room temperature thawed after, curdled milk is to remove casein;
6, with the grumeleuse fragmentation under 4 ℃, went precipitation in centrifugal 20 minutes with 7000r/min, collect whey liquid.
7, ultra filtration film is handled whey liquid with the 300PS film, be that original volume two/for the moment, adding repeats this operation four times with capacity physiological saline in trapped fluid, collects trapped fluid when seeing through liquid.
What 8, aforesaid method is obtained holds back (concentrating) liquid, handles through the film of 1000PS again, collects through liquid.
9, vacuum concentration;
10, obtain the lactoferrin lyophilized powder after the lyophilize.
Table one
| Step | Example one | Example two | Example three |
| 2 is centrifugal | 8 ℃ of 600r/min 10 minutes | 10 ℃ of 700r/min 10 minutes | 12 ℃ of 800r/min 10 minutes |
| 3 is centrifugal | 30 ℃ of 2500r/min 10 minutes | 35 ℃ of 2800r/min 10 minutes | 40 ℃ of 3000r/min 10 minutes |
| 5 curdled milks | 36 ℃ of curdled milk enzyme process | Microbial method | The ion-exchanger method |
| 7 ultrafiltration | The 300PS film | The 300PS film | The 300PS film |
| 8 ultrafiltration | The 1000PS film | The 1000PS film | The 1000PS film |
All can obtain extractive of human colostrum lactoferrin Lf pulvis by above-mentioned three embodiment.
Extractive of human colostrum lactoferrin Lf goods embodiment.
Because above-mentioned three extraction process embodiment all can be used as the major ingredient of goods,,, make goods by adding additive as the major ingredient of goods embodiment at this extractive of human colostrum Lf that only obtains with embodiment one.Wherein said additive can be immunoglobulin (Ig), amino acid, herbal medicine, biotechnological formulation, all kinds of dairy products.Below tabulate extractive of human colostrum lactoferrin Lf goods for nine goods embodiment.
Table two
| Embodiment | Major ingredient lactoferrin mass percent | The immunoglobulin (Ig) mass percent (annotating 1) | Amino acid masses per-cent (annotating 2) | The herbal medicine mass percent (annotating 3) | The biotechnological formulation mass percent (annotating 4) | All kinds of dairy quality per-cents (annotating 5) |
| Example one | 75％ | 5％ | 5％ | 5％ | 5％ | 5％ |
| Example two | 75％ | 5％ | 15％ | 0％ | 0％ | 5％ |
| Example three | 75％ | 5％ | 0％ | 15％ | 0％ | 5％ |
| Example four | 75％ | 5％ | 0％ | 0％ | 15％ | 5％ |
| Example five | 15％ | 80％ | 0％ | 0％ | 0％ | 5％ |
| Example six | 80％ | 5％ | 5％ | 5％ | 0％ | 5％ |
| Example seven | 80％ | 5％ | 0％ | 5％ | 5％ | 5％ |
| Example eight | 80％ | 5％ | 5％ | 5％ | 0％ | 5％ |
| Example nine | 50％ | 35％ | 3％ | 4％ | 3％ | 5％ |
The finished product additive of above-mentioned nine embodiment is scientific and reasonable, with strong points.
[annotating one] described immunoglobulin (Ig) can be IgA, IgG.Can be serotype, also can be secretor type.
[annotating two] described amino acid can be one of taurine, 18 kinds of essential amino acids or its combination.
[annotating three] herbal medicine can be genseng, the Radix Astragali, aloe, wolfberry fruit.Can be that former medicine also can be an extract.
[annotating four] biotechnological formulation can be all kinds of somatomedins, two qi or lactobacillus, microbiotic, antiviral.
[annotating five] described all kinds of dairy products can be bovine coloctrum pulvis, animal/vegetable protein pulvis.
Claims (10)
1, a kind of human colostrum extracts the extraction process of no lactoferrin Lf, comprises following processing step:
Collect the unnecessary colostrum of healthy puerpera;
Centrifugal decon, fat;
Pasteurization;
Curdled milk is removed casein;
Supernatant liquor after the collection curdled milk;
The ultra filtration film is handled;
Vacuum concentration;
Lyophilize.
2, technological process of extracting lactoferritin Lf from human foremilk according to claim 1 is characterized in that:
Describedly centrifugally remove impurity, fat was divided into for two steps, at first under 8～12 ℃, with 600～800r/min centrifugal 10 minutes, removed cell and impurity; And then under 30～40 ℃, removed fat in centrifugal 10 minutes with 2500～3000r/min.
3, technological process of extracting lactoferritin Lf from human foremilk according to claim 1 and 2 is characterized in that: the temperature of described pasteurization is 63.5 ℃, and the time is 30 minutes.
4, technological process of extracting lactoferritin Lf from human foremilk according to claim 1 and 2 is characterized in that:
In the described curdled milk step, the curdled milk method adopts one of 36 ℃ of following curdled milk enzyme process, microbial method or ion exchange methods method.
5, technological process of extracting lactoferritin Lf from human foremilk according to claim 1 and 2 is characterized in that:
Described ultra filtration film is handled, and is successively to adopt 300PS and 1000PS membrane processing method.
6, technological process of extracting lactoferritin Lf from human foremilk according to claim 1 and 2 is characterized in that:
Described concentration method is to adopt the method for vacuum concentration to handle.
7, technological process of extracting lactoferritin Lf from human foremilk according to claim 1 and 2 is characterized in that:
Described freeze drying process is the frozen drying method under adopting-40 ℃.
8, a kind of lactoferrin formulations comprises lactoferrin Lf and additive, it is characterized in that:
The described method of arbitrary claim is made in the described lactoferrin Lf extract employing claim 1 to 7, described additive can be one of immunoglobulin (Ig), amino acid, herbal medicine, biotechnological formulation, all kinds of milk-product or its combination, its mass percent is respectively lactoferrin extract 15～80%, additive 15～80%.
9, lactoferrin formulations according to claim 8 is characterized in that: described immunoglobulin (Ig) can be IgA, IgG, can be serotype, also can be secretor type; Described amino acid can be taurine or 18 kinds of essential amino acids; Described herbal medicine can be genseng, the Radix Astragali, aloe, wolfberry fruit.Can be that former medicine also can be an extract; Described biotechnological formulation can be all kinds of somatomedins, two qi or lactobacillus, microbiotic, antiviral; Described all kinds of dairy products can be bovine coloctrum pulvis, animal/vegetable protein pulvis.
10, lactoferrin formulations according to claim 8 is characterized in that: in the described additive, the mass percent of each component is respectively immunoglobulin (Ig) 5～80%, amino acid 0～15%, herbal medicine 0～15%, biotechnological formulation 0～15%, all kinds of dairy products 0～5%.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101117351B (en) * | 2007-04-30 | 2010-08-18 | 北京济普霖生物技术有限公司 | Method for purifying restructuring lactoferrin from transgenic cow's milk |
| CN101731631B (en) * | 2010-01-28 | 2012-11-07 | 王焕杰 | Health-care product composition |
| CN103025347A (en) * | 2010-04-23 | 2013-04-03 | 普若拜特有限公司 | Pharmaceutical compositions |
-
2005
- 2005-04-04 CN CN 200510024866 patent/CN1847404A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101117351B (en) * | 2007-04-30 | 2010-08-18 | 北京济普霖生物技术有限公司 | Method for purifying restructuring lactoferrin from transgenic cow's milk |
| CN101731631B (en) * | 2010-01-28 | 2012-11-07 | 王焕杰 | Health-care product composition |
| CN103025347A (en) * | 2010-04-23 | 2013-04-03 | 普若拜特有限公司 | Pharmaceutical compositions |
| CN105727286A (en) * | 2010-04-23 | 2016-07-06 | 普若拜特有限公司 | Pharmaceutical composition |
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