CN101712974A - Method for preparing egg albumin polypeptide - Google Patents
Method for preparing egg albumin polypeptide Download PDFInfo
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- CN101712974A CN101712974A CN200810121543A CN200810121543A CN101712974A CN 101712974 A CN101712974 A CN 101712974A CN 200810121543 A CN200810121543 A CN 200810121543A CN 200810121543 A CN200810121543 A CN 200810121543A CN 101712974 A CN101712974 A CN 101712974A
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Abstract
The invention discloses a method for preparing egg albumin polypeptide, which comprises the following steps: taking egg white, adding an organic solvent and a precipitant into the egg white with stirring, then standing and centrifugalizing the mixture, adding cold acetone in a volume which is two times that of the supernatant into the supernatant, and fully stirring and centrifugalizing the mixture; combining precipitates obtained twice to form a crude egg albumin product; adding distilled water into the precipitate, adjusting the pH to between 4.5 and 5.0, soaking the precipitate in the distilled water for 1 to 2 hours, filtering the mixture, adding the distilled water into the mixture to ensure that the concentration of albumin is 5 percent, adding complex enzyme into the mixture, intermittently heating and treating the mixture for 5 minutes with microwave, and then inactivating the enzyme; and filtering, condensing and vacuum-drying the enzyme-inactivated enzymolysis solution so as to obtain a product, namely an egg albumin polypeptide product.
Description
Technical field
The invention relates to a kind of preparation method of egg albumin polypeptide, belong to biological technical field.
Background technology
Albumin claims white protein again, is the maximum protein of content in the blood plasma, accounts for 55% of total protein, and relative molecular mass is less.Albumin polypeptide is a kind of function macro-molecular protein in the human plasma, is to keep plasma osmolarity, increase Q volume of blood, provides balance amino acid whose important component, is the indispensable nutritive substance of human body.
Ovalbumin is a protein main in the egg, accounts for 85% of dry weight.The composition of amino acid of pure albumen of ovum gallinaceum and human serum albumin is closely similar, has 20 whole seed amino acids, is present classic protein, can replace albumin to carry out a series of physiological activity.Its primary structure is made up of 385 amino acid; molecular weight is 42699Da; contain four halfcystines and the spherical sugar-protein that the Gelucystine disulfide linkage constitutes; the sequence of C end is Lys-Val-Ser-Pro-COOH; the glycine (Gly) of N end is an acetylation modification, and iso-electric point is 4.5, and the analysis of PAGE electrophoretogram has 3 bands; because contain the balance amino acid of needed by human, its biological value is up to 94%.On the other hand, contain some special peptide matter in the polypeptide chain of ovalbumin, select suitable proteolytic ferment can obtain the biologically active peptides that some have special physiological regulation function.Once more, egg is normal edible a kind of food during people live, its wide material sources, good and cheap.If can carry out deep processing to it, then promote the practical value of egg greatly.
Confirm after deliberation, as the functional activity peptide, albumin polypeptide has good biology performance, enormous function will be brought into play in health care aspect to human body, its biological activity mainly shows following several respects: 1, easily digestion and absorption, promote the functional rehabilitation of immuning tissue, recover thymus gland, spleen, functions such as lymph, strengthening immunity 2, adjust intestinal microflora, improve efficiency of food conversion 3, have the effect that promotes that nucleosides absorbs and turns round, and be increased in the distribution of liver and thymic tissue, promote calcium, iron, the absorption 4 of materials such as nucleic acid, improve hypoproteinemia, promote operative rehabilitation after being ill, small peptide nutrient is provided, preserves moisture, skin care and beauty treatment.5. when it produces biological effect in vivo, can't cause allergic reaction, recover after being ill and the functions of intestines and stomach impaired patients has significant using value for infant, the elderly, pregnant woman.
Now, also more and more to albuminous research, people constantly seek practical more economically method and develop the albumin series product.Confirm after deliberation, the further hydrolysis of albumin is obtained polypeptide products, more help absorbing, and aspect people's health care, will bring into play bigger effect.
The hydrolysis albumin is used chemical degradation method and enzyme liberating method always at present.The chemical degradation method condition is violent, and the proteinaceous nutrient loss is big, thereby now less employing.And the enzyme liberating method can produce specific peptide by the location hydrolysis under mild conditions, and safe and reliable, has no side effect, and therefore albuminous directional enzymatic technology has become the focus of domestic and international research.Present existing research majority is to adopt single enzyme to be used for protein hydrolysate to obtain polypeptide or amino acid product.Single enzyme usually the incomplete characteristics of existence effect and action time longer.Because the protein volatility requires harshness to temperature of reaction, single enzymolysis needs more than the 6h mostly.Character to product has caused bad influence like this.We adopt microwave heating in conjunction with the combinative enzyme hydrolysis method, not only increase the protein catalytic site, overcome the incomplete shortcoming of single enzyme effect, and the reaction conditions gentleness, shorten the reaction times greatly, can fast and effectively albumin have been resolved into the low polypeptide of molecular weight.
Summary of the invention
One of purpose of the present invention provides a kind of preparation method of egg albumin polypeptide.
Two of purpose of the present invention provides the physiological regulation function of albumin polypeptide and biologic activity explanation.
The said albumin polypeptide of the present invention be a kind of be raw material with the fresh hen egg, be divided into two steps: at first be the fresh hen egg egg white to be handled obtain ovalbumin, adopt combined-enzyme method again, obtain the albumin polypeptide product in conjunction with microwave method hydrolysed egg white protein.
The technical scheme that the present invention solves is as follows:
Get fresh hen egg, shell and get its egg white, slowly add isopyknic organic solvent deposit agent while stirring, 4 ℃ leave standstill 1-4h behind the stirring 30min, centrifugal, and the 4 ℃ of cold acetones that add two volumes down of supernatant liquor with obtaining fully stir, and are centrifugal.Merge the resulting throw out of two times centrifugal, this throw out is the ovalbumin crude product.Then with throw out according to adding distilled water at 1: 4, transfer to pH4.5-5.0, soaked at normal temperatures 1-2 hour, every 15min stirs once, back suction filtration, to make protein concn be 5% to adding distil water again, add prozyme, carry out the intermittent type heat treated with microwave, temperature is 45 ℃, PH4.5-5.0, the treatment time is 5min.Be warming up to 80 ℃ then, the 10min enzyme that goes out.The enzymolysis solution that will go out again behind the enzyme concentrates with diatomite filtration, vacuum-drying, and the product that obtains is the egg albumin polypeptide product.
The present invention adopts with the combinative enzyme hydrolysis ovalbumin, has increased the protein catalytic site, has overcome the incomplete shortcoming of single enzyme effect, effectively albumin is resolved into the low polypeptide of molecular weight.This reaction conditions gentleness combines combined-enzyme method and microwave technology, has not only shortened the reaction times, but also has following excellent part: 1, starting material egg material source is extensive, is food very common in people's daily life.2. the prozyme solution does not reduce the nutritive value of egg, can obtain the function of Duoing than egg protein.2, the polypeptide that obtains of enzymolysis can keep nutrition fully natural green attribute, does not contain any chemical substance, without any bitter taste and peculiar smell.3, the polypeptide that goes out of enzymolysis can not cause allergy, can increase important biological function and nutritive property that crude protein does not have.4, enzymolysis process can effectively be controlled the molecular weight section of polypeptide.5. microwave method is a kind of advanced person's a method, and it is simple to operate, good temperature-control performance, and the reaction times is short, the yield height of product.
The physiologic function of albumin polypeptide
Albumin polypeptide is as a kind of biologically active peptides, and it has its unique distinction aspect the physiological regulation of human body: 1. albumin polypeptide can be regulated body's immunity evenly.In the albumin polypeptide segment sequence among Ala-His, Val-His and the Val-His-Ala-Asp-Glu-Asu Histidine sequence can suppress the peroxidation of radical pair liposome (liposomas) phosphatidylcholine; protection immunocyte film reduces the cell peroxide level or eliminates the excessive free radicals that generates in the immune response pathway and influence body's immunological function.Add the material of polypeptide class in the food,, strengthen the energy that virus, bacterium are engulfed or killed and wounded to scavenger cell, natural killer cell etc., can improve the function of body opposing communicable disease by improving cellular immune function.Edible albumin polypeptide and nucleic acid can promote absorbing of Nucleotide, and the benefit that is of value to nucleic acid metabolism in the immunocyte asks synthetic, and animal body cellular immunization is had enhancement.2. can suppress Zinc metallopeptidase Zace1 (ACE) ovalbumin behind stomach en-and Quimotrase enzymolysis, obtain the biologically active peptides of several sequences, it can be by regulating RAAS system control hypertension, and the development of hypotensive agent is played crucial effect.3. catch free radical anti-oxidation stress, anti-ageing.Free radical is the product in the human body metabolic processes, along with the increase at age and the influence of disadvantageous internal and external environment (as radiation, smoking, environmental pollution, stress etc.), increasing sharply appears in the quantity of free radical, unsaturated fatty acids in the too much free radical attack cells film, in the chain type branched chain reaction of lipid peroxidation, constantly generate the lipid free radical, cell function day by day degenerate and fail, damage and pathology, so the toxic reaction of excessive free radical is the immediate cause that causes human senility and most of disease.At present by external removing free radical DPPH (2,21---trinitrophenyl-hydrazine) experiment show, albumin polypeptide has the effect of the free radical of removing anti-oxidation stress, mainly contain aromatic amino acid in its peptide chain that constitutes by 5～16 amino acid, form as amino acid such as the proline(Pro) of Xie Ansuan, leucine, N-terminal, Histidine, tyrosine.The mechanism that albumin polypeptide is caught the free radical anti-oxidation stress is: the sequence of the Asp～Arg-Val-Tyr of N one end provides _ " condition; His itself is reduced to 2 one different imdazole derivatives, so the Asp-Arg-Val-Tyr sequence is crucial to anti-oxidant activity to free radical for the His residue provides a hydrogen atom in a. peptide chain.B. imidazole ring is to the sequestering action of metal ion, and (the active inactivation of Fe, Cu ") catalysis free chain reaction reduces the generation of free radical to make metal ion.C. Histidine series can suppress the oxidation of radical pair liposome (liposomes) phosphatidylcholine, the normal function of protection cytolemma.4. adjusting gastrointestinal function.Gi tract are organs of digesting and assimilating of human nutrition material, and are very important to the health of human body.There is the microorganism that belongs to normal microflora in a large number in the human intestinal, body's immunity just often, various bacterial classifications suppress mutually, antagonism, keep the microecological balance in the enteron aisle, but under the state of some pathology (as gastro-enteritis, intestinal obstruction, surgical operation etc.), digestive tube lacks the effective stimulus of food, and the undersupply of nutrient substance causes the intestinal mucosa atrophy, and intestinal function and barrier are impaired.Also can cause alteration of intestinal flora, physiological anaerobism quantity descends, the conditioned pathogen abnormality proliferation.Have the scholar to report, glutamine (Gln) helps keeping structure, metabolism and function stable of mucous membrane of small intestine, and the effect that improves intestinal mucosal barrier has been widely used in clinical.This is because L-glutamic acid and glutamine two seed amino acids are being brought into play crucial effects in the metabolism of nitrogen and in amino acid whose oxidative metabolism, albumin polypeptide is for adjusting intestinal microflora, promote the rehabilitation of ultrashort bowel syndrome and the transformation efficiency that improves digestion food, promote the absorption of iron that obvious effect is all arranged, this is because albumin polypeptide is rich in L-glutamic acid and glutamine, its content is up to about 18%, and can provide essential amino acid and peptide chain for probiotic bacterium, therefore albumin polypeptide can be safeguarded intestinal microflora, promote the propagation of bifidus bacillus, thereby the adjusting gastrointestinal function has good nutritional support and assisting therapy effect for the crowd who has digestive tract diseases and get old and weak.
Albumin polypeptide is the natural product that obtains by enzyme engineering from ovalbumin, nontoxic, can take for a long time, the nutritive value that not only has egg, body essential amino acid and energy supply are provided, also have simultaneously and regulate immune, many-sided physiological regulation functions such as inhibition ACE is active and anti-oxidant, anti-ageing, adjusting gastrointestinal bacterial flora, having broad application prospects aspect the exploitation trophicity functional foodstuff.
Claims (4)
1. the preparation method of an egg albumin polypeptide is characterized in that may further comprise the steps:
Get fresh hen egg, shell and get its egg white, slowly add isopyknic organic solvent deposit agent while stirring, 4 ℃ leave standstill 1-4h behind the stirring 30min, centrifugal, and the 4 ℃ of cold acetones that add two volumes down of supernatant liquor with obtaining fully stir, and are centrifugal.Merge the resulting throw out of two times centrifugal, this throw out is the ovalbumin crude product.Then with throw out according to adding distilled water at 1: 4, transfer to pH4.5-5.0, soaked at normal temperatures 1-2 hour, every 15min stirs once, back suction filtration, to make protein concn be 5% to adding distil water again, add prozyme 7000 (U/g albumen), carry out the intermittent type heat treated with microwave, temperature is 45 ℃, PH4.5-5.0, the treatment time is 5min.Be warming up to 80 ℃ then, the 10min enzyme that goes out.The enzymolysis solution that will go out again behind the enzyme concentrates with diatomite filtration, vacuum-drying, and the product that obtains is the egg albumin polypeptide product.
2. organic solvent deposit agent according to claim 1 is characterized in that said: and 0.5mol/L Tricholroacetic Acid---acetone (1: 2, V/V) or ethanol---acetone (1: 1, V/V).
3. prozyme, it is characterized in that: neutral protease: papoid: bromeline=1: 2: 4, temperature are 45 ℃, PH4.5-5.0; Perhaps papoid: neutral protease: Sumizyme MP=2: 1: 2,55 ℃ of temperature, PH5.5-6.5.
4. according to the right 1 described the finished product egg albumin polypeptide that obtains, it is characterized in that this product is the mixture of albumin polypeptide molecule.
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| CN200810121543A CN101712974A (en) | 2008-10-07 | 2008-10-07 | Method for preparing egg albumin polypeptide |
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| CN200810121543A CN101712974A (en) | 2008-10-07 | 2008-10-07 | Method for preparing egg albumin polypeptide |
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102586376A (en) * | 2012-02-24 | 2012-07-18 | 大连工业大学 | Albumen antioxidant peptide and preparation method thereof |
| CN103146791A (en) * | 2013-03-13 | 2013-06-12 | 扬州大学 | Method for hydrolyzing egg-white proteins by various proteases |
| CN104988198A (en) * | 2015-07-22 | 2015-10-21 | 福建农林大学 | Preparation method of ACE inhibitory peptide through enzymatic hydrolysis of ovalbumin |
| CN105707407A (en) * | 2016-03-07 | 2016-06-29 | 湖北工业大学 | Preparing method of protein concentrate of sacha inchi fruits |
| CN106333276A (en) * | 2016-08-30 | 2017-01-18 | 滁州学院 | Novel dried egg and processing technology thereof |
| CN107047925A (en) * | 2017-05-02 | 2017-08-18 | 西北农林科技大学 | A kind of method of efficiently gentle reduction egg albumen sensitization |
| CN108864279A (en) * | 2018-07-24 | 2018-11-23 | 渤海大学 | A kind of ace inhibitory peptide |
| CN111254176A (en) * | 2020-01-20 | 2020-06-09 | 广东蛋能量生物科技有限公司 | Preparation method of egg white polypeptide |
| CN113214385A (en) * | 2021-05-26 | 2021-08-06 | 杭州佰倍优生物科技有限公司 | Enzymolysis ovalbumin composition for improving hypoalbuminemia and immunity |
| CN113337564A (en) * | 2021-06-18 | 2021-09-03 | 山东博瑞科生物技术有限责任公司 | Preparation method of efficient novel chicken egg albumin chelated zinc |
| CN113502206A (en) * | 2021-07-26 | 2021-10-15 | 吴艳 | Integrated production system of albumin peptide |
| CN114507279A (en) * | 2022-01-25 | 2022-05-17 | 湖北瑞邦生物科技有限公司 | Preparation method of antioxidant ovalbumin peptide |
-
2008
- 2008-10-07 CN CN200810121543A patent/CN101712974A/en active Pending
Cited By (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102586376A (en) * | 2012-02-24 | 2012-07-18 | 大连工业大学 | Albumen antioxidant peptide and preparation method thereof |
| CN102586376B (en) * | 2012-02-24 | 2013-10-23 | 大连工业大学 | Albumen antioxidant peptide and preparation method thereof |
| CN103146791A (en) * | 2013-03-13 | 2013-06-12 | 扬州大学 | Method for hydrolyzing egg-white proteins by various proteases |
| CN104988198A (en) * | 2015-07-22 | 2015-10-21 | 福建农林大学 | Preparation method of ACE inhibitory peptide through enzymatic hydrolysis of ovalbumin |
| CN105707407A (en) * | 2016-03-07 | 2016-06-29 | 湖北工业大学 | Preparing method of protein concentrate of sacha inchi fruits |
| CN105707407B (en) * | 2016-03-07 | 2019-10-25 | 湖北工业大学 | A kind of manufacturing method printing odd fruit protein concentrate |
| CN106333276B (en) * | 2016-08-30 | 2019-07-30 | 滁州学院 | A kind of dried eggs and its processing technology |
| CN106333276A (en) * | 2016-08-30 | 2017-01-18 | 滁州学院 | Novel dried egg and processing technology thereof |
| CN107047925A (en) * | 2017-05-02 | 2017-08-18 | 西北农林科技大学 | A kind of method of efficiently gentle reduction egg albumen sensitization |
| CN107047925B (en) * | 2017-05-02 | 2020-05-22 | 西北农林科技大学 | Method for efficiently and mildly reducing ovalbumin allergenicity |
| CN108864279A (en) * | 2018-07-24 | 2018-11-23 | 渤海大学 | A kind of ace inhibitory peptide |
| CN108864279B (en) * | 2018-07-24 | 2021-06-25 | 渤海大学 | ACE inhibitory peptide |
| CN111254176A (en) * | 2020-01-20 | 2020-06-09 | 广东蛋能量生物科技有限公司 | Preparation method of egg white polypeptide |
| CN113214385A (en) * | 2021-05-26 | 2021-08-06 | 杭州佰倍优生物科技有限公司 | Enzymolysis ovalbumin composition for improving hypoalbuminemia and immunity |
| CN113337564A (en) * | 2021-06-18 | 2021-09-03 | 山东博瑞科生物技术有限责任公司 | Preparation method of efficient novel chicken egg albumin chelated zinc |
| CN113502206A (en) * | 2021-07-26 | 2021-10-15 | 吴艳 | Integrated production system of albumin peptide |
| CN114507279A (en) * | 2022-01-25 | 2022-05-17 | 湖北瑞邦生物科技有限公司 | Preparation method of antioxidant ovalbumin peptide |
| CN114507279B (en) * | 2022-01-25 | 2023-11-21 | 湖北瑞邦生物科技有限公司 | Preparation method of antioxidant ovalbumin peptide |
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Open date: 20100526 |