CN111153986A - Method for extracting ovotransferrin from pigeon eggs - Google Patents
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Abstract
The invention discloses a method for extracting ovotransferrin from pigeon eggs. The method comprises the following steps: (1) separating egg white from pigeon eggs, diluting the egg white, fully stirring, centrifuging, filtering, and taking supernatant; (2) then FeCl is added into the supernatant3·6H2Continuously and fully stirring O, sodium bicarbonate and sodium chloride; (3) adding ethanol into the supernatant to enable the volume concentration of the ethanol in the solution to reach 40-45%, uniformly stirring, standing, and centrifuging to obtain precipitate 1 and filtrate; (4) and continuously adding ethanol into the filtrate to ensure that the volume concentration of the ethanol in the filtrate reaches 55-65%, uniformly stirring, standing, and centrifuging to obtain a precipitate 2. (5) Mixing the precipitate 1 and the precipitate 2, washing with ethanol, adding water for redissolution, mixing with resin, stirring, vacuum filtering, dialyzing, and lyophilizing to obtain the ovotransferrinA protein. The invention provides a brand-new method for extracting ovotransferrin from pigeon eggs, and the method has high extraction rate and purity.
Description
Technical Field
The invention relates to the technical field of ovotransferrin preparation, in particular to a method for extracting ovotransferrin from pigeon eggs.
Background
Ovotransferrin, also known as conalbumin, is an important single-chain glycoprotein in avian egg white, accounting for about 12% of the total protein content of egg white. The protein molecule consists of 686 amino acid residues, has a molecular weight range of 78-80 kDa, and can be combined with metal ions such as iron, copper or zinc to greatly change the conformation of the ovotransferrin. Ovotransferrin has various physiological functions, such as the function of enhancing the immunity of the organism, can promote the growth of lymphocytes and prevent or inhibit the invasion of pathogenic bacteria. The ovotransferrin can also transport iron ions to parts of the body which need iron, thereby promoting the absorption of the body to iron and nutrient substances and effectively preventing anemia. Ovotransferrin also has excellent antibacterial function, especially against intestinal pathogens and viruses, and can be used for treating acute enteritis in infants. In addition, ovotransferrin can inhibit the generation of free radicals, has an antioxidant effect, and has certain effects of relieving rheumatoid arthritis and preventing aging. Therefore, the application prospect of the ovotransferrin is very wide, for example, the ovotransferrin can be used as a functional component to be added into food, or can be prepared into natural preservatives or bacteriostats, or can be prepared into medical products and the like.
Pigeon egg refers to egg of original pigeon or domestic pigeon of dove family, and is called "animal ginseng". Pigeon egg has effects of improving skin cell activity and elasticity fiber in skin, improving blood circulation, clearing away heat and toxic materials, and preventing children measles. The pigeon eggs contain a large amount of high-quality protein, a small amount of fat, a small amount of sugar, phospholipid, iron, calcium, vitamin A, vitamin B1, vitamin D and other nutritional ingredients, and are easy to digest and absorb. Compared with eggs, the pigeon eggs have higher riboflavin content, lecithin content, calcium, iron element and collagen content than eggs and lower fat content, so the pigeon eggs are very suitable for patients with hyperlipidemia, women and infants. At present, the research on the nutritional value of pigeon eggs is more and more, so the research on the protein in the pigeon eggs has important significance.
At present, methods for extracting ovotransferrin mainly comprise an alcohol precipitation method, a salt precipitation method, an anion-cation resin exchange method, a membrane separation technical method and the like. However, the existing extraction method has the problems of low extraction rate and low purity; therefore, the large-scale industrial production extraction of the ovotransferrin is limited. In addition, no report is available for extracting ovotransferrin from pigeon eggs.
Disclosure of Invention
The technical problem to be solved by the invention is to provide a method for extracting ovotransferrin from pigeon eggs in order to overcome the technical defect of lacking of extracting ovotransferrin from pigeon eggs in the prior art; the method can obtain high extraction amount of ovotransferrin.
The technical problem to be solved by the invention is realized by the following technical scheme:
a method for extracting ovotransferrin from pigeon eggs, which is characterized by comprising the following steps:
(1) separating egg white from pigeon eggs, diluting the egg white, fully stirring, centrifuging, filtering, and taking supernatant;
(2) then FeCl is added into the supernatant3·6H2Continuously and fully stirring O, sodium bicarbonate and sodium chloride;
(3) adding ethanol into the supernatant to enable the volume concentration of the ethanol in the solution to reach 40-45%, uniformly stirring, standing, and centrifuging to obtain precipitate 1 and filtrate;
(4) and continuously adding ethanol into the filtrate to ensure that the volume concentration of the ethanol in the filtrate reaches 55-65%, uniformly stirring, standing, and centrifuging to obtain a precipitate 2.
(5) And combining the precipitate 1 and the precipitate 2, washing with ethanol, adding water for redissolution, mixing with resin, stirring, carrying out suction filtration, dialysis and freeze drying to obtain the ovotransferrin.
Preferably, the step (1) of diluting the egg white refers to diluting the egg white with water; the mass consumption of the water is 1.5-3 times of the mass of the egg white liquid.
Most preferably, the amount of water is 2 times the mass of the egg white.
Preferably, the stirring in the step (1) adopts magnetic stirring; the stirring time is 50-70 min.
Most preferably, the stirring in step (1) is magnetic stirring; the stirring time was 60 min.
Preferably, the centrifugation in the step (1) is performed at 3-5 ℃, the centrifugation speed is 5500-6500 r/min, and the centrifugation time is 15-25 min.
Most preferably, the centrifugation in step (1) means centrifugation at 4 ℃ at 6000r/min for 20 min.
Preferably, FeCl is used in step (2)3·6H2The mass ratio of the O, the sodium bicarbonate and the sodium chloride to the egg white is 3-3.5: 2.1-2.6: 5-6: 200.
most preferably, FeCl in step (2)3·6H2The mass ratio of the O, the sodium bicarbonate and the sodium chloride to the egg white is 3.24: 2.52: 5.256: 200.
preferably, magnetic stirring is adopted for stirring in the step (2); the stirring time is 100-150 min.
Most preferably, the stirring in the step (2) adopts magnetic stirring; the stirring time was 120 min.
Preferably, the ethanol volume concentration in step (3) reaches 43%.
Preferably, the ethanol volume concentration in step (4) reaches 59%.
Through a large amount of experimental researches, the inventor finds that the dilution times, the stirring time, the centrifugal speed and the centrifugal time in the step (1) and the FeCl in the step (2)3·6H2The mass and dosage of O, sodium bicarbonate and sodium chloride, the stirring time, the volume concentration of the ethanol in the steps (3) and (4) and other factors play an important role in the extraction of a large amount of ovotransferrin from pigeon eggs; changes in each factor affect the amount of ovotransferrin extracted. The research of the inventor shows that the higher extraction amount of the ovotransferrin can be obtained only if all factors are under the condition of the invention.
Preferably, the ovotransferrin obtained in the step (5) is further ultrafiltered to obtain the ovotransferrin with higher purity.
Most preferably, the ultrafiltration is performed by using a filter membrane with the diameter of 50mm and the diameter of 0.45 mu m; the ultrafiltration temperature was 25 ℃ and the pressure was 5 psi.
Has the advantages that: the invention provides a brand new method for extracting ovotransferrin from pigeon eggs, which has higher extraction rate and can also extract high-purity ovotransferrin from the pigeon eggs. The invention provides an optimized scheme for large-scale production and extraction of ovotransferrin, and has certain significance for deeply researching the nutritional value of pigeon eggs.
Drawings
Figure 1 is an electrophoretic picture of ovotransferrin extracted from pigeon eggs.
Figure 2 is a line graph of the hydroxyl radical scavenging capacity of different concentrations of ovotransferrin and Vc.
Figure 3 is a line graph of the reduction capacity of iron ions by different concentrations of ovotransferrin and Vc.
FIG. 4 is a line graph of the scavenging capacity of different concentrations of ovotransferrin and Vc for superoxide anion free radicals.
Figure 5 is a line graph of the free radical scavenging capacity of DPPH against ovotransferrin and Vc at different concentrations.
Detailed Description
The present invention is further explained below with reference to specific examples, which are not intended to limit the present invention in any way.
Example 1 method for extracting ovotransferrin from Pigeon eggs
(1) Separating 200g egg white from pigeon eggs, adding 400g water into the egg white for dilution, magnetically stirring for 60min, centrifuging at 4 deg.C and 6000r/min for 20min, filtering with gauze, and collecting supernatant;
(2) then FeCl is added into the supernatant3·6H2O3.24 g, sodium bicarbonate 2.52g and sodium chloride 5.256g, and then stirring for 120min by magnetic force;
(3) adding absolute ethanol into the supernatant to make the volume concentration of ethanol in the supernatant reach 43%, stirring, homogenizing, standing for 30min, and centrifuging to obtain precipitate 1 and filtrate;
(4) and (3) continuously adding absolute ethyl alcohol into the filtrate to ensure that the volume concentration of the ethyl alcohol in the filtrate reaches 59%, uniformly stirring, standing overnight, and centrifuging to obtain a precipitate 2.
(5) Mixing the precipitate 1 and the precipitate 2, washing with anhydrous ethanol, re-dissolving the precipitate with 10 times of distilled water, and mixing with cationic resin (1/3 volume of distilled water) under stirring for 30 min; and carrying out suction filtration, dialysis (by adopting a dialysis bag or a dialysis column of 70 kda) and freeze-drying to obtain 3.2g of the ovotransferrin.
Example 2 method for extracting ovotransferrin from Pigeon eggs
(1) Separating egg white liquid 200g from pigeon egg, adding 300g water into egg white liquid for dilution, magnetically stirring for 70min, centrifuging at 5500r/min at 4 deg.C for 25min, filtering with gauze, and collecting supernatant;
(2) then FeCl is added into the supernatant3·6H2O3.24 g, sodium bicarbonate 2.52g and sodium chloride 5.256g, and then stirring for 100min by magnetic force;
(3) adding absolute ethanol into the supernatant to make the volume concentration of ethanol in the supernatant reach 40%, stirring, homogenizing, standing for 30min, and centrifuging to obtain precipitate 1 and filtrate;
(4) and (3) continuously adding absolute ethyl alcohol into the filtrate to enable the volume concentration of the ethyl alcohol in the filtrate to reach 65%, uniformly stirring, standing overnight, and centrifuging to obtain a precipitate 2.
(5) Mixing the precipitate 1 and the precipitate 2, washing with anhydrous ethanol, re-dissolving the precipitate with 10 times of distilled water, and mixing with cationic resin (1/3 volume of distilled water) under stirring for 30 min; and carrying out suction filtration, dialysis (by adopting a dialysis bag or a dialysis column of 70 kda) and freeze-drying to obtain 2.3g of the ovotransferrin.
Example 3 method for extracting ovotransferrin from Pigeon eggs
(1) Separating 200g egg white from pigeon eggs, adding 600g water into the egg white for dilution, magnetically stirring for 50min, centrifuging at 4 ℃ for 15min at 6500r/min, filtering with gauze, and collecting supernatant;
(2) then FeCl is added into the supernatant3·6H2O3.24 g, sodium bicarbonate 2.52g and sodium chloride 5.256g, and then stirring for 150min by magnetic force;
(3) adding absolute ethanol into the supernatant to make the volume concentration of ethanol in the supernatant reach 45%, stirring, homogenizing, standing for 30min, and centrifuging to obtain precipitate 1 and filtrate;
(4) and (3) continuously adding absolute ethyl alcohol into the filtrate to ensure that the volume concentration of the ethyl alcohol in the filtrate reaches 55%, uniformly stirring, standing overnight, and centrifuging to obtain a precipitate 2.
(5) Mixing the precipitate 1 and the precipitate 2, washing with anhydrous ethanol, re-dissolving the precipitate with 10 times of distilled water, and mixing with cationic resin (1/3 volume of distilled water) under stirring for 30 min; and carrying out suction filtration, dialysis (by adopting a dialysis bag or a dialysis column of 70 kda) and freeze-drying to obtain 2.5g of the ovotransferrin.
Comparative example 1 method for extracting ovotransferrin from pigeon eggs
To illustrate the dilution factor, stirring time, centrifugation speed and time in step (1), FeCl in step (2)3·6H2The mass and dosage of O, sodium bicarbonate and sodium chloride, the stirring time, the volume concentration of the ethanol in the steps (3) and (4) and other factors influence the extraction amount of the ovotransferrin in the pigeon eggs.
The comparative example changes the dilution times, stirring time, centrifugal speed and time in the step (1); in the comparative example, 800g of water is added into the egg white liquid for dilution, magnetic stirring is carried out for 90min, and then centrifugation is carried out for 40min at 7000r/min at 4 ℃; the rest of the conditions were the same as in example 1; this comparative example gives 1.9g of the ovotransferrin described. The extraction of ovotransferrin obtained in this comparative example is much less than 3.2g of example 1.
Comparative example 2 method for extracting ovotransferrin from pigeon eggs
To illustrate the dilution factor, stirring time, centrifugation speed and time in step (1), FeCl in step (2)3·6H2The mass and dosage of O, sodium bicarbonate and sodium chloride, the stirring time, the volume concentration of the ethanol in the steps (3) and (4) and other factors influence the extraction amount of the ovotransferrin in the pigeon eggs.
This comparative example changed FeCl in step (2)3·6H2The mass dosages of O, sodium bicarbonate and sodium chloride; comparative example FeCl was added to the supernatant3·6H26.48g of O, 1.95g of sodium bicarbonate and 3.942g of sodium chloride; the rest of the conditions were the same as in example 1; this comparative example gives 1.8g of the ovotransferrin described. The extraction of ovotransferrin obtained in this comparative example is much less than 3.2g of example 1.
Comparative example 3 method for extracting ovotransferrin from pigeon eggs
To illustrate the dilution factor, stirring time, centrifugation speed and time in step (1), FeCl in step (2)3·6H2The mass and dosage of O, sodium bicarbonate and sodium chloride, the stirring time, the volume concentration of the ethanol in the steps (3) and (4) and other factors influence the extraction amount of the ovotransferrin in the pigeon eggs.
This comparative example changed FeCl in step (2)3·6H2The mass amounts of O, sodium bicarbonate and sodium chloride and the stirring time; comparative example FeCl was added to the supernatant3·6H26.48g of O, 1.95g of sodium bicarbonate and 3.942g of sodium chloride, and then stirring for 60min by magnetic force; the rest of the conditions were the same as in example 1; this comparative example gives 1.6g of the ovotransferrin described. The extraction of ovotransferrin obtained in this comparative example is much less than 3.2g of example 1.
Comparative example 4 method for extracting ovotransferrin from pigeon eggs
To illustrate the dilution factor, stirring time, centrifugation speed and time in step (1), FeCl in step (2)3·6H2O, hydrogen carbonateThe mass usage amount of sodium and sodium chloride, the stirring time, the volume concentration of ethanol in the steps (3) and (4) and other factors influence the extraction amount of the ovotransferrin in the pigeon eggs.
This comparative example varied the volume concentration of ethanol in steps (3) and (4); in the step (3) of the comparative example, absolute ethyl alcohol is added into the supernatant, so that the volume concentration of the ethyl alcohol in the supernatant reaches 50 percent; step (4), adding absolute ethyl alcohol into the filtrate continuously to enable the volume concentration of the ethyl alcohol in the filtrate to reach 50%; the rest of the conditions were the same as in example 1; this comparative example gives 1.7g of the ovotransferrin described. The extraction of ovotransferrin obtained in this comparative example is much less than 3.2g of example 1.
As can be seen from comparative examples 1 to 4, the dilution factor, stirring time, centrifugation speed and time in step (1), and FeCl in step (2)3·6H2The mass and dosage of O, sodium bicarbonate and sodium chloride, the stirring time, the volume concentration of the ethanol in the steps (3) and (4) and other factors influence the extraction amount of the ovotransferrin in the pigeon eggs. All factors can obtain higher extraction amount of ovotransferrin only under the comprehensive conditions of the invention.
Example 4 method for extracting ovotransferrin from Pigeon eggs
Further carrying out ultrafiltration on the ovotransferrin extracted in the embodiment 1 to obtain the ovotransferrin with higher purity; the ultrafiltration adopts a filter membrane with the diameter of 50mm and the diameter of 0.45 mu m; the ultrafiltration temperature was 25 ℃ and the pressure was 5 psi. Comparing the result of the electrophoresis of the ovotransferrin obtained in the example with that of pure ovotransferrin, wherein the result is shown in fig. 1, the rightmost side of the graph 1 is an electrophoretogram of pure ovotransferrin, and the leftmost side is an electrophoretogram of ovotransferrin prepared in the example; as can be seen from FIG. 1, the ovotransferrin of the example is enriched and the impurity protein is less, so that the ovotransferrin with higher purity can be extracted.
Experimental example measurement of oxidation resistance of ovotransferrin
Ovotransferrin (hereinafter referred to as a sample) prepared in example 1 was subjected to a study of oxidation resistance. The hydroxyl radical scavenging capacity, the iron ion reducing capacity, the superoxide anion radical scavenging capacity and the DPPH radical scavenging capacity are characterized by 4 indexes and compared with the same concentration Vc. The method comprises the following specific steps:
1. hydroxyl radical scavenging ability:
0.2, 0.4, 0.6, 0.8 and 1.0mg/mL of sample are respectively added with 1mL of 9mM ferrous sulfate, 1mL of 9Mm salicylic acid-ethanol solution and 1mL of 9mM hydrogen peroxide in sequence to easily start reaction, after oscillation and uniform mixing, the mixture is put into a water bath at 37 ℃ for 30min, the distilled water is used as a blank, and the A is measured in 510nm ultraviolet light1Using distilled water instead of hydrogen peroxide, test A2Measuring A by using distilled water instead of protein0The control group had the same concentration of Vc.
As shown in figure 2, the clearance capacity of the ovotransferrin to hydroxyl radicals is gradually enhanced with the increase of the concentration of the ovotransferrin, but the clearance capacity of the ovotransferrin to the hydroxyl radicals is lower than that of the ovotransferrin at the same concentration Vc.
2. Iron ion reducing ability:
0.2, 0.4, 0.6, 0.8 and 1.0mg/mL of each sample is added with 2mL of 0.2mol/L of phosphoric acid buffer (pH is 6.6) and 2mL of 1% potassium ferricyanide solution, after mixing, the mixture is put in a water bath at 50 ℃ for 20min, 2mL of 10% trichloroacetic acid solution is added, after shaking and mixing, the mixture is centrifuged at 3000r/min for 10min, supernatant is kept, 2mL of supernatant is taken, 2mL of distilled water and 0.4mL of 0.1% ferric trichloride solution are added, after mixing, the mixture is kept at 50 ℃ for 10min, when the solution turns from yellow to blue, the absorbance is measured at 700nm, the distilled water is used as a blank control instead of the sample, and the control group is vitamin C with the same concentration.
As shown in figure 3, the reducing capacity of the ovotransferrin to iron ions is gradually enhanced with the increase of the ovotransferrin concentration, but the reducing capacity of the ovotransferrin to the iron ions is weaker than that of the ovotransferrin to the iron ions at the same concentration Vc
3. Determination of superoxide anion radical scavenging ability:
4mL of sample solutions with different concentrations respectively, using distilled water as blank control, adding 4.5mL and 0.1mol/L Tris-HCl buffer solution pH8.2 into each tube, shaking and mixing uniformly, carrying out water bath at 25 ℃ for 20min, adding 0.3mL3mM pyrogallol respectively, measuring the absorbance value at 320nm wavelength immediately after mixing uniformly, recording once every 30s, timing for 4min, adjusting zero by using the blank control to obtain a regression equation of absorbance along with time change, wherein the control group is Vc with the same concentration
As shown in figure 4, the scavenging capacity of the ovotransferrin to superoxide anion free radicals is gradually enhanced along with the increase of the concentration of the ovotransferrin, but the scavenging capacity of the ovotransferrin to superoxide anion free radicals is lower than that of the ovotransferrin with the same concentration Vc.
DPPH radical scavenging ability
2mL of 2mM DPPH solution (95% ethanol) was added to each sample, and the mixture was shaken well, with 95% ethanol being used as a sample control instead of DPPH, and 2mL of distilled water being used as a blank control instead of DPPH, and absorbance was measured at 517nm for 3 replicates. The control group is Vc solution with the same concentration.
As shown in figure 5, the clearance capacity of the ovotransferrin to DPPH free radicals is gradually enhanced along with the increase of the concentration of the ovotransferrin, but the clearance capacity of the ovotransferrin to DPPH free radicals is lower than that of the ovotransferrin with the same concentration Vc.
Claims (10)
1. A method for extracting ovotransferrin from pigeon eggs, which is characterized by comprising the following steps:
(1) separating egg white from pigeon eggs, diluting the egg white, fully stirring, centrifuging, filtering, and taking supernatant;
(2) then FeCl is added into the supernatant3·6H2Continuously and fully stirring O, sodium bicarbonate and sodium chloride;
(3) adding ethanol into the supernatant to enable the volume concentration of the ethanol in the solution to reach 40-45%, uniformly stirring, standing, and centrifuging to obtain precipitate 1 and filtrate;
(4) and continuously adding ethanol into the filtrate to ensure that the volume concentration of the ethanol in the filtrate reaches 55-65%, uniformly stirring, standing, and centrifuging to obtain a precipitate 2.
(5) And combining the precipitate 1 and the precipitate 2, washing with ethanol, adding water for redissolution, mixing with resin, stirring, carrying out suction filtration, dialysis and freeze drying to obtain the ovotransferrin.
2. The method for extracting ovotransferrin from pigeon eggs according to claim 1, wherein the step of diluting the egg white in step (1) is to dilute the egg white with water; the mass consumption of the water is 1.5-3 times of the mass of the egg white liquid; most preferably, the amount of water is 2 times the mass of the egg white.
3. The method for extracting ovotransferrin from pigeon eggs according to claim 1, wherein the stirring in step (1) is magnetic stirring; stirring for 50-70 min; most preferably, the stirring in step (1) is magnetic stirring; the stirring time was 60 min.
4. The method for extracting ovotransferrin from pigeon eggs according to claim 1, wherein the centrifugation in step (1) is carried out at 3-5 ℃, the centrifugation speed is 5500-6500 r/min, and the centrifugation time is 15-25 min; most preferably, the centrifugation in step (1) means centrifugation at 4 ℃ at 6000r/min for 20 min.
5. The method for extracting ovotransferrin from pigeon eggs according to claim 1, wherein FeCl is used in step (2)3·6H2The mass ratio of the O, the sodium bicarbonate and the sodium chloride to the egg white is 3-3.5: 2.1-2.6: 5-6: 200 of a carrier; most preferably, FeCl in step (2)3·6H2The mass ratio of the O, the sodium bicarbonate and the sodium chloride to the egg white is 3.24: 2.52: 5.256: 200.
6. the method for extracting ovotransferrin from pigeon eggs according to claim 1, wherein the stirring in step (2) is magnetic stirring; stirring for 100-150 min; most preferably, the stirring in the step (2) adopts magnetic stirring; the stirring time was 120 min.
7. The method for extracting ovotransferrin from pigeon eggs according to claim 1, wherein the volume concentration of ethanol in step (3) is up to 43%.
8. The method for extracting ovotransferrin from pigeon eggs according to claim 1, wherein the volume concentration of ethanol in step (4) is up to 59%.
9. The method of claim 1, wherein the ovotransferrin obtained from step (5) is further ultrafiltered to obtain a more pure ovotransferrin; the ultrafiltration adopts a filter membrane with the diameter of 50mm and the diameter of 0.45 mu m; the ultrafiltration temperature was 25 ℃ and the pressure was 5 psi.
10. Ovotransferrin obtainable by a process according to any one of claims 1 to 9.
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|---|---|---|---|---|
| CN112028990A (en) * | 2020-09-08 | 2020-12-04 | 兴宁市金鸽养殖专业合作社 | Extraction method of pigeon egg white ovalbumin |
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| CN102796193A (en) * | 2012-07-23 | 2012-11-28 | 华中农业大学 | Method for extracting ovotransferrin from egg white |
| CN102875669A (en) * | 2012-10-29 | 2013-01-16 | 天津商业大学 | Method for separating and extracting ovotransferrin |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102796193A (en) * | 2012-07-23 | 2012-11-28 | 华中农业大学 | Method for extracting ovotransferrin from egg white |
| CN102875669A (en) * | 2012-10-29 | 2013-01-16 | 天津商业大学 | Method for separating and extracting ovotransferrin |
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| Title |
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| NORIKO SUZUKI，ET AL: "Isolation and Characterization of Major Glycoproteins of Pigeon Egg White", 《THE JOURNAL OF BIOLOGICAL CHEMISTRY》 * |
| 佟永薇: "卵转铁蛋白的性质及分离纯化研究", 《万方数据》 * |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112028990A (en) * | 2020-09-08 | 2020-12-04 | 兴宁市金鸽养殖专业合作社 | Extraction method of pigeon egg white ovalbumin |
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