CN110066778A - A kind of combined separation method of lysozyme from egg white and ovotransferrins - Google Patents
A kind of combined separation method of lysozyme from egg white and ovotransferrins Download PDFInfo
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- CN110066778A CN110066778A CN201910384796.8A CN201910384796A CN110066778A CN 110066778 A CN110066778 A CN 110066778A CN 201910384796 A CN201910384796 A CN 201910384796A CN 110066778 A CN110066778 A CN 110066778A
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- 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
- C07K14/79—Transferrins, e.g. lactoferrins, ovotransferrins
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2462—Lysozyme (3.2.1.17)
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- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01017—Lysozyme (3.2.1.17)
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Abstract
The invention discloses a kind of combined separation methods of lysozyme from egg white and ovotransferrins, belong to protein separation technical field, the method of the present invention includes that (1) egg white solution adjusts pH to 6.5-9.5, adsorbs lysozyme and ovotransferrins using metal-chelating resin cation；(2) eluent progress crystallization treatment is obtained into lysozyme crystal；(3) supernatant after crystallization is collected, is adsorbed using anion exchange resin, eluent is subjected to ultrafiltration and drying process, obtains ovotransferrins.Efficiently separating for lysozyme from egg white and ovotransferrins can be achieved at the same time in the method for the present invention, and yield is up to 75% or more, purity up to 95% or more, and used method has advantage easy to operate, at low cost, is easy to industrial application.
Description
Technical field
The present invention relates to a kind of combined separation methods of lysozyme from egg white and ovotransferrins, and it is pure to belong to Separation of Proteins
Change technical field.
Background technique
Albumen protein is optimal good protein in food, is mainly glued by ovalbumin, ovotransferrins, ovum
Albumen and lysozyme composition.Wherein lysozyme and ovotransferrins are the biological activity proteins paid close attention to by scientific research personnel, are had
Huge application potential.
Lysozyme is the essential antibacterial protein of infants growth and development, it can not only kill enteron aisle corruption coccus, but also can increase
The immune function of strong immunoglobulin, improves the anti-infection ability of baby, especially prevents weight loss, pre- to preemie
Anti- chylopoietic disease, enhancement weight and other effects, are the good auxiliary materials of baby's functional food.
Ovotransferrins is equally a kind of readily soluble non-crystallizable sugars albumen, passes through between molecular structure subregion and reversely puts down
Capable β chain is connected, this structure can be unfolded and be closed by hinge mechanism, is conducive to and the combination of ferric ion.Ovum
Transferrins by and ferric ion combination and deprived ferric ions necessary to many bacterial growths are bred, thus
Achieve the purpose that inhibit bacterial growth breeding, it is equal to pseudomonad, Escherichia coli and salmonella with broad-spectrum antibacterial
There is stronger inhibitory effect.The antibiotic property of ovotransferrins is similar to the lactoferrin of the mankind in quality in quantity, and disease-resistant
The performance of poison is even better than lactoferrin and serum transferrin, and in addition ovotransferrins also shows excellent anti-oxidant and anti-
Cancer activity.
But generally there is about the separation method of lysozyme and ovotransferrins low separation efficiency at present and be difficult to industry
The problem of change, therefore, develop it is a kind of it is easy, to efficiently separate lysozyme from egg white and the method for ovotransferrins be to have urgent city
Field demand.
Summary of the invention
In order to solve the problems, such as appeal, the invention proposes the friendships of metal-chelating cationic exchange resin adsorption-crystallization-anion
Resin sorption processes are changed, the lysozyme and ovotransferrins of high-purity can be obtained simultaneously, operating method is simply easy to industrialization, energy
It realizes utilization of resources, promotes the promotion of egg product enterprise added value.
The first purpose of the invention is to provide a kind of methods of Separation of Proteins in egg white, and the method includes walking as follows
It is rapid:
(1) egg white solution pH to 6.5-9.5 is adjusted, cation exchange resin is then added and ferric iron source carries out absorption point
From；Wherein the iron ion in ferric iron source is 0.05-0.4mmol/L with respect to the molar concentration of egg white solution；
(2) resin is eluted, eluent A is obtained；Adjust the pH of eluent A, concentration, crystallization obtain antalzyme crystallization and clear
Liquid B；
(3) pH of clear liquid B is adjusted, purification is concentrated to get ovotransferrins.
In one embodiment of the invention, cation exchange resin described in step (1) include carboxyl type it is weak sun from
Sub-exchange resin.
In one embodiment of the invention, ferric iron source described in step (1) includes iron chloride, ferric sulfate, nitric acid
The solubility trivalent iron salt such as iron.
In one embodiment of the invention, the mass ratio of resin and egg white solution is 1:(2-5 in step (1)).
In one embodiment of the invention, the preferred 1:3 of the mass ratio of resin and egg white solution in step (1).
It in one embodiment of the invention, further include cleaning resin before step (2) elution resin, the cleaning is benefit
It is cleaned with the NaCl aqueous solution of solid-liquid ratio 1:(2-5), wherein the mass concentration of NaCl aqueous solution is 0.5%-0.9%.
In one embodiment of the invention, the elution resin is using solid-liquid ratio 1:(2-5) NaCl solution into
Row elution, wherein the mass concentration of NaCl solution is 4%-7%.
In one embodiment of the invention, the pH of step (2) the eluent A is adjusted to 9.5-10.8.
In one embodiment of the invention, step (2) concentration, crystallization are that NaCl is added extremely into eluent A
Then 3%-9% mass concentration is concentrated, is placed at 5-10 DEG C and is crystallized.
In one embodiment of the invention, the pH of clear liquid B is adjusted to 6-8 in step (3).
In one embodiment of the invention, step (3) further include: when the pH to 6-8 of clear liquid B, according to solid-liquid ratio 1:
(2-5) is mixed with anion exchange resin, then removes supernatant, and resin is cleaned, is eluted, dries and turns iron to get ovum
Albumen.
In one embodiment of the invention, the method specifically comprises the following steps:
(1) egg liquid pre-processes: egg white solution stirring adjusts pH to 6.5-9.5；
(2) metal-chelating resin cation adsorbs: mixing resin and pretreated egg white solution according to solid-liquid ratio 1:5, adds
Enter 0.05-0.4mM ferric chloride solution, adsorbs 3-5h, remove supernatant A after the completion of absorption；
(3) it cleans: the NaCl solution that concentration is 0.9% is added into the resin for be completed absorption according to solid-liquid ratio 1:3, vibration
Supernatant, repeated washing 3-5 times are removed after swinging mixing；
(4) it elutes: the NaCl solution that concentration is 4-7%, oscillation elution is added according to solid-liquid ratio 1:3 in the resin of Xiang Qingxi
Clear liquid is collected after 30min, repetitive operation 3-5 times collects all eluent merging and is uniformly mixed so as to obtain eluent A；
(5) it crystallizes: eluent A being adjusted into pH to 9.5-10.8, NaCl is to salinity 3%-9% for addition, dense using ultrafiltration
Eluent is concentrated into protein concentration 2-5% by contracting technology, is stood at 10 DEG C and is formed crystallization, collects gained antalzyme crystallization respectively
With supernatant B；
(6) anion exchange resin adsorbs: taking supernatant B that pH value is adjusted to 6-8, hands over according to solid-liquid ratio 1:3 and anion
Resin mixing is changed, removes supernatant after adsorption reaction 3-5h；
(7) it elutes: after resin is cleaned with 0.9%NaCl solution, using concentration for the NaCl solution of 4-7%, oscillation is washed
Clear liquid is collected after de- 30min, repetitive operation 3-5 times collects all eluent merging and is uniformly mixed so as to obtain eluent；
(8) ultrafiltration and drying: carrying out ultrafiltration desalination for eluent, the ovotransferrins after obtained ultrafiltrate is dry i.e..
Second object of the present invention is applied to the above method in the purification of lysozyme from egg white.
Third object of the present invention is applied to the above method in egg white in the purification of ovotransferrins.
Fourth object of the present invention is that the above method is applied in egg white manufacture field.
Based on above-mentioned application, increase the added value of egg white, improves the availability of egg white.
The utility model has the advantages that
The method of the present invention can be achieved at the same time efficiently separating for lysozyme from egg white and ovotransferrins, yield up to 75% with
Upper, purity is up to 95% or more, and used method has advantage easy to operate, at low cost, is easy to industrialization and answers
With.
Detailed description of the invention
Fig. 1 is the separation method flow chart of lysozyme and ovotransferrins.
Specific embodiment
The present invention is based on multiple proteins in metal-chelating resin cation and crystallisation combined separation egg white, detailed processes
As shown in Figure 1.
Wherein pH range is 6.5-9.5 in step (1), to realize effective absorption of lysozyme and ovotransferrins, gained
It is rich in ovalbumin and ovomucin in supernatant A, is rich in lysozyme and ovotransferrins in precipitate B.
The resin type wherein selected in step (1) is metal-chelating resin cation, is a kind of carboxyl type weak cation
Exchanger resin, carboxyl can be acted on by metal-chelating to be realized and turn the absorption of iron to ovum, and is realized by electrostatic interaction molten
The absorption of bacterium enzyme.0.05-0.4mM ferric chloride solution is added during resin adsorption, adsorption time is 3-5 hours, to realize simultaneously
Effective absorption of lysozyme and ovotransferrins.
It is 4-7% that NaCl solution concentration used is eluted in step (2), elutes 30min, contains bacteriolyze in the eluent A of collection
Enzyme and ovotransferrins.
Crystallization condition described in step (2) be pH 9.5-10.8, protein concentration 2-5%, NaCl concentration 3%-9%, with
Guarantee that lysozyme effectively crystallizes precipitation.The uncrystallized lysozyme containing ovotransferrins and part in gained clear liquid B.
Anion exchange resin absorption is carried out to clear liquid B in step (3), adsorption conditions are pH 6-8, sorption reaction time
3-5h, to guarantee the effective of ovotransferrins, removing does not crystallize and unadsorbed lysozyme, elutes anion exchange resin, washes
De- conditional synchronization is rapid (2), is ovotransferrins in gained eluent B.
Purity of protein calculation method: isolated lysozyme and random transferrins are quantitatively divided using SDS-PAGE
It is pure up to albumen to calculate target protein ratio shared in total protein with gel imaging system to each protein band is acquired for analysis
Degree.
Albumen yield calculation method: it using the total content for the albumen that Kjeldahl's method test is isolated, is calculated according to purity
Target protein content m, after the same method in the egg white of calculation processing target protein total amount m0, according to formula albumen yield
=m/m0* 100 calculate the yield of target protein.
Embodiment 1:
(1) egg liquid pre-processes: egg white solution stirring adjusts pH to 8；
(2) metal-chelating resin cation adsorbs: locating according to solid-liquid ratio 1:5 by carboxyl type weak cation exchange resin and in advance
The egg white solution of reason mixes, and iron chloride is added, so that iron chloride molar concentration is 0.2mmol/L in egg white solution, adsorbs 5h, has adsorbed
Supernatant A is removed at moving back；
(3) it cleans: the NaCl solution that concentration is 0.9% is added into the resin for be completed absorption according to solid-liquid ratio 1:3, vibration
It swings and removes supernatant after mixing, repeated washing 5 times；
(4) it elutes: the NaCl solution that concentration is 5%, oscillation elution is added according to solid-liquid ratio 1:3 in the resin of Xiang Qingxi
Clear liquid is collected after 30min, repetitive operation 5 times, is collected all eluent merging and is uniformly mixed so as to obtain eluent A；
(5) it crystallizes: eluent A is adjusted into pH to 10, NaCl is added to salinity 5%, will be washed using ultrafiltration concentration technology
De- liquid is concentrated into protein concentration 3%, stands at 10 DEG C and forms crystallization, collects gained antalzyme crystallization and supernatant B respectively；
(6) anion exchange resin adsorbs: taking supernatant B that pH value is adjusted to 6, according to solid-liquid ratio 1:3 and anion exchange
Resin mixes, and removes supernatant after adsorption reaction 4h；
(7) it elutes: after resin is cleaned with 0.9%NaCl solution, using concentration for 5% NaCl solution, vibrate elution
Clear liquid is collected after 30min, repetitive operation 5 times, is collected all eluent merging and is uniformly mixed so as to obtain eluent B；
(8) ultrafiltration and drying: carrying out ultrafiltration desalination for eluent B, the ovotransferrins after obtained ultrafiltrate is dry i.e..
Gained lysozyme and ovotransferrins purity and yield calculated result are as shown in table 1:
The result of the separating obtained 2 kinds of protein of 1 embodiment of table 1
Embodiment 2:
(1) egg liquid pre-processes: egg white solution stirring adjusts pH to 9.5；
(2) metal-chelating resin cation adsorbs: locating according to solid-liquid ratio 1:5 by carboxyl type weak cation exchange resin and in advance
The egg white solution of reason mixes, and iron chloride is added, so that iron chloride molar concentration is 0.1mmol/L in egg white solution, adsorbs 5h, has adsorbed
Supernatant A is removed at moving back；
(3) it cleans: the NaCl solution that concentration is 0.9% is added into the resin for be completed absorption according to solid-liquid ratio 1:3, vibration
It swings and removes supernatant after mixing, repeated washing 4 times；
(4) it elutes: the NaCl solution that concentration is 7%, oscillation elution is added according to solid-liquid ratio 1:3 in the resin of Xiang Qingxi
Clear liquid is collected after 30min, repetitive operation 4 times, is collected all eluent merging and is uniformly mixed so as to obtain eluent A；
(5) it crystallizes: eluent A is adjusted into pH to 10.5, addition NaCl to salinity 7%, it will using the technology of ultrafiltration concentration
Eluent is concentrated into protein concentration 4%, stands at 10 DEG C and forms crystallization, collects gained antalzyme crystallization and supernatant B respectively；
(6) anion exchange resin adsorbs: taking supernatant B that pH value is adjusted to 7, according to solid-liquid ratio 1:3 and anion exchange
Resin mixes, and removes supernatant after adsorption reaction 5h；
(7) it elutes: after resin is cleaned with 0.9%NaCl solution, using concentration for 7% NaCl solution, vibrate elution
Clear liquid is collected after 30min, repetitive operation 5 times, is collected all eluent merging and is uniformly mixed so as to obtain eluent B；
(8) ultrafiltration and drying: carrying out ultrafiltration desalination for eluent B, the ovotransferrins after obtained ultrafiltrate is dry i.e..
Gained lysozyme and ovotransferrins purity and yield calculated result are as shown in table 2:
The result of the separating obtained 2 kinds of protein of 2 embodiment of table 2
Embodiment 3:
(1) egg liquid pre-processes: egg white solution stirring adjusts pH to 6.5；
(2) metal-chelating resin cation adsorbs: locating according to solid-liquid ratio 1:5 by carboxyl type weak cation exchange resin and in advance
The egg white solution of reason mixes, and iron chloride is added, so that iron chloride molar concentration is 0.2mmol/L in egg white solution, adsorbs 5h, has adsorbed
Supernatant A is removed at moving back；
(3) it cleans: the NaCl solution that concentration is 0.9% is added into the resin for be completed absorption according to solid-liquid ratio 1:3, vibration
It swings and removes supernatant after mixing, repeated washing 5 times；
(4) it elutes: the NaCl solution that concentration is 5%, oscillation elution is added according to solid-liquid ratio 1:3 in the resin of Xiang Qingxi
Clear liquid is collected after 30min, repetitive operation 5 times, is collected all eluent merging and is uniformly mixed so as to obtain eluent A；
(5) it crystallizes: eluent A is adjusted into pH to 10, NaCl is added to salinity 5%, will be washed using ultrafiltration concentration technology
De- liquid is concentrated into protein concentration 3%, stands at 10 DEG C and forms crystallization, collects gained antalzyme crystallization and supernatant B respectively；
(6) anion exchange resin adsorbs: taking supernatant B that pH value is adjusted to 6, according to solid-liquid ratio 1:3 and anion exchange
Resin mixes, and removes supernatant after adsorption reaction 4h；
(7) it elutes: after resin is cleaned with 0.9%NaCl solution, using concentration for 5% NaCl solution, vibrate elution
Clear liquid is collected after 30min, repetitive operation 5 times, is collected all eluent merging and is uniformly mixed so as to obtain eluent B；
(8) ultrafiltration and drying: carrying out ultrafiltration desalination for eluent B, the ovotransferrins after obtained ultrafiltrate is dry i.e..
Gained lysozyme and ovotransferrins purity and yield calculated result are as shown in table 3:
The result of the separating obtained 2 kinds of protein of 3 embodiment of table 3
Embodiment 4:
(1) egg liquid pre-processes: egg white solution stirring adjusts pH to 8；
(2) metal-chelating resin cation adsorbs: locating according to solid-liquid ratio 1:5 by carboxyl type weak cation exchange resin and in advance
The egg white solution of reason mixes, and iron chloride is added, so that iron chloride molar concentration is 0.05mmol/L in egg white solution, adsorbs 5h, absorption
Supernatant A is removed after the completion；
(3) it cleans: the NaCl solution that concentration is 0.9% is added into the resin for be completed absorption according to solid-liquid ratio 1:3, vibration
It swings and removes supernatant after mixing, repeated washing 5 times；
(4) it elutes: the NaCl solution that concentration is 5%, oscillation elution is added according to solid-liquid ratio 1:3 in the resin of Xiang Qingxi
Clear liquid is collected after 30min, repetitive operation 5 times, is collected all eluent merging and is uniformly mixed so as to obtain eluent A；
(5) it crystallizes: eluent A is adjusted into pH to 10, NaCl is added to salinity 5%, will be washed using ultrafiltration concentration technology
De- liquid is concentrated into protein concentration 3%, stands at 10 DEG C and forms crystallization, collects gained antalzyme crystallization and supernatant B respectively；
(6) anion exchange resin adsorbs: taking supernatant B that pH value is adjusted to 6, according to solid-liquid ratio 1:3 and anion exchange
Resin mixes, and removes supernatant after adsorption reaction 4h；
(7) it elutes: after resin is cleaned with 0.9%NaCl solution, using concentration for 5% NaCl solution, vibrate elution
Clear liquid is collected after 30min, repetitive operation 5 times, is collected all eluent merging and is uniformly mixed so as to obtain eluent B；
(8) ultrafiltration and drying: carrying out ultrafiltration desalination for eluent B, the ovotransferrins after obtained ultrafiltrate is dry i.e..
Gained lysozyme and ovotransferrins purity and yield calculated result are as shown in table 4:
The result of the separating obtained 2 kinds of protein of 4 embodiment of table 4
Embodiment 5:
(1) egg liquid pre-processes: egg white solution stirring adjusts pH to 8；
(2) metal-chelating resin cation adsorbs: locating according to solid-liquid ratio 1:5 by carboxyl type weak cation exchange resin and in advance
The egg white solution of reason mixes, and iron chloride is added, so that iron chloride molar concentration is 0.4mmol/L in egg white solution, adsorbs 5h, has adsorbed
Supernatant A is removed at moving back；
(3) it cleans: the NaCl solution that concentration is 0.9% is added into the resin for be completed absorption according to solid-liquid ratio 1:3, vibration
It swings and removes supernatant after mixing, repeated washing 5 times；
(4) it elutes: the NaCl solution that concentration is 5%, oscillation elution is added according to solid-liquid ratio 1:3 in the resin of Xiang Qingxi
Clear liquid is collected after 30min, repetitive operation 5 times, is collected all eluent merging and is uniformly mixed so as to obtain eluent A；
(5) it crystallizes: eluent A is adjusted into pH to 10, NaCl is added to salinity 5%, will be washed using ultrafiltration concentration technology
De- liquid is concentrated into protein concentration 3%, stands at 10 DEG C and forms crystallization, collects gained antalzyme crystallization and supernatant B respectively；
(6) anion exchange resin adsorbs: taking supernatant B that pH value is adjusted to 6, according to solid-liquid ratio 1:3 and anion exchange
Resin mixes, and removes supernatant after adsorption reaction 4h；
(7) it elutes: after resin is cleaned with 0.9%NaCl solution, using concentration for 5% NaCl solution, vibrate elution
Clear liquid is collected after 30min, repetitive operation 5 times, is collected all eluent merging and is uniformly mixed so as to obtain eluent B；
(8) ultrafiltration and drying: carrying out ultrafiltration desalination for eluent B, the ovotransferrins after obtained ultrafiltrate is dry i.e..
Gained lysozyme and ovotransferrins purity and yield calculated result are as shown in table 5:
The result of the separating obtained 2 kinds of protein of 5 embodiment of table 5
Reference examples 1:
Referring to embodiment 1, the molar concentration of the ferric trichloride in step (2) is replaced with into 0.02mM, other conditions are constant,
Multiple protein in separating treatment egg white, the results are shown in Table 6.
The result of the separating obtained 3 kinds of protein of 6 reference examples of table 1
Reference examples 2:
Referring to embodiment 1, the pH in step (1) is replaced with 5,10 respectively, other conditions are constant, in separating treatment egg white
Multiple protein, the results are shown in Table 7.
The result of the separating obtained 2 kinds of protein of 7 reference examples of table 2
Claims (10)
1. a kind of method for separating lysozyme from egg white and ovotransferrins, which is characterized in that described method includes following steps:
(1) egg white solution pH to 6.5-9.5 is adjusted, cation exchange resin is then added and ferric iron source carries out adsorbing separation；Its
Iron ion in middle ferric iron source is 0.05-0.4mmol/L with respect to the molar concentration of egg white solution；
(2) resin is eluted, eluent A is obtained；The pH of eluent A is adjusted, concentration, crystallization obtain antalzyme crystallization and clear liquid B；
(3) pH of clear liquid B is adjusted, purification is concentrated to get ovotransferrins.
2. the method according to claim 1, wherein the mass ratio of resin and egg white solution is 1:(2- in step (1)
5)。
3. method according to claim 1 or 2, which is characterized in that the ferric iron source includes iron chloride, ferric sulfate, nitre
One of sour iron is a variety of.
4. method according to claim 1 to 3, which is characterized in that the elution resin is to utilize solid-liquid ratio 1:(2-
5) NaCl aqueous solution is eluted, and wherein the mass concentration of NaCl aqueous solution is 4%-7%.
5. method according to claim 1 to 4, which is characterized in that the pH of step (2) the eluent A is adjusted to
9.5-10.8。
6. -5 any method according to claim 1, which is characterized in that step (2) described condensing crystallizing is to eluent A
Middle addition NaCl is concentrated to 3%-9% mass concentration, is subsequently placed at 5-10 DEG C and is crystallized.
7. -6 any method according to claim 1, which is characterized in that the pH of clear liquid B is adjusted to 6-8 in step (3).
8. application of any the method for claim 1-7 in egg white manufacture field.
9. application of any the method for claim 1-7 in purification lysozyme from egg white.
10. application of any the method for claim 1-7 in purification egg white in ovotransferrins.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103160482A (en) * | 2013-03-14 | 2013-06-19 | 江苏大学 | Method for preparing egg white lysozyme and active protein by adopting coseparation |
| CN108118044A (en) * | 2018-02-02 | 2018-06-05 | 华南理工大学 | A kind of method of separating-purifying egg white lysozyme |
-
2019
- 2019-05-09 CN CN201910384796.8A patent/CN110066778B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103160482A (en) * | 2013-03-14 | 2013-06-19 | 江苏大学 | Method for preparing egg white lysozyme and active protein by adopting coseparation |
| CN108118044A (en) * | 2018-02-02 | 2018-06-05 | 华南理工大学 | A kind of method of separating-purifying egg white lysozyme |
Non-Patent Citations (3)
| Title |
|---|
| A. AL-MASHDCHI AND SHURYO NAKAI: "Separation of Ovotransferrin from Egg White by", 《AGRIC. BIOL. CHEM.》 * |
| 傅冰等: "鸡蛋清中3种蛋白质的连续化提取工艺初步亚牛", 《广东农业科学》 * |
| 张亚辉等: "纤维素金属螯合亲和膜用于蛋清中", 《食品与生物技术学报》 * |
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