CN103444979A - Method for lowering albumen allergenicity by immobilized enzyme hydrolysis - Google Patents
Method for lowering albumen allergenicity by immobilized enzyme hydrolysis Download PDFInfo
- Publication number
- CN103444979A CN103444979A CN2013103453475A CN201310345347A CN103444979A CN 103444979 A CN103444979 A CN 103444979A CN 2013103453475 A CN2013103453475 A CN 2013103453475A CN 201310345347 A CN201310345347 A CN 201310345347A CN 103444979 A CN103444979 A CN 103444979A
- Authority
- CN
- China
- Prior art keywords
- magnetic bead
- albumen
- reaction
- buffer solution
- pbs buffer
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to a method for lowering albumen allergenicity by immobilized enzyme hydrolysis, which comprises the following steps: immobilizing crosslinking proteinase with magnetic beads, hydrolyzing albumen with the immobilized proteinase, removing fishy smell from the hydrolysate by using activated carbon, carrying out vacuum concentration at 60-70 DEG C, and carrying out freeze-drying to obtain the albumen powder of which the water content is less than 5%. The albumen powder has low allergenicity, contains abundant peptides and amino acids required by the human body, can be easily absorbed by the human body, and does not have bad flavor; and the method has favorable popularization and application prospects, can be widely used in the fields of food, drugs and the like, and is especially suitable to be used as a food raw material for allergic patients.
Description
Technical field
The invention belongs to the egg products processing technique field, relate to the preparation method of low irritated albumen.
Background technology
Food hypersenstivity is a kind of bad reaction that body produces some food, has now become the ubiquitous food-safety problem in the whole world.In food hypersenstivity reaction, egg be FAO/WHO assert cause one of 8 large group foods of mankind's allergy, occupy food hypersenstivity ranking list second.According to statistics, there are 0.5%～2.5% children in the whole world to egg allergy, shows as clinically the symptoms such as nettle rash, asthma, stomachache and diarrhoea, is having a strong impact on the healthy of irritated infant and is growing up.
Up to now, egg allergy there is no specific short, and strictly avoiding the edible food containing egg is autopath's optimal selection.But egg often is added in numerous food, avoid edible egg very difficult fully; And concerning the egg degree of allergic reaction low patient, neither optimum method.At present, mainly by processing methods such as Hydrolyze method, heat treatment, glycosylation and fermentations, reduce egg allergen, to reduce the generation of egg allergy.When the Choice and process method reduces the allergenicity of allergenic foods, simultaneously, also should consider organoleptic quality and the nutritive value of food.In these processing methods, hydrolysis is considered to reduce the effective method of allergenic foods sensitization.The fragment that anaphylactogen has an epi-position (linear epitope and comformational epitope) by the protease catalyzing hydrolysis after, its sensitization can significantly reduce; The anaphylactogen hydrolysis also may produce the biologically active peptide of functions such as having immunological regulation, anti-oxidant, hypotensive, and its processing characteristics can improve.But at present, still there are many key issues urgently to be resolved hurrily in the enzyme hydrolysis anaphylactogen: in (1) traditional resolvase protein hydrolysate process, enzyme can produce from being hydrolyzed fragment, disturbs downstream analysis to detect; Simultaneously, hydrolysis is difficult to control, and after hydrolysis finishes, enzyme separates recovery and has certain difficulty, and recycling rate of waterused is low; (2) exposure of a large amount of hydrophobic groupings in hydrolytic process, make hydrolysate with bitter off-flavors.These problems are perplexing and are being hydrolyzed and are affecting its in the extensive use reduced aspect food hypersenstivity originality.
In the last few years, magnetic microsphere is applied to the enzyme immobilization field as carrier becomes study hotspot, enzyme is fixed on magnetic microsphere, both improved the stability of enzyme, also have simultaneously easily separated, reuse continuously and improve the advantages such as immobilized enzyme catalysis efficiency, be conducive to realize serialization and the industrialization of production technology.In view of the problem existed in the harm of egg allergy and current enzyme hydrolysis anaphylactogen technique.The invention provides a kind of method that immobilized enzyme hydrolysis reduces the albumen sensitization, described method comprises: at first use magnetic bead immobilization crosslinking protein enzyme, then utilize immobilization proteinase hydrolysis albumen, hydrolyzate carries out Vacuum Concentration after the de-raw meat of charcoal absorption under 60-70 ℃, and the recycling freeze drying obtains the albumen powder that water content is less than 5%.Low according to the inventive method gained albumen powder sensitization, and contain a large amount of peptide classes and the amino acid of needed by human body, very easily be absorbed by the body, without bad local flavor; There is good promotion and application prospect, can be used as widely autopath's raw-food material.
Summary of the invention
Main purpose of the present invention is to provide a kind of immobilized enzyme hydrolysis to reduce the method for albumen sensitization.Hydrolysis egg-white powder of the present invention nutrition, good palatability, safe, sensitization is low.
For achieving the above object, the technical solution used in the present invention is as follows.
(1) preparation of biotinylation labelled protein enzyme: take a certain amount of biotin, with appropriate 0.02M PBS buffer solution (pH5.5), dissolve, in biotin, with 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimides (EDC), N-hydroxy-succinamide (NHS) mol ratio, be that the 1:3:3 ratio is mixed, reaction 1.5 h under room temperature; Be dissolved in phosphate buffer and prepare protein enzyme solution with any one enzyme in papain, trypsase, Flavourzyme flavor protease, Alcalase alkali protease, the biotin of activation is mixed with protease mol ratio 3:1 ratio, be adjusted to the enzyme optimum pH, after under 4 ℃, reaction is spent the night, remove unreacted biotin with the dialysis of PBS buffer solution, the dialysis product adds glycerine to save backup under-20 ℃.
(2) Streptavidin (SA) coupling magnetic bead: get the magnetic bead of a certain amount of carboxylated, successively with after absolute ethyl alcohol, NaOH, HCl and the washing of PBS buffer solution, with the resuspended magnetic bead of PBS buffer solution.By magnetic bead surfaces carboxyl and EDC, N-hydroxy thiosuccinimide (Sulfo-NHS) mol ratio, be that 1:3:3 adds respectively EDC and Sulfo-NHS, 37 ℃ of oscillating reactions 2 h activation magnetic bead surfaces carboxyls.Magnetic bead reclaims by magnet stand, and, with after PBS buffer solution washing three times, is resuspended in the PBS buffer solution.Magnetic bead after activation adds containing 37 ℃ of reaction 2 h in the PBS buffer solution (pH8.0) of SA, and the final reaction product adds the monoethanolamine room temperature to seal 2 h.Magnet stand reclaims magnetic bead, after the PBS buffer solution washs three times, is resuspended in containing in the PBS buffer solution (pH8.0) of 0.05% Sodium azide, and 4 ℃ save backup.
(3) preparation of immobilised enzymes: immobilised enzymes utilizes Streptavidin MagneSphere and the preparation of biotinylated protein enzyme association reaction, in reaction system, the mol ratio of Streptavidin MagneSphere and biotinylated protein enzyme is 1:1-1:4, and reaction is at 37 ℃ of lower oscillation incubation 20 min.Carry out the magnetic separation through magnetic frame, remove supernatant, reclaim magnetic bead, obtain immobilised enzymes.
(4) albumen solution is mixed by a certain percentage with cushioning liquid, be transferred to afterwards hydrolytic decomposition pot, regulate the temperature of hydrolyzate, the control temperature is 37-60 ℃, pH value 6.0-8.0, then in enzyme, living with the ratio of protein substrate is that 5000-30000 U/g protein adds immobilised enzymes, after mixing, in thermostat water bath, be hydrolyzed, constantly stir during this time, and the pH value 6.0-8.0 of controlled enzymatic hydrolysis system, hydrolysis time is 2.0-4.0 h, and reaction finishes to carry out the magnetic separation by magnetic frame, collects the albumen enzymolysis liquid.
(5) add 0.5-3.0% food stage powdered activated carbon in the albumen enzymolysis liquid that step (4) is collected, hydrolyzate is placed in the water-bath of 50 ℃ and processes 60 min, then through the centrifugal acquisition supernatant of 5000 r/min.
(6) be that 0.08 MPa, temperature are 50 ℃ and carry out Vacuum Concentration by the handled enzymolysis liquid of step (5) in vacuum, be concentrated into solid content more than 60%.
(7) by step (6) gained egg white hydrolyzed peptide concentrate at-40 ℃ of pre-freeze 12 h, condenser temperature is-65 ℃, vacuum 100 Pa, 50 ℃ of temperature of heating plate, vacuum freeze drying 24 h, moisture is below 5%, and dry rear gained powder is low sensitization albumen powder.
Advantage of the present invention is: 1, the present invention can significantly reduce the sensitization of albumen; 2, albumen powder provided by the invention, have absorption easy to digest, is of high nutritive value, and, without bitter off-flavors and other bad flavor, has good promotion and application prospect.
The specific embodiment
The present invention will be described further by following instantiation.
Embodiment 1.
(1) preparation of biotinylation labelled protein enzyme: taking a certain amount of biotin, dissolve with appropriate 0.02M PBS, is that the 1:3:3 ratio is mixed in biotin and EDC, NHS mol ratio, reaction 1.5 h under room temperature; Be dissolved in phosphate buffer and prepare protein enzyme solution with papain, the biotin of activation is mixed with protease mol ratio 3:1 ratio, adjust pH value to 6.0, after reaction is spent the night under 4 ℃, with the dialysis of PBS buffer solution, remove unreacted biotin.
(2) Streptavidin (SA) coupling magnetic bead: get the magnetic bead of a certain amount of carboxylated, after using successively absolute ethyl alcohol, NaOH, HCl and PBS solution washing, with the resuspended magnetic bead of PBS.By magnetic bead surfaces carboxyl and EDC, Sulfo-NHS mol ratio, be that 1:3:3 adds respectively EDC and Sulfo-NHS, 37 ℃ of oscillating reactions 2 h activation magnetic bead surfaces carboxyls.Magnetic bead reclaims by magnet stand, and, with after PBS washing three times, is resuspended in PBS.Magnetic bead after activation adds containing 37 ℃ of reaction 2 h in the PBS buffer solution (pH8.0) of SA, and the final reaction product adds the monoethanolamine room temperature to seal 2 h.Magnet stand reclaims magnetic bead, after PBS washs three times, is resuspended in the PBS(pH8.0 containing 0.05% Sodium azide) in.
(3) preparation of immobilised enzymes: immobilised enzymes utilizes Streptavidin MagneSphere and the preparation of biotinylated protein enzyme association reaction, in reaction system, the mol ratio of Streptavidin MagneSphere and biotinylated protein enzyme is 1:1, and reaction is at 37 ℃ of lower oscillation incubation 20 min.Carry out the magnetic separation through magnetic frame, remove supernatant, reclaim magnetic bead, obtain immobilised enzymes.
(4) albumen solution is mixed by a certain percentage with cushioning liquid, be transferred to afterwards hydrolytic decomposition pot, regulate the temperature of hydrolyzate, controlling temperature is 50 ℃, pH value 6.0, then live with the ratio of protein substrate in enzyme and be that 5000U/g protein adds immobilised enzymes, be hydrolyzed in thermostat water bath after mixing, during constantly stir, and the pH value of controlled enzymatic hydrolysis system, hydrolysis time is 4.0 h, and reaction finishes to carry out the magnetic separation by magnetic frame, collects the albumen enzymolysis liquid.
(5) add 0.5% food stage powdered activated carbon in the albumen enzymolysis liquid that step (4) is collected, hydrolyzate is placed in the water-bath of 50 ℃ and processes 60 min, then through the centrifugal acquisition supernatant of 5000 r/min.
(6) be that 0.08 MPa, temperature are 50 ℃ and carry out Vacuum Concentration by the handled enzymolysis liquid of step (5) in vacuum, be concentrated into solid content more than 60%.
(7) by step (6) gained egg white hydrolyzed peptide concentrate at-40 ℃ of pre-freeze 12 h, condenser temperature is-65 ℃, vacuum 100 Pa, 50 ℃ of temperature of heating plate, vacuum freeze drying 24 h, moisture is below 5%, and dry rear gained powder is low sensitization albumen powder.
Embodiment 2.
(1) preparation of biotinylation labelled protein enzyme: take a certain amount of biotin, with appropriate 0.02M PBS(pH5.5) dissolving, is that the 1:3:3 ratio is mixed in biotin and EDC, NHS mol ratio, reaction 1.5 h under room temperature; Be dissolved in phosphate buffer and prepare protein enzyme solution with trypsase, the biotin of activation is mixed with protease mol ratio 3:1 ratio, adjust pH value to 8.0, after reaction is spent the night under 4 ℃, with the dialysis of PBS buffer solution, remove unreacted biotin.
(2) Streptavidin (SA) coupling magnetic bead: get the magnetic bead of a certain amount of carboxylated, after using successively absolute ethyl alcohol, NaOH, HCl and PBS solution washing, with the resuspended magnetic bead of PBS.By magnetic bead surfaces carboxyl and EDC, Sulfo-NHS mol ratio, be that 1:3:3 adds respectively EDC and Sulfo-NHS, 37 ℃ of oscillating reactions 2 h activation magnetic bead surfaces carboxyls.Magnetic bead reclaims by magnet stand, and, with after PBS washing three times, is resuspended in PBS.Magnetic bead after activation adds containing 37 ℃ of reaction 2 h in the PBS buffer solution (pH8.0) of SA, and the final reaction product adds the monoethanolamine room temperature to seal 2 h.Magnet stand reclaims magnetic bead, after PBS washs three times, is resuspended in the PBS(pH8.0 containing 0.05% Sodium azide) in, 4 ℃ save backup.
(3) preparation of immobilised enzymes: immobilised enzymes utilizes Streptavidin MagneSphere and the preparation of biotinylated protein enzyme association reaction, in reaction system, the mol ratio of Streptavidin MagneSphere and biotinylated protein enzyme is 1:2, and reaction is at 37 ℃ of lower oscillation incubation 20 min.Carry out the magnetic separation through magnetic frame, remove supernatant, reclaim magnetic bead, obtain immobilised enzymes.
(4) albumen solution is mixed by a certain percentage with cushioning liquid, be transferred to afterwards hydrolytic decomposition pot, regulate the temperature of hydrolyzate, controlling temperature is 37 ℃, pH value 8.0, then live with the ratio of protein substrate in enzyme and be that 10000 U/g protein add immobilised enzymes, be hydrolyzed in thermostat water bath after mixing, during constantly stir, and the pH value of controlled enzymatic hydrolysis system, hydrolysis time is 4.0 h, and reaction finishes to carry out the magnetic separation by magnetic frame, collects the albumen enzymolysis liquid.
(5) add 3.0% food stage powdered activated carbon in the albumen enzymolysis liquid that step (4) is collected, hydrolyzate is placed in the water-bath of 50 ℃ and processes 60 min, then through the centrifugal acquisition supernatant of 5000 r/min.
(6) be that 0.08 MPa, temperature are 50 ℃ and carry out Vacuum Concentration by the handled enzymolysis liquid of step (5) in vacuum, be concentrated into solid content more than 60%.
(7) by step (6) gained egg white hydrolyzed peptide concentrate at-40 ℃ of pre-freeze 12 h, condenser temperature is-65 ℃, vacuum 100 Pa, 50 ℃ of temperature of heating plate, vacuum freeze drying 24 h, moisture is below 5%, and dry rear gained powder is low sensitization albumen powder.
Embodiment 3.
(1) preparation of biotinylation labelled protein enzyme: take a certain amount of biotin, with appropriate 0.02M PBS(pH5.5) dissolving, is that the 1:3:3 ratio is mixed in biotin and EDC, NHS mol ratio, reaction 1.5 h under room temperature; Be dissolved in phosphate buffer and prepare protein enzyme solution with the Flavourzyme flavor protease, the biotin of activation is mixed with protease mol ratio 3:1 ratio, adjust pH value to 7.5, after reaction is spent the night under 4 ℃, with the dialysis of PBS buffer solution, remove unreacted biotin.
(2) Streptavidin (SA) coupling magnetic bead: get the magnetic bead of a certain amount of carboxylated, after using successively absolute ethyl alcohol, NaOH, HCl and PBS solution washing, with the resuspended magnetic bead of PBS.By magnetic bead surfaces carboxyl and EDC, Sulfo-NHS mol ratio, be that 1:3:3 adds respectively EDC and Sulfo-NHS, 37 ℃ of oscillating reactions 2 h activation magnetic bead surfaces carboxyls.Magnetic bead reclaims by magnet stand, and, with after PBS washing three times, is resuspended in PBS.Magnetic bead after activation adds containing 37 ℃ of reaction 2 h in the PBS buffer solution (pH8.0) of SA, and the final reaction product adds the monoethanolamine room temperature to seal 2 h.Magnet stand reclaims magnetic bead, after PBS washs three times, is resuspended in the PBS(pH8.0 containing 0.05% Sodium azide) in, 4 ℃ save backup.
(3) preparation of immobilised enzymes: immobilised enzymes utilizes Streptavidin MagneSphere and the preparation of biotinylated protein enzyme association reaction, in reaction system, the mol ratio of Streptavidin MagneSphere and biotinylated protein enzyme is 1:3, and reaction is at 37 ℃ of lower oscillation incubation 20 min.Carry out the magnetic separation through magnetic frame, remove supernatant, reclaim magnetic bead, obtain immobilised enzymes.
(4) albumen solution is mixed by a certain percentage with cushioning liquid, be transferred to afterwards hydrolytic decomposition pot, regulate the temperature of hydrolyzate, controlling temperature is 50 ℃, pH value 7.5, then live with the ratio of protein substrate in enzyme and be that 20000 U/g protein add immobilised enzymes, be hydrolyzed in thermostat water bath after mixing, during constantly stir, and the pH value of controlled enzymatic hydrolysis system, hydrolysis time is 3.0 h, and reaction finishes to carry out the magnetic separation by magnetic frame, collects the albumen enzymolysis liquid.
(5) add 2.0% food stage powdered activated carbon in the albumen enzymolysis liquid that step (4) is collected, hydrolyzate is placed in the water-bath of 50 ℃ and processes 60 min, then through the centrifugal acquisition supernatant of 5000 r/min.
(6) be that 0.08 MPa, temperature are 50 ℃ and carry out Vacuum Concentration by the handled enzymolysis liquid of step (5) in vacuum, be concentrated into solid content more than 60%.
(7) by step (6) gained egg white hydrolyzed peptide concentrate at-40 ℃ of pre-freeze 12 h, condenser temperature is-65 ℃, vacuum 100 Pa, 50 ℃ of temperature of heating plate, vacuum freeze drying 24 h, moisture is below 5%, and dry rear gained powder is low sensitization albumen powder.
Embodiment 4.
(1) preparation of biotinylation labelled protein enzyme: take a certain amount of biotin, with appropriate 0.02M PBS(pH5.5) dissolving, is that the 1:3:3 ratio is mixed in biotin and EDC, NHS mol ratio, reaction 1.5 h under room temperature; Be dissolved in phosphate buffer and prepare protein enzyme solution with the Alcalase alkali protease, the biotin of activation is mixed with protease mol ratio 3:1 ratio, adjust pH value to 8.0, after reaction is spent the night under 4 ℃, with the dialysis of PBS buffer solution, remove unreacted biotin.
(2) Streptavidin (SA) coupling magnetic bead: get the magnetic bead of a certain amount of carboxylated, after using successively absolute ethyl alcohol, NaOH, HCl and PBS solution washing, with the resuspended magnetic bead of PBS.By magnetic bead surfaces carboxyl and EDC, Sulfo-NHS mol ratio, be that 1:3:3 adds respectively EDC and Sulfo-NHS, 37 ℃ of oscillating reactions 2 h activation magnetic bead surfaces carboxyls.Magnetic bead reclaims by magnet stand, and, with after PBS washing three times, is resuspended in PBS.Magnetic bead after activation adds containing 37 ℃ of reaction 2 h in the PBS buffer solution (pH8.0) of SA, and the final reaction product adds the monoethanolamine room temperature to seal 2 h.Magnet stand reclaims magnetic bead, after PBS washs three times, is resuspended in the PBS(pH8.0 containing 0.05% Sodium azide) in.
(3) preparation of immobilised enzymes: immobilised enzymes utilizes Streptavidin MagneSphere and the preparation of biotinylated protein enzyme association reaction, in reaction system, the mol ratio of Streptavidin MagneSphere and biotinylated protein enzyme is 1:4, and reaction is at 37 ℃ of lower oscillation incubation 20 min.Carry out the magnetic separation through magnetic frame, remove supernatant, reclaim magnetic bead, obtain immobilised enzymes.
(4) albumen solution is mixed by a certain percentage with cushioning liquid, be transferred to afterwards hydrolytic decomposition pot, regulate the temperature of hydrolyzate, controlling temperature is 60 ℃, pH value 8.0, then live with the ratio of protein substrate in enzyme and be that 30000 U/g protein add immobilised enzymes, be hydrolyzed in thermostat water bath after mixing, during constantly stir, and the pH value of controlled enzymatic hydrolysis system, hydrolysis time is 2.0 h, and reaction finishes to carry out the magnetic separation by magnetic frame, collects the albumen enzymolysis liquid.
(5) add 1.0% food stage powdered activated carbon in the albumen enzymolysis liquid that step (4) is collected, hydrolyzate is placed in the water-bath of 50 ℃ and processes 60 min, then through the centrifugal acquisition supernatant of 5000 r/min.
(6) be that 0.08 MPa, temperature are 50 ℃ and carry out Vacuum Concentration by the handled enzymolysis liquid of step (5) in vacuum, be concentrated into solid content more than 60%.
(7) by step (6) gained egg white hydrolyzed peptide concentrate at-40 ℃ of pre-freeze 12 h, condenser temperature is-65 ℃, vacuum 100 Pa, 50 ℃ of temperature of heating plate, vacuum freeze drying 24 h, moisture is below 5%, and dry rear gained powder is low sensitization albumen powder.
Claims (2)
1. an immobilized enzyme hydrolysis reduces the method for albumen sensitization, it is characterized in that according to the following steps:
(1) preparation of biotinylation labelled protein enzyme: take a certain amount of biotin, with appropriate pH5.5,0.02M PBS buffer solution, dissolve, in biotin, with 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimides, N-hydroxy-succinamide mol ratio, be that the 1:3:3 ratio is mixed, reaction 1.5 h under room temperature; Be dissolved in phosphate buffer and prepare protein enzyme solution with any one enzyme in papain, trypsase, Flavourzyme flavor protease, Alcalase alkali protease, the biotin of activation is mixed with protease mol ratio 3:1 ratio, be adjusted to the enzyme optimum pH, after under 4 ℃, reaction is spent the night, remove unreacted biotin with the dialysis of PBS buffer solution, the dialysis product adds glycerine to save backup under-20 ℃;
(2) Streptavidin coupling magnetic bead: the magnetic bead of getting a certain amount of carboxylated, successively with after absolute ethyl alcohol, NaOH, HCl and the washing of PBS buffer solution, with the resuspended magnetic bead of PBS buffer solution, by magnetic bead surfaces carboxyl and 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimides, N-hydroxy thiosuccinimide mol ratio, be that 1:3:3 adds respectively 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimides and N-hydroxy thiosuccinimide, 37 ℃ of oscillating reactions 2 h activation magnetic bead surfaces carboxyls.
2. magnetic bead reclaims by magnet stand, and, with after PBS buffer solution washing three times, is resuspended in the PBS buffer solution; Magnetic bead after activation adds 37 ℃ of reaction 2 h of PBS buffer solution containing Streptavidin pH8.0, and the final reaction product adds the monoethanolamine room temperature to seal 2 h; Magnet stand reclaims magnetic bead, after the PBS buffer solution washs three times, is resuspended in the PBS buffer solution of the pH8.0 that contains 0.05% Sodium azide, and 4 ℃ save backup;
(3) preparation of immobilised enzymes: immobilised enzymes utilizes Streptavidin MagneSphere and the preparation of biotinylated protein enzyme association reaction, in reaction system, the mol ratio of Streptavidin MagneSphere and biotinylated protein enzyme is 1:1-1:4, reaction is at 37 ℃ of lower oscillation incubation 20 min, carry out the magnetic separation through magnetic frame, remove supernatant, reclaim magnetic bead, obtain immobilised enzymes;
(4) albumen solution is mixed by a certain percentage with cushioning liquid, be transferred to afterwards hydrolytic decomposition pot, regulate the temperature of hydrolyzate, the control temperature is 37-60 ℃, pH value 6.0-8.0, then in enzyme, living with the ratio of protein substrate is that 5000-30000 U/g protein adds immobilised enzymes, after mixing, in thermostat water bath, be hydrolyzed, constantly stir during this time, and the pH value 6.0-8.0 of controlled enzymatic hydrolysis system, hydrolysis time is 2.0-4.0 h, and reaction finishes to carry out the magnetic separation by magnetic frame, collects the albumen enzymolysis liquid;
(5) add 0.5-3.0% food stage powdered activated carbon in the albumen enzymolysis liquid that step (4) is collected, hydrolyzate is placed in the water-bath of 50 ℃ and processes 60 min, then through the centrifugal acquisition supernatant of 5000 r/min;
(6) be that 0.08 MPa, temperature are 50 ℃ and carry out Vacuum Concentration by the handled enzymolysis liquid of step (5) in vacuum, be concentrated into solid content more than 60%;
(7) by step (6) gained egg white hydrolyzed peptide concentrate at-40 ℃ of pre-freeze 12 h, condenser temperature is-65 ℃, vacuum 100 Pa, 50 ℃ of temperature of heating plate, vacuum freeze drying 24 h, moisture is below 5%, and dry rear gained powder is low sensitization albumen powder.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310345347.5A CN103444979B (en) | 2013-08-09 | 2013-08-09 | Method for lowering albumen allergenicity by immobilized enzyme hydrolysis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310345347.5A CN103444979B (en) | 2013-08-09 | 2013-08-09 | Method for lowering albumen allergenicity by immobilized enzyme hydrolysis |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103444979A true CN103444979A (en) | 2013-12-18 |
| CN103444979B CN103444979B (en) | 2014-10-29 |
Family
ID=49728160
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310345347.5A Expired - Fee Related CN103444979B (en) | 2013-08-09 | 2013-08-09 | Method for lowering albumen allergenicity by immobilized enzyme hydrolysis |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103444979B (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105018569A (en) * | 2015-07-16 | 2015-11-04 | 北京中科紫鑫科技有限责任公司 | Biotin labeling method of ATP sulfurylase |
| CN106978429A (en) * | 2017-03-16 | 2017-07-25 | 中国人民解放军第四军医大学 | A kind of bovine enterokinase light chain load magnetic bead and its preparation method and application |
| CN110144374A (en) * | 2019-05-30 | 2019-08-20 | 吉林大学 | A kind of egg white peptide and preparation method thereof that can promote union of wounded skin |
| CN110463925A (en) * | 2019-08-22 | 2019-11-19 | 华中农业大学 | A kind of method of immobilization proteinase preparation high foaming egg white solution |
| CN110498830A (en) * | 2018-05-16 | 2019-11-26 | 康码（上海）生物科技有限公司 | A kind of preparation method of the magnetic bead for separation and purification of protein |
| CN114524874A (en) * | 2022-03-04 | 2022-05-24 | 广东医科大学 | Method for reducing ovalbumin allergy by laccase/caffeic acid crosslinking and galactomannan combination |
| CN115486523A (en) * | 2021-06-02 | 2022-12-20 | 烟台大学 | Normal-temperature preservation processing method of nutritional seasoning instant sea cucumbers |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0577162A1 (en) * | 1992-04-29 | 1994-01-05 | Genencor International, Inc. | Immobilized enzyme on a carrier of cross-linked gelatin and active carbon |
| CN1596676A (en) * | 2004-08-30 | 2005-03-23 | 江南大学 | Method of modifying lactoalbumin by enzymatic method and its application |
| CN101724675A (en) * | 2008-10-28 | 2010-06-09 | 陈栋梁 | Process for preparing albumin polypeptide |
| CN102429149A (en) * | 2011-11-17 | 2012-05-02 | 吉林大学 | Salted egg white protein polypeptide containing enteral nutrition and preparation method thereof |
| CN102994489A (en) * | 2012-10-09 | 2013-03-27 | 山西农业大学 | Biotin-avidin system immobilized glucoamylase and preparation method thereof |
-
2013
- 2013-08-09 CN CN201310345347.5A patent/CN103444979B/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0577162A1 (en) * | 1992-04-29 | 1994-01-05 | Genencor International, Inc. | Immobilized enzyme on a carrier of cross-linked gelatin and active carbon |
| CN1596676A (en) * | 2004-08-30 | 2005-03-23 | 江南大学 | Method of modifying lactoalbumin by enzymatic method and its application |
| CN101724675A (en) * | 2008-10-28 | 2010-06-09 | 陈栋梁 | Process for preparing albumin polypeptide |
| CN102429149A (en) * | 2011-11-17 | 2012-05-02 | 吉林大学 | Salted egg white protein polypeptide containing enteral nutrition and preparation method thereof |
| CN102994489A (en) * | 2012-10-09 | 2013-03-27 | 山西农业大学 | Biotin-avidin system immobilized glucoamylase and preparation method thereof |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105018569A (en) * | 2015-07-16 | 2015-11-04 | 北京中科紫鑫科技有限责任公司 | Biotin labeling method of ATP sulfurylase |
| CN105018569B (en) * | 2015-07-16 | 2019-05-31 | 北京中科紫鑫科技有限责任公司 | The biotin labeling method of ATP sulfurylase |
| CN106978429A (en) * | 2017-03-16 | 2017-07-25 | 中国人民解放军第四军医大学 | A kind of bovine enterokinase light chain load magnetic bead and its preparation method and application |
| CN106978429B (en) * | 2017-03-16 | 2019-11-15 | 中国人民解放军第四军医大学 | A kind of bovine enterokinase light chain load magnetic bead and its preparation method and application |
| CN110498830A (en) * | 2018-05-16 | 2019-11-26 | 康码（上海）生物科技有限公司 | A kind of preparation method of the magnetic bead for separation and purification of protein |
| CN110144374A (en) * | 2019-05-30 | 2019-08-20 | 吉林大学 | A kind of egg white peptide and preparation method thereof that can promote union of wounded skin |
| CN110463925A (en) * | 2019-08-22 | 2019-11-19 | 华中农业大学 | A kind of method of immobilization proteinase preparation high foaming egg white solution |
| CN115486523A (en) * | 2021-06-02 | 2022-12-20 | 烟台大学 | Normal-temperature preservation processing method of nutritional seasoning instant sea cucumbers |
| CN114524874A (en) * | 2022-03-04 | 2022-05-24 | 广东医科大学 | Method for reducing ovalbumin allergy by laccase/caffeic acid crosslinking and galactomannan combination |
| CN114524874B (en) * | 2022-03-04 | 2023-08-11 | 广东医科大学 | Laccase/caffeic acid cross-linking and galactomannan combined method for reducing ovalbumin allergy |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103444979B (en) | 2014-10-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103444979B (en) | Method for lowering albumen allergenicity by immobilized enzyme hydrolysis | |
| CN102048022B (en) | Soybean polypeptide without hydrolysis and bitter tastes as well as preparation method and application thereof | |
| CN104140992B (en) | A kind of large-scale preparation method of fish scale Type I collagen protein peptides | |
| WO2017076112A1 (en) | Oyster peptide capable of enhancing sexual function, and preparation method and application thereof | |
| JP4384249B2 (en) | Manufacturing method of bee processed food and drink, and bee processed food and drink | |
| CN101294183A (en) | Method for preparing protein peptide with immobilization proteolytic enzyme protolysate | |
| CN104095243A (en) | Preparation technology for enzymolysis bioactive peptide nanometer selenium | |
| CN102296100A (en) | Preparation method of casein antihypertensive peptides | |
| CN108727485A (en) | A kind of preparation method of food-grade hydrolysis of keratin | |
| CN111685286A (en) | Oyster peptide with blood fat reducing function and preparation method and application thereof | |
| CN108893515B (en) | High F value oligopeptide and preparation method thereof | |
| CN103494214A (en) | Casein phosphopeptide and zinc chelate compound | |
| CN110628859A (en) | Glycosylated oyster peptide and preparation method thereof | |
| CN101791039B (en) | Method for preparing high-F value pig blood oligopeptide | |
| CN103652315B (en) | Method for reducing sensitization of soy isolate protein by employing enzyme compound modification | |
| CN104745650B (en) | For the preparation method for the reference state glutamine for improving nutrients in intestine of young pigs | |
| CN102228125A (en) | Preparation method of algal active peptide | |
| CN102823715A (en) | Method for preparing bioactive peptide through full-bionic digestion model method | |
| CN104531663A (en) | Immobilized enzyme hydrolysis method for preparing high F value oligopeptide by using minced squid meat | |
| CN109880874A (en) | A kind of preparation method of rice active peptide, rice active peptide and its application | |
| CN108642113B (en) | Method for efficiently and stably preparing high-F-value corn oligopeptide | |
| CN104046673B (en) | Industrial manufacturing method and application of low-sensitization casein peptide whole powder containing CPPs | |
| CN105907826B (en) | Clean preparation method of plant polypeptide/protein | |
| CN103074404B (en) | Method of using compound enzyme for hydrolysis of whey protein and preparation of peptone | |
| CN103250871A (en) | Hydrolysis process of rice protein |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20141029 Termination date: 20180809 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |