CN110698689A - Preparation method of ovalbumin glycosylation reaction surfactant - Google Patents
Preparation method of ovalbumin glycosylation reaction surfactant Download PDFInfo
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- CN110698689A CN110698689A CN201911146899.7A CN201911146899A CN110698689A CN 110698689 A CN110698689 A CN 110698689A CN 201911146899 A CN201911146899 A CN 201911146899A CN 110698689 A CN110698689 A CN 110698689A
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Abstract
The invention discloses a preparation method of an ovalbumin glycosylation reaction surfactant, which solves the problems that the existing ovalbumin as an emulsifier has insufficient hydrophilicity and needs to be improved in emulsifying capacity. The technical scheme is as follows: (1) the ratio of ovalbumin: dissolving inulin in water at a mass ratio of 0.25-0.5, stirring to dissolve completely, and freeze drying to obtain mixed powder of egg albumin and inulin; (2) placing the powder in an environment with humidity of 60-80%, and reacting for a period of time at a certain temperature to obtain reaction mixture powder; (3) dissolving the reacted mixture powder in water to a concentration of 0.3-0.5g/ml, dialyzing with 6-8kDa dialysis bag for 3-5 days, and freeze-drying to obtain a reaction mixture; the invention has simple process and strong operability, greatly improves the hydrophilicity and the emulsifying capacity of the ovalbumin, and improves the storage stability.
Description
Technical Field
The invention relates to an ovalbumin glycosylation reaction surfactant, in particular to a preparation method of a surfactant for improving the hydrophilicity and the emulsifying capacity of ovalbumin.
Background
Surfactant is known as "industrial monosodium glutamate", and refers to a substance which has fixed hydrophilic and lipophilic groups, can be directionally arranged on the surface of a solution, and can obviously reduce the surface tension. It is a large class of organic compounds, and the properties of the compounds are very characteristic, and the compounds are very flexible and wide in application and have great practical value and theoretical significance. The surfactant at one end isNonpolar hydrocarbon chains (hydrocarbyl groups), which have little affinity for water and are often referred to as hydrophobes; the other end being a polar group (e.g. -OH, -COOH, -NH)2、—SO3H, etc.), which have a large affinity for water, are called hydrophilic groups and are collectively called "amphiphilic molecules" (lipophilic and hydrophilic molecules). At present, the surfactant of ovalbumin (OVA for short) is widely applied to the industries of food, cosmetics, paint and the like, the lipophilicity and emulsification characteristic of the surfactant are well known to be the key, and the good lipophilicity and emulsification characteristic ensure that the cost of required raw materials is reduced, the raw materials are stable and are not easy to generate the phenomenon of creaming to cause the deterioration of products on one hand, and the shelf life of the products can be prolonged on the other hand, so that the quality of the products in the processes of transportation and sale can be ensured. Many products using ovalbumin as a surfactant have been used, one is that raw materials are easy to obtain and low in cost, and the foamability can be applied, but the following are also available: (1) the ovalbumin is poor in water solubility due to insufficient hydrophilicity, and raw materials are wasted to a great extent; (2) due to poor emulsification property, a large amount of other substances are required to be added to maintain the stability of the system, so that the shelf life of the product is greatly shortened; (3) due to the fact that large gaps exist between shells formed among protein macromolecules, the shell layer structure is unstable, and the phenomenon of emulsion breaking is easy to occur.
Disclosure of Invention
In order to solve the technical problems, the invention provides the preparation method of the glycosylated product of the ovalbumin, which has simple process, strong operability, great improvement on the hydrophilicity and the emulsifying capacity of the ovalbumin and improved storage stability.
The technical scheme of the invention comprises the following specific steps:
(1) mixing the egg albumin: dissolving inulin in water at a mass ratio of 0.25-0.5, stirring for 3-5 hr at magnetic stirring frequency of 30-40rpm/min until completely dissolved, and vacuum freeze drying to obtain mixed powder of egg albumin and inulin;
(2) placing the mixed powder in an environment with humidity of 60-80%, at 40-60 deg.C, and reacting for 5-7 days;
(3) dissolving the reacted mixture powder in water to concentration of 0.3-0.5g/ml, dialyzing with 6-8kDa dialysis bag for 3-5 days, and freeze drying the water solution to obtain the reaction product.
Aiming at the problems in the background art, the inventor carries out deep research on the prior egg albumin surfactant, the egg albumin and the inulin are compounded to fully carry out chemical combination reaction at a certain temperature and humidity, and because the inulin is very soluble in water and is combined with the egg albumin to improve the hydrophilicity, and in the aspect of emulsification property, after the inulin is added, polysaccharide and protein carry out glycosylation reaction to change the structure and the molecular weight of the protein, the emulsification activity and the emulsification stability are improved, so that the emulsification performance is improved, stable, firm, uniform-size and full-emulsion is formed under the proportional chemical combination of the egg albumin and the inulin, the emulsification property and the storage stability of the egg albumin are improved, and the development potential and the economic benefit of the egg albumin surfactant are improved.
Most preferably the ratio of ovalbumin to inulin is 0.25. Too much can waste the ovalbumin, improve the cost of raw materials, and too little can cause insufficient reaction, thereby reducing the synthesis rate.
And (2) firstly, carrying out magnetic stirring to fully dissolve the ovalbumin and the inulin, so that the albumen and the inulin can be fully contacted, and the synthesis rate of glycosylation products is increased, wherein the magnetic stirring frequency is preferably 30-40rpm/min, and the stirring time is preferably 3-5 h.
The humidity, temperature and time of the reaction in the step (2) are preferably 60-80%, 40-60 ℃ and 5-7 days, when the humidity is too low, the temperature is too high, and the time is too long, the egg albumin and the inulin are agglomerated, the browning degree is deepened, so that the solubility is reduced, otherwise, the reaction of the egg albumin and the inulin is insufficient or even not reacted, and the raw material cost is increased on the other hand.
As another preferred embodiment of the present invention, the environment having a humidity of 60 to 80% may be an environment having a humidity of 60 to 80% and containing a saturated KBr solution.
In the step (3), redundant impurities are removed by a dialysis method, firstly, the dissolving concentration is preferably 0.3-0.5g/ml, and the insufficient dialysis can be caused by too high concentration; the dialysis band size and dialysis time are preferably 6-8kDa for 3-5 days, and excessive pore size will reduce the reaction product, while too small pore size and too short dialysis time will not remove unreacted ovalbumin and inulin.
The invention has simple process and strong operability, greatly improves the hydrophilicity and the emulsifying capacity of the ovalbumin, and improves the storage stability.
Drawings
FIG. 1 is an infrared spectrum of an ovalbumin glycosylated product;
FIG. 2 is the surface hydrophobicity of the glycated ovalbumin product;
FIG. 3 is the particle size of the egg albumin glycosylation product emulsion;
FIG. 4 is ovalbumin glycosylation product emulsification activity;
figure 5 is the ovalbumin glycosylation product emulsion stability.
Detailed Description
The invention is described in more detail below with reference to specific examples, without limiting the scope of the invention. Unless otherwise specified, the experimental methods adopted by the invention are all conventional methods, and experimental equipment, materials, reagents and the like used in the experimental method can be purchased from chemical companies.
Example 1
(1) Mixing the egg albumin: dissolving inulin in water at a mass ratio of 0.25, stirring to dissolve completely, magnetically stirring at 30rpm/min for 3 hr, and vacuum freeze drying to obtain mixed powder of ovalbumin and inulin;
(2) placing the mixed powder in an environment with the humidity of 60%, wherein the temperature is 40 ℃, and the reaction time is 5 days;
(3) dissolving the reacted mixture powder in water to the concentration of 0.3g/ml, using a 6kDa dialysis bag for 3 days, and freeze-drying the aqueous solution to obtain a reaction product;
(4) adding the pomegranate seed oil into the reaction product solution with the concentration of 1.2%, placing the solution under a high-speed shearing instrument for shearing, and homogenizing for many times by high-pressure microjet to obtain emulsion, wherein the concentration of the pomegranate seed oil in the emulsion is 5% w/w, the rotating speed of the high-speed shearing instrument is 20000rpm/min, the shearing time is 1min, and the homogenizing pressure of the high-pressure microjet is controlled to be 11 kPa. Then observing the emulsifying capacity and the storage stability of the emulsion;
as shown in FIGS. 1-5, the secondary structure of the surfactant was changed in this example compared to unreacted MIX, and the IR spectrum showed a new peak, the surface hydrophobicity was reduced from 2256 to 1872, and the emulsifying activity was increased by 10.61m2The emulsion stability is improved by 12.22h, the size of emulsion droplets is uniform, the particle size is intensively distributed to be 0.691 mu m, and a storage object figure shows that no obvious emulsification phenomenon exists within 14 days, so that the glycosylation product prepared by the invention can be proved to have great improvement on hydrophilicity, emulsion property and storage property.
Example 2
(1) Mixing the egg albumin: dissolving inulin in water at a mass ratio of 0.4, stirring to dissolve completely, magnetically stirring at 35rpm/min for 4 hr, and vacuum freeze drying to obtain mixed powder of ovalbumin and inulin;
(2) placing the mixed powder in an environment with the humidity of 70%, wherein the temperature is 50 ℃, and the reaction time is 6 days;
(3) dissolving the reacted mixture powder in water to the concentration of 0.4g/ml, dialyzing with a 7kDa dialysis bag for 4 days, and freeze-drying the aqueous solution to obtain a reaction product;
(4) adding the pomegranate seed oil into the reaction product solution with the concentration of 1.2%, placing the solution under a high-speed shearing instrument for shearing, and homogenizing for many times by high-pressure microjet to obtain emulsion, wherein the concentration of the pomegranate seed oil in the emulsion is 10% w/w, the rotating speed of the high-speed shearing instrument is 23000rpm/min, the shearing time is 2min, and the homogenizing pressure of the high-pressure microjet is controlled to be 12 kPa. Then observing the emulsifying capacity and the storage stability of the emulsion;
compared with unreacted MIX, the secondary structure of the surfactant is changed, a new substance peak appears in the infrared spectrum, the surface hydrophobicity is reduced from 2256 to 1729, and the emulsifying activity is improved by 18.24m2The emulsion stability is improved by 14.19h, the size of emulsion droplets is uniform, the particle size is intensively distributed to be 0.721 mu m, and a storage object graph shows that no obvious emulsion phenomenon exists within 14 days, so that the glycosylation product prepared by the invention can be proved to have larger improvement on hydrophilicity, emulsion property and storage propertyHigh.
Example 3
(1) Mixing the egg albumin: dissolving inulin in water at a mass ratio of 0.5, stirring to dissolve completely, magnetically stirring at 40rpm/min for 5 hr, and vacuum freeze drying to obtain mixed powder of ovalbumin and inulin;
(2) placing the mixed powder in an environment with the humidity of 80%, wherein the temperature is 60 ℃, and the reaction time is 7 days;
(3) dissolving the reacted mixture powder in water to the concentration of 0.5g/ml, dialyzing with an 8kDa dialysis bag for 5 days, and freeze-drying the aqueous solution to obtain a reaction product;
(4) adding the pomegranate seed oil into the reaction product solution with the concentration of 1.2%, placing the solution under a high-speed shearing instrument for shearing, and homogenizing for many times by high-pressure microjet to obtain emulsion, wherein the concentration of the pomegranate seed oil in the emulsion is 15% w/w, the rotating speed of the high-speed shearing instrument is 25000rpm/min, the shearing time is 3min, and the homogenizing pressure of the high-pressure microjet is controlled to be 13 kPa. Then observing the emulsifying capacity and the storage stability of the emulsion;
compared with unreacted MIX, the secondary structure of the surfactant is changed, a new substance peak appears in the infrared spectrum, the surface hydrophobicity is reduced from 2256 to 1643, and the emulsifying activity is improved by 15.10m2The emulsion stability is improved by 14.19h, the size of emulsion droplets is uniform, the particle size is intensively distributed to be 0.789 mu m, and a storage object graph shows that no obvious emulsification phenomenon exists within 14 days, so that the glycosylation product prepared by the invention can be proved to have great improvement on hydrophilicity, emulsion property and storage property.
The secondary structure of the egg albumin glycosylation product is shown in table 1.
TABLE 1
The above description is only for the purpose of creating a preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art can substitute or change the technical solution and the inventive concept of the present invention within the technical scope of the present invention.
Claims (5)
1. A preparation method of an ovalbumin glycosylation reaction surfactant is characterized by comprising the following steps:
(1) mixing the egg albumin: dissolving inulin in water at a mass ratio of 0.25-0.5, stirring for 3-5 hr at magnetic stirring frequency of 30-40rpm/min until completely dissolved, and vacuum freeze drying to obtain mixed powder of egg albumin and inulin;
(2) placing the mixed powder in an environment with humidity of 60-80%, at 40-60 deg.C, and reacting for 5-7 days;
(3) dissolving the reacted mixture powder in water to concentration of 0.3-0.5g/ml, dialyzing with 6-8kDa dialysis bag for 3-5 days, and freeze drying the water solution to obtain the reaction product.
2. The method for preparing an ovalbumin glycosylation surfactant according to claim 1, wherein the ratio of ovalbumin to inulin is 0.25.
3. The method for preparing ovalbumin glycosylation surfactant according to claim 1, wherein the reaction temperature in the step (2) is 50 ℃ and the reaction time is 6 days.
4. The method for preparing ovalbumin glycosylation surfactant according to claim 1, wherein the environment with humidity of 60-80% is an environment with humidity of 60-80% containing saturated KBr solution.
5. The method for preparing ovalbumin glycosylation surfactant according to claim 1, wherein the dialysis band has a size of 7kDa in step (3) and the dialysis time is 3 days.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101664100A (en) * | 2009-08-24 | 2010-03-10 | 天津春发食品配料有限公司 | Modified and solubilized soybean protein isolate |
| CN103304657A (en) * | 2012-03-16 | 2013-09-18 | 东北林业大学 | Glycosylated modifying method for improving antioxidant activity of whey protein |
| CN108936773A (en) * | 2018-07-03 | 2018-12-07 | 湖北工业大学 | Using ovalbumin-inulin as the preparation method of the microcapsules of wall material |
| CN109247425A (en) * | 2018-08-22 | 2019-01-22 | 湖北工业大学 | A kind of preparation method of the ovalbumin lotion with duplex shell structure |
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- 2019-11-21 CN CN201911146899.7A patent/CN110698689A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101664100A (en) * | 2009-08-24 | 2010-03-10 | 天津春发食品配料有限公司 | Modified and solubilized soybean protein isolate |
| CN103304657A (en) * | 2012-03-16 | 2013-09-18 | 东北林业大学 | Glycosylated modifying method for improving antioxidant activity of whey protein |
| CN108936773A (en) * | 2018-07-03 | 2018-12-07 | 湖北工业大学 | Using ovalbumin-inulin as the preparation method of the microcapsules of wall material |
| CN109247425A (en) * | 2018-08-22 | 2019-01-22 | 湖北工业大学 | A kind of preparation method of the ovalbumin lotion with duplex shell structure |
Non-Patent Citations (1)
| Title |
|---|
| HAO JING等: ""Compar ison of Physicochem ical and Antioxida nt Properties of Egg-W hite Proteins and Fructose and Inulin Maillard Reaction Products"", 《FOOD BIOPROCESS TECHNOL》 * |
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Application publication date: 20200117 |