CN110771893A - Method for preparing β -carotene uniform emulsion from whey protein isolate glycosylation reaction product and uniform emulsion - Google Patents
Method for preparing β -carotene uniform emulsion from whey protein isolate glycosylation reaction product and uniform emulsion Download PDFInfo
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- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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- A—HUMAN NECESSITIES
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- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/10—Foods or foodstuffs containing additives; Preparation or treatment thereof containing emulsifiers
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
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- A23L3/3463—Organic compounds; Microorganisms; Enzymes
- A23L3/3481—Organic compounds containing oxygen
- A23L3/3508—Organic compounds containing oxygen containing carboxyl groups
- A23L3/3517—Carboxylic acid esters
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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Abstract
The invention discloses a method for preparing β -carotene uniform emulsion from whey protein isolate glycosylation reaction products and the uniform emulsion, wherein the method comprises the following steps of 1) preparing the whey protein isolate glycosylation reaction products, 2) preparing a glycosylation protein-polyphenol compound, and 3) preparing β -carotene uniform emulsion.
Description
Technical Field
The invention belongs to the technical field of food science and engineering, and particularly relates to a method for preparing β -carotene uniform emulsion from a glycosylation reaction product of whey protein isolate and the uniform emulsion.
Background
The technical scheme of the invention belongs to the technical field of food science and engineering, and particularly relates to a method for preparing β -carotene uniform emulsion from a glycosylation reaction product of whey protein isolate.
β -carotene is a naturally occurring highly lipophilic carotenoid synthesized by photosynthesis in plants, algae and cyanobacteria, which is becoming increasingly interesting for its various health-promoting benefits, however, due to its extremely low water solubility, chemical and metabolic instability and poor in vivo biological activity, the use of β -carotene in food products, especially nutraceutical fortified food products, has been widely developed and used to protect β -carotene from isomerization, oxidation and degradation, but conventional protein-polysaccharide emulsion systems are susceptible to processing, emulsion stability is susceptible to the external environment, aggregation of active substances in the emulsion, precipitation of active substances from the emulsion, further reduction in the rate of inhibition of the antioxidant activity, and further reduction in the bioavailability of the active substances in the emulsion, due to their extremely low water solubility, chemical and metabolic instability and poor in vivo biological activity.
Disclosure of Invention
The invention aims to solve the problems, adopts a stable uniform emulsion embedding system for separating a protein-polyphenol nano compound from glycosylated whey, can obviously improve the degradation stability of β -carotene in uniform emulsion, effectively prolongs the degradation time of active substances in the emulsion, improves the tolerance of the emulsion embedding system in a complex environment, and provides a simple method for improving the bioavailability of the active substances.
In order to achieve the above object, a first aspect of the present invention provides a method for preparing β -carotene homogeneous emulsion from a glycosylation reaction product of whey protein isolate, the method comprising the steps of:
1) preparation of whey protein isolate glycosylation reaction product
Mixing the whey protein isolate, D- (+) lactose and water to obtain a mixed solution of the whey protein isolate and the D- (+) lactose, adjusting the pH value of the mixed solution to 6.5-7.5, and freeze-drying to obtain protein-lactose mixed powder; placing the protein-lactose mixed powder in an environment with the humidity of 70-79% and the temperature of 65-70 ℃ for reaction to obtain glycosylated whey separated protein powder;
2) preparation of glycosylated protein-polyphenol complexes
Mixing the glycosylated whey protein isolate powder with water to obtain a glycosylated whey protein isolate solution, adding epigallocatechin gallate powder, adjusting the pH of the solution to 3.0-3.3, and fully mixing until the mixed solution turns to milk white to obtain the glycosylated protein-polyphenol compound;
3) β preparation of Carotene homogeneous emulsion
Dissolving β -carotene in an oil phase, centrifuging to remove insoluble substances to obtain β -carotene solution, and mixing and homogenizing the β -carotene solution and the glycosylated protein-polyphenol compound obtained in the step 2) to obtain the β -carotene uniform emulsion.
According to the present invention, water is typically used as laboratory water conventionally used by those skilled in the art, including but not limited to deionized water.
According to the present invention, the pH of the system can be adjusted using an acid-base adjusting agent conventionally used by those skilled in the art. In the step 1), the pH value of the system can be adjusted to 6.5-7.5 by adopting a sodium hydroxide solution; in the step 2), the pH value of the system can be adjusted to 3.0-3.3 by adopting a hydrochloric acid solution.
Preferably, the stirring is performed by magnetic stirring, and the substance is fully dissolved.
Preferably, in the step 1), the mass ratio of the whey protein isolate to the D- (+) lactose is 1: 1 to 2.
Preferably, in the step 1), the total mass of the whey protein isolate and the D- (+) lactose in the mixed solution of the whey protein isolate and the D- (+) lactose is 5 to 10%.
Preferably, in the step 1), the reaction time is 12 to 24 hours.
Preferably, in the step 2), the concentration of the glycated whey protein isolate in the glycated whey protein isolate solution is 4mg/mL-6 mg/mL.
Preferably, in the step 2), the mass ratio of the glycosylated whey protein isolate powder to the epigallocatechin gallate is (1-2): 1 to 2.
Preferably, in the step 3), the concentration of β -carotene in the β -carotene solution is 0.05% -0.1%.
Preferably, in the step 3), the mass ratio of the β -carotene solution to the glycosylated protein-polyphenol compound is 6-7: 3-4.
Preferably, in the step 3), the rotation speed of the centrifugation is 10000 rpm-12000 rpm, and the time is 10 min-15 min.
Preferably, in the step 3), the rotation speed of homogenizing is 7500 rpm-15000 rpm, and the time is 2 min-3 min.
Preferably, in step 3), the oil phase may be rapeseed oil, peanut oil, soybean oil, and the like, and is preferably medium-chain fatty glyceride (MCT).
A second aspect of the invention provides a homogeneous emulsion prepared by the above method.
According to the invention, the homogeneous emulsion has a milk phase volume fraction of about 90% to 100%.
The invention has the beneficial effects that:
the glycosylated protein-polyphenol nanocomposite effectively inhibits coalescence among oil drops due to good emulsification characteristic and interface property, so that emulsion drops show good uniformity, and the uniformity of the emulsion drops is directly related to storage stability. At the same time, the glycosylation reaction makes the protein form a denser and thicker interface layer outside the oil drop loaded with the active substance, which inhibits the interaction between the active substance and the pro-oxidant (metal ions, free radicals, dissolved oxygen). Furthermore, epigallocatechin gallate, which is a polyhydric phenolic substance, has strong antioxidant capacity (such as radical scavenging capacity and metal ion chelating capacity) and can effectively scavenge or chelate pro-oxidant trying to enter liquid drops, thereby improving the retention rate of active substances.
Additional features and advantages of the invention will be set forth in the detailed description which follows.
Drawings
FIG. 1 shows the particle size distribution of β -carotene emulsion prepared in example 1 of the present invention.
FIG. 2 shows a laser confocal map of β -carotene emulsion prepared according to example 1 of the present invention.
FIG. 3 is a graph showing the storage stability analysis of β -carotene emulsion prepared in example 1 of the present invention.
FIG. 4 shows a graph of salt ion stability analysis of β -carotene emulsion prepared in example 1 of the present invention.
FIG. 5 shows a graph of the thermal stability analysis of β -carotene emulsion prepared in example 1 of the present invention.
FIG. 6 shows a stability analysis chart of β -carotene emulsion prepared in example 1 of the present invention.
Detailed Description
Preferred embodiments of the present invention will be described in more detail below. While the following describes preferred embodiments of the present invention, it should be understood that the present invention may be embodied in various forms and should not be limited by the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.
Example 1
This example provides a method for preparing β -carotene homogeneous emulsion from whey protein isolate glycosylation reaction products, comprising the following steps:
1) preparation of whey protein isolate glycosylation reaction product
Mixing the whey protein isolate, D- (+) lactose and water to obtain a mixed solution of the whey protein isolate and the D- (+) lactose, adjusting the pH value of the mixed solution to 7 by adopting a sodium hydroxide solution, and freeze-drying to obtain protein-lactose mixed powder; placing the protein-lactose mixed powder in an environment with the humidity of 70-79% and the temperature of 68 ℃ for reacting for 18h to obtain glycosylated whey separated protein powder;
wherein the mass ratio of the whey protein isolate to the D- (+) lactose is 1: 1.5, the total mass of the whey protein isolate and the D- (+) lactose is 7.5 percent
2) Preparation of glycosylated protein-polyphenol complexes
Mixing the glycosylated whey protein isolate powder with deionized water to obtain a 5mg/mL glycosylated whey protein isolate solution, adding epigallocatechin gallate powder, adjusting the pH of the solution to 3.0-3.3 by using a hydrochloric acid solution, and fully mixing until the mixed solution turns to milk white to obtain the glycosylated protein-polyphenol compound;
wherein the mass ratio of the glycosylated whey protein isolate powder to the epigallocatechin gallate is 1: 1.
3) β preparation of Carotene homogeneous emulsion
β -carotene is dissolved in medium-chain fatty glyceride, insoluble substances are removed through centrifugation (12000rpm, 10min) to obtain β -carotene solution with the concentration of 0.075%, then β -carotene solution and the glycosylated protein-polyphenol compound obtained in the step 2) are mixed and homogenized (10000rpm, 2min), the mass ratio of β -carotene solution to the glycosylated protein-polyphenol compound is 2: 1, and β -carotene uniform emulsion (the emulsion phase volume fraction is 100%) is obtained.
In the steps, a magnetic stirring mode is adopted for mixing, and the substances are ensured to be fully dissolved.
Fig. 1 shows a particle size distribution diagram of β -carotene emulsion prepared in example 1 of the present invention, and fig. 2 shows a laser confocal diagram of β -carotene emulsion prepared in example 1 of the present invention, it can be seen that the structure and particle size distribution of β -carotene uniform emulsion prepared by separating protein-polyphenol nanocomposite from glycosylated whey are that protein is uniformly adsorbed on the surface of emulsion drop, so as to form perfect uniform emulsion structure, uniform particle size distribution and good stability.
FIG. 3 shows a storage stability analysis chart of β -carotene emulsion prepared in example 1 of the present invention, after β -carotene loading, the initial particle size did not change significantly, the particle size of β -carotene loaded by glycosylated WPI-Lac/EGCG emulsion did not change significantly with the extension of storage time, and after 30 days, the emulsion still maintained 50 μm, and had good storage stability.
FIG. 4 is a graph showing the analysis of salt ion stability of β -carotene emulsion prepared in example 1 of the present invention, wherein A is a macroscopic view of β -carotene emulsion at different salt ion concentrations, B is a microscopic view of β -carotene emulsion at a salt ion concentration of 0mM, C is a microscopic view of β -carotene emulsion at a salt ion concentration of 50mM, and E is a microscopic view of β -carotene emulsion at a salt ion concentration of 200mM, and it can be seen from the graph B that the average particle size of WPI-Lac/EGCG emulsion droplets embedding β -carotene slightly increases, gradually increases from 60 μm initially (measured by machine), and when the salt ion concentration increases to 200mM (shown in FIG. E), the particle size increases by 75 μm (measured by machine), but the particle size distribution is still uniform and has a single peak shape, which indicates that the uniform emulsion stabilized by WPI-Lac/EGCG nanoparticles has good salt stability.
FIG. 5 shows a thermal stability analysis diagram of β -carotene emulsion prepared in example 1 of the present invention, wherein A is a macroscopic schematic diagram of β -carotene emulsion processed at different temperatures, B is a microscopic diagram of β -carotene emulsion processed at room temperature (25 ℃), C is a microscopic diagram of β -carotene emulsion processed at 50 ℃ for 15min, and D is a microscopic diagram of β -carotene emulsion processed at 85 ℃ for 15min, as can be seen from the diagrams B-D, the emulsion stabilized by WPI-Lac/EGCG has good thermal stability, strong interfacial adsorption energy, small emulsion particle size change, and the particle size of β -carotene-loaded emulsion is gradually stabilized at about 50 μm (measured by machine), and the emulsion droplets are spherical and have uniform size distribution and are not affected by temperature.
FIG. 6 shows a stability analysis chart of β -carotene emulsion prepared in example 1 of the present invention, the half-life of β -carotene in WPI-Lac/EGCG emulsion reaches 175h, and even emulsion prepared from glycosylated WPI-Lac/EGCG nano-particles has strong protection effect on β -carotene.
Having described embodiments of the present invention, the foregoing description is intended to be exemplary, not exhaustive, and not limited to the embodiments disclosed. Many modifications and variations will be apparent to those of ordinary skill in the art without departing from the scope and spirit of the described embodiments.
Claims (10)
1. A method for preparing β -carotene uniform emulsion from whey protein isolate glycosylation reaction products, which is characterized by comprising the following steps:
1) preparation of whey protein isolate glycosylation reaction product
Mixing the whey protein isolate, D- (+) lactose and water to obtain a mixed solution of the whey protein isolate and the D- (+) lactose, adjusting the pH value of the mixed solution to 6.5-7.5, and freeze-drying to obtain protein-lactose mixed powder; placing the protein-lactose mixed powder in an environment with the humidity of 70-79% and the temperature of 65-70 ℃ for reaction to obtain glycosylated whey separated protein powder;
2) preparation of glycosylated protein-polyphenol complexes
Mixing the glycosylated whey protein isolate powder with water to obtain a glycosylated whey protein isolate solution, adding epigallocatechin gallate powder, adjusting the pH of the solution to 3.0-3.3, and fully mixing until the mixed solution turns to milk white to obtain the glycosylated protein-polyphenol compound;
3) β preparation of Carotene homogeneous emulsion
Dissolving β -carotene in an oil phase, centrifuging to remove insoluble substances to obtain β -carotene solution, and mixing and homogenizing the β -carotene solution and the glycosylated protein-polyphenol compound obtained in the step 2) to obtain the β -carotene uniform emulsion.
2. The method according to claim 1, wherein in the step 1), the mass ratio of the whey protein isolate to the D- (+) lactose is 1: 1 to 2.
3. The method according to claim 1, wherein in the mixed solution of whey protein isolate and D- (+) lactose in step 1), the total mass of whey protein isolate and D- (+) lactose is 5-10%.
4. The method according to claim 1, wherein the reaction time in step 1) is 12 to 24 hours.
5. The method according to claim 1, wherein in step 2), the concentration of glycated whey protein isolate in the glycated whey protein isolate solution is 4mg/mL to 6 mg/mL.
6. The method according to claim 1, wherein in the step 2), the mass ratio of the glycosylated whey protein isolate powder to the epigallocatechin gallate is 1-2: 1 to 2.
7. The method according to claim 1, wherein in step 3), the concentration of β -carotene in the β -carotene solution is 0.05% -0.1%.
8. The method according to claim 1, wherein in the step 3), the mass ratio of the β -carotene solution to the glycosylated protein-polyphenol complex is 6-7: 3-4.
9. The method according to claim 1, wherein, in step 3),
the centrifugal rotating speed is 10000 rpm-12000 rpm, and the time is 10 min-15 min;
the homogenizing rotating speed is 7500 rpm-15000 rpm, and the time is 2 min-3 min.
10. A homogeneous emulsion prepared by the process of any one of claims 1 to 9.
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