CN110101084B - Algae oil nano pickering emulsion and oil powder stabilized by yolk protein peptide particles as well as preparation method and application thereof - Google Patents
Algae oil nano pickering emulsion and oil powder stabilized by yolk protein peptide particles as well as preparation method and application thereof Download PDFInfo
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- CN110101084B CN110101084B CN201910268911.5A CN201910268911A CN110101084B CN 110101084 B CN110101084 B CN 110101084B CN 201910268911 A CN201910268911 A CN 201910268911A CN 110101084 B CN110101084 B CN 110101084B
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Abstract
The invention discloses a seaweed oil nano pickering emulsion and oil powder stabilized by yolk albumin peptide particles, and a preparation method and application thereof. Dispersing defatted yolk protein powder in water, adding trypsin for enzymolysis; inactivating enzyme in boiling water bath, cooling, centrifuging, dialyzing, and lyophilizing to obtain yolk protein peptide granule powder; dissolving yolk protein peptide powder in water, adding algae oil, mixing and stirring; after pre-homogenizing, adopting high-pressure micro-jet treatment to obtain nanoscale algal oil peclin emulsion stabilized by yolk protein peptide particles; and carrying out spray drying treatment on the nano emulsion to obtain an oil powder product rich in omega-3 unsaturated fatty acid. The method has the advantages of simple and mild process conditions, natural and safe raw materials, and capability of carrying out rapid continuous production; the method realizes the reutilization of the waste yolk albumen powder after the oil extraction of the yolk, prepares the nano pickering emulsion rich in omega-3 unsaturated fatty acid and an oil powder ingredient, and can be applied to functional foods, medicines and cosmetics.
Description
Technical Field
The invention relates to a nano pickering emulsion with a particle size less than 200nm and stabilized by yolk protein peptide particles, in particular to a preparation method of a seaweed oil nano pickering emulsion rich in omega-3 polyunsaturated fatty acid (DHA) and oil powder, which are stabilized by yolk protein peptide particles; belongs to the technical field of food, medicine and cosmetic processing.
Background
As a safe food, the eggs contain higher protein and abundant lipid, are good supply sources of vitamins and mineral substances, and have the characteristics of easy absorption, low price and the like, so the eggs are praised as nutritional food for maintaining life. In recent years, there have been breakthrough advances in deep processing of eggs in many countries around the world. Defatted egg yolk powder obtained by deep processing of eggs such as extraction of medicinal lecithin and egg yolk oil is often subjected to waste treatment because of poor functionality, difficulty in water solubility and utilization. The defatted yolk powder contains rich egg protein and has rich nutritional value, and according to the report of related documents, the yolk protein can increase the number of osteoblasts and promote the generation of bones and teeth; activating the motive power of the skeleton and promoting the elongation of the skeleton; promote the calcium deposition of the skeleton and strengthen the toughness of the skeleton; inhibit the growth of osteoclast and maintain the balance of bone components. Therefore, the defatted egg yolk powder is necessary to be fully utilized, and the enzymolysis of the egg yolk albumen powder to convert the egg yolk albumen powder into the peptide with better functionality and utilization rate is one of effective ways for developing and utilizing the defatted egg yolk powder.
The algae oil is extracted from algae, is rich in plant DHA, has strong capability of relieving inflammation, and is easy to be absorbed by human body. DHA is docosahexaenoic acid, belongs to omega-3 polyunsaturated fatty acid, is a main component of human brain and retina lipid, can promote the development of children brain, inhibit platelet aggregation, reduce blood viscosity, delay thrombosis, improve the microcirculation function of organisms, has obvious effect on preventing and treating coronary heart disease and the embolism of cardiovascular and cerebrovascular vessels, and even can eliminate the formed atheromatous plaque. Meanwhile, the algae oil has obvious anti-inflammatory effect, can effectively reduce blood sugar and blood fat, improve the phenomenon of insulin resistance, improve insulin sensitivity and protect islet beta cells. The algae oil contains rich DHA, but is difficult to be widely applied to food due to the poor water solubility, the high oxidation tendency, the unacceptable flavor and the like. The preparation of algal oil emulsions and oil powders is a good strategy to solve this problem.
The research of nanoemulsions (typically 20-200nm in diameter) has also received much attention in recent years; compared with the traditional emulsion, the nano emulsion has better physical stability of resisting sedimentation and elutriation, can be used for loading, encapsulating and protecting hydrophobic active substances, improves the water solubility and bioavailability, and can also be used for embedding medicaments and producing cosmetics. However, the preparation of the nano emulsion at present needs a large amount of synthetic surfactant (20-30 wt%), which is not good for human health. Pickering emulsion is an emulsion stabilized by solid colloidal particles, and because the solid particles are irreversibly adsorbed on an oil/water interface, the emulsion is not sensitive to coalescence, elutriation and Ostwald ripening, and the emulsion stability can reach months or years. In the last five years, a pickering emulsion stabilized by food-grade colloid particles such as protein particles becomes a research hotspot in the field of food colloid and emulsion, and common particle stabilizers for preparing the food-grade pickering emulsion comprise zein particles, soybean protein particles, whey protein micro-colloid particles, wheat gliadin particles, starch particles and the like. Due to the size limitation of food colloidal particles, for example, the particle size of protein or starch particles is generally 100-300nm, the prepared pickering emulsion is micron-sized emulsion, and the oil drop particle size is about 10-100 μm, so that the nano pickering emulsion with the size less than 200nm cannot be prepared by using food-grade colloidal particles at present. Therefore, the preparation of edible nano pickering emulsion with high stability and high safety by using colloidal particles formed by food-grade raw materials such as protein (peptide) and the like for better delivery and stabilization of functional ingredients has become a hot issue of great concern in the field of functional foods and food emulsions.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides a preparation method of the algae oil nano pickering emulsion and the oil powder which are stable by natural, safe, nutritional and stable yolk protein peptide particles.
The invention also aims to provide the application of the algae oil nano pickering emulsion and the oil powder stabilized by the yolk albumin peptide particles in preparing the emulsion rich in omega-3 unsaturated fatty acid and the powdery oil ingredient.
The invention takes the yolk protein peptide particles formed after enzymolysis as a structural unit, is compounded with the algae oil rich in DHA, and prepares the algae oil nano pickering emulsion stabilized by the yolk protein peptide particles through high-pressure homogenization. The nano-scale oil drops are rich in DHA, and the stability and water dispersibility of the nano-scale oil drops can be improved. The invention discovers for the first time that yolk protein peptide obtained by enzymolysis of degreased yolk powder which is a by-product of yolk processing is in a granulation state, can be used for preparing oil-in-water nano pickering emulsion, and stabilizing functional oil such as seaweed oil rich in omega-3 unsaturated fatty acid, thereby expanding the application range of the degreased yolk powder. The invention can not only obtain the nano-scale Pickering emulsion with good stability, but also obtain the grease powder with good re-solubility and rich DHA after spray drying.
From the viewpoint of food nutrition and health, the natural and safe algae oil nano pickering emulsion with good storage stability is prepared by utilizing the interfacial emulsification functional characteristic of the yolk protein peptide particles and controlling simple process conditions. Meanwhile, the emulsion is subjected to spray drying treatment to prepare grease powder rich in DHA, so that the DHA has higher bioavailability in the algae oil nano pickering emulsion.
Eggs are considered to be a natural perfect food. The egg yolk contains abundant protein, and is hydrolyzed by protease to obtain Egg Yolk Protein Hydrolysate (EYPH) containing a large amount of bioactive substances; EYPH is reported to have biological functions of resisting oxidation, reducing blood pressure, reducing blood fat, combining minerals and the like. After the medicinal lecithin and egg yolk oil are extracted, a large amount of defatted protein yellow powder with high protein content is obtained, and the defatted protein yellow powder has poor functionality, is not easy to dissolve in water and is difficult to utilize, and becomes waste in the deep processing process of eggs. The yolk protein peptide with good water solubility and high biological activity is obtained by carrying out enzymolysis on the degreased yolk powder, and is mixed with the algae oil rich in omega-3 unsaturated fatty acid to prepare the nano pickering emulsion and the oil powder, so that the nano pickering emulsion and the oil powder play a good stabilizing role on the algae oil which is easy to oxidize, and the bad flavor generated by the oxidation rancidity of the algae oil can be effectively reduced.
The method carries out trypsin enzymolysis on the degreased yolk protein powder which is the waste after the oil extraction processing of the yolk to obtain the yolk protein peptide with amphipathy, the amphipathy peptide can be self-assembled to form yolk protein peptide nano particles with the size of 10-50nm (the average particle size is 20 nm), the particles can be adsorbed on an oil-water interface, and food-grade Pickering nano emulsion with the particle size of less than 200nm can be prepared by further combining high-pressure micro-jet treatment. The food-grade pickering emulsions reported at present are micron-sized pickering emulsions with particle sizes of 10-100 μm, and may be limited by the size of colloidal particles such as protein or starch (average particle size is 100-300 nm). The invention firstly utilizes food-grade colloidal particles (yolk protein peptide particles) to prepare the nano-scale Pickering emulsion with the granularity less than 200nm, which obviously expands the application range of the Pickering emulsion in the food field.
The nano emulsion carrying system can improve the stability, solubility and bioavailability of active ingredients, and natural protein, polysaccharide and the like can be used as an emulsifier to construct nano emulsion with higher safety, so that the nano emulsion carrying system is widely concerned in the field of food. Common functional active ingredients in food are generally poor in stability, water solubility and bioavailability, are easily affected by environmental factors and gastrointestinal tract digestion, and face the problems of activity maintenance, stability increase, solubility improvement, dispersibility improvement, bioavailability improvement and the like in application. The common solution is to construct a carrier system, such as microcapsules, hydrogels, emulsions, liposomes, etc., to reduce the influence of external factors. Therefore, the application of the nanotechnology to the food industry can enrich product types, improve sensory properties and processability, provide a direction for production and processing in the food field and new product development, and realize modes such as nano emulsion, nano gel, nano microcapsule, nano liposome, nano crystal, lipid nanoparticle and the like in the aspect of carrying functional factors. Compared with the micron-sized pickering emulsion, the nano-sized pickering emulsion has the following obvious advantages: the nano-scale particle size is small, the dynamic stability is high, and the instability rate is slow; the particle size of the emulsion droplets is smaller than the wavelength of incident light, the scattered light is weaker, and a product with certain optical transmittance can be obtained when the particle size and the oil-water phase are proper; the interface area is increased after emulsification, thereby being beneficial to the function of digestive juice and improving the bioavailability of functional active ingredients. The grease such as the seaweed oil of the omega-3 unsaturated fatty acid is rich in polyunsaturated fatty acid, has the functional property of preventing cardiovascular diseases and other chronic diseases, but has higher unsaturation degree and is easy to be oxidized and rancidity, and the constructed functional grease nanoemulsion can obviously improve the oxidation stability and the bioavailability and improve the absorption rate, the absorption amount and the bioavailability of DHA.
The seaweed oil rich in omega-3 unsaturated fatty acid is a liquid functional oil which is easy to oxidize grease, and the pure liquid seaweed oil has poor storage stability, transportability, reprocessing property and the like due to the grease oxidation phenomenon, so that the application of the seaweed oil in the field of functional food is limited. The invention utilizes the nano pickering emulsion stabilized by the yolk protein peptide particles to effectively improve the problems of the functional oil rich in omega-3 unsaturated fatty acid such as the seaweed oil. Firstly, the egg yolk protein particles are adsorbed on an oil-water interface to form an interface film stabilized by polypeptide particles, the long-term physical stability of the algae oil nano emulsion can be realized through a Pickering stabilization mechanism, and the emulsion has uniform appearance and no layering; meanwhile, the close wrapping of the yolk protein peptide particles on the surface of oil drops can keep the stability of the algae oil nano emulsion in the processes of freeze drying and spray drying, so that the algae oil powder with good powder dispersibility, no oil leakage on the surface and good redissolution property can be prepared. In addition, a great deal of research shows that the yolk protein hydrolysate is a peptide mixture with biological activity such as oxidation resistance, so that colloid particles formed by self-assembly of amphiphilic yolk protein peptide are wrapped on the surface of seaweed oil drops to slow down the oxidation degree of grease to a certain extent, further improve the oxidation stability of the seaweed oil nano-emulsion and finally improve the product quality of the seaweed oil and the nano-emulsion and oil powder thereof.
The purpose of the invention is realized by the following technical scheme:
the preparation method of the algae oil nano pickering emulsion and the oil powder stabilized by the yolk albumin peptide particles comprises the following steps:
1) Preparing yolk protein peptide: dispersing the degreased yolk powder in deionized water, heating in water bath, stirring, adjusting pH, adding trypsin for enzymolysis, inactivating enzyme in boiling water bath, cooling, centrifuging, collecting supernatant, dialyzing, and lyophilizing to obtain yolk protein peptide granule powder;
2) Preparing the nano pickering emulsion: dissolving the yolk protein peptide powder in 10mM phosphate buffer solution (pH 7.0), stirring at room temperature for 1h, fully hydrating overnight at 4 ℃, and adding the algae oil, wherein the mass ratio of the yolk protein peptide to the algae oil is controlled to be 1-1; after pretreatment by a homogenizer, high-pressure microjet treatment is adopted to obtain the algae oil nano pickering emulsion stabilized by yolk albumin peptide particles.
3) Preparing oil powder: carrying out spray drying treatment on the algae oil nano pickering emulsion stabilized by the yolk albumin peptide particles; the oil powder rich in omega-3 unsaturated fatty acid with good dispersibility and stability is prepared.
In order to further achieve the purpose of the invention, preferably, the concentration of the defatted egg yolk powder is 0.01-0.08g/mL, the mass ratio of the defatted egg yolk powder to the trypsin is 80.
Preferably, the pH is adjusted to 6.0-10.0, the water bath temperature is 35-45 ℃, the reaction time is 0.5-3h, and the dialysis time is 10-50h.
Preferably, the speed of centrifugation is 6000-10000rpm, the times of centrifugation are 1-3 times, and the time of centrifugation is 10-40min.
Preferably, the concentration of the egg yolk protein peptide is 1-100mg/mL, the mass ratio of the egg yolk protein peptide to the algae oil is 1-1.
Preferably, the speed of the pre-treatment of the homogenizer is 5000-20000rpm, and the time is 1-4min.
Preferably, the pressure of the high-pressure micro-jet is 70-150MPa, and the frequency of the high-pressure micro-jet is 2-5 times.
The algae oil nano pickering emulsion and the oil powder are stabilized by yolk protein peptide particles, DHA in the algae oil is stabilized by the nano emulsion, the algae oil is light yellow, suspended matters do not appear, the emulsion is fine and uniform, and the particle size range is 150-300nm; the emulsion has particle size of 150-300nm after long-term storage, and no precipitation phenomenon occurs during storage.
The algae oil nano pickering emulsion stabilized by the yolk protein peptide particles is subjected to spray drying to obtain DHA grease with good dispersibility; the inlet temperature of the spray drying is 140-180 ℃, and the outlet temperature is 60-80 ℃.
The algae oil nano pickering emulsion and the oil powder stabilized by the yolk albumin peptide particles are used for preparing emulsion rich in omega-3 unsaturated fatty acid and powdery oil ingredients, and are applied to the fields of omega-3 unsaturated fatty acid reinforced functional foods (including formula foods with special medical purposes), medicaments, cosmetics and the like. The alga oil nano pickering emulsion with the stable granulated yolk protein peptide has small scale, and DHA is wrapped in nano-scale oil drops with the stable yolk protein peptide particles, so that the stability and the water dispersibility of the alga oil are improved. The particle size of the emulsion can be effectively reduced by adopting the micro-jet high-pressure homogenization, and when functional food is industrially produced, functional grease ingredients rich in DHA can be produced by adopting the technical method; meanwhile, the oil powder with good re-solubility is obtained after the nano pickering emulsion is subjected to spray drying, a powder ingredient rich in DHA can be prepared and added into functional food, the content of omega-3 unsaturated fatty acid is increased, and the nano pickering emulsion has good effects on reducing blood sugar in a human body, improving the phenomenon of insulin resistance, improving the symptom of hyperlipidemia and the like.
The principle of the invention is as follows: the defatted yolk powder of the invention is rich in high-quality protein, but has poor functionality, is not easy to dissolve in water and is difficult to utilize. The invention discovers that the defatted yolk powder can form amphiphilic yolk protein peptide after enzymolysis, the amphiphilic peptide can be self-assembled to form peptide nanoparticles with the dimension of 10-50nm, good interface adsorption and interface stability are shown, and the nano-scale Pickering emulsion with the yolk protein peptide particle stability dimension of less than 200nm can be prepared after the high-pressure microjet treatment is combined, and the nano-scale Pickering emulsion can effectively stabilize DHA in the algae oil. This stability is mainly conferred by the particle interface layer formed by multiscale distribution of the vitellin nanoparticles at the oil-water interface, and as a whole the vitellin peptides (peptide particles) stabilize the algal oil emulsion droplets in the pickering mechanism. And (3) carrying out spray drying treatment on the stable nano Pickering emulsion to obtain a grease powder product with good redissolution property and rich DHA.
Compared with the prior art, the invention has the following advantages and beneficial effects:
1. the invention provides novel alga oil nano emulsion and powder which are stable by pure natural yolk protein peptide particles and are rich in omega-3 unsaturated fatty acid, and natural and edible nano pickering emulsion is prepared by ingeniously utilizing amphiphilic yolk protein peptide formed by enzymolysis of degreased yolk protein powder and self-assembled particles thereof.
2. The whole process of the invention comprises enzymolysis, high-pressure emulsification and drying, the preparation process is simple, the raw materials are natural, safe and healthy, and no other synthetic antioxidant or auxiliary additive is needed to be added; the product is a nanoemulsion with good stability, small particle size and rich DHA. After drying, a powder product with uniform particles, light yellow appearance and good redissolution property can be obtained.
3. The method has simple and mild process conditions, does not relate to chemical components such as toxic and harmful reagents, emulsifiers and the like, and is green and safe; the method can be used for rapid and continuous production, can be used for preparing the nano emulsion product applied to functional food or cosmetics by simply controlling the process conditions, and has industrial and large-scale application value.
4. The nano pickering emulsion prepared by the method has small scale and good storage stability, can be used for stabilizing the algae oil rich in omega-3 unsaturated fatty acid, and ensures that DHA has high water dispersibility and availability in the algae oil nano pickering emulsion. The invention stabilizes the algae oil by using the interface stability and the biological activity of the yolk protein peptide particles, reduces the contact with oxygen and improves the oxidation stability of the algae oil.
5. When the functional food is industrially produced, the technical method of the invention can be adopted to produce functional oil ingredients such as nano pickering emulsion and oil powder which are rich in omega-3 unsaturated fatty acid; the nano Pickering emulsion with stable yolk protein peptide particles is subjected to spray drying to obtain oil powder with good re-solubility, and the oil powder can be added into functional food to strengthen the content of omega-3 unsaturated fatty acid. Meanwhile, the invention can expand the application range of the degreased yolk albumen powder which is the by-product of yolk oil extraction processing.
Drawings
FIG. 1 is a particle size distribution diagram of a defatted egg yolk protein powder solution and egg yolk protein peptide nanoparticles prepared in example 1 before and after enzymolysis.
FIG. 2 is a particle size distribution diagram of the vitellin-peptide-stabilized nano pickering emulsion with an enzymolysis time of 60min in example 1.
FIG. 3a is a graph showing the appearance of 0.5wt% peptide-5 wt% oil powder of algal oil emulsion from example 1.
FIG. 3b is a graph showing the appearance of 1wt% peptide-5 wt% oil powder of algal oil emulsion in example 1.
FIG. 3c is a graph showing the appearance of 0.5wt% peptide-5 wt% sunflower oil emulsion meal from example 1.
FIG. 3d is a graph showing the appearance of 0.5wt% peptide-5 wt% sunflower oil emulsion meal in example 1.
FIG. 4 is an appearance diagram of the nano Pickering emulsion of 0.5wt% yolk protein peptide-5 wt% algae oil, 1wt% yolk protein peptide-5 wt% algae oil, 0.5wt% yolk protein peptide-5 wt% sunflower oil, 1wt% yolk protein peptide-5 wt% sunflower oil in example 1.
FIG. 5a is the scanning electron microscope image of the powder product obtained by spray drying 0.5wt% peptide-5 wt% algae oil nano pickering emulsion in example 1.
FIG. 5b is the scanning electron microscope image of the powder product obtained by the spray drying treatment of 1wt% peptide-5 wt% algae oil nano pickering emulsion in example 1.
FIG. 5c is the SEM image of the powder product obtained by spray drying 0.5wt% peptide-5 wt% sunflower seed nano pickering emulsion in example 1.
FIG. 5d is the SEM image of the powder product obtained by spray drying of 1wt% peptide-5 wt% sunflower seed Pickering emulsion in example 1.
FIG. 6 is a graph showing the appearance of the Pickering emulsion oil powder of example 2 at 1wt% peptide (10 min) -5wt% algal oil, 1wt% peptide (30 min) -5wt% algal oil, 1wt% peptide (60 min) -5wt% algal oil, 1wt% peptide (120 min) -5wt% algal oil, 1wt% peptide (180 min) -5wt% algal oil, 1wt% peptide (300 min) -5wt% algal oil.
FIG. 7 is an appearance diagram of the powder product obtained by spray drying the yolk protein peptide stabilized nano Pickering emulsion containing 5wt% of algae oil with 1wt% content and different enzymolysis time in example 2.
FIGS. 8 a-8 f are scanning electron micrographs of powders obtained by spray drying of 1wt% vitellin peptide (10 min) -5wt% algal oil, 1wt% vitellin peptide (30 min) -5wt% algal oil, 1wt% vitellin peptide (60 min) -5wt% algal oil, 1wt% vitellin peptide (120 min) -5wt% algal oil, 1wt% vitellin peptide (180 min) -5wt% algal oil, 1wt% vitellin peptide (300 min) -5wt% algal oil nano Pickerlin emulsion, respectively.
Detailed Description
For better understanding of the present invention, the present invention will be further described with reference to the following drawings and examples, but the present invention is not limited thereto.
In the following examples, the particle size was measured as follows:
the nanoemulsion was diluted by 200 times, and the particle size distribution thereof was measured by a Nano-particle sizer (Nano-ZS & MPT-2 type Nano-particle sizer, malvern, uk), the particle refractive index was 1.473, the particle absorption rate was 0.001, the dispersant was water, the dispersant refractive index was 1.330, and the measurement temperature was 25 ℃, to obtain the average particle size (nm) and the particle size volume distribution map.
Example 1
Preparation of yolk protein peptide: dispersing 60g of degreased yolk powder in 1800mL of deionized water, heating in a water bath at 40 ℃, stirring for 0.5h, adjusting the pH value to 8.0 by using a 1M sodium hydroxide solution, adding 0.6g of trypsin for enzymolysis for 60min, keeping the pH value constant in the enzymolysis process, adding 1M hydrochloric acid to adjust the pH value to 7.0 after the enzymolysis is finished, inactivating the enzyme in the boiling water bath, cooling, centrifuging for 30min at the rotation speed of 8000rpm, taking the supernatant, dialyzing for 48h, and freeze-drying to obtain yolk protein peptide particle powder;
preparing a nano pickering emulsion: the yolk peptide powder was dissolved in 10mM phosphate buffer (pH 7.0), stirred at room temperature for 1h, and fully hydrated overnight at 4 ℃ to give a 2wt% yolk peptide solution. Adding 10mM phosphate buffer (pH 7.0), adding different amounts of algae oil and sunflower seed oil to obtain solutions containing 0.5wt% of vitellin peptide-5 wt% algae oil, 1wt% of vitellin peptide-5 wt% algae oil, 0.5wt% of vitellin peptide-5 wt% sunflower seed oil, and 1wt% of vitellin peptide-5 wt% sunflower seed oil; after pretreatment for 2min at 6000rpm of a homogenizer, carrying out high-pressure micro-jet treatment for 3 times at 100MPa to obtain corresponding nano-emulsion; the nanoemulsion was left standing at room temperature overnight, and then measured.
Preparing oil powder: carrying out spray drying treatment on 0.5wt% of yolk protein peptide-5 wt% of seaweed oil, 1wt% of yolk protein peptide-5 wt% of seaweed oil, 0.5wt% of yolk protein peptide-5 wt% of sunflower seed oil and 1wt% of yolk protein peptide-5 wt% of sunflower seed oil nano Pickering emulsion, wherein the inlet temperature is 150 ℃ and the outlet temperature is 80 ℃; the oil powder was left standing overnight at room temperature, and then measured.
FIG. 1 is an appearance diagram of the yolk protein peptide solution and the prepared yolk protein peptide nanoparticle size distribution diagram obtained by performing enzymolysis without adding trypsin and performing enzymolysis for 60min with trypsin in 10mM phosphate buffer (pH 7.0) of defatted yolk protein powder in example 1. It can be known that the defatted egg yolk protein powder solution which is not added with trypsin for enzymolysis has light yellow precipitate, while the egg yolk protein peptide solution which is obtained by adding trypsin for enzymolysis is transparent and has no light yellow precipitate, which indicates that the defatted egg yolk protein powder before enzymolysis is not easy to dissolve in water. According to the particle size distribution diagram of the yolk albumin peptide nanoparticles, the particle size of the yolk albumin peptide nanoparticles is 7-50nm, and the average particle size is 20nm.
FIG. 2 is a particle size distribution diagram of the nanoemulsion stabilized by the yolk protein peptide particles obtained in example 1 with an enzymolysis time of 60 min. As can be seen from FIG. 2, the average particle size of the emulsion was 169.2nm.
FIGS. 3a-3d are graphs showing the appearance of the powder product of example 1, stabilized nanoemulsion containing seaweed oil in an amount of 5% by weight and sunflower oil containing yolk peptide particles in an amount of 0.5% by weight and 1% by weight, after spray drying. As can be seen from FIGS. 3a-3d, the algae oil nano pickering emulsion powder stabilized by the yolk albumin peptide particles is light yellow; the sunflower seed nano pickering emulsion powder stabilized by the yolk protein peptide particles is milky white.
FIG. 4 is a graph showing the appearance of the stabilized nano Pickering emulsion of example 1 containing 0.5wt% and 1wt% of the yolk protein peptide particles and containing 5wt% of algae oil and sunflower seed oil. As can be seen from FIG. 4, the algae oil nano pickering emulsion stabilized by 0.5wt% of the granulated yolk albumin peptide is light yellow, and the phenomenon of oil precipitation occurs on the surface; the algae oil nano pickering emulsion stabilized by 1wt% of yolk albumin peptide particles is light yellow and uniform, and does not separate out oil; the sunflower seed oil nano Pickering emulsion stabilized by 0.5 percent and 1 percent of yolk albumin peptide particles is milky white and uniform and does not separate out oil.
FIGS. 5a-5d are scanning electron microscope images of powder obtained by spray drying of 0.5wt% yolk protein peptide-5 wt% algae oil, 1wt% yolk protein peptide-5 wt% algae oil, 0.5wt% yolk protein peptide-5 wt% sunflower oil, and 1wt% yolk protein peptide-5 wt% sunflower oil nano Pickering emulsion. From fig. 5a-5d, it can be seen that the powder of 0.5wt% vitellin peptide-5 wt% algal oil has holes in the scanning electron microscope, but the powder of 1wt% vitellin peptide-5 wt% algal oil, 0.5wt% vitellin peptide-5 wt% sunflower oil, and 1wt% vitellin peptide-5 wt% sunflower oil can see spherical particles and no holes, which indicates that the powder obtained by spray drying of 1wt% vitellin peptide-5 wt% algal oil, 0.5wt% vitellin peptide-5 wt% sunflower oil, and 1wt% vitellin peptide-5 wt% sunflower oil nano pickering emulsion can load DHA-rich algal oil well.
Yolk protein peptide particles formed by the enzymolysis of the degreased yolk powder can play a role of stabilizing an interface at an oil-water interface; under the action of homogenization, the oil phase and the water phase are fully mixed to prepare the oil-in-water Pickering emulsion. The micro-jet homogenization is to further effectively reduce the size of oil drops under the action of high-pressure shearing to prepare the Pickering emulsion with a nano scale, so that the algae oil and sunflower oil drops are well covered by the yolk protein peptide particles.
As can be seen from the preparation method of the embodiment 1, the raw materials and the reagents used by the invention are natural, green and safe, and the processing process is simple and convenient to operate, thereby being convenient for rapid and continuous production.
In the embodiment, yolk protein peptide particles formed by carrying out enzymolysis on degreased yolk powder are taken as structural units, and the nano-scale pickering emulsion is prepared by adopting micro-jet homogenization. The nano-emulsion prepared by the embodiment can stabilize DHA in nano-oil drops, so that the contact between DHA and oxygen is well reduced, and the stability of DHA is improved. Can provide certain technical support for the industrial production of functional food, medicine and cosmetic ingredients rich in DHA. The invention can realize the deep utilization of the yolk processing waste product-the defatted yolk albumen powder, and promote the sustainable development of the food industry. The food nanoemulsion serving as a functional active ingredient carrying system can embed fat-soluble substances and remarkably improve the stability and the bioavailability, the yolk protein peptide powder can be used for preparing the nanoemulsion with higher stability after enzymolysis, and the nanoemulsion has wide application prospects in the field of foods, such as research and development of functional foods, improvement of the bioavailability of functional active ingredients, improvement of product quality and the like.
Example 2
Preparing yolk protein peptide: dispersing 50g of defatted yolk powder in 1500mL of deionized water, heating in a water bath at 40 ℃ and stirring for 0.5h, adjusting the pH value to 8.0 by 1M sodium hydroxide, adding 0.5g of trypsin for enzymolysis, keeping the pH value of 8.0 constant in the enzymolysis process, sampling at fixed points at different time (10/30/60/120/180/300 min), adjusting the pH value to 7.0 by 1M hydrochloric acid, inactivating enzyme in a boiling water bath, cooling, centrifuging for 30min at the rotation speed of 8000rpm, taking supernatant for dialysis for 48h, and freeze-drying to obtain yolk protein peptide particle powder at different enzymolysis time (10/30/60/120/180/300 min);
preparing the nano pickering emulsion: yolk protein peptide powder was dissolved in 10mM phosphate buffer (pH 7.0), stirred at room temperature for 1h, and fully hydrated at 4 ℃ overnight to give a 2wt% yolk protein peptide solution. Adding 10mM phosphate buffer (pH 7.0) and algae oil to obtain solutions containing 1wt% vitellin peptide (10 min) -5wt% algae oil, 1wt% vitellin peptide (30 min) -5wt% algae oil, 1wt% vitellin peptide (60 min) -5wt% algae oil, 1wt% vitellin peptide (120 min) -5wt% algae oil, 1wt% vitellin peptide (180 min) -5wt% algae oil, and 1wt% vitellin peptide (300 min) -5wt% algae oil, respectively; pretreating for 2min at 6000rpm of a homogenizer, and treating with high-pressure microjet at 100MPa for 3 times to obtain corresponding nanoemulsion; the nanoemulsion was left standing at room temperature overnight, and then measured.
Preparing oil powder: carrying out spray drying treatment on 1wt% of yolk albumen peptide (10 min) -5wt% of algae oil, 1wt% of yolk albumen peptide (30 min) -5wt% of algae oil, 1wt% of yolk albumen peptide (60 min) -5wt% of algae oil, 1wt% of yolk albumen peptide (120 min) -5wt% of algae oil, 1wt% of yolk albumen peptide (180 min) -5wt% of algae oil, 1wt% of yolk albumen peptide (300 min) -5wt% of algae oil nano Pickering emulsion, wherein the inlet temperature is 160 ℃, and the outlet temperature is 70 ℃; the oil powder was left standing overnight at room temperature, and then measured.
The particle size of the emulsion in this example was measured by a nanometer particle sizer. FIG. 6 is a particle size distribution diagram of the stabilized algae oil nano pickering emulsion of the granulated vitellin peptide with different enzymolysis time. As can be seen from FIG. 6, as the enzymolysis time is prolonged, the volume fraction of the emulsion particle size is shifted to the right, the particle size is increased, and the particle size distribution of the emulsion gradually changes from monomodal distribution to bimodal distribution.
FIG. 7 is a graph showing the appearance of the powder product of example 2, which is obtained by spray drying an emulsion containing 5wt% of algal oil stabilized with yolk peptide particles at a content of 1wt% for various enzymatic hydrolysis times. As can be seen from fig. 7, the stabilized algae oil nano pickering emulsion powder of the granulated vitellin obtained from different enzymolysis time is light yellow;
FIGS. 8 a-8 f are scanning electron micrographs of powders obtained by spray drying of 1wt% vitellin peptide (10 min) -5wt% algal oil, 1wt% vitellin peptide (30 min) -5wt% algal oil, 1wt% vitellin peptide (60 min) -5wt% algal oil, 1wt% vitellin peptide (120 min) -5wt% algal oil, 1wt% vitellin peptide (180 min) -5wt% algal oil, 1wt% vitellin peptide (300 min) -5wt% algal oil nano Pickerlin emulsion, respectively. From fig. 8a to fig. 8f, it can be seen that the powder of the algae oil nanoemulsion with stabilized granulated vitellogenin peptide obtained by different enzymolysis time can see spherical particles without holes, which indicates that the powder obtained by spray drying of the nano pickering emulsion can better stabilize the functional oil rich in DHA.
The algae oil nano pickering emulsion prepared by the invention has good storage stability, and an oil powder product with good dispersibility and stability can be obtained by drying treatment. The yolk protein peptide particles adopted in the invention have biological activities such as oxidation resistance and the like, and can improve the oxidation stability of the algae oil in the nano emulsion.
Example 3
Preparing yolk protein peptide: dispersing 50g of degreased yolk powder in 1700mL of deionized water, heating in a water bath at 37 ℃, stirring for 0.5h, adjusting the pH value to 8.0 by using a 1M sodium hydroxide solution, adding 0.5g of trypsin for enzymolysis for 60min, keeping the pH value constant in the enzymolysis process, adding 1M hydrochloric acid to adjust the pH value to 7.0 after the enzymolysis is finished, inactivating the enzyme in the boiling water bath, cooling, centrifuging for 30min at the rotation speed of 8000rpm, taking the supernatant, dialyzing for 48h, and freeze-drying to obtain yolk protein peptide particle powder;
preparing a nano pickering emulsion: yolk peptide powder was dissolved in 10mM phosphate buffer (pH 7.0), stirred at room temperature for 1h, and thoroughly hydrated overnight at 4 ℃ to give a 4wt% yolk peptide solution. Adding 10mM phosphate buffer (pH7.0) and algae oil to obtain algae oil containing yolk protein peptide 3wt% and 10wt%, respectively; after pretreatment for 2min at 5000rpm of a homogenizer, carrying out high-pressure microjet treatment for 3 times at 100MPa to obtain corresponding nano pickering emulsion;
preparing oil powder: the 2wt% yolk protein peptide 10wt% algae oil nano pickering emulsion is subjected to spray drying treatment, the inlet temperature is 150 ℃, the outlet temperature is 70 ℃, and better oil powder can also be obtained.
The micro-jet high-pressure homogenization adopted by the invention can effectively reduce the particle size of the emulsion, and the technology is widely applied to the fields of food, medicine, cosmetic production and the like. Further spray drying the nano Pickering emulsion can prepare an oil powder product which has good dispersibility and stability and is rich in omega-3 unsaturated fatty acid. The method has the advantages of simple and mild process conditions, natural and safe raw materials, capability of rapid continuous and mass production, capability of preparing oil ingredients such as nano pickering emulsion, oil powder and the like rich in omega-3 unsaturated fatty acid by controlling process conditions, application to functional foods, medicines and cosmetics, and industrial and large-scale application values, such as research and development of functional foods, improvement of bioavailability of functional active ingredients, improvement of product quality and the like.
The present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents and fall within the scope of the present invention.
Claims (7)
1. The preparation method of the algae oil nano pickering emulsion and the oil powder stabilized by the yolk albumin peptide particles is characterized by comprising the following steps:
1) Preparing yolk protein peptide particles: dispersing the degreased yolk protein powder in deionized water, heating in a water bath and stirring, adjusting the pH to 6.0-10.0, adding trypsin for enzymolysis, keeping the pH constant in the enzymolysis process, inactivating enzyme in a boiling water bath after the enzymolysis is finished, cooling, centrifuging, taking supernate for dialysis, and freeze-drying to obtain yolk protein peptide particle powder; the mass fraction of the protein of the defatted egg yolk protein powder is more than 90%; the concentration of the defatted egg yolk albumen powder is 0.01-0.08g/mL, the mass ratio of the defatted egg yolk powder to the trypsin is (80); the water bath temperature is 35-45 ℃, and the reaction time is 0.5-3h;
2) Preparing a nano pickering emulsion: dissolving the yolk protein peptide powder in a phosphate buffer solution, stirring for 1h at room temperature, fully hydrating at 4 ℃ overnight, adding the alga oil, and controlling the mass ratio of the yolk protein peptide to the alga oil to be 1-1; after pretreatment by a homogenizer, carrying out high-pressure microjet treatment to obtain a stabilized algae oil nano pickering emulsion containing yolk albumin peptide; the concentration of the yolk protein peptide particles is 1-100mg/mL, the mass ratio of the yolk protein peptide solution to the algae oil is 1-1; the pressure of the high-pressure micro-jet is 70-150MPa, and the frequency of the high-pressure micro-jet is 2-5 times;
3) Preparing oil powder: carrying out spray drying treatment on the algae oil nano pickering emulsion stabilized by the yolk albumin peptide particles; can prepare a grease powder product rich in omega-3 unsaturated fatty acid with good dispersibility and stability.
2. The method for preparing the algae oil nano pickering emulsion and the oil powder stabilized by the yolk protein peptide particles as claimed in claim 1, wherein in the step 1), the dialysis time is 10-50h; the pH is adjusted to 6.0-10.0 by adding sodium hydroxide solution.
3. The method for preparing the algae oil nano pickering emulsion and oil powder stabilized by yolk protein peptide particles as claimed in claim 1, wherein in the step 1), the speed of centrifugation is 6000-10000rpm, the times of centrifugation are 1-3 times, and the time of centrifugation is 10-40min.
4. The method for preparing the algae oil nano pickering emulsion and oil powder stabilized by yolk protein peptide particles as claimed in claim 1, wherein in the step 2), the pre-treatment speed of the homogenizer is 5000-20000rpm, and the time is 1-4min.
5. The algae oil nano pickering emulsion and the oil powder which are prepared by the preparation method and are stabilized by the yolk albumin peptide particles are characterized in that the algae oil nano pickering emulsion is light yellow, has no suspended matters, is fine and uniform and has the particle size of 150-300nm; the emulsion still has the grain diameter of 150-300nm after long-term storage, and does not have the phenomenon of emulsion separation in the storage process, thereby showing good stability.
6. The method for preparing the nano pickering algae oil emulsion and the oil powder stabilized by the yolk albumin peptide particles as claimed in claim 5, wherein the nano pickering algae oil emulsion stabilized by the yolk albumin peptide particles is subjected to spray drying to obtain an oil powder product with good dispersibility; the inlet temperature of the spray drying is 140-180 ℃, and the outlet temperature is 60-80 ℃.
7. The use of the nano pickering emulsion of algae oil stabilized by yolk albumin peptide particles and oil powder as claimed in claim 5 for preparing emulsion and powdery oil ingredient rich in omega-3 unsaturated fatty acid; the emulsion and the powder oil ingredient rich in omega-3 unsaturated fatty acid are applied to functional foods or cosmetics.
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