WO2021169072A1

WO2021169072A1 - Preparation method for pickering emulsion stabilized by fish protein

Info

Publication number: WO2021169072A1
Application number: PCT/CN2020/092491
Authority: WO
Inventors: 高瑞昌; 石彤; 袁丽; 冷伟军; 刘辉
Original assignee: 江苏大学
Priority date: 2020-02-28
Filing date: 2020-05-27
Publication date: 2021-09-02

Abstract

A preparation method for a Pickering emulsion stabilized by fish protein, comprising the following steps: using fish as a raw material to prepare and obtain a myosin solution; preparing an arginine mother liquor or a lysine mother liquor having a certain concentration; mixing the arginine mother liquor or lysine mother liquor with the myosin solution to obtain an aqueous phase; adding an oil phase to the aqueous phase to obtain a mixed solution; shearing and emulsifying the mixed solution to obtain a coarse emulsion; and performing ultrasonic treatment on the coarse emulsion to obtain a Pickering emulsion stabilized by fish myosin. By combining arginine or lysine with myosin, the method may enable the secondary structure of a myosin molecule to transform from a regular structure to an irregular structure, so as to increase the adsorption area of myosin in an oil-water interface, and improving the stability of a Pickering emulsion.

Description

Preparation method of fish protein stable Pickering emulsion

Technical field

The invention belongs to the technical field of food quality improvement, and specifically relates to a preparation method of a Pickering emulsion with stable fish protein.

Background technique

Dietary research has always been the frontier and hotspot of food science research. A large number of functional bioactive ingredients that are beneficial to the human body have gradually been widely used in various food systems. However, due to the fact that such substances are insoluble in water, easy to decompose and have low bioavailability, etc. The reasons limit its application.

In recent years, the construction of a stable Pickering emulsion delivery system for food proteins to replace traditional methods that are difficult to ensure product safety, such as changing the chemical structure or adding surfactants, has gradually become an important factor in improving the biological potency and stability of hydrophobic nutrients. Pathways, and food protein itself is an essential and important component of human nutrition.

In the published Chinese invention patent application, the food proteins used to stabilize Pickering emulsion are mainly soy protein isolate and wheat gliadin. For example, the existing literature discloses a nanoemulsion that improves the stability of pecan oil. Husk pecan oil is prepared into an emulsion to enhance stability and water solubility; it also uses modified soy protein isolate and polyphenols to enhance the synergistic effect to form a soy protein-polyphenol composite emulsion with better stability; and use wheat Gliadin and chitosan acetic acid were used to prepare gelatinous wheat prolamin emulsion.

However, there is no related report on the use of animal protein to stabilize Pickering emulsion. In addition, soy protein isolate has a strong beany flavor and low nutrient utilization, while freshly prepared wheat gliadin has a low pH value and stronger hydrophilicity, which is prone to oil leakage when used in the preparation of Pickering emulsion. The phenomenon. Therefore, looking for animal protein that is more conducive to the stability of Pickering emulsion is of great significance to the improvement of food quality.

Fish meat is very popular among consumers because of its high protein, low fat, and easy digestion and absorption of the protein contained in it. Myofibrillar protein is the most important protein that constitutes fish. It is composed of myosin (55%-60%), actin (20%-25%) and a small amount of tropomyosin and myogenin. Myosin, which accounts for the largest proportion, is selected as the object. This raw material is not only an ideal material for the preparation of Pickering emulsion, but also the selection of this raw material can enrich the diversity of fish protein products and promote the development of my country's freshwater fish industry.

On the other hand, although myosin has the amphiphilic properties required by Pickering emulsions for solid particles, it has limited adsorption capacity at the oil-water interface just like the soy protein isolate commonly used in traditional emulsions, which makes the freshly prepared Pickering emulsions It is difficult to maintain a long shelf life. Therefore, it is very important to improve the stability of Pickering emulsion. It has been reported in the literature that the quaternary structure of the protein is destroyed by ultrasonic cavitation, the solubility of soy protein is increased, and the stability of the emulsion is finally improved. However, this method has higher requirements on the ultrasonic power, temperature and processing time. Industrial application in the field of food quality improvement technology will bring about greater energy loss. Therefore, finding a greener, energy-saving, and environmentally friendly method to prepare a fish protein-stable Pickering emulsion is of great significance to the development of the food industry.

Summary of the invention

In view of the above problems, the present invention aims to solve one of the problems; the present invention cuts from the perspective of myosin secondary structure, evaluates the influence of myosin structure on the stability of Pickering emulsion, and provides a fish protein stable Pickering emulsion The preparation method.

The technical scheme of the present invention is: adding lysine to the freshly prepared myosin to unfold the myosin molecule, and change the secondary structure, so that the secondary structure is changed from the regular type to the non-regular type, and then the modification is changed. As the solid particles in the emulsion, the modified myosin increases the adsorption area of the myosin molecules as the solid particles at the oil-water interface, thereby improving the stability of the emulsion.

In order to achieve the above objective, the present invention provides a method for preparing a fish protein stable Pickering emulsion.

It includes the following steps:

(1) Preparation of myosin: use fish meat as raw material to prepare a solution of myosin;

(2) Prepare a certain concentration of arginine mother liquor or lysine mother liquor;

(3) Mix the arginine mother liquor or lysine mother liquor prepared in step (2) with the myosin solution obtained in step (1) to obtain an aqueous phase;

(4) Add the oil phase to the water phase obtained in step (3) to obtain a mixed solution;

(5) Shearing and emulsifying the mixed solution obtained in step (4) to obtain a coarse emulsion;

(6) The emulsion obtained in step (5) is subjected to ultrasonic treatment to obtain a Pickering emulsion with stable fish protein.

Preferably, the concentration of the arginine mother liquor in step (2) is 400-600 mM; the concentration of the lysine mother liquor is 200-800 mM.

Preferably, the final concentration of arginine in the water phase in step (3) is 5-100 mM; the final concentration of the myosin solution in the water phase is 8-30 mg/mL; the final concentration of lysine in the water phase The final concentration is 20～80mM; the final concentration of myosin solution in the water phase is 10～25mg/mL.

Preferably, the oil phase in step (4) is any one of soybean oil, corn oil, sunflower oil, walnut oil or olive oil.

Preferably, the volume ratio of the water phase to the oil phase in step (4) is (7-9): (1-3).

Preferably, the shear emulsification conditions in step (5) are: 10000 to 30000 rpm, and the time is 1 to 3 minutes.

Preferably, the ultrasonic power in step (6) is 90W-450W, and the ultrasonic time is 3-8min.

The beneficial effects of the present invention:

(1) The selected raw material is fish meat. Fish meat has the characteristics of high protein, low fat and easy digestion and absorption by the human body. There is no relevant report on the preparation of Pickering emulsion using fish protein as the raw material. The method is simple, safe and environmentally friendly; The selection of raw materials can enrich the diversity of fish protein products and promote the development of my country's freshwater fish industry.

(2) In the product prepared by the present invention, arginine or lysine is selected to be combined with myosin, and arginine or lysine can better unfold the myosin molecule and change the secondary structure of the myosin molecule , Make it change from a regular structure to a non-regular structure. This change will greatly increase the adsorption area of myosin at the oil-water interface, and ultimately improve the stability of Pickering emulsion; at the same time, the effect of arginine or lysine The dosage is very important. Too low can not change the secondary structure of myosin well, and too high will bring adverse consequences to the emulsion system; in addition, the dosage of myosin and oil phase is also very important, too low or If it is too high, Pickering emulsion with good performance cannot be prepared.

(3) The use of small molecular amino acids to modify myosin in the present invention can not only achieve the beneficial effect of improving emulsion stability, but also is more green, environmentally friendly and energy-saving than the method of high-intensity ultrasound to improve emulsion stability, and can promote food The development of industry has brought better economic benefits to the society.

Description of the drawings

In FIG. 1, sample B is the freshly prepared emulsion in Comparative Example 1, and sample G is the freshly prepared emulsion in Example 1.

In FIG. 2, sample C is the effect diagram of the emulsion in Comparative Example 1 placed at room temperature for one month, and sample F is the effect diagram of the emulsion in Example 1 placed at room temperature for one month.

Detailed ways

Comparative example 1:

(1) Preparation of myosin:

Reagent A: 0.1M potassium chloride, 20mM Tris, adjust the pH to 7.5 with hydrochloric acid;

Reagent B: 0.45M potassium chloride, 0.2M magnesium acetate, 1mM EGTA, 20mM Tris, 5mM β-mercaptoethanol, adjust the pH to 6.8 with maleic acid;

Reagent C: 0.5M potassium chloride, 20mM Tris, 5mM β-mercaptoethanol, adjust the pH to 7.5 with hydrochloric acid;

Remove the head, viscera, and skin of fresh silver carp, take the white meat from the back, remove the bone, wash, cut into meat paste, add 10 times the volume of reagent A, and homogenize with a homogenizer at 11000r/min for 3~5min. React at 4℃ for 15min, centrifuge (3000×g, 5min, 4℃), take the precipitate, add 5 times the volume of reagent B, add adenosine-5'-triphosphate disodium salt hydrate (ATP ) To make the final concentration 10mM. After mixing, let it stand for 90min at 4℃, then centrifuge (11000×g, 13min, 4℃), take the supernatant, and add 5 times the volume of 1mM potassium bicarbonate. Place for 20 minutes at 4°C, centrifuge (11000×g, 13min, 4°C), take the precipitate, add 2.5 times volume of reagent C, react at 4°C for 10 minutes, add 5 times volume of 1mM potassium bicarbonate, and then add to the mixture Add magnesium chloride to the mixture to make the final concentration of the mixture 10mM, and place it overnight (12-18h) at 4℃; centrifuge the next day (11000×g, 25min, 4℃) to obtain myosin particles, which are added to 0.5 M NaCl-20mM Tris-HCl (pH 7.0) buffer, centrifuge (5000×g, 10min, 4℃), take the supernatant and put it in the refrigerator at 4℃ for later use, adjust the concentration of myosin solution to 15mg/mL, And use within 3 days;

(2) Add soybean oil to step (1) so that the volume ratio of the water phase to the oil phase is 9:1;

(3) Shear emulsify the mixed solution obtained in (2), 12000rpm, 2min.

(4) The coarse emulsion obtained in step (3) is subjected to ultrasonic treatment, 180W, 4min.

Comparative example 2:

(1) Preparation of myosin:

Remove the head, viscera, and skin of fresh silver carp, take the white meat from the back, remove the bone, wash, cut into meat paste, add 10 times the volume of reagent A, and homogenize with a homogenizer at 11000r/min for 3~5min. React at 4℃ for 15min, centrifuge (3000×g, 5min, 4℃), take the precipitate, add 5 times the volume of reagent B, add adenosine-5'-triphosphate disodium salt hydrate (ATP ) To make the final concentration 10mM. After mixing, let it stand for 90min at 4℃, then centrifuge (11000×g, 13min, 4℃), take the supernatant, and add 5 times the volume of 1mM potassium bicarbonate. Place for 20 minutes at 4°C, centrifuge (11000×g, 13min, 4°C), take the precipitate, add 2.5 times volume of reagent C, react at 4°C for 10 minutes, add 5 times volume of 1mM potassium bicarbonate, and then add to the mixture Add magnesium chloride to the mixture to make the final concentration of the mixture 10mM, and place it overnight (12-18h) at 4℃; centrifuge the next day (11000×g, 25min, 4℃) to obtain myosin particles, which are added to 0.5 M NaCl-20mM Tris-HCl (pH 7.0) buffer, centrifuge (5000×g, 10min, 4℃), take the supernatant and put it in the refrigerator at 4℃ for later use, adjust the concentration of myosin solution to 10mg/mL, And use within 3 days;

(3) Shear emulsify the mixed solution obtained in (2), 12000rpm, 2min.

Example 1:

(1) Preparation of myosin:

Remove the head, viscera, and skin of fresh silver carp, take the white meat from the back, remove the bone, wash, cut into meat paste, add 10 times the volume of reagent A, and homogenize with a homogenizer at 11000r/min for 3~5min. React at 4℃ for 15min, centrifuge (3000×g, 5min, 4℃), take the precipitate, add 5 times the volume of reagent B, add adenosine-5'-triphosphate disodium salt hydrate (ATP ) To make the final concentration 10mM. After mixing, let it stand at 4℃ for 90min, then centrifuge (11000×g, 13min, 4℃), take the supernatant, and add 5 times the volume of 1mM potassium bicarbonate, Place for 20 minutes at 4°C, centrifuge (11000×g, 13min, 4°C), take the precipitate, add 2.5 times volume of reagent C, react at 4°C for 10 minutes, add 5 times volume of 1mM potassium bicarbonate, and then add to the mixture Add magnesium chloride to the mixture to make the final concentration of the mixture 10mM, and place it overnight (12-18h) at 4℃; centrifuge the next day (11000×g, 25min, 4℃) to obtain myosin particles, which are added to 0.5 M NaCl-20mM Tris-HCl (pH 7.0) buffer, centrifuge (5000×g, 10min, 4℃), take the supernatant and put it in the refrigerator at 4℃ for later use, adjust the concentration of myosin solution to 15mg/mL, And use within 3 days;

(2) Use the buffer solution 0.5M NaCl-20mM Tris-HCl to dissolve the protein in step (1) to prepare a 400mM lysine mother solution;

(3) Mix the lysine solution prepared in step (2) with the myosin obtained in step (1) to obtain an aqueous phase, wherein the final concentration of lysine in the water phase is 40 mM, and the myosin in the water phase The final concentration of the solution is 15mg/mL;

(4) Add soybean oil to the mixed solution obtained in step (3) so that the volume ratio of the water phase to the oil phase is 9:1;

(5) Shear emulsify the mixed solution obtained in (4), 12000rpm, 2min;

(6) The crude emulsion obtained in step (5) is subjected to ultrasonic treatment at 180 W for 4 min to obtain a Pickering emulsion with stable fish protein.

Example 2:

(1) Preparation of myosin:

Remove the head, viscera, and skin of fresh silver carp, take the white meat from the back, remove the bone, wash, cut into meat paste, add 10 times the volume of reagent A, and homogenize with a homogenizer at 11000r/min for 3~5min. React at 4℃ for 15min, centrifuge (3000×g, 5min, 4℃), take the precipitate, add 5 times the volume of reagent B, add adenosine-5'-triphosphate disodium salt hydrate (ATP ) To make the final concentration 10mM. After mixing, let it stand for 90min at 4℃, then centrifuge (11000×g, 13min, 4℃), take the supernatant, and add 5 times the volume of 1mM potassium bicarbonate. Place for 20 minutes at 4°C, centrifuge (11000×g, 13min, 4°C), take the precipitate, add 2.5 times volume of reagent C, react at 4°C for 10 minutes, add 5 times volume of 1mM potassium bicarbonate, and then add to the mixture Add magnesium chloride to the mixture to make the final concentration of the mixture 10mM, and place it overnight (12-18h) at 4℃; centrifuge the next day (11000×g, 25min, 4℃) to obtain myosin particles, which are added to 0.5 M NaCl-20mM Tris-HCl (pH 7.0) buffer, centrifuge (5000×g, 10min, 4℃), take the supernatant and put it in the refrigerator at 4℃ for later use, adjust the concentration of myosin solution to 18mg/mL, And use within 3 days;

(2) Use 0.5M NaCl-20mM Tris-HCl to dissolve the protein in step (1) to prepare a 200mM lysine mother solution;

(3) Mix the lysine solution prepared in step (2) with the myosin obtained in step (1) to obtain an aqueous phase, wherein the final concentration of lysine in the water phase is 5 mM, and the myosin in the water phase The final concentration of the solution is 18mg/mL;

(4) Add corn oil to the mixed solution obtained in step (3) so that the volume ratio of the water phase to the oil phase is 9:1;

(5) Shear emulsify the mixed solution obtained in (4), 20000rpm, 1.5min;

(6) The crude emulsion obtained in step (5) is subjected to ultrasonic treatment, 90W, 8min, to obtain a fish protein stable Pickering emulsion.

Example 3:

(1) Preparation of myosin:

Remove the head, viscera, and skin of fresh silver carp, take the white meat from the back, remove the bone, wash, cut into meat paste, add 10 times the volume of reagent A, and homogenize with a homogenizer at 11000r/min for 3~5min. React at 4℃ for 15min, centrifuge (3000×g, 5min, 4℃), take the precipitate, add 5 times the volume of reagent B, add adenosine-5'-triphosphate disodium salt hydrate (ATP ) To make the final concentration 10mM. After mixing, let it stand for 90min at 4℃, then centrifuge (11000×g, 13min, 4℃), take the supernatant, and add 5 times the volume of 1mM potassium bicarbonate. Place for 20 minutes at 4°C, centrifuge (11000×g, 13min, 4°C), take the precipitate, add 2.5 times volume of reagent C, react at 4°C for 10 minutes, add 5 times volume of 1mM potassium bicarbonate, and then add to the mixture Add magnesium chloride to the mixture to make the final concentration of the mixture 10mM, and place it overnight (12-18h) at 4℃; centrifuge the next day (11000×g, 25min, 4℃) to obtain myosin particles, which are added to 0.5 M NaCl-20mM Tris-HCl (pH 7.0) buffer, centrifuge (5000×g, 10min, 4℃), take the supernatant and put it in the refrigerator at 4℃ for later use, adjust the concentration of myosin solution to 20mg/mL, And use within 3 days;

(2) Use the buffer solution 0.5M NaCl-20mM Tris-HCl to dissolve the protein in step (1) to prepare 800mM lysine mother liquor;

(3) Mix the lysine solution prepared in step (2) with the myosin obtained in step (1) to obtain an aqueous phase, wherein the final concentration of lysine in the water phase is 40 mM, and the myosin in the water phase The final concentration of the solution is 20mg/mL;

(4) Add walnut oil to the mixed solution obtained in step (3) so that the volume ratio of the water phase to the oil phase is 8:2;

(5) Shear emulsify the mixed solution obtained in (4), 12000rpm, 2min;

(6) The crude emulsion obtained in step (5) is subjected to ultrasonic treatment at 360 W for 5 minutes to obtain a stable Pickering emulsion of fish protein.

Example 4:

(1) Preparation of myosin:

Remove the head, internal organs, and skin of the fresh silver carp, take the white meat from the back, remove the bone, wash, cut into meat paste, add 10 times the volume of reagent A, homogenize with a homogenizer at 11000r/min for 3~5min, React at 4℃ for 15min, centrifuge (3000×g, 5min, 4℃), take the precipitate, add 5 times the volume of reagent B, add adenosine-5'-triphosphate disodium salt hydrate (ATP ) To make the final concentration 10mM. After mixing, let it stand at 4℃ for 90min, then centrifuge (11000×g, 13min, 4℃), take the supernatant, and add 5 times the volume of 1mM potassium bicarbonate. Place for 20 minutes at 4°C, centrifuge (11000×g, 13min, 4°C), take the precipitate, add 2.5 times volume of reagent C, react at 4°C for 10 minutes, add 5 times volume of 1mM potassium bicarbonate, and then add to the mixture Add magnesium chloride to the mixture to make the final concentration of the mixture 10mM, and place it overnight (12-18h) at 4°C; centrifuge the next day (11000×g, 25min, 4°C) to obtain myosin particles, which are added to 0.5 M NaCl-20mM Tris-HCl (pH 7.0) buffer, centrifuge (5000×g, 10min, 4℃), take the supernatant and put it in the refrigerator at 4℃ for later use, adjust the concentration of myosin solution to 15mg/mL, And use within 3 days;

(3) Mix the lysine solution prepared in step (2) with the myosin obtained in step (1) to obtain an aqueous phase, wherein the final concentration of lysine in the water phase is 80 mM, and the myosin in the water phase The final concentration of the solution is 20mg/mL;

(4) Add olive oil to the mixed solution obtained in step (3) so that the volume ratio of the water phase to the oil phase is 7:3;

(5) Perform shear emulsification of the mixed solution obtained in (4), 30000rpm, 1min;

(6) The crude emulsion obtained in step (5) is subjected to ultrasonic treatment at 270 W for 6 min to obtain a Pickering emulsion with stable fish protein.

Example 5:

(1) Preparation of myosin:

Remove the head, viscera, and skin of fresh silver carp, take the white meat from the back, remove the bone, wash, cut into meat paste, add 10 times the volume of reagent A, and homogenize with a homogenizer at 11000r/min for 3~5min. React at 4℃ for 15min, centrifuge (3000×g, 5min, 4℃), take the precipitate, add 5 times the volume of reagent B, add adenosine-5'-triphosphate disodium salt hydrate (ATP ) To make the final concentration 10mM. After mixing, let it stand for 90min at 4℃, then centrifuge (11000×g, 13min, 4℃), take the supernatant, and add 5 times the volume of 1mM potassium bicarbonate. Place for 20 minutes at 4°C, centrifuge (11000×g, 13min, 4°C), take the precipitate, add 2.5 times volume of reagent C, react at 4°C for 10 minutes, add 5 times volume of 1mM potassium bicarbonate, and then add to the mixture Add magnesium chloride to the mixture to make the final concentration of the mixture 10mM, and place it overnight (12-18h) at 4℃; centrifuge the next day (11000×g, 25min, 4℃) to obtain myosin particles, which are added to 0.5 M NaCl-20mM Tris-HCl (pH 7.0) buffer, centrifuge (5000×g, 10min, 4℃), take the supernatant and put it in the refrigerator at 4℃ for later use, adjust the concentration of myosin solution to 30mg/mL, And use within 3 days;

(2) Use 0.5M NaCl-20mM Tris-HCl to dissolve protein in step (1) to prepare 300mM lysine mother liquor;

(3) Mix the lysine solution prepared in step (2) with the myosin obtained in step (1) to obtain an aqueous phase, wherein the final concentration of lysine in the water phase is 60 mM, and the myosin in the water phase The final concentration of the solution is 25mg/mL;

(4) Add peanut oil to the mixed solution obtained in step (3) so that the volume ratio of the water phase to the oil phase is 8:2;

(5) Shear emulsify the mixed solution obtained in (4), 12000rpm, 2min;

(6) The crude emulsion obtained in step (5) is subjected to ultrasonic treatment, and the ultrasonic conditions are 450 W for 3 min, to obtain a Pickering emulsion with stable fish protein.

Example 6:

(1) Preparation of myosin:

(2) Use the buffer solution 0.5M NaCl-20mM Tris-HCl to dissolve the protein in step (1) to prepare a mother liquor with a concentration of 400mM arginine;

(3) Mix the arginine solution prepared in step (2) with the myosin obtained in step (1) to obtain an aqueous phase, wherein the final concentration of arginine in the aqueous phase is 5 mM, and the myosin in the aqueous phase The final concentration of the solution is 8 mg/mL.

(5) Shear emulsify the mixed solution obtained in (4), 12000rpm, 2min;

Example 7:

(1) Preparation of myosin:

(2) Use 0.5M NaCl-20mM Tris-HCl to dissolve the protein in step (1) to prepare a mother liquor with a concentration of 500mM arginine;

(3) Mix the arginine solution prepared in step (2) with the myosin obtained in step (1) to obtain an aqueous phase, wherein the final concentration of arginine in the aqueous phase is 40 mM, and the myosin in the aqueous phase The final concentration of the solution is 15mg/mL;

(5) Shear emulsify the mixed solution obtained in (4), 20000rpm, 1.5min;

(6) The crude emulsion obtained in step (5) is subjected to ultrasonic treatment, and the ultrasonic condition is 270 W for 6 min, to obtain a Pickering emulsion with stable fish protein.

Example 8:

(1) Preparation of myosin:

(2) Use 0.5M NaCl-20mM Tris-HCl to dissolve the protein in step (1) to prepare a 400mM arginine mother liquor;

(3) Mix the arginine solution prepared in step (2) with the myosin obtained in step (1) to obtain an aqueous phase, wherein the final concentration of arginine in the water phase is 100 mM, and the myosin in the water phase The final concentration of the solution is 30mg/mL;

(5) Shear emulsify the mixed solution obtained in (4), 12000rpm, 2min.

(6) The coarse emulsion obtained in step (5) was subjected to ultrasonic treatment, 90W, 8min.

In addition, the B sample in FIG. 1 is the freshly prepared emulsion in Comparative Example 1, and the G sample is the freshly prepared emulsion in Example 1. It can be found that the Pickering emulsion prepared with fish myosin as a raw material can obtain a better quality emulsion, which is milky white and uniformly dispersed.

In FIG. 2, sample C is the effect diagram of the emulsion in Comparative Example 1 placed at room temperature for one month, and sample F is the effect diagram of the emulsion in Example 1 placed at room temperature for one month. It can be found that the C sample is stratified, while the F sample is not stratified, indicating that fish myosin can be used to prepare Pickering emulsion, but the stability of the emulsion decreases after being placed at room temperature for one month, and after mixing with lysine The myosin can more effectively improve the stability of Pickering emulsion.

Table 1 The effect of arginine modification on the content of myosin secondary structure

In Table 1, sample 1 is the myosin described in Comparative Example 2, and sample 2 is the modified myosin described in Example 6. It can be found that the regular structure (α-helix, β-sheet, and β-turn) in fresh myosin (sample 1) is the main part, while after modification with arginine, the irregular structure in myosin (sample 2) The type structure (random curling) is the main body. This transformation is beneficial to increase the adsorption area of myosin molecules at the oil-water interface, thereby improving the stability of the emulsion.

Note: The above embodiments are only used to illustrate the present invention and not to limit the technical solutions described in the present invention; therefore, although this specification has described the present invention in detail with reference to the above-mentioned embodiments, those of ordinary skill in the art should It is understood that the present invention can still be modified or equivalently replaced; and all technical solutions and improvements that do not depart from the spirit and scope of the present invention should be covered by the scope of the claims of the present invention.

Claims

A preparation method of fish protein stable Pickering emulsion, which is characterized in that the specific steps are as follows:

(1) Prepare and obtain myosin solution with fish meat as raw material;

(2) Prepare a certain concentration of arginine mother liquor or lysine mother liquor;

(3) Mix the arginine mother liquor or lysine mother liquor prepared in step (2) with the myosin solution obtained in step (1) to obtain an aqueous phase;

(4) Add the oil phase to the water phase obtained in step (3) to obtain a mixed solution;

(5) Shearing and emulsifying the mixed solution obtained in step (4) to obtain a coarse emulsion;

(6) The emulsion obtained in step (5) is subjected to ultrasonic treatment to obtain a Pickering emulsion with stable fish myosin.
The method for preparing a fish protein stable Pickering emulsion according to claim 1, wherein the concentration of the arginine mother liquor in step (2) is 400-600 mM; the concentration of the lysine mother liquor is 200-600 mM. 800mM.
The method for preparing a fish protein stable Pickering emulsion according to claim 1, wherein the final concentration of arginine in the aqueous phase in step (3) is 5-100 mM; and myosin in the aqueous phase The final concentration of the solution is 8-30mg/mL.
The method for preparing a fish protein stable Pickering emulsion according to claim 1, wherein the final concentration of lysine in the aqueous phase in step (3) is 20-80 mM; The final concentration of the globulin solution is 10-25 mg/mL.
The method for preparing a fish protein-stabilized Pickering emulsion according to claim 1, wherein the oil phase added in step (4) is any of soybean oil, corn oil, sunflower oil, walnut oil or olive oil. A sort of.
The method for preparing a fish protein-stabilized Pickering emulsion according to claim 1, wherein the volume ratio of the water phase and the oil phase in step (4) is (7-9): (1-3) .
The method for preparing a fish protein stable Pickering emulsion according to claim 1, wherein the shear emulsification conditions in step (5) are 10,000 to 30,000 rpm, and the time is 1 to 3 minutes.
The method for preparing a fish protein stable Pickering emulsion according to claim 1, wherein the ultrasonic power in step (6) is 90W-450W, and the ultrasonic time is 3-8min.