CN111067035A

CN111067035A - Egg white peptide pudding containing probiotics and preparation method thereof

Info

Publication number: CN111067035A
Application number: CN202010011059.6A
Authority: CN
Inventors: 姚云昕; 朱冬荣
Original assignee: Zhejiang Aige Biotechnology Co ltd
Current assignee: Zhejiang Aige Biotechnology Co ltd
Priority date: 2020-01-06
Filing date: 2020-01-06
Publication date: 2020-04-28

Abstract

The application discloses a preparation method of egg white peptide pudding containing probiotics, which comprises the following steps: adding protease into the aqueous solution of egg white for enzymolysis to obtain an enzymolysis egg white liquid; adding colloid into the enzymolysis egg white liquid, homogenizing, inoculating lactobacillus, and fermenting at 20-50 deg.C to obtain egg white peptide pudding containing probiotic. And (4) refrigerating the obtained product after fermentation to obtain the product with longer shelf life. The preparation process is improved, a high-temperature method is not used for obtaining a solidification state, and the activity of various components in the pudding is better maintained.

Description

Egg white peptide pudding containing probiotics and preparation method thereof

Technical Field

The application belongs to the technical field of food manufacturing, and particularly relates to egg white peptide pudding containing probiotics and a preparation method thereof.

Background

The eggs are rich in nutrition and are ideal sources of human body protein, amino acid, vitamin and mineral substance. The egg white contains dozens of proteins, mainly including ovalbumin, ovotransferrin, ovomucoid, ovomucin, lysozyme, avidin and the like, and has various active functions.

Bioactive peptides refer to peptide compounds that are beneficial to the vital activities of the living body or have physiological effects, and are also called functional peptides. The bioactive peptide is a general name of different peptides with linear and ring structures formed by various amino acids in different arrangement modes, is a multifunctional complex compound derived from protein, and has unique composition structure and further different active functions. The egg white active peptide is a low molecular weight physiological active peptide formed by acting protease on egg white, the molecular weight of egg white protein, ovoferritin transporter, ovomucoid, lysozyme, ovomucoid avidin and the like in the egg white is reduced after hydrolysis, the biological activities such as oxidation resistance, hypertension resistance, cholesterol resistance and the like are improved, the allergenicity of allergic protein is reduced, the biological active peptide obtained after the egg white is degraded shows good functions of reducing blood pressure, improving immunity and the like, and the egg white active peptide is a good ingredient in the industries such as medicine, food and the like. The biological enzymolysis technology is a degradation process of a substrate by adopting a biological enzyme method under a proper pH value condition, the molecular weight of the polypeptide after enzymolysis is reduced to a certain extent compared with that of protein, and the polypeptide simultaneously shows different physical properties and characteristics and has wide application in the aspects of processing various foods, preparing active products and the like.

Pudding, which is a puzzle sound, is a food solidified from pasty material into solid, such as christmas pudding, bread pudding, yorkshire pudding, etc., and is usually prepared by baking, steaming, baking, etc.; in the narrow sense, pudding refers to a semi-solidified frozen dessert, the main materials of which are egg, cream and the like. The existing preparation method is mainly prepared by a heating method.

Disclosure of Invention

The application provides egg white peptide pudding containing probiotics and a preparation method thereof, and through improvement of a preparation process, a high-temperature method is not used for obtaining a solidified state, so that the activity of various components in the pudding is better maintained.

A preparation method of egg white peptide pudding containing probiotics comprises the following steps:

adding protease into the aqueous solution of egg white for enzymolysis to obtain an enzymolysis egg white liquid; adding colloid into the enzymolysis egg white liquid, homogenizing, inoculating lactobacillus, and fermenting at 20-50 deg.C to obtain egg white peptide pudding containing probiotic bacteria. And (4) refrigerating the obtained product after fermentation to obtain the product with longer shelf life.

According to the preparation method, egg white is prepared into an aqueous solution with a certain concentration, food-grade protease is added for enzymolysis, colloid is added after enzymolysis, then lactobacillus is added for fermentation, and the protein peptide pudding with antioxidant activity is obtained after fermentation.

Several alternatives are provided below, but not as an additional limitation to the above general solution, but merely as a further addition or preference, each alternative being combinable individually for the above general solution or among several alternatives without technical or logical contradictions.

Optionally, the mass percentage of the egg white in the aqueous solution of the egg white is 1% -8%. The water for preparing the egg white aqueous solution is sterile water, and the production environment is 10 ten thousand grade clean environment after the container is sterilized at high temperature or boiled in the production link.

The egg white includes liquid egg white and egg white powder. The liquid egg white is obtained directly from eggs or purchased commercialized liquid egg white; the egg white powder is obtained by extracting egg white from eggs, spray drying or freeze drying, or commercial egg white powder.

Optionally, a step of adding a sweetening agent to the aqueous solution of the egg white is also included before adding the protease. The sweetener is one or more of sucrose, glucose, high fructose syrup, sucralose, stevioside and aspartame. After various sugars are prepared into water solutions with different concentrations, the water solutions are boiled at high temperature and cooled for standby.

Optionally, the method further comprises adjusting the pH value of the aqueous solution of egg white to 2-10.5 before adding the protease. The egg white water solution is adjusted to a certain pH value by adopting food grade hydrochloric acid solution or sodium hydroxide solution, so that the pH value is between 2 and 10.5. Further, before adding the protease, the pH value of the aqueous solution of the egg white is adjusted to 8.5-10.5.

Optionally, the protease is at least one of bromelain, papain, ficin, neutral protease, flavourzyme, alcalase and pepsin.

Optionally, the addition amount of the protease is 0.5-6% of the weight of the aqueous solution of the egg white; the conditions of the enzymolysis treatment are as follows: enzymolysis at 25-60 deg.C for 2-24 hr.

Optionally, the colloid is one or a combination of more of xanthan gum, curdlan, agar, sodium alginate and carrageenan, and the addition proportion is 0.1-0.5% of the mass of the enzymolysis egg white liquid.

Optionally, the homogenization is a homogenization treatment performed after adding the colloid, wherein the homogenization pressure is 10-50MPa, and the homogenization time is 10-50 minutes.

Optionally, the added colloid is a combination of carrageenan and agar, and the added carrageenan: the mass ratio of the two types of agar colloids is 1: at 3, the product has the best elasticity when the total addition is 0.2% (m/m).

Further, after adding the colloid, homogenization treatment was performed at 20MPa for 20 minutes.

Further, the mass percent of egg white in the aqueous solution of the egg white is 5%; adjusting the pH value of the aqueous solution of the egg white to 10.5 before adding the protease; the enzymolysis treatment temperature is 40 ℃; the added colloid is the combination of carrageenan and agar, and the carrageenan: the mass ratio of the two types of agar colloids is 1: 3, the total addition amount is 0.2%; adding colloid, homogenizing at 20MPa for 20 min. The pudding prepared under the condition has the best antioxidant activity and elasticity.

After egg white which is subjected to enzymolysis and is homogenized with colloid is obtained, certain lactic acid bacteria are added for fermentation, and the egg white peptide pudding containing the probiotics with good elasticity and viscosity is obtained, wherein the added lactic acid bacteria are lactic acid bacteria fermentation inoculants allowed to be used in the market.

Optionally, the inoculation amount of the lactic acid bacteria is 0.01-0.5% of the weight of the enzymolysis egg white liquid; the fermentation time is 1-24 h.

The application also provides the egg white peptide pudding containing the probiotics prepared by the preparation method.

Compared with the prior art, the application has at least the following beneficial effects:

(1) the egg white pudding product with self-gelling property is obtained by adopting a proteolysis method, and the protein which is not completely enzymolyzed in the product has activity and is rich in polypeptide active ingredients without high-temperature sterilization.

(2) The concentration and viscoelasticity of the final product can be controlled by adjusting the concentration of the egg white solution and the amount of enzyme added, as well as by adding a small amount of colloidal ingredients.

(3) Lactic acid bacteria are added in the later fermentation process, so that the egg white pudding has better flavor, and the shelf life of the product is prolonged by adding the lactic acid bacteria.

(4) The product retains good active ingredients, has good oxidation resistance, and simultaneously contains a plurality of polypeptides with different molecular weights, so that the product has better and richer nutritive value.

Drawings

FIG. 1 is a process flow diagram of the method of the present application;

FIG. 2 is a graph showing the peptide molecular weight distribution of the sample after pepsin enzymatic hydrolysis in example 1.

FIG. 3 is a graph showing the molecular weight distribution of the resulting polypeptide after hydrolysis by the protease in example 3.

Detailed Description

The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application, and it is obvious that the described embodiments are only a part of the embodiments of the present application, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present application.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein in the description of the present application is for the purpose of describing particular embodiments only and is not intended to be limiting of the application.

Example 1

(1) 5g of sterile egg white powder is dissolved in 75mL of sterile water, and the mixture is stirred and dissolved in a clean sterile container until the mixture is in a uniform state for later use.

(2) Adding 20mL of 50% (m/m) sucrose solution which is heated, boiled and cooled into the solution obtained in the step 1 to obtain the egg white powder solution with the egg white powder concentration of 5% and the sugar solution concentration of 10%.

(3) Adjusting the pH value of egg white aqueous solution to 3.5 by adopting 0.2mol/L food grade hydrochloric acid solution and 0.2mol/L food grade sodium hydroxide solution, then adding 0.5% (m/m) pepsin into the egg white aqueous solution, and carrying out enzymolysis for 12 hours at the temperature of 50 ℃.

(4) After the enzymolysis, 0.2g of Lactobacillus bulgaricus and 0.2g of Streptococcus thermophilus were added to the enzymolyzed egg white, and the mixture was fermented at 42 ℃ for 12 hours.

(5) And after the fermentation is finished, refrigerating the fermented product at 4 ℃ to obtain the egg white peptide pudding with excellent taste.

The process flow of the preparation process is shown in figure 1.

(6) The product is subjected to antioxidant analysis, and the effect of the pudding water extract on eliminating DPPH free radicals and hydroxyl free radicals is analyzed in the experiment. Taking 1g of egg white peptide product, adding 4mL of sterile water, mixing uniformly and centrifuging, and using the supernatant for sample antioxidant activity analysis:

(6.1) measurement of DPPH scavenging ability

mu.L of the sample was added to 4mL of a 0.1mmol/L DPPH-methanol solution, followed by 450. mu.L of a 50mmol/L LTris-HCl buffer (7.4), and incubated at 25 ℃ for 30min in a thermostatic water bath. Deionized water was used as a reference solution. Absorbance was measured at 517 nm.

DPPH radical scavenging ability (%) - (A)₀-(A₁-A₂))/A₀]×100

In the formula: a. the₀The absorbance of the blank control solution; a. the₁For the determination of samplesThe absorbance of the tube; a. the₂Absorbance of the bottom tube is the sample.

(6.2) reduction force measurement

mu.L of the sample was added to 2.5mL of phosphate buffer (0.2mol/L, pH 6.6), 2.5mL of 1% potassium ferricyanide (w/v) was added, the reaction was carried out at 50 ℃ for 30min, 2.5mL of 10% trichloroacetic acid (w/v) was added, the mixture was centrifuged at 3000rpm for 10min, and 2.5mL of the supernatant was immediately taken, 2.5mL of deionized water was added, and 0.5mL of 0.1% ferric trichloride (w/v) was added. Deionized water was used as a reference solution. Measured at 700 nm. The higher the absorbance value, the stronger the reducing power.

(6.3) measurement of hydroxyl radical scavenging ability

150 μ L of sample was added to 1.4mL of 6mmol/L H₂O₂Then, 0.6mL of 20mmol/L sodium salicylate and 2mL of 1.5mmol/L ferrous sulfate were added, and the mixture was subjected to thermostatic water bath at 37 ℃ for 1 hour. Deionized water was used as a reference solution. Absorbance was measured at 562 nm.

Hydroxyl radical scavenging ability (%) - (A)₀-(A₁-A₂))/A₀]×100

In the formula: a. the₀The absorbance of the blank control solution; a. the₁Determining the absorbance of the tube for the sample; a. the₂Absorbance of the bottom tube is the sample.

The results of analysis of DPPH scavenging ability, reducing ability and hydroxyl radical scavenging ability are shown in Table 1.

(6.4) measurement of peptide molecular weight

And (3) determining by adopting laser matrix analysis mass spectrometry: the sample was dissolved in a solvent and applied to a test plate in small amounts, and the test plate was dried naturally and placed in a device for testing, the results of which are shown in fig. 2.

As shown in FIG. 2, after the egg white protein is subjected to enzymolysis, different polypeptides with molecular weight of 500-3000 are generated, wherein the polypeptide with the maximum ratio of 600-1000 and high strength, such as the polypeptide with the molecular weight of 640.3, is higher than the polypeptide with the molecular weight of 2400-2800, which also indicates that the protein in the egg white is degraded into low molecular weight peptide.

TABLE 1 antioxidant Effect of different enzyme preparations on degradation of protein substrates

	DPPH radical scavenging ability	Reducing power	Hydroxyl radical scavenging ability
				Example 1 sample	51.2％	0.399	46.7％

Example 2

(1) Dissolving appropriate amount of egg white powder in water, stirring and dissolving in a clean sterile container to uniform state for use;

(2) determining optimal parameters of the process of preparing the egg white protein peptide by enzymolysis by using the egg white powder dosage, the enzymolysis pH value and the enzymolysis temperature through an orthogonal experiment, wherein the factor level table is shown in table 2; wherein the egg white powder is accurately weighed, the enzymolysis pH value is prepared by 0.2mol/L food-grade hydrochloric acid and sodium hydroxide solution, and the enzymolysis temperature is controlled by a water bath kettle. In this example, alkaline protease was used as the protease, and the amount of the enzyme added was 1% (m/m), and other parameters are shown in Table 2. (3) Adding a proper amount of sterile sucrose solution after enzymolysis, and uniformly mixing to control the mass concentration to be 4% (V/V);

(4) according to the following carrageenin: the ratio of the two types of colloids of agar is 1: 1,1: 2,1: 3,2: 1,2: 3,3: 2, the total addition amount is 0.1 percent and 0.2 percent, and the homogenization treatment is carried out for 20 minutes under 20MPa after the uniform stirring.

(5) To the above solution, 0.2% of Lactobacillus bulgaricus powder and 0.2% of Streptococcus thermophilus powder were added, respectively, and fermented at 43 ℃ for 24 hours.

(6) And (3) after the fermentation is finished, refrigerating the fermented product at 4 ℃ to obtain the pudding product containing the egg white peptide.

(7) Antioxidant analysis is carried out on the product, and the effect of the pudding water extract on eliminating DPPH free radicals and hydroxyl free radicals is demonstrated in the experiment. 10g of egg white peptide product is added with 40mL of sterile water, mixed evenly and centrifuged, and the supernatant is used for the analysis of the antioxidant activity of the sample. The soluble protein content was measured by Coomassie brilliant blue colorimetry, the other methods were the same as in example 1, and the elasticity was measured by texture analyzer method as follows.

Selecting a P/10 type probe by the texture analyzer, testing the speed of the test card by 5mm/s, and testing the speed: 3mm/s, speed after test: 5mm/s, trigger force: 5g, undercut displacement: 5 mm. The analysis results are shown in tables 2 to 5.

TABLE 2 orthogonal experiment factor horizon

Level of factor	pH value	Concentration of egg white powder	Temperature of enzymolysis
				1	8.5	2％	40
2	9.5	5％	50
				3	10.5	10％	60

TABLE 3 orthogonal experimental results Table

TABLE 4 viscosity number of samples

The protein content, viscosity and oxidation performance of the final product can be controlled by adjusting the concentration of the egg white solution, the pH value of the egg white solution and the enzymolysis temperature, and the 8 th group is the group with the best antioxidant activity through measurement and calculation.

The elasticity of the product can be adjusted by adding a proper amount of colloid, and the elasticity values of the product under different colloids and different colloid addition amounts are compared under the condition of the 8 th group, and the results are shown in the table 5:

TABLE 5 sample elasticity values

Example 3

(1) 4000g of sterile egg white powder is dissolved in 76L of sterile water, and the mixture is stirred and dissolved in a clean sterile container until the mixture is uniform for later use.

(2) Adding 4000g of sucrose into 18L of sterile water, heating to boil, cooling, adding into 80L of water obtained in the step 1, and mixing uniformly to obtain the egg white powder solution with the egg white powder concentration of 4% and the sugar solution concentration of 4%.

(3) The method comprises the steps of adopting a 2mol/L food grade hydrochloric acid solution and a 2mol/L food grade sodium hydroxide solution to adjust the pH value of egg white aqueous solution to 9, then adding 0.6% (m/m) alkaline protease into the egg white aqueous solution, and carrying out enzymolysis for 24 hours at the temperature of 50 ℃.

(4) After enzymolysis, 200g of lactobacillus bulgaricus powder and 200g of streptococcus thermophilus powder are added into the enzymolysis egg white, and fermentation is carried out for 24 hours at the temperature of 45 ℃.

(5) And (3) after the fermentation is finished, refrigerating the fermented product at 4 ℃ to obtain the pudding product containing the egg white peptide. The results of the molecular weight measurements are shown in FIG. 3.

As shown in FIG. 3, the molecular weight distribution of the obtained polypeptide is between 2000-5500 after the egg white is subjected to enzymolysis by using alkaline protease, wherein the strength is higher between 3500-5000.

The above-mentioned embodiments only express several embodiments of the present application, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the concept of the present application, which falls within the scope of protection of the present application. Therefore, the protection scope of the present patent shall be subject to the appended claims.

Claims

1. A preparation method of egg white peptide pudding containing probiotics is characterized by comprising the following steps:

adding protease into the aqueous solution of egg white for enzymolysis to obtain an enzymolysis egg white liquid; adding colloid into the enzymolysis egg white liquid, homogenizing, inoculating lactobacillus, and fermenting at 20-50 deg.C to obtain egg white peptide pudding containing probiotic bacteria.

2. The preparation method of claim 1, wherein the mass percentage of egg white in the aqueous solution of egg white is 1-8%.

3. The method of claim 1, further comprising the step of adding a sweetener to the aqueous solution of egg white prior to adding the protease.

4. The method of claim 1, further comprising adjusting the pH of the aqueous solution of egg white to 2-10.5 prior to adding the protease.

5. The method according to claim 1, wherein the protease is at least one of bromelain, papain, ficin, neutral protease, flavourzyme, alcalase and pepsin.

6. The preparation method according to claim 1, wherein the protease is added in an amount of 0.5-6% by weight based on the weight of the aqueous solution of egg white; the conditions of the enzymolysis treatment are as follows: enzymolysis at 25-60 deg.C for 2-24 hr.

7. The preparation method according to claim 1, wherein the colloid is one or a combination of more of xanthan gum, curdlan, agar, sodium alginate and carrageenan, and the addition proportion is 0.1-0.5% of the mass of the enzymolysis egg white liquid.

8. The method according to claim 1, wherein the homogenization is a homogenization treatment after adding the colloid, and the homogenization pressure is 10-50MPa and the homogenization time is 10-50 minutes.

9. The method according to claim 1, wherein the amount of the lactic acid bacteria is 0.01-0.5% by weight of the enzymatically hydrolyzed egg white; the fermentation time is 1-24 h.

10. The egg white peptide pudding containing the probiotics prepared by the preparation method of any one of claims 1 to 9.