AU2019204868A1 - Production method of high bioactivity whole egg fermentation, and products and applications thereof - Google Patents

Abstract

Disclosed is a method of producing high bioactivity whole egg fermentation, which comprises step 1 in which poultry eggs were cleaned, sterilized and aired, into which edible organic acid is then added by 1-3 times the weight of the poultry eggs to submerged and soaked the eggs; step 2 in which the egg was stirred to release the egg membrane and the egg liquid followed by carrying out enzymatic hydrolysis by steps to obtain hydrolyzed whole egg liquid; step 3 in which the hydrolyzed whole egg liquid is mixed with prebiotics, then homogenized, sterilized, cooled, and inoculated with yeast and lactic acid bacteria to obtain whole egg fermentation liquid; and step 4 in which the whole egg fermentation liquid is mixed with cyclodextrin, then homogenized, entrapped, cooled at a low temperature environment and stood, and then centrifuged to remove cholesterol insoluble substances to obtain separation liquid which is then freeze-dried to finally obtain polypeptide-rich, high-calcium and low-cholesterol whole egg fermentation. The whole egg fermentation of the invention has stronger bioactivity, can produce positive influence on basic metabolism and immune system of human body, and can be widely applied to non-medical uses in the fields of nutritional functional foods, special foods and special medical foods, health foods and common foods. (Fig. 6) Fajiaocudanfen 1126 1255 RT18.31-40.00 AVA NL:8.50E5 T:FTMS+c Ed Full ms[205.00-15000] 649.29 to as 338.30 0 274.25 697.02 493.17 779.58 4s 0 is 360.37 so 318.3 20 I 45 . 2 5 786-17 ji 28.47 ~ ~28 s ,200 190 180 170 S160 150 140 *.130 ~120 0110 100 0 1 2 3 4 5 Week - Blank group -- fed with whole egg powder - fed with vinegar egg powder -- fed with whole egg fermentation FIG. 6 S3 AA

Description

Fajiaocudanfen 1126 1255 RT18.31-40.00 AVA NL:8.50E5 T:FTMS+c Ed Full ms[205.00-15000]

649.29 to

as

338.30 0

274.25 697.02 493.17 779.58 4s

is 360.37 so 318.3

20 I 45 . 2 5 786-17

ji 28.47 ~ ~28 s

,200 190 180 170 S160 150 140 *.130 ~120 0110 100 0 1 2 3 4 5 Week - Blank group -- fed with whole egg powder

- fed with vinegar egg powder -- fed with whole egg fermentation

FIG. 6

S3 AA

PRODUCTION METHOD OF HIGH BIOACTIVITY WHOLE EGG FERMENTATION, AND PRODUCTS AND APPLICATIONS THEREOF

FIELD The invention relates to the field of food and biological fermentation, and more particularly relates to a production method of high bioactivity whole egg fermentation, and products and applications thereof.

BACKGROUND Egg production in China ranks first in the world, while the egg processing industry lags behind. The factory is small in scale and quantity. The consumption of poultry eggs is mainly fresh eggs. However, fresh eggs have short shelf life and are easy to deteriorate, and eggshells are easy to be broken and inconvenient to transport, thus being inconvenient to directly apply in the food industry. Whole egg powder is an ideal substitute to fresh eggs. After fresh eggs are processed into egg powder, the egg powder has advantages of convenient use, long shelf life, no microbial pollution etc. and gets popular in the food processing industry. In the prior art, the process for preparing fresh eggs into whole egg powder mainly comprises two methods. One is a traditional method of soaking eggs with vinegar, and the products prepared thereby have the disadvantages such as great stimulation to intestines and stomach, inconducive to absorb biological macromolecular nutrients by human bodies, and of poor edibility. The other methods is to treat egg yolk and egg white merely, and the products prepared thereby have disadvantages of discarding eggshells which have high nutritive value, low utilization rate of nutritive substances, and low nutritive value.

SUMMARY An object of the present invention is to solve at least the above-mentioned problems and to provide at least advantages to be described later. Another object of the present invention is to provide a method for producing high bioactivity whole egg fermentation, and products and applications thereof. The fermentation products have stronger bioactivity and have a positive effect on human basic metabolism and immune system. In order to achieve these objects and other advantages according to the present invention, there is provided a method for producing high bioactivity whole egg fermentation, comprising: Step 1: poultry eggs or chicken embryo eggs are cleaned, sterilized and aired, and then submerged and soaked into edible organic acid of 1-3 times the weight of the poultry eggs for 1-6 days until eggshells are completely dissolved. The acid solution is filtered and collected. The egg membranes are rinsed with water to remove insoluble substances and pigment from the eggshells, then submerged again and soaked in the collected acid solution; Step 2: the egg is stirred to release the egg membrane and the egg liquid which are then subjected to continuous hydrolysis for 1-24 months under ultrasonic waves until the whole egg liquid is homogenized and free of lumps, and then were added therein lipase or phospholipase with the mass of 1/1000-1/100 times the mass of the egg and protease with the mass of 1/500-1/50 times the mass of the egg to continue enzymatic hydrolysis for 1-5 hours to obtain hydrolyzed whole egg liquid; Step 3: the hydrolyzed whole egg liquid is mixed with brown sugar and inulin prebiotics, then homogenized, sterilized, cooled, inoculated yeast and lactic acid bacteria, wherein the inoculation amount of the bacterial liquid is 1-5% of the volume of the hydrolyzed whole egg liquid, and finally fermented at 25-45°C for 1-15 days to obtain whole egg fermentation liquid; and Step 4: the whole egg fermentation liquid is filtered and added therein with cyclodextrin under ultrasonic waves, and then is dispersed at high speed, mixed for homogenization, embedded, cooled at a low temperature environment of 0-5°C and stood, and finally centrifuged to remove cholesterol insoluble substances. The separated liquid is freeze-dried to obtain whole egg fermentation. In the above method, the edible organic acid is selected from the group consisting of lactic acid, malic acid, butyric acid, acetic acid, edible rice vinegar, fruit vinegar and/or citric acid, and the concentration of the edible organic acid is 5-50wt%. In the above method, the lipase is derived from Aspergillus oryzae or Rhizopus oryzae, the phospholipase is selected from phospholipase Al, A2, C and/or D, and the protease is selected from bromelain, papain, ficin, pepsin, trypsin and/or acidic protease or neutral protease or alkaline protease derived from Aspergillus niger and Bacillus. In the above method, the yeast is Saccharomyces cerevisiae, and the lactobacillus is Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus reuteri,

Lactobacillus acidophilus and/or Propionibacterium fischeri. In the above method, the edible organic acid in step 1 may be subjected to a freezing-thawing pretreatment which comprises: the edible organic acid is placed in -20°C environment, into which is added an ice water mixture at 0°C with 0.5 times the weight of the edible organic acid for dissolution and is further stirred to form ice crystals, then therein another ice water mixture at 0°C with 0.5 times the weight of the edible organic acid is added for dissolution and further stirring to form ice crystals which is then placed in -20°C environment to freeze for 1 h and then mashed to fragments; the fragments are heated to 0°C at a rate of1°C/min and added with a further ice water mixture at 0°C to make the concentration of the edible organic acid reach 20wt%, and then is refrigerated for 1 h. The present invention further provides whole egg fermentation prepared by the above production method, of which the content of organic calcium is not less than 2wt%, the content of polypeptide protein is not less than 20wt%, the molecular weight distribution of polypeptide protein is concentrated between 0.5 and 3k, and the content ratio of small molecule polypeptide with molecular weight distribution concentrated below 1k is not less than 60wt%. The present invention also provides foods containing whole egg fermentation

, which comprises the whole egg fermentations and food ingredients or additives, which can be used as nutritional functional foods, special foods and special medical foods, health foods and non-medical uses in the food field. The application of the whole egg fermentation can be applied in preparing products for preventing hypertension and hyperlipidemia. The application of the whole egg fermentation can be applied in preparing products for increasing bone density, improving osteoporosis, enhancing physical strength, enhancing immunity, prolonging life and resisting aging. The application of the whole egg fermentation can be applied in preparing products for regulating menstruation, caring skin, moistening intestine, regulating constipation, improving intelligence and improving sleep. The invention at least comprises the following beneficial effects: According to the invention, acid hydrolysis, double enzyme hydrolysis, fermentation, embedding and other methods are adopted to not only improve the release of nutrients of whole eggs (eggshell, shell membrane, egg white and yolk liquid), but also improve the absorption utilization rate after biodegradation and conversion, and remove the eggy smell and excessive cholesterol of the original egg liquid and egg powder. The products are rich in a variety of active substances such as organic calcium, phospholipid type polypeptide oligopeptide, endogenous active enzymes and the like, and has higher antioxidant, blood pressure and lipid regulation, arteriosclerosis improvement, bone mineral density improvement, intelligence development and tranquilization, fatigue resistance, immunity improvement, menstruation regulation and beauty care and other biological activities compared with the original egg powder and vinegar egg powder and the like. The products can be widely applied to nutritional functional food, special food, special medical food, health food, etc. It can also further separate and purify related nutritional functional substances and be applied to non-medical applications in the fields of biomedicine and food. Other advantages, objects and features of the present invention will be reflected by the following description partly, and will be understood by those skilled in the art through research and practice of the present invention partly.

BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a diagram showing a result of ACE inhibition rate test; Fig. 2 is a diagram showing a result of ACE inhibition rate test; Fig. 3 shows a molecular weight distribution diagram of polypeptide; Fig. 4 shows a liquid chromatogram of polypeptide; Fig. 5 shows a liquid mass spectrum of polypeptide < 1kDa; Fig. 6 is a diagram showing a result of blood pressure test; Fig. 7 is a diagram showing a result of blood pressure test; and Fig. 8 is a diagram showing result of life extension test.

DETAILED DESCRIPTION The present invention will be described in further detail below with reference to examples to enable those skilled in the art to implement it with reference to the description. It should be understood that terms such as "comprise", "comprises" and "comprising" as used herein, do not exclude the presence or addition of one or more other elements or combinations thereof. It should be noted that the test methods in the following embodiments are conventional methods unless otherwise specified, and the reagents and materials are commercially available unless otherwise specified.

Example 1 A production method of high bioactivity whole egg fermentation comprises the following steps. Step 1: the raw material fresh poultry eggs were cleaned by tap water, and then cleaned by high-concentration electrolytic ozone water. The concentration of ozone in the cleaned water was 8 mg/l, and the water flow rate was 1201/h. The eggshell surface was sterilized, disinfected with 60% edible alcohol and aired. Then lactic acid having a weight twice of the poultry eggs and the concentration of 5 wt% is added so that the poultry eggs were submerged and soaked therein for 1 day until the eggshell was completely dissolved. The acid solution was filtered and collected. The egg membrane was rinsed by deionized water to remove insoluble substances and pigment of the eggshell, and then submerged and soaked in the collected acid solution again. Step 2: stirring the egg to release egg membrane and egg liquid, and the egg membrane and egg liquid are subjected to a continuous hydrolysis for 1 month under ultrasonic wave at a frequency of 100 Hz and lasting for 5 min until the whole egg liquid was homogenized and free of lumps, in which lipase with a mass of 1/1000 times of the egg and protease with a mass of 1/500 times of the egg were added to continue enzymatic hydrolysis for 1 hour to obtain hydrolyzed whole egg liquid, wherein the lipase was derived from Aspergillus oryzae, the phospholipase was selected from phospholipase Al and D, and the protease was selected from bromelain and papain. Step 3: the hydrolyzed whole egg liquid was mixed with 2wt% brown sugar and 5wt% inulin prebiotics, then homogenized, sterilized, cooled, inoculated with yeast and lactobacillus, wherein the inoculation amount of the bacterial liquid was 1% of the volume of the hydrolyzed whole egg liquid respectively, and finally fermented at 25°C environment for 1 day to obtain whole egg fermentation liquid, wherein the yeast was Saccharomyces cerevisiae, and the lactobacillus was Lactobacillus delbrueckii subsp. bulgaricus and Lactobacillus reuteri. Step 4: the whole egg fermentation liquid was filtered, added therein cyclodextrin at a volume mass ratio of 0.5% under ultrasonic waves at a frequency of 100 Hz and lasting for 5 min. The whole egg fermentation liquid was dispersed at high speed, mixed for homogenization, embedded, cooled at a low temperature environment of 0-5°C and stood, and finally centrifuged to remove cholesterol insoluble substances to obtain a separated liquid. The separated liquid was then freeze-dried to obtain whole egg fermentation.

Example 2 A production method of high bioactivity whole egg fermentation comprises the following steps. Step 1: the raw material chicken embryo eggs were cleaned by tap water, and then cleaned by high-concentration electrolytic ozone water, wherein the concentration of ozone in the cleaned water was 30 mg/l, and the water flow rate was 500 1/h. The eggshell surface was sterilized, disinfected with 80% edible alcohol and aired. Then malic acid and butyric acid having a weight three times of the chicken embryo eggs and a concentration of 50 wt% is added so that the poultry eggs were submerged and soaked therein for 6 days until the eggshell was completely dissolved. The acid solution was filtered and collected, whilst the egg membrane was rinsed by deionized water to remove insoluble substances and pigment of the eggshell, and then was once again submerged and soaked in the collected acid solution. Step 2: stirring the egg to release egg membrane and egg liquid, and the egg membrane and egg liquid are subjected to a continuous hydrolysis for 24 months under ultrasonic wave at a frequency of 500 Hz and lasting for 30 min until the whole egg liquid was homogenized and free of lumps, in which phospholipase having a mass of 1/100 times of the egg and protease having a mass of 1/50 times of the egg were added to continue enzymatic hydrolysis for 5 hours to obtain hydrolyzed whole egg liquid, wherein the lipase was derived from aspergillus oryzae, the phospholipase was phospholipase A2, and the protease was selected from ficin and pepsin. Step 3: the hydrolyzed whole egg liquid was mixed with 5 wt% brown sugar and 15 wt% inulin prebiotics, then homogenized, sterilized, cooled, inoculated with yeast and lactobacillus, wherein the inoculation amount of the bacterial liquid was 5% of the volume of the hydrolyzed whole egg liquid respectively, and is finally fermented at 45°C environment for 15 days to obtain whole egg fermentation liquid, wherein the yeast was Saccharomyces cerevisiae, and the lactobacillus was selected from Lactobacillus acidophilus and Propionibacterium fischeri. Step 4: the whole egg fermentation liquid was filtered, added cyclodextrin with a volume mass ratio of 15% under ultrasonic waves at a frequency of 500 Hz and lasting for 30 min. The whole egg fermentation liquid was dispersed at high speed, mixed for homogenization, embedded, cooled at a low temperature environment of 0-5°C and stood, and finally centrifuged to remove cholesterol insoluble substances to obtain a separated liquid. The separated liquid was freeze-dried to obtain a whole egg fermentation.

Example 3 A production method of high bioactivity whole egg fermentation comprises the following steps. Step 1: the raw material poultry eggs were cleaned by tap water, and then cleaned by high-concentration electrolytic ozone water, wherein the concentration of ozone in the cleaned water was 20 mg/l, the water flow rate was 300 1/h. The eggshell surface was sterilized, disinfected with 70% edible alcohol and aired, then added therein edible rice vinegar having twice the weight of the poultry eggs and a concentration of 20wt%. The poultry eggs were submerged and soaked for 3 days until the eggshell was completely dissolved. The acid solution was filtered and collected and the egg membrane was rinsed by deionized water to remove insoluble substances and pigment of the eggshell, and then was submerged and soaked in the collected acid solution again. Step 2: stirring the egg to release egg membrane and egg liquid, which are subjected to continuous hydrolysis for 12 months under ultrasonic wave at a frequency of 300 Hz and lasting for 15 min until the whole egg liquid was homogenized and free of lumps, in which lipase with a mass of 1/500 times of the egg and protease with a mass of 1/300 times of the egg were added to continue enzymatic hydrolysis for 3 hours to obtain hydrolyzed whole egg liquid, wherein the lipase was derived from Rhizopus oryzae, the phospholipase was phospholipase C, and the proteases were acidic proteases derived from Aspergillus niger and Bacillus. Step 3: the hydrolyzed whole egg liquid was mixed with 4wt% brown sugar and l0wt% inulin prebiotics, then homogenized, sterilized, cooled, inoculated with yeast and lactobacillus, wherein the inoculation amount of the bacterial liquid was 3% of the volume of the hydrolyzed whole egg liquid respectively, and finally fermented at 35°C environment for 8 days to obtain whole egg fermentation liquid, wherein the yeast was Saccharomyces cerevisiae, and the lactobacillus was selected from Lactobacillus acidophilus and Propionibacterium fischeri. Step 4: the whole egg fermentation liquid was filtered, added therein cyclodextrin having a volume mass ratio of 10% under ultrasonic waves at a frequency of 300 Hz and lasting for 15 min. The whole egg fermentation liquid was dispersed at high speed, mixed for homogenization, embedded, cooled at a low temperature environment of

0-5°C and stood, and finally centrifuged to remove cholesterol insoluble substances to obtain a separated liquid. The separated liquid was then freeze-dried to obtain whole egg fermentation.

Example 4 A production method of high bioactivity whole egg fermentation comprises the following steps. Step 1: the raw material chicken embryo eggs were cleaned by tap water, and then cleaned by high-concentration electrolytic ozone water, wherein the concentration of ozone in the cleaned water was 20 mg/l, the water flow rate was 300 1/h. The eggshell surface was sterilized, disinfected with 60-80% edible alcohol and aired, then added therein edible rice vinegar having a weight twice of the chicken embryo eggs and a concentration of 20wt%, in which the poultry eggs were submerged and soaked for 3 days until the eggshell was completely dissolved. The acid solution was filtered and collected and the egg membrane was rinsed by deionized water to remove insoluble substances and pigment from the eggshell, and then was submerged and soaked in the collected acid solution again. Additionally, the edible organic acid in step 1 was subjected to a freezing-thawing pretreatment, which comprises that the edible rice vinegar was placed in -20°C environment, into which was added an ice water mixture at 0°C with 0.5 times the weight of the edible rice vinegar to form ice crystals by stirring, then was added therein another ice water mixture at0°C with 0.5 times the weight of the edible organic acid to form ice crystals by stirring and was placed in -20 0C environment to freeze for 1 h and then mashed to fragments. The fragments were heated to 0°C at a rate of1°C/min and added therein a further ice water mixture at 0°C,

and then refrigerated for 1 h to make the concentration of the edible organic acid reach 20wt%. Step 2: stirring the eggs to release egg membrane and egg liquid, and making the egg membrane and egg liquid continue to hydrolysis for 12 months under ultrasonic wave at a frequency 300 Hz and lasting for 15 min until the whole egg liquid was homogenized and free of lumps, in which lipase with a mass of 1/500 times of the egg and protease with a mass of 1/300 times of the egg were further added to continue enzymatic hydrolysis for 3 hours to obtain hydrolyzed whole egg liquid, wherein the lipase was derived from Rhizopus oryzae, the phospholipase was phospholipase C, and the proteases were acidic proteases derived from Aspergillus niger and Bacillus. Step 3: the hydrolyzed whole egg liquid was mixed with 4wt% brown sugar and l0wt% inulin prebiotics, then homogenized, sterilized, cooled, inoculated with yeast and lactobacillus, wherein the inoculation amount of the bacterial liquid was 3% of the volume of the hydrolyzed whole egg liquid respectively, and finally fermented at 35°C environment for 8 days to obtain whole egg fermentation liquid, wherein the yeast was Saccharomyces cerevisiae, and the lactobacillus was selected from Lactobacillus acidophilus and Propionibacterium fischeri. Step 4: the whole egg fermentation liquid was filtered, added therein cyclodextrin with a volume mass ratio of 10% under ultrasonic wavesat a frequency of 300 Hz and lasting for 15 min. The whole egg fermentation liquid was dispersed at high speed, mixed for homogenization, embedded, cooled at a low temperature environment of 0-5°C and stood, and finally centrifuged to remove cholesterol insoluble substances to obtain a separated liquid. The separated liquid was freeze-dried to obtain whole egg fermentation.

Example 5 The whole egg fermentation prepared in Example 3 was prepared to make cakes with egg liquid, granulated sugar, cake oil, salt, water, low gluten flour and bubble water.

Example 6 The whole egg fermentation prepared in Example 3 was prepared to make meal replacement powder with filler, adsorbent, binder, lubricant, dispersant, wetting agent, disintegrant, perfume and pigment.

Example 7 The whole egg fermentation prepared in Example 3 was prepared to make effervescent tablets with diluent, binder, disintegrant and lubricant.

Example 8 The whole egg fermentation prepared in Example 4 was filled into soft capsule body with dispersant, plasticizer, disintegrant, opacifier and lubricant, and soft capsules can be prepared by pelleting, shaping and drying, pill washing, pill airing and pill picking.

Evaluation Experiments on Efficacy of Pretreatment using Edible Organic Acid 1. Effect of Pretreatment using Edible Organic Acid on Extraction of Egg Keratin

The poultry eggs were aired, added edible rice vinegar and soaked for 3 days until the egg shells were completely dissolved, wherein the weight of the edible rice vinegar was twice the weight of the poultry eggs and had a concentration of 20wt%, submerging and soaking the egg for 3 days until the egg shells were completely dissolved. The acid solution was filtered and collected, and the egg membranes were rinsed with water. Insoluble substances and pigment from the eggshells were removed. The egg membrane was left behind, washed and weighed to get an initial mass, and finally submerged again and soaked in the collected acid solution. The dissolution time of the egg membrane was calculated with a time at which the egg membrane was just completely dissolved and no visible fragments exist being an end point. The solution was concentrated to 100 ml at room temperature. The pH was adjusted to isoelectric point to precipitate and separate keratin. The keratin was allowed to stand for 30 min. The insoluble substance was extracted again by centrifugation. The lower layer keratin was transferred to a dialysis bag for dialysis for 3 days and water was changed once every 4 h, wherein the dialysis bag had a diameter of 25 mm, a molecular weight cut-off of 8000-14000 D and was made of regenerated cellulose material. After dialysis was completed, the purified keratin was vacuum freeze-dried at a vacuum pressure of 15 Pa and a temperature of -45°C to obtain keratin. The keratin was weighed to obtain keratin mass, and the keratin yield was calculated as: keratin yield = keratin mass/egg membrane initial mass x 100%.

Comparative Example 1 The edible rice vinegar was edible rice vinegar at room temperature.

Comparative Example 2 The edible rice vinegar was edible rice vinegar undergone a freezing-thawing pretreatment, and specifically comprises the following steps. The edible organic acid was placed in -20°C environment, into which was added an ice water mixture at 0°C with 0.5 times the weight of the edible organic acid to dissolve and obtain a mixture A; the mixture A was stirred to form ice crystals and was added therein another ice water mixture at 0°C with 0.5 times the weight of the edible organic acid to dissolve and obtain a mixture B; the mixture B was stirred to form ice crystals and was placed in -20°C environment to freeze for 1 h and then mashed to fragments; the fragments were heated to 0C at a rate of

1°C/min and added a further ice water mixture at 0°C, and then refrigerated for 1 h to make the concentration of the edible organic acid reach 20wt%. Table 1 Dissolution time of egg Keratinyield membrane (min)

Comparative example 1 88 11.34

Comparative example 2 45 27.82

The results shows that the dissolution time of egg membrane in comparative example 2 was significantly shorter than that in comparative example 1, while the keratin yield was significantly higher than that in comparative example 1. This is because edible rice vinegar undergoes a freezing-thawing pretreatment, which could swell egg membrane at a faster speed, break hydrogen bonds and salt bonds on the surface of egg membrane, and release keratin embedded in egg membrane.

2. Effect of Pretreatment using Edible Organic Acid on Amino Acid Extraction of Protein The poultry eggs were aired, added edible rice vinegar and soaked for 3 days until the egg shells were completely dissolved, wherein the weight of the edible rice vinegar was twice the weight of the poultry eggs and the concentration was 20wt%, submerging and soaking the egg for 3 days until the egg shells were completely dissolved. The acid solution was filtered and collected, and the egg membranes were rinsed with water. Insoluble substances and pigment from the eggshells were removed. The egg membrane was submerged again and soaked in the collected acid solution. Stirring the egg to release egg membrane and egg liquid, which are subjected to continuous hydrolysis for 12 months under ultrasonic wave at a frequency of 300 Hz and lasting for 15 min until the whole egg liquid was homogenized and free of lumps, into which Protease having a mass of 1/300 times of the egg was added to continue enzymatic hydrolysis for 3 hours to obtain hydrolyzed whole egg liquid, wherein the lipase was derived from Rhizopus oryzae, the phospholipase was phospholipase C, and the proteases were acidic proteases derived from Aspergillus niger and Bacillus; The hydrolyzed whole egg liquid was mixed with 4wt% brown sugar and 10wt% inulin prebiotics, then was homogenized, sterilized, cooled, inoculated yeast and lactobacillus, wherein the inoculation amount of the bacterial liquid was 3% of the volume of the hydrolyzed whole egg liquid respectively, and finally fermented at 35°C environment for 8 days to obtain whole egg fermentation liquid, wherein the yeast was Saccharomyces cerevisiae, and the lactobacillus was selected from Lactobacillus acidophilus and Propionibacterium fischeri. The whole egg fermentation liquid was filtered and homogenized. The amino acid content (mg/100 ml) was determined by GB/T 5009.124-2003.

Comparative Example 3 The edible rice vinegar was edible rice vinegar at room temperature.

Comparative Example 4 The edible rice vinegar was edible rice vinegar already subjected to a freezing-thawing pretreatment, and specifically comprises the following steps. The edible organic acid was placed in -20°C environment, into which was added an ice water mixture at 0°C and having 0.5 times the weight of the edible organic acid to dissolve and obtain a mixture A; the mixture A was stirred to form ice crystals and was added another ice water mixture at 0°C having 0.5 times the weight of the edible organic acid to dissolve and obtain a mixture B; the mixture B was stirred to form ice crystals and was placed in -20°C environment to freeze for 1 h and then mashed to fragments; the fragments were heated to 0°C at a rate of1C/min and added a further ice water mixture at0°C, and then refrigerated for 1 h to make the concentration of the edible organic acid reach 20wt%. Table 2

Cmp. Cmp. Cmp. Cmp. example 3 example 4 example 3 example 4

Detected, Detected, Detected, Detected, Aspartate Methionine* 16.77 17.11 6.88 7.24

Detected, Detected, Detected, Detected, Tyr Isoleucine* 6.89 8.32 11.16 12.21

Detected, Detected, Detected, Detected, Serine Leucine* 16.26 18.91 16.68 21.58

Detected, Detected, Detected, Detected, Glutamate Threonine* 27.92 36.67 13.45 14.52

Proline Not detected Detected, Phenylalanine* Detected, Detected,

21.55 6.99 8.27

Detected, Detected, Detected, Detected, Glycine 12.68 14.25 Lysine* 26.78 27.15

Detected, Detected, Not Detected, Alanine Valine* 22.52 24.11 detected 14.26

Detected, Detected, Detected, Cystine Not detected 2.67 Tryptophan* 4.76 5.33 2.67 4.76 5.33

Detected, Detected, Not Detected, Arginine Histidine* 8.59 9.22 detected 9.23

The results showed that 18 kinds of amino acids were detected in comparative example 4 and 14 kinds of amino acids were detected in comparative example 3. This is because edible rice vinegar can release keratin in egg membrane after freezing-thawing pretreatment, which enhances bioactivity and bioavailability.

Efficacy Evaluation Experiments of Whole Egg Fermentation Comparative Example 5 A production method of whole egg powder which is the same as in the prior art and a reference can be made to Study on New Technology of Industrialized Production of

Special Chicken Whole Egg Powder by Lou Yuangong, and hereby was briefly described as follows: raw material fresh eggs -- sensory examination -- cleaning -- disinfection with shell -- air the eggs -- semi-automatic egg stirring (shell discarding) -- filtration (residue discarding) -- preparing -- sterilization -- spray drying -- sieving -- inspection

and packaging -- finished product.

Comparative Example 6 A production method of vinegar egg powder which is the same as in the prior art and a reference can be made to Study on the Change of Protein in Vinegar Egg Soaking

and the Preparation of Vinegar Egg Powder by Wang Danbo, and hereby was briefly described as follows: eggs -- washing, disinfection (pretreatment) -- vinegar soaking - stirring evenly -- pressure filtration -- vinegar egg liquid -- disinfection -- spray drying -- vinegar egg powder -- packaging.

1 Anti-aging Experiments 1.1 Animal Experiments Seventy male Kunming mice of 18-22 g were selected and fed for one week. They were randomly divided into blank control group, model group, whole egg powder group, vinegar egg powder group, low-dose group, medium-dose group and high-dose group with 10 mice in each group. The blank control group received subcutaneous injection of 300 mg/(kg-d) normal saline every day. The model group, whole egg powder group, vinegar egg powder group, low-dose group, medium-dose group and high-dose group were injected with D-galactose 300 mg/(kg-d) subcutaneously every day. The blank control group and the model group was fed normal saline by gavage at a dose of 150 mg/(kg-d), the whole egg powder group was fed whole egg powder prepared in comparative example 5 by gavage at a dose of 150 mg/(kg-d), the vinegar egg powder group was fed vinegar egg powder prepared in comparative example 6 by gavage at a dose of 150 mg/(kg-d), and the high-dose group, the medium-dose group and the low-dose group was fed whole egg fermentation prepared in example 3 by gavage at a dose of 300 mg/(kg-d),150 mg/(kg-d) and 20 mg/(kg-d) respectively once a day for 4 weeks. Blood was taken by eyeball enucleation and liver was taken after perfusing with normal saline. The activity of total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-Px) and the content of malondialdehyde (MDA) in serum and liver tissues of mice were detected according to the use method of the kit, and the results are shown in Table 3. Table 3

T-SOD activity GSH-Px MDA content (U/i) activity (U/) (nmol/ml)

Blank control group 98.14+4.66 338.49+29.19 2.31+0.36

Model group 45.49+13.94 188.21+22.83 4.29+0.27

Whole egg powder group 48.96+10.47 194.28+19.29 4.18+0.34

Vinegar egg powder group 54.47+8.24 227.04+9.12 3.63+0.52

Low-dose group of whole egg fermentation 47.87+11.51 195.85+21.51 4.19+0.72

Medium-dose group of whole egg fermentation 77.39+15.22 211.60+24.39 3.27+0.48

High-dose group of whole egg fermentation 92.03+12.36 279.68+32.30 2.78+0.31

Analysis of test results: Compared with the blank control group, the activities of T-SOD and GSH-Px in the serum and liver of the model group decreased significantly, and the MDA content increased significantly. The difference was very significant, indicating that the aging mouse model was successfully established. It can be seen from Table 3 that compared with the model group, the high-dose group and the medium-dose group can significantly increase the activity of T-SOD in serum and significantly reduce the content of MDA. The GSH-Px activity in serum was significantly increased in high-dose group, medium-dose group and low-dose group. The high-dose group was significantly higher than the low-dose group in raising T-SOD in mice serum, and the GSH-Px enzyme activity was significantly higher than that of the medium-dose group and the low-dose group. For reducing MDA level, there was no significant difference among high-dose group, medium-dose group and low-dose group. Compared with the whole egg powder group and vinegar egg powder group, the high-dose group was significantly higher than the whole egg powder group and vinegar egg powder group in raising T-SOD in mice serum, and the GSH-Px enzyme activity was significantly higher than the whole egg powder group and vinegar egg powder group. As can be seen from Table 3, compared with the model group, the high-dose group, the medium-dose group and the low-dose group can significantly enhance the enzyme activities of T-SOD and GSH-Px in the liver and significantly reduce the MDA content. The activities of T-SOD and GSH-Px in liver of high-dose mice were significantly higher than those of low-dose mice. MDA content in high-dose group was significantly lower than that in low-dose group. Compared with the whole egg powder group and the vinegar egg powder group, the activity levels of T-SOD and GSH-Px in the liver of the mice in the high-dose group were higher than those in the whole egg powder group and the vinegar egg powder group, and the MDA content in the liver was lower than those in the whole egg powder group and the vinegar egg powder group, which indicated that the high-dose group of the whole egg fermentation was superior to the whole egg powder group and the vinegar egg powder group in improving the antioxidant activity in vivo.

1.2 Human Body Experiments

100 people were selected and randomly divided into a drug administration group and a blank control group, with 50 people in each group, 20 men and 30 women in the drug administration group, with the minimum age of 46 years, the maximum age of 64 years and an average age of 54.32 years. The blank control group had 15 men and 35 women, with the minimum age of 45 years and the maximum age of 62 years, with an average age of 53.14 years. The drug administration group took 7.5 g of the whole egg fermentation prepared in Example 3 orally every day for 4 consecutive weeks. The blank control group did not take any drugs, and the indexes of superoxide dismutase (SOD) and malondialdehyde (MDA) were measured respectively. The results are shown in Table 4. Table 4 Time (weeks) Indicators Experiment group TO TI T2 T3 T4

SOD Blank control group 98.24+10.32 96.72+11.31 97.23+15.46 96.63+17.21 96.98+12.74 (U/ml) Administration group 102.31+12.57 105.2810.63 106.18+9.83 110.24+13.22 112.23+9.46

MDA Blank control group 7.59+0.94 7.26+1.13 7.41+0.83 7.63+1.02 7.55+.0.92 nmol-ml Administration group 7.32+0.78 6.92+0.82 6.76+0.65 6.28+0.47 5.85+0.83

Analysis of test results: There was no significant difference in superoxide dismutase (SOD) and malondialdehyde (MDA) between the administration group and the blank control group at week 0. At the 4th week in the administration group, the serum SOD level increased significantly from 102.3112.57 U/ml to 112.239.46 U/ml. The level of serum MDA decreased significantly from 7.32 0.78 nmol/ml to 5.85 0.83 nmol/ml. The results of routine blood tests, stool and urine tests and blood biochemical tests in the test group showed no obvious abnormalities and no allergic reactions.

2 Enhance Physical Strength and Immunity Experiments 2.1 Physical Strength Enhancement Experiments Seventy male Kunming mice of 18-22 g were selected and fed for one week. They were randomly divided into blank control group, whole egg powder group, vinegar egg powder group, low-dose group, medium-dose group, high-dose group and preferred medium-dose group, with 10 mice in each group.

The blank control group was fed distilled water by gavage at a dose of 150 ml/(kg-d) daily, the whole egg powder group was fed with the whole egg powder prepared in comparative example 5 at a dose of 150 mg/(kg-d), the vinegar egg powder group was fed with the vinegar egg powder prepared in comparative example 6 at a dose of 150 mg/(kg-d), and the high-dose group, the medium-dose group and the low-dose group were fed the whole egg fermentation prepared in example 3 at a dose of 300 mg/(kg-d), 150 mg/(kg-d) and 20 mg/(kg-d) respectively. The preferred medium-dose group was fed with the whole egg fermentation prepared in Example 4 at a dose of 150 mg/(kg-d), once a day for 30 consecutive days. The above groups of mice underwent a weight-bearing swimming Experiments to detect swimming time, liver glycogen, lymphocyte A value, serum hemolysin antibody mean value and red blood cell immune function, and the test results were shown in Table 6; Spleen lymphocyte transformation Experiments induced by concanavalin A (ConA), namely MTT method, is adopted as method for detecting lymphocyte A value. Spleen was taken under aseptic conditions and spleen cell suspension was prepared with complete culture medium. 2 ml of spleen cell suspension was inoculated on cell culture plates respectively. 50 pL of ConA solution with a concentration of 100 mg/l was added, another was not added with ConA as a control. The spleen cell suspension was cultured in a C02 incubator at 37°C until 68 h later was added 100 [LMTT solution (50 mg/l). After continuous culture for 4 h, acidic isopropanol was added to terminate the reaction, and A value was measured with E960 microplate reader. Methods for detecting the mean value of serum hemolysin antibody: Blood was taken by eyeballs enucleation. Serum was centrifuged, diluted into double volume with normal saline, placed in a 90-hole U-shaped hemagglutination plate for incubation respectively. The hemagglutination reaction results were observed. Methods of erythrocyte immune function: Blood was collected from the fundus venous plexus of mice, and the white blood cell (WBC) content was measured by Japanese CC-180 automatic blood cell counter. One drop of mouse fundus venous plexus blood was taken from 2 ml of normal saline, washed and centrifuged for 3 times at a rotation speed of 1000 rpm/5 min. Red blood cell suspension of 1.25x10 / was prepared with normal saline, and 5 drops of red blood cell suspension and C 3 b sensitized yeast suspension (1x10 1 1 /1) or non-sensitized yeast suspension (1x10" /1) were taken from a syringe respectively, and fully mixed in a test tube, incubated in a 37°C water bath for 30 min, added with 10 drops of normal saline, diluted and mixed evenly, and then fixation was done with 2 drops of 0.25% defensineal. After mixing evenly, the smears were stained with Wright's stain and observed with wet smears. 200 red blood cells were counted at high magnification. Combined with the number of rosette cells of more than 2 yeasts, RBC-C3b receptor rosette rate, i. e., sensitized yeast suspension, and RBC-IC rosette rate, i. e., non-sensitized yeast suspension were calculated,. Table 5

Liver Lymphocyte A RBC adhesion glycogen value Serum function Experiment Swimming content hemolysin RBC-C3b WBC RCCbRBC-ICG group time (min) (mg/loo With Without (antibody Receptor ConA ConA mean) rosette rosette mg liver tissue) rate rate

Blank control 1545.63± 0.72± 45.23±8.32 0.64±0.25 0.89±0.50 10.7±2.53 4.32±2.01 8.20±1.01 group 273.45 0.04

Whole egg 1602.49± 0.68± 60.14±7.42 0.73±0.16 0.93±0.13 10.7±1.73 4.19±2.33 8.32±1.28 powder group 192.57 0.05

Vinegar egg 1734.82± 0.81± 69.35±11.70 0.86±0.23 1.02±0.45 11.5±2.35 4.35±1.97 7.46±1.33 powder group 225.61 0.10

Low-dose group of 1698.53± 0.92± 73.24±10.65 0.75±0.14 1.00±0.36 10.8±1.83 4.21±2.22 7.73±0.95 whole egg 198.28 0.13 fermentation

Medium-dose group of 1858.72± 1.18± 95.61±9.73 0.93±0.63 1.49±0.23 13.5±2.94 5.94±1.92 5.58±2.03 whole egg 312.14 0.24 fermentation

High-dose group of 2314.67± 1.46± 115.58+10.83 1.09±0.37 1.68±1.39 14.7±1.32 6.82±2.36 4.52±1.36 whole egg 247.36 0.11 fermentation

Preferred 2169.23± 1.30± 0.98±0.31 1.52±0.39 14.1±2.02 6.21±2.77 5.11±1.76 medium-dose 102.81±6.73 231.43 0.19 group

Analysis of test results: Compared with the blank control group, the weight-bearing swimming time of the mice was obviously prolonged in the preferred medium-dose group, low-dose group, medium-dose group and high-dose group. The results of liver glycogen content Experiments in mice showed significant differences in the preferred medium-dose group, low-dose group, middle dose group and high-dose group compared with blank control group. For proliferation reaction of mouse spleen lymphocytes, the results showed that the lymphocyte A values of the preferred medium-dose group, low-dose group, medium-dose group and high-dose group were significantly higher than that of the blank control group regardless of whether ConA was added or not, and the high-dose group was the most significant. The determination results of serum hemolysin content in mice showed that the antibody numbers in the preferred medium-dose group, low-dose group, medium-dose group and high-dose group were higher than those in the blank control group with significant difference. Glycogen is an important source of energy during exercise. Increasing the storage amount of glycogen is helpful to enhance endurance and exercise ability and to resist fatigue and improve the immune level of the body. Compared with the blank control group, the WBC and RBC-C 3b receptor rosette rates were significantly increased and the RBC-IC rosette rates were significantly decreased in the preferred medium-dose group, medium-dose group and high-dose group, which indicated that the preferred medium-dose group, medium-dose group and high-dose group could resist the generation of fatigue and improve the cellular immune andhumoral immune functions of mice.

2.2 Immunity Enhancement Experiments Eighty male Kunming mice of 18-22 g were selected and were randomly divided into blank control group, model group, whole egg powder group, vinegar egg powder group, low-dose group, medium-dose group, high-dose group and preferred medium-dose group after feeding for one week, with 10 mice in each group. The blank control group was fed with 150 mg/(kg-d) distilled water daily, and the model group, whole egg powder group, vinegar egg powder group, low-dose group, medium-dose group, high-dose group, and preferred medium-dose group were fed with 75 mg/(kg-d) cyclophosphamide (CY) normal saline subcutaneously daily. The whole egg powder group was fed whole egg powder prepared in comparative example 5 by gavage at a dose of 150 mg/(kg-d),the vinegar egg powder group was fed vinegar egg powder prepared in comparative example 6 by gavage at a dose of 150 mg/(kg-d), the high-dose group, the medium-dose group and the low-dose group was fed whole egg fermentation prepared in example 3 by gavage at a dose of 300 mg/(kg-d), 150 mg/(kg-d) and 20 mg/(kg-d) respectively, and the preferred medium-dose group was fed with the whole egg fermentation prepared in example 4 by gavage at a dose of 150 mg/(kg-d). Each group was fed by gavage once a day.

2.21 Effect on Phagocytosis of immunosuppressive mouse peritoneal macrophages Induced by Cyclophosphamide CY After continuous treatment for 7 days, on the 8th day, each group of mice were intraperitoneally injected with 5% chicken red blood cell normal saline suspension. After 6 h, the mice were killed, the abdomen was disinfected, 2.5ml of Hansen's solution was injected, the abdomen of the mice were gently rubbed, the abdominal cavity solution was placed on a slide, wet gauze was laid under the slide, and the abdominal cavity solution was incubated in an incubator at 37C for 30 min. The floating cells were washed away by normal saline, and Wright's dye solution was used for dyeing. Then the cells were washed, dried and observed under a microscope, and the phagocytic percentage and phagocytic index were calculated. The results are shown in Table 6.

2.22 Effect on Hemolysin Formation in Immunosuppressed Mice Induced by Cyclophosphamide CY On the first day of gavage, mice in each group were injected intraperitoneally with 0.2 ml of 5% chicken red blood cell normal saline suspension for immunization. On the 8th day, blood was taken from the orbit, serum was centrifuged, diluted with normal saline at a volume ratio of 1:100, 1 ml of which was taken, mixed with 0.5 ml of 5% chicken red blood cell suspension and 0.5 ml of 10% complement, and incubated at 37C for 30 min. Ice water was used to stop the reaction, supernatant was taken for color comparison at 540 nm with 72-1 spectrophotometer, and blank tubes without complement were used as controls. The results are shown in Table 6.

2.23 Effect on Hemolytic Plaque Formation in Immunosuppressed Mice Induced by Cyclophosphamide CY On the first day of gavage, each group of mice was injected intraperitoneally with 0.2 ml of 5% chicken red blood cell normal saline suspension for immunization. On the 8th day, after gavage the spleen was taken and the spleens of three mice were put together and homogenized with a homogenizer to adjust the concentration of spleen cells to 5x106 cells /ml. 0.5 ml of spleen cell suspension was added 0.2% chicken red blood cell suspension and 0.5 ml of 1:10 guinea pig serum respectively, mixed evenly, incubated at 37°C for 1 h, centrifuged, and the supernatant was taken for color comparison with 72-1 spectrophotometer , and zeroed with a blank tube without complement.

2.24 Effect on T Cell Percentage and Lymphocyte Transformation in Peripheral Blood of Immunosuppressed Mice Induced by Cyclophosphamide CY On the 1st to 3rd day of gavage administration, each group of mice were injected with 10 mg/kg PHA intramuscularly. After 2 h of gavage administration on the 8th day, the mice tails were cut off to take blood, blood sample was adopted, dried by natural wind and immersed in an incubation solution preheated for 30 min to incubate for 3 h. Sediment was removed with water, water droplets was absorbed by filter paper, the sample was dyed with 1% malachite green, then washed and dried, and the percentage of T lymphocytes was calculated with oil lens. Blood were taken from mice and smeared, Wright staining was performed, and lymphocyte transformation rate was calculated by oil lens. The results are shown in Table 6. Table 6 Inhibit the formation of Inhibition of T cell Inhibitionofmacrophage hemolysin and percentage and lymphocyte phagocytosis hemolytic plaque transformation rate

Experiment group Percentage Hemolytic Transformation Hemolysin Percentage of Phagocytosis plaque rate of formation plqe of peripheral phagocytosis index formation drenched cells (n=10)(OD) T cells(%o) (%) (n=3)(OD) (%)

Blank control group 34.34+7.02 0.74+0.06 0.14+0.09 0.52+0.03 51.37+3.14 43.09+3.82

Model group 22.67+6.17 0.34+0.10 0.03+0.02 0.34+0.01 27.42+5.03 21.37+1.56

Whole egg powder 36.46+8.35 0.38+0.12 0.03+0.03 0.33+0.02 29.65+4.33 24.52+2.47 group

Vinegar egg powder 42.36+5.46 0.63+0.08 0.04+0.01 0.42+0.03 37.56+6.67 35.47+2.61 group

Low-dose group of whole egg 32.34±5.67 0.48+0.11 0.03+0.03 0.37+0.01 28.83+3.17 23.85+3.27 fermentation

Medium-dose group of whole egg 45.36+6.42 0.66+0.09 0.04+0.02 0.44+0.03 47.25+6.42 40.34+2.33 fermentation

High-dose group of whole egg 63.67+4.93 0.82+0.15 0.07+0.03 0.52+0.02 62.14+4.25 49.62+3.45 fermentation

Preferred medium-dose group 50.56+2.34 0.71+0.12 0.05+0.01 0.50+0.02 50.23+5.89 42.77+3.08 of whole egg fermentation

Analysis of test results: Phagocytosis percentage and phagocytic index in model group were significantly reduced. Compared with model group, low-dose group, medium-dose group, high-dose group and preferred medium-dose group all significantly increased phagocytic percentage and phagocytic index of immunosuppressive mouse peritoneal macrophages Immunosuppressive mouse peritoneal macrophages. The hemolysin and OD values of hemolytic plaque formation in the model group were significantly lower than those in the blank control group. Compared with the model group, the high-dose group, preferred medium-dose group, can significantly promote the formation of hemolysin and hemolytic plaque. The percentage of peripheral T cells and the transformation rate of lymphocytes in the model group were significantly lower than those in the blank control group. Compared with the model group, the percentage of peripheral T cells and the transformation rate of lymphocytes in immunosuppressed mice were significantly increased in the medium-dose group, the high-dose group and the preferred medium-dose group. The above results show that the whole egg fermentation has a significant promotion effect on the multi-link immune function, can promote the formation of hemolysin and hemolytic plaque, promote the transformation of lymphocytes and increase the percentage of peripheral T cells, and significantly resist the immunologic hypofunction caused by cyclophosphamide.

3. Intelligence Experiments 3.1 Dark Avoidance Experiments Fifty male Kunming mice of 18-22 g were selected and fed for one week. They were randomly divided into blank control group, whole egg powder group, vinegar egg powder group, whole egg fermentation group and preferred medium-dose group with 10 mice in each group. The blank control group was fed with 150 mg/(kg-d) of distilled water by gavage, the whole egg powder group was fed with the whole egg powder prepared in comparative example 5 by gavage at a dose of 150 mg/(kg-d), the vinegar egg powder group was fed with the vinegar egg powder prepared in comparative example 6 at a dose of 150 mg/(kg-d), and the whole egg fermentation group was fed with whole egg fermentation prepared in example 3 by gavage at a dose of 150 mg/(kg-d), and the preferred medium-dose group was fed with the whole egg fermentation prepared in Example 4 by gavage at a dose of 150 mg/(kg-d). The mice were fed once a day for 30 days continuously, and the training was started at the 31st day. During the training, the same dose was continuously fed by gavage once a day. During the Experiments, the animal was placed in the bright room with its face facing backwards the hole, and the timer was started at the same time. If the animal passed through the hole and entered the dark room, it will be shocked, the timer will automatically stop, and the mouse will be taken out. The time required for each animal to enter the dark room from the bright room to avoid electric shock was recorded, i. e., the incubation period. The Experiments were repeated 24 or 48 hours later, and the number of times each animal entered the dark room in 10 min, i. e., the number of errors, was recorded. The results are shown in Table 7. Table 7

Experiment group Number of errors

Blank control group 6.8±2.2

Whole egg powder group 6.3+2.4

Vinegar egg powder group 3.8±1.5

Whole egg fermentation group 2.9+2.4

Preferred whole egg fermentation group 2.1+2.6

Analysis of test results: Compared with the blank control group, the whole egg fermentation group and the preferred medium-dose group has significantly reduced the number of times of entering the dark room, that is, the number of mouse errors; Compared with the whole egg powder group and vinegar egg powder group, the whole egg fermentation group and the preferred medium-dose group significantly reduces the number of mistakes in mice.

3.2 Water Maze Experiments Fifty male Kunming mice of 18-22 g were selected and fed for one week. They were randomly divided into blank control group, whole egg powder group, vinegar egg powder group, whole egg fermentation group, preferred medium-dose group, with 10 mice in each group. The blank control group was fed with 150 mg/(kg-d) of distilled water by gavage, the whole egg powder group was fed with 150 mg/(kg-d) of the whole egg powder prepared in comparative example 5 by gavage, the vinegar egg powder group was fed with 150 mg/(kg-d) of the vinegar egg powder prepared in comparative example 6 by gavage, and the whole egg fermentation group was fed with 150 mg/(kg-d) of the whole egg fermentation prepared in example 3 by gavage. The preferred medium-dose group was fed with the whole egg fermentation prepared in Example 4 at a dose of 150 mg/(kg-d) once a day for 30 days by gavage. The training starts at the 31st day. During the training, the same dose was continuously fed once a day by gavage. The water depth in the maze lane was 15 cm, the water temperature was 25°C, the time limit was 120s, all animals that do not reach the destination within 120s were recorded as 120s. During the Experiments, the animals were placed at the starting point of the lane, and the start button was pressed at the same time. The time when the animals reached the end point and the number of animals that reached the end point was recorded, and were shown in Table 8. Table 8

Experiment group Time required for maze (s)

Blank control group 93.6+14.2

Whole egg powder group 84.9+17.8

Vinegar egg powder group 78.6+11.2

Whole egg fermentation group 52.2+8.6

Preferred whole egg fermentation group 46.8±9.3

Analysis of test results: Compared with the blank control group, the whole egg fermentation group, preferred medium-dose group, obviously reduces the maze time of mice. Compared with the whole egg powder group and vinegar egg powder group, the whole egg fermentation group, preferred medium-dose group, significantly reduces the maze time of mice.

4 Sleep Improvement Experiments Fifty male Kunming mice of 18-22 g were selected and fed for one week. They were randomly divided into blank control group, whole egg powder group, vinegar egg powder group, whole egg fermentation group, preferred medium-dose group, with 10 mice in each group. The blank control group was fed with 150 mg/(kg-d) of distilled water by gavage, the whole egg powder group was fed with 150 mg/(kg-d) of the whole egg powder prepared in comparative example 5 by gavage, the vinegar egg powder group was fed with 150 mg/(kg-d) of the vinegar egg powder prepared in comparative example 6 by gavage, the whole egg fermentation group was fed with 150 mg/(kg-d) of the whole egg fermentation prepared in example 3 by gavage, and preferred medium-dose group was fed with 150 mg/(kg-d) of the whole egg fermentation prepared in example 4 by gavage. The mice were fed with by gavage once a day for 30 days.

4.1 Sub-threshold Dose Hypnosis Experiments and Sleep Latency Experiments of Barbiturate Sodium

30 min after a final intragastric administration, the sub-threshold dose of 300 mg/(kg-BW) and the injection amount of 0.1 ml/(10 gbw) were intraperitoneally injected with barbital sodium. After the subjects were injected with sub-threshold barbital sodium, the effect of barbital sodium on the sleep rate of mice was observed. 30 min after the last intragastric administration, the sub-threshold dose of 300 mg/(kg-BW) and the injection amount of 0.1 ml/(10 g-BW) were intraperitoneally injected with barbital sodium. After the subjects were injected with sub-threshold barbital sodium, the effect of barbital sodium on sleep latency of mice was observed. The results are shown in Table 9. Table 9 sleep rate founderer Experiment group Incubation period (min) Sub-threshold dose

Blank control group 21.7 34.6+3.8

Whole egg powder group 47.3 23.6±5.2

Vinegar egg powder group 50.2 21.3±5.3

Whole egg fermentation group 65.3 16.4±6.3

Preferred whole egg 66.7 66.7 15.8+4.4 fermentation group

Analysis of test results: Compared with the blank control group, the whole egg fermentation group, the optimized whole egg fermentation group, the whole egg powder group and the vinegar egg powder group can significantly improve the sleep status of the tested mice, increase the sleep rate of mice injected with sub-threshold dose of barbiturate sodium, and significantly shorten the sleep latency of mice injected with sub-threshold dose of barbiturate sodium.

4.2 Pentobarbital Sodium Induced Sleep Time Experiments in Mice 30 min after a final intragastric administration, 300 mg/(kg-d) pentobarbital sodium was injected intraperitoneally in an amount of 0.1 ml/(10 g-BW). Taking the disappearance of righting reflex as an index, the number of sleeping mice and sleep time were recorded. The results are shown in table 10. Table 10

Experiment group Sleep time (min) Sleep time extension rate (%)

Blank control group 45.7±9.2

Whole egg powder group 75.2±10.6 64.6

Vinegar egg powder group 86.1+7.2 88.4

Whole egg fermentation group 110.6+5.4 142.1

Preferred whole egg 115.5+6.2 152.8 fermentation group

Analysis of test results: Compared with the blank control group, the sleep time of the whole egg fermentation group and preferred medium-dose group was significantly prolonged with an extension rate as high as 152.8%. Compared with the whole egg powder group and vinegar egg powder group, the sleep time of mice in the whole egg fermentation group and preferred medium-dose group significantly prolonged

. 5. Blood Pressure and Lipid Regulation Experiments 5.1 ACE Inhibitory Rate Experiments Study on the activity of ACE inhibitory peptides of whole egg fermentation in vitro: Under the conditions of substrate mass concentration 40 g/l, enzymolysis pH 9.0, temperature 55°C, enzyme dose 8% (mass fraction, alkaline protease) and time 175 min, the whole egg fermentation was enzymolyzed to obtain an enzymolyzed liquid, which was then ultrafiltrated and divided into three peptide segments: < 1 kDa, 1-5 kDa and > 5 kDa, and the ACE inhibitory rates of the three component peptide segments were determined. The results are shown in Figs. 1-4. Analysis of test results: all three groups had certain ACE inhibitory activity, wherein the ACE inhibitory activity of the component < 1kDa was 84.45% at the highest, and peptide fragments with high ACE inhibitory activity existed in the < 1kDa polypeptide. The polypeptide < 1kD was selected for further separation and purification to explore the relationship between polypeptide concentration and ACE inhibitory rate. The results are shown in Fig. 5. Analysis of test results: When the concentration of polypeptide was 0.2-1.0 mg/ml, ACE inhibitory rate increased with the increase of polypeptide concentration. The higher the concentration of polypeptide, the more ACE inhibitory fragments present, and the stronger the ACE inhibitory effect was.

5.2 Blood Pressure Experiments Forty male Kunming mice of 18-22 g were selected and fed for one week. They were randomly divided into blank control group, whole egg powder group, vinegar egg powder group and whole egg fermentation group with 10 mice in each group. The blank control group was fed with 150 mg/(kg-d) of distilled water by gavage, the whole egg powder group was fed with 150 mg/(kg-d) of the whole egg powder prepared in comparative example 5 by gavage, the vinegar egg powder group was fed with 150 mg/(kg-d) of the vinegar egg powder prepared in comparative example 6 by gavage, and the whole egg fermentation group was fed with 150 mg/(kg-d) of the whole egg fermentation prepared in example 3 by gavage. The mice were fed with by gavage once a day for 28 d. The diastolic and systolic blood pressure of mice was detected and the results are shown in Figs. 6 and 7. Analysis of test results: it can be seen from Fig. 6 and 7 that the systolic and diastolic blood pressure of mice in the whole egg powder group, vinegar egg powder group and whole egg fermentation group were significantly reduced compared with the blank control group during the 28 days' gavage administration. Compared with gavage administration using the whole egg powder and vinegar egg powder of the same dose, the blood pressure of the mice in the whole egg fermentation group was obviously reduced, which indicated that the blood pressure reducing effect of the whole egg fermentation powder was obviously better than that of the whole egg powder and fermented egg powder in the process of long-term gavage administration of the whole egg fermentation. The whole egg fermentation has the function of auxiliary depressurization.

5.3 Blood Lipid Experiments Seventy male Kunming mice of 18-22 g were selected and fed for one week. They were randomly divided into blank control group, model group, whole egg powder group, vinegar egg powder group, low-dose group, medium-dose group and high-dose group with 10 mice in each group. The blank control group was fed with subcutaneous injection of 300 mg/(kg-d) normal saline every day. The model group and the administration group were fed with subcutaneous injection of D-galactose 300 mg/(kg-d) daily. The blank control group and the model group was fed with distilled water by gavage at a dose of 150 mg/(kg-d), the whole egg powder group was fed with whole egg powder prepared in comparative example 5 by gavage at a dose of 150 mg/(kg-d), the vinegar egg powder group was fed with vinegar egg powder prepared in comparative example 6 by gavage at a dose of 150 mg/(kg-d), and the high-dose group, the medium-dose group and the low-dose group was fed with the whole egg fermentation prepared in example 3 by gavage at a dose of 300 mg/(kg-d), 150 mg/(kg-d) and 20 mg/(kg-d) respectively. The preferred medium-dose group was fed with whole egg fermentation prepared in Example 4 by gavage at a dose of 150 mg/(kg-d). Each group was fed once a day for 30 continuous days. Serum TC, TG and HDL-C were detected, and the results are shown in Table 11.

Table 11

Experiment group TC(mmol/l) HDL-C(mmol/l) TG(mmol/l)

Blank control group 1.82+0.45 1.18+0.32 0.65+0.22

Model group 2.55+0.32 1.32+0.12 1.42+0.15

Whole egg powder group 2.56+0.42 1.41±0.21 0.75±0.14

Vinegar egg powder group 2.35+0.42 1.29+0.17 0.67+0.09

Low-dose group of whole egg fermentation 2.14+0.23 1.19+0.23 0.61+0.09

Medium-dose group of whole egg 2.01+0.41 1.23+0.24 0.51+0.13 fermentation

High-dose group of whole egg fermentation 1.87+0.42 1.09+0.18 0.45+0.15

Analysis of test results: Compared with the blank control group, the TC, HDL-C and TG of the model group on the 30th day were higher than those of the blank control group, indicating the success of the modeling. Compared with the blank control group, the TC, HDL-C and TG of the low-dose group, the medium-dose group and the high-dose group at the 30th day were all lower than those of the blank control group, which indicated that the whole egg fermentation has the function of reducing blood fat.

6. Life Extension Experiments 1,000 drosophilas were selected and randomly divided into blank control group, whole egg powder group, vinegar egg powder group, whole egg fermentation group, preferred medium-dose group, and 200 drosophilas each week. The blank control group was fed with basic culture medium, the whole egg powder group was fed with basic feed added with whole egg powder prepared in comparative example 5 at a dose of 50 mg/(kg-d), and the vinegar egg powder group was fed with basic feed of vinegar egg powder prepared in comparative example 6 at a dose of 50 mg/(kg-d). The whole egg fermentation group was fed with the basic feed of the whole egg fermentation prepared in example 3 at a dose of 50 mg/(kg-d). The preferred whole egg fermentation group was fed with the basic feed of the whole egg fermentation prepared in example 4 at a dose of 50 mg/(kg-d). The culture medium was changed every 3 d at a temperature of 25°C and a humidity of 65-75%. The number of dead drosophilas was recorded until all of them die and a life curve was obtained. As shown in Fig. 8, the average life, the maximum life and half of the death time were calculated, and the results are shown in table 12. Table 12

Maximum Average Life Half the time of Life extension life (Days) death rate (%)

Blank control group 66.7 36.2±1.3 36.1+0.3

Whole egg powder group 78.5 37.7+2.1 42.7+2.1 4.1

Vinegar egg powder group 76.2 38.2+2.5 42.2+2.5 5.5

Whole egg fermentation group 86.2 43.5+3.3 45.5±3.3 20.2

Preferred whole egg fermentation 88.4 45.2+2.5 46.9+2.1 24.9 group

Analysis of test results: after feeding with the same dose of whole egg powder, vinegar egg powder, whole egg fermentation and preferred whole egg fermentation, the average life span, half death time and maximum life span of drosophilas were all prolonged compared with the blank control group, and the life span extension rate of the average life span was 4.1%, 5.5%, 20.2% and 24.9% respectively. Compared with the blank control group, the average life span of drosophilas given the whole egg fermentation group, preferred medium-dose group was significantly different.

7 Regulating Function Experiments Six women with symptoms of menstrual disorder were selected, and 7.5 g of the whole egg fermentation prepared in Example 3 were taken daily for 6 months. Menstrual cycle was recorded and blood routine and hormone levels were detected. The results are shown in Tables 13 and 14. Table 13

Indicators No.0 d Six months later

Hb(g/l) 134.76+15.25 162.50+10.29

RBC(x1012 /l) 4.52+0.23 5.93+0.39

WBC(x10 9/l) 7.15+0.47 5.12±0.45

Table 14

Indicators The initial day Six months later

T(nmol/l) 2.37±0.17 3.93±0.35

E2(pmol/l) 119.48±35.67 139.07+15.67

Cort(nmol/l) 372.55±92.34 364.85+37.82

Cr(mmol/l) 109.12+31.79 138.33+21.47

Analysis of test results: the menstrual cycle of 6 women after taking whole egg fermentation returned to normal after 6 months, with an average cycle of (27.17 1.47)d and relatively neat. Hb, RBC, T, E 2 and Cr contents were significantly improved after treatment.

8. Beauty Efficacy Experiments 200 persons with chloasma were selected and randomly divided into 4 groups with 50 persons in each group, wherein three groups were whole egg powder group, vinegar egg powder group and whole egg fermentation group respectively. Whole egg powder group, vinegar egg powder group and whole egg fermentation group, each of which took 100 g/50 kg of whole egg powder, vinegar egg powder and whole egg fermentation twice a day for 60 d respectively, and the remaining group was a blank control group without taking any egg powder food. After 60 days, superoxide dismutase (SOD) and malondialdehyde (MDA) in human venous blood were detected, and the results are shown in Table 15. The effect was evaluated after 60 days by the following evaluation criteria: Observed by the naked eyes, if the chloasma area has subsided by more than 90% and the color has basically disappeared, it indicated a basically cured chloasma; observed by the naked eyes, if the chloasma area has subsided by more than 60%, and the color has obviously faded, it indicated marked effect; observed by the naked eyes, if the chloasma area has subsided by more than 30% and the color has become lighter, it indicated an improvement; the rest was invalid. The evaluation results were shown in Table 16. Table 15

Experiment group SOD(gU/l) MDA(gmol/l)

Blank control group 129.33+27.15 6.78+0.32

Whole egg powder group 141.82+32.09 4.92+0.42

Vinegar egg powder group 179.28±31.68 4.36±0.55

Whole egg fermentation group 199.18±29.52 3.72+0.67

Table 16

Basically To have a marked To be Invalid Effective cured/case effect/case improved/case /case rate/%

Whole egg powder 1 3 3 43 14 group

Vinegar egg powder 3 5 7 35 30 group

Whole egg 8 13 18 11 78 fermentation group

Analysis of test results: Compared with the blank control group, the activity of SOD in the whole egg fermentation group was significantly enhanced and the content of MOD was significantly decreased, indicating that the whole egg fermentation can improve the activity of SOD and prevent the accumulation of free radicals in the body. Compared with the whole egg powder group and the vinegar egg powder group, the whole egg fermentation group significantly improved the basic cure rate and effective rate of chloasma.

9 Experiments on Improving Osteoporosis and Effect on the Growth of Bone Metabolism and Bone Mineral Density 9.1 Effect Experiments on Growth Bone Metabolism and Bone Mineral Density Fifty male Kunming mice about 4 weeks old were selected and randomly divided into blank control group, low-dose group, medium-dose group, high-dose group and calcium feed group with 10 mice in each group. The blank control group was fed with basic feed, the high-dose group, the medium-dose group and the low-dose group were fed with the whole egg fermentation prepared in Example 3 with a dose of 300 mg/(kg-d), 150 mg/(kg-d), 20 mg/(kg-d), respectively. The calcium feed group was fed with calcium feed containing 500 mg/100 g of feed, while the blank control group was not fed with deionized water, and the other groups were fed with deionized water by gavage for 90 days. During the nourishing period, mice were weighed regularly and killed after 90 days of feeding. The right femur is stripped off to measure its length. The femur is baked to a constant weight in a 105°C oven and then weighed. Bone mineral density (BMD) of the mid-point and distal end of femur was measured by bone densitometer. The bone calcium content of the subjects was measured by atomic absorption method. The results are shown in Tables 17, 18 and 19.

Table 17

Experiment group Initial weight 28th day 56th day 90th day

Blank control group 74.2±4.5 183.8+4.8 240.8+6.2 265.7+3.5

low-dose groupofwholeegg 75.3+3.6 184.9+3.7 241.7±5.5 270.3+6.8 fermentation

Medium-dosegroupofwholeegg 74.52.7 185.8±4.9 242.2+7.2 267.2+7.3 fermentation

High-dose group of whole egg 73.2+4.9 183.5+2.5 238.5+8.6 269.4+6.7 fermentation

Calcium feed control group 76.1+3.5 185.2+4.3 245.4+6.8 271.8+4.9

Table 18

Femur length Femoral Bone calcium (L/cm) weight (m/g) content (c/mg-g1

) Blank control group 2.93+0.02 0.43+0.03 179.3+4.9

Low-dose group of whole egg fermentation 2.89+0.05 0.44+0.02 180.9±5.5

Medium-dose group of whole egg fermentation 2.94+0.06 0.48+0.03 189.8+2.6

High-dose group of whole egg fermentation 2.96+0.06 0.49+0.05 193.9+3.7

Calcium feed control group 2.92+0.03 0.49+0.02 194.8+2.9

Table 19

Bone telecentric Experiment group Femoral midpoint density density

Blank control group 0.18+0.05 0.102+0.04

Low-dose group of whole egg fermentation 0.21+0.03 0.196+0.01

Medium-dose group of whole egg fermentation 0.37+0.02 0.246+0.08

High-dose group of whole egg fermentation 0.42+0.04 0.262+0.05

Calcium feed control group 0.41+0.03 0.271±0.07

Analysis of test results: From Table 17, it can be seen that the body weight of each group of mice increased in different degrees during the Experiments, but there was no significant difference in the body weight of each group of mice during the same period, indicating that the whole egg fermentation would not affect the normal growth and development of mice. From Table 18, it can be seen that there was no significant difference in femur length and femur weight among the groups of mice. Compared with the blank control group, the bone calcium content in the medium-dose group and the high-dose group was significantly increased, indicating that the whole egg fermentation can improve the bone calcium content of mice. From Table 19, it can be seen that the density of femoral central point and bone distal end in the high-dose group, medium-dose group and low-dose group were obviously increased compared with the blank control group, which indicated that the whole egg fermentation can improve the bone density of mice.

9.2 Experiments on Improving Osteoporosis Ten people aged 50-60, 60-70 and 70-80 were selected, and pain were evaluated and recorded before taking the whole egg fermentation prepared in Example 3. Then, the whole egg fermentation was taken twice a day, 80 g each time, and pain were evaluated again 30 days later and recorded. The results are shown in Table 20. Among them, the evaluation criteria were as follows: for better improvement, pain symptoms basically disappear and BMD significantly increases; for general improvement, relief of pain symptoms and slight increase in bone mineral density; and for non improvement, no relief in pain symptoms and no increase in BMD.

Table 20

Better General Effective rate Experiment group Invalid/case improvement/case improvement/case (%)

50-60 year old group 3 3 4 60

60-70 year old group 4 3 3 70

70-80 year old group 2 4 4 60

Analysis of test results: As can be seen from Table 20, osteoporosis symptoms of all groups of people had been improved to various degrees, with the improvement rate reaching 60-70%. This indicated that the whole egg fermentation can improve joint pain, eliminate joint inflammation, supplement bone nutrition, increase bone density, and fundamentally improve senile osteoporosis.

10 Relaxing the Bowels, Regulating Constipation Experiments Sixty male Kunming mice of 18-22 g were selected and fed for one week. They were randomly divided into blank control group, model group, whole egg powder group, vinegar egg powder group, whole egg fermentation group, and preferred medium-dose group with 10 mice in each group. The blank control group and the model group were fed with common feed, the whole egg powder group was fed with common feed containing the whole egg powder prepared in comparative example 5 at a dose of 150 mg/(kg-d), the vinegar egg powder group was fed with common feed containing vinegar egg powder prepared in comparative example 6 at a dose of 150 mg/(kg-d), and the whole egg fermentation group was fed with common feed containing the whole egg fermentation prepared in example 3 at a dose of 150 mg/(kg-d), respectively. The preferred whole egg fermentation group was fed with a common feed containing the whole egg fermentation prepared in example 4 at a dose of 150 mg/(kg-d). The mice were feed for 15 days continuously to observe the small intestine propulsion Experiments and defecation Experiments of mice.

10.1 Small Intestine Propulsion Experiments At the end of the 15th day of feeding, fasting was carried out for 16 h. Five mice were taken from each group. Model group, whole egg powder group, vinegar egg powder group, whole egg fermentation group and preferred medium-dose group were fed with 5 mg/(kg-d) compound diphenoxylate vinegar solution by gavage at a dose of 20 ml/(kg-d). The blank control group was fed with the same amount of distilled water. After 30 min, the whole egg powder group, vinegar egg powder group, whole egg fermentation group and preferred medium-dose group were respectively given 10 ml/kg of ink containing corresponding whole egg powder, vinegar egg powder, whole egg fermentation and preferred whole egg fermentation at one time, mixed according to the mass ratio of 1:1. The blank control group and the model group were fed with the same amount of ink. 25 min later, the animals were killed by cervical dislocation. The abdominal cavity was immediately opened to separate the mesentery. The upper intestine from pylorus, lower end to ileocecal part was cut out, pulled into a straight line and measured without any traction. The length of intestinal canal was "total length of small intestine", and the length from pylorus to ink front was "ink propulsion length". The results are shown in Fig. 21. The ink propulsion rate was calculated according to the following formula. Ink propulsion rate (p) = Ink propulsion length / Total length of intestinal canal x100% Table 21

Experiment group Ink propulsion Total length of Ink propulsion rate length /cm intestine /cm /%

Blank control group 9.1+0.3 31.4+0.2 29.0

Model group 8.2+0.6 30.4+1.7 27.0

Whole egg powder group 10.2+1.1 30.6+1.8 33.3

Vinegar egg powder group 10.4+1.3 29.7+1.9 35.0

Whole egg fermentation group 11.9+1.1 31.1+2.8 38.3

Preferred whole egg fermentation 12.3+0.7 32.4+2.0 38.0 group

Analysis of test results: table 21 shows that compared with the blank control group, the ink propulsion distance and the ink propulsion rate of the model group were significantly lower than those of the blank control group, indicating the successful modeling. Compared with the blank control group, the ink propulsion rate of mice in the whole egg fermentation group and preferred medium-dose group was higher than that in the blank control group, which indicated that the whole egg fermentation has a function of promoting small intestine movement.

10.2 Defecation Experiments

At the end of the 15th day of feeding, fasting was carried out for 16 h. Five mice were taken from each group. The model group, whole egg powder group, vinegar egg powder group, whole egg fermentation group, and preferred medium-dose group were fed with 10 mg/(kg-d) compound diphenoxylate vinegar solution at a dose of 20 ml/kg. The blank control group was fed with the same amount of distilled water. After 30 min the whole egg powder group, vinegar egg powder group, whole egg fermentation group and preferred medium-dose group were respectively given 10 ml/kg of ink containing corresponding whole egg powder, vinegar egg powder, whole egg fermentation and preferred whole egg fermentation at one time, mixed according to the mass ratio of 1:1. Blank control group and model group were fed with the same amount of ink, and timing was started at the same time. Each mouse was raised separately and fed with feed and drinking water. The time required for each animal to excrete black stool for the first time, the number and weight of black stool excreted within 6 h were observed and recorded. The results are shown in Table 22. Table 22

Experiment group First toilet time /min Fecal grains/grains Fecal weight /g

Blank control group 157.2±1.6 23.1+2.3 0.34+1.8

Model group 228.2+4.6 17.4+6.7 0.22+0.13

Whole egg powder group 210.2+61.1 19.6+5.8 0.24+0.14

Vinegar egg powder group 170.4+39.3 22.7+7.9 0.35+0.22

Whole egg fermentation group 146.9+53.1 24.1+6.8 0.38+0.14

Preferred whole egg fermentation 144.5+23.6 24.3+4.7 0.39+0.09 group

Analysis of test results: table 22 shows that compared with the blank control group, the time for mice in the model group to discharge the first black stool was significantly longer than that in the blank control group, and the number of stool particles and stool density were significantly lower than that in the blank control group, indicating the success of constipation model. Compared with the model control group, in the whole egg fermentation group and preferred medium-dose group, the first defecation time is obviously shortened and the number and weight of feces is significantly increased, which indicated that the whole egg fermentation has the function of promoting defecation.

The number of devices and processing scale described herein are used to simplify the description of the present invention. Applications, modifications and variations of the present invention will be apparent to those skilled in the art. Although the embodiment of the present invention has been disclosed above, it is not limited to the application listed in the specification and the embodiments, it can be fully applied to various fields suitable for the present invention, and additional modifications can be easily implemented by those skilled in the art. Therefore, the present invention is not limited to the specific details and the examples shown and described herein without departing from the general concept defined by the claims and the equivalent scope.

Claims (10)

1. What is claimed is: 1. A method for producing high bioactivity whole egg fermentation, comprising: step 1 in which after poultry eggs or chicken embryo eggs are cleaned, sterilized and aired, edible organic acid is added therein by 1-3 times the weight of the poultry eggs to submerged and soaked the eggs for 1-6 days until eggshells are completely dissolved; the acid solution is filtered and collected, and the egg membranes are rinsed with water to remove insoluble substances and pigment from the eggshells, and then are submerged and soaked again in the collected acid solution; step 2 in which the egg is stirred to release the egg membranes and egg liquid, which are subjected to a continuous hydrolysis for 1-24 months under ultrasonic waves until the whole egg liquid is homogenized and free of lumps, then lipase or phospholipase with the mass of 1/1000-1/100 times of the eggs and protease with the mass of 1/500-1/50 times of the eggs are added therein to continue enzymatic hydrolysis for 1-5 hours to obtain hydrolyzed whole egg liquid; step 3 in which the hydrolyzed whole egg liquid is mixed with brown sugar and inulin prebiotics, then is homogenized, sterilized, cooled, inoculated with yeast and lactic acid bacteria, wherein the inoculation amount of the bacterial liquid is 1-5% of the volume of the hydrolyzed whole egg liquid, and finally is fermented at 25-45°C environment for 1-15 days to obtain whole egg fermentation liquid; and step 4 in which the whole egg fermentation liquid is filtered, followed by adding therein with cyclodextrin under ultrasonic waves, and then is dispersed at high speed, mixed for homogenization, embedded, cooled at a low temperature environment of 0-5°C and stood, and finally centrifuged to remove cholesterol insoluble substances to obtain separation liquid which is then freeze-dried to obtain whole egg fermentation.
2. 2. The method of claim 1, wherein the edible organic acid is selected from the group consisting of lactic acid, malic acid, butyric acid, acetic acid, edible rice vinegar, fruit vinegar and/or citric acid, and the concentration of the edible organic acid is 5-50wt%.
3. 3. The method of claim 1, wherein the lipase is derived from Aspergillus oryzae or Rhizopus oryzae, the phospholipase is selected from phospholipase Al, A2, C and/or D, and the protease is selected from bromelain, papain, ficin, pepsin, trypsin and/or acidic protease or neutral protease or alkaline protease derived from Aspergillus niger and Bacillus.
4. 4. The method of claim 1, wherein the yeast is Saccharomyces cerevisiae, and the lactobacillus is Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus reuteri, Lactobacillus acidophilus and/or Propionibacterium fischeri.
5. 5. The method of claim 1, wherein the edible organic acid in the first step is subjected to a freezing-thawing pretreatment comprising placing the edible organic acid in -20°C environment with an ice water mixture at 0°C with 0.5 times the weight of the edible organic acid being added therein to form ice crystals through dissolving and stirring, adding another ice water mixture at 0°C with 0.5 times the weight of the edible organic acid to form ice crystals through dissolving and stirring, and placing the mixture obtained thereby in -20°C environment to freeze for 1 h and then mashing the same to fragments which are heated to 0°C at a rate of1C/min, further adding therein with ice water mixture at 0°C to make the concentration of the edible organic acid reaches 20wt%, and then conducting refrigeration for 1 h.
6. 6. Whole egg fermentation, prepared by the production method according to any one of claims 1 to 5, the content of organic calcium of which is not less than 2wt%, and the content of polypeptide protein of which is not less than 20wt%, wherein the molecular weight distribution of polypeptide protein is concentrated between 0.5 and 3k, and the content ratio of small molecule polypeptide with molecular weight distribution concentrated below 1k is not less than 60wt%.
7. 7. Foods containing whole egg fermentation, comprising the whole egg fermentation of claim 6 and food ingredients or additives, which are used as nutritional functional foods, special foods and special medical foods, health foods and non-medical uses in the food field.
8. 8. The application of the whole egg fermentation of claim 6 in preparing products for preventing hypertension and hyperlipidemia.
9. 9. The application of the whole egg fermentation of claim 6 in preparing products for increasing bone density, improving osteoporosis, enhancing physical strength, enhancing immunity, prolonging life and resisting aging.
10. 10. The application of the whole egg fermentation of claim 6 in preparing products for regulating menstruation, caring skin, moistening intestine, regulating constipation, improving intelligence and improving sleep.

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