CN102160563B - Preparation method of anti-fatigue oligopeptides biscuits - Google Patents

Abstract

The invention discloses a preparation method of anti-fatigue oligopeptide biscuit, which comprises three parts: preparation of casein oligopeptide powder, preparation of wolfberry juice powder and preparation of anti-fatigue casein oligopeptide biscuit. After enzymatically hydrolyzing the prepared cow's milk casein and goat's milk casein with neutral protease and alkaline protease under specified conditions, most of the casein becomes oligopeptides that are easy to absorb and utilize. Oligopeptide powder and wolfberry juice powder can not only increase the nutritional value of biscuits, but also enhance the antioxidant activity and anti-fatigue function of biscuits. The anti-fatigue oligopeptide biscuit prepared by the method of the invention is a good food for tourism, disaster relief, army, workers' missed meals, and students' meals during breaks, and has extremely broad market prospects.

Description

A kind of preparation method of anti-fatigue oligopeptide biscuit

technical field

The invention belongs to the field of food processing, and relates to a health food and its preparation, in particular to a preparation method of an anti-fatigue oligopeptide biscuit.

Background technique

Fatigue is a complex and comprehensive physiological phenomenon. Fatigue is the inability of the body's physiological processes to sustain its function at a specified level and/or maintain a predetermined exercise intensity. The phenomenon that the body's exercise capacity is temporarily reduced due to physiological and biochemical changes caused by exercise is called exercise-induced fatigue. Sports fatigue can be divided into physical fatigue and psychological fatigue. Physical fatigue is mainly manifested as a decrease in exercise capacity, and psychological fatigue is mainly manifested as a change in behavior. Fatigue involves many physiological and biochemical factors. It is a normal physiological phenomenon that must appear when the human body's mental or physical activities reach a certain stage. a harbinger.

Fatigue is a common problem in everyday life. Zhan Fashang (2006) reviewed and reported that the fatigue incidence rate of middle school students was 19.6% to 71.0%, that of private enterprise employees was 51.6%, and that of foreign countries was also between 7% and 45%. According to the survey of the World Health Organization, about 35% of the people in the world are in a state of fatigue, especially the middle-aged male population is as high as 60-75%. With the rapid dissemination of knowledge, the accelerated pace of work and life, the competition in all stages of life is becoming increasingly fierce. However, human physiological functions have not evolved accordingly. Such an overloaded operation will inevitably lead to the reduction of athletic ability and work efficiency, and the increase of accidents. If fatigue is not recovered in time and gradually accumulates after the occurrence of fatigue, it will also lead to "overwork" and "chronic fatigue syndrome", which will cause endocrine disorders, decreased immunity, and even organic diseases in the body. serious health hazard.

At present, various measures for eliminating fatigue have their own unsatisfactory effects. For example, vitamins and amino acid preparations are safe and effective, but expensive, and most anti-fatigue preparations often contain stimulant ingredients that are harmful to the human body. Because medicine and its composition can produce side effects to the body, there is a "three-point poison in medicine" in people's psychology, which makes most people stay away from it. According to the data retrieval conducted by the applicant, up to now, there is no report on the patent or literature related to the oligopeptide biscuit with anti-fatigue function.

Active peptides have extremely strong efficacy diversity. The active peptides that have been discovered have many functions such as lowering blood pressure, lowering blood sugar, lowering fat and weight loss, immunity, anti-oxidation, anti-thrombosis, promoting digestion, promoting mineral absorption and nerve regulation, etc. A variety of peptide health foods have been successfully developed. After the protein is acted upon by enzymes, it can not only be absorbed and utilized in the form of free amino acids, but is more digested and absorbed in the form of oligopeptides, and the absorption of oligopeptides has the advantages of fast transport, low energy consumption, not easy to saturate, and no absorption characteristics of competition. Oligopeptides are easier to digest and absorb than free amino acids. Cattle and goat milk casein is the most important protein in bovine and goat milk. Under certain conditions, the casein can be hydrolyzed into oligopeptides composed of 2-10 amino acid residues with appropriate protease. Japan, France, Denmark, Taiwan and China all have peptide foods and additives on the market. Biscuit is an important baked food that the public likes to eat. It has a long shelf life, is convenient to eat, and has a variety of varieties. It is suitable for different groups of people and different occasions. However, there is a problem of single nutrition, and functional biscuits that are urgently needed in the market are very scarce. For example, a Chinese patent application discloses a polypeptide biscuit (patent number: ZL 01115245.1). This patent adds casein compound polypeptides to biscuit raw materials to make polypeptide biscuits, making it "have a special therapeutic effect on certain diseases". But there is no mention of oligopeptide biscuits with anti-fatigue effect.

Contents of the invention

In view of the prevalence of fatigue and the defects or deficiencies in biscuits, the purpose of the present invention is to provide an anti-fatigue oligopeptide biscuit and its preparation method. Enzymatic hydrolysis of milk casein, the enzymatic hydrolyzate is roughly separated to remove enzymes and macromolecular proteins, desalted with macroporous non-polar resin, and spray-dried to obtain casein oligopeptide powder. Casein oligopeptide powder and wolfberry juice powder are added to the biscuit base material, and the rest are processed into the oligopeptide biscuit with anti-fatigue effect according to the conventional processing method of the biscuit.

In order to realize above-mentioned task, the present invention adopts following technical solution:

A preparation method of anti-fatigue oligopeptide biscuits is characterized in that it comprises the following steps:

1) Preparation of casein oligopeptide powder

Take fresh cow's milk or goat's milk, filter to remove impurities, centrifuge at low temperature to skim, use 2 mol/L HCl solution to adjust the pH value of skim goat milk to 4.3, or use 2 mol/L HCl solution to adjust the pH value of skim milk to 4.6, stand still 30min, 3000 r/min, centrifuge for 10min, discard the supernatant, wash the precipitate twice with acetic acid-sodium acetate buffer solution with pH 4.3 and 4.6 respectively, centrifuge at 3000 r/min for 10 min after washing, and the obtained precipitate is Crude casein.

Weigh a certain amount of crude casein and place it in an enzymolysis tank, dissolve it with 0.1 mol/L sodium hydroxide solution, and distill the volume to the specified volume with distilled water to obtain a 60 g/kg cow's milk casein solution or goat's casein solution Solution, adjust the pH value of cow's milk casein solution or goat's casein solution to 7.0-8.0, heat at 95°C for 5 minutes, cool to 45°C-55°C, add 3000 U/g-5000 U/g of neutral protease and 200 U/g～300 U/g alkaline protease is hydrolyzed at constant temperature, and 2 mol/L NaOH solution is added during the hydrolysis to keep it within the required pH value range; after 24 hours of enzymolysis, the enzyme is extinguished at 95°C for 10 minutes, cooled , centrifuge at 10000 r/min for 15 min to remove enzymes, unenzymolyzed proteins and other macromolecular substances, desalt with non-polar macroporous resin, add excipients, concentrate in a conventional vacuum, and spray dry to obtain casein oligopeptides pink.

2) Preparation of wolfberry juice powder

Screen high-quality dried goji berries, wash away the sediment attached to the goji berries with running water, and drain the water. Mix wolfberry and purified water at a mass ratio of 1:3 to 1:10, add citric acid and vitamin C to protect the color, soak at 50-80°C for 10-30 minutes, break and beat after soaking, and then heat at 70°C-75°C Under the conditions of dynamic leaching for 1 hour, the rotation speed is 50-100 r/min, after the leaching is completed, goji berry juice is obtained by pressure filtration.

According to the weight of the filter residue, add 3 to 10 times the amount of pure water and color protection solution to the filter residue and extract once more according to the above method. The wolfberry juice obtained from the two extractions is mixed, finely filtered, added auxiliary materials, concentrated in a conventional vacuum, and spray-dried to obtain the wolfberry juice powder.

3) Preparation of anti-fatigue casein oligopeptide biscuits

In the known biscuit powder adjustment process, the casein oligopeptide powder and wolfberry juice powder are evenly mixed with flour and auxiliary materials, and processed into anti-fatigue oligopeptide biscuits according to the known biscuit processing technology. Among them, casein oligopeptide powder accounts for 0.3%-5.2% of the total mass of biscuit ingredients, and wolfberry juice powder accounts for 0.05-0.60% of the total mass of biscuit ingredients.

The preferred ingredients in terms of weight percentage are: casein oligopeptide powder accounts for 2.0%-3.2% of the total mass of biscuit ingredients, wolfberry juice powder accounts for 0.2%-0.3% of the total mass of biscuit ingredients.

The casein oligopeptide powder can also use one or more combinations of proteins such as whey protein, egg protein, hemoglobin, collagen, livestock and poultry meat, fish, shrimp and shellfish meat under suitable protease and corresponding conditions. Hydrolyzed oligopeptide powder instead.

The wolfberry juice powder can also be directly processed from fresh wolfberry without soaking.

The wolfberry juice powder can also be replaced by one or more combinations of mulberry juice powder, Jiaogulan juice powder and tea juice powder.

The mulberry juice powder, Gynostemma pentaphyllum juice powder and tea juice powder can be obtained by modifying the method of the present invention or by extracting, spray-drying or freeze-drying by other known methods.

The anti-fatigue oligopeptide biscuits prepared by the present invention bring the following technical effects:

(1) After enzymatically hydrolyzing bovine and goat milk casein with neutral protease and alkaline protease under specified conditions, most of the hydrolyzate becomes oligopeptides that are easy to absorb and utilize.

(2) Oligopeptide powder and wolfberry juice powder can not only increase the nutritional value of biscuits, but also enhance the antioxidant activity of biscuits.

(3) The prepared anti-fatigue oligopeptide biscuits have obvious anti-fatigue effect on the body.

(4) The anti-fatigue oligopeptide biscuit prepared by the method provided by the present invention is a good food for tourism, disaster relief, army, workers' missed meals, and students' lunches between classes, and has a very broad market prospect.

Description of drawings

Figure 1 is the gel chromatogram (254 nm) of goat cheese casein hydrolyzate.

Below in conjunction with the embodiment given by the inventor, the present invention will be further described in detail.

Detailed ways

The preparation method of the anti-fatigue oligopeptide biscuit of the present invention comprises three parts: preparation of casein oligopeptide powder, preparation of wolfberry juice powder and preparation of anti-fatigue casein oligopeptide biscuit.

The preparation of casein oligopeptide powder is to hydrolyze cow milk or goat milk casein with neutral protease and alkaline protease for a specified time under the conditions of specified pH, temperature and enzyme amount, then inactivate the enzyme, cool, and centrifuge to remove the enzyme and unused Enzymatically hydrolyzed protein macromolecules are spray-dried.

The wolfberry juice powder is prepared by screening, cleaning, color-protecting, beating, leaching, press-filtering, fine-filtering, and spray-drying dry wolfberry that is free from diseases, insect pests, and mildew.

The preparation of anti-fatigue casein oligopeptide biscuits is to mix the prepared casein oligopeptide powder and wolfberry juice powder with flour and other auxiliary materials in the known biscuit mixing process, and process them into anti-fatigue casein oligopeptides according to the known biscuit processing technology biscuit. Among them, casein oligopeptide powder accounts for 2.0% to 3.2% of the total mass of biscuit ingredients, and wolfberry juice powder accounts for 0.2% to 0.3% of the total mass of biscuit ingredients.

The casein oligopeptide powder can also use one or more combinations of proteins such as whey protein, egg protein, hemoglobin, collagen, livestock and poultry meat, fish, shrimp and shellfish meat under suitable protease and corresponding conditions. Hydrolyzed oligopeptide powder instead.

The wolfberry juice powder can also be replaced by one or more combinations of mulberry juice powder, Jiaogulan juice powder and tea juice powder.

The mulberry juice powder, Jiaogulan juice powder and tea juice powder can be obtained by extracting, spray-drying or freeze-drying according to the preparation method of wolfberry juice powder of the present invention or other known methods.

The main raw material of known biscuit is flour, and auxiliary material comprises egg, sugar, oil, salt, baking soda, ammonium bicarbonate etc. Its processing technology has been very mature, including preparing premixed pulp, mixing dough, rolling, printing and molding, baking, cooling, packaging and so on.

The following are specific examples given by the inventors, but the present invention is not limited to these examples. In the present invention, the following references are cited.

【1】Zhan Fashang. Status quo of fatigue research[J]. Chinese Behavioral Medicine Science, 2006, 15(2): 188-189.

【2】Chen Yuanyuan, Shi Hui, Le Guowei. Research on the relationship between antioxidant activity and anti-fatigue of soybean oligopeptides with different molecular weights[J]. Amino Acids and Biological Resources, 2008, 30(2): 59-62.

【3】Gu Fang, Qin Yide, Dong Huasheng, etc. Study on the Antioxidative and Antifatigue Effects of Milk-derived Immunomodulatory Peptides in Mice[J]. Journal of Nutrition, 2006, 28(4):326-328.

【4】Huang Yusan. Implementation Manual of Health Food Inspection and Evaluation Technical Specifications [M]. Beijing: Tsinghua Tongfang Electronic Publishing House, 2003, 756-701.

Example 1:

(1) Take fresh goat milk, filter it to remove impurities, and centrifuge it at low temperature to degrease. Use 2 mol/L HCl solution to adjust the pH value to 4.3, stand still for 30 min, centrifuge at 3000 r/min for 10 min, discard the supernatant, wash the precipitate twice with acetic acid-sodium acetate buffer solution with pH 4.3, after washing Centrifuge at 3000 r/min for 10 minutes, and the obtained precipitate is crude casein;

A certain amount of crude casein was weighed and placed in an enzymatic hydrolysis tank, dissolved with 0.1 mol/L sodium hydroxide solution, and distilled water was used to set the volume to the specified volume to obtain a 60 g/kg goat milk casein solution.

( 2) Prepare a 60 g/kg goat milk casein solution, adjust the pH to 7.5, heat at 95°C for 5 minutes, cool to 50°C, add 3500 U/g of neutral protease and 250 U/g of alkaline protease at 50°C Constant temperature hydrolysis, add 2 mol/L NaOH solution during hydrolysis to keep it within the required pH range. After 24 hours of enzymatic hydrolysis, inactivate the enzyme at 95°C for 10 minutes, cool, and centrifuge at 10,000 r/min for 15 minutes to remove macromolecular substances such as enzymes and unenzymolyzed proteins. After desalting with non-polar macroporous resins such as LS106, add auxiliary materials, vacuum Concentrate and spray dry to obtain casein oligopeptide powder.

(3) Select high-quality dry wolfberry without pests and mildew, wash it, mix dry wolfberry with purified water at a mass ratio of 1:5, add 1.2 g/kg citric acid and 0.5 g/kg vitamin C, 70 Soak at ℃ for 20 minutes, crush and beat, and then extract dynamically at 70-75℃ for 1 hour at a speed of 50-100 r/min. After the extraction is completed, press filter to obtain wolfberry juice. Add 5 times the amount of purified water and color-protecting agent to the filter residue and extract it once more in the same way. The wolfberry juice obtained from the two extractions is mixed, finely filtered, added auxiliary materials, concentrated in a conventional vacuum, and spray-dried to obtain the wolfberry juice powder.

(4) In the conventional biscuit powder mixing process, the casein oligopeptide powder and wolfberry juice powder are evenly mixed with flour and other auxiliary materials, and processed into anti-fatigue oligopeptide biscuits according to the known biscuit processing technology. Among them, goat milk casein oligopeptide powder accounts for 2.6% of the total mass of biscuit ingredients, and wolfberry juice powder accounts for 0.26% of the total mass of biscuit ingredients.

Example 2:

(1) Fresh milk is filtered to remove impurities, and centrifuged at low temperature to degrease. Use 2 mol/L HCl solution to adjust the pH value to 4.6, stand still for 30 min, centrifuge at 3000 r/min for 10 min, discard the supernatant, wash the precipitate twice with acetic acid-sodium acetate buffer solution with pH 4.6, after washing Centrifuge at 3000 r/min for 10 min, and the obtained precipitate is crude casein;

Weigh a certain amount of crude casein and place it in an enzymolysis tank, dissolve it with 0.1 mol/L sodium hydroxide solution, and distill the volume to the specified volume with distilled water to obtain a 60 g/kg milk casein solution.

( 2) Prepare a 60 g/kg milk casein solution, adjust the pH to 7.5, heat at 95°C for 5 min, cool to 50°C, add 4000 U/g neutral protease and 200 U/g alkaline protease to keep the temperature For hydrolysis, add 2 mol/L NaOH solution during the hydrolysis process to keep it within the required pH range. After 24 hours of enzymatic hydrolysis, inactivate the enzyme at 95°C for 10 minutes, cool, and centrifuge at 10,000 r/min for 15 minutes to remove macromolecular substances such as enzymes and unenzymolyzed proteins. After desalting with non-polar macroporous resins such as LS106, add auxiliary materials, vacuum Concentrate and spray dry to obtain casein oligopeptide powder.

(3) The preparation of wolfberry juice powder is the same as in Example 1.

(4) In the known biscuit powder mixing process, the casein oligopeptide powder and wolfberry juice powder are evenly mixed with flour and other auxiliary materials, and processed into anti-fatigue oligopeptide biscuits according to the known biscuit processing technology. Among them, cow milk casein oligopeptide powder accounts for 2.4% of the total mass of biscuit ingredients, and wolfberry juice powder accounts for 0.28% of the total mass of biscuit ingredients.

Example 3:

The difference between this embodiment and Example 1 is that the enzyme amounts of neutral protease and alkaline protease reenzyme are 4000 U/g and 250 U/g respectively. All the other are the same as embodiment 1, can reach the purpose of embodiment 1 equally.

Example 4:

The difference between this embodiment and Example 2 is that the enzyme amounts of neutral protease and alkaline protease reenzyme are 4500 U/g and 200 U/g respectively. In the biscuit powder mixing process, replace wolfberry juice powder with mulberry juice powder, which accounts for 0.24% of the total mass of biscuit ingredients, and the rest are the same as in Example 2, and the purpose of Example 2 can be achieved equally.

Example 5:

The difference between this embodiment and Example 1 is that 0.24% mulberry juice powder is used instead of Lycium barbarum juice powder in the biscuit powder adjustment process, and all the other are the same as Example 1, and the purpose of Example 1 can be achieved equally.

Embodiment 6:

The difference between this embodiment and embodiment 1 is that 0.22% tea juice powder is used to replace wolfberry juice powder in the biscuit powder adjustment process, and all the other are the same as embodiment 1, and the purpose of embodiment 1 can be achieved equally.

Embodiment 7:

The difference between this embodiment and Example 2 is that 0.23% Gynostemma juice powder is used to replace Lycium barbarum juice powder in the biscuit powder adjustment process, all the other are the same as Example 2, and the purpose of Example 2 can be achieved equally.

Embodiment 8:

The difference between this example and Example 2 is that the wolfberry selected is fresh wolfberry, 1.2 g/kg citric acid and 0.5 g/kg vitamin C are directly added, soaked at 70 ° C for 15 min, crushed and beaten, and then heated at 70-75 ° C Under the conditions of dynamic leaching for 1 hour, the rotation speed is 50-100 r/min, after the leaching is completed, goji berry juice is obtained by pressure filtration. Add 5 times the amount of purified water and color-protecting agent to the filter residue and extract it once more in the same way. The wolfberry juice obtained from the two extractions is mixed, finely filtered, added auxiliary materials, concentrated in a conventional vacuum, and spray-dried to obtain the wolfberry juice powder. All the other are the same as embodiment 2, can reach the purpose of embodiment 2 equally.

The neutral protease and alkaline protease reenzyme enzymatic hydrolysis of bovine and goat milk casein used in the above-mentioned examples are formulated on the basis of multiple scientific experiments by the inventor, as long as they are within the scope of the technical scheme of the application, they can be Reach the object of the present invention. The proteases and hydrolysis conditions used for other protein hydrolysis can be obtained through experiments by those skilled in the art.

According to the technical solution provided by the present invention, in addition to making anti-fatigue oligopeptide biscuits, other carriers can also be added to make anti-fatigue food in various forms such as liquid, granule, and powder.

The anti-fatigue oligopeptide biscuit has a simple preparation process and is easy for large-scale production. It is a good food for tourism, disaster relief, army, workers' missed meals, and students' inter-class supplementary meals, and has great development prospects.

Precautions:

Because wolfberry juice powder has a strong effect of warming the body, people who suffer from high blood pressure, have a cold and fever, have inflammation in the body (such as oral ulcers), or have diarrhea should eat less or stop eating temporarily. Long-term consumption of anti-fatigue oligopeptide biscuits added with wolfberry juice powder in large quantities can increase libido. It is recommended that teenagers with good liver and kidney functions do not consume large quantities for a long time.

The following are the goat milk casein enzymatic hydrolyzate separation test, anti-oxidation test and anti-fatigue function test according to the new method in the 2004 Ministry of Health "Notice on Issuing the Revised Items of "Health Food Functional Evaluation Procedures and Test Methods"" .

Test 1: Gel chromatography separation test of goat milk casein hydrolyzate

Goat milk casein was enzymatically hydrolyzed and treated with neutral protease and alkaline protease multiple enzymes under specified conditions, and the hydrolyzate was separated with a gel chromatography peptide column (Figure 1). Compared with the standard molecular weight, the goat milk casein hydrolyzate was separated into six relatively independent peaks from peak 1 to peak 6, and their relative content and molecular weight distribution are shown in Table 1. The gel-separated components in the goat cheese casein hydrolyzate were mainly concentrated In peak 2, peak 4 and peak 5, its content accounts for more than 83% of the total hydrolyzate, and they are oligopeptides composed of about 2-10 amino acids.

Table 1 Relative content and molecular weight distribution of gel-separated peaks of goat cheese casein hydrolyzate

Note 1: Because the content of peak 3 is too small, peak 3 and peak 4 are combined and called peak 4.

Note 2: Peak 6 is free amino acid.

Test 2: Anti-oxidation function test of anti-fatigue oligopeptide biscuits

The antioxidant activity of goat milk casein was significantly enhanced after neutral protease and alkaline protease enzymatic hydrolysis (Table 2), and the EC ₅₀ value of HO scavenging rate (the concentration of the sample when the free radical scavenging rate reached 50%) increased to 3.59 times of the original, the scavenging capacity of ABTS ⁺ increased to 158.72 times of the original, and the scavenging capacity of O ₂ ^- and DPPH increased from undetectable to 223.77 μ g/mL and 244.27 μ g/mL respectively. very obvious. Chen Yuanyuan et al. (2008) showed that there was a significant positive correlation between antioxidant activity and anti-fatigue function. Combined with the test results of Gu Fang et al. (2006), the oligopeptides of bovine and goat milk casein hydrolyzate with strong antioxidant activity also have Strong anti-fatigue function.

Table 2 EC of antioxidant activity of goat milk casein and its oligopeptides ₅₀ μg/mL

Note: "-" means not detected.

Test 3: Anti-fatigue function test of anti-fatigue oligopeptide biscuits

120 male Kunming mice of clean grade, with an average body weight of 23±2 g, were randomly divided into 4 groups after adaptive feeding for 3 days, namely low, medium and high dose groups and a control group, with 30 mice in each group. The biscuits in the low, medium and high dose groups all contained 0.26 g/100g of wolfberry juice powder, and 1.3 g/100g, 2.6 g/100g and 3.9 g/100g of goat milk casein oligopeptide powder respectively. The biscuit prepared by the present invention is 16.7 g/kg (equivalent to 10 times of the daily biscuit 100 g/60kg body weight of an adult), and the control group is fed with ordinary biscuit (without adding goat casein oligopeptide powder and wolfberry juice powder) 16.7 g/kg, once a day. During the rest of the time, they were free to drink water and eat standard diets for 2 weeks. 12 hours after the last feeding of biscuits, 10 mice were randomly selected from each group, and a lead wire of 5% of their body weight was loaded on the base of the tail of the mice, and placed in a swimming tank with a water depth of 40 cm and a water temperature of 25°C±0.5°C. Record the swimming time from the start of swimming to the time when the mouse’s head is completely immersed in the water for 8 seconds and cannot emerge from the water; 10 mice are randomly selected from each group to measure the content of liver glycogen and muscle glycogen according to Huang Yusan’s method; Blood urea nitrogen content was measured in 10 mice.

3.1 Effect of anti-fatigue oligopeptide biscuits on weight gain of mice

There were no abnormalities in the mental state, diet, hair state, feces and other conditions of the mice during the test. The weight of the mice at the end of the test was significantly higher than that at the beginning of the test, but there was no significant difference in the weight of the mice between the groups before and after the test (P>0.05) (Table 3). Anti-fatigue oligopeptide biscuits had no significant effect on the weight of the mice among the test groups.

±s，n=10) Table 3: Effects of anti-fatigue oligopeptide biscuits on body weight of mice (

±s, n=10)

Note: There is no significant difference between the symbols with the same lowercase letters P>0.05.

3.2 Effects of anti-fatigue oligopeptide biscuits on exercise endurance, liver glycogen, muscle glycogen and blood urea nitrogen content of mice

Compared with the control group, the swimming time, muscle glycogen and liver glycogen content and blood urea nitrogen content of the anti-fatigue oligopeptide biscuit groups were significantly or extremely significantly different (P<0.05, P<0.01 ) (Table 4), indicating that anti-fatigue oligopeptide biscuits can significantly prolong the weight-bearing swimming time of mice, prolong the content of liver glycogen and muscle glycogen, reduce the content of serum urea nitrogen, and have a significant anti-fatigue effect, among which The effect of the dose group is the best, while the anti-fatigue effect of high doses tends to decrease, indicating that the anti-fatigue oligopeptide biscuit as a functional food should not be taken in excess, otherwise it will cause certain harm to the body's health.

±s，n=10) Table 4: Effects of anti-fatigue oligopeptide biscuits on mouse exercise endurance, liver glycogen muscle glycogen and blood urea nitrogen content (

±s, n=10)

Note: Different lowercase letters a and b mean significant difference P<0.05; different uppercase letters A and B mean extremely significant difference P<0.01.

Claims (7)

1. a preparation method of anti-fatigue oligopeptide biscuit, is characterized in that, prepares according to the following steps: 1) Preparation of casein oligopeptide powder Take fresh cow's milk or goat's milk, filter and remove impurities, centrifuge and degrease at low temperature, adjust the pH value of skim goat milk to 4.3 with 2 mol/L HCl solution, or adjust the pH value of skim milk to 4.6 with 2 mol/L HCl solution; stand still for 30 min, 3000 r/min, centrifuge for 10 min, discard the supernatant, wash the precipitate twice with acetic acid-sodium acetate buffer solution with pH 4.3 and pH 4.6 respectively, centrifuge at 3000 r/min for 10 min after washing, and obtain the precipitate The product is crude casein; Weigh a certain amount of crude casein and place it in an enzymolysis tank, use 0.1 mol/L sodium hydroxide solution to help dissolve it, and distill the volume to the specified volume with distilled water to obtain a 60 g/kg cow's milk or goat's casein solution, adjust pH to 7.0-8.0, heat at 95°C for 5 minutes, cool to 45°C-55°C, add 3000 U/g-5000 U/g neutral protease and 200 U/g-300 U/g alkaline protease to hydrolyze at constant temperature During the hydrolysis process, 2 mol/L NaOH solution was added to keep it within the required pH range; after 24 hours of enzymolysis, the enzyme was inactivated at 95°C for 10 minutes, cooled, and centrifuged at 10,000 r/min for 15 minutes to remove the enzyme and Unenzymolyzed protein and other macromolecular substances are desalted with non-polar macroporous resin, then add excipients, concentrate in a conventional vacuum, and spray dry to obtain casein oligopeptide powder; 2) Preparation of wolfberry juice powder Screen high-quality dried goji berries, wash away the sediment attached to goji berries with flowing water, drain the water, mix dried goji berries with purified water at a mass ratio of 1:3 to 1:10, add citric acid and vitamin C to protect the color, Soak at 50℃～80℃ for 10min～30min, crush and beat after soaking, then dynamically leaching at 70℃～75℃ for 1h, rotating speed 50r/min～100r/min, after leaching, press filter get goji berry juice; According to the weight of the filter residue, add 3 to 10 times of pure water and color protection solution and extract once more according to the above method; Mix the wolfberry juice obtained from the two extractions, fine filter, add auxiliary materials, concentrate in vacuum according to conventional methods, and spray dry to obtain wolfberry juice powder; 3) Preparation of anti-fatigue casein oligopeptide biscuits In the conventional biscuit mixing process, the casein oligopeptide powder and wolfberry juice powder are mixed evenly with flour and auxiliary materials, and processed into anti-fatigue oligopeptide biscuits according to the known biscuit processing technology; the casein oligopeptide powder accounts for the total mass of biscuit ingredients 0.3% to 5.2% of the total mass of biscuit ingredients, and wolfberry juice powder accounts for 0.05 to 0.60% of the total mass of biscuit ingredients. 2. The method according to claim 1, wherein the casein oligopeptide powder accounts for 2.0%-3.2% of the total mass of biscuit ingredients, and wolfberry juice powder accounts for 0.2%-0.3% of the total mass of biscuit ingredients. 3. The method as claimed in claim 1, characterized in that, the casein oligopeptide powder or one of whey protein, egg protein, hemoglobin, collagen, livestock and poultry meat, fish, shrimp and shellfish protein Or more than one combination can be replaced by oligopeptide powder hydrolyzed under appropriate protease and corresponding conditions. 4. The method according to claim 1, wherein the wolfberry juice powder is obtained by crushing and beating fresh wolfberry, extracting, vacuum concentrating, and spray drying. 5. The method according to claim 4, wherein the wolfberry juice powder is replaced by one or more combinations of mulberry juice powder, Gynostemma pentaphyllum juice powder and tea juice powder. 6. The method according to claim 5, wherein the mulberry juice powder, Jiaogulan juice powder and tea juice powder are extracted, spray-dried or freeze-dried according to the preparation method of wolfberry juice powder in step 2) or other known methods. become. 7. The anti-fatigue casein oligopeptide biscuits prepared by the method according to one of claims 1 to 6.

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