CN109222110A - A kind of antifatigue soluble granule and preparation method of the polypeptide dry powder containing Goat Placenta - Google Patents

A kind of antifatigue soluble granule and preparation method of the polypeptide dry powder containing Goat Placenta Download PDF

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CN109222110A
CN109222110A CN201810785568.7A CN201810785568A CN109222110A CN 109222110 A CN109222110 A CN 109222110A CN 201810785568 A CN201810785568 A CN 201810785568A CN 109222110 A CN109222110 A CN 109222110A
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goat placenta
polypeptide
dry powder
preparation
protease
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闵红岗
宋新波
边育红
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17—Amino acids, peptides or proteins
    • A23L33/18—Peptides; Protein hydrolysates
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
    • A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
    • A23L5/25—Removal of unwanted matter, e.g. deodorisation or detoxification using enzymes

Abstract

It is using fresh Goat Placenta as raw material, preprocessed, homogenate, enzymatic hydrolysis, the de- bitter, centrifugation of deodorant, concentration, spray drying prepare Goat Placenta polypeptide powder the invention discloses a kind of preparation method of Goat Placenta polypeptide soluble particle；Goat Placenta polypeptide powder and synanthrin are mixed in proportion, granule is then made according to a conventional method, dispensed, sterilized to get finished product.Granule of the invention is full of nutrition, be easy to absorb, flavor taste is good and it is antifatigue to have the function of, adjusts immune, beautifying face and moistering lotion.Preparation method rational technology, production is simple, strong operability, is suitble to further genralrlization.

Description

A kind of antifatigue soluble granule and preparation method of the polypeptide dry powder containing Goat Placenta
Technical field
The present invention relates to a kind of Goat Placenta polypeptide soluble particles, digest Goat Placenta for molecule matter more particularly to a kind of Measure 1000 micromolecule active polypeptides or free amino acid below mixture Goat Placenta polypeptide powder preparation method and its can The preparation method of soluble particles.
Background technique
It has good health and a long life from ancient times to the present, continues life, perpetual youth always is the pursuit that the mankind never abandon, with the mankind The material progress of society continues to develop, and scientific and technological level is continuously improved, and more excites people and enjoys life, and explores the desire of life miracle It hopes.Delay senescence, build up health therewith, the functional food of beautifying face and moistering lotion comes into being, and have considerable economy Value.
Human plactnta also known as be dried human placenta, be a kind of common strengthening by means of tonics medicine, since ancient times by the successive dynasties name doctor reuse.One Before more than thousand years, the Chen Cangqi of the Tang Dynasty is recorded Human plactnta as tonic into supplement to the Herbal first.Hereafter Compendium of Material Medica Have to placenta recorded in the medicine ancient books such as " this is through meeting source ", claim: " it is warm-natured it is nontoxic, enter the warp of lung liver kidney three, it is main add essence, help gas, Beneficial blood, help it is empty, control men and women's consumptive disease, pledge one's devotion it is in a trance etc., to fatigue, it is thin, languisher have a special effect, long term usage person is clear-headed and clear-sighted, beard and hair It is black, promote longevity, have the effect of taking good fortune by force ".In Japan, Human plactnta in various Chinese prescription medicines, be commonly used to control consumptive disease, syntexis, Chronic tuberculosis, neurasthenia, anaemia, bronchial asthma, infertility, physiology have some setbacks, prevent miscarriage, anti-aging etc..It is modern Medicine also demonstrates immunoglobulin (Ig) rich in Human plactnta, active peptides, growth factor, phosphatide, polysaccharide, amino Acid and minerals, microelement isoreactivity substance.
Human plactnta is medically still applied at present, but because the limitation in ethics is disputed on there is extensive, Europe has been forbidden to use people's histoorgan and has used as drug, and Human plactnta Product Definition was also not support in 2000 by China, Therefore following to have fallen in the development and utilization to animal placenta.Although there are many placenta amount in China pig, to involve Some problems in religion, are unfavorable for the popularization of product；And the trophic component of Goat Placenta and Human plactnta are almost the same, trophic structure Closest to Human plactnta, become the preferred type of animal placenta.In addition Goat Placenta will not infect hepatitis virus, will not generate with the mankind Cross-infection, other animal placentas (including Human plactnta) are likely to this virus, relatively safe using Goat Placenta.Sheep Placenta raw material sources are abundant also to provide precondition for the further investigation of Goat Placenta and exploitation.
There are many preparation methods of placental article, including oven drying method, desivac, multigelation method, enzymatic isolation method, acid or buck Solution, organic solvent extraction, supercritical extraction method etc., wherein the method for extracting Goat Placenta polypeptide mainly has multigelation Method, acid or alkali hydrolysis method, enzymatic isolation method.
The process flow of multigelation method are as follows: placenta, which pre-processes, → shredding → is homogenized → multigelation → centrifugation → micro-filtration → Ultrafiltration.Wherein multigelation is a step of most critical in entire preparation process, it is desirable that by placenta homogenate in -30 DEG C or so of ring 24-48h is freezed under border, and needs multigelation 3-4 times.The method production cycle is long, at high cost, product yield is lower, is not easy to It is prepared by the mass production of Goat Placenta polypeptide.
The cardinal principle of acid or alkali hydrolysis method is to adjust pH value by acid or alkali, makes protein under the conditions of peracid or alkali excessively Denaturation, degradation occurs, to generate polypeptide or free amino acid.The method is due to having used strong acid, highly basic that can cause certain in placenta The loss of a little active materials, as acid hydrolyzation can cause the loss of whole tryptophans, part serine and tyrosine；Alkaline process can make ammonia Racemization occurs for base acid, inactivates threonine, arginine etc. largely.And the method corrosivity is strong, poor operability and produces Raw a large amount of waste liquids, can cause certain environmental pollution.
Enzyme hydrolysis method is exactly the specificity using enzyme, the certain peptides being selectively applied in protein under given conditions Key is allowed to be degraded into micromolecule polypeptide that is easy to dissolve and absorbing.The method reaction condition is mild, it is with short production cycle, at low cost, Rate is high, and the active material that can be effectively retained in Goat Placenta, is a kind of new technique efficiently, environmentally friendly, healthy.
Currently, domestic placenta based article mainly has: placental peptide injection, Goat Placenta soft capsule, Goat Placenta capsule, sheep placenta Disk oral solution, Goat Placenta shin moisturizer, Goat Placenta moisturizing and nourishing frost etc..
China is an animal husbandry big country, has a large amount of Goat Placentas to generate every year, however most of Goat Placenta is not able to Adequately development and utilization, cause the great wasting of resources.The value of the bioactive substance extracted from a Goat Placenta It is far longer than the value of a sheep itself, comprehensive development and utilization such as is carried out to Goat Placenta, can not only improve the receipts of local peasants and herdsmen Enter, increases local finance tax revenue, China's commonwealth physical constitution also can be improved, this will generate huge economic benefit and society It can benefit.
Summary of the invention
The technical problem to be solved by the invention is to provide one kind to be easy to absorb, the good Goat Placenta polypeptide powder of flavor taste The preparation method of sol particle.
To solve the above-mentioned problems, the technical scheme adopted by the invention is that:
A kind of preparation method of the soluble granule of Goat Placenta polypeptide dry powder, which is characterized in that specific step is as follows:
1) Goat Placenta raw material is pre-processed, carries out tissue homogenate；
2) two kinds and the above protease, enzymatic hydrolysis, the de- bitter and enzyme deactivation of deodorant are added in homogenate；
3) centrifugation removal of impurities, obtains Goat Placenta polypeptide solution；
4) it is concentrated and dried, obtains Goat Placenta polypeptide dry powder；
5) Goat Placenta polypeptide dry powder and synanthrin or xylose are mixed in proportion, granule is made.
Preferably, the protease of addition includes in the step 2)Protease, neutral proteinase and wind Taste protease.
Preferably, the constituent content of the protease are as follows:Protease is 60-80%, flavor protease For 20-30%, neutral proteinase 0-10%, the sum of each protease content is 100%.
Preferably, Goat Placenta pretreatment of raw material includes: to choose fresh Goat Placenta for raw material, described in addition in the step 1) 0.5~1.0 times of pure water tissue homogenate of fresh Goat Placenta weight.
Preferably, the step 2) the following steps are included:
A. after homogenate step 1) obtained heats 10~15min at a temperature of 90 DEG C, 50~55 DEG C are adjusted the temperature to；
B. the amount of 8~30ml protease is added by the fresh Goat Placenta of every 1kg, is added in obtained homogenateProtease, neutral proteinase and flavor protease, and controlled at 50~55 DEG C, enzyme 3.5-4.5h；
C. the enzymolysis liquid temperature that step B is obtained is adjusted to 30~35 DEG C；
D. the amount of 1.0~1.5g active dry yeast powder is added by the fresh Goat Placenta of every 1kg, in the enzymolysis liquid that step C is obtained Interior addition active dry yeast powder cultivates 60~90min；
E. enzymolysis liquid step D obtained heats 10~15min at 85~90 DEG C, carries out enzyme deactivation.
Preferably, the concentrate drying in the step 4) is that Goat Placenta polypeptide solution is concentrated into original under vacuum conditions The 1/3-1/2 of volume, is then spray-dried, and obtains Goat Placenta polypeptide dry powder.
Preferably, the preparation process of the step 5) is as follows:
Goat Placenta polypeptide dry powder is mixed with purified water, filler, being made by spray-drying process can Goat Placenta polypeptide The soluble granule of dry powder.
Preferably, sharp powder, soluble starch, dextrin, DEXTROSE ANHYDROUS, lactose, mannitol, mountain supplemented by the filler One of pears alcohol is a variety of.
It is another object of the present invention to provide a kind of Goat Placenta polypeptide powder sol particles.
To achieve the goals above, The technical solution adopted by the invention is as follows:
A kind of Goat Placenta polypeptide powder sol particle, including Goat Placenta polypeptide freeze-dried powder and filler, the Goat Placenta are more The ratio of peptide freeze-dried powder and the filler is 1:2-3.
Preferably, the filler is synanthrin.
A kind of preparation method of Goat Placenta polypeptide powder of the invention and its preparation method of sol particle are crossed and add wood A kind of sol particle is made in Goat Placenta polypeptide by sugar, synanthrin etc., is reduced the water imbibition of Goat Placenta polypeptide powder, is improved sheep placenta The water solubility of disk polypeptide powder and effectively improve product special flavour.
Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below There is attached drawing needed in technical description to be briefly described, it should be apparent that, the accompanying drawings in the following description is only this The embodiment of invention for those of ordinary skill in the art without creative efforts, can also basis The attached drawing of offer obtains other attached drawings.
Fig. 1 is the influence diagram of protease species and its enzyme concentration to peptide yield
Fig. 2 is the influence diagram of protease species and its enzyme concentration to peptide content
Influence diagram of Fig. 3 enzymolysis time to peptide yield and content
Influence diagram of Fig. 4 enzymolysis time to peptide relative molecular mass distribution
Influence diagram of Fig. 5 hydrolysis temperature to peptide yield and content
Influence diagram of Fig. 6 hydrolysis temperature to peptide relative molecular mass distribution
Influence diagram of Fig. 7 solid-liquid ratio to peptide yield and content
Influence diagram of Fig. 8 solid-liquid ratio to peptide relative molecular mass distribution.
Specific embodiment
The following is a clear and complete description of the technical scheme in the embodiments of the invention, it is clear that described embodiment Only a part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, the common skill in this field Art personnel every other embodiment obtained without making creative work belongs to the model that the present invention protects It encloses.
One, embodiment
Embodiment 1-4 is the present invention about a kind of antifatigue soluble granule of polypeptide dry powder containing Goat Placenta and preparation side The specific embodiment of method
Embodiment 1:
1) Goat Placenta pretreatment of raw material: choosing fresh Goat Placenta is raw material, is cleaned up, and removes impurity, and draining is weighed, 0.6 times of pure water of Goat Placenta weight is added and carries out tissue homogenate.
2) enzymatic hydrolysis and the de- hardship of deodorant: heating 10min at a temperature of 90 DEG C for above-mentioned homogenate, adjust the temperature to 50 DEG C, you The fresh Goat Placenta of every 1kg is added 6ml's afterwardsThe flavor protease of protease and 1ml, controlled at 50 DEG C, Digest 4h.Then enzymolysis liquid temperature is adjusted to 30 DEG C, the active dry yeast powder culture of 1.0g 60 minutes is added, finally by enzyme Liquid is solved in 90 DEG C of heating 10min, carries out enzyme deactivation.
3) centrifugation removal of impurities: above-mentioned enzymolysis liquid is subjected to centrifugation 10min using high speed low temperature centrifugal machine, removal water-insoluble obtains The weak yellow liquid of clear, i.e. Goat Placenta polypeptide solution.
4) it is concentrated and dried: Goat Placenta polypeptide solution being concentrated into the 1/2 of original volume under vacuum conditions, is then done by spraying It is dry, Goat Placenta polypeptide dry powder.
5) the quality control of Goat Placenta polypeptide:
1. it is more to calculate to determine nitrogen pool and free aminoacid content using kjeldahl apparatus and high performance liquid chromatograph The content of peptides of peptide content, this product must not be lower than 50%；
2. being measured using high performance gel filtration chromatography to Mr of peptide distribution, opposite point in this product Polypeptide of the protonatomic mass lower than 1000 must not be lower than the 80% of total hydrolysate.
6) preparation of soluble granule: Goat Placenta polypeptide dry powder and filler are mixed in the ratio of 1:2, particle is made Agent.
Embodiment 2:
1) Goat Placenta pretreatment of raw material: choosing fresh Goat Placenta is raw material, is cleaned up, and removes impurity, and draining is weighed, 0.6 times of pure water of Goat Placenta weight is added and carries out tissue homogenate.
2) enzymatic hydrolysis and the de- hardship of deodorant: heating 10min at a temperature of 90 DEG C for above-mentioned homogenate, adjust the temperature to 50 DEG C, you The fresh Goat Placenta of every 1kg is added 7ml's afterwardsThe flavor protease of protease and 2ml, controlled at 52 DEG C, digest 4h；
Then enzymolysis liquid temperature is adjusted to 30 DEG C, adds the active dry yeast powder culture of 1.0g 60 minutes, finally will Enzymolysis liquid carries out enzyme deactivation in 90 DEG C of heating 10min.
3) centrifugation removal of impurities: above-mentioned enzymolysis liquid is subjected to centrifugation 10min using high speed low temperature centrifugal machine, removal water-insoluble obtains The weak yellow liquid of clear, i.e. Goat Placenta polypeptide solution.
4) it is concentrated and dried: Goat Placenta polypeptide solution being concentrated into the 1/3 of original volume under vacuum conditions, is then done by spraying It is dry, Goat Placenta polypeptide dry powder.
5) the quality control of Goat Placenta polypeptide:
1. it is more to calculate to determine nitrogen pool and free aminoacid content using kjeldahl apparatus and high performance liquid chromatograph The content of peptides of peptide content, this product must not be lower than 50%；
2. being measured using high performance gel filtration chromatography to Mr of peptide distribution, opposite point in this product Polypeptide of the protonatomic mass lower than 1000 must not be lower than the 80% of total hydrolysate.
6) preparation of soluble granule: Goat Placenta polypeptide dry powder and filler are mixed in the ratio of 1:2.5, are made Granula.
Embodiment 3:
1) Goat Placenta pretreatment of raw material: choosing fresh Goat Placenta is raw material, is cleaned up, and removes impurity, and draining is weighed, 0.5 times of pure water of Goat Placenta weight is added and carries out tissue homogenate.
2) enzymatic hydrolysis and the de- hardship of deodorant: heating 10min at a temperature of 90 DEG C for above-mentioned homogenate, adjust the temperature to 54 DEG C, you The fresh Goat Placenta of every 1kg is added 7ml's afterwardsProtease, the flavor protease of 2ml and 1ml neutral proteinase, Controlled at 50 DEG C, 4h is digested；
Then enzymolysis liquid temperature is adjusted to 30 DEG C, adds the active dry yeast powder culture of 1.3g 60 minutes, finally will Enzymolysis liquid carries out enzyme deactivation in 90 DEG C of heating 10min.
3) centrifugation removal of impurities: above-mentioned enzymolysis liquid is subjected to centrifugation 10min using high speed low temperature centrifugal machine, removal water-insoluble obtains The weak yellow liquid of clear, i.e. Goat Placenta polypeptide solution.
4) it is concentrated and dried: Goat Placenta polypeptide solution being concentrated into the 1/2 of original volume under vacuum conditions, is then done by spraying It is dry, Goat Placenta polypeptide dry powder.
5) the quality control of Goat Placenta polypeptide:
1. it is more to calculate to determine nitrogen pool and free aminoacid content using kjeldahl apparatus and high performance liquid chromatograph The content of peptides of peptide content, this product must not be lower than 50%；
2. being measured using high performance gel filtration chromatography to Mr of peptide distribution, opposite point in this product Polypeptide of the protonatomic mass lower than 1000 must not be lower than the 80% of total hydrolysate.
6) preparation of soluble granule: Goat Placenta polypeptide dry powder and filler are mixed in the ratio of 1:2.8, are made Granula.
Embodiment 4:
1) Goat Placenta pretreatment of raw material: choosing fresh Goat Placenta is raw material, is cleaned up, and removes impurity, and draining is weighed, 1 times of pure water of Goat Placenta weight is added and carries out tissue homogenate.
2) enzymatic hydrolysis and the de- hardship of deodorant: heating 10min at a temperature of 90 DEG C for above-mentioned homogenate, adjust the temperature to 55 DEG C, you The fresh Goat Placenta of every 1kg is added 7ml's afterwardsProtease, the flavor protease of 2ml and 1ml neutral proteinase, Controlled at 50 DEG C, 4h is digested；
Then enzymolysis liquid temperature is adjusted to 30 DEG C, adds the active dry yeast powder culture of 1.3g 60 minutes, finally will Enzymolysis liquid carries out enzyme deactivation in 90 DEG C of heating 10min.
3) centrifugation removal of impurities: above-mentioned enzymolysis liquid is subjected to centrifugation 10min using high speed low temperature centrifugal machine, removal water-insoluble obtains The weak yellow liquid of clear, i.e. Goat Placenta polypeptide solution.
4) it is concentrated and dried: Goat Placenta polypeptide solution being concentrated into the 1/2 of original volume under vacuum conditions, is then done by spraying It is dry, Goat Placenta polypeptide dry powder.
5) the quality control of Goat Placenta polypeptide:
1. it is more to calculate to determine nitrogen pool and free aminoacid content using kjeldahl apparatus and high performance liquid chromatograph The content of peptides of peptide content, this product must not be lower than 50%；
2. being measured using high performance gel filtration chromatography to Mr of peptide distribution, opposite point in this product Polypeptide of the protonatomic mass lower than 1000 must not be lower than the 80% of total hydrolysate.
6) preparation of soluble granule: Goat Placenta polypeptide dry powder and filler are mixed in the ratio of 1:2.8, are made Granula.
In the above embodiment of the invention:
DescribedThe product that protease uses Royal DSM food ingredient (Shanghai) Co., Ltd. to go out；
The product that the flavor protease is gone out using letter Chinese biological Technology Co., Ltd. of Novi；
The product that the neutral proteinase is gone out using letter Chinese biological Technology Co., Ltd. of Novi；
The product that the active dry yeast powder uses Angel Yeast Co., Ltd to go out.
Two, for the measurement of process conditions
It is generally believed that influence hydrolysis result because being known as following seven kinds: pH, temperature, time, enzyme dosage, concentration of substrate, suppression Preparation, activator.Although inhibitor, activator can influence enzymolysis speed, uncertain factor can be generally introduced, is made at the later period Reason is difficult, and the considerations of for cost and foodsafety, the present invention is not explored the two factors.
The opening of peptide bond will lead to the decline of hydrolyzate pH value during protein digestion, therefore need outside constantly in hydrolytic process Add lye to maintain the pH value of hydrolyzate, it is ensured that enzyme is in always within the scope of its optimal pH, to improve the yield of polypeptide.But Because a large amount of salt for adding lye to generate will have a direct impact on the ionic strength and mouthfeel of product, the difficulty of product later period desalting processing is increased Degree is unfavorable for reducing product cost.In addition the nutritive value of protein can decrease under alkaline condition.Therefore, in the present invention All enzymatic hydrolysis are carried out under uncomfortable pH value state.
In order to make technological parameter optimized parameter of the invention, inventor has carried out further measurement, specific measuring method It is following several:
1) measurement of moisture content: gravimetric detemination is used
2) measurement of total protein content: Kjeldahl's method
3) peptide content measures: trichloroacetic acid precipitation combination biuret method
4) the relative molecular mass distribution measurement of peptide: gel filtration chromatography
5) peptide yield measures: water-soluble peptide dry weight/water-soluble peptide dry weight+precipitating dry weight
Test selection of the A. about protease
Respectively select Delvolase, Neutrase, Protamex, Flavourzyme, Trypsin, COLLUPLIN, Goat Placenta is hydrolyzed in 7 kinds of protease of validase, and the enzymatic hydrolysis condition of each enzyme is as shown in table 1.
Table 1
It tests in A with 0,1/10000,5/10000,1/1000,5/1000,1/100 (mL/g) enzyme concentration respectively to 7 kinds of enzymes It is determined, different protease gained enzymolysis product under the enzymatic hydrolysis condition of different enzyme concentrations is as shown in Figure 1.Fig. 1 is albumen The influence diagram of enzyme class and its enzyme concentration to peptide yield.
From figure 1 it appears that the enzyme concentration of different protease is almost the same on peptide yield influence, with the increasing of enzyme concentration Greatly, the yield of peptide also gradually increases.Wherein peptide yield highest in Delvolase protease hydrolyzed product, when enzyme concentration is 5/ When 1000 (mL/g), peptide yield is up to 71.88%, and validase protease is weaker to the hydrolysis of Goat Placenta, enzymolysis product Middle peptide yield is minimum, substantially 30% or so.
As shown in Fig. 2, Fig. 2 is the influence diagram of protease species and its enzyme concentration to peptide content.It can be seen from the figure that not Enzyme concentration with protease is larger on peptide content influence difference.Wherein in Delvolase protease hydrolyzed product peptide content compared with it His enzymolysis product is high, and under the hydrolysising condition of different enzyme concentrations, peptide content substantially remains in 55% or so；Flavourzyme egg Peptide content reaches highest (37.4%) when enzyme concentration is 5/10000 in white enzyme enzymolysis product, later with the increase of enzyme concentration, Peptide content gradually decreases instead；Peptide content is minimum in validase protease hydrolyzed product, substantially remains in 14% or so, with The increase of enzyme concentration, the molecular mass of peptide has the tendency that gradually decreasing in enzymolysis product, but after reaching a certain level, average molecular Mass Distribution is held essentially constant.
Test B: influence of the enzymolysis time to hydrolysis result
Goat Placenta is hydrolyzed using Delvolase protease, hydrolysising condition are as follows: solid-liquid ratio 1:1, enzyme dosage 5/ 1000, hydrolysis temperature is 50 DEG C, and enzymolysis time is followed successively by 1~8h.Distinguished using biuret reagent method and gel filtration chromatography Peptide content and relative molecular weight distribution are measured, studies influence of the enzymolysis time to Goat Placenta hydrolysis result accordingly.In enzymolysis product Peptide yield and assay result are as indicated at 3；Peptide relative molecular mass distribution is as shown in Figure 4.
From the figure 3, it may be seen that peptide yield has the tendency that gradually increasing with the extension of enzymolysis time.In a certain range with The extension of enzymolysis time, peptide content are gradually increased, content highest when 5~6h, but are extended as time go on, and peptide content starts Decline.It may be that the extension of enzymolysis time enhances Degree of Enzymatic Hydrolysis, produce a large amount of amino acid, and lower peptide content.
As shown in Figure 4, with the extension of enzymolysis time, the relative molecular mass of peptide has what is gradually decreased to become in enzymolysis product Gesture, but change more slow.
Test C: influence of the hydrolysis temperature to hydrolysis result
Goat Placenta is hydrolyzed using Delvolase protease, hydrolysising condition are as follows: solid-liquid ratio 1:1, enzyme dosage 5/ 1000, enzymolysis time 4h, hydrolysis temperature are respectively 40 DEG C, 45 DEG C, 50 DEG C, 55 DEG C, 60 DEG C.Using biuret reagent method and gel Filtration Chromatography measures peptide content and molecular weight distribution respectively, studies influence of the enzymolysis time to Goat Placenta hydrolysis result accordingly, As figure 5 illustrates, peptide relative molecular mass distribution is as shown in Figure 6 for peptide yield and content experimental result in enzymolysis product.
As shown in Figure 5, peptide yield is gradually increased as the temperature rises, but peptide content be in substantially 50%~55% it Between, variation radian is smaller.It will be appreciated from fig. 6 that as the temperature rises, peptide relative molecular mass has gradually downward trend, still Change more slow.
Test D: influence of the solid-liquid ratio to hydrolysis result
Goat Placenta is hydrolyzed using Delvolase protease, hydrolysising condition are as follows: enzyme dosage 5/1000, when enzymatic hydrolysis Between 4h, 50 DEG C of hydrolysis temperature, solid-liquid ratio is respectively 2:1,1:1,1:2,1:3,1:4.Using biuret reagent method and gel filtration Chromatography measures peptide content and molecular weight distribution respectively, studies influence of the solid-liquid ratio to Goat Placenta hydrolysis result accordingly, and enzymatic hydrolysis produces As shown with 7, peptide relative molecular mass distribution is as shown in Figure 8 for peptide yield and content experimental result in object.
As shown in Figure 7, with the reduction that the reduction of solid-liquid ratio is concentration of substrate, peptide yield can be increased to a certain extent Height, but it is substantially unchanged lower than after 1:2.Less, many places are between 50%~55% for peptide content variation, it can thus be seen that material Liquor ratio smaller is influenced on hydrolysis result from figure 8, it is seen that with solid-liquid ratio reduction, peptide relative molecular mass keeps substantially It is constant, illustrate in a certain range, influence of the solid-liquid ratio to Delvolase protease hydrolyzed Goat Placenta is smaller.
In conclusion albumen enzyme in the factors such as the type and its enzyme concentration of enzyme, enzymolysis time, hydrolysis temperature, solid-liquid ratio Class has a significant impact hydrolysis result, it follows that technical solution of the present invention, which passes through, chooses suitable protease, and openly The optimum hydrolysising condition of reasonably optimizing makes entire preparation process be able to achieve preferably preparation effect.
Three, anti-fatigue performance is tested
1, the design of animal packet and given low
When using serum urea and hepatic glycogen content as index, take balanced randomized blocks small by 60 according to mouse weight Mouse is divided into 3 groups, i.e. Goat Placenta-peptide group, soybean peptide group, blank control group, and every group 20.
When using the mice burden swimming time as testing index, choose first time walking weight load be in 40~80min it Between 30 mouse, then take balanced randomized blocks that mouse is divided into Goat Placenta-peptide group, soybean according to walking weight load Peptide group, blank control group, every group 10.
Intragastric administration on mice dosage is Goat Placenta-peptide group (500mg/kg/d), soybean peptide group (500mg/kg/d), blank control group (isometric physiological saline).
Adaptive feeding starts stomach-filling after 7 days, the laundering period gives normal diet, gives Low protein diet after administration (40% corn flour, 40% wheat-middlings, 20% wheat bran, separately add a small amount of vitamin A, vitamin B1, vitamin C and edible salt). Continuously give tested material 15 days.
2, the measurement of mouse weight
Mouse records a weight every three days before administration, after administration, calculates the average value that each group mouse is weighed every time, makees Record.
3, swimming with a load attached to the body is tested
Before administration, two weeks same day after latter week is administered, is administered, after last gives given the test agent 30min, mouse tail Mouse is put into depth of water 30cm, swum in the swimming pool that 25 ± 1 DEG C of water temperature by the fuse of its 7% weight of weight of load.Note The time that cannot be emerged mouse is recorded since in investment water to head whole submerged 10s, as mice burden swimming Time.
4, result
Influence of the Goat Placenta-peptide to mouse weight is as shown in following table table 2:
2 Goat Placenta-peptide of table to mouse weight influence (N=10)
Control group and the tested 15 days weight growth patterns of experimental mice can be seen that control group and reality from upper table table 2 The mouse weight for testing group generally all shows a increasing trend, and growth pattern slightly has difference.To the weight of each group mouse after administration 15 days Carry out one-way analysis of variance, F value=0.010, F0.05(2,27)=3.35, F value < F0.05, illustrate blank control group, soybean peptide There was no significant difference between group, Goat Placenta-peptide group.The above result shows that Goat Placenta-peptide does not make significant difference to mouse weight.
Influence of the Goat Placenta-peptide to the mice burden swimming time is as shown in following table table 3:
3 Goat Placenta-peptide of table to the mice burden swimming time influence (N=10)
Note: * * expression is compared with blank control group, P < 0.01, and difference is extremely significant.
Blank control group and experimental mice administration front and back walking weight load can be seen that blank pair from upper table table 3 It generally all shows a increasing trend according to the mice burden swimming time of group and experimental group, but experimental group is compared with blank control group increasing degree Greatly.To administration one week and after two weeks between each group the mice burden swimming time carries out one-way analysis of variance respectively, after a week F value= 5.835, F value > F0.05, have extremely significant sex differernce (P < 0.01) between blank control group and Goat Placenta-peptide group, after two weeks F value= 4.108, F value > F0.05, there is significant difference (P < 0.01) between blank control group and Goat Placenta-peptide group.The above result shows that Goat Placenta-peptide has conspicuousness accretion to the mice burden swimming time, and thus deducing Goat Placenta-peptide has centainly antifatigue Activity.
Influence of the Goat Placenta-peptide to serum urea and hepatic glycogen content is as shown in following table table 4:
4 Goat Placenta-peptide of table to serum urea and hepatic glycogen content influence (N=10)
Group Serum urea content (mmoL/L) Hepatic glycogen content (mg/g)
Blank control group 9.08±0.54 10.45±1.76
Soybean peptide group 7.86±0.69* 11.77±1.77
Goat Placenta-peptide group 6.90±1.10** 11.89±2.90
Note: * expression is compared with blank control group, P < 0.05, significant difference；* indicate compared with blank control group, P < 0.01, difference is extremely significant.
By upper table table 4 it is found that experimental mice post exercise serum urea content is significantly lower than blank control group, to each group Between mice serum urea content carry out one-way analysis of variance, F value=10.837, F value > F0.05, blank control group and soybean peptide There is significant difference (P < 0.05) between peptide group, there is extremely significant sex differernce (P < between blank control group and Goat Placenta-peptide group 0.01), illustrate the function of serum urea content after Goat Placenta-peptide and soybean peptide are reduced mouse movement.
The length of mice burden swimming time can intuitively reflect the degree of fatigue of mouse, and swimming experiment device letter The single, physical efficiency convenient and that body can be objectively responded of operation, therefore become and measure most common evaluation index in anti-fatigue active.Sheep Placenta peptide can be obviously prolonged the walking weight load of mouse, reduce the serum urea content of mouse after movement, and with blank pair According to group compared to tool significant difference, Goat Placenta-peptide is thus deduced with certain anti-fatigue active.
Each embodiment in this specification is described in a progressive manner, the highlights of each of the examples are with other The difference of embodiment, the same or similar parts in each embodiment may refer to each other.For device disclosed in embodiment For, since it is corresponded to the methods disclosed in the examples, so being described relatively simple, related place is said referring to method part It is bright.
The foregoing description of the disclosed embodiments enables those skilled in the art to implement or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, as defined herein General Principle can be realized in other embodiments without departing from the spirit or scope of the present invention.Therefore, of the invention It is not intended to be limited to the embodiments shown herein, and is to fit to and the principles and novel features disclosed herein phase one The widest scope of cause.

Claims (10)

1. a kind of preparation method of the antifatigue soluble granule of polypeptide dry powder containing Goat Placenta, which is characterized in that specific steps It is as follows:
1) Goat Placenta raw material is pre-processed, carries out tissue homogenate；
2) two kinds and the above protease, enzymatic hydrolysis, the de- bitter and enzyme deactivation of deodorant are added in homogenate；
3) centrifugation removal of impurities, obtains Goat Placenta polypeptide solution；
4) it is concentrated and dried, obtains Goat Placenta polypeptide dry powder；
5) Goat Placenta polypeptide dry powder and synanthrin or xylose are mixed in proportion, granule is made.
2. a kind of preparation method of the antifatigue soluble granule of polypeptide dry powder containing Goat Placenta according to claim 1, It is characterized in that, the protease being added in the step 2) includesProtease, neutral proteinase and flavor egg White enzyme.
3. a kind of preparation method of the antifatigue soluble granule of polypeptide dry powder containing Goat Placenta according to claim 2, It is characterized in that, the constituent content of the protease are as follows:Protease is 60-80%, flavor protease 20- 30%, neutral proteinase 0-10%, the sum of each protease content are 100%.
4. a kind of preparation method of the antifatigue soluble granule of polypeptide dry powder containing Goat Placenta according to claim 3, It is characterized in that, Goat Placenta pretreatment of raw material includes: to choose fresh Goat Placenta as raw material in the step 1), it is added described fresh 0.5~1.0 times of pure water tissue homogenate of Goat Placenta weight.
5. a kind of preparation method of the antifatigue soluble granule of polypeptide dry powder containing Goat Placenta according to claim 3, It is characterized in that, the step 2) the following steps are included:
A. after homogenate step 1) obtained heats 10~15min at a temperature of 90 DEG C, 50~55 DEG C are adjusted the temperature to；
B. the amount of 8~30ml protease is added by the fresh Goat Placenta of every 1kg, is added in obtained homogenate Protease, neutral proteinase and flavor protease, and controlled at 50~55 DEG C, enzyme 3.5-4.5h；
C. the enzymolysis liquid temperature that step B is obtained is adjusted to 30~35 DEG C；
D. the amount of 1.0~1.5g active dry yeast powder is added by the fresh Goat Placenta of every 1kg, adds in the enzymolysis liquid that step C is obtained Enter active dry yeast powder, cultivates 60~90min；
E. enzymolysis liquid step D obtained heats 10~15min at 85~90 DEG C, carries out enzyme deactivation.
6. a kind of preparation method of the antifatigue soluble granule of polypeptide dry powder containing Goat Placenta according to claim 1, It is characterized in that, the concentrate drying in the step 4) is, Goat Placenta polypeptide solution is concentrated into original volume under vacuum conditions 1/3-1/2, be then spray-dried, obtain Goat Placenta polypeptide dry powder.
7. a kind of preparation method of the antifatigue soluble granule of polypeptide dry powder containing Goat Placenta according to claim 1, It is characterized in that, the preparation process of the step 5) is as follows:
Goat Placenta polypeptide dry powder is mixed with purified water, filler, being made by spray-drying process can Goat Placenta polypeptide dry powder Soluble granule.
8. a kind of preparation method of the antifatigue soluble granule of polypeptide dry powder containing Goat Placenta according to claim 7, It is characterized in that, sharp powder, soluble starch, dextrin, DEXTROSE ANHYDROUS, lactose, mannitol, sorbierite supplemented by the filler One of or it is a variety of.
9. requiring a kind of antifatigue sol particle of polypeptide dry powder containing Goat Placenta of any preparation method preparation of 1-8 according to power Agent, which is characterized in that including Goat Placenta polypeptide freeze-dried powder and filler, the Goat Placenta polypeptide freeze-dried powder and the filler Ratio is 1:2-3.
10. a kind of antifatigue soluble granule of the polypeptide dry powder containing Goat Placenta according to weighing and require 9, which is characterized in that The filler is synanthrin.
CN201810785568.7A 2018-07-17 2018-07-17 A kind of antifatigue soluble granule and preparation method of the polypeptide dry powder containing Goat Placenta Pending CN109222110A (en)

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CN1537465A (en) * 2003-10-14 2004-10-20 大连轻工学院 Functional food contg. ewe placenta, and its prepn. method
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CN113855731A (en) * 2021-09-27 2021-12-31 浙江康德药业集团股份有限公司 Oral liquid with antifatigue and immunoregulation functions and preparation method thereof
CN113855731B (en) * 2021-09-27 2023-01-17 浙江康德药业集团股份有限公司 Oral liquid with antifatigue and immunoregulation functions and preparation method thereof

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