CN112048417A - Active small-molecule donkey-hide gelatin paste and preparation method thereof - Google Patents
Active small-molecule donkey-hide gelatin paste and preparation method thereof Download PDFInfo
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- CN112048417A CN112048417A CN202010733397.0A CN202010733397A CN112048417A CN 112048417 A CN112048417 A CN 112048417A CN 202010733397 A CN202010733397 A CN 202010733397A CN 112048417 A CN112048417 A CN 112048417A
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Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
- C12G3/00—Preparation of other alcoholic beverages
- C12G3/04—Preparation of other alcoholic beverages by mixing, e.g. for preparation of liqueurs
- C12G3/05—Preparation of other alcoholic beverages by mixing, e.g. for preparation of liqueurs with health-improving ingredients, e.g. flavonoids, flavones, polyphenols or polysaccharides
-
- 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
- A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
- A23L13/20—Meat products; Meat meal; Preparation or treatment thereof from offal, e.g. rinds, skins, marrow, tripes, feet, ears or snouts
-
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Abstract
The invention discloses an active small-molecule donkey-hide gelatin paste and a preparation method thereof. The invention adopts donkey hide as an initial raw material, and prepares the active small molecular donkey-hide gelatin paste in the form of viscous liquid paste with the solid content of more than 40Brix for the first time. The donkey hide after treatment is made into donkey-hide gelatin paste through processes of soaking, extracting, filtering, separating liquid, enzymolysis, enzyme deactivation, concentration and the like, thereby innovating the 'drying and blocking' process which has the greatest difficulty, the highest cost and the longest time consumption in the traditional donkey-hide gelatin block preparation process, and greatly reducing the production cost. Meanwhile, the prepared active small molecular donkey-hide gelatin paste is viscous liquid paste, can be taken instantly at any time and any place, and solves the problem that the traditional donkey-hide gelatin blocks are complicated to take. Moreover, the content of the donkey-hide gelatin in the product is far higher than dosage forms such as donkey-hide gelatin cake and donkey-hide gelatin slurry, and the quality and the efficacy of the product are ensured. In addition, the molecular weight of the donkey-hide gelatin peptide is mainly distributed between 180-2000, so that the donkey-hide gelatin peptide is easier to absorb by a human body.
Description
Technical Field
The invention discloses active small-molecule donkey-hide gelatin paste and a preparation method thereof, belonging to the field of deep processing of donkey-hide gelatin.
Background
The traditional donkey-hide gelatin is a gelatin block prepared by decocting and concentrating the skin of donkey of an equine animal, adding auxiliary materials such as soybean oil, rock sugar and the like, has the functions of enriching blood and stopping bleeding, and is a famous traditional Chinese medicine. The main components of the food are collagen, peptide and amino acid except for containing a small amount of trace elements, wherein the protein component accounts for about 80 percent. In the process of preparing the donkey-hide gelatin, the process of drying and blocking is the longest in time consumption, the greatest in difficulty and the highest in cost, which is also the important reason for the high price and difficult popularization of the donkey-hide gelatin. The cost expenditure of this section is of no value in terms of efficacy. The invention creatively innovates a non-valuable block making process by taking the traditional paste as a carrier, and reduces the production cost.
Meanwhile, the traditional donkey-hide gelatin blocks are hard, poor in taste and tedious to eat, and are important factors limiting the circulation of the donkey-hide gelatin blocks. Although some preparations such as donkey-hide gelatin syrup, donkey-hide gelatin oral liquid and donkey-hide gelatin cake are available, the dosage form is limited by the carrier, and the donkey-hide gelatin content is generally low, so that the efficacy is limited. The method disclosed by the invention has the advantages that the content of the donkey-hide gelatin solid is more than 40Brix, the donkey-hide gelatin solid is a viscous liquid paste, the donkey-hide gelatin solid can be instantly taken at any time and any place, and the problem that the traditional donkey-hide gelatin blocks are complicated to take is solved.
In addition, the macromolecular protein component in the donkey-hide gelatin accounts for 80 percent, so the donkey-hide gelatin is difficult to digest and absorb, namely 'nourishing and greasy and obstructing the stomach' indicated by the traditional Chinese medicine. Therefore, people with poor digestion function have difficulty in exerting due efficacy when eating donkey-hide gelatin, and the gastrointestinal burden is increased. This is because large protein molecules cannot be directly absorbed by the human body, and after entering the digestive system, they are first hydrolyzed into polypeptides by pepsin in the stomach, and then further hydrolyzed into small peptides or amino acids by trypsin, aminopeptidase, chymotrypsin, etc. in the intestinal tract, and then absorbed by the human body.
In recent years, many small-molecule peptides (3-15 peptides, molecular weight below 2000) have been found to have specific pharmacological activity. For example, pentapeptide (Tyr-Ile-Gly-Ser-Arg) has activity against cancer cell metastasis; the dodecapeptide (Ile-Tyr-Leu-Gly-Gly-Pro-Phe-Ser-Pro-Asn-Val-Leu-NH2) has immunity regulating and anticancer effects. The collagen is hydrolyzed into high-activity small molecular peptide in vitro, so that the utilization rate of the human body can be greatly improved, the activity is increased, and the physical burden is reduced.
Therefore, the preparation of the liquid paste form instant food and high-absorption active small molecular donkey-hide gelatin paste with low cost has important significance.
In recent years, some reports on the preparation of small-molecular donkey-hide gelatin peptides exist, but expensive donkey-hide gelatin blocks or donkey-hide gelatin juice are used as raw materials, different types of protease are used for catalytic reaction, and the molecular weight distribution is 200-10000 Da. Because of the problems of solubility, etc., the dosage forms are mainly peptide granules and peptide powder. For example:
the invention discloses a method for preparing liquid donkey-hide gelatin by an enzyme hydrolysis method (publication number is CN 1237421A). in the method, donkey-hide gelatin blocks are used as raw materials, after being dissolved by adding water, the donkey-hide gelatin blocks are hydrolyzed by papain, bromelain or ficin to prepare liquid donkey-hide gelatin oral liquid, but the molecular weight range is not mentioned, and the solid content is not increased.
The invention patent of mixture of active micromolecule donkey-hide gelatin and preparation method and application thereof (publication number is CN102499376B) discloses a method, which takes donkey-hide gelatin juice as raw material, adopts compound protease including proline protease to carry out enzymolysis on the donkey-hide gelatin juice to prepare zymolyte rich in small peptide, and the peptide sections are distributed between 200 and 3000Da and mainly between 200 and 1000. The method does not consider the control of flavor, does not control residual oil, and does not solve the problems of high cost and convenient taking.
The invention discloses a micromolecule donkey-hide gelatin pure powder tablet and a preparation method thereof (publication number is CN104107192A), which take expensive donkey-hide gelatin blocks as raw materials, add water, heat and melt the gelatin, then hydrolyze the gelatin by one or more of papain, bromelain, neutral protease, alkaline protease and collagen hydrolase, and finally obtain a finished product after drying and tabletting. The method does not relate to the control of exposed amino acid and residual grease, and does not solve the problems of high cost and convenient eating.
The invention discloses a preparation method of low-bitter donkey-hide gelatin oligopeptide (publication number CN107028176A), which is used for preparing the low-bitter donkey-hide gelatin oligopeptide by embedding the donkey-hide gelatin oligopeptide with sodium alginate and arabic gum microcapsules, but the taste of the donkey-hide gelatin oligopeptide is not changed substantially, and the problems of high cost and convenience in eating are not solved.
The invention discloses a preparation method of donkey-hide gelatin oligopeptide (publication number is CN103122363B), which takes donkey-hide gelatin stock solution as raw material, heats and stirs, adjusts pH, adds Alcalase2.4L FG hydrolytic proteinase for hydrolysis, and finally sprays and dries. The method adopts single enzyme for hydrolysis, the addition amount of the enzyme is large, the pH value needs to be adjusted in the reaction process, the reaction time is long, oil is removed by a filtration method, and the control of flavor and the control of cost are not considered.
The methods use colla corii asini blocks or colla corii asini juice as raw materials to prepare the small-molecular colla corii asini peptide, and the control of the proteolytic sites is not considered, so that when the common protease hydrolyzes the protein, sulfur-containing amino acid (cystine and methionine) and basic amino acid (lysine) are exposed, odor and ammonia smell are generated, and the small-molecular colla corii asini peptide is difficult to swallow and causes discomfort. If the amount of sulfur-containing amino acids and basic amino acids exposed during hydrolysis is avoided or reduced, the odor and ammonia odor can be significantly reduced, and the taste can be improved. At present, no report of preparing small molecular donkey-hide gelatin peptide by a method for controlling a protein hydrolysis site is found; moreover, the existing small-molecule donkey-hide gelatin peptides are all prepared into powder or tablet products, and no report of preparing small-molecule donkey-hide gelatin paste is found.
Disclosure of Invention
Aiming at the existing problems, the invention provides a preparation method of a micromolecule donkey-hide gelatin paste with low cost, good flavor, convenient taking and high activity by taking donkey hide as an initial raw material and taking a traditional paste formula as a formulation carrier. In order to achieve the purpose, the invention adopts the following technical means:
the active small molecule donkey-hide gelatin paste is in a viscous liquid paste form, and the solid content is more than 40 Brix. The weight average molecular weight is 600-1500, and the peptide segment is mainly distributed between 180-2000 Da.
The preparation method of the active small-molecule donkey-hide gelatin paste is characterized by comprising the following steps:
s1 pretreatment: processing donkey skin into strip, block or granule, soaking, and cleaning.
S2 extraction: adding 2-20 times of clear water by weight into the processed donkey skin, and stirring and extracting or refluxing for extraction. The extraction temperature is 70-150 ℃, the stirring speed is 10-60 rpm, and the reflux speed is 1-6T/h. The extraction time is 11-73 h.
S3 filtering: extracting until the collagen is completely dissolved, filtering to remove insoluble impurities, and collecting the filtrate.
S4 liquid separation: standing the filtrate in an aseptic environment after sterilizing, wherein the standing temperature is 8-68 ℃, and the standing time is 3-24 hours; after layering, the upper oil layer was removed and the lower clear liquid was collected.
S5 enzymolysis: adding compound protease into the clear liquid for enzymolysis reaction. The enzymolysis time is 0.4-6.5h, and the reaction temperature is 40-75 ℃. After the reaction is finished, heating to over 90 ℃ to inactivate the enzyme.
S6 concentrating and making into paste: concentrating under reduced pressure at 55-75 deg.C and vacuum degree of-0.001-0.2 Mp until the solid content is above 40Brix, and making into active small molecule colla Corii Asini paste. After filling, the finished product can be prepared.
A preparation method of active small molecule donkey-hide gelatin paste is characterized in that the compound protease comprises metalloprotease and serine protease. One or more of subtilisin, acid protease, alkaline protease, neutral protease, glycosidase, papain, and trypsin can also be contained. The added metal protease and serine protease are 0.1-4% of the dry weight of the raw materials; and the added subtilisin, acid protease, alkaline protease, neutral protease, glycosidase, papain and trypsin are 0-2% of the dry weight of the raw materials.
A preparation method of active small molecule donkey-hide gelatin paste is characterized in that in step S1, an alkali solution with the weight of 0.01-3% of that of clear water can be added to accelerate foaming.
A preparation method of active small molecule donkey-hide gelatin paste is characterized in that the molecular weight of peptide segments is distributed between 180-3000Da and is more than 88 percent, and the molecular weight of peptide segments is distributed between 180-2000Da and is more than 82 percent.
A method for preparing small molecule colla Corii Asini paste comprises adding adjuvants in the concentration step. The auxiliary material is one or more of wine, alcohol, sugar and oligosaccharide prebiotics.
Drawings
FIG. 1 HPLC chromatogram of small molecule donkey-hide gelatin
FIG. 2 HPLC chromatogram of small molecule donkey-hide gelatin
FIG. 3 HPLC chromatogram of small molecule donkey-hide gelatin
FIG. 4 HPLC chromatogram of small molecule donkey-hide gelatin
Detailed Description
The technical solution of the present invention is further illustrated by some specific embodiments below:
sources of experimental materials: donkey hide, purchased by farmers.
Enzyme: metalloprotease, serine protease, subtilisin, acid protease, alkaline protease, neutral protease, glycosidase, papain, Novoxil (China) Biotechnology Inc.
Donkey-hide gelatin, baodingxiao donkey-hide gelatin limited.
And (3) detecting the molecular weight of the polypeptide by an HPLC method:
the instrument comprises the following steps: waters high performance liquid chromatograph e2695 equipped with diode array detector (PDA) 2998.
A chromatographic column: TSK gel G2000 SWXL 7.8mm X300 mm gel column.
Mobile phase: acetonitrile: water: trifluoroacetic acid 45:55:0.1
Detection wavelength: UV220nm
Flow rate: 0.5mL/min
Column temperature: 30 deg.C
Sample introduction volume is 10 mu L
Relative molecular weight calibration curve standard: cytochrome C (12500Da), phthalidase (6500Da), bacitracin (1450Da), ethanamine-tyrosine-arginine (451Da), ethanamine-ethanamine (189Da)
Example 1.
Taking 1kg donkey skin, washing with water, soaking for 2h, and soaking for hair. After cleaning and draining, 20kg of clean water is added, heating and stirring extraction are carried out at 90 ℃, the stirring speed is 25rpm, and the water loss is supplemented in the boiling process. After extraction for 58h, insoluble impurities were removed by filtration and placed in a separatory funnel. Standing at room temperature for 3h, and separating and removing the upper oily impurity layer. Heating the lower layer solution to 60 ℃, adding 2g of serine protease and 2g of metal protease, and carrying out enzymolysis by keeping the temperature and stirring for 3 hours. Heating to 98 deg.C, maintaining for 15min to inactivate enzyme, and concentrating under reduced pressure the small molecule colla Corii Asini hydrolysate to 60Brix to obtain active small molecule colla Corii Asini paste. The molecular weight is measured by HPLC, as shown in FIG. 1, the molecular weight of the peptide segment is 90.05% between 180 and 3000Da, and the weight average molecular weight is about 728.3 Da.
FIG. 1 HPLC detection spectrum of small molecule donkey-hide gelatin paste
Example 2.
Taking 100kg donkey skin, washing with water, cutting into pieces, and putting into an extraction tank. 100g of sodium hydroxide is weighed, dissolved in 300L of water and added into an extraction tank for foaming. After soaking, washing with clear water until the pH of the water washing solution is below 7. 2 tons of water are added, and the reflux extraction is carried out at 105 ℃, the reflux speed is 1T/h, and the extraction time is 15 hours. Filtering to remove insoluble impurities, transferring the filtrate to a liquid separation tank, sterilizing, adjusting the temperature to 40 ℃, standing for 4 hours, separating and removing an upper oily impurity layer, and collecting a lower clear liquid. Regulating the temperature to 56 ℃, adding 200g of serine protease, 100g of metalloprotease, 20g of neutral protease and 20g of papain in the mass of the donkey skin, stirring and reacting at constant temperature for 3 hours. Heating to 98 deg.C, maintaining for 15min to inactivate enzyme; then, concentrating the micromolecule donkey-hide gelatin hydrolysate after enzyme deactivation to 40Brix under the conditions that the temperature is 55 ℃ and the vacuum degree is-0.2 Mp. Adding 1kg of yellow wine and 1kg of rock sugar, stirring uniformly, and heating at 100 ℃ for 1h to obtain the active micromolecule donkey-hide gelatin paste. The molecular weight is detected by HPLC method, the molecular weight of the peptide segment is 92.2 percent distributed between 180 and 3000Da, and the weight average molecular weight is about 575.2 Da.
FIG. 2 HPLC detection spectrum of small molecule donkey-hide gelatin paste
Example 3.
Taking 100kg donkey skin, washing with water, cutting into pieces, and putting into an extraction tank. After foaming, 400kg of water was added, and extraction was carried out at 70 ℃ with stirring at 30rpm for 73 hours. Filtering to remove insoluble impurities, transferring the filtrate to a liquid separation tank, sterilizing, adjusting the temperature to 10 ℃, standing for 20 hours, separating and removing an upper oily impurity layer, and collecting a lower clear liquid. Adjusting the temperature to 60 ℃, adding 3kg of metalloprotease, 2kg of serine protease, 200g of subtilisin, 200g of acid protease, 300g of neutral protease and 300g of glycosidase, stirring and reacting at constant temperature for 3 h. Heating to 95 deg.C, maintaining for 15min to inactivate enzyme; then, the micromolecule donkey-hide gelatin hydrolysate after enzyme deactivation is decompressed and concentrated to 60Brix under the conditions that the temperature is 73 ℃ and the vacuum degree is-0.01 Mp.
Then 4kg of yellow wine, 50kg of fructo-oligosaccharide and 50kg of isomaltooligosaccharide are added and stirred evenly to obtain the active micromolecule donkey-hide gelatin paste. The molecular weight is detected by HPLC method, the molecular weight of the peptide segment is distributed between 180 and 3000Da and accounts for 90.01 percent, and the weight average molecular weight is about 740.7 Da.
FIG. 3 HPLC detection spectrum of small molecule donkey-hide gelatin paste
Example 4.
Taking 300kg of fresh donkey skin as a raw material, putting the raw material into an extraction tank, adding water, stirring, cleaning, and adding water again for soaking. Adding water again for 1T, heating to 90 deg.C, stirring and extracting at stirring speed of 58rpm for 12 hr. Removing insoluble impurities by filtering for multiple times with 30-200 meshes, and transferring to a liquid separation tank. Sterilizing, standing at 40 deg.C for 25 hr, separating oil layer from colla Corii Asini stock solution, removing oil layer, and collecting supernatant. Regulating the temperature to 60 ℃, adding 600g of metalloprotease and 600g of serine protease, and stirring for 3 hours at constant temperature. Then the temperature is raised to 105 ℃, and the enzyme is kept for 12min to be inactivated. Then, concentrating the micromolecule donkey-hide gelatin hydrolysate after enzyme deactivation under reduced pressure to 65Brix under the conditions that the temperature is 60 ℃ and the vacuum degree is-0.03 Mp. The active micromolecular donkey-hide gelatin paste is obtained.
The molecular weight is detected by HPLC method, the molecular weight of the peptide segment is distributed between 180 and 3000Da and accounts for 90.03 percent, and the weight average molecular weight is about 771.1 Da.
FIG. 4 HPLC detection spectrum of small molecule donkey-hide gelatin paste
Experimental example 1
The blood enriching effect of the small molecule donkey-hide gelatin paste is detected by taking the level of mouse hemoglobin (Hb) and red blood cell count (RBC) as investigation indexes.
Experimental materials: the small molecule donkey-hide gelatin paste prepared in example 4 herein; non-hydrolyzed colla Corii Asini block.
The experimental method comprises the following steps: preparing small molecular donkey-hide gelatin paste into 65Brix, 35Brix and 15Brix, and dissolving donkey-hide gelatin blocks in water to obtain 65Brix, 35Brix and 15Brix donkey-hide gelatin stock solutions. After 64 KM mice with the weight of 20-25 g and half of male and female are taken for adaptive feeding for 7d in an environment with the temperature of 20-22 ℃ and the relative humidity of 45-65%, the KM mice are randomly divided into 8 groups, wherein each group comprises 8 mice, 1 group is a blank group, and 7 groups are hematopoietic blood deficiency models. Mice in each group were bled at the tail before the experiment to determine normal hemoglobin (Hb) and red blood cell count (RBC). The tail of a mouse in the blood deficiency model group is bled by 0.5mL, Hb and RBC are measured 24h after bleeding, then 0.5mL of test solution with each concentration is administered, wherein 0.5mL of physiological saline is administered to the model building group, 0.5mL of physiological saline is administered to the blank group, 1 time of administration is performed every day, the mouse takes food 4h after administration, 15 days are continued, and blood is taken from the abdominal cavity vein in the next day after the last administration to measure Hb and RBC. The results are shown in table 1:
TABLE 1 Effect of Small molecule donkey-hide gelatin paste on blood deficiency mouse model caused by tail bleeding: (n=8)
Note: p is less than 0.05 compared with the molding group; compared with the same dosage group of colla Corii Asini stock solution, # P is less than 0.05
The experimental result shows that the micromolecular donkey-hide gelatin paste can obviously improve the Hb and RBC levels of a blood deficiency model mouse, and is obviously higher than the donkey-hide gelatin stock solution.
Experimental example 2
And detecting the influence of the small-molecule donkey-hide gelatin paste on the model mouse with cyclophosphamide-induced leukopenia by taking white blood cell count (WBC), RBC and Hb as research indexes. Preparing small molecular donkey-hide gelatin paste into 65Brix, 35Brix and 15Brix, and dissolving donkey-hide gelatin blocks in water to obtain 65Brix, 35Brix and 15Brix donkey-hide gelatin stock solutions. 64 KM mice with the weight of 20-25 g and half male and female are taken for 5-6 weeks, after the mice are bred adaptively for 7 days in an environment with the temperature of 20-22 ℃ and the relative humidity of 45-65%, the mice are randomly divided into 8 groups, wherein 1 group is a blank group, 0.5mL of normal saline is respectively filled in the blank group and a building module, 0.5mL of high, medium and low dose test solution is respectively filled in a donkey-hide gelatin stock solution group and a small molecular donkey-hide gelatin paste group, the administration is carried out for 1 time every day for 15 days continuously, from the 1 st day of the administration, except the blank group, each mouse is subjected to intraperitoneal injection of cyclophosphamide solution according to the amount of 40mg/kg, the injection is carried out for 3 days continuously, and after the administration is carried out for 2 hours for the last time, blood is taken from the tail of the mouse to determine white blood cell count (WBC), RBC and Hb, and.
TABLE 2 Effect of donkey-hide gelatin enzymolysis liquid on cyclophosphamide-induced leukopenia model in mice: (n=8)
Note: p is less than 0.05 compared with the molding group; compared with the same dosage group of colla Corii Asini stock solution, # P is less than 0.05
From the results in table 2, it can be seen that the small molecule donkey-hide gelatin paste has a significant effect of improving the WBC, RBC and Hb levels of mice with leukopenia caused by cyclophosphamide, and the effect is significantly higher than that of donkey-hide gelatin stock solution.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the scope of the present invention, and all modifications and equivalents of the present invention, which are made by the contents of the present specification and the accompanying drawings, or directly/indirectly applied to other related technical fields, are included in the scope of the present invention.
Claims (6)
1. The active small-molecule donkey-hide gelatin paste is characterized by being in a viscous liquid paste form, and the solid content is more than 40 Brix. The weight average molecular weight is 600-1500, and the peptide segment is mainly distributed between 180-2000 Da.
2. The preparation method of the active small molecule donkey-hide gelatin paste according to claim 1, which is characterized by comprising the following steps:
s1 pretreatment: processing donkey skin into strips, blocks or granules, soaking and cleaning for later use;
s2 extraction: adding 2-20 times of clear water by weight into the processed donkey skin, and stirring and extracting or refluxing for extraction at the extraction temperature of 70-150 ℃, the stirring speed of 10-60 rpm and the refluxing speed of 1-6T/h. The extraction time is 11 h-73 h;
s3 filtering: extracting until the collagen is completely dissolved, filtering to remove insoluble impurities, and collecting filtrate;
s4 liquid separation: standing the filtrate in an aseptic environment after sterilizing, wherein the standing temperature is 8-68 ℃, and the standing time is 3-24 hours; removing the upper oil layer after layering, and collecting the lower clear liquid;
s5 enzymolysis: adding compound protease into the clear liquid for enzymolysis reaction; the enzymolysis time is 0.4-6.5h, and the reaction temperature is 40-75 ℃; after the reaction is finished, heating to more than 90 ℃ to inactivate enzyme;
s6 concentrating and making into paste: concentrating under reduced pressure at 55-75 deg.C and vacuum degree of-0.001-0.2 Mp until the solid content is above 40Brix, and packaging to obtain the final product.
3. The method for preparing active small molecule donkey-hide gelatin paste according to claim 2, wherein the complex protease comprises metalloprotease and serine protease. One or more of subtilisin, acid protease, alkaline protease, neutral protease, glycosidase, papain, and trypsin can also be contained. The added metal protease and serine protease are 0.1-4% of the dry weight of the raw materials; and the added subtilisin, acid protease, alkaline protease, neutral protease, glycosidase, papain and trypsin are 0-2% of the dry weight of the raw materials.
4. The method for preparing active small molecule donkey-hide gelatin paste according to claim 2, wherein in step S1, 0.01-3% aqueous alkali by weight of clear water can be added to accelerate foaming.
5. The method for preparing active small molecule donkey-hide gelatin paste as claimed in claim 2, wherein the molecular weight of the peptide segment is more than 88% between 180-3000Da, and the molecular weight of the peptide segment is more than 82% between 180-2000 Da.
6. The preparation method of the active small molecule donkey-hide gelatin paste according to claim 2, wherein an auxiliary material is added in the concentration step, and the auxiliary material is one or more of wine, alcohol, sugar and oligosaccharide prebiotics.
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