A donkey-hide gelatin composite polypeptide suitable for postpartum recovery and preparation method thereof

Abstract

The present invention belongs to the technical field of donkey-hide gelatin polypeptides, and specifically relates to a composite bacterial agent and its application in fermenting donkey-hide gelatin to prepare donkey-hide gelatin polypeptides and donkey-hide gelatin paste. The present invention uses a composite bacterial agent composed of Trichoderma CGMCC 3.3711 and yeast CMCC (B) 98007 bacterial liquid to perform specific co-fermentation on traditional Chinese medicine and high-quality donkey-hide gelatin. During the fermentation process, donkey-hide gelatin is decomposed into donkey-hide gelatin polypeptides. When donkey-hide gelatin is decomposed into donkey-hide gelatin polypeptides, multiple effective ingredients in traditional Chinese medicine are introduced to form donkey-hide gelatin composite polypeptides. This method not only has a high donkey-hide gelatin hydrolysis rate, but also has an unexpected effect of promoting milk secretion due to the addition of traditional Chinese medicine. The donkey-hide gelatin composite polypeptide prepared by the present invention has great development prospects in the field of postpartum recovery, and can be used to prepare donkey-hide gelatin polypeptide pastes, etc.

Description

Technical Field

[0001] The invention belongs to the technical field of donkey-hide gelatin polypeptides, and particularly relates to a composite bacterial agent and an application thereof in preparing donkey-hide gelatin polypeptides and donkey-hide gelatin paste by fermenting donkey-hide gelatin. Background Art

[0002] Donkey-hide gelatin is a traditional Chinese medicine with many effects, such as blood tonic, hemostasis, nourishing yin and moistening dryness. Donkey-hide gelatin has significant curative effects and is known as China's intangible cultural heritage for more than 2,000 years. At present, there are many studies on donkey-hide gelatin, and there are in-depth studies on its pharmacological effects and the material basis of its related effects. Donkey-hide gelatin is rich in protein, and it is difficult for patients with indigestion to achieve drug effects in the body. Therefore, it is necessary to enzymatically hydrolyze donkey-hide gelatin to obtain small molecule donkey-hide gelatin polysaccharides. Studies have shown that polypeptides have antioxidant functions and are natural antioxidants.

[0003] Donkey hide gelatin is prepared from donkey skin of equine animals by decoction and concentration, so it has a complex and high content of chemical composition. The main components are composed of 18 kinds of amino acids and 20 kinds of trace elements and small molecular weight collagen hydrolysate. The protein content is rich, up to about 80%. The 18 kinds of amino acids are Lys (2.63%), His (0.59%),

[0004] Ile (0.91%), Arg (4.42%), Thr (3.21%) Met (0.20%), Glu (0.59%), Pro (6.52%), Gly (13.36%), Ala (0.12%), etc. 8 essential amino acids for human body are found in donkey-hide gelatin, accounting for 16.98~21.22% of the total amino acid content. Donkey-hide gelatin also contains a variety of trace metal elements, such as K, Na, Ca, Mg, Fe, Mn, Zn, etc., 8 of which are essential for human body4. Donkey-hide gelatin has many clinical functions such as hemostasis, anti-fatigue, inhibition of tumor growth, improvement of immunity, improvement of gynecological diseases, etc.

[0005] The present invention aims to further expand the application field of donkey-hide gelatin, especially the field of postpartum nourishment. Summary of the invention

[0006] The use of probiotics to modify traditional Chinese medicine is an important direction for the modernization of traditional Chinese medicine. The team of the present invention has also conducted a lot of research on this and unexpectedly discovered a process for preparing composite donkey-hide gelatin peptides that can nourish postpartum mothers, especially promote milk secretion.

[0007] Based on the above reasons, the present invention provides a composite bacterial agent for fermenting donkey-hide gelatin, characterized in that it is composed of Trichoderma CGMCC 3.3711 and yeast CMCC (B) 98007.

[0008] In addition, a method for producing donkey-hide gelatin composite polypeptide suitable for postpartum recovery using the composite bacterial agent is also provided, and the main operating steps are as follows:

[0009] S1. Pretreatment of donkey-hide gelatin: Select high-quality donkey-hide gelatin as raw material, ensure its purity and quality, and crush the donkey-hide gelatin into 400-500 mesh particles for later use;

[0010] S2, Trichoderma CGMCC 3.3711 (purchased from Shanghai Collection Biotechnology Center) bacterial solution preparation: the preparation concentration was 1.5×10 9 ~3×10 9 cfu / ml of Trichoderma CGMCC 3.3711 bacterial solution; yeast CMCC (B) 98007 (purchased from Beijing Biobo Biotechnology Co., Ltd.) bacterial solution preparation: the preparation concentration is 3.5×10 9 ~4.6×10 9 cfu / ml yeast CMCC(B) 98007 bacterial solution for use;

[0011] S3, pretreatment of polygonatum, angelica, wolfberry, dendrobium officinale flower, kudzu root and ginger: polygonatum, angelica, wolfberry, dendrobium officinale flower, kudzu root and ginger are crushed in the same mass ratio and mixed evenly, mixed with water in a mass ratio of 1:1.5, and then added with the Trichoderma liquid and yeast liquid prepared in step S2 and fermented in a tank, wherein the mass ratio of the Chinese medicine mixture: Trichoderma liquid: yeast liquid is 1-4:0.5-2:0.5-2, the fermentation temperature is 32-35°C, and the fermentation time is 12-24 hours; the step of using the Trichoderma liquid and the yeast liquid to ferment the Chinese medicine is to enhance the effect of the Chinese medicine and obtain more beneficial substances in the Chinese medicine;

[0012] S4, after the fermentation is completed, the supernatant is taken, and then the crushed donkey-hide gelatin in step S1 is added, and the mixture is stirred evenly at a temperature of 32-35°C, and then the temperature is maintained at 32-35°C for fermentation for 6-12 hours to obtain donkey-hide gelatin polypeptide liquid containing traditional Chinese medicine fermentation liquid, wherein the fermentation liquid and donkey-hide gelatin are added at a mass ratio of 1:1; in this step, the traditional Chinese medicine fermentation liquid is used to ferment donkey-hide gelatin, and the purpose is to obtain donkey-hide gelatin polypeptides by decomposing donkey-hide gelatin through microorganisms;

[0013] S5. After fermentation is completed, 3% H2O2 solution is added to inactivate Trichoderma CGMCC 3.3711 and yeast CMCC(B) 98007, and then the donkey-hide gelatin polypeptide liquid containing the traditional Chinese medicine fermentation liquid is filtered to remove solid particles and impurities, and then an appropriate amount of 2mM NaOH is added to induce the precipitation of donkey-hide gelatin polypeptide, and then centrifugation is performed to collect the precipitate, and the collected precipitate is dried at 30-40°C to obtain the donkey-hide gelatin composite polypeptide for postpartum recovery.

[0014] Preferably, high-quality donkey-hide gelatin is selected in step S1 of the present invention.

[0015] Preferably, the concentration of the Trichoderma CGMCC 3.3711 bacterial solution used in step S2 of the present invention is 2.2×10 9 cfu / ml.

[0016] Preferably: The concentration of yeast CMCC (B) 98007 used in step S2 of the present invention is 3.9×10 9 cfu / ml.

[0017] Preferably: in step S3 of the present invention, polygonatum sibiricum, angelica sinensis, wolfberry, dendrobium officinale flower, kudzu root and ginger are crushed in the same mass ratio and then mixed evenly.

[0018] Preferably: in step S3 of the present invention, the Chinese medicine mixture: Trichoderma liquid: yeast liquid are evenly mixed in a mass ratio of 1:0.5:0.5.

[0019] Preferably, the supernatant in step S4 of the present invention is not extinguished, but is directly mixed with the melted donkey-hide gelatin in step S1 for fermentation.

[0020] Preferably: the donkey-hide gelatin composite polypeptide obtained in step S5 of the present invention is used to prepare the donkey-hide gelatin composite polypeptide paste;

[0021] The specific preparation method is as follows:

[0022] Heat a high-quality donkey-hide gelatin block at 80-90°C and melt it, and keep the temperature at 60-70°C to maintain the melted state after melting, add the donkey-hide gelatin composite polypeptide prepared by the present invention to the high-quality donkey-hide gelatin at a mass ratio of 1:0.5, and then add black sesame, wolfberry, red dates and walnuts to the high-quality donkey-hide gelatin at a mass ratio of 1:0.5:0.2:0.2:0.1 respectively, mix and stir evenly, flatten and cool, and then cut into small cubes with a length of 6-8cm, a width of 3-4cm, and a thickness of 1-1.5cm to obtain donkey-hide gelatin polypeptide paste.

[0023] The present invention has the following beneficial effects:

[0024] 1. After a large number of experiments, our team unexpectedly discovered that the use of specific Trichoderma and yeast to ferment a specific compound donkey-hide gelatin formula can remove unnecessary ingredients or enhance the efficacy, and fully extract the beneficial substances in traditional Chinese medicine.

[0025] 2. Mixing high-quality donkey-hide gelatin with traditional Chinese medicine fermentation liquid for co-fermentation can effectively obtain donkey-hide gelatin polypeptides.

[0026] 3. The donkey-hide gelatin prepared by the present invention has an unexpected effect of promoting milk secretion.

[0027] 4. The donkey-hide gelatin composite polypeptide prepared by the present invention can be used to prepare donkey-hide gelatin polypeptide ointment, oral liquid, decoction and tablets, etc., and is very convenient to use.

[0028] 5. Polygonatum: Polygonatum can replenish qi and nourish yin, strengthen the spleen, moisten the lungs, and benefit the kidneys. Angelica: Angelica is widely used in traditional Chinese medicine and is believed to regulate the female reproductive system, promote uterine contraction and recovery, and help restore menstruation and uterine health. Lycium barbarum: Lycium barbarum contains antioxidants and multiple vitamins, which help strengthen the immune system and relieve postpartum fatigue. Dendrobium officinale flower: Dendrobium officinale flower can refresh and relieve depression, help relieve mental tension, keep the mind clear, relieve mental depression, and prevent postpartum depression. Pueraria root: Pueraria root is believed to have the effects of nourishing qi and blood, nourishing yin and clearing heat. Ginger: Ginger is believed to have anti-inflammatory and analgesic effects, which can prevent wound inflammation and promote wound healing. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 This is a comparison chart of the hydrolysis rates of donkey-hide gelatin of Example 1, Comparative Example 1 and Comparative Example 2 of the present invention.

[0030] Figure 2 It is a comparison chart of the hydrolysis rate of donkey-hide gelatin in Example 2 of the present invention and Comparative Examples 3-5. DETAILED DESCRIPTION

[0031] The following describes the embodiments of the present invention by specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention. It should be noted that the following embodiments and features in the embodiments can be combined with each other without conflict.

[0032] Example 1

[0033] S1. Pretreatment of donkey-hide gelatin: Select high-quality donkey-hide gelatin as raw material, ensure its purity and quality, and crush the donkey-hide gelatin into 400-500 mesh particles for later use;

[0034] S2. Preparation of Trichoderma CGMCC 3.3711 bacterial solution: the concentration was 1.5×10 9 cfu / ml for the preparation of Trichoderma CGMCC3.3711 bacterial solution; for the preparation of yeast CMCC(B) 98007 bacterial solution: the preparation concentration is 3.5×10 9 cfu / ml yeast CMCC(B) 98007 bacterial solution for use;

[0035] S3, pretreatment of polygonatum, angelica, wolfberry, dendrobium officinale flower, kudzu root and ginger: polygonatum, angelica, wolfberry, dendrobium officinale flower, kudzu root and ginger are crushed in the same mass ratio and mixed evenly, mixed with water in a mass ratio of 1:1.5, and then added with the Trichoderma liquid and yeast liquid prepared in step S2, and fermented in a tank, wherein the mass ratio of the Chinese medicine mixture: Trichoderma liquid: yeast liquid is 1:0.5:0.5, the fermentation temperature is 32°C, and the fermentation time is 12 hours;

[0036] S4, after the fermentation is completed, filter, take the supernatant after filtration, then pour into the melted donkey-hide gelatin in step S1, stir evenly at a temperature of 32°C, and then maintain the temperature at 32°C for 6-12 hours to obtain donkey-hide gelatin polypeptide liquid containing traditional Chinese medicine fermentation liquid, wherein the fermentation liquid: donkey-hide gelatin are added at a mass ratio of 1:1;

[0037] S5. After the fermentation is completed, 3% H2O2 solution is added to inactivate Trichoderma CGMCC 3.3711 and yeast CMCC(B) 98007, and then the donkey-hide gelatin polypeptide liquid containing the traditional Chinese medicine fermentation liquid is filtered to remove solid particles and impurities, and then an appropriate amount of 2mM NaOH is added to induce the precipitation of the donkey-hide gelatin polypeptide, and then centrifugation is performed to collect the precipitate, and the collected precipitate is dried at 30°C to obtain the donkey-hide gelatin composite polypeptide for postpartum recovery.

[0038] Comparative Example 1: Except that the Trichoderma CGMCC 3.3711 bacterial solution was not added in step S3, the remaining steps were the same as those in Example 1.

[0039] Comparative Example 2: Except that yeast CMCC(B) 98007 solution was not added in step S3, the remaining steps were the same as those in Example 1.

[0040] Figure 1 The figure is a comparison chart of the hydrolysis rate of donkey-hide gelatin of Example 1, Comparative Example 1 and Comparative Example 2 of the present invention. The present invention directly mixes the Chinese medicine fermentation liquid without fungus inactivation with high-quality donkey-hide gelatin, and uses Trichoderma CGMCC 3.3711 and yeast CMCC (B) 98007 in the Chinese medicine hair spray liquid to co-ferment the Chinese medicine fermentation liquid and high-quality donkey-hide gelatin. During the fermentation, the donkey-hide gelatin is decomposed into donkey-hide gelatin polypeptides under the action of Trichoderma CGMCC3.3711 and yeast CMCC (B) 98007. Figure 1 It can be seen that the hydrolysis degree of donkey-hide gelatin was significantly reduced without the use of Trichoderma CGMCC 3.3711 culture liquid or yeast CMCC (B) 98007 culture liquid, which indicates that the significant synergistic effect between Trichoderma CGMCC 3.3711 and yeast CMCC (B) 98007 can promote the hydrolysis of donkey-hide gelatin and thus obtain more donkey-hide gelatin peptides.

[0041] Example 2

[0042] S1. Pretreatment of donkey-hide gelatin: Select high-quality donkey-hide gelatin as raw material, ensure its purity and quality, and crush the donkey-hide gelatin into 400-500 mesh particles for later use;

[0043] S2. Preparation of Trichoderma CGMCC 3.3711 bacterial solution: the preparation concentration is 3×10 9 cfu / ml for the preparation of Trichoderma CGMCC 3.3711 bacterial solution; for the preparation of yeast CMCC (B) 98007 bacterial solution: the preparation concentration is 4.6×10 9 cfu / ml yeast CMCC(B) 98007 bacterial solution for use;

[0044] S3, pretreatment of polygonatum, angelica, wolfberry, dendrobium officinale flower, kudzu root and ginger: polygonatum, angelica, wolfberry, dendrobium officinale flower, kudzu root and ginger are crushed in the same mass ratio and mixed evenly, mixed with water in a mass ratio of 1:1.5, and then added with the Trichoderma liquid and yeast liquid prepared in step S2, and fermented in a tank, wherein the mass ratio of the Chinese medicine mixture: Trichoderma liquid: yeast liquid is 2:1:1, the fermentation temperature is 35°C, and the fermentation time is 24 hours;

[0045] S4, after the fermentation is completed, filter, take the supernatant after filtration, then pour into the donkey-hide gelatin melted in step S1, stir evenly at a temperature of 35°C, and then maintain the temperature at 35°C for 12 hours to obtain donkey-hide gelatin polypeptide liquid containing traditional Chinese medicine fermentation liquid, wherein the fermentation liquid: donkey-hide gelatin are added at a mass ratio of 1:1;

[0046] S5. After fermentation is completed, 3% H2O2 solution is added to inactivate Trichoderma CGMCC 3.3711 and yeast CMCC(B) 98007, and then the donkey-hide gelatin polypeptide liquid containing the traditional Chinese medicine fermentation liquid is filtered to remove solid particles and impurities, and then an appropriate amount of 2mM NaOH is added to induce the precipitation of donkey-hide gelatin polypeptide, and then centrifugation is performed to collect the precipitate, and the collected precipitate is dried at 30-40°C to obtain the donkey-hide gelatin composite polypeptide for postpartum recovery.

[0047] Comparative Example 3: Except that the Chinese medicine fermentation liquid is replaced with hot water stirring in step S4, the remaining steps are the same as those in Example 2.

[0048] Comparative Example 4: Except that the Trichoderma CGMCC 3.3711 in step S2 was replaced by Trichoderma CGMCC3.2942 (purchased from Beijing Biobo Biotechnology Co., Ltd.), the remaining steps were the same as those in Example 2.

[0049] Comparative Example 5: Except that the yeast CMCC (B) 98007 in step S2 was replaced by yeast ATCC 9763 (purchased from Ruichu Biotechnology (Jiangsu) Co., Ltd.), the remaining steps were the same as those in Example 2.

[0050] Figure 2 This is a comparison chart of the hydrolysis rates of donkey-hide gelatin in Example 2 of the present invention and Comparative Examples 3-5. Figure 2 It can be seen that the decomposition rate of Example 2 using the fermentation decomposition method can reach 67%, and the hydrolysis rate using the hot water stirring hydrolysis method is significantly reduced, showing the superiority of fungal decomposition. According to the results of Comparative Examples 4 and 3, it can be seen that Trichoderma CGMCC 3.3711 and yeast CMCC (B) 98007 are specific for decomposing donkey-hide gelatin, and the synergistic effect between the two bacteria is also specific.

[0051] Example 3

[0052] S1. Pretreatment of donkey-hide gelatin: Select high-quality donkey-hide gelatin as raw material, ensure its purity and quality, and crush the donkey-hide gelatin into 400-500 mesh particles for later use;

[0053] S2. Preparation of Trichoderma CGMCC 3.3711 bacterial solution: the preparation concentration is 2.2×10 9 cfu / ml for the preparation of Trichoderma CGMCC3.3711 bacterial solution; for the preparation of yeast CMCC(B) 98007 bacterial solution: the preparation concentration is 3.9×10 9 cfu / ml yeast ATCC 12845 bacterial solution is reserved;

[0054] S3, pretreatment of polygonatum, angelica, wolfberry, dendrobium officinale flower, kudzu root and ginger: polygonatum, angelica, wolfberry, dendrobium officinale flower, kudzu root and ginger are crushed in the same mass ratio and mixed evenly, mixed with water in a mass ratio of 1:1.5, and then added with the Trichoderma liquid and yeast liquid prepared in step S2, and fermented in a tank, wherein the mass ratio of the Chinese medicine mixture: Trichoderma liquid: yeast liquid is 1:0.5:0.5, the fermentation temperature is 33°C, and the fermentation time is 18 hours;

[0055] S4, after the fermentation is completed, filter, take the supernatant after filtration, then pour into the donkey-hide gelatin melted in step S1, stir evenly at a temperature of 33°C, and then maintain the temperature at 32-35°C for 6-12 hours to obtain donkey-hide gelatin polypeptide liquid containing traditional Chinese medicine fermentation liquid, wherein the fermentation liquid: donkey-hide gelatin are added at a mass ratio of 1:1;

[0056] S5. After fermentation is completed, 3% H2O2 solution is added to inactivate Trichoderma CGMCC 3.3711 and yeast CMCC(B) 98007, and then the donkey-hide gelatin polypeptide liquid containing the traditional Chinese medicine fermentation liquid is filtered to remove solid particles and impurities, and then an appropriate amount of 2mM NaOH is added to induce the precipitation of donkey-hide gelatin polypeptide, and then centrifugation is performed to collect the precipitate, and the collected precipitate is dried at 35°C to obtain the donkey-hide gelatin composite polypeptide for postpartum recovery.

[0057] Comparative Example 6: Except that no Chinese medicine is added for fermentation in step S3, the remaining steps are the same as those in Example 3.

[0058] Comparative Example 7: Except that Pueraria root is not added in step S4, the remaining steps are the same as those in Example 3.

[0059] Comparative Example 8: Except that ginger is not added in step S4, the remaining steps are the same as those in Example 3.

[0060] Example 4

[0061] Animal experiments

[0062] Lactation conditioning experiment

[0063] Animal grouping, modeling and drug administration

[0064] 70 SPF-grade healthy KM pregnant mice (supplied by Beijing Biotechnology Co., Ltd., animal production license SCXK (Beijing) 2016-0002), 130 days old, weighing 92±11g, were raised in our company's SPF-grade animal breeding room with relatively constant temperature (25℃), constant humidity (75%), fed with conventional mouse pellet feed, and allowed free access to food and water.

[0065] 55 female mice with a time difference of no more than 24 hours before and after the birth were selected as experimental research subjects, and the number of pups in each litter was adjusted to 5. The female mice were randomly divided into 11 groups, namely, the insufficiency model group, the normal group, the positive control estradiol group, the example group (Example 3), and the comparative example groups (Comparative Example 1, Comparative Example 2, Comparative Example 4, Comparative Example 5, Comparative Example 6, Comparative Example 7 and Comparative Example 8), with 5 mice in each group, using a random number table method.

[0066] The day of delivery was the 0th day. From the 3rd day after delivery, except for the normal group, the other groups were gavaged with bromocriptine (1.6 mg / kg) daily to establish a postpartum milk deficiency model in mice for 10 days. At the same time as the modeling, according to the human and animal dose conversion method, the example group (Example 3), the comparative example group (Comparative Example 1, Comparative Example 2, Comparative Example 4, Comparative Example 5, Comparative Example 6, Comparative Example 7 and Comparative Example 8) were fed with 1.2g / 100g·d of the donkey-hide gelatin composite polypeptide prepared in Example 3, Comparative Example 1, Comparative Example 2, Comparative Example 4, Comparative Example 5, Comparative Example 6, Comparative Example 7 and Comparative Example 8, respectively, the estradiol group was given 17α-estradiol (1 g / kg), and the normal group and the milk deficiency model group were given an equal amount of physiological saline. After gavage for 10 days (the 13th day after delivery), the mice were killed, and the blood and mammary tissue of the mother mice were taken for index detection. The litter weight gain of pups was measured, and the milk production and PRL were tested with reference to the methods of the prior art.

[0067] (3) Index detection

[0068] Measurement of the increase in the weight of the litter of pups: The increase in the weight of the litter of pups was obtained by subtracting the first litter weight from the last litter weight after the 10th day.

[0069] Measurement of milk production: Starting from the third day of administration, all litters of pups were weighed at a fixed time every day (W1), and the pups were weighed again after being isolated from the mother for 4 hours (W2), and then weighed for the third time after being placed in a cage with the mother for lactation for 1 hour (W3). The hourly milk production of the mother was calculated by the weight difference of all the pups before and after lactation for 1 hour (W3-W2), plus the average basal metabolic rate of all the pups in the litter for 1 hour ((W1-W2) / 4). That is, hourly milk production = W3-W2+(W1-W2) / 4.

[0070] The results of the weight increase of the litter of young mice are shown in Table 1:

[0071] Table 1

[0072] The milk yield (mL) is shown in Table 2:

[0073]

[0074] Table 2

[0075]

[0076] According to the results of the increase in the litter weight and the milk production of the lactation model group in Table 1 and Table 2, the lactation model was successfully constructed. According to the results of Example 3, Comparative Examples 1 and 2, when Trichoderma CGMCC 3.3711 or yeast CMCC (B) 98007 was not used for fermentation, the weight gain of the litter weight of the young mice and the milk production of the mother mice were significantly reduced, indicating that the synergistic effect between Trichoderma CGMCC 3.3711 and yeast CMCC (B) 98007 can promote milk secretion. According to Example 3, Comparative Examples 4 and Comparative Examples 5, Trichoderma CGMCC 3.3711 and yeast CMCC (B) 98007 are specific to promoting milk fermentation. According to Example 3, Comparative Example 6, Comparative Example 7 and Comparative Example 8, the addition of Chinese herbal fermentation products has an unexpected effect on promoting milk secretion, and among these 6 medicines, the simultaneous addition of Pueraria root and ginger will produce a synergistic effect, wherein Pueraria root can promote milk secretion, and ginger can drive blood circulation and accelerate milk secretion.

[0077] The above-mentioned embodiments only express the specific implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the protection scope of the present invention. It should be pointed out that, for those skilled in the art, without departing from the concept of the present invention, any other changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principle of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A method for preparing a composite donkey-hide gelatin polypeptide for postpartum recovery, characterized in that: The specific method is as follows: S1. Pretreatment of donkey-hide gelatin: crush the donkey-hide gelatin for standby use; S2, Trichoderma reesei ( Trichoderma reesei ) Preparation of CGMCC 3.3711 bacterial solution: the concentration is 1.5×10 9 ~3×10 9 cfu / ml of Trichoderma reesei CGMCC 3.3711 bacterial solution; Saccharomyces cerevisiae ( Saccharomyces cerevisiae )CMCC(B) 98007 bacterial solution preparation: the preparation concentration is 3.5×10 9 ~4.6×10 9 cfu / ml of brewer's yeast CMCC(B) 98007 bacterial solution for use; S3, pretreatment of polygonatum, angelica, wolfberry, dendrobium officinale flower, kudzu root and ginger: crush polygonatum, angelica, wolfberry, dendrobium officinale flower, kudzu root and ginger and mix them evenly, mix them with water at a mass ratio of 1:1.5, then add the Trichoderma liquid and yeast liquid prepared in step S2, and ferment them in a tank, the fermentation temperature is 32-35°C, and the fermentation time is 12-24 hours; S4, after the fermentation is completed, filter, take the supernatant after filtration, then add the crushed donkey-hide gelatin in step S1, stir evenly at a temperature of 32-35°C, and then maintain the temperature at 32-35°C for 6-12 hours to obtain donkey-hide gelatin polypeptide liquid containing traditional Chinese medicine fermentation liquid, wherein the fermentation liquid: donkey-hide gelatin are added at a mass ratio of 1:1; S5. After fermentation is completed, 3% H2O2 solution is added to inactivate Trichoderma CGMCC 3.3711 and yeast CMCC(B) 98007, and then the donkey-hide gelatin polypeptide liquid containing the traditional Chinese medicine fermentation liquid is filtered to remove solid particles and impurities, and then an appropriate amount of 2mM NaOH is added to induce the precipitation of donkey-hide gelatin polypeptide, and then centrifugation is performed to collect the precipitate, and the collected precipitate is dried at 30-40°C to obtain the donkey-hide gelatin composite polypeptide for postpartum recovery.

2. The method for preparing the composite donkey-hide gelatin polypeptide for postpartum recovery according to claim 1, characterized in that: The concentration of Trichoderma reesei CGMCC 3.3711 bacterial solution used in step S2 is 2.2×10 9 cfu / ml.

3. The method for preparing the composite donkey-hide gelatin polypeptide for postpartum recovery according to claim 1, characterized in that: The concentration of the brewer's yeast CMCC (B) 98007 culture solution used in step S2 is 3.9×10 9 cfu / ml.

4. The method for preparing the composite donkey-hide gelatin polypeptide for postpartum recovery according to claim 3, characterized in that: In step S3, polygonatum sibiricum, angelica sinensis, wolfberry fruit, dendrobium officinale flower, kudzu root and ginger are crushed in the same mass ratio and then mixed evenly.

5. The method for preparing the composite donkey-hide gelatin polypeptide for postpartum recovery according to claim 4, characterized in that: In the step S3, the Chinese medicine mixture: the Trichoderma liquid: the yeast liquid are evenly mixed in a mass ratio of 1:0.5:0.

5.

6. The method for preparing the composite donkey-hide gelatin polypeptide for postpartum recovery according to claim 5, characterized in that: The supernatant in step S4 is not inactivated, but is directly mixed with the melted donkey-hide gelatin in step S1 for fermentation.

7. The donkey-hide gelatin composite polypeptide prepared by the preparation method according to any one of claims 1 to 6.

8. A method for preparing donkey-hide gelatin paste, characterized in that: The donkey-hide gelatin composite polypeptide according to claim 7 is used to prepare donkey-hide gelatin composite polypeptide paste; The specific preparation method is as follows: Heat the high-quality donkey-hide gelatin block at 80-90℃ and melt it. After melting, keep the temperature at 60-70℃ to maintain the melted state. Mix it with high-quality donkey-hide gelatin in a mass ratio of 1: 0.5 add the donkey-hide gelatin composite polypeptide as claimed in claim 7, and then add black sesame, wolfberry, red dates and walnuts in a mass ratio of 1:0.5:0.2:0.2:0.1 to the high-quality donkey-hide gelatin, mix and stir evenly, spread flat and cool, and then cut into small cubes of 6-8 cm in length, 3-4 cm in width, and 1-1.5 cm in thickness to obtain donkey-hide gelatin polypeptide paste.

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