CN117050888A - Composite microbial inoculum and application thereof in preparation of donkey-hide gelatin polypeptide and donkey-hide gelatin paste by fermenting donkey-hide gelatin - Google Patents
Composite microbial inoculum and application thereof in preparation of donkey-hide gelatin polypeptide and donkey-hide gelatin paste by fermenting donkey-hide gelatin Download PDFInfo
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- CN117050888A CN117050888A CN202311293889.2A CN202311293889A CN117050888A CN 117050888 A CN117050888 A CN 117050888A CN 202311293889 A CN202311293889 A CN 202311293889A CN 117050888 A CN117050888 A CN 117050888A
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Abstract
The invention belongs to the technical field of donkey-hide gelatin polypeptides, and particularly relates to a compound microbial inoculum and application thereof in preparation of donkey-hide gelatin polypeptides and donkey-hide gelatin paste by fermenting donkey-hide gelatin. The invention uses the compound microbial inoculum composed of trichoderma CGMCC 3.3711 and saccharomycete CMCC (B) 98007 to carry out specific co-fermentation on traditional Chinese medicine and high-quality donkey-hide gelatin, the donkey-hide gelatin is decomposed into donkey-hide gelatin polypeptide in the fermentation process, and various active ingredients in the traditional Chinese medicine are introduced while the donkey-hide gelatin is decomposed into the donkey-hide gelatin polypeptide, thus forming the donkey-hide gelatin compound polypeptide. The method has high donkey-hide gelatin hydrolysis rate, and unexpected effect of promoting milk secretion due to the addition of traditional Chinese medicine. The donkey-hide gelatin composite polypeptide prepared by the invention has great development prospect in the field of postpartum recovery, and can be used for preparing donkey-hide gelatin polypeptide paste and the like.
Description
Technical Field
The invention belongs to the technical field of donkey-hide gelatin polypeptides, and particularly relates to a compound microbial inoculum and application thereof in preparation of donkey-hide gelatin polypeptides and donkey-hide gelatin paste by fermenting donkey-hide gelatin.
Background
Donkey-hide gelatin is a traditional Chinese medicinal material and has many effects such as replenishing blood, stopping bleeding, nourishing yin, moistening dryness and the like. Donkey-hide gelatin has remarkable curative effect and is praised as a non-material cultural heritage in China over 2000. At present, donkey-hide gelatin has more researches and has more intensive researches on pharmacological actions and substance bases of related actions. Donkey-hide gelatin has rich protein content, and is difficult to achieve in vivo efficacy for patients with indigestion. Therefore, the donkey-hide gelatin needs to be subjected to enzymolysis to obtain small-molecule donkey-hide gelatin polynnaeus. Researches prove that the polypeptide has an antioxidant function and is a natural antioxidant.
Donkey-hide gelatin is prepared from donkey skin of equine animals by water decoction and concentration, and thus has a complex and high content of chemical components. The main component consists of 18 amino acids and 20 microelements and is collagen hydrolysate with small molecular weight. The protein content is rich and reaches about 80 percent. 18. The amino acids are Lys (2.63%), his (0.59%)
Ile (0.91%), arg (4.42%), thr (3.21%), met (0.20%), glu (0.59%), pro (6.52%), gly (13.36%), ala (0.12%), and the like. 8 human essential amino acids are found in donkey-hide gelatin, wherein the amino acids account for 16.98-21.22% of the total amino acid content. Donkey-hide gelatin also contains various trace metal elements such as K, na, ca, mg, fe, mn, zn, etc., 8 of which are 4 necessary for human body. Colla Corii Asini has hemostatic, antifatigue, tumor growth inhibiting, immunity enhancing, and gynecological diseases improving effects.
The invention aims to further expand the application field of donkey-hide gelatin, in particular to the postpartum nourishing field.
Disclosure of Invention
The probiotics are an important direction of traditional Chinese medicine modernization, and the invention team also carries out a great deal of research on the probiotics, and the preparation process of the compound donkey-hide gelatin peptide which can prepare postpartum nourishing, especially promote milk secretion is unexpectedly discovered.
Based on the reasons, the invention provides a composite microbial inoculum for fermenting donkey-hide gelatin, which is characterized in that: consists of Trichoderma CGMCC 3.3711 and saccharomycete CMCC (B) 98007.
In addition, the method for producing the donkey-hide gelatin composite polypeptide suitable for postpartum recovery by using the composite microbial inoculum is provided, and the main operation steps are as follows:
s1, pretreatment of donkey-hide gelatin: selecting high-quality donkey-hide gelatin as a raw material, ensuring the purity and quality of the donkey-hide gelatin, and crushing the donkey-hide gelatin into 400-500 mesh particles for later use;
s2, preparing Trichoderma CGMCC 3.3711 (purchased from Shanghai collection biotechnology center) bacterial liquid: preparation of 1.5X10 concentration 9 ~3×10 9 Preparing cfu/ml trichoderma CGMCC 3.3711 bacterial liquid; preparation of a Yeast CMCC (B) 98007 (from Beijing Bai Ou Bo Wei Biotechnology Co., ltd.) bacterial liquid: preparation of the concentration was 3.5X10 9 ~4.6×10 9 cfu/ml of saccharomycete CMCC (B) 98007 bacterial liquid for standby;
s3, pretreatment of rhizoma polygonati, chinese angelica, medlar, dendrobium candidum flowers, kudzuvine roots and ginger: pulverizing rhizoma polygonati, chinese angelica, medlar, dendrobium officinale flower, radix puerariae and ginger according to the same mass ratio, uniformly mixing, and mixing with water according to the mass ratio of 1:1.5, mixing, then adding the trichoderma liquid and the saccharomycete liquid prepared in the step S2, and carrying out jar fermentation, wherein the traditional Chinese medicine mixture is as follows: trichoderma liquid: the mass ratio of the saccharomycete liquid is 1-4: 0.5-2: 0.5-2, wherein the fermentation temperature is 32-35 ℃ and the fermentation time is 12-24 hours; the step uses trichoderma liquid and saccharomycete liquid to ferment the traditional Chinese medicine, so as to enhance the effect of the traditional Chinese medicine and obtain more beneficial substances in the traditional Chinese medicine;
s4, filtering after fermentation, taking supernatant, adding the donkey-hide gelatin crushed in the step S1, stirring uniformly at the temperature of 32-35 ℃, and fermenting for 6-12 hours at the temperature of 32-35 ℃ to obtain donkey-hide gelatin polypeptide liquid containing traditional Chinese medicine fermentation liquid, wherein the fermentation liquid is prepared by the steps of: the mass ratio of the donkey-hide gelatin is 1:1, adding; the step of fermenting donkey-hide gelatin by using a traditional Chinese medicine fermentation liquid, and aims to obtain donkey-hide gelatin polypeptide by decomposing donkey-hide gelatin by microorganisms;
s5, after fermentation is completed, adding 3%H 2 O 2 Inactivating Trichoderma CGMCC 3.3711 and yeast CMCC (B) 98007, filtering the donkey-hide gelatin polypeptide liquid containing the Chinese medicinal fermentation liquid to remove solid particles and impurities, adding 2mM NaOH to precipitate donkey-hide gelatin polypeptide, centrifuging to collect precipitate, and drying at 30-40deg.CDrying to obtain colla Corii Asini compound polypeptide for postpartum recovery.
Preferably: in the step S1, high-quality donkey-hide gelatin is selected.
Preferably: the concentration of the trichoderma CGMCC 3.3711 bacterial liquid used in the step S2 is 2.2 multiplied by 10 9 cfu/ml。
Preferably: the concentration of the yeast CMCC (B) 98007 used in the step S2 is 3.9X10 9 cfu/ml。
Preferably: in the invention, in the step S3, the rhizoma polygonati, the Chinese angelica, the medlar, the dendrobium officinale flower, the kudzuvine root and the ginger are crushed according to the same mass ratio and then are uniformly mixed.
Preferably: in the step S3, the traditional Chinese medicine mixture is as follows: trichoderma liquid: the mass ratio of the saccharomycete liquid is 1:0.5:0.5, mixing evenly.
Preferably: the supernatant in the step S4 is not subjected to fire extinguishment, and is directly mixed with the melted donkey-hide gelatin in the step S1 for fermentation.
Preferably: the donkey-hide gelatin composite polypeptide obtained in the step S5 is used for preparing donkey-hide gelatin composite polypeptide paste;
the preparation method comprises the following steps:
heating the high-quality donkey-hide gelatin block at 80-90 ℃ and then melting, keeping the temperature at 60-70 ℃ after melting, and keeping a melting state, wherein the mass ratio of the donkey-hide gelatin block to the high-quality donkey-hide gelatin block is 1:0.5 adding the donkey-hide gelatin composite polypeptide prepared by the invention, and then mixing the donkey-hide gelatin composite polypeptide with high-quality donkey-hide gelatin according to a mass ratio of 1:0.5:0.2:0.2: and 0.1, respectively adding black sesame, medlar, red date and walnut, uniformly mixing and stirring, spreading, airing and cooling, and cutting into small blocks with the length of 6-8cm, the width of 3-4cm and the thickness of 1-1.5cm to obtain the donkey-hide gelatin polypeptide paste.
The invention has the following beneficial effects:
1. through a large number of experiments, the team unexpectedly finds that the specific compound donkey-hide gelatin formula is fermented by adopting specific trichoderma and saccharomycetes so as to remove unnecessary components or enhance the drug effect, and the beneficial substances in the traditional Chinese medicine are fully extracted.
2. The donkey-hide gelatin polypeptide can be effectively obtained by mixing and co-fermenting high-quality donkey-hide gelatin and Chinese medicinal fermentation broth.
3. The donkey-hide gelatin prepared by the invention has unexpected effect of promoting milk secretion.
4. The donkey-hide gelatin composite polypeptide prepared by the invention can be used for preparing donkey-hide gelatin polypeptide paste, oral liquid, decoction, tablets and the like, and is very convenient to use.
5. Rhizoma polygonati: polygonatum sibiricum has the effects of invigorating qi, nourishing yin, strengthening spleen, moistening lung and tonifying kidney. Chinese angelica root: chinese angelica is widely used in traditional Chinese medicine, is considered to be capable of conditioning female reproductive system, promoting uterine contraction and recovery, and is beneficial to recovery of menstruation and health of uterus. Wolfberry fruit: fructus Lycii contains antioxidant and multivitamins, and helps to enhance immune system and relieve puerperal fatigue. Flower of Dendrobium officinale: dendrobium officinale flower can refresh and resolve depression, is helpful for relieving mental stress, keeping consciousness, relieving mental depression, and can prevent postpartum depression. Radix Puerariae: the kudzuvine root is considered to have the effects of nourishing qi and blood, nourishing yin and clearing heat. Ginger: ginger is considered to have anti-inflammatory and analgesic effects, and can prevent inflammation of wounds, thereby facilitating wound healing.
Drawings
FIG. 1 is a graph showing the comparison of the hydrolysis rates of donkey-hide gelatin of comparative example 1 and comparative example 2, according to the present invention.
FIG. 2 is a graph showing the comparison of the hydrolysis rate of colla Corii Asini in example 2 and comparative examples 3-5 according to the present invention.
Detailed Description
Other advantages and effects of the present invention will become apparent to those skilled in the art from the following disclosure, which describes the embodiments of the present invention with reference to specific examples. The invention may be practiced or carried out in other embodiments that depart from the specific details, and the details of the present description may be modified or varied from the spirit and scope of the present invention. It should be noted that the following embodiments and features in the embodiments may be combined with each other without conflict.
Example 1
S1, pretreatment of donkey-hide gelatin: selecting high-quality donkey-hide gelatin as a raw material, ensuring the purity and quality of the donkey-hide gelatin, and crushing the donkey-hide gelatin into 400-500 mesh particles for later use;
s2, preparing Trichoderma CGMCC 3.3711 bacterial liquid: preparation of 1.5X10 concentration 9 Preparing cfu/ml trichoderma CGMCC 3.3711 bacterial liquid; preparation of saccharomycete CMCC (B) 98007 bacterial liquid: preparation of the concentration was 3.5X10 9 cfu/ml of saccharomycete CMCC (B) 98007 bacterial liquid for standby;
s3, pretreatment of rhizoma polygonati, chinese angelica, medlar, dendrobium candidum flowers, kudzuvine roots and ginger: pulverizing rhizoma polygonati, chinese angelica, medlar, dendrobium officinale flower, radix puerariae and ginger according to the same mass ratio, uniformly mixing, and mixing with water according to the mass ratio of 1:1.5, mixing, then adding the trichoderma liquid and the saccharomycete liquid prepared in the step S2, and carrying out jar fermentation, wherein the traditional Chinese medicine mixture is as follows: trichoderma liquid: the mass ratio of the saccharomycete liquid is 1:0.5:0.5, the fermentation temperature is 32 ℃, and the fermentation time is 12 hours;
s4, filtering after fermentation, taking supernatant, pouring the supernatant into the donkey-hide gelatin melted in the step S1, uniformly stirring at the temperature of 32 ℃, and then keeping the temperature of 32 ℃ for fermentation for 6-12 hours to obtain donkey-hide gelatin polypeptide liquid containing traditional Chinese medicine fermentation liquid, wherein the fermentation liquid is prepared by the steps of: the mass ratio of the donkey-hide gelatin is 1:1, adding;
s5, after fermentation is completed, adding 3%H 2 O 2 Inactivating Trichoderma CGMCC 3.3711 and saccharomycete CMCC (B) 98007, filtering the donkey-hide gelatin polypeptide liquid containing the Chinese medicine fermentation liquid to eliminate solid grains and impurity, adding 2mM NaOH to precipitate donkey-hide gelatin polypeptide, centrifuging to obtain precipitate, and drying the precipitate at 30 deg.c to obtain the composite donkey-hide gelatin polypeptide for post partum recovery.
Comparative example 1: the steps are the same as in example 1 except that the Trichoderma CGMCC 3.3711 bacterial liquid is not added in the step S3.
Comparative example 2: the procedure of example 1 was repeated except that the bacterial liquid of yeast CMCC (B) 98007 was not added in step S3.
FIG. 1 is a graph showing the comparison of the hydrolysis rates of donkey-hide gelatin of comparative example 1 and comparative example 2, according to the present invention. According to the invention, the traditional Chinese medicine fermentation liquid which is not subjected to fungus inactivation is directly mixed with the high-quality donkey-hide gelatin, the traditional Chinese medicine fermentation liquid and the high-quality donkey-hide gelatin are co-fermented by using the trichoderma CGMCC 3.3711 and the saccharomycete CMCC (B) 98007 in the traditional Chinese medicine fermentation liquid, and the donkey-hide gelatin is decomposed into donkey-hide gelatin polypeptide under the action of the trichoderma CGMCC 3.3711 and the saccharomycete CMCC (B) 98007 during fermentation. As can be seen from fig. 1, the hydrolysis degree of donkey-hide gelatin is significantly reduced without using the trichoderma CGMCC 3.3711 bacterial liquid or the saccharomycete CMCC (B) 98007 bacterial liquid, which indicates that the significant synergistic effect between the trichoderma CGMCC 3.3711 and the saccharomycete CMCC (B) 98007 can promote the hydrolysis of donkey-hide gelatin, thereby obtaining more donkey-hide gelatin peptide.
Example 2
S1, pretreatment of donkey-hide gelatin: selecting high-quality donkey-hide gelatin as a raw material, ensuring the purity and quality of the donkey-hide gelatin, and crushing the donkey-hide gelatin into 400-500 mesh particles for later use;
s2, preparing Trichoderma CGMCC 3.3711 bacterial liquid: preparation of a concentration of 3X 10 9 Preparing cfu/ml trichoderma CGMCC 3.3711 bacterial liquid; preparation of saccharomycete CMCC (B) 98007 bacterial liquid: preparation of the concentration was 4.6X10 9 cfu/ml of saccharomycete CMCC (B) 98007 bacterial liquid for standby;
s3, pretreatment of rhizoma polygonati, chinese angelica, medlar, dendrobium candidum flowers, kudzuvine roots and ginger: pulverizing rhizoma polygonati, chinese angelica, medlar, dendrobium officinale flower, radix puerariae and ginger according to the same mass ratio, uniformly mixing, and mixing with water according to the mass ratio of 1:1.5, mixing, then adding the trichoderma liquid and the saccharomycete liquid prepared in the step S2, and carrying out jar fermentation, wherein the traditional Chinese medicine mixture is as follows: trichoderma liquid: the mass ratio of the saccharomycete liquid is 2:1:1, fermenting at 35 ℃ for 24 hours;
s4, filtering after fermentation, taking supernatant after filtering, pouring the supernatant into the donkey-hide gelatin melted in the step S1, uniformly stirring at the temperature of 35 ℃, and then keeping the temperature of 35 ℃ for fermentation for 12 hours to obtain donkey-hide gelatin polypeptide liquid containing traditional Chinese medicine fermentation liquid, wherein the fermentation liquid is prepared by the steps of: the mass ratio of the donkey-hide gelatin is 1:1, adding;
s5, after fermentation is completed, adding 3%H 2 O 2 Inactivating Trichoderma CGMCC 3.3711 and yeast CMCC (B) 98007, filtering the donkey-hide gelatin polypeptide liquid containing the Chinese medicinal fermentation liquid to remove solid particles and impurities, adding 2mM NaOH to precipitate donkey-hide gelatin polypeptide, and performingCentrifuging, collecting precipitate after centrifugation, and drying the collected precipitate at 30-40 ℃ to obtain the donkey-hide gelatin composite polypeptide for postpartum recovery.
Comparative example 3: the steps were the same as in example 2 except that the fermentation broth of the traditional Chinese medicine in step S4 was replaced with hot water for stirring.
Comparative example 4: the steps are the same as in example 2 except that the trichoderma CGMCC 3.3711 in step S2 is replaced with trichoderma CGMCC3.2942 (purchased from bio-technology limited of beijing hundred-ebo).
Comparative example 5: the steps were the same as in example 2 except that the yeast CMCC (B) 98007 in step S2 was replaced with yeast ATCC9763 (available from chu biotechnology (Jiangsu) limited).
FIG. 2 is a graph showing the comparison of the hydrolysis rate of colla Corii Asini in example 2 and comparative examples 3-5 according to the present invention. As can be seen from FIG. 2, in example 2, the decomposition rate can reach 67% by fermentation decomposition, and the hydrolysis rate is significantly reduced by hot water stirring, which shows the superiority of fungus decomposition. From the results of comparative example 4 and comparative example 3, it is understood that Trichoderma CGMCC 3.3711 and yeast CMCC (B) 98007 are specific to decomposing donkey-hide gelatin, and that the synergistic effect between the two bacteria is also specific.
Example 3
S1, pretreatment of donkey-hide gelatin: selecting high-quality donkey-hide gelatin as a raw material, ensuring the purity and quality of the donkey-hide gelatin, and crushing the donkey-hide gelatin into 400-500 mesh particles for later use;
s2, preparing Trichoderma CGMCC 3.3711 bacterial liquid: preparation of a concentration of 2.2X10 9 Preparing cfu/ml trichoderma CGMCC 3.3711 bacterial liquid; preparation of saccharomycete CMCC (B) 98007 bacterial liquid: preparation of the solution at a concentration of 3.9X10 9 cfu/ml of saccharomycete ATCC 12845 bacterial liquid for standby;
s3, pretreatment of rhizoma polygonati, chinese angelica, medlar, dendrobium candidum flowers, kudzuvine roots and ginger: pulverizing rhizoma polygonati, chinese angelica, medlar, dendrobium officinale flower, radix puerariae and ginger according to the same mass ratio, uniformly mixing, and mixing with water according to the mass ratio of 1:1.5, mixing, then adding the trichoderma liquid and the saccharomycete liquid prepared in the step S2, and carrying out jar fermentation, wherein the traditional Chinese medicine mixture is as follows: trichoderma liquid: the mass ratio of the saccharomycete liquid is 1:0.5:0.5, the fermentation temperature is 33 ℃, and the fermentation time is 18 hours;
s4, filtering after fermentation, taking supernatant, pouring the supernatant into the donkey-hide gelatin melted in the step S1, uniformly stirring at the temperature of 33 ℃, and then keeping the temperature of 32-35 ℃ for fermentation for 6-12 hours to obtain donkey-hide gelatin polypeptide liquid containing traditional Chinese medicine fermentation liquid, wherein the fermentation liquid is prepared by the steps of: the mass ratio of the donkey-hide gelatin is 1:1, adding;
s5, after fermentation is completed, adding 3%H 2 O 2 Inactivating Trichoderma CGMCC 3.3711 and saccharomycete CMCC (B) 98007, filtering the donkey-hide gelatin polypeptide liquid containing the Chinese medicine fermentation liquid to eliminate solid grains and impurity, adding 2mM NaOH to precipitate donkey-hide gelatin polypeptide, centrifuging to obtain precipitate, and drying the precipitate at 35 deg.c to obtain the composite donkey-hide gelatin polypeptide for post-partum recovery.
Comparative example 6: the steps are the same as example 3 except that no Chinese medicine is added for fermentation in step S3.
Comparative example 7: the steps are the same as those of example 3 except that kudzuvine root is not added in step S4.
Comparative example 8: the steps are the same as in example 3, except that no ginger is added in step S4.
Example 4
Animal experiment
Lactation conditioning experiment
Grouping, modeling and administration of animals
SPF-grade healthy KM breeds pregnant mice (supplied by Si Bei Fu (Beijing) biotechnology limited company, animal production license SCXK (Beijing) 2016-0002) 70, 130 days old, weight 92+ -11 g, and feeding in SPF-grade animal feeding room of the company, relatively constant temperature (25 ℃), constant humidity (75%), regular mouse pellet feed feeding, free ingestion and drinking water.
55 mice differing by not more than 24 h before and after the birth time were taken as experimental study subjects, and each litter was adjusted to 5 mice. The rats were randomly divided into a lack of milk model group, a normal group, a positive control estradiol group and an example group (example 3), and 11 groups of comparative examples (comparative example 1, comparative example 2, comparative example 4, comparative example 5, comparative example 6, comparative example 7 and comparative example 8) were used, each group being 5.
The day of delivery is 0 d, from 3 rd d after the delivery of the female mice, except for the normal group, the rest groups are filled with bromocriptine (1.6 mg/kg) daily to establish a model of postpartum hypogalactia of the mice, which is 10 d. At the time of molding, according to the human and animal dose change algorithm, example group (example 3), 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 respectively given example 3, comparative example 1, comparative example 2, comparative example 4, comparative example 5, comparative example 6, comparative example 7 and comparative example 8 prepared donkey-hide gelatin composite polypeptide 1.2g/100 g.d fed, and estradiol group was given 17 a-estradiol (1 g/kg), normal group and lack milk model group were given an equal amount of physiological saline. After gastric lavage 10 d (postpartum 13 th d), the female mice were sacrificed and blood and mammary tissue were taken for index detection. And (3) measuring the weight increment of the litter of the mice by referring to the method in the prior art, and detecting the lactation yield and the PRL.
(3) Index detection
Measurement of litter weight gain: the litter weight increase was 10 th d, and after the end, the last litter weight was weighed and the first litter weight was subtracted.
Lactation amount measurement: from the third day of dosing, fixed time day whole litter mice were weighed (W1), the mice were re-weighed after 4 h isolation from the master mice (W2), and then weighed a third time after 1 h of lactation with the master mice (W3). The hourly lactation amount of the female mice was calculated by taking the weight difference (W3-W2) of the whole litter mice before and after 1 hour of lactation, and adding the average basal metabolic amount ((W1-W2)/4) of the whole litter mice for 1 hour. I.e. milk yield per hour = w3-w2+ (W1-W2)/4.
The weight gain results of the litter are shown in Table 1:
TABLE 1
The lactation yield (mL) is shown in table 2:
TABLE 2
From the results of increasing litter weight and lactation yield of the mice in the milk deficiency model group in tables 1 and 2, respectively, it was found that the milk deficiency model was constructed successfully. From example 3, the results of comparative examples 1 and 2 show that the weight gain of litter and lactation yield of the mice are significantly reduced when fermentation is performed without using trichoderma CGMCC 3.3711 or yeast CMCC (B) 98007, indicating that the synergy between trichoderma CGMCC 3.3711 and yeast CMCC (B) 98007 can promote milk secretion. From example 3, comparative example 4 and comparative example 5, it is known that trichoderma CGMCC 3.3711 and yeast CMCC (B) 98007 are specific for promoting milk fermentation. According to example 3, comparative example 6, comparative example 7 and comparative example 8, the addition of the fermented product of the traditional Chinese medicine has unexpected effect of promoting the milk secretion, and the simultaneous addition of the kudzuvine root and the ginger in the 6 medicines can produce synergistic effect, wherein the kudzuvine root can promote the milk secretion, the ginger can drive the blood circulation, and the milk secretion is quickened.
The above examples merely illustrate specific embodiments of the invention, which are described in more detail and are not to be construed as limiting the scope of the invention. It should be noted that, it should be understood that those skilled in the art should not depart from the spirit and principle of the present invention, any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention are all equivalents to be included in the scope of the present invention.
Claims (9)
1. A composite microbial agent for fermenting donkey-hide gelatin is characterized in that: consists of Trichoderma CGMCC 3.3711 and saccharomycete CMCC (B) 98007.
2. The method for fermenting donkey-hide gelatin by using the composite microbial inoculum according to claim 1, which is characterized by comprising the following specific steps:
s1, pretreatment of donkey-hide gelatin: pulverizing colla Corii Asini;
s2, preparing Trichoderma CGMCC 3.3711 bacterial liquid: preparation of 1.5X10 concentration 9 ~3×10 9 Preparing cfu/ml trichoderma CGMCC 3.3711 bacterial liquid; preparation of saccharomycete CMCC (B) 98007 bacterial liquid: preparation of the concentration was 3.5X10 9 ~4.6×10 9 cfu/ml of saccharomycete CMCC (B) 98007 bacterial liquid for standby;
s3, pretreatment of rhizoma polygonati, chinese angelica, medlar, dendrobium candidum flowers, kudzuvine roots and ginger: pulverizing rhizoma polygonati, chinese angelica, medlar, dendrobium officinale flower, radix puerariae and ginger, uniformly mixing, and mixing with water according to a mass ratio of 1:1.5, adding the trichoderma liquid and the saccharomycete liquid prepared in the step S2, and carrying out jar fermentation at the temperature of 32-35 ℃ for 12-24 hours;
s4, filtering after fermentation, taking supernatant, adding the donkey-hide gelatin crushed in the step S1, stirring uniformly at the temperature of 32-35 ℃, and fermenting for 6-12 hours at the temperature of 32-35 ℃ to obtain donkey-hide gelatin polypeptide liquid containing traditional Chinese medicine fermentation liquid, wherein the fermentation liquid is prepared by the steps of: the mass ratio of the donkey-hide gelatin is 1:1, adding;
s5, after fermentation is completed, adding 3%H 2 O 2 Inactivating Trichoderma CGMCC 3.3711 and saccharomycete CMCC (B) 98007, filtering donkey-hide gelatin polypeptide liquid containing Chinese medicine fermentation liquid to eliminate solid grains and impurity, adding proper amount of 2mM NaOH to precipitate donkey-hide gelatin polypeptide, centrifuging to obtain precipitate, and drying the precipitate at 30-40 deg.c to obtain the composite donkey-hide gelatin polypeptide for post partum recovery.
3. The method for fermenting donkey-hide gelatin by using the composite microbial agent according to claim 2, wherein: the concentration of the Trichoderma CGMCC 3.3711 bacterial liquid used in the step S2 is 2.2X10 9 cfu/ml。
4. The method for fermenting donkey-hide gelatin by using the composite microbial agent according to claim 3, wherein: the concentration of the saccharomycete CMCC (B) 98007 bacterial liquid used in the step S2 is 3.9X10 9 cfu/ml。
5. The method for fermenting donkey-hide gelatin by using the composite microbial agent according to claim 4, wherein the method comprises the following steps: and step S3, grinding the rhizoma polygonati, the Chinese angelica, the medlar, the dendrobium candidum flowers, the kudzuvine roots and the ginger according to the same mass ratio, and uniformly mixing.
6. The method for fermenting donkey-hide gelatin by using the composite microbial agent according to claim 4, wherein the method comprises the following steps: in the step S3, the Chinese medicine mixture: trichoderma liquid: the mass ratio of the saccharomycete liquid is 1:0.5:0.5, mixing evenly.
7. The method for fermenting donkey-hide gelatin by using the composite microbial agent according to claim 6, wherein: and (3) directly mixing the supernatant obtained in the step (S4) with the melted donkey-hide gelatin obtained in the step (S1) for fermentation without inactivating.
8. The donkey-hide gelatin composite polypeptide prepared by the method for fermenting donkey-hide gelatin by using the composite microbial inoculum according to any one of claims 3-7.
9. A preparation method of donkey-hide gelatin paste is characterized in that: the use of a donkey-hide gelatin composite polypeptide as claimed in claim 8 for preparing a donkey-hide gelatin composite polypeptide paste;
the preparation method comprises the following steps:
heating the high-quality donkey-hide gelatin block at 80-90 ℃ and then melting, keeping the temperature at 60-70 ℃ after melting, and keeping a melting state, wherein the mass ratio of the donkey-hide gelatin block to the high-quality donkey-hide gelatin block is 1:0.5 adding the donkey-hide gelatin composite polypeptide prepared by the invention, and then mixing the donkey-hide gelatin composite polypeptide with high-quality donkey-hide gelatin according to a mass ratio of 1:0.5:0.2:0.2: and 0.1, respectively adding black sesame, medlar, red date and walnut, uniformly mixing and stirring, spreading and cooling, and cutting into small blocks with the length of 6-8cm, the width of 3-4cm and the thickness of 1-1.5cm to obtain the donkey-hide gelatin polypeptide paste.
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