Fermented soybean peptide, fermentation process of high-quality fermented soybean and fermented soybean primary fermentation equipment
By optimizing the fermentation process and equipment of light fermented black beans, high-quality peptides were extracted and isolated, and the existing problems of unstable quality and limited yield of light fermented black beans were solved, which significantly improved the medicinal efficacy of light fermented black beans, especially in anti-pulmonary fibrosis.
Patent Information
- Application Number
- CN202411214142.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-08-31
- Publication Date
- 2025-04-29
AI Technical Summary
The existing fermentation process and equipment of light fermented black beans have problems of unstable quality and limited yield, and the research focus is mainly on small-molecular compounds such as flavonoids and saponins, and the protein and peptide components in light fermented black beans are not fully utilized.
The optimized fermentation process of light fermented black beans and a primary fermentation equipment was adopted to obtain the fermented black beans and proteins through alkali acid extraction, and the protease enzymatic separation was carried out to obtain 1000-3000 Da or 3000-5000 Da polypeptides, and a single fermentation equipment of light fermented black beans was designed to control the fermentation temperature and humidity.
The stable production of high-quality light fermented black beans has been achieved, which significantly improves the medicinal efficacy of light fermented black beans, especially in anti-pulmonary fibrosis.
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Figure CN119899251A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the field of pharmaceutical chemistry and pharmaceutical machinery, and relates to the discovery, processing technology and processing equipment of active ingredients of Chinese herbal medicine slices, and specifically to a light fermented black bean peptide, a fermentation technology of high-quality light fermented black bean and light fermented black bean primary fermentation equipment. Background Art
[0002] Light black bean curd is a fermented product of the mature seeds (black beans) of the leguminous plant soybean Glycine max (L.) Merr. It is bitter, pungent, and cool in nature. It belongs to the lung and stomach meridians. It has the effects of dispelling exterior pathogens, eliminating restlessness, and dispelling stagnation and heat. It can be used for colds, cold and heat headaches, irritability, chest tightness, and insomnia. Light black bean curd contains flavonoids, saponins, proteins, polysaccharides, amino acids, fibrinolytic enzymes and other ingredients, among which flavonoids are considered to be the main active ingredients. Light black bean curd has pharmacological activities such as lowering blood sugar, regulating intestinal flora, antibacterial, anti-osteoporosis and anti-cancer (Reference: Cao Dongying et al., Analysis of isoflavone contents in 4 commercially available black beans and finished light black bean curd, Pharmaceutical Research, 2020). Cold medicines such as Antelope Cold Soft Capsules, Vitamin C Yinqiao Tablets, and Yinqiao Detoxification Capsules all contain light black bean curd.
[0003] There are four types of black beans commonly used in commercially available light black beans, including small black beans with yellow heart, black kidney beans, green black beans, and large black beans with yellow heart, all of which are authentic black beans. Among them, small black beans with yellow heart are used in the traditional customs of Anguo, and small black beans with yellow heart are also the main raw materials in the traditional preparation method of light black beans in Fuzhou. During the fermentation process of light black beans, when the surface mycelium of several types of black beans is washed off, the texture of small black beans with yellow heart is the hardest, not easy to peel, and more conducive to maintaining the appearance of the finished product. Therefore, small black beans with yellow heart are used as the better raw material for the fermentation of light black beans (Reference: Cao Dongying et al., Analysis of isoflavones content in 4 commercially available black beans and finished light black beans, Pharmaceutical Research, 2020).
[0004] Traditional light black beans are mainly fermented by natural fermentation, with multiple strains participating in the fermentation, and the fermentation bacteria mainly come from the air. After modern scholars studied the flora of naturally fermented light black beans, they found that the fermentation bacteria involved include Aspergillus, Rhizopus, Mucor, Saccharomyces, Penicillium, Micrococcus, and Cocci. Among them, bacteria (mainly Bacillus subtilis) and molds (Rhizopus, Aspergillus, and Mucor) are common fermentation strains and are the dominant fermentation bacteria; while the mechanism of action of cocci, micrococci, etc. on the fermentation of black beans is still unclear, and they should not be the dominant flora (Reference: Lin Wangmin et al., Overview of the Fermentation Process Evolution and Process Control of Light Black Beans, Chinese Journal of Experimental Traditional Chinese Medicine, 2021).
[0005] In the production process of light fermented black beans, fermentation technology and process control are the key to determining the quality of the finished medicinal materials. The 2020 edition of the "Chinese Pharmacopoeia" stipulates the preparation method: take 70-100g of mulberry leaves and Artemisia annua, add water to boil, filter, mix the decoction into 1000g of clean soybeans, wait until it is completely absorbed, steam it thoroughly, take it out, dry it slightly, and then put it in a container, cover it with the decocted mulberry leaves and Artemisia annua residue and ferment it until the yellow coat is on the top, take it out, remove the residue, wash it, put it in a container and then simmer it for 15-20 days, until it is fully fermented and the aroma overflows, take it out, steam it slightly, and dry it. It can be seen that the 2020 edition of the "Chinese Pharmacopoeia" uses Artemisia annua and mulberry leaves as auxiliary materials and uses the secondary fermentation method to produce light fermented black beans, but there is no clear regulation on the temperature and humidity of the pre-fermentation and post-fermentation (re-simmering). Modern research has few differences in the pre-fermentation process. The fermentation temperature is mostly 25-32℃, the relative humidity is about 80%, and the fermentation time is 4-7d. However, the post-fermentation temperature selected by different researchers and manufacturers varies greatly, ranging from 30 to 60°C, and the post-fermentation time is 12 to 20 days (reference: Zhong Rongrong et al., Study on Influencing Factors in the Fermentation Process of Light Douchi, Chinese Patent Medicine, 2024). This leads to the uneven quality of light Douchi on the market, and the efficacy cannot be guaranteed.
[0006] The primary fermentation (pre-fermentation) is one of the important process steps of fermentation of light fermented black beans. At present, the existing technology mainly uses handmade products such as bamboo plaques and bamboo baskets for primary fermentation, which are all manually operated. It is impossible to control the growth state of Aspergillus to meet the requirements of yellow coat in batches, and the production capacity is small, which limits the quality and output of light fermented black beans.
[0007] In addition, according to records, the protein content in black beans is as high as 36% to 40%, and the light black bean sauce prepared from black beans also contains a high content of protein (reference: Zhao Chaofan et al., Effects of different fermentation methods on the quality and flavor of light black bean sauce, Food Industry Science and Technology, 2022). Compared with black beans, light black bean sauce undergoes steaming and fermentation, and the protein is modified, which will produce active and diverse proteins and peptides. Unfortunately, the current research focus of both academia and industry is still on small molecule compounds such as flavonoids and saponins in light black bean sauce.
[0008] In order to solve the above problems, the present invention is proposed. Summary of the invention
[0009] The purpose of the present invention is to overcome the deficiencies of the prior art and provide a light fermented soybean peptide, a fermentation process of high-quality light fermented soybean and a light fermented soybean primary fermentation device.
[0010] The above-mentioned purpose of the present invention is achieved through the following technical solutions:
[0011] A polypeptide obtained from high-quality light fermented black beans, the preparation method comprising the following steps:
[0012] Step 1: Extract light fermented black bean protein by alkali extraction and acid precipitation
[0013] Take crushed and defatted light fermented black beans, add appropriate amount of purified water, adjust pH=10.8, extract in a water bath for a certain period of time, cool, centrifuge, take the supernatant, adjust pH=4.6, centrifuge, collect the precipitate and disperse it in appropriate amount of purified water, adjust pH=7.0, dialyze for desalination, freeze-dry, and obtain light fermented black bean protein;
[0014] Step 2: Protease digestion
[0015] Add appropriate amount of purified water to the light fermented black bean protein, stir to dissolve, add appropriate amount of alkaline protease for enzymolysis, enzymolysis temperature = 50°C, enzymolysis pH = 10.0, enzymolysis time = 2.5h, inactivate the enzyme in a boiling water bath, centrifuge, take the supernatant, dialyze for desalination, and freeze-dry to obtain light fermented black bean protein hydrolysate;
[0016] Step 3: Ultrafiltration to separate peptides of different molecular weights
[0017] Add appropriate amount of purified water to the light fermented black bean protein hydrolysate, stir to dissolve, and ultrafilter with ultrafiltration membranes of specifications of 5000Da, 3000Da, and 1000Da in sequence to collect polypeptide solutions of 1000-3000Da and 3000-5000Da, respectively, and freeze-dry.
[0018] Preferably, sodium hydroxide solution and hydrochloric acid solution are used to adjust the pH.
[0019] Preferably, the centrifugation parameters are all 4000r / min for 20min.
[0020] Preferably, 1 g of light fermented black bean protein is added with 4000 U of alkaline protease.
[0021] Preferably, the preparation method of the high-quality light fermented black beans comprises: taking 70-100 parts by weight of mulberry leaves and Artemisia annua, boiling with water, filtering, mixing the decoction into 1000 parts by weight of black beans, the weight ratio of the decoction to the black beans is 1-1.2:1, mixing the decoction and the black beans evenly, placing them in a steaming container after they are completely absorbed, steaming them thoroughly at 103-108°C, taking them out, cooling them slightly, and then placing them in a fermentation container, maintaining the fermentation temperature at 26-29°C and the humidity at 80-85%, fermenting for 2-3 days, taking them out when the yellow skin appears, washing them, placing them in a fermentation container at 50-55°C for another 15-20 days, taking them out when they are fully fermented and the aroma overflows, steaming them briefly at 100-105°C, and drying them.
[0022] More preferably, the steaming is performed at 103-108° C. for 40 min.
[0023] More preferably, the brief steaming is steaming at 100-105° C. for 25 min.
[0024] A light fermented black bean curd primary fermentation device, used as the above-mentioned fermentation device with a fermentation temperature of 26 to 29° C., comprises a feeding mechanism and a fermentation mechanism; wherein:
[0025] The feeding mechanism includes a feeding machine for evenly feeding the steamed black beans to the fermentation mechanism and a distributing machine for evenly distributing the steamed black beans to the feeding machine;
[0026] The fermentation mechanism includes a box body, a humidifier and a dehumidification fan arranged in the box body for controlling the fermentation humidity, a heat pump main unit arranged in the box body for controlling the fermentation temperature, and a fermentation plate connection arranged in the box body and rotating at a constant speed to receive black beans from a feeder until the black beans are fully covered, and one end of the fermentation plate connection is located below the feeder.
[0027] Preferably, the fermentation mechanism is further provided with a camera and a lighting device. The lighting device can provide lighting for the box, and the camera can collect images of whether the fermentation plate is fully covered with black beans, so as to determine whether the fermentation plate is rotated or stopped.
[0028] The polypeptide is used for preparing medicine for treating pulmonary fibrosis.
[0029] Beneficial effects:
[0030] 1. The present invention optimizes a preparation method of high-quality light fermented black beans based on the preparation method specified in the 2020 edition of the Chinese Pharmacopoeia. The fermentation (primary fermentation, secondary fermentation) process parameters in the preparation method are specific and clear, and high-quality light fermented black beans can be stably obtained; the present invention also provides a light fermented black bean primary fermentation equipment for the primary fermentation process in the preparation method, and the primary fermentation equipment can effectively control the temperature and humidity of the primary fermentation and can be used for mass production, overcoming the defects of the prior art;
[0031] 2. Light fermented black beans are a Chinese medicine with the functions of dispelling exterior pathogens, eliminating restlessness, and dispelling stagnation and heat, and can be used for colds, headaches due to cold and heat, chest tightness due to irritability, insomnia due to deficiency, etc. The flavonoid components contained in them are considered to be the main active ingredients. Colds are often accompanied by respiratory tract infections, and severe lower respiratory tract infections can induce pulmonary fibrosis. The present invention obtains a polypeptide with a weight average molecular weight of 1000-3000Da or 3000-5000Da from the above-mentioned high-quality light fermented black beans, and the polypeptide has a significant anti-pulmonary fibrosis effect. BRIEF DESCRIPTION OF THE DRAWINGS
[0032] Figure 1 The present invention provides a schematic structural diagram of the light black bean primary fermentation equipment; wherein, 1, a box body; 2, a heat pump host; 3, a humidifier; 4, a dehumidification fan; 5, a fermentation plate; 6, a feeder; 7, a material distributor. DETAILED DESCRIPTION
[0033] The essential contents of the present invention are described in detail below in conjunction with the embodiments, but the protection scope of the present invention is not limited thereto.
[0034] Embodiment 1: the preparation technology of high-quality light fermented black beans
[0035] The 2020 edition of the "Chinese Pharmacopoeia" stipulates the preparation method of light fermented black beans: take 70-100g each of mulberry leaves and Artemisia annua, add water and boil, filter, mix the decoction into 1000g of clean soybeans, wait until it is completely absorbed, steam it thoroughly, take it out, let it dry slightly, put it in a container, cover it with the boiled mulberry leaves and Artemisia annua residue and ferment it until it turns yellow, take it out, remove the residue, wash it, put it in a container and let it ferment for another 15-20 days, until it is fully fermented and the aroma overflows, take it out, steam it slightly, and dry it.
[0036] The preparation method of the present invention is optimized on the basis of the preparation method specified in the 2020 edition of the "Chinese Pharmacopoeia", and is specifically as follows: take 70-100g of mulberry leaves and Artemisia annua, add water to boil, filter, mix the decoction into 1000g of clean black beans, the ratio of the decoction to the black beans is 1-1.2:1 (weight ratio), the decoction and the black beans are mixed evenly, and after they are completely absorbed, they are placed in a steaming container, steamed thoroughly (steamed at 103-108°C for 40min), taken out, slightly cooled, and then placed in a fermentation container (primary fermentation, also known as pre-fermentation), maintaining the fermentation temperature at 26-29°C and the humidity at 80-85%, fermenting for 2-3 days, taking out when the yellow coat appears, washing, placing in a fermentation container and then stuffing for 15-20 days (50-55°C, secondary fermentation, also known as post-fermentation), until it is fully fermented and the aroma overflows, taking out, slightly steaming (steaming at 100-105°C for 25min), and drying to obtain.
[0037] The black beans used in the above-mentioned preparation method are small black beans with yellow core, and the reasons for preference are shown in the background technology.
[0038] The weight and volume in the above preparation method are for simplicity and convenience as in the Chinese Pharmacopoeia. Those skilled in the art should understand that in actual operation, they are not limited to specific weight and volume values, but the proportion of different materials is determined based on the above specific weight and volume values.
[0039] When each parameter in the above preparation method is expressed as a range, it means that high-quality light black beans can be obtained when the parameter is within the range.
[0040] Embodiment 2: Primary fermentation equipment of high-quality light fermented black beans
[0041] As described in the background technology, in the prior art, primary fermentation mostly uses handmade products such as bamboo plaques and bamboo baskets, which are all manually operated. It is impossible to control the growth state of Aspergillus to meet the requirement of yellow coat in batches, and the production capacity is small, which limits the quality and yield of light fermented black beans. This embodiment provides a primary fermentation device for primary fermentation in Example 1.
[0042] like Figure 1The light black bean fermentation equipment shown is used as the fermentation equipment with a fermentation temperature of 26 to 29° C., including a feeding mechanism and a fermentation mechanism; wherein:
[0043] The feeding mechanism includes a feeding machine 6 for evenly feeding the steamed black beans to the fermentation mechanism and a distribution machine 7 for evenly distributing the steamed black beans to the feeding machine 6;
[0044] The fermentation mechanism includes a box body 1, a humidifier 3 and a dehumidification fan 4 arranged in the box body 1 for controlling the fermentation humidity, a heat pump main unit 2 arranged in the box body 1 for controlling the fermentation temperature, and a fermentation plate 5 arranged in the box body 1 for rotating at a constant speed to receive black beans from a feeder 6 until they are fully covered, and one end of the fermentation plate 5 is located below the feeder 6. A camera device and a lighting device are also arranged in the fermentation mechanism. The lighting device can provide lighting in the box body, and the camera device can capture images of whether the fermentation plate is fully covered with black beans, so as to determine the rotation and stop of the fermentation plate.
[0045] Working principle: The steamed black beans are evenly spread on the feeder through a spreading machine, and the fermentation plate receives the black beans from the feeder evenly while rotating at a uniform speed. The feeding is stopped when the black beans are evenly spread on the fermentation plate. The temperature in the box is controlled within the temperature range specified by the process through the heat pump host, and the humidity in the box is controlled within the humidity range specified by the process through the humidifier and the dehumidification fan, so that the black beans are fermented under the temperature and humidity conditions specified by the process. When the black beans are fermented to the yellow coat, they are taken out. The lighting equipment is used to provide lighting in the box, and the camera equipment is used to collect images of whether the fermentation plate is covered with black beans and transmit them to the control mechanism to control the feeding to stop. The one-time fermentation equipment can effectively control the temperature and humidity of the one-time fermentation and can be used for mass production, overcoming the defects of the prior art.
[0046] Example 3: Discovery of active peptides in high-quality light fermented black beans
[0047] 1. Materials
[0048] High-quality light fermented black beans were used as raw materials, and the raw materials were prepared according to the preparation process parameters of Example 1, and the parameters involved in the range selected the middle value (different parameters in the range were not obviously different). The raw materials were crushed and then subjected to heat reflux defatting treatment with food-grade 95% ethanol (volume percentage concentration) (1kg:10L, 2.5h each time, repeated 2 times) and then used for the preparation of active peptides.
[0049] Alkaline protease (200,000 U / g), Beijing Solebow Technology Co., Ltd.
[0050] Dialysis bag (molecular weight cut-off > 500Da), Shanghai Jizhi Biochemical Technology Co., Ltd.
[0051] Human embryonic lung fibroblast MRC-5, Shenzhen Haodi Huatuo Biotechnology Co., Ltd.
[0052] 2. Methods
[0053] 1. Preparation of active peptides
[0054] (1) Extraction of light fermented black bean protein by alkali extraction and acid precipitation
[0055] Take crushed and defatted light black beans, add purified water (Wahaha, 1 g: 20 mL), adjust pH to 10.8 with sodium hydroxide solution (1 mol / L), extract for 3.5 h in a 65°C water bath, cool, centrifuge at 4000 r / min for 20 min, take the supernatant, adjust pH to 4.6 with hydrochloric acid solution (1 mol / L), centrifuge at 4000 r / min for 20 min, collect the precipitate and disperse it in an appropriate amount of purified water, adjust pH to 7.0 with sodium hydroxide solution (1 mol / L), dialyze for desalination, and freeze-dry to obtain light black bean protein.
[0056] (2) Protease hydrolysis
[0057] Add purified water (Wahaha, 1 g: 20 mL) to the light fermented black bean protein, stir to dissolve, add alkaline protease (1 g of light fermented black bean protein plus 4000 U of alkaline protease) for enzymatic hydrolysis (temperature = 50°C, pH = 10.0, time = 2.5 h), inactivate the enzyme in a boiling water bath for 10 min, centrifuge at 4000 r / min for 20 min, take the supernatant, dialyze for desalination, and freeze-dry to obtain the light fermented black bean protein hydrolysate.
[0058] (3) Ultrafiltration separation of peptides of different molecular weights
[0059] Purified water (Wahaha, 1 g: 30 mL) was added to the light fermented black bean protein hydrolysate, and the solution was stirred to dissolve. The solution was then ultrafiltered with ultrafiltration membranes of 5000 Da, 3000 Da, and 1000 Da in sequence to obtain polypeptide solutions of four weight-average molecular weight (Mw) components (<1000 Da, 1000-3000 Da, 3000-5000 Da, and >5000 Da), which were then freeze-dried.
[0060] 2. Anti-pulmonary fibrosis activity of active peptides
[0061] (1) Cell culture
[0062] Human embryonic lung fibroblasts MRC-5 were revived and cultured in MEM containing 10% fetal bovine serum, 2 mmol / L L-glutamine, 100 U / mL penicillin and 100 mg / L streptomycin at 37°C, saturated humidity, and 5% CO. 2 The cells were cultured and passaged under the appropriate conditions, and the MRC-5 cells with good growth status were selected for subsequent experiments.
[0063] (2) Experimental groups
[0064] MRC-5 cells were divided into blank control group, model group (20 μg / LTGF-β1), and peptide group (20 μg / mL, 40 μg / mL peptides with different weight-average molecular weights + 20 μg / LTGF-β1).
[0065] (3) CCK-8 detection of cell activity
[0066] MRC-5 cells in good growth state were inoculated into 96-well plates at 5000 cells / 100 μL culture medium per well and cultured for 24 h. The cells were treated according to the above grouping. 100 μL of fresh complete culture medium was added to the blank control group, 100 μL of complete culture medium containing TGF-β1 was added to the model group so that the final concentration of TGF-β1 was 20 μg / L, and 100 μL of complete culture medium containing TGF-β1 and polypeptides of different weight-average molecular weights was added to the polypeptide group so that the final concentration of TGF-β1 was 20 μg / L and the final concentration of polypeptide was 20 μg / mL or 40 μg / mL. The cells were cultured at 37°C, saturated humidity, and 5% CO. 2 After 48 h of continuous culture, 10 μL of CCK-8 solution was added to each well and incubated for 4 h. The absorbance (OD) value was read at 450 nm in a microplate reader. The cell activity of the blank control group was taken as 100% to calculate the cell activity of the model group and the polypeptide group.
[0067] (4) Data processing
[0068] The data were expressed as mean ± SD, and the t test was used for comparison among the groups. P < 0.05 indicated a significant difference.
[0069] 3. Results
[0070] Abnormal proliferation of lung fibroblasts is an important feature of pulmonary fibrosis, and inhibiting abnormal proliferation of lung fibroblasts is an important means of treating pulmonary fibrosis. The cell activities of each group are shown in Table 1. Compared with the blank control group, the cell activity of human embryonic lung fibroblasts MRC-5 in the model group was significantly increased (P<0.05), indicating that TGF-β1 induced successful pulmonary fibrosis modeling; compared with the model group, the cell activity of 1000-3000Da and 3000-5000Da polypeptide groups was significantly decreased (P<0.05), indicating that 1000-3000Da and 3000-5000Da polypeptides can inhibit abnormal proliferation of MRC-5 cells in a dose-dependent manner. The other two weight-average molecular weight polypeptides had no obvious inhibitory effect on the proliferation of MRC-5 cells.
[0071] Table 1 MRC-5 cell activity
[0072]
[0073] * represents P < 0.05 compared with the blank control group, # represents P < 0.05 compared with the model group.
[0074] Light Douchi is a traditional Chinese medicine that has the functions of dispelling exterior pathogens, eliminating restlessness, and dispelling stagnation and heat. It can be used for colds, headaches due to cold and heat, chest tightness due to restlessness, insomnia due to deficiency, etc. The flavonoids it contains are considered to be the main active ingredients. Colds are often accompanied by respiratory tract infections, and severe lower respiratory tract infections can induce pulmonary fibrosis. The above experiments have proved that the polypeptides with a weight average molecular weight of 1000-3000Da and 3000-5000Da obtained from light Douchi have obvious anti-pulmonary fibrosis effects.
[0075] The purpose of the above-mentioned embodiments is to specifically introduce the essential content of the present invention, but those skilled in the art should know that the protection scope of the present invention should not be limited to the specific embodiments.
Claims
1. A polypeptide obtained from high-quality light fermented black beans, characterized in that: The preparation method comprises the following steps: Step 1: Extract light fermented black bean protein by alkali extraction and acid precipitation Take crushed and defatted light fermented black beans, add appropriate amount of purified water, adjust pH=10.8, extract in a water bath for a certain period of time, cool, centrifuge, take the supernatant, adjust pH=4.6, centrifuge, collect the precipitate and disperse it in appropriate amount of purified water, adjust pH=7.0, dialyze for desalination, freeze-dry, and obtain light fermented black bean protein; Step 2: Protease digestion Add appropriate amount of purified water to the light fermented black bean protein, stir to dissolve, add appropriate amount of alkaline protease for enzymolysis, enzymolysis temperature = 50°C, enzymolysis pH = 10.0, enzymolysis time = 2.5h, inactivate the enzyme in a boiling water bath, centrifuge, take the supernatant, dialyze for desalination, and freeze-dry to obtain light fermented black bean protein hydrolysate; Step 3: Ultrafiltration to separate peptides of different molecular weights Add appropriate amount of purified water to the light fermented black bean protein hydrolysate, stir to dissolve, and ultrafilter with ultrafiltration membranes of specifications of 5000Da, 3000Da, and 1000Da in sequence to collect polypeptide solutions of 1000-3000Da and 3000-5000Da, respectively, and freeze-dry.
2. The polypeptide according to claim 1, characterized in that: Sodium hydroxide solution and hydrochloric acid solution were used to adjust pH.
3. The polypeptide according to claim 1, characterized in that: The centrifugation parameters were all 4000r / min for 20min.
4. The polypeptide according to claim 1, characterized in that: 1g of light fermented black bean protein plus 4000U of alkaline protease.
5. The polypeptide according to claim 1, characterized in that The preparation method of high-quality light fermented black beans comprises: taking 70-100 parts by weight of mulberry leaves and 70-100 parts by weight of wormwood, boiling with water, filtering, mixing the decoction into 1000 parts by weight of black beans, wherein the weight ratio of the decoction to the black beans is 1-1.2:1, mixing the decoction and the black beans evenly, placing them in a steaming container after they are completely absorbed, steaming them thoroughly at 103-108° C., taking them out, cooling them slightly, placing them in a fermentation container again, maintaining the fermentation temperature at 26-29° C., the humidity at 80-85%, fermenting them for 2-3 days, taking them out when they turn yellow, washing them, placing them in a fermentation container at 50-55° C. for another 15-20 days, taking them out when they are fully fermented and the aroma overflows, steaming them slightly at 100-105° C., and drying them to obtain the product.
6. The polypeptide according to claim 5, characterized in that: The steaming is performed at 103-108° C. for 40 minutes.
7. The polypeptide according to claim 5, characterized in that: The brief steaming is steaming at 100-105° C. for 25 minutes.
8. A primary fermentation device for light fermented black beans, used as the fermentation device with a fermentation temperature of 26 to 29° C. in claims 1 to 7, characterized in that: It includes a feeding mechanism and a fermentation mechanism; wherein: The feeding mechanism includes a feeding machine for evenly feeding the steamed black beans to the fermentation mechanism and a distributing machine for evenly distributing the steamed black beans to the feeding machine; The fermentation mechanism includes a box body, a humidifier and a dehumidification fan arranged in the box body for controlling the fermentation humidity, a heat pump main unit arranged in the box body for controlling the fermentation temperature, and a fermentation plate connection arranged in the box body and rotating at a constant speed to receive black beans from a feeder until the black beans are fully covered, and one end of the fermentation plate connection is located below the feeder.
9. The light fermented black bean curd primary fermentation equipment according to claim 8, characterized in that: The fermentation mechanism is also provided with a camera and a lighting device.
10. Use of the polypeptide according to any one of claims 1 to 7 for preparing a drug for treating pulmonary fibrosis.