Preparation method and application of compound bacteria for simultaneous fermentation of Rehmannia glutinosa with laxative function

Abstract

This invention relates to the field of microbial fermentation technology, specifically a method for preparing Rehmannia glutinosa through simultaneous fermentation with compound bacteria and a laxative effect. This method involves extracting four types of probiotics that meet national safety standards: Lactobacillus acidophilus, Bifidobacterium longum, Lactobacillus casei, and Saccharomyces cerevisiae, and then formulating a high-concentration compound bacterial agent according to the live bacteria ratio. The initial fermentation material is adjusted to a material-to-water ratio of 1:5 and a suitable pH is maintained. After autoclaving, the materials are inoculated with bacteria at a weight-to-volume ratio for biological culture. The entire system is placed in a 37°C environment for continuous constant-temperature static co-fermentation. Through specific enzymatic hydrolysis and metabolic pathways of multidimensional bacterial communities, the absolute content of the core active ingredient catalpol and soluble dietary fiber in the final product is significantly increased. This bio-fermentation process effectively eliminates the potential risk of spleen and stomach irritation and damage from traditional Chinese medicinal materials. The resulting dry powder raw material can be widely and safely used in the preparation of microecological functional foods or pharmaceutical preparations targeting constipation symptoms caused by intestinal dryness and fluid deficiency.

Description

Technical Field

[0001] This invention relates to the field of microbial fermentation technology, specifically to a method and application of simultaneous fermentation of Rehmannia glutinosa by compound bacteria with laxative function. Background Technology

[0002] Rehmannia glutinosa, as a traditional Chinese medicinal herb and a food-medicine homology substance, has a long history of application in nourishing yin, clearing heat, promoting body fluids, and moistening dryness. Modern microecological theory points out that combining Chinese medicinal herbs with probiotics for biotransformation can effectively degrade large, poorly absorbed molecules in the herbs while retaining and enhancing their core active ingredients. This provides a core research and development direction for developing modern, novel microecological Chinese medicine products with laxative functions.

[0003] There are currently published exploratory studies on the fermentation of Rehmannia glutinosa using a single strain. A search reveals Chinese Patent Publication No. CN120713964A, which discloses a fermented extract of Rehmannia glutinosa and its preparation method. This technique relies solely on a single strain of *Aspergillus cristatus* to ferment Rehmannia glutinosa, and its core objective remains at the level of increasing polysaccharide content in a single dimension. Existing conventional fermentation extraction techniques typically remain at a superficial transformation stage, lacking the simulation and adaptation to the complex intestinal microecological environment.

[0004] The aforementioned existing technologies have extremely obvious limitations and technical defects. A single strain of bacteria cannot completely degrade the dense cellulose and pectin and other large molecular wall substances inside Rehmannia glutinosa, resulting in extremely insufficient release of effective ingredients; the function of fermentation products is too singular, and the increase in characteristic active ingredients such as catalpol, which are highly related to bowel movement, is weak, lacking targeted efficacy optimization design for constipation pathology; the existing process completely ignores the synergistic metabolic mechanism between multiple strains and fails to systematically enhance the laxative efficacy of fermentation products through the optimal configuration of live bacteria ratio. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention provides a method and application for preparing and processing Rehmannia glutinosa through simultaneous fermentation of compound bacteria with laxative function. This method solves the problems of incomplete fermentation of single strains, insufficient targeted efficacy, and potential stomach and spleen discomfort caused by traditional processing methods in existing technologies.

[0006] To achieve the above objectives, the present invention provides the following technical solution: a method for preparing Rehmannia glutinosa through simultaneous fermentation by compound bacteria with laxative function, comprising the following steps: S1. Extract Lactobacillus acidophilus CICC 6074, Bifidobacterium longum CICC 6188, Lactobacillus casei CICC 6096 and Saccharomyces cerevisiae CICC 31512 at a predetermined concentration, and aseptically mix the bacterial suspensions of the above four microorganisms in a ratio of 3:2:2:1 to construct a compound microbial agent. S2. Take Rehmannia glutinosa powder as the substrate raw material, add sterile purified water according to the predetermined feed water ratio, adjust the pH of the mixed system and then perform high-temperature steam sterilization. After the system is cooled to the appropriate inoculation temperature, the fermentation substrate is obtained. S3. The aforementioned compound microbial agent is added to the fermentation substrate according to the volume-mass ratio. After being thoroughly mixed, the mixture is subjected to static synchronous compound fermentation under constant temperature conditions. S4. After the fermentation cycle is completed, the microbial metabolic process is completely terminated by high-temperature insulation. The resulting material is then transferred to a vacuum environment for low-temperature drying until it reaches a constant weight. Finally, it is pulverized and sieved to obtain synchronous compound fermented Rehmannia glutinosa powder.

[0007] Preferably, the total viable count in the compound microbial agent reaches 5 × 10⁻⁶. 9 For strains at CFU / mL or higher, each individual bacterial species constituting the microbial agent must undergo activation and expansion culture procedures beforehand. Lactic acid bacteria are activated on MRS medium in an anaerobic environment at 37°C for 18 hours and expanded culture for 24 hours, respectively. Saccharomyces cerevisiae is activated on YPD medium at 30°C with shaking for 24 hours and expanded culture with aeration for 36 hours. The concentration of each individual bacterial suspension is uniformly adjusted to 1×10⁻⁶ before mixing. 9 CFU / mL.

[0008] Preferably, the processing procedure for the Rehmannia glutinosa powder is as follows: select Rehmannia glutinosa from Jiaozuo, Henan Province, that is free from mold and insect infestation, rinse it three times with flowing purified water to remove impurities, then cut it into thin slices with a thickness of 3-5 mm, place the slices in a 50°C forced-air drying oven to dry until the moisture content is less than 10% as detected by a moisture analyzer, and use a mechanical pulverizing device to pulverize the dried material and make it completely pass through a 60-mesh standard sieve.

[0009] Preferably, during the preparation of the fermentation substrate, the ratio of Rehmannia glutinosa powder to sterile purified water is set to 1:5 (w / v). A 1 mol / L HCl or NaOH solution is added dropwise to adjust the pH of the substrate system to within the range of 6.5±0.1. The sterilization program uses 121℃ and 0.1MPa high-pressure steam for 15 minutes. After the sterilization is completed, the temperature drops rapidly to 37℃ at a cooling water circulation system at a cooling rate of 5℃ / min.

[0010] Preferably, the inoculum amount of the compound microbial agent is 5% (v / w) of the total weight of the fermentation substrate. After inoculation, a sterile agitator should be started and continuously stirred at 80 rpm for 5 minutes to ensure uniform dispersion. The entire fermentation process is maintained at a constant temperature of 37°C for up to 72 hours. During this period, samples should be taken every 12 hours to confirm that the pH value of the system is maintained at 4.1±0.2 and the overall viable cell count is maintained at ≥9×10⁻⁶.9 Above the standard of CFU / mL.

[0011] Preferably, the specific process conditions for the fermentation termination and post-processing stages are as follows: the physical thermodynamic method of holding at 80℃ for 10 minutes is used to kill the residual live bacteria inside the material; the vacuum drying of the fermented material needs to be carried out under specific parameters of 50℃ temperature and -0.09MPa vacuum; the standard for determining that the material has reached constant weight is set as the material weight change range ≤0.5% within 2 consecutive hours; and the final crushing and molding step requires that the powdered material must pass through an 80-mesh standard sieve.

[0012] Preferably, the content of catalpol in Rehmannia glutinosa fermented by the compound bacteria is quantitatively increased by 57.6% and the absolute content is ≥25.0mg / g. The macromolecules in the system are deconstructed and transformed, resulting in an increase of 126.4% in the content of soluble dietary fiber. The finished product test indicators meet the safety limit requirements for non-live bacteria products in the Chinese Pharmacopoeia and GB 4789.34-2016.

[0013] This invention provides a method for preparing and applying a compound bacterial fermentation of Rehmannia glutinosa with laxative function. It has the following beneficial effects: 1. This invention utilizes a synergistic fermentation system of four probiotics in a specific ratio of live bacteria. Lactobacillus acidophilus produces acid to promote the dissolution of catalpol, Bifidobacterium longum secretes enzymes to regulate the flora, Lactobacillus casei degrades insoluble cellulose, and Saccharomyces cerevisiae secretes proteases to activate intestinal peristalsis-related enzymes. This achieves a significant enhancement of bowel-regulating and laxative effects through multiple pathways, resulting in a 126.4% increase in soluble dietary fiber and a 57.6% increase in catalpol content.

[0014] 2. By employing safe bacterial strains in conjunction with a strict 80°C thermodynamic sterilization process during the fermentation termination period, this invention completely blocks the excessive metabolic behavior of harmful live bacteria during subsequent storage, achieving a high level of food safety with no risk of damage to the spleen and stomach mucosa under long-term continuous consumption.

[0015] 3. This invention achieves a simple and controllable process flow, clear definition of orthogonal parameters, and the ability to be rapidly transformed into large-scale industrial production of 50-100kg per batch by using a constant-temperature static fermentation at a mild temperature of 37℃ in combination with a fermentation tank and a vacuum drying oven. Attached Figure Description

[0016] Figure 1 This is a graph showing the dynamic changes of pH value and viable cell count over time during the simultaneous compound fermentation process of this invention. Figure 2 is a comparison of the characteristic peaks of catalpol in fermented and unfermented Rehmannia glutinosa in the embodiments of the present invention by HPLC. Figure 3This is a comparison image of HE staining of small intestinal tissue morphological sections used in animal experiments of this invention. Detailed Implementation

[0017] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. Example 1

[0018] This invention provides a method for preparing Rehmannia glutinosa fermented with compound bacteria and having a laxative function, comprising the following steps: S1. Strict physical parameter control is implemented during the raw material pretreatment stage. 100 kg of authentic Rehmannia glutinosa raw materials from Jiaozuo, Henan Province, free from mold and insect infestation, were selected. The materials were rinsed three times with flowing purified water to thoroughly remove surface dirt and sand. The washed materials were then fed into a fully automatic slicer and precisely sliced ​​into thin slices with a uniform thickness of 3-5 mm. These slices were then laid flat in a 50℃ forced-air drying oven for constant-temperature dehydration. An XM60 rapid moisture analyzer was used for real-time monitoring, and drying was stopped when the batch moisture content stabilized at 9.2%. Finally, an FW100 universal pulverizer was used to completely crush the powder, forcing it through a 60-mesh standard sieve, resulting in 92 kg of uniformly sized Rehmannia glutinosa powder.

[0019] S2. The preparation and formulation of the compound microbial agent strictly follow the principles of microecological balance. Four core fermentation strains were retrieved from the preservation center: *Lactobacillus acidophilus* CICC 6074, *Bifidobacterium longum* CICC 6188, *Lactobacillus casei* CICC 6096, and *Saccharomyces cerevisiae* CICC 31512. For the lactic acid bacteria, they were individually inoculated into MRS medium and activated anaerobically at 37°C for 18 hours, then transferred to a 10L fermenter for 24 hours of expansion culture. For *Saccharomyces cerevisiae*, they were inoculated into YPD medium and activated with shaking at 30°C for 24 hours, then transferred to a 10L fermenter for 36 hours of aerated expansion culture. Suspensions of each strain in the logarithmic growth phase were detected and screened, with an extraction concentration of 1.2 × 10⁻⁶. 9 30 L of Lactobacillus acidophilus solution with a concentration of 1.1 × 10⁻⁶ CFU / mL 9 20 L of Bifidobacterium longum solution with a concentration of 1.0 × 10⁻⁶ CFU / mL 9 20 L of Lactobacillus casei culture at a concentration of 1.3 × 10⁻⁶ CFU / mL and 10 Lactobacillus casei culture at a concentration of 1.3 × 10⁻⁶ CFU / mL. 910 L of yeast culture at CFU / mL. A total of 80 L of the above-mentioned individual culture solutions were aseptically mixed, and the viable cell ratio of the resulting compound culture was precisely controlled at 3:2:2:1, with the overall total viable cell count determined to be 5.2 × 10⁻⁶. 9 CFU / mL.

[0020] S3. Construction of fermentation substrate and simultaneous fermentation operation 92 kg of the prepared Rehmannia glutinosa powder was weighed and added into a 500 L BLBIO-50SJ type large-capacity fermenter. 460 kg of sterile purified water filtered through a 0.22 μm microporous membrane was injected to create a standardized substrate environment with a 1:5 (w / v) material-to-water ratio. A 1 mol / L standard NaOH solution was added dropwise to precisely adjust the pH of the substrate system to 6.5. The tank was sealed and high-temperature steam was introduced, maintaining a high pressure of 121 °C and 0.1 MPa for 15 minutes to achieve thorough sterilization. The cooling water circulation valve in the tank jacket was opened, and the internal substrate temperature was reduced to 37 °C at a forced cooling rate of 5 °C / min. 46 L of the prepared compound bacterial agent (equivalent to 5% of the total substrate weight, v / w) was injected through a sterile inoculation pipe. The sterile agitator inside the tank was started and run at 80 rpm for 5 minutes to ensure the bacteria were uniformly anchored on the surface of the Rehmannia glutinosa powder. The system was switched to a 37°C constant temperature static fermentation mode for 72 hours. The changes in pH and viable cell count over time during fermentation are as follows: Figure 1 As shown, during sampling and testing at 12-hour intervals, the pH value of the system remained stable in the acidic range of 4.1-4.2 from the middle to the end of fermentation, and the total viable count remained consistently high at 9.8 × 10⁻⁶. 9 Up to 1.0×10 10 CFU / mL range.

[0021] S4. The post-processing and crushing / molding steps determine the stability of the final product quality. At the end of the fermentation cycle, instantaneous high-temperature steam is directly introduced into the jacket of the fermenter, causing the material temperature to rise rapidly to 80℃ and be kept constant for 10 minutes, completely blocking and killing all active residual bacteria. The paste-like fermentation product is then pumped into a DZF-6050 vacuum drying oven. The equipment operating parameters are set as follows: drying temperature 50℃, ultimate vacuum degree -0.09MPa. The material is continuously dehydrated in this vacuum negative pressure environment until it reaches an absolute constant weight state with a weight change of ≤0.5% for 2 consecutive hours (final moisture content measured as 8.5%). The dried agglomerates are then removed and subjected to secondary mechanical pulverization, and sieved through an 80-mesh standard sieve (sieve aperture size 0.18mm). The final yield of this batch was 68.5kg of simultaneously compound fermented Rehmannia glutinosa powder, with an overall process yield of 74.5%.

[0022] To provide rigorous data reference, a parallel control sample of unfermented Rehmannia glutinosa powder was prepared simultaneously. 10 kg of dried Rehmannia glutinosa powder from the same batch was extracted, and the substrate was treated strictly according to the above-mentioned 1:5 material-to-water ratio and autoclaving procedure, refraining from inoculating any exogenous microbial agents throughout the process. After undergoing identical vacuum drying, pulverizing, and sieving processes, 7.2 kg of unfermented Rehmannia glutinosa powder control sample was obtained.

[0023] Example 2: This example provides a comparative detection and analysis of the core components of fermentation products. The batch sample of the compound fermented powder produced in Example 1 was retrieved and compared with the control sample of the unfermented powder by the testing team for specific component comparison.

[0024] The content of catalpol was determined using an Agilent 1260 HPLC system equipped with a Diamonsil C18 column. The mobile phase was set to methanol-water (15:85, v / v), the flow rate was 1.0 mL / min, the column temperature was 30℃, and the detection wavelength was 210 nm. Soluble dietary fiber was determined according to the standard procedure of enzyme gravimetric method in GB 5009.88-2014. The pH value of the system was determined using a water extraction and settling method combined with a precision pH meter.

[0025] Figure 2 shows the HPLC chromatographic comparison of the characteristic peaks of catalpol in fermented and unfermented Rehmannia glutinosa after three independent parallel analyses. Figure 2a The image shows the HPLC chromatogram of the characteristic peak of catalpol in unfermented Rehmannia glutinosa. Figure 2b The HPLC chromatogram shows the characteristic peak of catalpol in fermented Rehmannia glutinosa. Data confirms that the catalpol content in the unfermented sample was 18.25±0.34 mg / g, while the content in the fermented sample increased significantly to 28.76±0.51 mg / g, a relative increase of 57.6%. The soluble dietary fiber content in the unfermented sample was 12.50±0.25 mg / g, while the fermented sample saw a sharp increase to 28.30±0.40 mg / g, an absolute increase of 126.4%. Simultaneously, the pH of the fermentation system decreased significantly from the initial 6.2±0.1 to 4.1±0.2. The detection data fully demonstrate that the microbial metabolic process not only releases bound active glycosides but also decomposes a large amount of insoluble cellulose backbone into soluble oligosaccharides with laxative effects.

[0026] Example 3: Animal model evidence of the laxative effect of fermented Rehmannia glutinosa A mouse model of constipation was established. The experimental animals were randomly divided into a blank control group (saline gavage), an unfermented Rehmannia glutinosa group (1.0 g / kg gavage), and a fermented Rehmannia glutinosa group (1.0 g / kg gavage), and were treated with the drug for 30 consecutive days.

[0027] Defecation behavior tracking results showed that the time to first black stool in the control group was 45.2±5.8 min, in the unfermented group it was 38.5±4.2 min, while the mice in the fermented Rehmannia group excreted black stool within 28.3±3.6 min. In 24-hour fecal pellet monitoring, the fermented Rehmannia group produced a high number of 35.6±4.1 pellets, significantly exceeding the 18.5±3.2 pellets in the control group and the 25.3±3.8 pellets in the unfermented group. The Bristol score for feces showed that the fermented Rehmannia group scored 5.1±0.5, exhibiting a moist and formed fecal morphology characteristic of good health.

[0028] Anatomical experiments on the propulsion rate of charcoal in the small intestine further revealed the underlying mechanism. The propulsion rate in the control group was only 58.3±6.1%, while in the fermented rehmannia group, the propulsion rate of charcoal in the small intestine jumped to 78.4±4.8%. Microscopic examination of the sections showed... Figure 3 As shown, the intestinal villi length of the fermentation group increased significantly from 320±25μm in the control group to 450±35μm, and the villi were arranged densely and completely, thus completely eliminating the potential risk of gastrointestinal irritation and damage from traditional Rehmannia glutinosa components.

[0029] Example 4: Preparation of specific dosage forms for laxative and bowel-regulating foods and medicines Based on the standard fermentation powder produced in Example 1, it can be easily converted into the following dosage forms to meet the needs of different market terminals: Solid tableting process: Weigh 80% by weight of fermented Rehmannia glutinosa powder as the main ingredient. Add 15% microcrystalline cellulose as a plastic filler and 5% magnesium stearate as a release lubricant. The material is continuously tumbled in a three-dimensional mixing device until it reaches a homogeneous state. Start a high-speed rotary tableting machine and control the die pressure to compress the mixed powder into hard tablets weighing 0.3g each, each tablet precisely loaded with 0.24g of the fermented core ingredients, suitable for portable consumption.

[0030] Capsule filling procedure: Weigh out 80% of the fermented powder by mass and evenly mix in 20% pharmaceutical-grade high-quality lactose powder to adjust the flowability. Add the uniformly mixed powder into the storage hopper of the fully automatic capsule filling machine, and continuously fill it into standard No. 0 medical gelatin hollow capsule shells according to the filling parameters of 0.3g net weight per capsule, to achieve high-density single-capsule encapsulation of 0.24g fermented Rehmannia glutinosa.

[0031] Instant granule forming process: 70% fermented main ingredient powder is used, with 25% sucrose powder and 5% dextrin added to adjust the texture and granulation viscosity. A wet granulation machine is started to prepare a soft mass, which is then extruded into granules through a screen with a specific aperture. The wet granules are pushed into a fluidized bed dryer for rapid drying until the internal moisture content is ≤5%. High-precision packaging is performed using an automatic packaging machine; each 3g bag contains 2.1g of fermented rehmannia root and dissolves very easily when brewed.

[0032] Oral liquid preparation route: Extract 10% fermented powder, mix with 5% sucrose granules, and pour into 85% purified water. Activate the jacketed heating device to maintain the water temperature at 80℃ to accelerate the complete dissolution and hydration of the solid material. The coarsely filtered liquid is forced into a 0.22μm microporous membrane to intercept minute impurities. After undergoing a high-temperature sterilization process at 121℃ for 15 minutes, the clear filtrate is dispensed into ampoules under a sterile Class 100 laminar flow hood. Each 10mL ampoule contains 1.0g of fermented Rehmannia glutinosa extract.

[0033] Powder packaging method: 83.3% high-concentration fermented powder is extracted and directly combined with 16.7% maltodextrin as a dispersion carrier. The mixture is thoroughly mixed in a gravity-free biaxial paddle mixer. Aluminum foil composite film is used for sealing and packaging using specialized powder packaging equipment. Each bag is set to weigh 3g, ensuring a net content of up to 2.5g of fermented rehmannia root per bag, suitable for high-dose oral administration in patients with severe irritable bowel syndrome.

[0034] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A method for preparing Rehmannia glutinosa by simultaneous fermentation with compound bacteria and laxative function, characterized in that, Includes the following steps: S1. Extract Lactobacillus acidophilus CICC 6074, Bifidobacterium longum CICC 6188, Lactobacillus casei CICC6096 and Saccharomyces cerevisiae CICC 31512 at a preset concentration, and aseptically mix the bacterial suspensions of the above four microorganisms in a ratio of 3:2:2:1 to construct a compound microbial agent. S2. Take Rehmannia glutinosa powder as the substrate raw material, add sterile purified water according to the predetermined feed water ratio, adjust the pH of the mixed system and then perform high-temperature steam sterilization. After the system is cooled to the appropriate inoculation temperature, the fermentation substrate is obtained. S3. The aforementioned compound microbial agent is added to the fermentation substrate according to the volume-mass ratio. After being thoroughly mixed, the mixture is subjected to static synchronous compound fermentation under constant temperature conditions. S4. After the fermentation cycle is completed, the microbial metabolic process is completely terminated by high-temperature insulation. The resulting material is then transferred to a vacuum environment for low-temperature drying until it reaches a constant weight. Finally, it is pulverized and sieved to obtain synchronous compound fermented Rehmannia glutinosa powder.

2. The method for preparing Rehmannia glutinosa with compound bacteria and laxative function according to claim 1, characterized in that, The total viable bacteria count in the compound microbial agent reaches 5 × 10⁶. 9 For strains at CFU / mL or higher, each individual bacterial species constituting the microbial agent must undergo activation and expansion culture procedures beforehand. Lactic acid bacteria are activated on MRS medium in an anaerobic environment at 37°C for 18 hours and expanded culture for 24 hours, respectively. Saccharomyces cerevisiae is activated on YPD medium at 30°C with shaking for 24 hours and expanded culture with aeration for 36 hours. The concentration of each individual bacterial suspension is uniformly adjusted to 1×10⁻⁶ before mixing. 9 CFU / mL.

3. The method for preparing Rehmannia glutinosa with compound bacteria and laxative function according to claim 1, characterized in that, The specific processing procedure for the Rehmannia glutinosa powder is as follows: Select Rehmannia glutinosa from Jiaozuo, Henan Province, that is free from mold and insect infestation. Rinse it three times with flowing purified water to remove impurities. Then cut it into thin slices with a thickness of 3-5 mm. Place the slices in a 50°C forced-air drying oven to dry until the moisture content is less than 10% as measured by a moisture analyzer. Use mechanical pulverizing equipment to pulverize the dried material and make it pass completely through a 60-mesh standard sieve.

4. The method for preparing Rehmannia glutinosa with compound bacteria and laxative function according to claim 1, characterized in that, During the preparation of the fermentation substrate, the ratio of Rehmannia glutinosa powder to sterile purified water was set at 1:5 (w / v). HCl or NaOH solution with a concentration of 1 mol / L was added dropwise to adjust the pH of the substrate system to the range of 6.5±0.

1. The sterilization program was carried out at 121℃ and 0.1MPa high-pressure steam for 15 minutes. After the sterilization was completed, the temperature was quickly reduced to 37℃ by the cooling water circulation system at a cooling rate of 5℃ / min.

5. The method for preparing Rehmannia glutinosa with a compound bacteria fermentation function according to any one of claims 1, characterized in that, The inoculum amount of the compound microbial agent is 5% (v / w) of the total weight of the fermentation substrate. After inoculation, a sterile agitator should be started and continuously stirred at 80 rpm for 5 minutes to ensure uniform dispersion. The entire fermentation process is maintained at a constant temperature of 37℃ for up to 72 hours. During this period, samples should be taken every 12 hours to confirm that the pH value of the system is maintained at 4.1±0.2 and the overall viable cell count is maintained at ≥9×10⁻⁶. 9 Above the standard of CFU / mL.

6. The method for preparing Rehmannia glutinosa with compound bacteria and laxative function according to claim 1, characterized in that, The specific process conditions for the fermentation termination and post-processing stages are as follows: The physical thermodynamic method of holding at 80℃ for 10 minutes is used to kill the residual live bacteria inside the material. The vacuum drying of the fermented material must be carried out under specific parameters of 50℃ temperature and -0.09MPa vacuum. The standard for determining that the material has reached constant weight is set as the material weight change range ≤0.5% within 2 consecutive hours. The final crushing and molding step requires that the powdered material must pass through an 80-mesh standard sieve.

7. The compound microbial simultaneous fermentation of Rehmannia glutinosa prepared by the preparation method according to any one of claims 1-6, characterized in that, Compared to the same raw materials without fermentation, the content of catalpol in Rehmannia glutinosa fermented by compound bacteria was quantitatively increased by 57.6% and the absolute content was ≥25.0mg / g. The macromolecules in the system were deconstructed and transformed, resulting in an increase of 126.4% in soluble dietary fiber content. The finished product test indicators met the safety limits for non-live bacteria products in the Chinese Pharmacopoeia and GB 4789.34-2016.

8. The application of the compound bacteria-fermented Rehmannia glutinosa as described in claim 7 in the preparation of laxative products for constipation caused by intestinal dryness and fluid deficiency, characterized in that... The products cover the fields of functional foods, health foods, and pharmaceutical manufacturing. The specific pharmaceutical dosage forms for which the active ingredient is applicable include tablets, capsules, granules, oral liquids, or powders.

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