Gastric-acid-resistant high-activity coprophilous fungus transplantation freeze-dried powder and preparation method thereof

Abstract

The invention discloses gastric-acid-resistant high-activity coprophilous fungus transplantation freeze-dried powder and a preparation method, and belongs to the technical field of medicines.The preparation method comprises the steps that coprophilous fungus sludge and a protective agent are mixed according to the mass ratio of 1: (1-2), and freeze drying is conducted to prepare the gastric-acid-resistant high-activity coprophilous fungus transplantation freeze-dried powder; wherein the protective agent comprises the following raw materials in percentage by mass: 2%-6% of trehalose, 5%-10% of resistant dextrin, 5%-8% of skimmed milk, 3%-8% of vitamin C, 1%-5% of galactooligosaccharide, 0.8%-3% of glycerol, 0.05%-0.5% of lactase and the balance of water; the invention solves the problems of low survival rate of coprophilous fungus transplanted viable bacterial powder, low bacterial count content per unit bacterial powder and poor gastric acid resistance of the existing freeze-dried coprophilous fungus transplanted viable bacterial powder.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and in particular to a gastric acid-resistant high-activity fecal bacteria transplantation freeze-dried powder and a preparation method thereof. Background Art

[0002] Fecal Microbiota Transplantation (FMT), as an innovative therapy, is gradually showing its unique potential and value in the medical field. This therapy carefully screens fecal samples from healthy individuals, extracts functional flora from them, and uses advanced biotechnology to safely and effectively transplant these flora into the patient's intestines. This process aims to reshape the patient's intestinal microecological balance and restore flora diversity, thereby treating a variety of diseases caused by intestinal flora imbalance, such as Clostridium difficile infection, inflammatory bowel disease, and some metabolic diseases. The successful practice of FMT not only reveals the close connection between intestinal microorganisms and health, but also opens up new paths for exploring new treatment strategies.

[0003] However, in practical applications, a major technical challenge facing fecal microbiota transplantation is how to efficiently and stably preserve these complex and sensitive microbial communities. Vacuum freeze-drying technology, as a widely used method for microbial preservation, can effectively remove moisture from samples and prolong the storage time, but its applicability to fecal microbiota communities has many limitations. Fecal microbiota communities are composed of hundreds of different microorganisms, each of which has unique physiological characteristics and environmental adaptability, especially high anaerobicity and sensitivity to environmental conditions, which makes it easy to cause cell structure damage, loss of activity and other problems during the freeze-drying process. The rapid removal of moisture and temperature fluctuations may cause irreversible damage to key structures such as microbial cell walls and membrane proteins, thereby affecting the overall function and therapeutic effect of the microbial community. Therefore, how to maximize the protection of the activity, structure and functional integrity of fecal microbiota communities during the freeze-drying process has become a key bottleneck restricting the widespread application of FMT technology.

[0004] In order to reduce the damage of freeze-drying to microorganisms, specific protective agents (such as sugars, starches, proteins, etc.) are needed. They can effectively prevent cell wall rupture, membrane protein denaturation and other structural damage, thereby ensuring that the flora maintains long-term activity and stability after drying. In addition, the strong acid environment in the stomach poses a great challenge to the survival of the flora. Some biomacromolecules not only serve as freeze-drying protective agents, but also have excellent gastric acid resistance. With the increasing importance of intestinal flora in health management, how to efficiently preserve and stably produce flora has become a core technical problem in promoting the industrialization of flora. To solve this problem, it is necessary not only to improve the freeze-drying process, but also to achieve systematic breakthroughs from the research and development of protective agents to the maintenance of flora proliferation, thereby laying the foundation for the development and application of flora transplantation related products. Summary of the invention

[0005] The present invention provides a gastric acid-resistant high-activity fecal microbiota transplantation freeze-dried powder and a preparation method thereof, which solves the problems of low bacterial survival rate, low bacterial count per unit bacterial powder and poor gastric acid resistance of the existing freeze-dried fecal microbiota transplantation live bacterial powder production.

[0006] To achieve the above-mentioned purpose, the present invention provides the following technical scheme: a freeze-dried protective agent for live bacteria in fecal microbiota transplantation, wherein the raw materials of the protective agent include, by mass percentage, trehalose 2%~6%, resistant dextrin 5%~10%, skim milk 5%~8%, vitamin C 3%~8%, oligosaccharide 1%~5%, glycerol 0.8%~3%, lactase 0.05%~0.5%, and the balance is water.

[0007] Furthermore, the raw materials of the protective agent include, by mass percentage, trehalose 6%, resistant dextrin 8%, skim milk 4%, vitamin C 3%, galacto-oligosaccharide 2%, glycerol 2%, lactase 0.3%, and the balance is water.

[0008] The present invention also provides a method for preparing the above-mentioned fecal microbiota transplant live bacteria freeze-dried protective agent, the specific steps of which are as follows: Obtain filter-sterilized vitamin C solution; Obtain a mixture of high temperature sterilized trehalose, resistant dextrin, skim milk, galacto-oligosaccharide and glycerol; The vitamin C solution and the mixed solution are mixed and then lactase is added to obtain a freeze-dried protective agent for live bacteria in fecal microbiota transplantation.

[0009] Furthermore, in the step of obtaining the filter-sterilized vitamin C solution, the filter membrane has a pore size of 0.22 μm.

[0010] Furthermore, in the step of obtaining a high-temperature sterilized mixed solution of trehalose, resistant dextrin, skim milk, galacto-oligosaccharide and glycerol, the high-temperature sterilization condition is 115° C. for 10 min to 15 min.

[0011] The present invention also provides a gastric acid-resistant and highly active fecal microbiota transplantation freeze-dried powder, comprising the above-mentioned fecal microbiota transplantation live bacteria freeze-dried protective agent.

[0012] Furthermore, the fecal bacteria sludge is mixed with the protective agent in a mass ratio of 1:1 to 2, and freeze-dried to obtain gastric acid-resistant and highly active fecal bacteria transplantation freeze-dried powder.

[0013] Furthermore, the freeze-drying conditions are specifically as follows: pre-freezing at -80°C to -50°C for 1h to 4h, and then gradient sublimation, and the program setting parameters are: under 0Pa to 5Pa conditions, running at -20°C to -10°C for 1h to 2h; under 5Pa to 10Pa conditions, running at -10°C to 0°C for 1h to 2h; under 10Pa to 15Pa conditions, running at 5°C to 10°C for 8h to 12h; and then freeze-drying at 20°C to 25°C for 3h to 8h under 5Pa to 25Pa conditions.

[0014] The present invention also provides a method for improving the survival rate of freeze-dried powder of fecal microbiota transplantation, wherein the fecal microbiota sludge and the above-mentioned protective agent are stirred and mixed at a mass ratio of 1:1 to 2 for 10 to 15 minutes, and then freeze-dried.

[0015] The present invention also provides a method for improving the gastric acid resistance of fecal microbiota transplant freeze-dried powder, wherein the fecal microbiota sludge and the above-mentioned protective agent are stirred and mixed at a mass ratio of 1:1 to 2 for 10 minutes to 15 minutes, and then freeze-dried.

[0016] Compared with the prior art, the present invention has at least the following beneficial effects: The present invention provides a fecal bacteria transplant live bacteria freeze-dried protective agent, the freeze-dried protective agent components and ratios of which have been screened and optimized through a large number of experiments, and can significantly reduce the damage to the bacteria during vacuum freeze drying, effectively maintain the integrity and activity of the bacteria, and thus improve the survival rate of the fecal bacteria. The freeze-dried protective agent components of the present invention are all pharmaceutical or food grade raw materials, and are widely available and low in cost. The preparation and use methods of the protective agent of the present invention are simple, highly operable, and suitable for standardized industrial production.

[0017] The present invention also provides a gastric acid-resistant and high-activity fecal bacteria transplantation freeze-dried powder, which reduces the damage to the bacteria during the vacuum freeze-drying process through a freeze-drying protective agent, thereby improving the survival rate of the fecal bacteria. In addition, based on the optimal protective agent formula of the above-mentioned gastric acid-resistant and high-activity fecal bacteria transplantation freeze-dried powder, a set of gradient sublimation freeze-drying procedures is optimized, which shortens the freeze-drying time and reduces the loss of live bacteria during the freeze-drying process. At the same time, the moisture content of the freeze-dried bacterial powder is controlled to not exceed 5% (consistent with the corporate standard level of probiotic powder on the market), which is beneficial to maintaining the stability of the bacteria during the storage period, extending the shelf life, and greatly improving the production efficiency, thereby achieving the dual goals of reducing costs and increasing efficiency.

[0018] The freeze-dried bacterial powder prepared by the invention has a loose texture and is convenient to crush, the protective agent components are non-toxic and harmless, the number of live bacteria in the freeze-dried bacterial powder is high, the later stability is good, and the survival rate is more than 80% when stored at -20°C for one year, which meets the requirements of drug production, and the obtained bacterial powder can be directly applied to the field of microecological medicine, and has huge market potential. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1The effect of storage time on the survival rate of different bacterial powders. DETAILED DESCRIPTION

[0020] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments.

[0021] The first aspect of the present invention provides a fecal microbiota transplant live bacteria freeze-dried protective agent, comprising the following ingredients in percentage by mass: trehalose 2%~6%, resistant dextrin 5%~10%, skim milk 5%~8%, vitamin C 3%~8%, galacto-oligosaccharide 1%~5%, glycerol 0.8%~3%, lactase 0.05%~0.5%, and the balance is water.

[0022] Preferably, the source of lactase is commercial lactase from Henan Wanbang Industrial, specifically a compound lactase synthesized by microorganisms with an enzyme activity of 50,000 U. The addition of lactase can improve the digestion and absorption rate of skim milk, and the resulting fecal microbiota transplant live bacteria freeze-dried protective agent is suitable for people with lactose intolerance.

[0023] More preferably, the mass percentage of lactase is 0.1% to 0.3%.

[0024] Preferably, the fecal microbiota transplant live bacteria freeze-dried protective agent contains the following ingredients in percentage by mass: trehalose 6%, resistant dextrin 8%, skim milk 4%, vitamin C 3%, galacto-oligosaccharide 2%, glycerol 2%, lactase 0.3%, and the balance is water.

[0025] The second aspect of the present invention provides a method for preparing the above-mentioned fecal microbiota transplant live bacteria freeze-dried protective agent, comprising the following steps: Step 1: Dissolve vitamin C in an appropriate amount of water, and filter and sterilize with a filter membrane to obtain solution A; Step 2: dissolve trehalose, resistant dextrin, skim milk, galacto-oligosaccharide and glycerol with the remaining amount of water, fully dissolve and mix, and then sterilize to obtain solution B; Step 3: Mix solution A and solution B evenly and add lactase to obtain a complete protective agent, which is stored at 4°C for later use.

[0026] Preferably, the pore size of the filter membrane in step 1 is 0.22 μm filter membrane; Preferably, the sterilization condition in step 2 is 115° C. for 10 min to 15 min.

[0027] The third aspect of the present invention provides a method for using the above-mentioned fecal microbiota transplant live bacteria freeze-dried protective agent, comprising the following steps: Step 1. Stir the bacterial sludge collected by centrifugation and the above-mentioned fecal microbiota transplant live bacteria freeze-dried protective agent in a mass ratio of 1:1-2 for 10min-15min, place it in a freeze dryer, pre-freeze at -80℃~-50℃ for 1h~4h, and then perform gradient sublimation. The program setting parameters are: under 0Pa~5Pa conditions, run at -20℃~-10℃ for 1h~2h; under 5~10Pa conditions, run at -10℃~0℃ for 1h~2h; under 10Pa~15Pa conditions, run at 5℃~10℃ for 8h~12h; then freeze-dry at 20℃~25℃ for 3h~8h under 5Pa~25Pa conditions; the fecal microbiota transplant live bacteria freeze-dried powder can be obtained.

[0028] The freeze-dried protective agent of the present invention can stabilize the survival rate of fecal bacteria at more than 85%, and the number of live bacteria in the bacterial powder is 6×10 11 CFU / g, the survival rate of bacterial powder is more than 80% when stored for one year, which greatly improves the freeze-dried survival rate of bacteria and the stability of bacterial powder in the later period.

[0029] The present invention is further described in detail below in conjunction with examples, but the embodiments of the present invention are not limited thereto. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art.

[0030] Example 1 Effect of lyophilization protectant on the survival rate of bacterial powder before and after freeze drying In this example, composite freeze-drying protective agents composed of different added amounts of trehalose, resistant dextrin, skim milk, vitamin C, oligosaccharides, glycerol, and lactase were respectively prepared and used according to the protective agent preparation and use methods of the present invention. After being mixed with bacterial mud, freeze-drying was performed under the same conditions, and each factor was taken at 3 levels (Table 1). The freeze-drying survival rate of the bacteria was further calculated to evaluate the effect of freeze-drying protection.

[0031] Freeze-dried survival rate = [(number of live bacteria per freeze-dried powder × mass of freeze-dried powder) / (number of live bacteria per mud × mass of mud)] × 100% Table 1. Experimental factor levels for optimizing the formula of live bacteria composite freeze-dried protective agent for fecal microbiota transplantation

[0032] Table 2 Results of orthogonal test for optimization of formula of live bacteria composite freeze-dried protective agent for fecal microbiota transplantation

[0033] As shown in Table 2, the primary and secondary factors affecting the survival rate of freeze-dried bacteria are the addition amount of vitamin C, resistant dextrin, lactase, trehalose, galacto-oligosaccharide, skim milk, and glycerol, that is, D>B>G>A>E>C>F. And the optimization scheme can be obtained as A3B2C1D3E1F2G3, that is, trehalose 6%, resistant dextrin 8%, skim milk 4%, vitamin C 3%, galacto-oligosaccharide 2%, glycerol 2%, and lactase 0.3%. Through the verification experiment of the optimization scheme A3B2C1D3E1F2G3, the survival rate of freeze-dried bacterial powder obtained reached 85%, and the protection effect of this freeze-dried protective agent formula was the best.

[0034] Example 2 Effect of freeze-drying procedure on the survival rate of freeze-dried bacteria After the fecal bacteria dilution was squeezed and filtered, the filtrate was centrifuged to collect the bacterial sludge. The bacterial sludge collected by centrifugation was sampled and the number of viable bacteria was detected by PMA-qPCR method. 30g of bacterial sludge was mixed with the optimized formula of freeze-drying protective agent at a mass ratio of 1:1 for 10 minutes, and placed in a vacuum freeze dryer. The freeze-drying program was set according to Table 3 for each experimental group to obtain freeze-dried powder of live bacteria for fecal bacteria transplantation in each group. The mass of the bacterial powder was weighed and the number of viable bacteria was sampled and tested, and the freeze-drying survival rate was further calculated.

[0035] Table 3 Effect of freeze-drying procedure on the survival rate of freeze-dried bacteria

[0036] During the freeze-drying process, key parameters such as sublimation temperature, sublimation time, and decomposition time were optimized to ensure the lowest moisture content and shortest freeze-drying time of the freeze-dried product and improve production efficiency. From the above results, it can be seen that the freeze-drying procedure of experimental group 3 took a shorter time than other experimental groups, had the highest survival rate, and the moisture content of the freeze-dried bacterial powder was only 3.2%, which ensured the stress resistance of the bacterial powder and was conducive to extending the storage period.

[0037] Example 3 Effect of freeze-drying protective agent on the survival rate of bacterial powder during storage From the results of the extreme difference analysis of Example 1, it can be obtained that glycerol, trehalose, skim milk, and galacto-oligosaccharides are secondary factors that affect the freeze-dried survival rate during the vacuum freeze-drying process. Therefore, their synergistic protective effects as permeability protectants, non-permeability protectants, protein protectants, and prebiotics in low-temperature frozen storage were further verified. Take 30g of bacterial mud and the freeze-dried protective agent of each experimental group (as shown in Table 4) at a mass ratio of 1:1 and stir for 10 minutes. Place them in a vacuum freeze dryer for freeze drying. Take 10g of the obtained freeze-dried bacterial powder and fill capsules. Each capsule is filled with 0.2g~0.4g of bacterial powder. The filled capsules are stored at -20°C. Take one capsule from each group at 1, 2, 3, 6, 9, and 12 months to detect the number of live bacteria in the bacterial powder, and further calculate the live bacteria survival rate during the storage period. The results are shown in Table 1. Figure 1 .

[0038] Table 4 Effect of freeze-drying protective agent on the survival rate of bacterial powder during storage

[0039] pass Figure 1 It can be seen that the survival rate of freeze-dried bacterial powder in experimental group 1 was only 16.7% after storage for 12 months, while the survival rates of experimental groups 2, 3, 4, and 5 with the addition of "permeable protectant", "permeable protectant + non-permeable protectant", "permeable protectant + non-permeable protectant + protein protectant", and "permeable protectant + non-permeable protectant + protein protectant + prebiotics" were 36.1%, 56.8%, 77.4%, and 91.5%, respectively, which were significantly higher than those in experimental group 1, indicating that the added protectant can increase the storage stability of bacterial powder and extend the shelf life. Among them, the combination of glycerol, trehalose, skim milk, and oligosaccharides can play a synergistic protective role of permeable protectants, non-permeable protectants, protein protectants, and prebiotics. When stored at -20°C for 12 months, the survival rate of bacterial powder cells was 91.5%.

[0040] It can be seen that the freeze-dried protective agent of the present invention can significantly improve the survival rate of the fecal microbiota transplantation freeze-dried powder, ensuring that the product maintains a sufficient number of live bacteria during storage and use. In addition, by optimizing the protective agent formula and preparation process, the production cost is reduced, and the cost-effectiveness and market competitiveness of the product are improved.

[0041] Example 4 Gastric Acid Resistance of Freeze-dried Bacterial Powder By gradually adding irritating acidic substances to acclimate the bacterial flora, adaptive changes can be induced in the bacterial flora, enhancing cell membrane stability and the production of resistance metabolites, thereby improving the tolerance of the bacterial flora to external environmental pressure. This method is widely used in the industrial, environmental and food fields. Therefore, the treatment time of vitamin C acclimatization of the bacterial flora is further optimized to obtain the highest survival rate of the bacterial flora after passing through the gastrointestinal tract.

[0042] After the fecal bacteria dilution was squeezed and filtered, the filtrate was centrifuged to collect the bacterial sludge. The bacterial sludge collected by centrifugation was sampled and the number of viable bacteria was detected by PMA-qPCR method. 30g of bacterial sludge was mixed with the optimized formula of freeze-dried protective agent at a mass ratio of 1:1, and treated for 0min, 5min, 10min, and 15min (0min was obtained by mixing the bacterial sludge with the optimized formula of freeze-dried protective agent without vitamin C), and then placed in a vacuum freeze dryer. After freeze drying, the freeze-dried powder of live bacteria for fecal bacteria transplantation in each group was obtained, and the mass of the bacterial powder was weighed and sampled for the number of viable bacteria.

[0043] Simulated gastric digestive fluid (SGF) was prepared as follows: 0.1 g NaCl, 350 μL 0.1 mol·L-1 HCl were dissolved in 50 mL water, pH was adjusted to 2.0, and 2% (w / v) pepsin was contained. Then, the freeze-dried bacterial powder of each experimental group and the corresponding bacterial powder enteric-coated capsules of each group were added to 9 mL SGF, placed in an anaerobic chamber at 37°C and 120 r / min in a shaker to simulate gastric digestion, and taken out after 2 h. After the freeze-dried bacterial powder was digested with gastric juice, 500 μL of digestive fluid was aspirated, centrifuged at 12000 rpm for 2 min, and the bacterial sludge sediment was immediately tested for the total number of viable bacteria by PMA-qPCR. 0.1 g of the contents of the bacterial powder enteric-coated capsules were taken and the total number of viable bacteria was immediately tested by PMA-qPCR.

[0044] Gastric dissolution survival rate = (total number of live bacteria after gastric dissolution / total number of live bacteria before gastric dissolution) × 100% Table 5 Changes in the number of live bacteria per unit before and after simulated gastric digestion

[0045] The results of gastric acid resistance of freeze-dried bacterial powder are shown in Table 5. The gastric survival rate of freeze-dried bacterial powder in different treatment groups containing the optimized formula of vitamin C freeze-dried protective agent directly contacted with SGF for 2 hours and digested was not significantly different from the gastric survival rate of its bacterial powder enteric-coated capsules, indicating that the bacterial flora after domestication with the optimized formula of vitamin C freeze-dried protective agent of the present invention has the ability to resist gastric acid, and its gastric acid resistance is equivalent to that of commercial enteric-coated capsules. The results show that the freeze-dried survival rate and gastric survival rate obtained when the vitamin C domestication treatment time is 10 minutes are significantly higher than those of other treatment groups.

[0046] It can be seen that the lyophilized protective agent of the present invention can significantly improve the gastric acid resistance of the fecal microbiota transplantation freeze-dried powder, ensuring that the bacterial powder can maintain a sufficient number of live bacteria when passing through the gastric acid environment, thereby exerting the expected microecological treatment effect. By enhancing the gastric acid resistance of the bacterial powder, this method effectively guarantees the therapeutic effect of fecal microbiota transplantation, improves patient satisfaction and the stability of the therapeutic effect. Moreover, the method of the present invention is simple and clear, easy to operate and implement, and is suitable for production and application scenarios of various scales, and has broad market prospects and promotion value.

[0047] Therefore, the protective agent provided by the present invention can not only play an overall synergistic role during vacuum freeze-drying and low-temperature frozen storage to improve the survival rate and stability of bacterial powder, but also has a significant effect in resisting gastric acid, greatly improving the gastric acid resistance of live bacteria, which is conducive to the development and application of oral flora transplantation related products.

Claims

1. A fecal microbiota transplant live bacteria freeze-dried protective agent, characterized in that: Calculated by mass percentage, the raw materials of the protective agent include: 2%-6% trehalose, 5%-10% resistant dextrin, 5%-8% skim milk, 3%-8% vitamin C, 1%-5% galacto-oligosaccharide, 0.8%-3% glycerol, 0.05%-0.5% lactase, and the balance is water.

2. The fecal microbiota transplant live bacteria freeze-dried protective agent according to claim 1, characterized in that: Calculated by mass percentage, the raw materials of the protective agent include: 6% trehalose, 8% resistant dextrin, 4% skim milk, 3% vitamin C, 2% galacto-oligosaccharide, 2% glycerol, 0.3% lactase, and the balance is water.

3. The method for preparing a fecal microbiota transplant live bacteria freeze-dried protective agent according to claim 1 or 2, characterized in that: The specific steps are as follows: Obtain filter-sterilized vitamin C solution; Obtain a mixture of high temperature sterilized trehalose, resistant dextrin, skim milk, galacto-oligosaccharide and glycerol; The vitamin C solution and the mixed solution are mixed and then lactase is added to obtain a freeze-dried protective agent for live bacteria in fecal microbiota transplantation.

4. The method for preparing a fecal microbiota transplant live bacteria freeze-dried protective agent according to claim 3, characterized in that: In the step of obtaining the filter-sterilized vitamin C solution, the filter membrane used is a filter membrane with a pore size of 0.22 μm.

5. The method for preparing a fecal microbiota transplant live bacteria freeze-dried protective agent according to claim 3, characterized in that: In the step of obtaining a high-temperature sterilized mixed solution of trehalose, resistant dextrin, skim milk, galacto-oligosaccharide and glycerol, the high-temperature sterilization condition is 115° C. for 10 min to 15 min.

6. A gastric acid-resistant and highly active fecal microbiota transplantation freeze-dried powder, characterized in that: It includes the fecal microbiota transplant live bacteria freeze-dried protective agent according to claim 1 or 2.

7. The gastric acid-resistant and highly active fecal bacteria transplantation freeze-dried powder according to claim 6, characterized in that: The fecal bacteria sludge is mixed with the protective agent in a mass ratio of 1:1 to 2, and freeze-dried to obtain gastric acid-resistant and highly active fecal bacteria transplant freeze-dried powder.

8. The gastric acid-resistant and highly active fecal bacteria transplantation freeze-dried powder according to claim 7, characterized in that: The freeze-drying conditions are specifically as follows: pre-freeze at -80°C to -50°C for 1h to 4h, and then perform gradient sublimation. The program setting parameters are as follows: under 0Pa to 5Pa conditions, run at -20°C to -10°C for 1h to 2h; under 5Pa to 10Pa conditions, run at -10°C to 0°C for 1h to 2h; under 10Pa to 15Pa conditions, run at 5°C to 10°C for 8h to 12h; and then freeze-dry at 20°C to 25°C for 3h to 8h under 5Pa to 25Pa conditions.

9. A method for improving the survival rate of fecal microbiota transplantation freeze-dried powder, characterized in that: The fecal fungus sludge and the protective agent according to claim 1 or 2 are stirred and mixed at a mass ratio of 1:1 to 2 for 10 to 15 minutes, and then freeze-dried.

10. A method for improving the gastric acid resistance of fecal microbiota transplantation freeze-dried powder, characterized in that: The fecal fungus sludge and the protective agent according to claim 1 or 2 are stirred and mixed at a mass ratio of 1:1 to 2 for 10 to 15 minutes, and then freeze-dried.

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