Microbial flora extraction and recombination fermentation application method in vegetable fermentation liquor
By extracting microbial flora from the fermentation broth of different vegetables, preparing special fermentation agents and performing recombinant fermentation, the problems of low utilization rate of vegetable fermentation broth and lack of special fermentation agents in the market are solved, and efficient and excellent fermentation effect is achieved.
Patent Information
- Application Number
- CN202510108857.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-01-23
- Publication Date
- 2025-04-11
AI Technical Summary
Vegetable fermentation broth has low utilization rate, and the market lacks special fermentation agents for different vegetables.
Microbial flora were extracted from the fermentation broth of different vegetables, and special fermentation was prepared, and applied to different vegetables through recombinant fermentation.
The utilization rate of vegetable fermentation broth is improved, and the quality of the prepared fermentation agent is better than that of commercially available products, and is suitable for different types of vegetables.
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Figure CN119799495A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of microorganisms, and in particular to a method for extracting microbial flora from vegetable fermentation liquid and applying recombinant fermentation. Background Art
[0002] Fermented vegetables are one of the most common vegetable processing methods. Under the action of microorganisms, they can give vegetables a unique flavor and extend the storage period of vegetables. There are many types of vegetables, including pickled radishes, pickled greens, pickled peppers, etc. In the process of fermented vegetable production, a large amount of vegetable fermentation liquid is produced, which contains a large amount of salt, organic acids, flavor substances and microorganisms. It is a veritable treasure trove of resources. However, with the rise of modern tank deep fermentation methods and the application of standardized direct-injection fermentation agents, the demand for vegetable fermentation liquid has dropped significantly, and there is an urgent need to find a method to efficiently utilize vegetable fermentation liquid.
[0003] Due to the different nutritional components of vegetable raw materials and the microorganisms attached to the vegetable interface, different vegetables have unique microbial flora, which together constitute a fermentation system. Related studies have also shown that vegetable fermentation liquid is rich in hundreds of microorganisms such as lactic acid bacteria and yeast, and there are more than ten dominant strains, all of which play an indispensable role. Although there are many composite bacterial vegetable fermentation agents on the market and they are widely used, the application scope and effect of the fermentation agents are limited because the composite bacterial strains generally do not exceed five. In addition, there are few special fermentation agent products for different vegetables on the market.
[0004] Based on this, the patent of the present invention extracts microbial flora from different vegetable fermentation broths, prepares special fermentation agents suitable for different vegetables, and recombines the fermentation for application in different vegetable fermentation to solve the problems of insufficient utilization of the above-mentioned vegetable fermentation broths and the temporary shortage of special fermentation agents for different vegetables. Summary of the invention
[0005] The present invention provides a method for extracting microbial flora from vegetable fermentation liquid and applying recombinant fermentation. Microbial flora are extracted from different vegetable fermentation liquids to prepare special fermentation agents for different vegetables. The recombinant fermentation is applied to fermentation of different vegetables to solve the problems of insufficient utilization of the above-mentioned vegetable fermentation liquids and temporary shortage of special fermentation agents for different vegetables.
[0006] In order to achieve the purpose of the present invention, the following technologies are proposed: On the one hand, a method for extracting microbial flora from vegetable fermentation broth is provided, comprising the steps of: Step 01, pre-treatment of the fermentation liquid, collecting the vegetable fermentation liquid, performing preliminary filtration and low-speed centrifugation on the obtained vegetable fermentation liquid to obtain a purified vegetable fermentation liquid; Step 02, enriching the microbial flora of the fermentation liquid, adjusting the pH of the purified vegetable fermentation liquid obtained in step 01, and then culturing and enriching it to obtain an enriched fermentation liquid; Step 03, collecting the microbial flora of the fermentation broth, centrifuging the enriched fermentation broth obtained in step 02, and collecting the microbial sludge of the fermentation broth; Step 04, preparation of fermentation liquid microbial flora agent, the fermentation liquid microbial sludge obtained in step 03 is mixed with a freeze-drying protective agent, and the pH value of the mixture is adjusted and then freeze-dried to obtain the fermentation liquid microbial flora agent.
[0007] Furthermore, the vegetable fermentation liquid can be selected from one of pickled radish fermentation liquid, pickled green vegetable fermentation liquid, pickled mustard fermentation liquid, pickled cowpea fermentation liquid, pickled cabbage fermentation liquid and pickled pepper fermentation liquid.
[0008] Further, in step 01, the vegetable fermentation liquid is initially filtered through a 200-300 mesh filter membrane; In step 01, a peristaltic pump is used at a flow rate of 3 to 5 L / min to allow the vegetable fermentation liquid that has undergone primary filtration to enter a tubular centrifuge. The tubular centrifuge performs centrifugal separation at a speed of 2000 r / min, collects the supernatant, and completes a low-speed centrifugation operation.
[0009] Furthermore, in step 02, the pH values of the various vegetable fermentation broths after re-filtration were adjusted to 6.0-6.5 by using 12.5% ammonia water.
[0010] Furthermore, in step 02, 10-30 g. / L glucose and 10-30 g / L yeast extract powder and 1-3 g / L osmotic pressure protective agent are added to the various vegetable fermentation broths after the pH is adjusted, and cultured at 37°C for 18 h~24 h.
[0011] Furthermore, in step 03, when the enriched fermentation broth obtained in step 02 is centrifugally filtered, a peristaltic pump is passed into a tubular centrifuge at a flow rate of 2 to 4 L / min, and centrifuged at a speed of 10,000 r / min in the tubular centrifuge. After centrifugation, the bacterial sludge in the inner cavity of the tubular centrifuge is collected.
[0012] Furthermore, in step 04, when preparing the fermentation broth microbial flora agent, the fermentation broth microbial sludge and the freeze-drying protective agent obtained in step 03 are taken, and the fermentation broth microbial sludge and the freeze-drying protective agent are mixed at a mass ratio of 1:0-2, and the pH of the mixture is adjusted to 6.5-7.0 by 12.5% ammonia water. The freeze-drying protective agent is 10%~15% trehalose, 5%~10% maltodextrin, and 1%~3% glycerol; When mixing, stir thoroughly in an ice-water bath for 15 to 30 min.
[0013] Furthermore, during freeze drying in step 04, the fermentation liquid microbial sludge mixed with the freeze-drying protective agent is poured into the freeze dryer tray, and the thickness of the material in the freeze dryer tray is 0.5-1.5 cm, and the freeze dryer tray is placed in an in-situ freeze dryer for freeze drying: When freeze-drying, pre-freeze at -20~-50℃ for 1~3h; Then carry out sublimation drying at -10~10℃ for 5~10h; Finally, analytical drying is carried out at 20-30°C for 8-15 h.
[0014] On the other hand, a method for recombinant fermentation of microbial flora in vegetable fermentation liquid is provided, comprising the steps of: Step 11, selecting vegetables to be fermented and processing the vegetables to be fermented; Step 12, adding salt water with a mass concentration of 5-12% to the vegetables to be fermented at a mass ratio of 1:1-1.5 between the vegetables to be fermented and the salt water; Step 13, adding 1-3% rock sugar, 1-2‰ star anise, 1-2‰ pepper, and 3-6‰ liquor to the vegetables to be fermented based on the total mass ratio of vegetables to water; Step 14, adding a fermentation liquid microbial flora agent at a total mass ratio of 1 to 3 parts per ten thousand; Step 15, ferment at 25-30°C for 10-20 days.
[0015] The advantages of the above technical solution are: Aiming at the problem of low utilization rate of vegetable fermentation liquid at present, the present invention prepares vegetable fermentation liquid microbial flora inoculant with high live bacteria count and good storage stability through the processes of vegetable fermentation liquid pretreatment, vegetable fermentation liquid microbial flora enrichment, vegetable fermentation liquid microbial flora collection and vegetable fermentation liquid microbial flora inoculant preparation.
[0016] In view of the problem of lack of special fermentation agents for different vegetables, the present invention prepares pickled radish fermentation liquid microbial flora agent, pickled green vegetable fermentation liquid microbial flora agent and pickled pepper fermentation liquid microbial flora agent through the above-mentioned vegetable fermentation liquid microbial flora agent preparation process, and then recombines and ferments the pickled radish, pickled green vegetables and pickled peppers respectively, and the prepared products are of high quality and are superior to commercially available fermentation agents. BRIEF DESCRIPTION OF THE DRAWINGS
[0017] In order to make the objectives, technical solutions and advantages of the present invention more clear, the present invention will be further described in detail below with reference to the accompanying drawings.
[0018] Figure 1 The effect of different ratios of fermentation sludge and protective agent on the bacterial agent is shown.
[0019] Figure 2 The response surface and contours of the interaction between glycerol and maltodextrin are shown.
[0020] Figure 3 The response surface and contour lines of the interaction between glycerol and trehalose are shown.
[0021] Figure 4 The response surface and contours of the interaction between trehalose and maltodextrin are shown. DETAILED DESCRIPTION
[0022] Comparative Example 1: Making pickled radish with commercially available fermentation agent Select fresh and undamaged radish, wash, drain and cut into long strips; add 5-8% salt water at a mass ratio of 1:1-1.5, and add 1-3% rock sugar, 1-2‰ star anise, 1-2‰ pepper and 3-6‰ liquor according to the total mass ratio of vegetable to water; then add commercial fermentation agent 1, commercial fermentation agent 2 and commercial fermentation agent 3 as pickled radish fermentation agents respectively at a ratio of 1-3 per ten thousand of the total mass ratio of vegetable to water, and ferment at 25-30℃ for 5-10 d.
[0023] Comparative Example 2: Making pickled vegetables with commercially available fermentation agents Select fresh and undamaged green vegetables, wash them and drain the water; add 5-8% salt water at a mass ratio of 1:1-1.5, and add 1-3% rock sugar, 1-2‰ star anise, 1-2‰ pepper, and 3-6‰ white wine according to the total mass ratio of vegetables to water; then add commercial fermentation agent 1, commercial fermentation agent 2, and commercial fermentation agent 3 as pickled green vegetable fermentation agents at a ratio of 1-3 per ten thousand of the total mass ratio of vegetables to water, and ferment at 25-30℃ for 5-10 days.
[0024] Comparative Example 3: Making pickled peppers with commercially available fermentation agents Select fresh and undamaged pickled peppers, wash and drain, add 8-12% brine at a mass ratio of 1:1-1.5, and add 1-3% rock sugar, 1-2‰ star anise, 1-2‰ pepper, and 3-6‰ white wine according to the total mass ratio of vegetables to water; then add commercially available fermentation agent 1, commercially available fermentation agent 2, and commercially available fermentation agent 3 as pickled pepper fermentation agents according to the ratio of 1-3 per ten thousand of the total mass ratio of vegetables to water, and ferment at 30-35℃ for 15-25 days.
[0025] Example 1 A method for extracting microbial flora from vegetable fermentation liquid comprises the steps of: Step 01, pre-treatment of the fermentation liquid, respectively collecting the pickled radish fermentation liquid, pickled green vegetable fermentation liquid and pickled pepper fermentation liquid, respectively performing primary filtration and low-speed centrifugation on the obtained vegetable fermentation liquid to obtain purified vegetable fermentation liquid.
[0026] During the initial filtration, the vegetable fermentation liquid is filtered through a 200-300 mesh filter membrane to remove large particles of insoluble impurities.
[0027] During refiltration, a peristaltic pump is used at a flow rate of 3 to 5 L / min to allow the vegetable fermentation liquid that has passed the initial filtration to enter the tubular centrifuge. The tubular centrifuge performs centrifugal separation at a speed of 2000 r / min and collects the supernatant. Low-speed centrifugation can remove small particles of insoluble impurities again.
[0028] Step 02, enrichment of the microbial flora of the fermentation liquid, adjusting the pH of the vegetable fermentation liquid obtained in step 01, culturing and enriching it to obtain an enriched fermentation liquid.
[0029] Among them, the pH value of each vegetable fermentation liquid after re-filtration was adjusted to 6.0~6.5 by 12.5% ammonia water.
[0030] During the culture, 10-30 g. / L glucose, 10-30 g / L yeast extract powder and 1-3 g / L proline (osmotic pressure protectant) were added to each vegetable fermentation broth after pH adjustment, and cultured at 37°C for 18 h~24 h.
[0031] Table 1: Orthogonal test design and results of carbon source, nitrogen source and osmotic pressure protectant addition As shown in Table 1, the addition amounts of carbon source, nitrogen source and osmotic pressure protectant were optimized through three-factor three-level orthogonal test. Finally, 30 g / L glucose, 30 g / L yeast extract and 2 g / L proline were required for the enrichment of microbial flora in the fermentation broth.
[0032] Step 03, collection of fermentation broth microbial flora, centrifugation of each enriched fermentation broth obtained in step 02, and collection of fermentation broth microbial sludge.
[0033] When each fermentation broth after enrichment is filtered through centrifugation, the peristaltic pump is passed into a tubular centrifuge at a flow rate of 2-4 L / min, and centrifuged at a speed of 10,000 r / min in the tubular centrifuge. After centrifugation, the bacterial sludge in the inner cavity of the tubular centrifuge is collected.
[0034] Step 04, preparation of fermentation broth microbial flora agent, respectively mixing each fermentation broth microbial sludge obtained in step 03 with a freeze-drying protective agent, stirring the mixture in an ice water bath for 15 to 30 minutes, adjusting the pH value of the mixture and then freeze-drying to obtain a fermentation broth microbial flora agent.
[0035] When preparing the fermentation liquid microbial flora agent, the fermentation liquid microbial sludge and freeze-drying protective agent obtained in step 03 are taken, and the mass ratio of each fermentation liquid microbial sludge to freeze-drying protective agent is 1:0-2, and the pH of the mixture is adjusted to 6.5-7.0 by adjusting the mixture with 12.5% ammonia water. Among them, the freeze-drying protective agent is 10%-15% trehalose, 5%-10% maltodextrin, and 1%-3% glycerol.
[0036] During freeze drying, the fermentation liquid microbial sludge mixed with a freeze-drying protective agent is poured into the freeze dryer tray, and the thickness of the material in the freeze dryer tray is 0.5-1.5 cm. The freeze dryer tray is placed in an in-situ freeze dryer for freeze drying.
[0037] When freeze-drying, pre-freeze at -20~-50℃ for 1~3 h.
[0038] Then carry out sublimation drying at -10~10℃ for 5~10 h.
[0039] Finally, analytical drying is carried out at 20-30°C for 8-15 h.
[0040] Table 1: Orthogonal test design and results of carbon source, nitrogen source and osmotic pressure protectant addition Table 2: Three-factor three-level response surface design and results of lyophilized protective agents Serial number A: Trehalose / (%) B: Maltodextrin / (%) C: Glycerol / (%) Number of viable bacteria / (lg CFU / g) 1 10 7.5 3 7.05 2 12.5 7.5 2 8.93 3 12.5 7.5 2 9.30 4 12.5 7.5 2 9.08 5 15 7.5 1 8.56 6 10 7.5 1 8.04 7 12.5 5 3 7.37 8 15 5 2 6.03 9 12.5 7.5 2 9.00 10 12.5 10 1 10.59 11 10 5 2 5.25 12 12.5 7.5 2 9.37 13 12.5 10 3 9.67 14 12.5 5 1 6.7 15 10 10 2 8.78 16 15 7.5 3 9.60 17 15 10 2 10.94 Table 3: Analysis of variance of response surface test results factor sum of squares Degrees of Freedom Mean Square value value Significance Model 37.91 9 4.21 37.37 <0.0001 ** A-Trehalose 4.51 1 4.51 40.02 0.0004 ** B-Maltodextrin 26.81 1 26.81 237.87 <0.0001 ** C-Glycerol 4.841E-003 1 4.841E-003 0.043 0.8417 AB 0.48 1 0.48 4.27 0.0775 AC 1.03 1 1.03 9.13 0.0193 * BC 0.63 1 0.63 5.61 0.0497 A^2 2.9 1 2.9 25.7 0.0014 ** B^2 1.31 1 1.31 11.59 0.0114 * C^2 2.306E-005 1 2.306E-005 2.045E-004 0.989 error 0.79 7 0.11 Lack of Fit 0.64 3 0.21 5.76 0.062 Not significant Pure Error 0.15 4 0.037 sum 38.7 16 According to GB4789.35-2016 (National Food Safety Standard - Food Microbiology Test - Lactic Acid Bacteria Test), the number of live bacteria in different samples was measured, and the survival rate was calculated based on the total number of live bacteria before and after freeze-drying. Figure 1 It can be seen that with the increase of the amount of protective agent added, the number of live bacteria in the powder first increased and then decreased, while the survival rate of the powder gradually increased. Considering the number of live bacteria and the survival rate, the best ratio of the dry weight of the sludge to the protective agent was 1:1, and the number of live bacteria reached 6.63×10 10 CFU / g, and the survival rate was 76.4%, which were increased by 86.24% and 131.3% respectively compared with the samples without adding protective agents.
[0041] Through regression analysis and fitting of the data in Table 2, see Table 3 and Figure 2 Thus, a regression equation fitting the experimental factors was obtained: Y=9.14+0.75A+1.83B-0.025C+0.35AB+0.51AC-0.40BC-0.83A 2 -0.56B 2 -0.0002C2 The experimental model P The value was <0.01, which indicates that the high and low levels of each factor have a very significant effect on the viable count of the response value. The P value of the lack of fit term was 0.062>0.05, which was not significant, indicating that the model had a good fit and could be effectively analyzed and predicted. The coefficient of the quadratic term was negative, indicating that the model had a maximum value. By solving the maximum value of the regression equation, the optimal addition amount of each factor was 13.4% trehalose, 10.0% maltodextrin and 1.0% glycerol (the remainder was sterile distilled water). Under this condition, the viable count of the bacterial powder was 10.95 lg CFU / g, the survival rate was 86.92%, and the viable count was still greater than 1×10 after 365 days of storage at 4℃. 10 CFU / g, with high viable count and storage stability.
[0042] According to the above steps, one of the pickled radish fermentation liquid microbial flora agent, pickled green vegetable fermentation liquid microbial flora agent, pickled pepper fermentation liquid microbial flora agent, pickled mustard fermentation liquid microbial flora agent, pickled cabbage fermentation liquid microbial flora agent or pickled cowpea fermentation liquid microbial flora agent is prepared respectively for easy reorganization application.
[0043] Example 2: Making pickled radish with fermentation agent Select fresh and undamaged radish, wash, drain and cut into long strips; add 5-8% salt water at a mass ratio of 1:1-1.5, and add 1-3% crystal sugar, 1-2‰ star anise, 1-2‰ pepper and 3-6‰ liquor at a total mass ratio; then add the pickled radish fermentation liquid microbial flora inoculant prepared in Example 1 as a pickled radish fermentation agent at a total mass ratio of 1-3 per ten thousand, and ferment at 25-30°C for 5-10 days.
[0044] Example 3: Making pickled vegetables with a starter Select fresh and undamaged green vegetables, wash them, and drain them; add 5-8% salt water at a mass ratio of 1:1-1.5, and add 1-3% crystal sugar, 1-2‰ star anise, 1-2‰ pepper, and 3-6‰ liquor at a total mass ratio; then add the pickled green vegetable fermentation liquid microbial flora inoculant prepared in Example 1 as the pickled green vegetable fermentation agent at a total mass ratio of 1-3 per ten thousand, and ferment at 25-30°C for 5-10 days.
[0045] Example 4: Making pickled peppers with a starter Select fresh and undamaged pickled peppers, wash and drain, add 8-12% salt water at a mass ratio of 1:1-1.5, and add 1-3% crystal sugar, 1-2‰ star anise, 1-2‰ pepper, and 3-6‰ liquor according to the total mass ratio; then add the pickled pepper fermentation liquid microbial flora inoculant prepared in Example 1 as the pickled pepper fermentation agent at a ratio of 1-3 per ten thousand of the total mass ratio, and ferment at 30-35°C for 15-25 d.
[0046] Table 4: Sensory evaluation criteria Table 5: Sensory evaluation results of each sample Sensory evaluation can intuitively compare the differences in product color, shape, smell and taste. The pickled radish, pickled greens and pickled peppers prepared in Examples 2, 3 and 4 were compared with the products prepared in Comparative Examples 1, 2 and 3. Ten tasters (5 males and 5 females, aged between 25 and 45) with a food professional background and professional sensory training were selected. The scores of all the judges were counted from the four aspects of color, smell, taste and texture, and the scores were averaged. The specific scoring criteria are shown in Table 4, and the scoring results are shown in Table 5.
[0047] As shown in Table 5, the pickled radish, pickled greens and pickled pepper fermentation broth microbial flora inoculants prepared in Example 1 were used to inoculate and ferment pickled radish, pickled greens and pickled peppers, respectively. The products were superior to the commercially available fermentation agents through sensory evaluation. Microbial flora were extracted from different fermentation broths, prepared into inoculants, and then recombined for fermentation. Because the flora structure was more suitable for raw material fermentation, the product quality was significantly improved.
[0048] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.
Claims
1. A method for extracting microbial flora from vegetable fermentation broth, characterized in that: Includes steps: Step 01, pre-treatment of the fermentation liquid, collecting the vegetable fermentation liquid, and performing preliminary filtration and centrifugation on the obtained vegetable fermentation liquid to obtain a purified vegetable fermentation liquid; Step 02, enriching the microbial flora of the fermentation liquid, adjusting the pH of the purified vegetable fermentation liquid obtained in step 01, and then culturing and enriching it to obtain an enriched fermentation liquid; Step 03, collecting the microbial flora of the fermentation broth, centrifuging the enriched fermentation broth obtained in step 02, and collecting the microbial sludge of the fermentation broth; Step 04, preparation of fermentation liquid microbial flora agent, the fermentation liquid microbial sludge obtained in step 03 is mixed with a freeze-drying protective agent, and the pH value of the mixture is adjusted and then freeze-dried to obtain the fermentation liquid microbial flora agent.
2. The method for extracting microorganisms from vegetable fermentation liquid according to claim 1, characterized in that: The vegetable fermented liquid is one of pickled radish fermented liquid, pickled green vegetable fermented liquid, pickled mustard fermented liquid, pickled cowpea fermented liquid, pickled cabbage and pickled pepper fermented liquid.
3. The method for extracting microorganisms from vegetable fermentation liquid according to claim 1, characterized in that: In step 01, the vegetable fermentation liquid is initially filtered through a 200-300 mesh filter membrane; In step 01, a peristaltic pump is used at a flow rate of 3 to 5 L / min to allow the vegetable fermentation liquid that has undergone primary filtration to enter a tubular centrifuge. The tubular centrifuge performs centrifugal separation at a speed of 2000 r / min, and collects the supernatant to complete the centrifugation operation.
4. The method for extracting microorganisms from vegetable fermentation liquid according to claim 1, characterized in that: In step 02, the pH values of the re-filtered vegetable fermentation broths are adjusted to 6.0-6.5 using 12.5% ammonia water.
5. The method for extracting microorganisms from vegetable fermentation liquid according to claim 1, characterized in that: In step 02, 10-30 g. / L glucose, 10-30 g / L yeast extract powder and 1-3 g / L proline are added to the various vegetable fermentation broths after pH adjustment, and cultured at 37°C for 18 h~24 h.
6. The method for extracting microorganisms from vegetable fermentation liquid according to claim 1, characterized in that: In step 03, when the enriched fermentation broth obtained in step 02 is centrifugally filtered, a peristaltic pump is used to pass it into a tubular centrifuge at a flow rate of 2 to 4 L / min, and centrifuged at a speed of 10,000 r / min in the tubular centrifuge. After centrifugation, the bacterial sludge in the inner cavity of the tubular centrifuge is collected.
7. The method for extracting microorganisms from vegetable fermentation liquid according to claim 1, characterized in that: In step 04, when preparing the fermentation liquid microbial flora agent, the fermentation liquid microbial sludge and the freeze-drying protective agent obtained in step 03 are taken, and the fermentation liquid microbial sludge and the freeze-drying protective agent are mixed at a mass ratio of 1:0-2, and the pH of the mixture is adjusted to 6.5-7.0 by 12.5% ammonia water. The freeze-drying protective agent is 10%~15% trehalose, 5%~10% maltodextrin, and 1%~3% glycerol; When mixing, stir thoroughly in an ice-water bath for 15 to 30 min.
8. The method for extracting microorganisms from vegetable fermentation liquid according to claim 1, characterized in that: During freeze drying in step 04, the fermentation liquid microbial sludge mixed with the freeze-drying protective agent is poured into the freeze dryer tray, and the thickness of the material in the freeze dryer tray is 0.5-1.5 cm, and the freeze dryer tray is placed in the in-situ freeze dryer for freeze drying: When freeze-drying, pre-freeze at -20~-50℃ for 1~3h; Then carry out sublimation drying at -10~10℃ for 5~10 h; Finally, analytical drying is carried out at 20-30°C for 8-15 h.
9. A method for the application of recombinant fermentation of microbial flora in vegetable fermentation broth, characterized in that: Includes steps: Step 11, selecting vegetables to be fermented and processing the vegetables to be fermented; Step 12, adding salt water with a mass concentration of 5-12% to the vegetables to be fermented at a mass ratio of 1:1-1.5 between the vegetables to be fermented and the salt water; Step 13, adding 1-3% rock sugar, 1-2‰ star anise, 1-2‰ pepper, and 3-6‰ liquor to the vegetables to be fermented according to the total mass ratio; Step 14, adding a fermentation liquid microbial flora agent prepared by the method for extracting microbial flora from vegetable fermentation liquid according to any one of claims 1 to 8 at a ratio of 1 to 3 per 10,000 of the total mass ratio of vegetable to water; Step 15, ferment at 25-30°C for 10-20 days.