Processing technology of polygonum multiflorum and prepared polygonum multiflorum
Through the infiltration and steaming of black bean juice and He Shouwu slices, combined with the steps of simplicity and closed fermentation, successfully screening and cultivating Xiev's shades mold, solving the problem of cumbersome and limited microbial applications in the existing technologies, and realized that the microbial application is limited, realizing the realization, realizing the realization of the realization, realization The high content and safety of the production of Xieva Gem in Hewei Shouwu is the safety of Xiewa.
Patent Information
- Application Number
- CN202411563714.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-05
- Publication Date
- 2025-01-24
AI Technical Summary
The existing process of processing Polygonum multiflorum is complicated, and it is difficult to fully utilize the efficacy of Polygonum multiflorum, and there are problems of risk of poisoning and quality failure. The application of microbial fermentation method in traditional Chinese medicine materials is limited, making it difficult to screen and cultivate specific microorganisms in directional screening and cultivation.
Using black bean juice and He Shouwu slices to hybridize and steam, then simmer and dry it, and finally ferment under closed conditions. Growth and activity.
The directional screening and cultivation of Aspergillus sievai on Polygonum multiflorum has been realized, the quantity and purity of Aspergillus siewani in the preparation of Polygonum multiflorum have been improved, biological transformation has been promoted, the range of microbial species has been broadened, and the safety of the product has been ensured.
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Figure CN119345266A_ABST
Abstract
Description
Technical Field
[0001] The present application belongs to the technical field of traditional Chinese medicine processing and biotransformation, and specifically relates to a processing technology of Polygonum multiflorum and the prepared Polygonum multiflorum. Background Art
[0002] Polygonum multiflorum is the dried root of Polygonum multiflorum Thumb, a plant of the Polygonaceae family, and is one of the representative varieties of raw and cooked Chinese medicine. For the processing of Polygonum multiflorum, the traditional method of nine steaming and nine drying is widely used, but the process is cumbersome and not suitable for modern industrial production; modern methods mostly use steaming and black bean processing, but the above two methods are still difficult to fully exert the efficacy of processed Polygonum multiflorum, and there have even been reports of poisoning caused by processed Polygonum multiflorum. It is not only time-consuming and labor-intensive, but also has no quality assurance and causes waste of raw materials.
[0003] In recent years, microbial fermentation has attracted more and more attention. During the processing, the enzymes produced by microorganisms can specifically transform or degrade the toxic components in traditional Chinese medicine, realize the transformation of microorganisms, without affecting the content of effective ingredients, and even promote the increase or activity enhancement of certain components of traditional Chinese medicine. This processing method is more in line with the processing purpose of "increasing efficacy and reducing toxicity" in traditional Chinese medicine processing. Although microbial transformation is widely used in the field of traditional Chinese medicine, the general microbial fermentation method uses natural bacteria in the environment, such as bacteria, molds, yeasts and other strains for fermentation. The purity of the strains is not high and is greatly affected by the environment. The microbial strains are messy and not targeted and selective, resulting in microorganisms that are not dominant in nature and have strict growth conditions. It is difficult to grow on the medicine. Therefore, the range of microbial species is limited, and the potential efficacy of microorganisms in the medicine is difficult to be effectively exerted. In addition, some strains that fall into the natural fermentation process are also toxic, and the quality and safety of fermented products cannot be guaranteed. How to obtain a fixed fermentation flora on traditional Chinese medicine materials to improve and ensure the production quality of traditional Chinese medicine is still an unknown problem.
[0004] Aspergillus chevalieri is a dominant fungus in the natural fermentation process of tea, commonly known as the "golden flower fungus". It is often used in the fermentation of tea and plays an indispensable role in improving the quality of tea. Under the action of Aspergillus chevalieri, tea can produce a unique fungus flower fragrance and mellow taste. Therefore, the number of Aspergillus chevalieri has gradually become a criterion for evaluating the quality of tea. New research has found that the production of Aspergillus chevalieri can also give fermented tea certain health functions, such as reducing fat and weight, promoting digestion, and anti-tumor (references [Cao Chang. Construction and functional analysis of Aspergillus chevalieri intermorph △ ku70 strain [D]. Hubei: Huazhong Agricultural University, 2013.], [Liu Rong, Tan Yumei, Liu Yongxiang, et al. Functional analysis of edsc gene of Aspergillus chevalieri intermorph [J]. Journal of Mycology, 2015, 32(2): 227-234.]). As mentioned above, the current application of Aspergillus Shewani is mainly concentrated in the fermentation of tea, and its research in traditional Chinese medicinal materials is relatively unclear. At present, the application of Aspergillus Shewani in the processing of Polygonum multiflorum has not been found. Summary of the invention
[0005] The purpose of the present application is to overcome the shortcomings of the prior art, provide a processing process for Polygonum multiflorum, and prepare a processed Polygonum multiflorum product containing Aspergillus chevalieri, which opens up a new path for the diversified application of Aspergillus chevalieri and a new processing method for Polygonum multiflorum.
[0006] In order to achieve the above-mentioned invention object, the present application provides a method for processing Polygonum multiflorum, characterized in that it comprises the following steps:
[0007] (1) mixing the Polygonum multiflorum slices and the black bean juice, infiltrating them, separating the solid from the liquid, and collecting the infiltrated black bean juice and the infiltrated Polygonum multiflorum slices;
[0008] (2) steaming the soaked Polygonum multiflorum slices obtained in step (1) to obtain steamed Polygonum multiflorum slices; the steaming temperature is 85 to 100° C.; the steaming time is 22 to 26 hours;
[0009] (3) stewing the steamed Polygonum multiflorum slices obtained in step (2), mixing the stewed Polygonum multiflorum slices with the soaked black bean juice obtained in step (1), and air-drying the mixed material to obtain dried Polygonum multiflorum slices;
[0010] (4) fermenting the dried Polygonum multiflorum slices obtained in step (3) under closed conditions to obtain processed Polygonum multiflorum.
[0011] In addition, this application also provides the following subsidiary technical solutions:
[0012] Preferably, the method for preparing black bean juice in step (1) comprises the following steps:
[0013] 1) mixing black beans with water for a first cooking, and separating a first liquid component and bean dregs;
[0014] 2) mixing the okara in step 1) with water for a second cooking, and collecting a second liquid component;
[0015] 3) combining the first liquid component and the second liquid component to obtain black bean juice.
[0016] In the step 1), the mass volume ratio of black beans to water is 1 kg: (2.8-3.2) L; the first cooking time is 3.5-4.5 h.
[0017] The mass volume ratio of the okara to water in the step 2) is 1 kg: (2.8-3.2) L; the second cooking time is 2.5-3.5 h.
[0018] Preferably, the mass ratio of the Polygonum multiflorum slices to the black bean juice in the step (1) is 10:(2.1-2.9).
[0019] Preferably, the infiltration time in step (1) is 22 to 26 hours.
[0020] Preferably, the stewing time in step (3) is 1.5 to 2.5 hours.
[0021] Preferably, the fermentation time in step (4) is 6 to 8 days.
[0022] Preferably, the fermentation temperature in step (4) is 20-40°C.
[0023] The present application also provides a processed Polygonum multiflorum, which is prepared by the above-mentioned processing method. The effective viable count of Aspergillus chevalieri in the processed Polygonum multiflorum is (250,000) to (310,000) CFU / g.
[0024] The beneficial effects of this application are:
[0025] The present application discloses a processing process of Polygonum multiflorum, which uses black bean juice to steam and ferment Polygonum multiflorum, increases the quantity and purity of Aspergillus chevalieri in black bean juice, and realizes the directional screening and cultivation of Aspergillus chevalieri on Polygonum multiflorum, so that microorganisms with strict growth conditions such as Aspergillus chevalieri can effectively grow on Polygonum multiflorum, and finally obtain a processed Polygonum multiflorum product containing a higher content of Aspergillus chevalieri. The processing method provided by the present application is conducive to promoting the biotransformation in the processing process of Polygonum multiflorum, and broadens the range of microbial species accessed by Polygonum multiflorum, which lays a foundation for the research on the potential efficacy of Aspergillus chevalieri in traditional Chinese medicine, especially in Polygonum multiflorum medicine. In addition, the processed Polygonum multiflorum product prepared by the present application does not contain toxic and harmful microorganisms and their products (such as Aspergillus flavus, Escherichia coli, Salmonella and aflatoxin), which can ensure the safety of the processed Polygonum multiflorum product fermented by He Shouwu. BRIEF DESCRIPTION OF THE DRAWINGS
[0026] In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.
[0027] Figure 1 The counting results of sample A TAMC (left) and TAYC (right) are shown;
[0028] Figure 2 This is the counting result diagram of TAMC (left) and TAYC (right) of Polygonum multiflorum No. 1 sample;
[0029] Figure 3 This is the counting result diagram of TAMC (left) and TAYC (right) of Polygonum multiflorum No. 1 sample;
[0030] Figure 4 The results of TAMC (left) and TAYC (right) counting of Radix Polygoni Multiflori A are shown;
[0031] Figure 5 The results of TAMC (left) and TAYC (right) counting of Radix Polygoni Multiflori B are shown;
[0032] Figure 6 The colony morphology of Aspergillus shevarii in sample 1 of Polygonum multiflorum preparation No. 1;
[0033] Figure 7 For the colony morphology of Aspergillus shevarii in sample 2 of Polygonum multiflorum preparation No. 1;
[0034] Figure 8This is the colony morphology of Aspergillus shevarii in sample 3 of Polygonum multiflorum No. 1. DETAILED DESCRIPTION
[0035] The present application provides a method for processing Polygonum multiflorum, characterized in that it comprises the following steps:
[0036] (1) mixing the Polygonum multiflorum slices with black bean juice, infiltrating them, separating the solid from the liquid, and collecting the infiltrated black bean juice and the infiltrated Polygonum multiflorum slices;
[0037] (2) steaming the soaked Polygonum multiflorum slices obtained in step (1) to obtain steamed Polygonum multiflorum slices; the steaming temperature is 85 to 100° C.; the steaming time is 22 to 26 hours;
[0038] (3) stewing the steamed Polygonum multiflorum slices obtained in step (2), mixing the stewed Polygonum multiflorum slices with the soaked black bean juice obtained in step (1), and air-drying the mixed material to obtain dried Polygonum multiflorum slices;
[0039] (4) fermenting the dried Polygonum multiflorum slices obtained in step (3) under closed conditions to obtain processed Polygonum multiflorum.
[0040] In the present application, the thickness of the slices in the step (1) is preferably 0.8 to 1.2 cm, more preferably 1 cm; the drying in the step (1) or the step (2) is preferably carried out at room temperature of 20 to 25°C, more preferably 25°C. The present application does not specifically limit the source of the raw Polygonum multiflorum, and the raw Polygonum multiflorum conventionally prepared by technicians in this field can be used. In the present application, slicing can increase the contact area between Polygonum multiflorum and black bean juice, making it easier for Aspergillus Shewanella to be inoculated onto the Polygonum multiflorum medicinal material, and while ensuring the steaming effect, it can ensure that the quality of Polygonum multiflorum is stable and controllable.
[0041] In the present application, the preparation of black bean juice in step (1) of the processing method of Polygonum multiflorum preferably comprises the following steps:
[0042] 1) mixing black beans with water for a first cooking, and separating a first liquid component and bean dregs;
[0043] 2) mixing the okara in step 1) with water for a second cooking, and collecting a second liquid component;
[0044] 3) combining the first liquid component and the second liquid component to obtain black bean juice.
[0045] In the present application, the mass volume ratio of black beans to water in the step 1) is more preferably 1kg:(2.8-3.2)L, and more preferably 1kg:3L; the first cooking time is more preferably 3.5-4.5h, and more preferably 4h. In the present application, the mass volume ratio of dregs to water in the step 2) is more preferably 1kg:(2.8-3.2)L, and more preferably 1kg:3L; the second cooking time is more preferably 2.5-3.5h, and more preferably 3h. In the present application, the separation in the step 1) is more preferably carried out by filtration. The collection of the second liquid component in the step 2) is more preferably carried out by filtration. The filtration is further preferably carried out by filtering with a filter screen, and further preferably by filtering with a 100-mesh filter screen. The present application does not specifically limit the source of the black beans, and the black beans commonly used in the conventional black bean juice preparation by those skilled in the art can be used. In the present application, black bean juice prepared by a specific preparation method is used as a processing auxiliary material, which can not only reduce the toxicity of the raw Polygonum multiflorum, but also enhance its original efficacy. After two boilings with water, the obtained black bean juice is of high quality, and the active ingredients in the black beans can be fully boiled into the black bean juice, which can improve the processing effect of the black bean juice on the Polygonum multiflorum, and can also provide a rich material basis for the subsequent growth of Aspergillus Shewanella. While improving the processing quality of Polygonum multiflorum, it can also further increase the number of Aspergillus Shewanella in the subsequent fermentation process of the black bean juice.
[0046] In the present application, the mass ratio of the Polygonum multiflorum slices to the black bean juice in step (1) of the Polygonum multiflorum processing method is preferably 10:(2.3-2.9), more preferably 10:2.5. In the Polygonum multiflorum processing process of the present application, adding a specific proportion of black bean juice as a processing auxiliary material can improve the processing quality of Polygonum multiflorum, and can also provide a rich material basis for the subsequent growth of Aspergillus Shewanella, thereby further increasing the number of Aspergillus Shewanella in the subsequent fermentation process of black bean juice.
[0047] In the present application, the infiltration in step (1) is preferably carried out in a non-iron container. In the present application, the selection of a non-iron container can prevent some active ingredients in the processing of Polygonum multiflorum from reacting with iron, resulting in loss of efficacy, and prevent the growth of Aspergillus shevarii from being affected.
[0048] In the present application, the soaking time in step (1) is preferably 22 to 26 hours, more preferably 24 hours. In the present application, the soaking can make the Polygonum multiflorum fully absorb the black bean juice, improve the processing quality of the Polygonum multiflorum, and the rich material basis can further promote the growth of Aspergillus shevarii in the subsequent fermentation process of the black bean juice.
[0049] In the present application, the steaming temperature in step (2) is more preferably 100°C; the steaming time is more preferably 24h. In the present application, the use of steaming to heat Polygonum multiflorum has the effects of enhancing the therapeutic effect, alleviating the medicinal properties, facilitating storage, and facilitating softening. In addition, the specific steaming conditions can kill pests and harmful bacteria in the raw Polygonum multiflorum by circulating steam, but cannot achieve a complete sterilization effect on the spores of Aspergillus serrata. In the subsequent fermentation process of Polygonum multiflorum with black bean juice, Aspergillus serrata can be produced in large quantities and can become the dominant bacteria in natural fermentation.
[0050] In the present application, the stewing time in step (3) is preferably 1.5 to 2.5 hours, more preferably 2 hours. In the present application, the stewing and the step of drying after adding the soaked black bean juice in step (1) can make the black bean juice fully penetrate into the internal tissue of the Polygonum multiflorum medicinal material to achieve internal and external consistency, thereby improving the processing quality of Polygonum multiflorum and ensuring the nutritional conditions of Aspergillus shevarii in the subsequent fermentation process of the black bean juice.
[0051] In the present application, the time of the closed fermentation in step (4) is preferably 6 to 8 days, more preferably 7 days. In the present application, the temperature of the closed fermentation in step (4) is preferably 20 to 40°C, more preferably 25°C. In the present application, the use of a specific closed fermentation can make the Aspergillus Shevarii produced in large quantities during the fermentation process after steaming black beans, and avoid contamination by miscellaneous bacteria, further increase the number and purity of Aspergillus Shevarii in the subsequent fermentation process of black bean juice, and facilitate the growth of Aspergillus Shevarii on Polygonum multiflorum.
[0052] The present application also provides a processed Polygonum multiflorum prepared by the above-mentioned processing method, wherein the processed Polygonum multiflorum contains Aspergillus chevalieri; the effective viable count of Aspergillus chevalieri is (250000) to (310000) CFU / g, more preferably (310000) CFU / g. g .
[0053] In order to further illustrate the present application, the processing technology of Polygonum multiflorum and the prepared Polygonum multiflorum provided by the present application are described in detail below in combination with the drawings and examples, but they should not be understood as limiting the scope of protection of the present application.
[0054] Example 1
[0055] 1. Preparation of Black Bean Juice
[0056] Take 1kg of black beans, add 3L of water, cook for 4 hours, and filter the dregs with a 100-mesh filter to get the juice.
[0057] 1.5kg, add 3 times the volume of water of the bean dregs to the obtained bean dregs, boil again for 3h, filter the bean dregs with a 100-mesh filter again to obtain 1.0kg of boiled juice, combine the two boiled juices to obtain 2.5kg of black bean juice.
[0058] 2. Preparation of Polygonum multiflorum No. 1
[0059] Take 10 kg of raw Polygonum multiflorum and slice it into 1 cm thick slices, wash it with distilled water, dry it at 25°C, sort it by size, add 2.5 kg of black bean juice and mix it well, put it in a suitable non-iron container, soak it for 24 hours, let the black bean juice fully absorb into the slices, steam it at 100°C for 24 hours, then simmer it for 2 hours until the slices appear dark brown, mix it with the remaining black bean juice, and dry it at 25°C. Put the dried Polygonum multiflorum slices in a sealed bag, ferment them at 25°C for 7 days, and name it as "Processed Polygonum multiflorum Han No. 1" sample 1.
[0060] Example 2
[0061] 1. Preparation of Black Bean Juice
[0062] Take 1kg of black beans, add 2.8L of water, cook for 3.5h, filter the dregs with a 100-mesh filter to get 1.3kg of juice, add 2.8 times the volume of water of the dregs to the dregs, cook again for 2.5h, filter the dregs with a 100-mesh filter again to get 0.8kg of juice, combine the two juices to get 2.1kg of black bean juice.
[0063] 2. Preparation of Polygonum multiflorum No. 1
[0064] Take 10kg of raw Polygonum multiflorum and slice it. The slices are 0.8cm thick, washed with distilled water, dried at 20℃, graded by size, added with 2.1kg of black bean juice and mixed well, placed in a suitable non-iron container, soaked for 22h, so that the black bean juice is fully absorbed into the slices, steamed at 85℃ for 22h, and then simmered for 1.5h until the slices appear dark brown, mixed with the remaining black bean juice, and dried at 20℃. The dried Polygonum multiflorum slices are placed in a sealed bag and fermented at 20℃ for 8d, named "Processed Polygonum multiflorum I" sample 2.
[0065] Example 3
[0066] Take 1kg of black beans, add 3.2L of water, cook for 4.5h, filter the dregs with a 100-mesh filter to get 1.7kg of juice, add 3.2 times the volume of water of the dregs to the dregs, cook again for 3.5h, filter the dregs with a 100-mesh filter again to get 1.2kg of juice, combine the two juices to get 2.9kg of black bean juice.
[0067] 2. Preparation of Polygonum multiflorum No. 1
[0068] Take 10 kg of raw Polygonum multiflorum and slice it. The slices are 1.2 cm thick, washed with distilled water, dried at 23°C, graded by size, added with 2.9 kg of black bean juice, mixed well, placed in a suitable non-iron container, soaked for 26 hours, so that the black bean juice is fully absorbed into the slices, steam heated at 95°C for 26 hours, and then simmered for 2.5 hours until the slices appear dark brown, mixed with the remaining black bean juice, and dried at 23°C. The dried Polygonum multiflorum slices are placed in a sealed bag and fermented at 40°C for 6 days, named "Processed Polygonum multiflorum I" sample 3.
[0069] Comparative Example 1
[0070] The difference between Comparative Example 1 and Example 1 is that no black bean juice is added and no natural fermentation is performed:
[0071] 10 kg of raw Polygonum multiflorum was sliced into 1 cm thick slices, washed with distilled water, air-dried at 25° C., divided into different sizes, steamed at 100° C. for 24 h, and air-dried at 25° C. The dried Polygonum multiflorum slices were set aside and named as processed Polygonum multiflorum A.
[0072] Comparative Example 2
[0073] The difference between Comparative Example 1 and Example 1 is that natural fermentation is not performed:
[0074] 1. Preparation of Black Bean Juice
[0075] Take 1kg of black beans, add 3L of water, cook for 4 hours, and filter the dregs with a 100-mesh filter to get the juice.
[0076] 1.5kg, add 3 times the volume of water of the bean dregs to the obtained bean dregs, boil again for 3h, filter the bean dregs with a 100-mesh filter again to obtain 1.0kg of boiled juice, combine the two boiled juices to obtain 2.5kg of black bean juice.
[0077] 2. Preparation of Polygonum multiflorum No. 1
[0078] Take 10 kg of raw Polygonum multiflorum and slice it into 1 cm thick slices, wash it with distilled water, dry it at 25°C, sort it by size, add 2.5 kg of black bean juice and mix well, put it in a suitable non-iron container, soak it for 24 hours to make the black bean juice fully absorbed into the slices, steam it at 100°C for 24 hours, then simmer it for 2 hours until the slices turn dark brown, mix it with the remaining black bean juice, dry it at 25°C, slice the dried Polygonum multiflorum for later use, and name it processed Polygonum multiflorum B.
[0079] Comparative Example 3
[0080] The difference between Comparative Example 3 and Example 1 is only the difference in steaming conditions:
[0081] 1. Preparation of Black Bean Juice
[0082] Take 1kg of black beans, add 3L of water, cook for 4 hours, and filter the dregs with a 100-mesh filter to get the juice.
[0083] 1.5kg, add 3 times the volume of water of the bean dregs to the obtained bean dregs, boil again for 3h, filter the bean dregs with a 100-mesh filter again to obtain 1.0kg of boiled juice, combine the two boiled juices to obtain 2.5kg of black bean juice.
[0084] 2. Preparation of Polygonum multiflorum No. 1
[0085] Take 10 kg of raw Polygonum multiflorum and slice it into 1 cm thick slices. Wash it with distilled water, dry it at 25°C, sort it by size, add 2.5 kg of black bean juice and mix well, put it in a suitable non-iron container, soak it for 24 hours to allow the black bean juice to be fully absorbed into the slices, steam it under 121°C pressurized steam conditions for 30 hours, then simmer it for 2 hours until the slices appear dark brown, mix it with the remaining black bean juice, dry it at 25°C, put the dried Polygonum multiflorum slices in a sealed bag, ferment them at 25°C for 7 days, and set aside, named as comparative sample 1.
[0086] Application Example 1 Analysis of Microbial Load Quantity
[0087] The raw Polygonum multiflorum, the processed Polygonum multiflorum samples 1 to 3 obtained in the above embodiments 1 to 3, and the processed Polygonum multiflorum A, B, and comparative sample 1 obtained in comparative examples 1 to 3 were respectively counted for microbial counts according to the requirements of the microbial count method (General Rule 1105) in the microbial limit test method for non-sterile products in the 2020 edition of the "Chinese Pharmacopoeia", and the total aerobic bacteria count (TAMC) and the total number of molds and yeasts (TYMC) in all samples were measured.
[0088] a. Preparation of test solution: Take 10 g each of the raw Polygonum multiflorum, the processed Polygonum multiflorum samples 1 to 3 obtained in Examples 1 to 3, and the processed Polygonum multiflorum A, B, and comparative sample 1 obtained in Comparative Examples 1 to 3, grind and homogenize them by aseptic operation, add sterile pH 7.0 peptone liquid culture medium (Qingdao High-tech Park Haibo Biotechnology Co., Ltd., production batch number: 20220407) to 100 mL to prepare a 1:10 (m:V) test solution, and further dilute it with sterile pH 7.0 peptone liquid culture medium to dilute it to 1:100, 1:1000, and 1:10000 dilution levels, respectively.
[0089] b. Determination of bacterial count: Take 1 mL of each dilution level of each sample and place it in a sterile plate with a diameter of 90 mm. Add tryptic soy agar medium to 2 plates and incubate at 33℃ for 5 days. Count the number of colonies on the plates and record it as TAMC. Add Sabouraud dextrose agar medium to the other 2 plates and incubate at 23℃ for 7 days. Count the number of colonies on the plates and record it as TYMC.
[0090] The results of the microbial load test are shown in Table 1 and Figures 1 to 5 As shown:
[0091] Table 1. TAMC and TAYC counts in raw and processed Polygonum multiflorum (unit: cfu / g)
[0092]
[0093] According to the results of microbial counting, the bioburden of raw products and samples prepared by different processing methods was significantly correlated with the processing methods, and TAMC and TAYC increased significantly in processed Polygonum multiflorum I samples 1 to 3.
[0094] Application Example 2 Analysis of Microbial Load Types
[0095] a. Preparation of colonies: Take 10 g each of the raw Polygonum multiflorum, the processed Polygonum multiflorum samples 1 to 3 obtained in Examples 1 to 3, and the processed Polygonum multiflorum A, B, and comparative sample 1 obtained in Comparative Examples 1 to 3, respectively, grind and homogenize using aseptic operation, directly add 100 mL of TSB (Qingdao High-tech Park Haibo Biotechnology Co., Ltd., production batch number: 20220418), and culture at 33 ° C for 24 h, pick 1 loop of culture and streak inoculate it onto TSA (Qingdao High-tech Park Haibo Biotechnology Co., Ltd., production batch number: 20220118) and SDA (Qingdao High-tech Park Haibo Biotechnology Co., Ltd., production batch number: 20230209) plates, and culture at 33 ° C and 23 ° C for 24 to 72 h, pick colonies of different morphologies on the plate, and separate and purify them by streaking on the plate more than 2 times for microbial identification and preservation.
[0096] b. Identification of microbial species: Gram staining of isolated and purified microorganisms was performed on the smear to preliminarily determine the strain type. Bacteria and yeast were identified using VITEKMS and VITEK2compact systems; molds were commissioned to Beijing Nosai Genome Research Center Co., Ltd. for ITS sequence analysis. The samples were amplified by PCR using primers ITS1 / ITS4, the amplified products were purified and recovered by gel cutting, and sequenced using primers ITS1 / ITS4. The sequencing results were compared with Blast on NCBI to complete the identification of the strains. The corresponding strain classification information was obtained and statistically analyzed.
[0097] The results of the microbial load test are shown in Table 2 and Table 3:
[0098] Table 2. Different microbial loads in raw and processed Polygonum multiflorum
[0099]
[0100]
[0101] Table 3. Count results of Aspergillus shevarii in raw and processed Polygonum multiflorum (unit: cfu / g)
[0102]
[0103] The results of the analysis of the types of microbial load showed that the number of bioburdens of the raw product and the samples prepared by different processing methods was significantly related to the processing methods. Samples 1 to 3 of the processed Polygonum multiflorum No. 1 contained 2 strains of Bacillus and 3 strains of mold, and Aspergillus shevarii could be identified. The above colony morphology was shown in Figures 6 to 8 , while Aspergillus serratus was not detected in the raw product of Polygonum multiflorum, processed Polygonum multiflorum A, processed Polygonum multiflorum B, and comparative sample 1, which shows that Aspergillus serratus is a unique strain of processed Polygonum multiflorum No. 1 in this application. And relative to processed Polygonum multiflorum No. 1 sample 2 or processed Polygonum multiflorum No. 1 sample 3, the number of Aspergillus serratus in processed Polygonum multiflorum No. 1 sample 1 in this application is higher, which is more conducive to the retention and growth of Aspergillus serratus. In summary, the present application enhances its fermentation advantage by directional screening and culturing Aspergillus serratus, so that the microorganisms with strict growth conditions such as Aspergillus serratus can effectively grow on Polygonum multiflorum, and obtain a processed Polygonum multiflorum product containing a higher content and unique Aspergillus serratus, which is conducive to promoting the biotransformation in the processing process of Polygonum multiflorum, and broadens the scope of microorganisms accessed by Polygonum multiflorum, which lays the foundation for the research on the potential efficacy of Aspergillus serratus in traditional Chinese medicine, especially in Polygonum multiflorum medicine. In addition, the processed Polygonum multiflorum product prepared by the present application does not contain toxic and harmful microorganisms and their products (such as Aspergillus flavus, Escherichia coli, Salmonella and aflatoxin), which can ensure the safety of the processed Polygonum multiflorum product fermented by the present application. In the processing process and the analysis of the number and type of microbial load, the present application strictly controls the raw materials, steaming process, fermentation process and environment to ensure the stability and controllability of the microorganisms finally detected, and can ensure the repeatability of the number and quality of microorganisms in the processed Polygonum multiflorum product.
[0104] Although the above-mentioned embodiment provides a detailed description of the present application, it is only a part of the embodiments of the present application, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the scope of protection of the present application.
Claims
1. A method for processing Polygonum multiflorum, characterized in that: The following steps are involved: (1) mixing the Polygonum multiflorum slices and the black bean juice, infiltrating them, separating the solid from the liquid, and collecting the infiltrated black bean juice and the infiltrated Polygonum multiflorum slices; (2) steaming the soaked Polygonum multiflorum slices obtained in step (1) to obtain steamed Polygonum multiflorum slices; the steaming temperature is 85 to 100° C.; the steaming time is 22 to 26 hours; (3) stewing the steamed Polygonum multiflorum slices obtained in step (2), mixing the stewed Polygonum multiflorum slices with the soaked black bean juice obtained in step (1), and air-drying the mixed materials to obtain dried Polygonum multiflorum slices; (4) fermenting the dried Polygonum multiflorum slices obtained in step (3) under closed conditions to obtain processed Polygonum multiflorum.
2. The processing method of Radix Polygoni Multiflori according to claim 1, characterized in that: The method for preparing black bean juice in step (1) comprises the following steps: 1) mixing black beans with water for a first cooking, and separating a first liquid component and bean dregs; 2) mixing the okara in step 1) with water for a second cooking, and collecting a second liquid component; 3) combining the first liquid component and the second liquid component to obtain black bean juice.
3. The processing method of Radix Polygoni Multiflori according to claim 2, characterized in that, In step 1), the mass volume ratio of black beans to water is 1 kg: (2.8-3.2) L; the first cooking time is 3.5-4.5 h.
4. The processing method of Radix Polygoni Multiflori according to claim 3, characterized in that: The mass volume ratio of the bean dregs to water in step 2) is 1 kg: (2.8-3.2) L; the second cooking time is 2.5-3.5 h.
5. The processing method of Radix Polygoni Multiflori according to claim 1, characterized in that: The mass ratio of the Polygonum multiflorum slices to the black bean juice in step (1) is 10:(2.1-2.9).
6. The processing method of Radix Polygoni Multiflori according to claim 1, characterized in that: The infiltration time in step (1) is 22 to 26 hours.
7. The processing method of Radix Polygoni Multiflori according to claim 1, characterized in that: The stewing time in step (3) is 1.5 to 2.5 hours.
8. The processing method of Radix Polygoni Multiflori according to claim 1, characterized in that: The fermentation time in step (4) is 6 to 8 days.
9. The processing method of Polygonum multiflorum according to claim 8, characterized in that: The fermentation temperature in step (4) is 20-40°C.
10. The processed Radix Polygoni Multiflori product prepared by the processing method according to any one of claims 1 to 9, characterized in that: The effective viable bacteria count of Aspergillus chevalieri in the processed Polygonum multiflorum product is (250,000) to (310,000) CFU / g.