Microbial solid-state fermented pueraria and preparation method and application thereof
By using solid-state fermentation of Boletus edulis and Aspergillus ryuki, the bitterness and residue problems during the chewing process of kudzu root were solved, the taste was improved, and it showed an effect of improving depressive symptoms in animal models.
Patent Information
- Application Number
- CN202410121583.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2024-01-29
- Publication Date
- 2026-01-23
- Estimated Expiration
- 2044-01-29
AI Technical Summary
Kudzu root has a bitter taste and leaves a lot of residue during chewing, which affects its taste. At the same time, current technology has not been able to effectively improve its depressive symptoms.
Kudzu root was treated with microbial solid-state fermentation using Boletus stalkus and Aspergillus ryuki to promote the hydrolysis of starch and cellulose, neutralize bitterness and produce a specific flavor, improve taste, and at the same time promote the conversion of active ingredients through fungal fermentation, thus improving depressive symptoms.
Fermented kudzu leaves less residue after chewing, improves the taste, and has a unique fruity aroma and umami flavor. It has also significantly improved depressive symptoms in animal experiments.
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Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the technical field of microbial fermentation, and particularly relates to a microbial solid-state fermented puerariae lobatae radix and a preparation method and application thereof. BACKGROUND
[0002] Puerariae lobatae Radix is the dried root of Pueraria lobata (Willd.) Ohwi in the family of Leguminosae, and is a kind of food-medicinal plant. Puerariae lobatae Radix contains functional components such as flavonoids and polysaccharides, and the fibers are rich in cellulose, hemicellulose and lignin. Puerariae lobatae Radix has been applied in the field of chewable food, but puerariae lobatae Radix has a certain bitterness, and the interaction between starch and lignin during chewing will form a lot of residues, which has a certain influence on the chewing taste. The microbial solid-state fermentation technology can effectively improve the taste and enhance the efficacy. SUMMARY
[0003] Therefore, the present application aims to provide a microbial solid-state fermented puerariae lobatae radix and a preparation method thereof, so that the puerariae lobatae radix has less residues after chewing and the taste is improved.
[0004] Another object of the present application is to provide a microbial solid-state fermented puerariae lobatae radix and a preparation method thereof, so that the puerariae lobatae radix has the efficacy of improving the symptoms of depression.
[0005] Another object of the present application is to provide the application of the above-mentioned puerariae lobatae radix in the preparation of puerariae lobatae chewable products.
[0006] Another object of the present application is to provide a chewable product based on the above-mentioned puerariae lobatae radix.
[0007] In order to solve the above technical problems / achieve the above objects or at least partially solve the above technical problems / achieve the above objects, as a first aspect of the present application, a microbial solid-state fermented puerariae lobatae radix is provided, which is formed after solid-state fermentation of Leucopaxillus gregarius and Aspergillus shahii.
[0008] Optionally, the preservation number of the Leucopaxillus gregarius is CGMCC 5.852, and the number of the Aspergillus shahii is CICC41614.
[0009] As a second aspect of the present application, a preparation method of the puerariae lobatae radix is also provided. The mixed bacterial liquid of Leucopaxillus gregarius and Aspergillus shahii is sprayed on the surface of puerariae lobatae radix for solid-state fermentation, and the puerariae lobatae radix is obtained after drying.
[0010] Optionally, the bacterial concentration of the mixed bacterial liquid is (5-20) × 10 7 CFU / mL.
[0011] Optionally, the mixed bacteria solution comprises Leucopaxillus giganteus, Aspergillus luchuensis and culture medium.
[0012] Optionally, the spraying amount of the mixed bacteria solution is 1: (5-20) mL of the mixed bacteria solution to g of the Pueraria lobata.
[0013] Optionally, the temperature of the solid-state fermentation is 24-28℃, and the relative humidity is 70-90%.
[0014] As a third aspect of the present application, the use of the Pueraria lobata in the preparation of a Pueraria lobata chewing product is also provided.
[0015] As a fourth aspect of the present application, a Pueraria lobata chewing product is also provided, which comprises the Pueraria lobata described in the present application.
[0016] The present application uses Leucopaxillus giganteus and Aspergillus luchuensis for solid-state fermentation to treat Pueraria lobata, which can promote the hydrolysis of starch, cellulose and hemicellulose in Pueraria lobata, improve the chewing taste of fiber, reduce residues, produce specific flavors (fruity flavor and umami flavor) and neutralize the bitter taste of Pueraria lobata flavones; at the same time, the fermentation by fungi can promote the transformation of active ingredients in Pueraria lobata and produce some fermentation products. Animal feeding experiments show that the CUMS depression model mice can partially improve the symptoms of depression, and the product can be applied to the development of products for improving depression symptoms. BRIEF DESCRIPTION OF DRAWINGS
[0017] Figure 1 The figure shows the method for constructing a chronic unpredictable mild stress (CUMS) depression model. DETAILED DESCRIPTION
[0018] The present application discloses a kind of Pueraria lobata of microbial solid-state fermentation and its preparation method and application, and those skilled in the art can refer to the content of this paper, and appropriately improve process parameters to realize.It is particularly pointed out that all similar substitutions and changes are obvious to those skilled in the art, and they are all regarded as including in the present application.The product, process and application described in the present application have been described by preferred embodiments, and relevant personnel can obviously make changes or appropriate changes and combinations to the product, process and application described in this paper within the scope of the content, spirit and scope of the present application, to realize and apply the technology of the present application.It is obvious that the described embodiments are part of the embodiments of the present application, not all embodiments.Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor belong to the scope of protection of the present application.
[0019] It should be noted that in this document, relational terms such as "first" and "second," and "step 1" and "step 2," and " (1) " and " (2) " are used solely to distinguish one entity or action from another entity or action, without necessarily requiring or implying any actual relationship or order between such entities or actions. Moreover, the terms "comprises," "comprising," or any other variation thereof are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but can also include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without more limitations, an element defined by an "including" statement does not exclude the existence of additional identical elements in the process, method, article, or apparatus that includes the element. Meanwhile, the embodiments in the present application and the features in the embodiments can be combined with each other without conflict.
[0020] In the first aspect of the present application, the P. ginseng provided by the present application is formed after being subjected to solid-state fermentation by P. ginseng and A. luchuensis. The physiological characteristics of the two strains are used to ferment the active ingredients of P. ginseng, so as to produce new substances and change the original properties of P. ginseng, thereby achieving the intended purpose.
[0021] In some embodiments of the present application, the P. ginseng has a preservation number of CGMCC 5.852, and the Latin name is Suillus granulatus The culture medium is PDA medium, and the optimal culture temperature is 25°C. It can be purchased from the China General Microbiological Culture Collection Center. The A. luchuensis has a number of CICC41614, and the Latin name is Aspergillus luchuensis The spore-producing structure is brownish-brown; the surface is flat; the texture is velvety; the reverse is light grayish yellow, no exudate is produced, no soluble pigment is produced, and the optimal culture temperature is 25°C. The reference use is fermentation of Pu'er tea and production of cellulase. It can be purchased from the China Industrial Microbial Culture Collection Center.
[0022] In the second aspect of the present application, a preparation method of the P. ginseng is simultaneously provided. A mixed bacterial solution of P. ginseng and A. luchuensis is sprayed on the surface of P. ginseng for solid-state fermentation, and the P. ginseng is obtained after drying after the solid-state fermentation.
[0023] In some embodiments of the present application, the P. ginseng is cut into strips before solid-state fermentation, and the specification is 4 cm x 1 cm x 0.5 cm.
[0024] In some embodiments of the present application, the bacterial concentration of the mixed bacterial solution is (5-20) x 10 7 CFU / mL, for example, 5 x 10 7 CFU / mL, 10 x 107 CFU / mL or 20 x 10 7 CFU / mL. Wherein, the ratio of the concentration of the Boletus manicus and Aspergillus awamori is 1:0.1-5, for example, 1:0.1, 1:0.2, 1:0.5:1:1, 1:2, 1:3, 1:4 or 1:5.
[0025] In some embodiments of the present application, the mixed bacteria solution comprises Boletus manicus, Aspergillus awamori and culture medium. Wherein, the culture medium is potato culture medium, which is prepared by 20 g of glucose and 1 L of potato extract (200 g of boiled and filtered potato).
[0026] In some embodiments of the present application, the spraying amount of the mixed bacteria solution is 1: (5-20) g / mL, for example, 1:5, 1:10, 1:15 or 1:20, and the spraying process should ensure that each surface of the kudzu vine is sprayed uniformly.
[0027] In some embodiments of the present application, the temperature of the solid-state fermentation is 24-28℃, the relative humidity is 70-90%, and the fermentation time is generally 5-7 days, which can be adjusted according to actual needs. The mixed bacteria solution can also be sprayed appropriately during the fermentation process.
[0028] The kudzu vine obtained by the solid-state fermentation process according to the present application has a soft hardness, toughness and elasticity, a slightly bitter and astringent taste, a unique fruity aroma and a fresh taste, and less residue after chewing. In addition, after being used to feed a chronic unpredictable mild stress (CUMS) depression mouse model, the mouse tail suspension immobility time is significantly reduced compared with the model group and the comparative example group, indicating that it can improve the despair of depressed mice and help to improve the symptoms of depression. Based on these experimental results, the present application proposes the use of the kudzu vine in the preparation of a kudzu vine chewing product in the third aspect.
[0029] In the fourth aspect of the present application, according to the application scope of the kudzu vine, a kudzu vine chewing product is provided, which comprises the kudzu vine described in the present application, and other suitable efficacy ingredients and food additives can also be added according to needs, and the adding methods include but are not limited to soaking and smearing.
[0030] In each group of comparative experiments provided in the present application, unless otherwise specified, the experimental conditions, materials, etc. remain the same except for the differences indicated in each group, so as to have comparability. The experimental materials and reagents used in the examples can be obtained from the market unless otherwise specified. The point handle sticky cap boletus used in the specific examples is a strain numbered CGMCC 5.852 purchased from the China General Microbiological Culture Collection Center, and the Aspergillus shivajii is a strain numbered CICC41614 purchased from the China Industrial Microbial Culture Collection Center.
[0031] The following further illustrates a microorganism solid-state fermented pueraria and a preparation method and application thereof provided in the present application.
[0032] Example 1:
[0033] The dried pueraria was cut into small strips of 4 cm x 1 cm x 0.5 cm, and 10 8 CFU / mL concentration of mixed bacteria liquid of the point handle sticky cap boletus and the Aspergillus shivajii (the concentration ratio of the point handle sticky cap boletus to the Aspergillus shivajii was 1:0.2), the fermentation temperature was 25°C, the humidity was 70-90%, and the fermentation time was 7 days. After the fermentation was completed, hot air drying was performed at 80°C.
[0034] Example 2:
[0035] The dried pueraria was cut into small strips of 4 cm x 1 cm x 0.5 cm, and 10 8 CFU / mL concentration of mixed bacteria liquid of the point handle sticky cap boletus and the Aspergillus shivajii (the concentration ratio of the point handle sticky cap boletus to the Aspergillus shivajii was 1:2), the fermentation temperature was 25°C, the humidity was 70-90%, and the fermentation time was 7 days. After the fermentation was completed, hot air drying was performed at 80°C.
[0036] Comparative Example 1:
[0037] The dried pueraria was cut into small strips of 4 cm x 1 cm x 0.5 cm, and 10
[0038] Comparative Example 2:
[0039] The dried pueraria was cut into small strips of 4 cm x 1 cm x 0.5 cm, and 10 8 CFU / mL concentration of mixed bacteria liquid of the point handle sticky cap boletus and the Aspergillus shivajii (the concentration ratio of the point handle sticky cap boletus to the Aspergillus shivajii was 1:2), the fermentation temperature was 25°C, the humidity was 70-90%, and the fermentation time was 7 days. After the fermentation was completed, hot air drying was performed at 80°C.
[0040] Among them, Bacillus subtilis and Kluyveromyces lactis are strains in the prior application CN02410075904.4, the number of Bacillus subtilis is CICC 21342, and the number of Kluyveromyces lactis is CICC 32410, which are purchased from China Industrial Microbial Culture Collection Center.
[0041] Experimental example:
[0042] (1) Chewing experiment
[0043] The Puerariae radix strips of each example, the comparative example, and the Puerariae radix strip without any treatment were chewed for three minutes by the same person. After completing one treatment group, the person rinsed his mouth with water and rested for 30 minutes. Before performing the next group, the person needed to rinse his mouth again, and then made a taste evaluation comparison. Table 1 is shown below.
[0044] Table 1
[0045]
[0046] As can be seen from Table 1, the Puerariae radix without any treatment is extremely hard and difficult to chew, and is bitter and astringent, which is consistent with the characteristics of natural Puerariae radix. Comparative example 1 has no obvious difference in hardness and taste from the Puerariae radix without any treatment because it is only sprayed with water. However, the examples 1, 2, and comparative example 2, which are substantially treated (fermented by microbial strains), can change the characteristics of Puerariae radix, making the hardness soft and the bitter and astringent taste less. However, it is found that different microbial strains have some differences in the hardness of the taste. Compared with Bacillus subtilis and Kluyveromyces lactis, the hardness of the Puerariae radix fermented by the Boletus piperatus and Aspergillus shahii in the present application is not only soft but also has toughness and elasticity. The taste is also increased with unique fruity and fresh flavors, and the debris is also small.
[0047] (2) Mouse animal model experiment
[0048] A total of 72 SPF male (Kunming) mice were divided into 6 groups, 12 in each group (3 cages), and were normally given drinking water and standard maintenance mouse feed. The groups are as follows:
[0049] The blank control group was normally given drinking water and standard maintenance mouse feed throughout the experiment.
[0050] The model control group was normally given drinking water and standard maintenance mouse feed for one month, and then a chronic unpredictable mild stress (CUMS) depression model was constructed according to the method shown in the following table. Figure 1
[0051] The example 1 group was normally given drinking water, standard maintenance mouse feed, and one Puerariae radix strip (cut and fed, once a day, 50 g / kg body weight) for one month, and then a chronic unpredictable mild stress (CUMS) depression model was constructed according to the method shown in the following table. Figure 1 The method shown builds a chronic unpredictable mild stress (CUMS) depression model.
[0052] Example 2 group: normal administration of drinking water, standard maintenance mouse feed and one root of the ginseng root of Example 2 (cut and fed, once a day, 50 g / kg body weight each time) for one month, and then simultaneously according to Figure 1 The method shown builds a chronic unpredictable mild stress (CUMS) depression model.
[0053] Comparative Example 1 group: normal administration of drinking water, standard maintenance mouse feed and one root of the ginseng root of Comparative Example 1 (cut and fed, once a day, 50 g / kg body weight each time) for one month, and then simultaneously according to Figure 1 The method shown builds a chronic unpredictable mild stress (CUMS) depression model.
[0054] Comparative Example 2 group: normal administration of drinking water, standard maintenance mouse feed and one root of the ginseng root of Comparative Example 2 (cut and fed, once a day, 50 g / kg body weight each time) for one month, and then simultaneously according to Figure 1 The method shown builds a chronic unpredictable mild stress (CUMS) depression model.
[0055] After 4 weeks of treatment according to the CUMS method, the tail suspension test was performed: the mice were placed in the test room for 30 min before the test. The skin about 2 cm from the tail tip of the mouse was stuck with tape, and it was in a suspended state. The test was performed for 5 min, and the immobile time of the mouse in the last 3 min was recorded. The results are shown in Table 2 below;
[0056] Table 2
[0057]
[0058] As can be seen from Table 2, the immobile time of the mice in the model control group was significantly prolonged compared with the blank control group, indicating that the model was successfully constructed according to the CUMS method; except for Comparative Example 1 group, the other three groups could significantly reduce the immobile time of the mice, indicating that the ginseng root fermented by the microbial strain could produce active ingredients that could improve the despair of depressed mice, but the ginseng root prepared in the present application was the best.
[0059] The above description is merely specific embodiments of the present application, enabling those skilled in the art to understand or implement the present application. Various modifications to these embodiments will be apparent to those skilled in the art, and the general principles defined herein can be implemented in other embodiments without departing from the spirit or scope of the present application. Therefore, the present application will not be limited to these embodiments shown herein, but will conform to the widest scope consistent with the principles and novel features applied herein.
Claims
1. A kudzu root fermented by microbial solid-state fermentation, characterized in that, It is formed by solid-state fermentation of *Boletus styracifolium* and *Aspergillus ryukyuensis*. In this process, a mixed bacterial solution including Boletus stoloniferus and Aspergillus ryukyu was sprayed onto the surface of kudzu root for solid-state fermentation, and the kudzu root was obtained by drying after solid-state fermentation. The bacterial concentration of the mixed bacterial solution is (5-20)×10 7 CFU / mL; The spraying amount of the mixed bacterial solution is based on the ratio of mixed bacterial solution volume (mL) to kudzu root weight (g) = 1: (5-20). The solid-state fermentation temperature is 24-28℃, and the relative humidity is 70-90%. Boletus edulis: The bacterial concentration ratio of Aspergillus ryukyu is 1:0.1-5; Fermentation time is 5-7 days; The preservation number of the *Boletus edulis* is CGMCC 5.852; The Aspergillus ryukyu species is designated CICC41614.
2. The use of kudzu root as described in claim 1 in the preparation of kudzu root chewable products.
3. A kudzu root chewable product, characterized in that, Includes the kudzu root as described in claim 1.
Citation Information
Patent Citations
Polyphenol-rich, tea-flavored soy sauce and brewing method thereof
CN111150045A