Ganoderma lucidum fermentation liquor capable of improving content of total triterpenes and improving immunity as well as preparation method and application of ganoderma lucidum fermentation liquor

Abstract

The invention discloses a ganoderma lucidum fermentation liquid capable of improving the content of total triterpenes and immunity and a preparation method and application thereof, the preparation method comprises the following steps: heating a ganoderma lucidum liquid, and carrying out hot water extraction to obtain a ganoderma lucidum extracting liquid; heating the lucid ganoderma extracting solution to be boiled, after sterilization is finished, after the lucid ganoderma liquid is cooled, adding lactic acid bacteria powder in a specific proportion for fermentation to obtain lucid ganoderma fermentation liquid; and carrying out solid-liquid separation on the fermentation liquor to obtain fermentation supernate, and sterilizing the ganoderma lucidum fermentation liquor to obtain the sterilized ganoderma lucidum fermentation liquor. According to the preparation method disclosed by the invention, the content of total triterpenes in the ganoderma lucidum fermentation liquor is increased, the immunomodulatory activity of the ganoderma lucidum product can be enhanced, and the ganoderma lucidum product has relatively high effects of resisting oxidation, improving the activity of phagocytes, improving the activity of acid phosphatase, improving the immunity and the like, and has a wide application prospect and market potential.

Description

Technical Field

[0001] The present invention relates to the technical field of ganoderma lucidum fermentation product processing, and in particular to a ganoderma lucidum fermentation liquid with improved total triterpenoid content and immunity, and a preparation method and application thereof. Background Art

[0002] Ganoderma lucidum, listed in Volume 1 of the Pharmacopoeia of the People's Republic of China, is the dried fruiting body of Ganoderma lucidum (Leyss. exFr.) Karst. or Ganoderma sinense (Zhao, Xu et Zhang). As a traditional Chinese medicinal material, Ganoderma lucidum has attracted considerable attention for its rich bioactive components and pharmacological effects. The main active ingredients in Ganoderma lucidum include triterpenes, polysaccharides, and sterols. Triterpenes, due to their unique pharmacological activities, such as enhancing immunity, anti-oxidation, and anti-inflammatory properties, have become an important indicator of Ganoderma lucidum quality evaluation.

[0003] However, traditional methods for extracting Ganoderma triterpenes, such as hot water extraction and ethanol extraction, have problems such as low extraction efficiency, high cost, and possible adverse effects on the structure of Ganoderma triterpenes.

[0004] In recent years, with the continuous development of microbial fermentation technology, its application in the field of traditional Chinese medicine extraction has become increasingly widespread. Microbial fermentation can destroy plant cell walls, increasing the release rate of active ingredients. Furthermore, enzymes produced during the fermentation process can biotransform traditional Chinese medicine components, enhancing their bioactivity. Therefore, using microbial fermentation technology to improve the extraction rate and bioactivity of Ganoderma lucidum triterpenes has become a research hotspot in the field of Ganoderma lucidum deep processing.

[0005] However, there are still some problems in the existing Ganoderma lucidum fermentation technology, such as the selection of fermentation strains and the optimization of fermentation conditions, which will affect the quality and biological activity of Ganoderma lucidum fermentation liquid. Summary of the Invention

[0006] The purpose of the present invention is to provide a Ganoderma lucidum fermentation liquid with improved total triterpenoid content and immunity, as well as a preparation method and application thereof, which can improve the extraction rate of total triterpenoids in Ganoderma lucidum products, enhance the immunomodulatory activity of Ganoderma lucidum products, and improve the economic value and market potential of Ganoderma lucidum products.

[0007] To achieve the above object, the present invention provides a method for preparing a Ganoderma lucidum fermentation broth with improved total triterpenoid content and immunity, comprising the following steps:

[0008] Step 1: Pretreatment of raw materials: Ganoderma lucidum is crushed until there are no lumps, and then Ganoderma lucidum powder is mixed with deionized water at a mass ratio of 1:20 to 30 to obtain a Ganoderma lucidum mixed solution;

[0009] Step 2: Extraction: extracting the Ganoderma lucidum mixture with hot water to obtain Ganoderma lucidum extract;

[0010] Step 3: Sterilization before fermentation: Heat the Ganoderma lucidum extract to boiling for sterilization, and then cool it down after sterilization;

[0011] Step 4, fermentation: adding 0.005-0.02% of lactic acid bacteria and 2-5% of oligofructose to the cooled Ganoderma lucidum extract at a temperature of 36-38° C. for 18-48 hours to obtain Ganoderma lucidum fermentation liquid;

[0012] The added lactic acid bacteria are a mixture of one or more of Lactobacillus plantarum MPs-68, Lactobacillus rhamnosus YYS-B2 and Lactobacillus paracasei YYS-UR;

[0013] Among them, the deposit number of Lactobacillus plantarum MPs-68 is CGMCC No.31976; the deposit number of Lactobacillus rhamnosus is CGMCC No.28164; the deposit number of Lactobacillus paracasei YYS-UR is CGMCC No.28533;

[0014] Step 5, solid-liquid separation: performing solid-liquid separation on the Ganoderma lucidum fermentation liquid to obtain Ganoderma lucidum fermentation supernatant;

[0015] Step 6: Sterilization after fermentation: sterilize the Ganoderma lucidum fermentation supernatant to obtain a sterilized Ganoderma lucidum fermentation liquid.

[0016] Furthermore, in the hot water extraction process of step 2, the Ganoderma lucidum mixture is extracted with water at 90-100° C. for 160-240 minutes;

[0017] Step 3: Sterilization: Heat the Ganoderma lucidum extract to boiling and keep it for 20 to 30 minutes for sterilization. After sterilization, cool the Ganoderma lucidum extract to 37°C ± 2°C.

[0018] Furthermore, during the fermentation process in step 4, the lactic acid bacteria species are composed of any two of Lactobacillus plantarum MPs-68, Lactobacillus rhamnosus YYS-B2, and Lactobacillus paracasei YYS-UR;

[0019] When the lactic acid bacteria powder consists of Lactobacillus plantarum MPs-68 and Lactobacillus rhamnosus YYS-B2, the mass ratio of the two is 1-3:1-4;

[0020] When the lactic acid bacteria powder consists of Lactobacillus plantarum MPs-68 and Lactobacillus paracasei YYS-UR, the mass ratio of the two is 1-3:1-5;

[0021] When the lactic acid bacteria powder consists of Lactobacillus rhamnosus YYS-B2 and Lactobacillus paracasei YYS-UR, the mass ratio of the two is 1-4:1-5;

[0022] The number of viable bacteria of the Lactobacillus plantarum MPs-68 powder is 1 to 5×10 11 CFU / g;

[0023] The number of viable bacteria of the Lactobacillus rhamnosus YYS-B2 bacterial powder is 1 to 5×10 11 CFU / g;

[0024] The viable bacteria count of the Lactobacillus paracasei YYS-UR bacterial powder is 1 to 5×10 11 CFU / g.

[0025] Furthermore, during the fermentation process in step 4, the lactic acid bacteria powder is composed of Lactobacillus rhamnosus YYS-B2 and Lactobacillus paracasei YYS-UR, and the mass ratio of the two is 1-3:2-4;

[0026] Alternatively, the mass ratio of the two is 1.5-2.5:2.5-3.5.

[0027] Furthermore, during the fermentation process in step 4, the lactic acid bacteria strains are composed of Lactobacillus plantarum MPs-68, Lactobacillus rhamnosus YYS-B2 and Lactobacillus paracasei YYS-UR, and the mass ratio of the three is 1-3:1-4:1-5;

[0028] The number of viable bacteria of the Lactobacillus plantarum MPs-68 powder is 1 to 5×10 11 CFU / g;

[0029] The number of viable bacteria of the Lactobacillus rhamnosus YYS-B2 bacterial powder is 1 to 5×10 11 CFU / g;

[0030] The viable bacteria count of the Lactobacillus paracasei YYS-UR bacterial powder is 1 to 5×10 11 CFU / g.

[0031] Furthermore, during the fermentation process in step 4, the lactic acid bacteria strains are composed of Lactobacillus plantarum MPs-68, Lactobacillus rhamnosus YYS-B2 and Lactobacillus paracasei YYS-UR, and the mass ratio of the three is 1.5-2.5:1.5-3.5:1.5-4.5.

[0032] Furthermore, the Lactobacillus plantarum MPs-68, Lactobacillus rhamnosus YYS-B2 and Lactobacillus paracasei YYS-UR are provided in the form of bacterial powder or bacterial liquid.

[0033] Furthermore, in the solid-liquid separation process of step 5, the Ganoderma lucidum fermentation liquid is first filtered using a 300-mesh sieve, and then centrifuged using a centrifuge to separate the solid and liquid. The centrifugation condition is 6000 r / min for 10 min, and then the Ganoderma lucidum fermentation supernatant is collected;

[0034] Step 6: Sterilization after fermentation: sterilization conditions are 100-110°C, 20-30 minutes.

[0035] A ganoderma lucidum fermentation liquid with improved total triterpenoid content and immunity is prepared by the above-mentioned preparation method, and the total triterpenoid content of the ganoderma lucidum fermentation liquid reaches above 0.2g / 100mL.

[0036] A functional product containing Ganoderma lucidum fermentation liquid, whose ingredients include the Ganoderma lucidum fermentation liquid with improved total triterpenoid content and immunity; or its ingredients include the Ganoderma lucidum fermentation liquid with improved total triterpenoid content and immunity prepared by the above preparation method.

[0037] After adopting the above solution, the beneficial effects of the present invention are:

[0038] (1) Significantly improve the total triterpene content: The present invention ferments the Ganoderma lucidum water extract by using specific Lactobacillus plantarum MPs-68, Lactobacillus rhamnosus YYS-B2, Lactobacillus paracasei YYS-UR and mixed strains to obtain a Ganoderma lucidum fermentation liquid with a high total triterpene content, and the total triterpene content reaches more than 0.2g / 100mL.

[0039] The present invention further improves the total triterpene content in the Ganoderma lucidum fermentation liquid by selecting a mixed bacteria of Lactobacillus rhamnosus YYS-B2 and Lactobacillus paracasei YYS-UR and fermenting the Ganoderma lucidum liquid in a specific ratio, and the total triterpene content reaches 0.24g / 100mL.

[0040] (2) Immunity Enhancement: The fermented Ganoderma lucidum broth prepared by the present invention has antioxidant properties, improves phagocyte viability, and enhances acid phosphatase activity. It can be applied to various functional products (such as functional foods, cosmetics, pharmaceuticals, and health supplements) to enhance the body's immunity. Experimental data show that the fermented Ganoderma lucidum broth can increase the secretion of NO, IL-6, and TNF-α in RAW264.7 cells. These cytokines play an important role in immune regulation.

[0041] (3) Improved taste and flavor: The fermented Ganoderma lucidum broth prepared by the present invention has a rich aroma, a mellow taste, and a balance of sourness and sweetness, and is suitable as a raw material for functional foods.

[0042] (4) Simple and easy process: The method of the present invention uses simple operations such as extraction, fermentation, and filtration to produce the desired Ganoderma lucidum fermentation liquid. The process is simple and easy to operate, and the equipment requirements are relatively low, making it suitable for large-scale production. In addition, no organic solvents are added during the production process, which can be effectively utilized in the food industry. The fermented Ganoderma lucidum liquid has broad application prospects and market potential. DETAILED DESCRIPTION

[0043] The present invention is described in detail below with reference to specific embodiments.

[0044] The strains used in this invention, Lactobacillus plantarum MPs-68, Lactobacillus rhamnosus YYS-B2, and Lactobacillus paracasei YYS-UR, are all publicly available strains, preserved and maintained at a depository. A skilled artisan can obtain these strains from the depository using their depository numbers. Therefore, this application does not involve strain preservation, and proof of survival of these strains is not required.

[0045] The present invention provides a Ganoderma lucidum fermentation liquid for improving total triterpenoid content and immunity, and a preparation method and application thereof, comprising the following steps:

[0046] Step 1: Raw material pretreatment:

[0047] Ganoderma lucidum is crushed until there are no lumps, and then Ganoderma lucidum powder is mixed with deionized water at a mass ratio of 1:20-30 to obtain a Ganoderma lucidum mixed solution;

[0048] Step 2: Extraction:

[0049] extracting the Ganoderma lucidum mixture in hot water at 90-100° C. for 160-240 minutes to obtain a Ganoderma lucidum extract;

[0050] Step 3: Sterilization before fermentation:

[0051] Heat the Ganoderma lucidum extract to boiling and keep it for 20 to 30 minutes for sterilization. After sterilization, cool the Ganoderma lucidum extract to 37℃±2℃;

[0052] Step 4: Fermentation:

[0053] The cooled Ganoderma lucidum extract is added with 0.005-0.02% of lactic acid bacteria and 2-5% of oligofructose in the liquid system for fermentation at a temperature of 36-38° C. for 18-48 hours to obtain Ganoderma lucidum fermentation liquid.

[0054] The added lactic acid bacteria are a mixture of one or more of Lactobacillus plantarum MPs-68, Lactobacillus rhamnosus YYS-B2, and Lactobacillus paracasei YYS-UR. This process utilizes specific lactic acid bacteria to ferment Ganoderma lucidum. Enzymes released during fermentation can be used to break down the cell walls of the Ganoderma lucidum, thereby improving the extraction rate of Ganoderma lucidum triterpenes and enhancing the biological activity of the Ganoderma lucidum liquid.

[0055] The Lactobacillus plantarum strain MPs-68 used was deposited at the General Microbiology Center of the China Culture Collection Administration, with a deposit number of CGMCC No. 31976. The deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, and the deposit date is September 6, 2024.

[0056] The Lactobacillus rhamnosus YYS-B2 used is deposited at the General Microbiology Center of the China Culture Collection Administration, with a deposit number of CGMCC No. 28164. The deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, and the deposit date is August 14, 2023.

[0057] The Lactobacillus paracasei strain YYS-UR was deposited at the General Microbiology Center of the China Culture Collection Administration under the CGMCC No. 28533. The deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, and the deposit date is September 25, 2023.

[0058] In this protocol, Lactobacillus plantarum MPs-68, Lactobacillus paracasei YYS-K1, Lactobacillus fermentans B153, Lactobacillus rhamnosus YYS-B2, and Lactobacillus paracasei YYS-UR are provided in the form of bacterial powder or bacterial solution. When provided in the form of bacterial powder,

[0059] The number of viable bacteria of the Lactobacillus plantarum MPs-68 powder is 1 to 5×10 11 CFU / g;

[0060] The number of viable bacteria of the Lactobacillus rhamnosus YYS-B2 bacterial powder is 1 to 5×10 11 CFU / g;

[0061] The viable bacteria count of the Lactobacillus paracasei YYS-UR bacterial powder is 1 to 5×1011 CFU / g.

[0062] The lactic acid bacteria powder is composed of any two of the lactic acid bacteria species selected from Lactobacillus plantarum MPs-68, Lactobacillus rhamnosus YYS-B2, and Lactobacillus paracasei YYS-UR: when the lactic acid bacteria powder is composed of Lactobacillus plantarum MPs-68 and Lactobacillus rhamnosus YYS-B2, the mass ratio of the two is 1-3:1-4; when the lactic acid bacteria powder is composed of Lactobacillus plantarum MPs-68 and Lactobacillus paracasei YYS-UR, the mass ratio of the two is 1-3:1-5; when the lactic acid bacteria powder is composed of Lactobacillus rhamnosus YYS-B2 and Lactobacillus paracasei YYS-UR, the mass ratio of the two is 1-4:1-5;

[0063] Furthermore, when the lactic acid bacteria powder consists of Lactobacillus rhamnosus YYS-B2 and Lactobacillus paracasei YYS-UR, the mass ratio of the two is 1-3:2-4.

[0064] Furthermore, when the lactic acid bacteria powder consists of Lactobacillus rhamnosus YYS-B2 and Lactobacillus paracasei YYS-UR, the mass ratio of the two is 1.5-2.5:2.5-3.5.

[0065] The lactic acid bacteria powder consists of Lactobacillus plantarum MPs-68, Lactobacillus rhamnosus YYS-B2 and Lactobacillus paracasei YYS-UR, and the mass ratio of the three is 1-3:1-4:1-5.

[0066] Furthermore, the lactic acid bacteria powder consists of Lactobacillus plantarum MPs-68, Lactobacillus rhamnosus YYS-B2 and Lactobacillus paracasei YYS-UR, and the mass ratio of the three is 1.5-2.5:1.5-3.5:1.5-4.5.

[0067] Step 5, solid-liquid separation: first use a 300-mesh sieve to perform preliminary filtration on the Ganoderma lucidum fermentation liquid, and then use a centrifuge to centrifuge to separate the solid and liquid. The centrifugation condition is 6000r / min for 10min, and then collect the Ganoderma lucidum fermentation supernatant.

[0068] Step 6: Sterilization after fermentation: sterilize the Ganoderma lucidum fermentation supernatant at 100-110° C. for 20-30 min to obtain a sterilized Ganoderma lucidum fermentation liquid.

[0069] The ganoderma lucidum fermentation liquid prepared by the preparation method of the present invention has a high total triterpenoid content and also has the effects of anti-oxidation, improving phagocyte activity, improving acid phosphatase activity, improving immunity and the like.

[0070] Since the fermented Ganoderma lucidum liquid has antioxidant, phagocyte activity enhancement, immunity enhancement and other effects, it can be used in various functional products, which helps to improve the body's immunity from multiple angles and improve people's quality of life.

[0071] The present application provides the following examples and comparative examples to verify the effectiveness of the present application scheme:

[0072] Examples 1 to 7 are about the selection of different strains; Examples 8 to 10 are about the selection of different fermentation times; Comparative Examples 5 to 13 are about the selection of different carbon source addition amounts; Comparative Example 1 is unfermented Ganoderma lucidum liquid, and Comparative Examples 2 to 4 are fermentation liquids of commercially available strains.

[0073] <Example 1>

[0074] Ganoderma lucidum fermentation liquid with improved total triterpenoid content and immunity, and its preparation method and application:

[0075] (1) Ganoderma lucidum was ground into powder using a grinder. 100 g of Ganoderma lucidum powder was weighed and added into 3000 mL of deionized water at 95°C (the ratio of material to water was 1:30). The mixture was stirred and extracted for 3 h. The mixture was then extracted with hot water to prepare Ganoderma lucidum extract.

[0076] (2) Boil the Ganoderma lucidum extract for 25 minutes to sterilize it. After the sterilization is completed, cool the Ganoderma lucidum extract to 37±2℃.

[0077] (3) Weigh 0.01% of the Lactobacillus plantarum MPs-68 powder in the liquid system and inoculate it into the cooled Ganoderma lucidum extract. The number of viable bacteria of the Lactobacillus plantarum MPs-68 powder is 1 to 5×10 11 CFU / g, and then 3% oligofructose was weighed and added into the Ganoderma lucidum extract, and fermented at 36-38°C for 18h to obtain Ganoderma lucidum fermentation liquid.

[0078] (4) The fermented liquor of Ganoderma lucidum was subjected to solid-liquid separation after fermentation. The fermented liquor was initially filtered using a 300-mesh filter cloth, and then centrifuged at 6000 r / min for 10 min. The supernatant of the fermented liquor was collected after centrifugation.

[0079] (5) The collected Ganoderma lucidum fermentation supernatant was sterilized at 105° C. for 25 min to obtain a sterilized Ganoderma lucidum fermentation liquid.

[0080] <Example 2>

[0081] The difference between this embodiment and embodiment 1 is that an equal amount of Lactobacillus rhamnosus YYS-B2 is used to replace Lactobacillus plantarum MPs-68, and the remaining operations and processes are the same as those in embodiment 1.

[0082] <Example 3>

[0083] The difference between this embodiment and embodiment 1 is that an equal amount of Lactobacillus paracasei YYS-UR is used to replace Lactobacillus plantarum MPs-68, and the remaining operations and processes are the same as those in embodiment 1.

[0084] <Example 4>

[0085] In this example, Ganoderma lucidum fermentation liquid was prepared with reference to Example 1, and the same mass of lactic acid bacteria powder as in Example 1 was added. However, the lactic acid bacteria powder was a mixed bacterial powder of Lactobacillus plantarum MPs-68 and Lactobacillus rhamnosus YYS-B2, and the mass ratio of the two was 1:2.

[0086] <Example 5>

[0087] In this example, Ganoderma lucidum fermentation liquid was prepared with reference to Example 1, and the same mass of lactic acid bacteria powder as in Example 1 was added. However, the lactic acid bacteria powder was a mixed bacterial powder of Lactobacillus plantarum MPs-68 and Lactobacillus paracasei YYS-UR, and the mass ratio of the two was 1:3.

[0088] <Example 6>

[0089] In this example, Ganoderma lucidum fermentation liquid was prepared with reference to Example 1, and the same mass of lactic acid bacteria powder as in Example 1 was added. However, the lactic acid bacteria powder was a mixed bacterial powder of Lactobacillus rhamnosus YYS-B2 and Lactobacillus paracasei YYS-UR, and the mass ratio of the two was as follows:

[0090] Example 6-1: YYS-B2: YYS-UR = 2:3,

[0091] Example 6-2: YYS-B2: YYS-UR = 1:5,

[0092] Example 6-3: YYS-B2: YYS-UR = 1:1,

[0093] Example 6-4: YYS-B2:YYS-UR=3:1,

[0094] Example 6-5: YYS-B2:YYS-UR=5:1.

[0095] <Example 7>

[0096] In this example, Ganoderma lucidum fermentation liquid was prepared with reference to Example 1, and the same mass of lactic acid bacteria powder as that in Example 1 was added. The lactic acid bacteria powder was a mixed bacterial powder of Lactobacillus plantarum MPs-68, Lactobacillus YYS-B2, and Lactobacillus paracasei YYS-UR, with a mass ratio of 1:2:3.

[0097] Examples 8 to 13 all used the same mixed strains as Example 6.

[0098] <Example 8>

[0099] The difference between this example and Example 6-1 is that after the bacterial strain is introduced, the fermentation time is extended to 24 hours (18 hours in Example 6-1). The remaining operations and processes are the same as in Example 6-1.

[0100] <Example 9>

[0101] The difference between this example and Example 6-1 is that after the bacterial strain is introduced, the fermentation time is extended to 36 hours. The remaining operations and processes are the same as those in Example 6-1.

[0102] <Example 10>

[0103] The difference between this example and Example 6-1 is that after the bacterial strain is introduced, the fermentation time is extended to 48 hours. The remaining operations and processes are the same as those in Example 6-1.

[0104] <Example 11>

[0105] The difference between this example and Example 6-1 is that after the bacteria are inoculated, the amount of oligofructose added is 2% of the liquid system. The rest of the operations and processes are the same as those in Example 6-1.

[0106] <Example 12>

[0107] The difference between this example and Example 6-1 is that after the bacteria are inoculated, the amount of oligofructose added is 4% of the liquid system. The rest of the operations and processes are the same as those in Example 6-1.

[0108] <Example 13>

[0109] The difference between this example and Example 6-1 is that after the bacteria are inoculated, the amount of oligofructose added is 5% of the liquid system. The rest of the operations and processes are the same as those in Example 6-1.

[0110] Comparative Example 1

[0111] In the preparation method of the Ganoderma lucidum product of this comparative example, no fermentation treatment was performed, and the specific steps were as follows:

[0112] (1) Ganoderma lucidum was ground into powder using a grinder. 100 g of Ganoderma lucidum powder was weighed and added into 3000 mL of deionized water at 95°C (the ratio of material to water was 1:30). The mixture was stirred and extracted for 3 h to prepare Ganoderma lucidum extract.

[0113] (2) Boil the Ganoderma lucidum extract for 25 minutes to sterilize. After sterilization, cool the Ganoderma lucidum extract to 35°C to 39°C.

[0114] (3) The Ganoderma lucidum extract was subjected to solid-liquid separation. The extract was initially filtered using a 300-mesh filter cloth, and the filtered extract was centrifuged at 6000 r / min for 10 min. The supernatant after centrifugation was collected.

[0115] (4) The collected Ganoderma lucidum extract supernatant was sterilized at 105°C for 25 minutes to obtain Ganoderma lucidum extract.

[0116] Comparative Example 2

[0117] The difference between this comparative example and Example 1 is that an equal amount of commercially available F-19 Lactobacillus paracasei is used to replace Lactobacillus plantarum MPs-68, and the remaining operations are the same as those in Example 1.

[0118] Comparative Example 3

[0119] The difference between this comparative example and Example 6-1 is that the lactic acid bacteria powder is a mixed bacterial powder of Lactobacillus rhamnosus YYS-B2 and Lactobacillus paracasei F-19, and the mass ratio of the two is 2:3. The other operations are the same as those in Example 1.

[0120] Comparative Example 4

[0121] The difference between this comparative example and Example 6-1 is that the lactic acid bacteria powder is a mixed bacterial powder of Lactobacillus paracasei F-19 and Lactobacillus paracasei YYS-UR, and the mass ratio of the two is 2:3. The other operations are the same as those in Example 1.

[0122] Comparative Example 5

[0123] The difference between this comparative example and Example 6-1 is that after the bacteria are inoculated, the amount of oligofructose added is 1% of the liquid system (3% in Example 6-1). The rest of the operations and processes are the same as in Example 6-1.

[0124] The performance tests were conducted on the fermented Ganoderma lucidum liquids obtained in the above examples and comparative examples:

[0125] Result Example 1: Determination of total triterpenoid content in fermented Ganoderma lucidum liquid. The detection method refers to the "Agricultural Industry Standard of the People's Republic of China" - NY / T 3676-2020 Determination of total triterpenoid content in Ganoderma lucidum spectrophotometric method:

[0126] Table 1 Content and improvement rate of total triterpenes in Examples and Comparative Examples (%)

[0127] project Total triterpenoid content (g100mL) Improvement rate (%) Example 1 0.216±0.001 217.6 Example 2 0.227±0.001 233.8 Example 3 0.211±0.002 210.3 Example 4 0.228±0.002 235.3 Example 5 0.230±0.003 238.2 Example 6-1 0.240±0.001 252.9 Example 6-2 0.234±0.002 244.1 Example 6-3 0.231±0.001 239.7 Example 6-4 0.235±0.003 245.6 Example 6-5 0.233±0.002 242.6 Example 7 0.222±0.001 226.5 Example 8 0.222±0.002 226.5 Example 9 0.217±0.001 219.1 Example 10 0.204±0.002 200.0 Example 11 0.188±0.003 176.5 Example 12 0.259±0.001 280.9 Example 13 0.244±0.002 258.8 Comparative Example 1 0.068±0.002 - Comparative Example 2 0.164±0.002 141.2 Comparative Example 3 0.225±0.003 230.9 Comparative Example 4 0.223±0.001 217.9 Comparative Example 5 0.103±0.002 51.5

[0128] Effects of fermentation:

[0129] As shown in Table 1, the total triterpene content of the Ganoderma lucidum fermentation liquid obtained by the strain fermentation method in the embodiment of the present invention is much higher than the total triterpene content of the Ganoderma lucidum liquid without fermentation treatment in Comparative Example 1, indicating that the preparation method of the present invention can effectively increase the total triterpene content of the Ganoderma lucidum liquid.

[0130] The impact of a single bacterial species:

[0131] Comparative Examples 1 to 3 show that when fermented with one of Lactobacillus plantarum MPs-68, Lactobacillus rhamnosus YYS-B2, and Lactobacillus paracasei YYS-UR, the highest increase in total triterpene content in the Ganoderma lucidum fermentation broth was achieved with Lactobacillus rhamnosus YYS-B2 (Example 2). Lactobacillus rhamnosus YYS-B2 releases specific enzymes during the fermentation process that effectively destroy the Ganoderma lucidum cell wall, thereby more fully extracting the triterpene components from the Ganoderma lucidum.

[0132] Comparative Example 2 uses commercially available F-19 Lactobacillus paracasei for fermentation. In comparison, the total triterpene content of the Ganoderma lucidum fermentation liquid in the examples of the present invention is significantly higher than the total triterpene content of the Ganoderma lucidum liquid in Comparative Example 2. Therefore, it can be shown that the strains selected by the present invention have a significant effect on increasing the total triterpene content in the Ganoderma lucidum liquid.

[0133] Effects of mixed strains:

[0134] Comparing Examples 4 to 6 with Examples 1 to 3, it can be seen that when any two of the three strains are combined for fermentation with Ganoderma lucidum, the total triterpenoid content in the fermentation liquid is higher than that of fermentation with a single strain.

[0135] In Example 6, a combined fermentation of Lactobacillus rhamnosus YYS-B2 and Lactobacillus paracasei YYS-UR produced the most significant increase in the total triterpene content in the Ganoderma lucidum fermentation broth. This suggests that the synergistic effect of YYS-B2 and YYS-UR is more effective than other mixed strains in promoting the release or conversion of bioactive components in Ganoderma lucidum, further increasing the total triterpene content.

[0136] In Example 7, the fermentation broth using a combination of three strains produced a lower total triterpene content than the fermentation broth produced by fermenting with any other two strains. This suggests that when two strains are used to ferment Ganoderma lucidum, a good symbiotic relationship is formed. The substances produced by one strain can be used as substrates by the other strain, promoting the orderly progress of fermentation. However, when three strains are fermented simultaneously, the metabolites produced are more diverse and complex in composition, and some metabolites may inhibit the growth of one or more of the strains and the activity of fermentation-related enzymes.

[0137] Effect of strain ratio:

[0138] As shown in Examples 6-1 to 6-5, when the mass ratio of Lactobacillus rhamnosus YYS-B2 to Lactobacillus paracasei YYS-UR in the mixed strains was 2:3, the total triterpene content in the Ganoderma lucidum fermentation liquid was most significantly improved. The results showed that when the mass ratio of Lactobacillus rhamnosus YYS-B2 to Lactobacillus paracasei YYS-UR in the mixed strains was 2:3, the optimization of the strain synergy, the optimization of the fermentation environment, and the full utilization of nutrients were achieved. These factors work together to significantly improve the total triterpene content in the Ganoderma lucidum fermentation liquid.

[0139] As can be seen from Example 6-1 and Comparative Examples 3 to 4, the mixed strains of Lactobacillus rhamnosus YYS-B2 and Lactobacillus paracasei YYS-UR have a higher rate of improvement in the total triterpene content in the Ganoderma lucidum fermentation liquid than other mixed strains. This indicates that there is a synergistic effect between Lactobacillus rhamnosus YYS-B2 and Lactobacillus paracasei YYS-UR, which enables the two to promote each other during mixed fermentation, thereby increasing the production of total triterpenes. In addition, compared with the commercially available Lactobacillus paracasei F-19 in Comparative Examples 3 and 4, there may be less competition or inhibitory effect between YYS-B2 and YYS-UR, thereby reducing the waste of nutrients and the accumulation of metabolites, and being conducive to the production of total triterpenes.

[0140] Effect of fermentation time:

[0141] As shown in Examples 6, 8, and 10, when the fermentation time of Ganoderma lucidum was 18 to 48 hours, the total triterpene content of the fermentation broth increased most significantly when the fermentation time was 18 hours. However, when the fermentation time was extended to 24 hours, 36 hours, and 48 hours, the total triterpene content in the fermentation broth was lower than that of the fermentation broth at 18 hours. This may be because the growth rate of the mycelium slows down with the extension of the fermentation time, the metabolic activity decreases, the consumption of nutrients and the accumulation of metabolites are accelerated, and the synthesis rate of triterpenoid compounds is reduced.

[0142] Effect of oligofructose addition:

[0143] As shown in Examples 6, 11-13, and Comparative Example 5, the addition of oligofructose ranged from 2% to 5%. In Example 12, the addition of 4% oligofructose significantly increased the total triterpenoid content in the Ganoderma lucidum fermentation broth. This may be because the addition of 4% oligofructose achieved a more optimal microbial community balance, which was more conducive to the synthesis of total triterpenoids in Ganoderma lucidum.

[0144] Result Example 2: Antioxidant Activity of Ganoderma Lucidum Liquid Prepared in Examples and Comparative Examples

[0145] 20 mL of the Ganoderma lucidum liquid of Examples 1 to 3, 6-1 and 12 and the comparative example was taken as a sample liquid, and the DPPH free radical scavenging rate, hydroxyl free radical scavenging rate, ABTS free radical scavenging rate and superoxide anion scavenging rate were tested. The specific data are shown in Table 2.

[0146] Among them, the superoxide anion scavenging rate detection method of Ganoderma lucidum liquid refers to the "Research on the Extraction Process and Antioxidant Activity of Ganoderma Lucidum Leaf Polysaccharide"; the DPPH free radical scavenging rate and ABTS free radical scavenging rate detection methods refer to the DPPH method and ABTS method of antioxidant determination in "GB / T 39100-2020"; the hydroxyl radical scavenging rate detection method refers to the "Research on the Antioxidant Activity of Panax Notoginseng Polysaccharide".

[0147] Table 2 Antioxidant activity of Ganoderma lucidum liquid in Examples and Comparative Examples

[0148]

[0149] As can be seen from Table 2, the free radical scavenging rates of the embodiments of the present invention are all higher than those of the comparative examples, among which the DPPH free radical scavenging rate can reach more than 84%, the hydroxyl free radical scavenging rate can reach more than 20%, the ABTS free radical scavenging rate can reach more than 80%, and the superoxide anion scavenging rate can reach more than 17%.

[0150] It can be seen from Examples 6-1 and 12 that after fermenting Ganoderma lucidum with a combination of Lactobacillus rhamnosus YYS-B2 and Lactobacillus paracasei YYS-UR, the DPPH free radical scavenging rate, hydroxyl free radical scavenging rate, ABTS free radical scavenging rate, and superoxide anion scavenging rate of the Ganoderma lucidum fermentation liquid are all stronger than those of the Ganoderma lucidum fermentation liquid fermented with a single strain.

[0151] Compared with Comparative Example 1, the antioxidant activity of the Ganoderma lucidum in the present invention was improved after fermentation. Antioxidant activity is extremely important for improving immunity. Excessive free radicals can damage the immune system, and scavenging free radicals can protect immune cells.

[0152] In Comparative Example 2, commercially available F-19 Lactobacillus paracasei was used to ferment Ganoderma lucidum under conditions where different strains were used. The free radical scavenging rates of the Ganoderma lucidum fermentation broths in Examples 1 to 3, 6-1, and 12 were all higher than those in Comparative Example 2. This indicates that the strain used in the present invention not only significantly increases the total triterpene content, but also has a better effect on improving the free radical scavenging rate than the commercially available F-19 Lactobacillus paracasei.

[0153] Free radicals are atoms or molecules with unpaired electrons produced by metabolic processes and are highly chemically active. Excessive free radicals in the human body can attack immune cells, disrupt the immune system, and accelerate their aging and death. Eliminating excess free radicals in the body can protect immune cells, maintain the stability of immune system function, and reduce their aging and death. The above results and analysis demonstrate that the Ganoderma lucidum fermented liquid produced by the present invention has a high antioxidant efficacy and can effectively scavenge excess free radicals in the body, thereby enhancing immunity.

[0154] Results Example 3: Effects of Ganoderma lucidum liquid prepared in Examples and Comparative Examples on phagocytic activity and acid phosphatase activity of RAW264.7 cells

[0155] Count the RAW264.7 cells in the logarithmic growth phase and adjust the cell suspension concentration to 5×10 4 / mL, 100μL per well, inoculated into 96-well plates, placed in a 37℃, 5% CO2 incubator for 24h. 20mL of the Ganoderma lucidum liquid of Examples 1 to 3, 6-1 and 13 and the comparative example was taken as a sample liquid, and diluted with 10% FBS DMEM complete medium to a final concentration of 50μL / mL. The culture was continued for 24h, and the effect of the Ganoderma lucidum liquid on cell phagocytosis was detected by the neutral red method, and the acid phosphatase activity was detected by an acid phosphatase kit. The specific data are shown in Table 3. The neutral red method refers to "In vitro antioxidant and immune activity of Juncao Ganoderma lucidum ethanol extract".

[0156] Table 3 Effects of Ganoderma lucidum liquid of Examples and Comparative Examples on phagocytic activity of RAW264.7 cells

[0157] project Cell phagocytosis index (%) Acid phosphatase activity (%) Example 1 104.89±2.70 111.76±3.92 Example 2 108.39±4.74 114.62±0.35 Example 3 109.19±3.50 127.78±1.62 Example 6-1 115.39±1.64 132.92±1.01 Example 12 126.07±3.84 133.57±0.40 Comparative Example 1 102.32±1.00 105.53±2.50 Comparative Example 2 103.02±2.15 109.62±1.78

[0158] When the body is attacked by foreign bacteria or viruses, the human immune system kicks in, and phagocytes begin to identify and engulf pathogens. The level of their phagocytic activity determines the strength of the body's immunity. Acid phosphatase is present in lysosomes within phagocytes and participates in the digestive process within phagocytes, breaking down engulfed material after fusion of the phagosome and lysosome. Acid phosphatase activity is associated with the digestive capacity of phagocytes and participates in intracellular signaling, enabling phagocytes to accurately identify pathogens, transmit signals, and activate immune responses. Enhancing the acid phosphatase activity of phagocytes helps boost immunity. Enhancing the phagocytic capacity and acid phosphatase activity of phagocytes helps them function better, thereby enhancing the body's immunity.

[0159] As shown in Table 3, compared with Comparative Example 1, in Examples 1 to 3 and 6-1, the phagocytic index and acid phosphatase activity of RAW264.7 cells were significantly improved after fermentation of Ganoderma lucidum with different single strains or mixed strains.

[0160] It can be seen from Examples 6-1 and 12 that the phagocytic index and acid phosphatase activity of RAW264.7 cells were the strongest when the combination of Lactobacillus rhamnosus YYS-B2 and Lactobacillus paracasei YYS-UR and 4% oligofructose were added to co-ferment Ganoderma lucidum.

[0161] Therefore, it can be shown that the Ganoderma lucidum fermentation liquid prepared by the present invention can improve the phagocytic activity and acid phosphatase activity of RAW264.7 cells, which is better than the unfermented Ganoderma lucidum liquid in comparative example 1, and the selected strain is better than the commercial strain in comparative example 2.

[0162] Results Example 4: Effects of Ganoderma lucidum liquid prepared in Examples and Comparative Examples on the secretion of NO, IL-6, and TNF-α in RAW264.7 cells

[0163] After counting the Raw264.7 cells in the logarithmic growth phase, the cell suspension concentration was adjusted to 5×10 4 / mL, 100μL per well, inoculated into 96-well plates, placed in a 37°C, 5% CO2 incubator and cultured for 24h. 20mL of the Ganoderma lucidum liquid of Examples 1 to 3, 6-1 and 12 and the comparative example was taken as a sample solution, diluted with 10% FBS DMEM complete medium to a final concentration of 50μL / mL, and added to the culture medium for 24h. The blank group and the model group were added with equal volumes of PBS and 1μg / mL LPS respectively and cultured for 24h. The effects of the test on the secretion of nitric oxide (NO), leukocyte antigen IL-6, and tumor necrosis factor (TNF-α) in RAW264.7 cells were detected using a kit. The specific data are shown in Table 4.

[0164] Table 4 Effects of Ganoderma lucidum liquid of Examples and Comparative Examples on the secretion of NO, IL-6 and TNF-α in RAW264.7 cells

[0165] project NO secretion (μmol / L) IL-6 (μmol / L) TNF-α (μmol / L) Blank group 12.98±0.74 22.38±.2.60 36.63±2.30 LPS group 58.45±1.54 30.95±0.48 55.19±3.13 Example 1 40.58±0.90 33.74±1.61 39.23±0.61 Example 2 46.87±2.65 35.17±0.26 40.12±1.93 Example 3 48.82±0.93 36.84±0.59 41.00±0.92 Example 6-1 49.90±0.85 36.58±0.42 42.14±0.95 Example 12 56.58±2.78 37.94±0.52 42.86±1.95 Comparative Example 1 31.79±0.93 26.98±0.59 36.63±2.30 Comparative Example 2 34.53±0.82 27.43±0.26 37.25±0.96

[0166] RAW264.7 cells can produce NO after being properly stimulated. The secretion of NO can regulate the interaction between immune cells and promote the function of T lymphocytes. The secretion of NO helps to improve the body's immunity.

[0167] IL-6 can activate a variety of immune cells, such as T cells and B cells, thereby initiating an immune response. IL-6 is also an important inflammatory mediator that can induce an inflammatory response, prompting immune cells to aggregate at sites of inflammation and enhancing the body's ability to eliminate pathogens. TNF-α is an important cytokine that can induce or regulate the production of other cytokines. For example, TNF-α immune cells can stimulate immune cells to secrete inflammatory cytokines such as interleukin-6 (IL-6), maintaining the balance of the immune response through interactions with various cytokines. Cytokines such as IL-6 and TNF-α have a synergistic effect, acting together on the immune system to jointly regulate the immune response process, while activating natural killer cells and cytotoxic T cells, enhancing their ability to kill virus-infected cells. Increased secretion of IL-6 and TNF-α indicates that immune cells are activated, thereby improving the body's immunity.

[0168] As shown in Table 4, the secretion levels of NO, IL-6, and TNF-α in RAW264.7 cells after intervention with the Ganoderma lucidum fermentation broth prepared in the examples of the present invention were higher than those after intervention with the Ganoderma lucidum fermentation broth prepared in the comparative example;

[0169] Comparative Example 2 used the commercially available F-19 Lactobacillus paracasei strain. Only under the condition of different strains, after the Ganoderma lucidum fermentation liquid in Examples 1 to 3 was intervened in RAW264.7 cells, the secretion of NO, IL-6, and TNF-α was higher than that of Comparative Example 2;

[0170] It can be seen from Example 12 that, compared with Examples 1 to 3 and Example 6-1, when 4% oligofructose is added and Ganoderma lucidum is fermented with a mixed strain of Lactobacillus rhamnosus YYS-B2 and Lactobacillus paracasei YYS-UR, the production of NO, IL-6, and TNF-α by RAW264.7 cells is significantly increased, indicating that the mixed strain fermentation selected by the present invention not only improves the total triterpene content, antioxidant activity, and cellular phagocytic activity of Ganoderma lucidum, but also improves the secretion of cellular NO, IL-6, and TNF-α due to the commercially available F-19 Lactobacillus paracasei.

[0171] The results show that the Ganoderma lucidum fermentation liquid of the present invention can enhance the phagocytic activity, acid phosphatase activity, NO release, TNF-α release and IL-6 release of mouse RAW264.7 cells, thereby playing a role in improving immunity.

[0172] Results Example 5: Sensory Evaluation of Ganoderma Lucidum Liquid Prepared in Examples and Comparative Examples

[0173] Table 5 Sensory evaluation score sheet

[0174]

[0175] Table 6 Sensory evaluation results of examples and comparative examples

[0176] project Sensory score Example 1 82.33±1.75 Example 2 83.67±0.68 Example 3 83.43±0.99 Example 6 86.89±1.58 Example 12 89.46±0.69 Comparative Example 1 78.42±1.35 Comparative Example 2 81.58±0.85

[0177] Compared with the unfermented comparative example 1, the Ganoderma lucidum extracts in Examples 1 to 3, 6-1 and 12, after fermentation by lactic acid bacteria, have a rich aroma, a mellow taste, a balance of sourness and sweetness, and the sensory scores are all improved.

[0178] Compared to Examples 1 to 3, which used a single strain of Ganoderma lucidum to ferment, Examples 6-1 and 12 used mixed strains to ferment Ganoderma lucidum, resulting in the best sensory scores. This indicates that fermentation with a specific lactic acid bacteria combination significantly improved the sensory scores of the fermented broth.

[0179] In addition, by comparing Example 6-1 and Example 12, it can be seen that the specific lactic acid bacteria combination (YYS-B2 and YYS-UR mixed at a mass ratio of 2:3) used in the present invention, combined with the preferred 4% added fructo-oligosaccharide, can further fully enhance the flavor of the product.

[0180] Comparative Example 2 was compared with the embodiment of the present invention. Both were fermented, and the fermentation flavor and taste were similar, and the sensory score results were not much different.

[0181] It should be noted that:

[0182] (1) Definition:

[0183] As used herein, the term "food" is used in a broad sense to include food and drink for humans. In certain embodiments, the food product is suitable for and designed for human consumption. The Ganoderma lucidum fermentation broth of this application can be used to prepare solid preparations such as powders, tablets, and gels, and can also be dispersed in liquids to prepare liquid preparations, including but not limited to the embodiments.

[0184] "DPPH" in this article is 1,1-diphenyl-2-trinitrophenylhydrazine, also known as 1,1-diphenyl-2-picrylhydrazyl (free radical).

[0185] Herein, “ABTS” refers to the free radical 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid.

[0186] "DMEM" herein refers to Dulbecco's essential minimal medium.

[0187] “PBS” in this article refers to phosphate buffered saline.

[0188] (2) Application of Ganoderma lucidum fermentation liquid:

[0189] Ganoderma lucidum fermented liquid has the following characteristics:

[0190] 1. Increase the content of total triterpenes;

[0191] 2. Possesses antioxidant activity;

[0192] 3. Improve the phagocytic activity of phagocytes;

[0193] 4. Possesses the activity of promoting NO secretion in phagocytes;

[0194] 5. It has the activity of promoting the secretion of TNF-α and IL-6 in phagocytes.

[0195] Based on this, according to the functional characteristics 1-5 above, Ganoderma lucidum fermentation liquid can also be used in functional products with antioxidant, immunity-enhancing and other effects (functional products can be food, health products, cosmetics, skin care products, etc., including any substances that provide preventive and / or other beneficial effects), including but not limited to antioxidant, immunity-enhancing and other effects.

[0196] In summary, the specific parameters or some commonly used reagents or raw materials in the above embodiments are specific embodiments or preferred embodiments under the concept of the present invention, but are not intended to limit the present invention. Those skilled in the art can make adaptive adjustments within the concept and protection scope of the present invention.

[0197] In addition, unless otherwise specified, the raw materials used may also be conventional commercial products in the art, or prepared by conventional methods in the art; that is, the reagents and instruments used in this embodiment are not marked with information such as the manufacturer, and are all conventional products that can be purchased on the market.

[0198] The above description is only a preferred embodiment of the present invention and is not intended to limit the design of this case. Any equivalent changes made based on the key design of this case shall fall within the scope of protection of this case.

Claims

1. A method for preparing a Ganoderma lucidum fermentation liquid for improving total triterpenoid content and immunity, characterized in that: The following steps are involved: Step 1: Pretreatment of raw materials: Ganoderma lucidum is crushed until there are no lumps, and then Ganoderma lucidum powder is mixed with deionized water at a mass ratio of 1:20 to 30 to obtain a Ganoderma lucidum mixed solution; Step 2: Extraction: extracting the Ganoderma lucidum mixture with hot water to obtain Ganoderma lucidum extract; Step 3: Sterilization before fermentation: Heat the Ganoderma lucidum extract to boiling for sterilization, and then cool it down after sterilization; Step 4, fermentation: adding 0.005-0.02% of lactic acid bacteria and 2-5% of oligofructose to the cooled Ganoderma lucidum extract at a temperature of 36-38° C. for 18-48 hours to obtain Ganoderma lucidum fermentation liquid; The added lactic acid bacteria are a mixture of one or more of Lactobacillus plantarum MPs-68, Lactobacillus rhamnosus YYS-B2 and Lactobacillus paracasei YYS-UR; Among them, the deposit number of Lactobacillus plantarum MPs-68 is CGMCC No.31976; the deposit number of Lactobacillus rhamnosus is CGMCC No.28164; the deposit number of Lactobacillus paracasei YYS-UR is CGMCC No.28533; Step 5, solid-liquid separation: performing solid-liquid separation on the Ganoderma lucidum fermentation liquid to obtain Ganoderma lucidum fermentation supernatant; Step 6: Sterilization after fermentation: sterilize the Ganoderma lucidum fermentation supernatant to obtain a sterilized Ganoderma lucidum fermentation liquid.

2. The method for preparing a Ganoderma lucidum fermentation liquid for improving total triterpenoid content and immunity according to claim 1, characterized in that: In the second step of hot water extraction, the Ganoderma lucidum mixture is extracted at 90-100° C. for 160-240 minutes; Step 3: Sterilization: Heat the Ganoderma lucidum extract to boiling and keep it for 20 to 30 minutes for sterilization. After sterilization, cool the Ganoderma lucidum extract to 37°C ± 2°C.

3. The method for preparing a Ganoderma lucidum fermentation liquid for improving total triterpenoid content and immunity according to claim 1, characterized in that: In step 4, during the fermentation process, the lactic acid bacteria strains are composed of any two of Lactobacillus plantarum MPs-68, Lactobacillus rhamnosus YYS-B2, and Lactobacillus paracasei YYS-UR; When the lactic acid bacteria powder consists of Lactobacillus plantarum MPs-68 and Lactobacillus rhamnosus YYS-B2, the mass ratio of the two is 1-3:1-4; When the lactic acid bacteria powder consists of Lactobacillus plantarum MPs-68 and Lactobacillus paracasei YYS-UR, the mass ratio of the two is 1-3:1-5; When the lactic acid bacteria powder consists of Lactobacillus rhamnosus YYS-B2 and Lactobacillus paracasei YYS-UR, the mass ratio of the two is 1-4:1-5; The number of viable bacteria of the Lactobacillus plantarum MPs-68 powder is 1 to 5×10 11 CFU / g; The number of viable bacteria of the Lactobacillus rhamnosus YYS-B2 bacterial powder is 1 to 5×10 11 CFU / g; The viable bacteria count of the Lactobacillus paracasei YYS-UR bacterial powder is 1 to 5×10 11 CFU / g.

4. The method for preparing a Ganoderma lucidum fermentation liquid for improving total triterpenoid content and immunity according to claim 3, characterized in that: During the fermentation process in step 4, the lactic acid bacteria powder is composed of Lactobacillus rhamnosus YYS-B2 and Lactobacillus paracasei YYS-UR, and the mass ratio of the two is 1-3:2-4; Alternatively, the mass ratio of the two is 1.5-2.5:2.5-3.

5.

5. The method for preparing a Ganoderma lucidum fermentation liquid for improving total triterpenoid content and immunity according to claim 1, characterized in that: During the fermentation process in step 4, the lactic acid bacteria strains are composed of Lactobacillus plantarum MPs-68, Lactobacillus rhamnosus YYS-B2 and Lactobacillus paracasei YYS-UR, and the mass ratio of the three is 1-3:1-4:1-5; The number of viable bacteria of the Lactobacillus plantarum MPs-68 powder is 1 to 5×10 11 CFU / g; The number of viable bacteria of the Lactobacillus rhamnosus YYS-B2 bacterial powder is 1 to 5×10 11 CFU / g; The viable bacteria count of the Lactobacillus paracasei YYS-UR bacterial powder is 1 to 5×10 11 CFU / g.

6. The method for preparing a Ganoderma lucidum fermentation liquid for improving total triterpenoid content and immunity according to claim 5, characterized in that: During the fermentation process of step 4, the lactic acid bacteria strains are composed of Lactobacillus plantarum MPs-68, Lactobacillus rhamnosus YYS-B2 and Lactobacillus paracasei YYS-UR, and the mass ratio of the three is 1.5-2.5:1.5-3.5:1.5-4.

5.

7. The method for preparing a Ganoderma lucidum fermentation liquid for improving total triterpenoid content and immunity according to claim 1, characterized in that: The Lactobacillus plantarum MPs-68, Lactobacillus rhamnosus YYS-B2 and Lactobacillus paracasei YYS-UR are provided in the form of bacterial powder or bacterial liquid.

8. The method for preparing a Ganoderma lucidum fermentation liquid for improving total triterpenoid content and immunity according to claim 1, characterized in that: In the solid-liquid separation process of step 5, the Ganoderma lucidum fermentation liquid is first filtered using a 300-mesh sieve, and then centrifuged using a centrifuge to separate the solid and liquid. The centrifugation condition is 6000 r / min for 10 min, and then the Ganoderma lucidum fermentation supernatant is collected; Step 6: Sterilization after fermentation: sterilization conditions are 100-110°C, 20-30 minutes.

9. A Ganoderma lucidum fermentation liquid for improving total triterpenoid content and immunity, characterized by: The preparation method is described in any one of claims 1 to 8.

10. A functional product containing Ganoderma lucidum fermentation liquid, characterized by: Its ingredients include the Ganoderma lucidum fermentation liquid with improved total triterpenoid content and immunity as claimed in claim 9; or its ingredients include the Ganoderma lucidum fermentation liquid with improved total triterpenoid content and immunity prepared by the preparation method according to any one of claims 1 to 8.

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