Aspergillus niger M5.1 and application

Abstract

The invention belongs to the technical field of biology, and particularly relates to aspergillus niger M5.1 and application. The aspergillus niger M5.1 is Aspergillus niger, the preservation number is CGMCC No.40835, the aspergillus niger M5.1 is used for establishing a pseudo-ginseng liquid state fermentation system, a foundation is laid for generating pseudo-ginseng rare saponin through biotransformation of pseudo-ginseng by using the aspergillus niger, and the problems that a method for industrially extracting the pseudo-ginseng rare saponin is poor in water solubility of a product, low in extraction efficiency, incapable of achieving biotransformation of macromolecular saponin, low in extraction cost and the like are solved. The content of the obtained rare saponin is low.

Description

[0001] Parent patent number: 2024111719074, application date: 2024-08-26, patent name: Method for preparing a composition containing rare notoginseng saponin Rh1 using Aspergillus niger M5.1 Technical Field

[0002] The invention belongs to the field of biotechnology, and particularly relates to Aspergillus niger M5.1 and applications thereof. Background Art

[0003] Panax notoginseng (Burk.) FHChen is a plant of the Araliaceae family and the genus Panax Linn. It is a unique health food raw material in China and a unique biological resource in Yunnan Province. It has a long history of medicinal and edible use. Its stems, leaves, flowers (aerial parts), dry roots, and fibrous roots (underground parts) all have high nutritional value. For example, Panax notoginseng fibrous roots are an important raw material for making Yunnan's famous medicinal food Panax notoginseng steamed pot chicken. With the popularization of the concept of big health, modern society attaches more and more importance to the concepts of "health", "green", and "environmental protection". Therefore, the concept of medicine and food homology has become an important goal and policy for the modern application of traditional Chinese medicine.

[0004] At present, the research on the effective active substances of Panax notoginseng has found that the rare saponins of Panax notoginseng have the characteristics of higher biological activity and high expression at low doses. For example, Li Xin, Xu Xu and others pointed out in their research on hemostatic markers of Panax notoginseng that ginsenoside Rh1 is the material basis for Panax notoginseng to contract blood vessels and exert hemostatic effects (Study on the quality markers of Panax notoginseng powder based on hemostatic effects [J]. World Science and Technology-Modernization of Traditional Chinese Medicine, 2022, 24(01): 47-54). In addition, Li Qiuyu screened the hemostatic active ingredient group of Panax notoginseng by the spectrum-effect correlation method. His in vitro hemostatic efficacy experiment pointed out that the hemostatic active ingredients that reduce the plasma recalcification time are mainly: Panax notoginseng (also known as dencichine), Ft1, and Rg2, while F1, Rb1, Re, and Rd have anticoagulant effects (Study on the quality evaluation of Panax notoginseng with different commercial specifications based on hemostatic efficacy [D]. Beijing University of Chinese Medicine, 2021).

[0005] In terms of the biotransformation of rare saponins of Panax notoginseng, Aspergillus can effectively degrade ginsenosides. Huang Chunying, Hou Xiaotao and others fermented Panax notoginseng residue with Aspergillus niger strain alone and found that the polysaccharide content in Panax notoginseng residue increased by 5.03% after fermentation by Aspergillus niger compared with that before fermentation (Changes in the content of polysaccharides in Panax notoginseng residue before and after fermentation by different strains [J]. Journal of Food Safety and Quality, 2020, 11(10): 3164-3168); Yan Bingxiong, Xu Wendi and others found that after Panax notoginseng was fermented by Aspergillus niger, 10 rare saponins including ginsenoside F1, CK and Panax notoginseng saponin Rh16 were detected in its fermentation products (Study on the changes of saponins in Panax notoginseng medicinal materials fermented by Aspergillus niger solid fermentation [J]. China Brewing, 2015, 34(09): 41-44). Zhang Yuanyuan, Yang Qiliang and others explored the effect of Bacillus coagulans on the effective components of Panax notoginseng leaf tea and found that the content of total saponins in Panax notoginseng leaf tea increased significantly by 0.085 to 1.45 times after inoculation with Bacillus coagulans, and was beneficial to improving the antioxidant activity and sensory evaluation of the alcohol extract of the fermentation product (Study on the Antioxidant Activity and Aroma Components of Panax notoginseng Leaf Tea Fermented by Bacillus coagulans [J]. Food and Fermentation Industries, 2023, 49(10): 9-16).

[0006] With the deepening of modern medicine's research on the extraction process, physiological and biochemical properties, pharmacological effects and modern medical applications of Aspergillus niger fermentation products, it was found that Aspergillus niger itself has rich metabolites, including a variety of extracellular polysaccharides, amino acids, amylase, etc. At the same time, its fermentation products have many functions, such as lowering blood sugar and controlling weight, improving immunity, and anti-tumor. On the other hand, Aspergillus niger has a rich and powerful enzyme system, including cellulase, pectinase, amylase, glycoside hydrolase, mannanase, etc., and has great potential for the transformation and modification of active ingredients in medicinal plants. In addition, Aspergillus niger can directly ferment Panax notoginseng medicinal materials to form a two-way fermentation combination.

[0007] Modern industrial methods for extracting rare saponins from Panax notoginseng have problems such as poor water solubility, low extraction efficiency, complex extraction process, low extraction concentration, and other cost-consuming problems. In addition, the extraction of rare saponins from Panax notoginseng using methods such as organic solvent reflux, ultrasound, and extraction cannot achieve the biotransformation of macromolecular saponins, that is, the content of the obtained rare saponins is low. Summary of the invention

[0008] The purpose of the present invention is to provide a method for preparing a composition containing rare notoginseng saponin Rh1 by using Aspergillus niger M5.1, constructing a single substrate bidirectional liquid fermentation mode with Aspergillus niger M5.1 and Panax notoginseng, adopting high concentration for biotransformation, utilizing the rich enzyme system and bacterial metabolic activity of Aspergillus niger M5.1, transforming and modifying the active ingredients of Panax notoginseng, introducing hydroxyl groups, improving the water solubility of rare notoginseng saponins, thereby obtaining a high-concentration Panax notoginseng rare saponin product with good water solubility. In particular, during the fermentation process, no other carbon source and nitrogen source other than Panax notoginseng is added, which effectively reduces the burden of subsequent product separation and purification, reduces the complexity of the fermentation components, and ensures the purity of the rare saponin product.

[0009] On the one hand, the present invention provides an Aspergillus niger M5.1 strain, the strain preservation number is CGMCC No. 40835, the microbial classification is named Aspergillus niger, and it is deposited in the General Microbiology Center of China Microbiological Culture Collection Administration, the preservation address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the preservation time is September 25, 2023.

[0010] Another aspect of the present invention provides a method for preparing a composition containing rare notoginseng saponin Rh1 by using Aspergillus niger M5.1, comprising the following steps:

[0011] Step 1, selecting an activated Aspergillus niger M5.1 strain to prepare Aspergillus niger M5.1 liquid seed solution;

[0012] Step 2, the raw material of Panax notoginseng is crushed and passed through a 100-mesh sieve, the sieved material is mixed evenly with pure water to form a 100 g / L mixture, and the mixture is sterilized in a fermentation tank at 115° C. to 121° C. for 30 min to obtain a Panax notoginseng fermentation medium;

[0013] Step 3, inoculating 5% (v / v) Aspergillus niger M5.1 seed liquid into the Panax notoginseng fermentation medium, introducing sterile air, aeration ratio of 0.5-0.6:1, stirring at 160 rpm, temperature 28-30° C., culturing for 66-88 h, and completing the fermentation;

[0014] Step 4, taking the fermentation broth and concentrating it by centrifugation, membrane filtration, and nanofiltration to obtain a concentrated solution;

[0015] Step 5, drying the concentrated solution to obtain a composition of notoginseng rare saponin Rh1.

[0016] Furthermore, the step 1 comprises:

[0017] The Aspergillus niger M5.1 strain was selected from the culture preservation tube, activated by streaking on a PDA solid culture medium, and cultured at a constant temperature of 30°C for 5 days; the activated Aspergillus niger M5.1 strain seed block was taken and inoculated into a PDA liquid culture medium, and cultured in a shaking flask at 30°C and 160rpm for 3 days to obtain the Aspergillus niger M5.1 seed liquid.

[0018] Furthermore, the step 3 comprises:

[0019] Aspergillus niger M5.1 seed liquid, inoculation amount 2%, culture temperature 30℃, total fermentation time 88h. After inoculation of Aspergillus niger M5.1, air was introduced into the tank with a ventilation ratio of 1:0.45 and a stirring speed of 150rpm for the fermenter. When the dissolved oxygen dropped to 3%, the ventilation ratio was adjusted to 1:0.7 and the speed was increased to 300rpm.

[0020] Furthermore, the step 4 comprises:

[0021] The fermentation broth was centrifuged at 8000 rpm for 10 min, and the supernatant was filtered through a 100,000 Da microporous filter membrane to retain the bacteria and macromolecular compounds to obtain a permeate; the permeate was filtered through a 10,000 Da ultrafiltration membrane to retain small molecular saponins to obtain a retentate, and the retentate was concentrated through a 200 Da nanofiltration membrane to obtain a concentrated solution.

[0022] Furthermore, the step 5 comprises:

[0023] The concentrated liquid is taken and dried by high-temperature forced air to obtain a rare saponin composition of Panax notoginseng, i.e., Panax notoginseng fermented high-temperature dried powder.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] (1) The establishment of a two-way liquid fermentation system with Aspergillus niger M5.1 and Panax notoginseng as a single substrate laid the foundation for Aspergillus niger biotransformation of Panax notoginseng to produce rare Panax notoginseng saponins. The fermentation product with excellent water solubility and rich in rare saponins can be obtained by Aspergillus niger M5.1 biotransformation. The results of HPLC showed that after fermentation, notoginseng saponin R1 and ginsenoside Rg1 were transformed, resulting in increased contents of ginsenoside Rg2, Rh1, Rk1 and notoginseng saponin Ft1. Among them, the growth rate of notoginseng saponin Ft1 reached 834.36%, the growth rate of ginsenoside Rh1 reached 568.86%, the growth rate of ginsenoside Rk1 reached 329.76%, and the growth rate of ginsenoside Rg2 reached 30.83%.

[0026] (2) Verification by LPS-induced mouse macrophage (Raw264.7) inflammation model: At a concentration of 1 μg / mL, both the ginsenoside Rh1 standard and the rare notoginseng saponin combination had the effect of inhibiting the inflammatory factor IL-6 (P < 0.01, the difference was statistically significant). And compared with the ginsenoside Rh1 standard, the rare notoginseng saponin combination prepared by liquid fermentation of Aspergillus niger M5.1 had a significantly higher inhibitory effect on the inflammatory factor IL-6 than the ginsenoside Rh1 standard (P < 0.01, the difference was statistically significant). BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 It is a flow chart of a method for preparing a composition containing rare notoginseng saponin Rh1 by using Aspergillus niger M5.1 of the present invention;

[0028] Figure 2 is the growth rate of Rh1 saponin relative to Panax notoginseng raw material during the fermentation process in one embodiment of the present invention;

[0029] Figure 3 is the ratio of the content of Rh1 saponin to the total saponin content of Panax notoginseng in the fermentation broth during the fermentation process in one embodiment of the present invention;

[0030] Figure 4 is the change of dissolved oxygen, pH and biomass during the fermentation process in one embodiment of the present invention;

[0031] Figure 5 is the cell viability after stimulation with different samples at a concentration of 1 μg / mL in one embodiment of the present invention;

[0032] Figure 6 The effect of the saponin standard and the rare saponin composition sample of Panax notoginseng on the concentration of inflammatory factor IL-6 in one embodiment of the present invention;

[0033] Figure 7 It is the inhibition rate of the saponin standard and the Panax notoginseng rare saponin composition sample on the inflammatory factor IL-6 in one embodiment of the present invention;

[0034] Figure 8 is the skin lesion healing rate of SD rats in one embodiment of the present invention;

[0035] Fig. 9 1 is a diagram showing the healing of skin lesions on SD rats in one embodiment of the present invention; a is a diagram showing the healing of skin lesions on SD rats before administration, and b is a diagram showing the healing of skin lesions on SD rats after application of the rare saponin composition of Panax notoginseng. DETAILED DESCRIPTION

[0036] The present invention is described in detail below in conjunction with the various embodiments shown in the accompanying drawings, but it should be noted that these embodiments are not limitations of the present invention, and any equivalent transformations or substitutions in functions, methods, or structures made by ordinary technicians in the field based on these embodiments are all within the scope of protection of the present invention.

[0037] Ginseng Figure 1 As shown, this embodiment provides a method for preparing a composition containing rare saponin Rh1 of Panax notoginseng using Aspergillus niger M5.1, which is mainly composed of rare saponin Rh1 of Panax notoginseng by fermenting Panax notoginseng using Aspergillus niger M5.1, and its contents mainly include three parts: fermentation and conversion of Panax notoginseng culture medium, molecular sieve separation of rare saponins of Panax notoginseng, and preparation of high-temperature drying powder of the composition. The Aspergillus niger M5.1 strain used in this embodiment has been registered and preserved in the General Microbiological Center of the China Microbiological Culture Collection Administration, and the strain preservation number is CGMCC No.40835. The microorganism classification is named Aspergillus niger, and it is preserved in the General Microbiological Center of the China Microbiological Culture Collection Administration, and the preservation address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the preservation time is September 25, 2023. Aspergillus niger M5.1 strain, wherein M5.1 is a self-numbered number.

[0038] 1. Fermentation of Panax notoginseng fermentation medium

[0039] 1) Sterilization of culture medium

[0040] The raw materials of Panax notoginseng were crushed and passed through a 100-mesh sieve and mixed evenly with pure water. The total liquid volume of the fermentation tank was 30 L. After sterilization at 121°C for 30 minutes, the Panax notoginseng fermentation medium was obtained.

[0041] 2) Seed solution preparation

[0042] According to the aseptic operation requirements, the Aspergillus niger M5.1 strain was selected from the strain preservation tube, activated by streaking on PDA solid medium, and cultured at 30°C for 5 days. The strain was observed to be in good growth state without contamination. The activated Aspergillus niger M5.1 strain (cut into a seed block of about 1×1 cm in size) was inoculated into PDA liquid medium, cultured in a shake flask at 30°C, 160rpm for 3 days, and the Aspergillus niger M5.1 seed liquid was obtained.

[0043] 3) Vaccination

[0044] According to the requirements of sterile operation, the Aspergillus niger M5.1 seed liquid was inoculated into the Panax notoginseng fermentation medium, with an inoculation amount of 2%, a culture temperature of 30°C, a total fermentation time of 88h, an air ventilation ratio of 1:0.45, and a rotation speed of 150rpm; when the dissolved oxygen dropped to 3%, the ventilation ratio was adjusted to 1:0.7 and the rotation speed was increased to 300rpm.

[0045] 4) Analysis of fermentation broth components

[0046] Determination of total saponins of Panax notoginseng: Vanillin-glacial acetic acid was used for determination.

[0047] Determination of rare saponins in Panax notoginseng: Determination was carried out by high performance liquid chromatography, with an injection volume of 10 μL; column oven temperature of 30°C; flow rate of 1.0 mL / min; and detection wavelength of 203 nm.

[0048] After 88h of fermentation by Aspergillus niger M5.1, the conversion of rare saponin Rh1 reached its peak. During the fermentation process, the content of Rg1 saponin and R1 saponin decreased, and they were mainly converted into Rh1, Rk1 and Ft1. The changes in the content of saponins after conversion relative to that of the raw material of Panax notoginseng were mainly concentrated in notoginseng saponin Ft1 (the growth rate reached 834.36%), ginsenoside Rh1 (the growth rate reached 568.86%) and ginsenoside Rk1 (the growth rate reached 329.76%) saponins.

[0049] Table 1 Growth rate of rare saponins of Panax notoginseng after fermentation

[0050]

[0051]

[0052] After liquid fermentation, the proportion of rare saponins Ft1, Rh1 and Rk1 in the total saponin content of Panax notoginseng increased. During the fermentation process, R1 and Rg1 decreased significantly.

[0053] Depend on Figure 2 It can be seen that compared with the rare saponins in Panax notoginseng raw materials after fermentation for 64 hours, the content of ginsenoside Rh1 increased by 472.84%.

[0054] 5) Fermentation parameter analysis

[0055] A 50L fermentation tank was used to directly ferment Panax notoginseng powder with Aspergillus niger M5.1. Since the carbon and nitrogen sources used for the growth of Aspergillus niger are all from Panax notoginseng raw materials, the dissolved oxygen, pH and biomass growth during the fermentation process can be used as guiding parameters for saponin conversion. Figure 4 It can be seen that the process of Aspergillus niger M5.1 fermenting Panax notoginseng is an acid-producing process, and a large amount of Panax notoginseng acid is produced. As the fermentation time extends, the pH of the fermentation liquid shows an acidic change, and the pH is as low as 2.5, indicating that a large amount of organic acids are produced in the system. At the same time, combined with the changes in dissolved oxygen content in the fermentation environment, it can be seen that Aspergillus niger M5.1 is in a logarithmic growth period within 0 to 20 hours after inoculation, the dissolved oxygen value in the tank is at a low value, and the mycelium in the tank grows a lot, accompanied by great oxygen consumption, and the biomass reaches the maximum value at 60 hours, that is, the process of Aspergillus niger M5.1 fermenting Panax notoginseng and biotransforming Panax notoginseng rare saponins needs to be carried out under aerobic conditions.

[0056] Combination Figure 3 As shown, at 88h of fermentation, the content of ginsenoside Rh1 (accounting for 9.4% of the total saponin content of Panax notoginseng in the fermentation broth) reached the maximum conversion value.

[0057] After fermentation by Aspergillus niger M5.1, the content of notoginseng saponin Ft1 (increase rate reached 834.36%), ginsenoside Rh1 (increase rate reached 568.86%) and ginsenoside Rk1 (increase rate reached 329.76%) increased compared with the raw material of Panax notoginseng. After liquid fermentation, the proportion of Rh1, Ft1 and Rk1 saponins increased.

[0058] During the fermentation process, notoginsenoside R1 and ginsenoside Rg1 are consumed and converted into ginsenoside Rh1, ginsenoside Rg2, notoginsenoside Ft1 and ginsenoside Rk1, among which the main concentration is the conversion of notoginsenoside Ft1, ginsenoside Rk1 and ginsenoside Rh1.

[0059] 2. Inhibitory effect of rare notoginseng saponin combination on inflammatory factor IL-6

[0060] Cell plating → sample addition → MTT assay to determine cell viability → cell plating → collection of cell supernatant → MTT test on cells without supernatant → ELISA test → calculation of inflammatory factor inhibition rate.

[0061] 1) Sample maximum safe concentration screening results

[0062] like Figure 5 As shown, the cell viability of ginsenoside Rg1 standard, ginsenoside Rh1 standard, ginsenoside Rb1 standard, ginsenoside F1 standard and notoginsenoside Ft1 standard were all >90% within the concentration range of ≤1 μg / mL, and appropriate concentrations could be selected for determination within the concentration range of ≤1 μg / mL.

[0063] The cell viability of the rare notoginseng saponin composition (Fermented high-temperature dried powder of Panax notoginseng) is >90% within the concentration range of ≤1μg / mL, and the appropriate concentration can be selected for determination within the concentration range of ≤1μg / mL; the content of ginsenoside Rg2 is 1.334mg / g, the content of ginsenoside Rh1 is 1.646mg / g, the content of ginsenoside F1 is 0.297mg / g, the content of notoginseng saponin Ft1 is 2.867mg / g, and the content of ginsenoside Rk1 is 0.333mg / g. In order to compare the effects of saponin standards and the rare notoginseng saponin composition on the inflammatory factor IL-6 at the same concentration, the 1μg / mL concentration was selected for ELISA test.

[0064] 2) Effect of rare notoginseng saponin combination on inflammatory factor IL-6

[0065] like Figure 6 As shown, compared with the blank control group (BC), the IL-6 concentration was significantly increased after LPS stimulation (NC) (***P<0.001); compared with the negative control group (NC), dexamethasone (PC, 100μg / mL) could significantly inhibit the secretion of IL-6 (##P<0.01), indicating that the stimulation conditions were effective.

[0066] like Figure 6 As shown, compared with the negative control group (NC), ginsenoside Rg1 standard, ginsenoside Rh1 standard, notoginseng saponin Ft1 standard and notoginseng rare saponin combination samples can reduce the concentration of inflammatory factor IL-6 at a concentration of 1 μg / mL (the difference is statistically significant). Among them, ginsenoside Rh1 standard and notoginseng rare saponin combination have a more advantageous effect on reducing the concentration of inflammatory factor IL-6 than other saponins.

[0067] like Figure 7 As shown, the inhibition rate of inflammatory factor IL-6 of the rare saponin composition of Panax notoginseng and the saponin standard product showed that at a concentration of 1 μg / mL, the inhibition rate of the rare saponin composition of Panax notoginseng on inflammatory factor IL-6 was significantly higher than that of ginsenoside Rg1 standard product, ginsenoside F1 standard product, ginsenoside Rb1 standard product and Panax notoginseng saponin Ft1 standard product (P < 0.001, the difference was statistically significant). In addition, the inhibitory effect of the rare saponin composition of Panax notoginseng on inflammatory factor IL-6 was significantly higher than that of ginsenoside Rh1 standard product (P < 0.01, the difference was statistically significant).

[0068] 3. Repairing effect of rare notoginseng saponin combination on skin lesions in SD rats

[0069] like Figure 8 As shown, 6 days after applying the rare saponin composition of Panax notoginseng, the healing rate of the skin lesions of SD rats was 44.68%; 6 days after applying normal saline, the healing rate of the skin lesions of SD rats was 38.75%; 6 days after applying penicillin, the healing rate of the skin lesions of SD rats was 37.20%.

[0070] like Fig. 9 As shown, 12 days after applying the rare saponin composition of Panax notoginseng, the wounds on the skin lesions of SD rats were dry and healed well, with a healing rate of 92.57%.

[0071] The present invention utilizes the screened Aspergillus niger M5.1 strain, with strain collection number GMCC No. 40835; the strain directly ferments the raw material of Panax notoginseng without adding any carbon and nitrogen raw materials to the culture medium; the strain can ferment at a concentration of 100g / L-180g / L of Panax notoginseng, and biotransform to generate a composition of rare saponin Rh1. The composition has the efficacy of inhibiting the inflammatory factor IL-6, and has obvious anti-inflammatory effect in animal injury repair experiments.

[0072] The present invention constructs a two-way liquid fermentation system of Aspergillus niger M5.1 strain and Panax notoginseng single substrate, uses HPLC technology to analyze the differences and changes in saponin components before and after fermentation; evaluates the in vitro cell scratch repair activity of Panax notoginseng before and after fermentation, and the prepared Panax notoginseng composition has a repair effect. The specific technical effects are as follows:

[0073] (1) The establishment of a two-way liquid fermentation system with Aspergillus niger and Panax notoginseng as a single substrate laid the foundation for Aspergillus niger biotransformation of Panax notoginseng to produce rare Panax notoginseng saponins. The fermentation product with excellent water solubility and rich in rare saponins can be obtained by Aspergillus niger biotransformation. The results of HPLC showed that after fermentation, notoginseng saponin R1 and ginsenoside Rg1 were transformed, resulting in increased contents of ginsenoside Rg2, Rh1, Rk1 and notoginseng saponin Ft1. Among them, the growth rate of notoginseng saponin Ft1 reached 834.36%, the growth rate of ginsenoside Rh1 reached 568.86%, the growth rate of ginsenoside Rk1 reached 329.76%, and the growth rate of ginsenoside Rg2 reached 30.83%.

[0074] (2) Verification by LPS-induced mouse macrophage (Raw264.7) inflammation model: At a concentration of 1 μg / mL, both the ginsenoside Rh1 standard and the rare notoginseng saponin combination had the effect of inhibiting the inflammatory factor IL-6 (P < 0.01, the difference was statistically significant). In addition, the rare notoginseng saponin combination prepared by liquid fermentation of Aspergillus niger M5.1 had a significantly higher inhibitory effect on the inflammatory factor IL-6 than the ginsenoside Rh1 standard (P < 0.01, the difference was statistically significant).

[0075] The present invention utilizes the screened Aspergillus niger M5.1 strain, with strain collection number GMCC No. 40835; the strain directly ferments the raw material of Panax notoginseng without adding any carbon and nitrogen raw materials to the culture medium; the strain can ferment at a concentration of 100g / L-180g / L of Panax notoginseng, and biotransform to generate a composition of rare saponin Rh1. The composition has the efficacy of inhibiting the inflammatory factor IL-6, and has obvious anti-inflammatory effect in animal injury repair experiments.

[0076] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above and that the present invention can be implemented in other specific forms without departing from the spirit or essential features of the present invention. Therefore, the embodiments should be considered exemplary and non-restrictive in all respects, and the scope of the present invention is defined by the appended claims rather than the above description, and it is intended that all changes falling within the meaning and scope of the equivalent elements of the claims be included in the present invention.

Claims

1. An Aspergillus niger M5.1 strain, characterized in that The strain preservation number is CGMCC No.40835, the microbial classification name is Aspergillus niger, and it is preserved in the General Microbiology Center of China Microorganism Culture Collection Administration. The preservation address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the preservation time is September 25, 2023.

2. Use of the Aspergillus niger M5.1 strain as described in claim 1 in increasing the content of rare saponin Rh1 in Panax notoginseng.

3. Use of the Aspergillus niger M5.1 strain as described in claim 1 in increasing the content of rare saponin Rg2 in Panax notoginseng.

4. Use of the Aspergillus niger M5.1 strain as claimed in claim 1 in increasing the content of rare saponin Rk1 in Panax notoginseng.

5. Use of the Aspergillus niger M5.1 strain as claimed in claim 1 in increasing the content of notoginseng saponin Ft1 in Panax notoginseng.

6. The use according to any one of claims 2 to 5, characterized in that: The fermentation concentration of Panax notoginseng is 100g / L-180g / L.

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