CN117801961A - Trametes versicolor and conversion of ginsenoside Rb 1 Preparation of ginsenoside 20- (S) -Rh 2 、20-(R)-Rh 2 Is a method of (2) - Google Patents
Trametes versicolor and conversion of ginsenoside Rb 1 Preparation of ginsenoside 20- (S) -Rh 2 、20-(R)-Rh 2 Is a method of (2) Download PDFInfo
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Abstract
The invention belongs to the technical field of microorganism application, and in particular relates to a method for converting ginsenoside Rb by using trametes versicolor 1 Preparation of ginsenoside 20- (S) -Rh 2 、20‑(R)‑Rh 2 Is a method of (2). Ginsenoside Rb as main ingredient in Notoginseng radix total saponin by trametes versicolor 1 Performing bioconversion to obtain two ginsenoside 20- (S) -Rh which are isomers 2 、20‑(R)‑Rh 2 The two ginsenosides have various pharmacological activities, and are particularly remarkable in the reverse side of the anti-tumor activity. The conversion method overcomes the shortages of environmental pollution, low conversion efficiency, etc. of the traditional method, and increases ginsenoside 20- (S) -Rh 2 、20‑(R)‑Rh 2 Green high efficiency conversionA new method.
Description
Technical Field
The invention belongs to the technical field of microorganism application, and in particular relates to trametes versicolor and conversion of ginsenoside Rb thereof 1 Preparation of ginsenoside 20- (S) -Rh 2 、20-(R)-Rh 2 Is a method of (2)
Background
Notoginseng radix (Panax notoginseng) is one of traditional Chinese medicines in China, belongs to plants of Panax genus of Araliaceae, is named as radix Stephaniae Sinicae, notoginseng radix, etc., has wide distribution area, and is a traditional rare Chinese herbal medicine in China. Panax Notoginsengs (PNS) is the main active ingredient of Notoginseng radix, and has antiinflammatory, blood lipid reducing, antioxidant and antitumor effects. The main component of Notoginseng radix total saponin comprises Notoginseng radix saponin R 1 And ginsenoside Rb 1 、Rd、Re、Rg 1 Generally, the radix notoginseng root can be used as a medicine and a food for human body. However, the main saponins in the total saponins of the pseudo-ginseng are large polar saponins, have more glucosyl groups, and are difficult to be absorbed in human bodies to exert maximum effect; thus, there is a need to find a method for converting a large polar saponin into a small limit saponin, such as bioconversion in the present method.
Coriolus versicolor (l.) fungus, chinese name: coriolus versicolor; alias name: coriolus versicolor, huang Yunzhi, coriolus versicolor, etc. Wild on various broad-leaved tree piles, inversed trees or branches, and distributed in forests around the world. It is often used as a medicine for its fruiting body, and has the effects of invigorating spleen, promoting diuresis, and clearing heat and detoxicating. However, the application of the coriolus versicolor to the conversion of saponins has not been disclosed yet, and the coriolus versicolor has natural advantages as edible fungi for the conversion of saponins. In addition, the secreted extracellular enzyme also has the function of hydrolyzing glucosyl groups and is more efficient.
Modern pharmacological research proves that ginsenoside 20- (S) -Rh 2 、20-(R)-Rh 2 Has strong physiological activity, and has antitumor, myocardial ischemia improving, endometriosis resisting, antiallergic, immunity regulating, radioprotective, asthma resisting, antidepressant, and pharmacological activities, so that ginsenoside 20- (S) -Rh can be used for treating and preventing cancer 2 、20-(R)-Rh 2 Has extremely high medicinal value and application prospect. The research and development of the preparation method has important significance for discovering components with stronger activity and developing new antitumor drugs. Methods for converting a large polar saponin into a ginsenoside are known as chemical hydrolysis, enzymatic hydrolysis, microbial fermentation, heating processing, chemical synthesis, and the like. However, the existing transformation methods have more or less defects, such as environmental pollution, low transformation rate, high transformation difficulty and the like. Therefore, it is important to develop a method for converting a large polar saponin into a small limit saponin by bioconversion.
Disclosure of Invention
In view of the above, the present invention aims to provide a trametes versicolor strain and its conversion of ginsenoside Rb 1 Preparation of ginsenoside 20- (S) -Rh 2 、20-(R)-Rh 2 The method comprises converting main large polar saponin in Notoginseng radix into ginsenoside 20- (S) -Rh with high physiological activity and easy absorption and utilization by human body by microorganism conversion 2 、20-(R)-Rh 2 Overcomes the shortages of environmental pollution, low conversion efficiency and the like of the traditional method, and adds a new green and high-efficiency conversion method of the saponin.
Specifically, the invention provides a strain of coriolus versicolor, which is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.40999.
The invention also provides a method for converting ginsenoside Rb by using trametes versicolor 1 Preparation of ginsenoside 20- (S) -Rh 2 、20-(R)-Rh 2 The method of (1) uses trametes versicolor to treat ginsenoside Rb 1 Fermenting, extracting to obtain total crude extract, and separating and purifying to obtain ginsenoside 20- (S) -Rh 2 、20-(R)-Rh 2 。
In some embodiments of the invention, thePreparation of ginsenoside 20- (S) -Rh by converting trametes versicolor into ginsenoside Rb1 2 、20-(R)-Rh 2 Is characterized by comprising the following steps:
(1) Culture of the strain: inoculating Coriolus versicolor trametes in culture medium, shake culturing T 1 d；
(2) Ginsenoside Rb 1 Is converted into: after the step (1) of shaking culture, adding substrate ginsenoside Rb 1 Then shake culture T is carried out 2 d, obtaining strain fermentation liquor;
(3) Extracting fermentation liquid: extracting the strain fermentation liquor by adopting an organic solvent, and concentrating the extracting solution under reduced pressure to obtain a total crude extract;
(4) And (3) separating and purifying: the total crude extract is separated and purified to obtain ginsenoside 20- (S) -Rh 2 、20-(R)-Rh 2 。
In some embodiments of the invention, the strain culturing in step (1) above comprises: picking coriolus versicolor keyhole hypha beside an alcohol lamp of an ultra-clean workbench, inoculating into a culture medium, placing in a shaking table for culturing for 3-5 days, sucking coriolus versicolor keyhole bacterial suspension by a liquid-transferring gun, inoculating into the culture medium, and culturing at room temperature for 3-5 days.
In some embodiments of the invention, the medium in step (1) above is PDB medium.
In some embodiments of the invention, the PDB medium is formulated as follows: 5g/L of potato leaching powder; glucose 15g/L is added into deionized water to prepare a culture medium, and sterilization conditions are as follows: pressure 101kpa: sterilizing in a steam autoclave at 115 deg.C for 30min.
In some embodiments of the present invention, the shaking culture speed in the step (1) is 130-150 rmp/min, the culture temperature is 20-30 ℃, and T is not less than 3 ≡T 1 ≤5d。
In some embodiments of the invention, the medium in step (1) above is PDB medium; the shake cultivation speed in the step (1) is 130-150 rmp/min, the cultivation temperature is 20-30 ℃, and T is not less than 3% 1 ≤5d。
In some embodiments of the inventionIn the scheme, the ginsenoside Rb in the step (2) is 1 The preparation steps of (a) are as follows: pulverizing Notoginseng radix main root, extracting with 50% -70% ethanol under reflux to obtain extractive solution, and purifying to obtain ginsenoside Rb 1 。
In some embodiments of the present invention, the ginsenoside Rb in the step (2) above 1 The preparation steps of (a) are as follows: pulverizing Notoginseng radix main root, extracting with 50% ethanol under reflux to obtain extractive solution, and purifying to obtain ginsenoside Rb 1 。
In some embodiments of the present invention, the shaking culture speed in the step (2) is 130-150 rmp/min, the culture temperature is 20-30 ℃, and T is 15-T 2 ≤20d。
In some embodiments of the present invention, the ginsenoside Rb added in the step (2) above 1 The concentration is 0.01-0.1mg/mL.
In some embodiments of the present invention, the ginsenoside Rb added in the step (2) above 1 The concentration was 0.01mg/mL.
In some embodiments of the present invention, the specific operation method of step (2) above comprises: ginsenoside Rb 1 Dissolving in 75% ethanol solution, and adding into culture solution by passing through 0.45 μl sterile filter membrane at ultra-clean bench alcohol lamp to obtain saponin content of 0.01mg/ml; continuing to shake culture for 15 days, and performing ginsenoside Rb 1 To obtain fermentation liquor.
In some embodiments of the present invention, the organic solvent extraction step in step (3) above is as follows: adding water saturated n-butanol with 1-2 times of the liquid volume of the bacteria, centrifuging, retaining n-butanol layer extract, extracting for 3-5 times, mixing the extracts, and concentrating the extract under reduced pressure to obtain total crude extract.
In some embodiments of the present invention, the organic solvent extraction step in step (3) above is as follows: adding water saturated n-butanol with the volume of 1-2 times of the liquid, and centrifuging at 13000rmp/min for 10 min; the n-butanol layer extract is reserved, the extraction is carried out for 3 to 5 times, the extracts are combined, and the total crude extract is obtained by decompressing and concentrating the extracts.
In some embodiments of the present invention, the separation and purification in the step (4) above comprises the steps of: subjecting the total crude extract to final separation and purification by silica gel column chromatography or semi-preparative high performance liquid chromatography or ODS to obtain 20- (S) -Rh 2 And 20- (R) -Rh 2 。
The invention also provides a fermentation liquor obtained by fermenting the trametes versicolor.
The invention also provides the transformation of the trametes versicolor into the ginsenoside Rb 1 Preparation of ginsenoside 20- (S) -Rh 2 And ginsenoside 20- (R) -Rh 2 Is used in the field of applications.
Compared with the prior art, the preparation method of the invention uses microbial transformation to transform the main large polar saponin in the notoginseng root into the ginsenoside 20- (S) -Rh which is easy to be absorbed and utilized by human body and has high physiological activity 2 、20-(R)-Rh 2 The method overcomes the shortages of environmental pollution, low conversion efficiency and the like of the traditional method, adds a new green and efficient conversion method of the saponin, has the advantages of simple conversion method, stable conversion effect, rich conversion products, environmental friendliness and the like, improves the bioavailability of the active ingredients in the pseudo-ginseng, and is suitable for commercial production and preparation.
Drawings
In order to more clearly illustrate the embodiments of the invention or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described, it being obvious that the drawings in the following description are only some embodiments of the invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 shows the morphology of the strain of trametes versicolor isolated and screened in example 1: strain morphology on PDA medium.
FIG. 2 shows the morphology of the strain of trametes versicolor isolated and screened in example 1: the morphology was observed microscopically.
FIG. 3 shows the morphology of the strain of trametes versicolor isolated and screened in example 1: strain morphology on PDB medium.
FIG. 4 is a phylogenetic tree of trametes versicolor isolated and screened in example 1.
FIG. 5 shows the results of thin layer chromatography using trametes versicolor fermentation product of example 3.
FIG. 6 shows the results of a high performance liquid chromatography using trametes versicolor fermentation product of example 3;
FIG. 7 is a schematic flow chart of the preparation and analysis of example 3.
Biological material preservation information: the classification is named as trametes versicolor (Latin academy: trametescescolor), and is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.40999, address: the collection date of the institute of microbiology of the national academy of sciences of China is 2023, 11 months and 27 days.
Detailed Description
The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it should be apparent that the described embodiments are only some of the embodiments of the present invention and should not be used to limit the protection scope of the present invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the present invention without making any inventive effort, are intended to fall within the scope of the present invention.
Example 1: isolation and screening of trametes versicolor (trametes versicolor) strains
Fresh gastrodia elata is taken as a raw material (the gastrodia elata is purchased in the market), soil on the surface of the gastrodia elata is cleaned, and surface sterilization and disinfection (ultra-clean workbench) are carried out by using 75% ethanol and other solutions. After disinfection, the internal tissue of the gastrodia elata is cut into small cubes (1 cm multiplied by 1 cm) by a knife, a plurality of tissue blocks are laid flat and put into a PDA solid culture medium, and the tissue blocks are put into an incubator (25 ℃) for culture after being sealed by a sealing film. And (3) periodically observing the growth condition of the bacterial colonies during the culture period, if bacteria grow out, picking up the bacterial colonies with different forms by using an inoculating needle beside the ultra-clean bench alcohol lamp in time, inoculating the bacterial colonies into a new PDA culture medium, and placing the culture medium into a 25 ℃ incubator for culture. Avoiding the newly grown dominant bacteria from extruding the bacterial colony grown later, and completing the bacterial colony as much as possible in 3-5 days. And repeatedly separating and screening until only single colonies appear in the culture dish, and inoculating the single colonies into a slant test tube, and storing in a refrigerator at 4 ℃.
Example 2: morphological and molecular characterization of strains
The preparation method of the PDA solid medium used in the embodiment comprises the following steps: adding RO water to prepare culture medium according to formula of PDA culture medium (potato extract powder 5g/L, glucose 15g/L, agar 25 g/L), sterilizing at 115 deg.C for 30min, oven drying at 55deg.C, sterilizing in ultra clean bench for 30min, pouring about 25ml culture medium into each culture dish, and cooling.
Morphological and molecular characterization of the strains isolated and screened in example 1:
colony morphology identification: a small amount of mycelium is picked and inoculated on 3 points of PDA solid culture medium symmetrically, after 5d of culture at 25 ℃, the colony morphology as shown in figure 1 is observed, micro fluff is observed at the initial stage of the colony from the colony morphology, the colony is milky white and uniform, the surface is smooth, the mycelium is compact, the growth vigor is strong, the mycelium is thick white, the colony grows radially to the periphery, and no obvious concentric ring belt exists.
And (5) microscopic morphological identification: and (3) selecting mycelia well growing on a PDA solid culture medium plate for tabletting, and observing under an optical microscope to obtain the conidiophore form shown in figure 2, wherein the conidiophore of the strain is single-round or double-round broom-shaped, and the conidiophore is cylindrical.
Colony morphology identification: the mycelium growing well on the PDA solid culture medium plate is picked up and put in PDB liquid culture medium, after 3-5 days of culture, the strain form can be observed, the color is light yellow, the shape of the mycelium is small sphere, and the mycelium is covered with fine fluff, as shown in figure 3.
Sequencing and identifying fungus ITS: the strain is sent to Beijing engine biotechnology limited company for ITS amplification sequencing, and finally the measurement result is submitted to NCBI database for BLAST homology comparison, and the strain of similar species is selected to construct a strain phylogenetic tree shown in figure 4 by using a Neighbor-joining method (Neighbor-joining) in software MEGA11, and the strain can be judged to conform to the characteristics of trametes of Polyporaceae from the phylogenetic tree.
From all the above data, it was confirmed that the strain was trametes versicolor (trametes versicolor) of trametes genus of Polyporaceae family.
Example 3: conversion of ginsenoside Rb by trametes versicolor 1 Preparation of ginsenoside 20- (S) -Rh 2 、20-(R)-Rh 2 Comparative experiments of (2)
(1) PDA solid culture medium preparation
Adding RO water to prepare culture medium according to formula of PDA culture medium (potato extract powder 5g/L, glucose 15g/L, agar 25 g/L), sterilizing at 115 deg.C for 30min, oven drying at 55deg.C, sterilizing in ultra clean bench for 30min, pouring about 25ml culture medium into each culture dish, and cooling.
(2) Expansion culture
In an ultra-clean workbench, a small amount of mycelia are selected from a PDA solid culture medium of a strain with the preservation number of CGMCC No.40999 and cultured in a PDB liquid culture medium for 5d, 1mL of bacterial suspension is absorbed, the bacterial suspension is inoculated into a new PDB liquid culture medium, and the bacterial suspension is cultured for 5d under the condition of room temperature, so that the bacterial is accumulated to a certain extent, and the inhibition of early growth of the strain after the saponin is added is resisted.
Inoculating the seed solution in the sterilized 30-35 LPDB culture medium for expansion culture, and culturing at 20-30 ℃ for 3-5 d in a 150rmp/min shaking table.
The proportion of each component in the PDB liquid culture medium is 5g/L of potato extract powder, 15g/L of glucose, natural pH, 101kPa of pressure and high-temperature high-pressure sterilization at 115 ℃ for 30min.
(3) Conversion of ginsenoside Rb by trametes versicolor 1 Comparative experiments of (2)
Three are arrangedGinsenoside Rb for each group 1 The conversion activity was studied by weighing ginsenoside Rb in the strain control group (no saponin in bacteria), the saponin control group (no saponin in bacteria), and the saponin conversion group (thallus+saponin) 1 Dissolving with 75% ethanol by mass fraction, sucking the solution in an ultra-clean workbench with a sterile syringe, connecting with 0.45 μm sterile filter, and adding ginsenoside Rb into the saponin conversion group and saponin control group 1 The liquid culture medium contains 0.01mg ginsenoside Rb per 1mL 1 The strain is assembled and inoculated with the bacterial suspension in the step (3) without adding ginsenoside Rb 1 The saponin conversion group was inoculated with the bacterial suspension in step (3), and the three groups were further cultured for 15d.
(4) Thin Layer Chromatography (TLC) analysis of the conversion product
The bacterial liquid is taken out and placed in a centrifuge tube, the equal volume of water saturated n-butanol is added for vortex oscillation for 2min, the saponin compounds in the fermentation liquid are fully extracted, centrifugation is carried out for 3min at the rotating speed of 5000r/min, bacterial sedimentation and solution layering are promoted, the supernatant liquid is sucked by a capillary tube, thin-layer chromatography analysis is carried out, the developing agent is trichloromethane-methanol-water-glacial acetic acid (V: V, 6.75:2.5:0.5:0.25), the color reagent is 10% sulfuric acid-ethanol, the TLC analysis result is shown in figure 5, compared with a saponin control group and a strain control group, a plurality of new color development points appear in the saponin conversion group and the positions are higher than that of a substrate, the substances have smaller polarities, so that a larger migration distance is displayed on the thin-layer chromatography plate, and the result indicates that ginsenoside is generated.
(5) High Performance Liquid Chromatography (HPLC) analysis of the conversion products
Taking out the n-butanol upper layer of the extract in the step (4), spin-drying at 55 ℃, dissolving with 1mL chromatographic grade methanol, passing through a 0.45 μm organic system filter membrane, and then analyzing with a high performance liquid chromatograph under the condition of C18 column (5 μm,250mm×4.6 mm); the detection wavelength is 203nm; column temperature is 30 ℃; volume flow rate is 1.0mL/min; the sample injection amount is 20 mu L; mobile phase aqueous solution (a) -acetonitrile (B); the elution procedure is 0-25 min,20% B; 25-73 min, 20-38% B; 73-78 min, 38-46% B; 78-80 min, 46-49% B; 80-90 min, 49-56% B;90 to92min,56% -62% of B; 92-102 min, 62-75% B; 102-105 min, 75-100% B; the analysis results are shown in fig. 6, and it can be seen from the graph that a plurality of new ultraviolet absorption peaks appear in the saponin conversion group compared with the saponin control group and the strain control group, and the conversion product 9 is determined by comparing with 10 rare ginsenoside control products: ginsenoside 20- (S) -Rh 2 The method comprises the steps of carrying out a first treatment on the surface of the 10: ginsenoside 20- (R) -Rh 2 。
The preparation and analysis flow chart of the entire example 3 is shown in fig. 7.
Structural conversion reaction analysis of ginsenoside: the literature of the conversion of ginsenoside by a large number of natural microorganisms shows that the conversion of dammarane type tetracyclic triterpene saponins only occurs between protopanaxadiol type saponins or between protopanaxatriol type saponins, and no research on the fact that natural microorganisms can promote the mutual conversion between protopanaxadiol and triol type saponins is reported at present, based on the fact that the conversion substrate in the present invention is Rb only 1 The difference of the protopanaxadiol type saponin is known by comparing the structure: under the action of the bacterial enzyme system, ginsenoside Rb 1 Hydrolyzing the terminal glucose group at the C-20 position to obtain ginsenoside Rd, and hydrolyzing the terminal glucose group at the RdC-20 position to obtain ginsenoside F 2 Ginsenoside F 2 Isomerizing the chiral carbon (C-20) to form a pair of stereoisomers 20 (S) -Rh 2 And 20 (R) -Rh 2 The transformation path is as follows: the specific transformation reaction is shown in the following formula, and the enzyme system of trametes versicolor (trametes versicolor) has 3 activities of hydrolyzing glucose groups on C-20 positions of protopanaxadiol and triol type saponins, generating 20 (R/S) -ginsenoside stereoisomers, oxidizing C-20 hydroxyl groups to generate (20, 21) type and (20, 22) double bonds in principle.
The foregoing is only a specific embodiment of the invention to enable those skilled in the art to understand or practice the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown and described herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (10)
1. The trametes versicolor (trametes versicolor) is characterized in that the trametes versicolor is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.40999.
2. The method for converting trametes versicolor into ginsenoside Rb according to claim 1 1 Preparation of ginsenoside 20- (S) -Rh 2 、20-(R)-Rh 2 Is characterized in that: ginsenoside Rb by trametes versicolor 1 Fermenting, extracting to obtain total crude extract, and separating and purifying to obtain ginsenoside 20- (S) -Rh 2 、20-(R)-Rh 2 。
3. The method for converting trametes versicolor into ginsenoside Rb according to claim 2 1 Preparation of ginsenoside 20- (S) -Rh 2 、20-(R)-Rh 2 Is characterized by comprising the following steps:
(1) Culture of the strain: inoculating Coriolus versicolor trametes in culture medium, shake culturing T 1 d；
(2) Ginsenoside Rb 1 Is converted into: after the step (1) of shaking culture, adding substrate ginsenoside Rb 1 Then shake culture T is carried out 2 d, obtaining strain fermentation liquor;
(3) Extracting fermentation liquid: extracting the strain fermentation liquor by adopting an organic solvent, and concentrating the extracting solution under reduced pressure to obtain a total crude extract;
(4) And (3) separating and purifying: the total crude extract is separated and purified to obtain ginsenoside 20- (S) -Rh 2 、20-(R)-Rh 2 。
4. The method for converting trametes versicolor into ginsenoside Rb according to claim 3 1 Preparation of ginsenoside 20- (S) -Rh 2 、20-(R)-Rh 2 Wherein the medium in step (1) is PDB medium; the shake cultivation speed in the step (1) is 130-150 rmp/min, the cultivation temperature is 20-30 ℃, and T is not less than 3% 1 ≤5d。
5. The method for converting trametes versicolor into ginsenoside Rb according to claim 3 1 Preparation of ginsenoside 20- (S) -Rh 2 、20-(R)-Rh 2 Characterized in that the ginsenoside Rb in the step (2) 1 The preparation steps of (a) are as follows:
pulverizing Notoginseng radix main root, extracting with 50% -70% ethanol under reflux to obtain extractive solution, and purifying to obtain ginsenoside Rb 1 。
6. The method for converting trametes versicolor into ginsenoside Rb according to claim 3 1 Preparation of ginsenoside 20- (S) -Rh 2 、20-(R)-Rh 2 The method is characterized in that the shake cultivation speed in the step (2) is 130-150 rmp/min, the cultivation temperature is 20-30 ℃, and T is not less than 15 2 ≤20d。
7. The method for converting trametes versicolor into ginsenoside Rb according to claim 3 1 Preparation of ginsenoside 20- (S) -Rh 2 、20-(R)-Rh 2 The method is characterized in that the ginsenoside Rb added in the step (2) 1 The concentration is 0.01-0.1mg/mL.
8. The method for converting trametes versicolor into ginsenoside Rb according to claim 3 1 Preparation of ginsenoside 20- (S) -Rh 2 、20-(R)-Rh 2 Is characterized in that the organic solvent extraction step in the step (3) is as follows: adding water saturated n-butanol with 1-2 times of the liquid volume of the bacteria, centrifuging, retaining n-butanol layer extract, extracting for 3-5 times, mixing the extracts, and concentrating the extract under reduced pressure to obtain total crude extract.
9. The fermentation broth of claim 1 obtained by fermentation of trametes versicolor.
10. The method for converting ginsenoside Rb by trametes versicolor according to claim 1 1 Preparation of ginsenoside 20- (S) -Rh 2 And ginsenoside 20- (R) -Rh 2 Is used in the field of applications.
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