Method for preparing ginsenoside Rd-rich extract by fermenting pseudo-ginseng with paecilomyces farinosus

Abstract

The invention discloses a method for preparing an extract rich in ginsenoside Rd by fermenting pseudo-ginseng with paecilomyces farinosus, and belongs to the technical field of microbial fermentation. The method specifically comprises the following steps: crushing and sieving pseudo-ginseng main roots, pseudo-ginseng incisions and pseudo-ginseng stems and leaves, collecting powder with different granularities, and mixing the powder with different granularities to obtain pseudo-ginseng main root coarse powder, pseudo-ginseng incisions coarse powder and pseudo-ginseng stem and leaf coarse powder; preparing pseudo-ginseng powder; preparing a pseudo-ginseng culture medium; the preparation method comprises the following steps: co-culturing paecilomyces farinosus and a pseudo-ginseng culture medium to obtain a fermented suspension culture; the preparation method comprises the following steps: dissolving panax notoginseng into a suspension culture, evaporating the suspension culture to remove water to obtain panax notoginseng plastid, performing reflux extraction on the panax notoginseng plastid, concentrating an extracting solution to obtain extract, dissolving the extract with water, adsorbing an aqueous solution of the extract into resin, eluting, concentrating an eluent, and drying to obtain the extract rich in ginsenoside Rd. The method disclosed by the invention has the advantages of simplicity in operation, no environmental pollution and high efficiency.

Description

Technical Field

[0001] This invention relates to the field of microbial fermentation technology, specifically a method for preparing an extract rich in ginsenoside Rd from Panax notoginseng by fermenting Paecilomyces cerevisiae. Background Technology

[0002] powdery Penicillium ( Paecilomyces farinosus is Paecilomyces This genus of entomopathogenic fungi is mainly isolated from the bodies of lepidopteran pests such as pine caterpillars and pekinensis. Currently, it is primarily used for biological control; other applications have been reported, for example, the metabolites of *Paecilomyces oryzae* possess antibacterial, antioxidant, antitumor, and hypoglycemic activities, which can be used in the development of related biopharmaceutical products; in addition, *Paecilomyces oryzae* also has good application value in biotransformation, for example, it can effectively convert dehydroepiandrosterone (DHEA); using *Paecilomyces oryzae* to convert flavonoids can induce substrate glycosylation to obtain flavonoid glycosides.

[0003] Panax notoginseng (Burk.) FHChen is one of the oldest medicinal herbs in China, belonging to the genus Panax in the family Araliaceae. It is widely used to treat external bleeding, chest and abdominal pain, and swelling and pain from falls and injuries. Saponins are the representative substances and main active ingredients in Panax notoginseng. Products made from total Panax notoginseng saponins, such as Sanqi Tongshu capsules, Xuesaitong injection, and Xueshuantong capsules, have played an important role in the treatment of cardiovascular and cerebrovascular diseases. Panax notoginseng contains predominantly notoginseng saponins R1, ginsenoside Rg1, and ginsenoside Rb1, as well as rare saponins such as ginsenosides Rd, F2, Rk3, Rh2, Rh4, Rg5, and CK. These rare saponins also exhibit pharmacological activity in the treatment of diabetes, tumors, depression, hyperlipidemia, and obesity, making them a class of natural products with potential medicinal value. Panax notoginseng is a typical example of a Chinese herb used differently depending on whether it is raw or processed. Processed Panax notoginseng has the effect of nourishing blood and replenishing qi, which is closely related to the fact that the high-temperature environment of steaming promotes the conversion of saponins and increases the content of rare saponins. For example, after steaming, the content of ginsenosides Rh1, Rk3, Rh4, 20(S)-Rg3 and 20(R)-Rg3 in Panax notoginseng increases. However, the traditional steaming and boiling method has low efficiency in converting saponins, and the products are complex, the process is cumbersome, and the cost is high, making it difficult to use industrially.

[0004] Therefore, it is essential to develop an efficient fermentation method for Panax notoginseng saponins. Summary of the Invention

[0005] To address the problems of low conversion efficiency, complex products, cumbersome processes, and high costs associated with traditional saponin methods, this invention provides a method for preparing ginsenoside Rd-rich extracts from Panax notoginseng using Paecilomyces cerevisiae fermentation, specifically including the following steps: (1) The main root, cut end and stem and leaves of Panax notoginseng are crushed and sieved to collect powder of different particle sizes. The powder of different particle sizes are mixed to obtain coarse powder of main root, coarse powder of cut end and coarse powder of stem and leaves of Panax notoginseng.

[0006] (2) Panax notoginseng powder can be prepared by the following two methods: Method 1: Mix the coarse powder of Panax notoginseng main root with the coarse powder of Panax notoginseng stem and leaves to obtain Panax notoginseng powder.

[0007] Method 2: Mix the coarse powder from the cut end of Panax notoginseng with the coarse powder from the stems and leaves of Panax notoginseng to obtain Panax notoginseng powder.

[0008] (3) Add konjac powder, peptone, glucose, sodium chloride and purified water to the Panax notoginseng powder, sterilize, and obtain Panax notoginseng culture medium.

[0009] (4) Inoculate Paecilomyces powderyli into PDA plate medium, culture at constant temperature, pick colonies into PDA liquid medium and culture on a shaker, and then inoculate into the Panax notoginseng medium prepared in step (1) for dark culture to obtain the fermented suspension culture.

[0010] (5) Evaporate the water from the suspension culture to obtain Panax notoginseng mycelium. Reflux the Panax notoginseng mycelium with an ethanol aqueous solution with a volume percentage concentration of 70-80% three times. Concentrate the extract to obtain an extract. Dissolve the extract in water to obtain an extract aqueous solution. Adsorb the extract aqueous solution into D-101 macroporous resin, elute, concentrate the eluent, and dry to obtain an extract rich in ginsenoside Rd.

[0011] Preferably, the conditions for crushing and sieving in step (1) of the present invention are: the crushed raw material is passed through a 10-mesh sieve, a 20-mesh sieve and a 30-mesh sieve in sequence.

[0012] Preferably, in step (1) of the present invention, the mass ratio of 10-20 mesh powder and 20-30 mesh powder in the coarse powder of Panax notoginseng main root, coarse powder of Panax notoginseng cut, and coarse powder of Panax notoginseng stem and leaves is (0.5~1):(1~2); powder that can pass through a 10 mesh sieve but not a 20 mesh sieve is 10-20 mesh powder, and powder that can pass through a 20 mesh sieve but not a 30 mesh sieve is 20-30 mesh powder.

[0013] Preferably, in the Panax notoginseng powder prepared by method one in step (2) of the present invention, the mass ratio of Panax notoginseng root coarse powder to Panax notoginseng stem and leaf coarse powder is 10~7:1; in the Panax notoginseng powder prepared by method two, the mass ratio of Panax notoginseng cut coarse powder to Panax notoginseng stem and leaf coarse powder is 10~7:1.

[0014] Preferably, in step (3) of the present invention, the mass ratio of Panax notoginseng powder, konjac powder, peptone, glucose, sodium chloride and purified water in the Panax notoginseng culture medium is 100:(2~6):(2~6):(1~3):(0.5~1):(300~800).

[0015] Preferably, the isothermal culture in step (4) of the present invention is specifically: isothermal culture at room temperature until the diameter of the powdery Paecilomyces colony reaches 2.0~2.5cm; the shaking culture is specifically: shaking culture at 150r / min for 5~10 days; the dark culture is specifically: dark culture at 150~180r / min at room temperature for 25~35 days.

[0016] Preferably, the temperature for evaporating water in step (5) of the present invention is 60~70℃; in the reflux extraction, the mass ratio of ethanol aqueous solution to mycelium is 8~10:1, 5~8:1, and 5~8:1 respectively, and the extraction time is 2 hours each time.

[0017] Preferably, the conditions for concentrating the extract and dissolving it in water in step (5) of the present invention are: concentrating the extract at 55~65°C to a relative density of 1.1~1.3, and the mass ratio of extract to water is 1:8~10; the mass ratio of extract to dry resin is 1:8.

[0018] Preferably, the elution in step (5) of the present invention specifically involves: eluting with water until colorless; then eluting with an ethanol aqueous solution with a volume percentage concentration of 30%~40%, wherein the volume ratio of the ethanol aqueous solution with a volume percentage concentration of 30%~40% to the D-101 macroporous resin is 20~30:1; and then eluting with an ethanol aqueous solution with a volume percentage concentration of 95%, wherein the volume ratio of the ethanol aqueous solution with a volume percentage concentration of 95% to the D-101 macroporous resin is 20~30:1.

[0019] Preferably, in step (5) of the present invention, the concentrated eluent is concentrated at 55~65°C to a relative density of 1.1~1.3; the drying conditions are: drying at 65°C for 8 hours.

[0020] The powdery Paecilomyces described in this invention is a commercially available strain, purchased from the China Microbial Culture Collection website, number: bio-84377.

[0021] This invention provides a method for preparing ginsenoside Rd-rich extracts from Panax notoginseng using Paecilomyces cerevisiae fermentation, which has the following beneficial effects: (1) This invention utilizes the directional fermentation of Panax notoginseng matrix by Paecilomyces pulveratum to effectively transform common saponin components in Panax notoginseng and significantly increase the content of ginsenoside Rd and rare saponins such as F2, Rh4, 20(R)-Rg3, and CK.

[0022] (2) This invention can utilize multiple parts of Panax notoginseng, such as the main root, cut ends, stems and leaves, for fermentation. It can utilize traditional medicinal parts and also expand the use of non-main product resources such as stems and leaves, thereby improving the utilization rate of the whole Panax notoginseng plant, reducing raw material costs, and conforming to the development direction of efficient resource utilization.

[0023] (3) This invention uses fermentation and extraction purification processes, combined with the segmented elution strategy of D-101 macroporous resin, to effectively enrich the target saponin components, reduce impurity interference, and finally obtain an extract with high content of target components and clear components, which is suitable for the development and application of a variety of high value-added products such as pharmaceuticals, health products, and cosmetics.

[0024] (4) The process of this invention adopts conventional microbial fermentation and mature extraction technology, which is simple to operate, uses universal equipment, and is easy to scale up. Microbial transformation replaces high-temperature steaming, which is mild, energy-efficient, and allows for the recycling of ethanol. The overall process is clean and environmentally friendly, in line with the concept of green and sustainable development. Detailed Implementation

[0025] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0026] Example 1 A method for preparing ginsenoside Rd-rich extract from Panax notoginseng using Paecilomyces cerevisiae fermentation specifically includes the following steps: (1) The main root of three-year-old Panax notoginseng and its corresponding stems and leaves are crushed and sieved. The crushed raw materials are passed through a 10-mesh sieve, a 20-mesh sieve and a 30-mesh sieve in sequence. Powders of different particle sizes are collected and mixed to obtain coarse powder of Panax notoginseng main root and coarse powder of Panax notoginseng stems and leaves. The mass ratio of 10-20 mesh powder and 20-30 mesh powder in the coarse powder of Panax notoginseng main root and coarse powder of Panax notoginseng stems and leaves is 1:2.

[0027] (2) Preparation of Panax notoginseng powder: Panax notoginseng root coarse powder and Panax notoginseng stem and leaf coarse powder are mixed to obtain Panax notoginseng powder. The mass ratio of Panax notoginseng root coarse powder to Panax notoginseng stem and leaf coarse powder in the Panax notoginseng powder is 10:1.

[0028] (3) Place the Panax notoginseng powder in a culture bottle and add konjac powder, peptone, glucose, sodium chloride and purified water. The mass ratio of Panax notoginseng powder, konjac powder, peptone, glucose, sodium chloride and purified water is 100:4:4:2:1:500. Seal with a breathable plastic cap and sterilize at 121℃ for 30 min to obtain Panax notoginseng semi-solid suspension culture medium.

[0029] (4) Inoculate Paecilomyces powderylis into PDA agar plates and incubate at room temperature until the diameter of Paecilomyces powderylis colonies reaches 2.0 cm. Pick 1.0 cm of the colony. 2 The colonies were added to PDA liquid medium and cultured on a shaker at 150 r / min for 7 days. Then, they were inoculated into the Panax notoginseng medium prepared in step (1) and cultured at room temperature at 180 r / min in the dark for 30 days to obtain the fermented suspension culture.

[0030] (5) The suspension culture was dried at 65℃ to obtain Panax notoginseng mycelium. The mycelium was extracted three times by reflux with an ethanol aqueous solution with a volume percentage concentration of 70-80% (the mass ratio of ethanol aqueous solution to mycelium was 10:1, 8:1, and 8:1 respectively, and the extraction time was 2 hours each time). The extract was concentrated at 60℃ to a relative density of 1.15 to obtain an extract. The extract was dissolved in water (the mass ratio of extract to water was 1:10) to obtain an extract aqueous solution. The extract aqueous solution was adsorbed into D-101 macroporous resin (the mass ratio of extract to dry resin was 1:10). The eluent was eluted with water until colorless (ratio 1:8), then eluted with a 30% (v / v) ethanol aqueous solution (volume ratio of 30% ethanol aqueous solution to D-101 macroporous resin 20:1), then eluted with a 95% (v / v) ethanol aqueous solution (volume ratio of 95% ethanol aqueous solution to D-101 macroporous resin 20:1), the eluent was concentrated to a relative density of 1.15 (60℃), and dried at 65℃ for 8 hours to obtain ginsenoside extract.

[0031] Example 2 A method for preparing ginsenoside Rd-rich extract from Panax notoginseng using Paecilomyces cerevisiae fermentation specifically includes the following steps: (1) The main root of three-year-old Panax notoginseng and its corresponding stems and leaves are crushed and sieved. The crushed raw materials are passed through a 10-mesh sieve, a 20-mesh sieve and a 30-mesh sieve in sequence. Powders of different particle sizes are collected and mixed to obtain coarse powder of Panax notoginseng main root and coarse powder of Panax notoginseng stems and leaves. The mass ratio of 10-20 mesh powder and 20-30 mesh powder in the coarse powder of Panax notoginseng main root and coarse powder of Panax notoginseng stems and leaves is 0.5:2.

[0032] (2) Preparation of Panax notoginseng powder: Panax notoginseng root coarse powder and Panax notoginseng stem and leaf coarse powder are mixed to obtain Panax notoginseng powder. The mass ratio of Panax notoginseng root coarse powder to Panax notoginseng stem and leaf coarse powder in the Panax notoginseng powder is 7:1.

[0033] (3) Place the Panax notoginseng powder in a culture bottle and add konjac powder, peptone, glucose, sodium chloride and purified water. The mass ratio of Panax notoginseng powder, konjac powder, peptone, glucose, sodium chloride and purified water is 100:2:2:1:0.5:300. Seal with a breathable plastic cap and sterilize at 121℃ for 20 min to obtain Panax notoginseng semi-solid suspension culture medium.

[0034] (4) Inoculate Paecilomyces powderylis into PDA agar plates and incubate at room temperature until the diameter of Paecilomyces powderylis colonies reaches 2.5 cm. Pick 1.0 cm of the colony. 2 The colonies were added to PDA liquid medium and cultured on a shaker at 150 r / min for 5 days. Then, they were inoculated into the Panax notoginseng medium prepared in step (1) and cultured at room temperature at 150 r / min in the dark for 35 days to obtain the fermented suspension culture.

[0035] (5) The suspension culture was evaporated to dryness at 60℃ to obtain Panax notoginseng mycelium. The mycelium was then extracted three times by reflux with an 80% (v / v) ethanol aqueous solution (ethanol-to-mycelium mass ratio was 8:1, 5:1, and 5:1 respectively, with each extraction lasting 2 hours). The extract was concentrated to a relative density of 1.1 (55℃) to obtain an extract. The extract was dissolved in water (extract to water mass ratio was 1:8) to obtain an extract aqueous solution. This extract aqueous solution was adsorbed onto D-101 macroporous resin (extract to dry resin mass ratio was...). The mixture was eluted with water until colorless (1:8 ratio); then eluted with a 40% (v / v) ethanol aqueous solution, wherein the volume ratio of the 40% (v / v) ethanol aqueous solution to the D-101 macroporous resin was 30:1; then eluted with a 95% (v / v) ethanol aqueous solution, wherein the volume ratio of the 95% (v / v) ethanol aqueous solution to the D-101 macroporous resin was 30:1; the eluent was concentrated to a relative density of 1.1 (55℃), and dried at 65℃ for 8 hours to obtain the ginsenoside extract.

[0036] The content of each saponin in the ginsenoside extract obtained in this embodiment is similar to that in Example 1.

[0037] Example 3 A method for preparing ginsenoside Rd-rich extract from Panax notoginseng using Paecilomyces cerevisiae fermentation specifically includes the following steps: (1) The cut ends of three-year-old Panax notoginseng and their corresponding stems and leaves are crushed and sieved. The crushed raw materials are passed through a 10-mesh sieve, a 20-mesh sieve and a 30-mesh sieve in sequence to collect powders of different particle sizes. The powders of different particle sizes are mixed to obtain coarse powder of Panax notoginseng cut ends and coarse powder of Panax notoginseng stems and leaves. The mass ratio of 10-20 mesh powder and 20-30 mesh powder in the coarse powder of Panax notoginseng cut ends and coarse powder of Panax notoginseng stems and leaves is 1:1.

[0038] (2) Preparation of Panax notoginseng powder: Panax notoginseng cut coarse powder and Panax notoginseng stem and leaf coarse powder are mixed to obtain Panax notoginseng powder. The mass ratio of Panax notoginseng cut coarse powder to Panax notoginseng stem and leaf coarse powder in Panax notoginseng powder is 9:1.

[0039] (3) Place the Panax notoginseng powder in a culture bottle and add konjac powder, peptone, glucose, sodium chloride and purified water. The mass ratio of Panax notoginseng powder, konjac powder, peptone, glucose, sodium chloride and purified water is 100:6:6:3:0.75:800. Seal with a breathable plastic cap and sterilize at 121℃ for 40 min to obtain Panax notoginseng semi-solid suspension culture medium.

[0040] (4) Inoculate Paecilomyces powderylis into PDA agar plates and incubate at room temperature until the diameter of Paecilomyces powderylis colonies reaches 2.2 cm. Pick 1.0 cm of the colony. 2 The colonies were added to PDA liquid medium and cultured on a shaker at 150 r / min for 10 days. Then, they were inoculated into the Panax notoginseng medium prepared in step (1) and cultured at room temperature at 170 r / min in the dark for 25 days to obtain the fermented suspension culture.

[0041] (5) The suspension culture was dried at 70℃ to obtain Panax notoginseng mycelium. The mycelium was then extracted three times by reflux with a 75% (v / v) ethanol aqueous solution (ethanol-to-mycelium mass ratio was 10:1, 6:1, and 6:1 respectively, with each extraction lasting 2 hours). The extract was concentrated to a relative density of 1.3 (65℃) to obtain an extract. The extract was dissolved in water (extract to water mass ratio was 1:9) to obtain an extract aqueous solution. This extract aqueous solution was adsorbed onto D-101 macroporous resin (extract to dry resin mass ratio was...). The mixture was eluted with water until colorless (1:8 ratio); then eluted with a 35% (v / v) ethanol aqueous solution, wherein the volume ratio of the 35% (v / v) ethanol aqueous solution to the D-101 macroporous resin was 25:1; then eluted with a 95% (v / v) ethanol aqueous solution, wherein the volume ratio of the 95% (v / v) ethanol aqueous solution to the D-101 macroporous resin was 25:1; the eluent was concentrated to a relative density of 1.3 (65℃), and dried at 65℃ for 8 hours to obtain the ginsenoside extract.

[0042] The Panax notoginseng culture medium from step (3) of Examples 1 and 3 and the fermented suspension culture from step (4) were dried at 65°C, ground into powder, and passed through an 80-mesh sieve for later use. Approximately 1.00 g of the fine powder was placed in a 15 ml centrifuge tube, and an appropriate amount of pure methanol was added. The mixture was sonicated for 20 min. The centrifuge tube was then centrifuged at 4000 r / min for 10 min. The supernatant was transferred to a 50 ml volumetric flask and repeated three times. The mixture was then diluted to the mark with pure methanol, shaken well, and filtered through a 0.22 µm filter membrane. The contents of the monomeric saponins, such as Panax notoginseng saponin R1, ginsenosides Rg1, Rb1, Rc, Rb2, Rd, F2, Rh4, 20(R)-Rg3, and CK, in the filtrate were detected by HPLC.

[0043] The extracts and ginsenoside extracts that were not filtered through D-101 macroporous resin in steps (5) of Examples 1 and 3 were dried, ground, and 100 mg were accurately weighed into a 10 mL volumetric flask. Approximately 5.0 mL of chromatographically pure methanol was added, and the mixture was sonicated for 15 min to dissolve completely. After standing for 5 min, the volume was adjusted to the mark with chromatographically pure methanol. The solution was then filtered through a 0.22 μm filter membrane. The contents of monomeric saponins such as notoginsenoside R1, ginsenosides Rg1, Rb1, Rc, Rh1, Rb2, Rd, F2, Rh4, 20(R)-Rg3, and CK in the filtrate were determined by HPLC. Each sample was measured three times, and the average value of the three samples was taken as the saponin content of that sample.

[0044] The saponin content in each sample in Example 1 is detailed in Table 1.

[0045] Table 1 Note: — indicates not detected As shown in Table 1 above, in Example 1, after fermentation with Paecilomyces pilosa, an extract rich in ginsenoside Rd was obtained. The content of ginsenoside Rd accounted for 71.22% of the detected saponin content, and the total content of Rd, F2, Rh4, 20(R)-Rg3 and CK accounted for 95.41% of the detected saponin content, which significantly increased the content of ginsenoside Rd and rare saponins such as F2, Rh4, 20(R)-Rg3 and CK.

[0046] The saponin content in each sample in Example 3 is detailed in Table 2.

[0047] Table 2 Note: — indicates not detected As shown in Table 2 above, in Example 1, after fermentation with Paecilomyces pilosa, an extract rich in ginsenoside Rd was obtained. The content of ginsenoside Rd accounted for 71.84% of the detected saponin content, and the total content of Rd, F2, Rh4, 20(R)-Rg3 and CK accounted for 96.04% of the detected saponin content, which significantly increased the content of ginsenoside Rd and rare saponins such as F2, Rh4, 20(R)-Rg3 and CK.

[0048] In summary, co-culturing *Paecilomyces pilosa* in a semi-solid culture medium prepared by mixing coarse powder of Panax notoginseng main root (or coarse powder of Panax notoginseng cut) and coarse powder of Panax notoginseng stems and leaves in a certain mass ratio can significantly increase the Rd content in Panax notoginseng main root (or Panax notoginseng). Through ethanol extraction, eluting D-101 macroporous resin with different concentrations of ethanol can yield extracts rich in ginsenoside Rd, in which the proportion of ginsenoside Rd can reach more than 70%, and the proportion of rare saponins such as ginsenoside Rd, F2, Rh4, 20(R)-Rg3 and CK can reach more than 95%, providing a new approach for the rapid acquisition of rare saponin components.

[0049] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A method for preparing an extract rich in ginsenoside Rd using *Paecilomyces cerevisiae* fermentation of *Panax notoginseng*, characterized in that, Specifically, the following steps are included: (1) The main root, cut end and stem and leaves of Panax notoginseng are crushed and sieved to collect powder of different particle sizes. The powder of different particle sizes are mixed to obtain coarse powder of main root, coarse powder of cut end and coarse powder of stem and leaves of Panax notoginseng. (2) Panax notoginseng powder can be prepared by the following two methods: Method 1: Mix the coarse powder of Panax notoginseng main root with the coarse powder of Panax notoginseng stem and leaves to obtain Panax notoginseng powder; Method 2: Mix the coarse powder from the cut ends of Panax notoginseng with the coarse powder from the stems and leaves of Panax notoginseng to obtain Panax notoginseng powder; (3) Add konjac powder, peptone, glucose, sodium chloride and purified water to the Panax notoginseng powder, sterilize, and obtain Panax notoginseng culture medium; (4) Inoculate Paecilomyces powderylis into PDA plate medium, culture at constant temperature, pick colonies into PDA liquid medium and culture on a shaker, and then inoculate into the Panax notoginseng medium prepared in step (1) for dark culture to obtain the fermented suspension culture. (5) Evaporate the water from the suspension culture to obtain Panax notoginseng mycelium. Reflux the Panax notoginseng mycelium three times with an ethanol aqueous solution of 70-80% by volume. Concentrate the extract to obtain an extract. Dissolve the extract in water to obtain an extract aqueous solution. Adsorb the extract aqueous solution into D-101 macroporous resin, elute, concentrate the eluent, and dry to obtain an extract rich in ginsenoside Rd.

2. The method for preparing ginsenoside Rd-rich extract from Panax notoginseng using Paecilomyces cerevisiae fermentation according to claim 1, characterized in that, The conditions for crushing and sieving in step (1) are: the crushed raw material is passed through a 10-mesh sieve, a 20-mesh sieve and a 30-mesh sieve in sequence.

3. The method for preparing ginsenoside Rd-rich extract from Panax notoginseng using Paecilomyces cerevisiae fermentation according to claim 1, characterized in that, In step (1), the mass ratio of 10-20 mesh powder and 20-30 mesh powder in the coarse powder of Panax notoginseng main root, coarse powder of Panax notoginseng cut, and coarse powder of Panax notoginseng stem and leaves is (0.5~1):(1~2).

4. The method for preparing ginsenoside Rd-rich extract from Panax notoginseng using Paecilomyces cerevisiae fermentation according to claim 1, characterized in that, In step (2), the mass ratio of coarse powder of Panax notoginseng main root to coarse powder of Panax notoginseng stem and leaves in the Panax notoginseng powder prepared by method one is 10~7:1; in the Panax notoginseng powder prepared by method two, the mass ratio of coarse powder of Panax notoginseng cut ends to coarse powder of Panax notoginseng stem and leaves is 10~7:

1.

5. The method for preparing ginsenoside Rd-rich extract from Panax notoginseng using Paecilomyces cerevisiae fermentation according to claim 1, characterized in that, In step (3), the mass ratio of Panax notoginseng powder, konjac powder, peptone, glucose, sodium chloride and purified water in the Panax notoginseng culture medium is 100:(2~6):(2~6):(1~3):(0.5~1):(300~800).

6. The method for preparing ginsenoside Rd-rich extract from Panax notoginseng using Paecilomyces cerevisiae fermentation according to claim 1, characterized in that, The constant temperature culture in step (4) specifically refers to: constant temperature culture at room temperature until the diameter of the powdery Paecilomyces colony reaches 2.0~2.5cm; the shaker culture specifically refers to: shaker culture at 150r / min for 5~10 days; the dark culture specifically refers to: dark culture at room temperature at 150~180r / min for 25~35 days.

7. The method for preparing ginsenoside Rd-rich extract from Panax notoginseng using Paecilomyces cerevisiae fermentation according to claim 1, characterized in that, The temperature for evaporating water in step (5) is 60~70℃; in the reflux extraction, the mass ratio of ethanol aqueous solution to mycelium is 8~10:1, 5~8:1, and 5~8:1 respectively, and the extraction time is 2 hours each time.

8. The method for preparing ginsenoside Rd-rich extract from Panax notoginseng using Paecilomyces cerevisiae fermentation according to claim 1, characterized in that, The conditions for concentrating the extract and dissolving it in water in step (5) are: concentrating the extract at 55~65℃ until the relative density is 1.1~1.3, and the mass ratio of extract to water is 1:8~10; the mass ratio of extract to dry resin is 1:

8.

9. The method for preparing ginsenoside Rd-rich extract from Panax notoginseng using Paecilomyces cerevisiae fermentation according to claim 1, characterized in that, The elution in step (5) specifically involves: eluting with water until colorless; then eluting with an ethanol aqueous solution with a volume percentage concentration of 30%~40%, wherein the volume ratio of the ethanol aqueous solution with a volume percentage concentration of 30%~40% to the D-101 macroporous resin is 20~30:1; and then eluting with an ethanol aqueous solution with a volume percentage concentration of 95%, wherein the volume ratio of the ethanol aqueous solution with a volume percentage concentration of 95% to the D-101 macroporous resin is 20~30:

1.

10. The method for preparing ginsenoside Rd-rich extract from Panax notoginseng using Paecilomyces cerevisiae fermentation according to claim 1, characterized in that, The concentrated eluent in step (5) is concentrated at 55~65℃ to a relative density of 1.1~1.3; the drying conditions are: drying at 65℃ for 8 hours.

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