Prussella adamsii Sys-Gs-003 strain for producing ginsenoside Rh2 and application of Prussella adamsii Sys-Gs-003 strain
The preparation of ginsenoside Rh2 by fermentation with *Priestella auriculi* strain Sys-Gs-003 solves the problems of high production cost and complex process in existing technologies, and realizes large-scale production with high efficiency and low cost.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-09
- Publication Date
- 2026-04-07
- Estimated Expiration
- Not applicable · inactive patent
AI Technical Summary
Existing technologies are insufficient for the efficient and low-cost production of ginsenoside Rh2, and are limited by the complexity of growth conditions and extraction processes, making it difficult to meet the needs of large-scale industrial production.
Ginsenoside Rh2 was prepared by fermentation using *Priscilla argentea* strain Sys-Gs-003. The optimized fermentation medium and conditions were: 5 g/L yeast extract, 8 g/L peptone, 3 g/L sodium chloride, pH 7.0, 37 ℃, 150 r/min, and fermentation for 7 days.
This method enables efficient and low-cost large-scale fermentation production of ginsenoside Rh2, avoiding the high cost and complexity of traditional methods and making it suitable for industrial applications.
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Figure CN121801745A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to a strain of Priestia aryabhattai. and use, the strain can ferment to prepare ginsenoside Rh2, belongs to the field of microbial technology. BACKGROUND
[0002] Ginsenoside is the main active medicinal component of ginseng. Due to the different types and contents of ginsenoside monomers contained, the pharmacological effects of various monomer saponins are different, so separating effective monomers and clarifying the pharmacological activities of each monomer have become a research hotspot in the field. At present, more than 60 known ginsenosides have been found, and each has a unique medicinal function, mainly divided into two categories: ① protopanaxadiol (PPD) including ginsenoside Ra1, Ra2, Rb1, Rb2, Rb3, Rc, Rd, Rg3 and Rh2, etc.; ② protopanaxatriol (PPT) including ginsenoside Re, Rg1, Rg2, Rf and Rh1, etc.
[0003] Most of the saponins in ginseng are Rb1, Rb2, Rc, Rd, Re and Rg1. Ginsenoside Rh2 is a very rare ginsenoside in ginseng. Ginsenoside Rh2 is one of the most important anti-tumor active substances in ginseng, which has the effects of inhibiting tumor cell growth, inhibiting telomerase activity of tumor cells, inducing tumor cell apoptosis, anti-tumor cell metastasis, reversing abnormal differentiation of tumor cells, etc., thereby having good anti-tumor and preventing and treating tumor recurrence and metastasis effects. In clinical practice, ginsenoside Rh2 combined with radiotherapy and chemotherapy and other treatment methods can enhance the effect of radiotherapy and chemotherapy in cancer treatment. In addition, ginsenoside Rh2 also has a series of physiological functions such as anti-allergy, improving immunity, anti-fatigue and anti-inflammatory.
[0004] At present, there are mainly two ways to produce ginsenoside Rh2. The first way is to use ginsenoside Rh2 as raw material, which is extracted from ginseng, and to hydrolyze ginsenoside Rh2 by acid hydrolysis, alkali hydrolysis, enzymatic hydrolysis and microbial fermentation. Due to the long growth cycle of ginseng, the harsh growth conditions, the low content of ginsenoside Rh2, the complex extraction process and the low yield of ginsenoside Rh2, the production cost of ginsenoside Rh2 is extremely high, which is difficult to meet the requirements of large-scale industrial production. (2) The precursor of ginsenoside Rh2, protopanaxadiol, is used as a substrate to produce ginsenoside Rh2 by chemical modification. The chemical synthesis route is complex, the cost is high, the conversion rate is low, the stereoselectivity is poor and a large amount of by-products are generated. In recent years, it has been rarely used. The above two production methods have their own disadvantages, so seeking other production methods is a research hotspot in the field. There is no related report on the direct microbial fermentation production of rare ginsenoside Rh2. SUMMARY
[0005] The present application solves the problems in the background art and provides a strain of Priestia aryabhattai Sys-Gs-003 capable of producing ginsenoside Rh2. The strain can be used for fermentation to prepare ginsenoside Rh2.
[0006] A new strain is screened in the present application, which is named Sys-Gs-003 and is a kind of Priestia aryabhattai. The strain is preserved in China Center for Type Culture Collection, Wuhan University, Wuhan, China, with a postal code of 430072, and the preservation date is December 4, 2025, and the preservation number is CCTCC NO: M20252779. The 16S rDNA sequence of the strain is shown as SEQ ID NO. 1.
[0007] The above-mentioned strain of Priestia aryabhattai Sys-Gs-003 can be used for fermentation to prepare ginsenoside Rh2. The microbial fermentation conditions are: 5 g / L of yeast powder, 8 g / L of proteose peptone, 3 g / L of sodium chloride, pH 7.0; the culture conditions are: 37 ℃, 150 r / min, and fermentation culture for 7 days.
[0008] Compared with the prior art, the present application has the following advantages: the strain of Priestia aryabhattai Sys-Gs-003 provided by the present application can produce ginsenoside Rh2. The strain provided by the present application can be used for large-scale fermentation culture in a short period of time, and the fermentation cost is low, and it is not limited by time, season and other conditions, which has obvious advantages. Meanwhile, the present application also provides the optimized microbial fermentation conditions and the formula of the culture medium, and the yield of ginsenoside Rh2 is high. BRIEF DESCRIPTION OF DRAWINGS
[0009] Figure 1 HPLC detection chart of ginsenoside Rh2 in standard sample;
[0010] Figure 2 HPLC detection chart of ginsenoside Rh2 in fermentation medium of the strain of Priestia alba Sys-Gs-003. DETAILED DESCRIPTION
[0011] The application will be described in detail below with specific examples, but the scope of protection of the application is not limited to the following examples.
[0012] Isolation and culture of strains
[0013] In this embodiment, the root tissue of fresh wild ginseng collected from Changbai Mountain was surface sterilized with 75% ethanol for 20 min, then washed with sterile water for 3-5 times, and put into a pre-sterilized mortar, 5 mL of sterile water was added to grind into a white suspension. 300 μL of the suspension was taken to each LB solid medium plate, and evenly coated with a sterile swab. After air-drying, the plate was sealed with a sealing film and incubated at 37°C in an incubator for 3-5 days. The growth of bacteria was observed and recorded in time, and the bacteria were repeatedly purified.
[0014] At the same time, the stem was washed, the necrotic surface hard part was removed, and then cut into 1 cm 2 thin pieces. The pieces were first soaked in 75% alcohol for 3-5 min, then washed with sterile water for 2-3 times to remove the residual alcohol, and then soaked in sterile water for about 5 min. After taking out, the surface moisture was absorbed with sterile filter paper. The treated pieces were placed on the LB solid medium plate, 5 pieces on each plate, sealed with a sealing film, and incubated at 37°C in an incubator for 3-5 days. The growth of bacteria was observed and recorded in time, and the bacteria were repeatedly purified.
[0015] The purified bacteria on the plate were picked into a 1.5 mL EP tube containing 600 μL of LB liquid medium, sealed with a sealing film, and then incubated at 37°C in a 190 r / min shaking incubator for 1 day. An equal volume of 50% glycerol was added, mixed well, and stored in a -80°C ultra-low temperature refrigerator. Each strain was stored for at least 3 copies.
[0016] 10 μL of the stored strain was inoculated into 5 mL of LB liquid medium, incubated at 37°C in a 150 r / min shaking incubator for 24 h, and 3 mL of the activated bacterial solution was inoculated into 100 mL of LB liquid medium for fermentation culture at 37°C in a 150 r / min shaking incubator for 7 d. After the above fermentation culture, the fermentation broth was collected.
[0017] Strain screening
[0018] The HPLC detection of ginsenoside Rh2 in the filtered fermentation broth sample of each strain is performed, and the detection result is compared with the standard data, so as to preliminarily determine whether the strain has the ability to produce ginsenoside Rh2.
[0019] After the above detection, a strain of Priestia aryabhattai producing ginsenoside Rh2 is screened, which is named as Sys-Gs-003 strain. The HPLC detection is shown in Figure 2 The HPLC detection conditions are as follows:
[0020] The HPLC detection conditions are as follows: the chromatograph is Agilent 1290 chromatographic system, the ODS-C18 reversed-phase column (4.6 mm x 250 mm, 5 μm, Agilent), the mobile phase is acetonitrile: water = 65:35; the standard concentration is 100 μg / mL, the flow rate is 1 mL / min, the ultraviolet wavelength is set to 203 nm, the column temperature is 20℃, and the detection result is shown in Figure 1 The comparison of Figure 1 and Figure 2 can show that the fermentation broth of the Sys-Gs-003 strain contains ginsenoside Rh2.
[0021] Sequencing and sequence alignment and analysis
[0022] The strain is sent to a biotechnology company for sequencing, and the 16S rDNA sequence of the strain is shown as SEQ ID NO. 1. The sequencing result is subjected to Blast similarity analysis in the NCBI nucleic acid database, and through Blast sequence alignment and phylogenetic tree analysis, the 16S rDNA sequence of the strain has a similarity of 99% with Priestia aryabhattai (GeneBank number: NR_115953.1), and it is determined to be a bacterium of Priestia aryabhattai.
[0023] Preservation of the strain:
[0024] The strain is preserved in the China Center for Type Culture Collection, address: China, Wuhan, Wuhan University; postcode 430072, preservation date is December 4, 2025, and preservation number is CCTCC NO: M20252779.
[0025] Optimization of fermentation conditions
[0026] The fermentation medium components of the microorganism must be able to meet the needs of rapid growth and reproduction of the strain, and the optimal fermentation culture conditions can ensure the accumulation of secondary metabolites. The present application preliminarily optimizes the fermentation medium components and culture conditions of the Sys-Gs-003 strain, and the optimized medium formula is: yeast powder 5 g / L, peptone 8 g / L, sodium chloride 3 g / L, pH 7.0; the culture conditions are: 37 DEG C, 150 r / min, and fermentation culture for 7 days.
Claims
1. A strain of *Priestia aryabhattai* Sys-Gs-003 that produces ginsenoside Rh2, with accession number CCTCC NO: M20252779, and its 16S rDNA sequence is shown in SEQ ID NO.
1.
2. The use of the *Priscilla argentea* strain Sys-Gs-003 according to claim 1, characterized in that: Ginsenoside Rh2 was prepared by fermentation.
3. The use according to claim 2, characterized in that: The fermentation method was as follows: fermentation temperature 37 ℃, rotation speed 150 r / min, fermentation culture for 7 days.
4. The use according to claim 2, characterized in that: The culture medium used in the fermentation process has the following formula: yeast powder 5 g / L, peptone 8 g / L, sodium chloride 3 g / L, pH 7.0.