Preparation of salvia miltiorrhiza endophytic bacterial lipopeptide and application of salvia miltiorrhiza endophytic bacterial lipopeptide in increasing content of pharmacodynamic substances in salvia miltiorrhiza

Abstract

The invention discloses preparation of salvia miltiorrhiza endophytic bacterial lipopeptide and application of the salvia miltiorrhiza endophytic bacterial lipopeptide in increasing the content of pharmacodynamic substances of salvia miltiorrhiza, and relates to the technical field of bioactive peptides. According to the invention, bacillus SNMB1 is used as a production strain of lipopeptide, and after fermentation, purification and separation, the salvia miltiorrhiza endophytic bacterial lipopeptide LP-1 is prepared. The radix salviae miltiorrhizae endophytic bacterial lipopeptide LP-1 prepared by the invention can effectively promote the quality improvement of radix salviae miltiorrhizae, especially the content of effective components of radix salviae miltiorrhizae. Specifically, after the salvia miltiorrhiza endophytic bacterial lipopeptide LP-1 treatment, the content of the salvianolic acid B in the salvia miltiorrhiza hairy roots can reach 131.25 mg / g, and compared with a blank treatment group, the content of the salvianolic acid B is increased by 134.84%.

Description

Technical Field

[0001] The invention relates to the technical field of bioactive peptides, in particular to the preparation of endophytic bacterial lipopeptides of salvia miltiorrhiza and the application of the lipopeptides in improving the content of effective substances of salvia miltiorrhiza. Background Art

[0002] Salvia miltiorrhiza is a traditional and commonly used bulk Chinese medicine in my country. It has a long history of medicinal use and is effective in the treatment of cardiovascular and cerebrovascular diseases. The market demand for Salvia miltiorrhiza ranks among the top bulk Chinese medicinal materials in my country. Salvianolic acid is an important water-soluble active ingredient in Salvia miltiorrhiza and is the main pharmacological substance used in the treatment of cardiovascular and other diseases. Salvianolic acid B is one of the important indicators for evaluating the quality of Salvia miltiorrhiza in the 2020 edition of the Chinese Pharmacopoeia and has rich biological activity. Therefore, how to increase the content of the active ingredients in Salvia miltiorrhiza is an important issue faced in the cultivation of Salvia miltiorrhiza.

[0003] Lipopeptide, also known as lipoacyl peptide, is a peptide formed by connecting fatty acids and peptide bonds with ester or amide bonds. Lipopeptides can be derived from plants, animals and microorganisms, among which microorganisms are the main ones, and most of the microorganisms are lipopeptides of bacterial origin, such as Bacillus, Pseudomonas, Actinomycetes, etc. Because of their amphiphilic structure, lipopeptides can reduce the surface tension of liquids and have good adsorption properties at the oil-water interface, thus showing good surface activity. This characteristic makes them have important application prospects in the field of food manufacturing. Lipopeptides also have broad-spectrum antibacterial activity, can destroy the cell membrane structure of bacteria, increase the permeability of bacteria, thereby causing the leakage of substances in bacteria, and thus killing bacteria. In terms of using lipopeptides to improve plant quality, CN116584484A discloses the use of lipopeptides to improve plant quality. Specifically, a composition composed of lipopeptides and adjuvants is used to improve plant quality, wherein the plants include vegetable plants, grains and fruit plants, etc. The lipopeptides include Bacillus ( Bacacillus ), Pseudomonas ( Pseudomonas ) and Streptomyces ( Streptomyces ) and other surfactants produced by iturin, fenmucin, lichenin, etc. However, the active ingredients in different types of plants are different, and the principles and methods for promoting the accumulation of active ingredients are naturally different. There are no reports on using lipopeptides to increase the content of active ingredients in Salvia miltiorrhiza. Summary of the invention

[0004] In view of the above-mentioned prior art, the purpose of the present invention is to provide the preparation of endophytic bacterial lipopeptides of Salvia miltiorrhiza and their application in increasing the content of effective substances in Salvia miltiorrhiza. The present invention uses endophytic bacteria Bacillus SNMB1 of Salvia miltiorrhiza as the production strain of lipopeptides, and obtains endophytic bacterial lipopeptide LP-1 of Salvia miltiorrhiza after fermentation, purification and separation. The endophytic bacterial lipopeptide LP-1 of Salvia miltiorrhiza prepared by the present invention can effectively promote the quality of Salvia miltiorrhiza, especially in increasing the content of effective ingredients in Salvia miltiorrhiza. Specifically, after being treated with endophytic bacterial lipopeptide LP-1 of Salvia miltiorrhiza, the content of salvianolic acid B in hairy roots of Salvia miltiorrhiza can reach 131.25 mg / g, and the content of salvianolic acid B is increased by 134.84% compared with the blank treatment group.

[0005] To achieve the above object, the present invention adopts the following technical solution: The first aspect of the present invention provides a method for preparing endophytic bacterial lipopeptide from Salvia miltiorrhiza, comprising the following steps: (1) Inoculating Bacillus SNMB1 into a fermentation medium, and obtaining a fermentation liquid after fermentation and cultivation; (2) After the fermentation liquid is centrifuged for the first time, the supernatant is collected and the pH is adjusted to 1.5-2.5. After standing, a suspension is obtained. After the suspension is centrifuged for the second time, the precipitate is collected, the precipitate is mixed with methanol, the pH is adjusted to 7.0, stirred, and centrifuged for the third time, and the supernatant is collected, which is the crude lipopeptide extract. (3) The crude lipopeptide extract and methanol are mixed to obtain a crude lipopeptide extract, and the crude lipopeptide extract is purified by high performance liquid chromatography to obtain endophytic bacterial lipopeptides from Salvia miltiorrhiza.

[0006] Preferably, in step (1), the deposit number of the Bacillus SNMB1 is CGMCC No. 25566, which is recorded in patent CN 116716210A.

[0007] Preferably, in step (1), the fermentation medium comprises: 200.0 g potato, 20.0 g glucose, and 1.0 L water.

[0008] Preferably, in step (1), the fermentation temperature is 23-27° C., the fermentation speed is 160-200 r / min, and the fermentation time is 2-4 days.

[0009] Preferably, in step (2), during the first centrifugation and the second centrifugation, the centrifugal speed is 4500-5500 r / min, and the centrifugal time is 25-35 min.

[0010] Preferably, in step (2), the standing temperature is 23-27° C. and the standing time is 1.5-2.5 h.

[0011] Preferably, in step (2), the pH of the supernatant is adjusted to 1.5-2.5 using dilute hydrochloric acid, wherein the mass fraction of the dilute hydrochloric acid is 10%; and the pH is adjusted to 7.0 using sodium hydroxide solution, wherein the mass fraction of the sodium hydroxide solution is 5%.

[0012] Preferably, in step (2), the solid-liquid ratio of precipitate to methanol is (8-12) mg:1 mL.

[0013] Preferably, in step (2), the stirring temperature is 23-27° C., the stirring speed is 250-350 r / min, and the stirring time is 4-6 h.

[0014] Preferably, in step (2), in the third centrifugation, the centrifugal speed is 8000-12000 r / min, and the centrifugal time is 25-35 min.

[0015] Preferably, in step (3), the concentration of the crude lipopeptide extract in the crude lipopeptide extract is 90-110 mg / mL.

[0016] Preferably, in step (3), the specific operation of purification by high performance liquid chromatography is: using a semi-preparative high performance liquid chromatography column and adopting a mobile phase for gradient elution to separate the endophytic bacterial lipopeptides of Salvia miltiorrhiza, wherein the injection volume is 50-100 μL, the column temperature is 25°C, the detection wavelength is 210 nm, and the flow rate is 2.0 mL / min.

[0017] Further preferably, the mobile phase consists of mobile phase A and mobile phase B, mobile phase A is 0.1% acetic acid aqueous solution, and mobile phase B is 0.1% acetic acid acetonitrile.

[0018] Further preferably, the gradient elution process is: 0-1.0 min, 90% mobile phase A; 1.0-14 min, 90%-0% mobile phase A; 14-17 min, 0% mobile phase A; 17-18 min, 0%-90% mobile phase A; 18-20 min, 90% mobile phase A.

[0019] The second aspect of the present invention provides a lipopeptide of endophytic bacteria of Salvia miltiorrhiza prepared by the above-mentioned preparation method, wherein the chemical structural formula of the lipopeptide of endophytic bacteria of Salvia miltiorrhiza is as follows: .

[0020] The third aspect of the present invention provides the use of the above-mentioned endophytic bacterial lipopeptide of Salvia miltiorrhiza in the following 1) and / or 2): 1) Increase the content of effective substances in Salvia miltiorrhiza; 2) Increase the number of Salvia miltiorrhiza hairy roots.

[0021] Preferably, the medicinal ingredient of Salvia miltiorrhiza is salvianolic acid B.

[0022] Beneficial effects of the present invention: The present invention uses the endophytic bacteria Bacillus SNMB1 of Salvia miltiorrhiza as the production strain of lipopeptides, and obtains the endophytic bacteria lipopeptide LP-1 of Salvia miltiorrhiza after fermentation, purification and separation. The endophytic bacteria lipopeptide LP-1 of Salvia miltiorrhiza prepared by the present invention can effectively promote the quality of Salvia miltiorrhiza, especially in improving the content of the effective components of Salvia miltiorrhiza. Specifically, after being treated with the endophytic bacteria lipopeptide LP-1 of Salvia miltiorrhiza, the content of salvianolic acid B in the hairy roots of Salvia miltiorrhiza can reach 131.25 mg / g, and the content of salvianolic acid B is increased by 134.84% compared with the blank treatment group. In addition, the present invention can reduce the use of chemicals by using the endophytic bacteria lipopeptide of Salvia miltiorrhiza to increase the content of the effective components of Salvia miltiorrhiza and promote the growth of the hairy roots of Salvia miltiorrhiza, and has important application value in promoting the increase of production and quality of Salvia miltiorrhiza and the sustainable development of green ecology. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 : 3D chromatogram of the crude lipopeptide extract obtained in Example 1; Figure 2 : The purification chromatogram of the endophytic bacterial lipopeptide of Salvia miltiorrhiza obtained in Example 1; Figure 3 : Mass spectrum of the endophytic bacterial lipopeptide of Salvia miltiorrhiza obtained in Example 1; Figure 4 : Growth morphology of Salvia miltiorrhiza hairy roots in the test group and blank control group in the experimental example. DETAILED DESCRIPTION

[0024] It should be noted that the following detailed descriptions are illustrative and are intended to provide further explanation of the present application. Unless otherwise specified, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art to which the present application belongs.

[0025] In order to enable those skilled in the art to more clearly understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below in conjunction with specific embodiments.

[0026] The experimental materials used in the embodiments of the present invention are all conventional experimental materials in the art and can be purchased through commercial channels.

[0027] Example 1: Preparation method of endophytic bacterial lipopeptide from Salvia miltiorrhiza (1) Inoculate Bacillus SNMB1 into a fermentation medium, and ferment at 25°C and 180 r / min for 3 days to obtain a fermentation liquid; The composition of the fermentation medium is 200.0 g potato, 20.0 g glucose, and 1.0 L water; (2) The fermentation broth was centrifuged at 5000 r / min for 30 min to remove the bacteria, and the supernatant after centrifugation was collected. The pH of the supernatant was adjusted to 2.0 with dilute hydrochloric acid, and the suspension was allowed to stand at 25°C for 2 h to obtain a suspension; the suspension was centrifuged at 5000 r / min for 30 min, and the precipitate after centrifugation was collected. The precipitate and methanol were mixed at a ratio of 10 mg:1 mL, and the pH was adjusted to 7.0 with sodium hydroxide solution. The mixture was stirred at 25°C and 300 r / min for 5 h, and then centrifuged at 10000 r / min for 30 min. The supernatant after centrifugation was collected, which was the crude lipopeptide extract; (3) The crude lipopeptide extract was dissolved in methanol to obtain a crude lipopeptide extract, wherein the concentration of the crude lipopeptide extract was 100 mg / mL, and the crude lipopeptide extract was purified by high performance liquid chromatography to obtain an endophytic bacterial lipopeptide from Salvia miltiorrhiza, which was named Lipopeptide 1 (LP1 for short); During the purification process, semi-preparative HPLC (Waters C18 column, 10 mm x 250 mm x 0.5 μm) was used to separate the endophytic bacterial lipopeptides of Salvia miltiorrhiza after gradient elution with the mobile phase. The injection volume was 50-100 μL, the column temperature was 25°C, the detection wavelength was 210 nm, and the flow rate was 2.0 mL / min. The mobile phase consisted of mobile phase A and mobile phase B. Mobile phase A was 0.1% acetic acid aqueous solution, and mobile phase B was 0.1% acetic acid acetonitrile. The gradient elution process was: 0-1.0 min, 90% mobile phase A; 1.0-14 min, 90%-0% mobile phase A; 14-17 min, 0% mobile phase A; 17-18 min, 0%-90% mobile phase A; 18-20 min, 90% mobile phase A.

[0028] The structural formula of the endophytic bacterial lipopeptide of Salvia miltiorrhiza prepared in this example is as follows: .

[0029] The crude lipopeptide extract obtained in this example was subjected to chromatographic analysis, and the results were as follows: Figure 1 As shown, the structure of the endophytic bacterial lipopeptide of Salvia miltiorrhiza prepared in this example was characterized, and the results are as follows Figure 2-Figure 3 shown.

[0030] Depend on Figure 1 It can be seen that the crude lipopeptide extract contains a series of lipopeptide metabolites. Figure 2 It can be seen that the obtained substance is pure and the retention time t R It is 6.02min.

[0031] Depend on Figure 3 It can be seen that the quasi-molecular ion peak of the endophytic bacterial lipopeptide of Salvia miltiorrhiza prepared in this example is m / z 1059.5726 [M+H+ ],1081.5542[M+Na + ], and then determine its molecular formula is C 60 H 78 N 10 O 15 .

[0032] Embodiment 2: (1) Inoculate Bacillus SNMB1 into a fermentation medium, and ferment at 23°C and 160 rpm for 2 days to obtain a fermentation broth; The composition of the fermentation medium is 200.0 g potato, 20.0 g glucose, and 1.0 L water; (2) The fermentation broth was centrifuged at 4500 r / min for 25 min to remove the bacteria, and the supernatant after centrifugation was collected. The pH of the supernatant was adjusted to 1.5 with dilute hydrochloric acid, and the mixture was allowed to stand at 25°C for 2 h to obtain a suspension; the suspension was centrifuged at 4500 r / min for 25 min, and the precipitate after centrifugation was collected. The precipitate and methanol were mixed at a ratio of 8 mg:1 mL, and the pH was adjusted to 7.0 with sodium hydroxide solution. The mixture was stirred at 23°C and 250 r / min for 4 h, and then centrifuged at 8000 r / min for 25 min. The supernatant after centrifugation was collected, which was the crude lipopeptide extract; (3) The crude lipopeptide extract was dissolved in methanol to obtain a crude lipopeptide extract, wherein the concentration of the crude lipopeptide extract was 90 mg / mL. The crude lipopeptide extract was purified by high performance liquid chromatography to obtain an endophytic bacterial lipopeptide from Salvia miltiorrhiza, which was named Lipopeptide 1 (LP1 for short). During the purification process, semi-preparative HPLC (Waters C18 column, 10 mm x 250 mm x 0.5 μm) was used to separate the endophytic bacterial lipopeptides of Salvia miltiorrhiza after gradient elution with the mobile phase. The injection volume was 50-100 μL, the column temperature was 25°C, the detection wavelength was 210 nm, and the flow rate was 2.0 mL / min. The mobile phase consisted of mobile phase A and mobile phase B. Mobile phase A was 0.1% acetic acid aqueous solution, and mobile phase B was 0.1% acetic acid acetonitrile. The gradient elution process was: 0-1.0 min, 90% mobile phase A; 1.0-14 min, 90%-0% mobile phase A; 14-17 min, 0% mobile phase A; 17-18 min, 0%-90% mobile phase A; 18-20 min, 90% mobile phase A.

[0033] Embodiment 3: (1) Inoculate Bacillus SNMB1 into a fermentation medium, and ferment at 27°C and 200 r / min for 4 days to obtain a fermentation liquid; The composition of the fermentation medium is 200.0 g potato, 20.0 g glucose, and 1.0 L water; (2) The fermentation broth was centrifuged at 5500 r / min for 35 min to remove the bacteria, and the supernatant after centrifugation was collected. The pH of the supernatant was adjusted to 2.5 with dilute hydrochloric acid, and the suspension was allowed to stand at 25°C for 2 h to obtain a suspension; the suspension was centrifuged at 5500 r / min for 35 min, and the precipitate after centrifugation was collected. The precipitate and methanol were mixed at a ratio of 12 mg:1 mL, and the pH was adjusted to 7.0 with sodium hydroxide solution. The mixture was stirred at 27°C and 350 r / min for 6 h, and then centrifuged at 12000 r / min for 35 min. The supernatant after centrifugation was collected, which was the crude lipopeptide extract; (3) The crude lipopeptide extract was dissolved in methanol to obtain a crude lipopeptide extract, wherein the concentration of the crude lipopeptide extract was 110 mg / mL. The crude lipopeptide extract was purified by high performance liquid chromatography to obtain an endophytic bacterial lipopeptide from Salvia miltiorrhiza, which was named Lipopeptide 1 (LP1 for short). During the purification process, semi-preparative HPLC (Waters C18 column, 10 mm x 250 mm x 0.5 μm) was used to separate the endophytic bacterial lipopeptides of Salvia miltiorrhiza after gradient elution with the mobile phase. The injection volume was 50-100 μL, the column temperature was 25°C, the detection wavelength was 210 nm, and the flow rate was 2.0 mL / min. The mobile phase consisted of mobile phase A and mobile phase B. Mobile phase A was 0.1% acetic acid aqueous solution, and mobile phase B was 0.1% acetic acid acetonitrile. The gradient elution process was: 0-1.0 min, 90% mobile phase A; 1.0-14 min, 90%-0% mobile phase A; 14-17 min, 0% mobile phase A; 17-18 min, 0%-90% mobile phase A; 18-20 min, 90% mobile phase A.

[0034] Test example: The endophytic bacterial lipopeptide of Salvia miltiorrhiza prepared in Example 1 was used in the Salvia miltiorrhiza hairy root culture system. After the culture, the content of salvianolic acid B therein was detected. The specific steps were as follows: (1) A Salvia miltiorrhiza hairy root system was placed in a 6.7v culture medium, and the ratio of the amount of the Salvia miltiorrhiza hairy root system to the 6.7v culture medium was 1 g fresh weight: 100 mL. After culturing at 10 r / min and 28°C for 30 days, the Salvia miltiorrhiza endophytic bacterial lipopeptide prepared in Example 1 was added to a final concentration of 20 μmol / L as a test group (LP1 group). A blank control group (CK group) was also set up. The blank control group was not added with the Salvia miltiorrhiza endophytic bacterial lipopeptide. After culturing for 15 days, the growth status of the Salvia miltiorrhiza hairy roots in the test group and the blank control group was as follows: Figure 4 As shown; (2) The hairy root materials of the test group and the blank control group were collected and freeze-dried at -80°C to obtain dry matter; 0.5 g of dry matter was accurately weighed, and 35 mL of 60% methanol solution was added and ultrasonic-assisted extraction was performed for 40 min to obtain an extract. The extract was filtered through a filter membrane, and the content of salvianolic acid B was determined by high performance liquid chromatography. The results are shown in Table 1; Among them, Ultra-High Performance Liquid Chromatography- Q ExactivePlus Orbitrap Mass Spectrometer (UPLC-QE-MS / MS) liquid chromatography-mass spectrometry was used for analysis.

[0035] Liquid phase conditions: Thermo Hypersil GOLD aQ C18 chromatographic column (2.1mm×100mm, 1.9μm) was used, column temperature: 35℃; mobile phases consisted of mobile phase A and mobile phase B, where mobile phase A was 0.1% acetic acid solution and mobile phase B was 0.1% acetic acid acetonitrile solution, gradient elution program: 0-0.5 min, 90% mobile phase A; 0.5-7 min, 90%-0% mobile phase A; 7-8.5 min, 0% mobile phase A; 8.5-8.6 min, 90% mobile phase A; 8.6-10min, 90% mobile phase A, injection volume: 1μL, flow rate: 0.3mL / min. Mass spectrometry conditions: positive ion mode: spray voltage: 3.8kv; sheath gas: 40; auxiliary gas: 10; ion transfer tube temperature: 350°C. Resolution: 17500; Microscan: 1; AGCtarget: 2e5; Normalized collision energy: 50. Negative ion mode: Spray voltage: 2.9kV; Sheath gas: 40; Auxiliary gas: 0; Ion transfer tube temperature: 350°C. Resolution: 17500; Microscan: 1; AGC target: 2e 5 ; Normalized collision energy: 50. Full scan mode (Full MS) was used, positive and negative ions were detected simultaneously, the scan range was set to 70-1050 m / z, and the resolution was 35000.

[0036] Table 1 Contents of salvianolic acid B in hairy roots of Salvia miltiorrhiza in different treatment groups Depend on Figure 4It can be seen that the number of hairy roots with the addition of endophytic bacterial lipopeptide from Salvia miltiorrhiza is much higher than that without the addition of endophytic bacterial lipopeptide from Salvia miltiorrhiza. It can be seen that the treatment with endophytic bacterial lipopeptide from Salvia miltiorrhiza can significantly increase the number of hairy roots of Salvia miltiorrhiza. As can be seen from Table 1, the content of salvianolic acid B in the hairy roots without the addition of endophytic bacterial lipopeptide from Salvia miltiorrhiza is 55.59 mg / g, while the content of salvianolic acid B in the hairy roots of Salvia miltiorrhiza after 15 days of treatment with endophytic bacterial lipopeptide from Salvia miltiorrhiza is 131.25 mg / g. Compared with the blank treatment group, the content of salvianolic acid B in the hairy roots of Salvia miltiorrhiza treated with endophytic bacterial lipopeptide from Salvia miltiorrhiza is increased by 2.35 times. It can be seen that endophytic bacterial lipopeptide from Salvia miltiorrhiza can significantly increase the content of effective substances of Salvia miltiorrhiza, especially, the content of salvianolic acid B in Salvia miltiorrhiza.

[0037] The above description is only the preferred embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A method for preparing endophytic bacterial lipopeptide from Salvia miltiorrhiza, characterized in that: The following steps are involved: (1) Inoculating Bacillus SNMB1 into a fermentation medium, and obtaining a fermentation liquid after fermentation and cultivation; (2) After the fermentation liquid is centrifuged for the first time, the supernatant is collected and the pH is adjusted to 1.5-2.

5. After standing, a suspension is obtained. After the suspension is centrifuged for the second time, the precipitate is collected, the precipitate is mixed with methanol, the pH is adjusted to 7.0, stirred, and centrifuged for the third time, and the supernatant is collected, which is the crude lipopeptide extract. (3) The crude lipopeptide extract and methanol are mixed to obtain a crude lipopeptide extract, and the crude lipopeptide extract is purified by high performance liquid chromatography to obtain endophytic bacterial lipopeptides from Salvia miltiorrhiza.

2. The method for preparing the endophytic bacterial lipopeptide of Salvia miltiorrhiza according to claim 1, characterized in that: In step (1), the deposit number of the Bacillus SNMB1 is CGMCC No.25566.

3. The method for preparing the endophytic bacterial lipopeptide of Salvia miltiorrhiza according to claim 1, characterized in that: In step (1), the composition of the fermentation medium is: 200.0 g potato, 20.0 g glucose, and 1.0 L water.

4. The method for preparing the endophytic bacterial lipopeptide of Salvia miltiorrhiza according to claim 1, characterized in that: In step (1), the fermentation temperature is 23-27°C, the fermentation speed is 160-200 r / min, and the fermentation time is 2-4 days.

5. The method for preparing the endophytic bacterial lipopeptide of Salvia miltiorrhiza according to claim 1, characterized in that: In step (2), during the first and second centrifugation processes, the centrifugal speed is 4500-5500 r / min, and the centrifugal time is 25-35 min; during the third centrifugation, the centrifugal speed is 8000-12000 r / min, and the centrifugal time is 25-35 min.

6. The method for preparing the endophytic bacterial lipopeptide of Salvia miltiorrhiza according to claim 1, characterized in that: In step (3), the concentration of the crude lipopeptide extract in the crude lipopeptide extract is 90-110 mg / mL.

7. The method for preparing the endophytic bacterial lipopeptide of Salvia miltiorrhiza according to claim 1, characterized in that: In step (3), the specific operation of purification by high-performance liquid chromatography is as follows: using a semi-preparative high-performance liquid chromatography column and adopting a mobile phase for gradient elution to separate the endophytic bacterial lipopeptides of Salvia miltiorrhiza, wherein the injection volume is 50-100 μL, the column temperature is 25°C, the detection wavelength is 210 nm, and the flow rate is 2.0 mL / min.

8. The method for preparing the endophytic bacterial lipopeptide of Salvia miltiorrhiza according to claim 7, characterized in that: In step (3), the mobile phase consists of mobile phase A and mobile phase B, mobile phase A is 0.1% acetic acid aqueous solution, and mobile phase B is 0.1% acetic acid acetonitrile; the gradient elution process is: 0-1.0 min, 90% mobile phase A; 1.0-14 min, 90%-0% mobile phase A; 14-17 min, 0% mobile phase A; 17-18 min, 0%-90% mobile phase A; 18-20 min, 90% mobile phase A.

9. The endophytic bacterial lipopeptide of Salvia miltiorrhiza obtained by the preparation method according to any one of claims 1 to 8, characterized in that: The structure of the endophytic bacterial lipopeptide of Salvia miltiorrhiza is as follows: 。 10. Use of the endophytic bacterial lipopeptide of Salvia miltiorrhiza according to claim 9 in the following 1) and / or 2): 1) Increase the content of effective substances in Salvia miltiorrhiza; 2) Increase the number of Salvia miltiorrhiza hairy roots.

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