CN112691124A - Application of endophytic fungus A814 extract in preparation of antibacterial drugs - Google Patents
Application of endophytic fungus A814 extract in preparation of antibacterial drugs Download PDFInfo
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- CN112691124A CN112691124A CN202011581189.XA CN202011581189A CN112691124A CN 112691124 A CN112691124 A CN 112691124A CN 202011581189 A CN202011581189 A CN 202011581189A CN 112691124 A CN112691124 A CN 112691124A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
- C12P1/02—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using fungi
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- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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Abstract
The invention discloses application of an endophytic fungus A814 extract in preparation of antibacterial drugs. The invention discovers that the endophytic fungus Phomopsis liquidambaris A814 extract has obvious inhibiting effect on staphylococcus aureus and bacillus subtilis, and has good application and development prospect. Therefore, the invention provides a candidate drug for researching and developing a new antibacterial drug and provides a scientific basis for developing and utilizing natural active substances derived from endophytic fungi of plants.
Description
Technical Field
The invention belongs to the technical field of biological medicines, and particularly relates to an application of an extract of Aquilaria malabaricus endophytic fungus Phomopsis liquidamb aris A814 in preparation of antibacterial drugs.
Background
At present, more and more drug-resistant strains of pathogenic microorganisms are discovered in clinical treatment, the infection is more and more serious, and a plurality of traditional antibiotics basically lose the treatment efficacy, so that the search for novel antibacterial drugs is a key field of new drug development in the world. The endophytic fungi can produce secondary metabolites with abundant structures, the structures of the secondary metabolites are the same as or similar to certain metabolites of a host, and the secondary metabolites have potential medicinal values, and the compounds with new structures discovered from the secondary metabolites become important resources for discovering natural active substances. The active products discovered in endophytic fungi of plants in recent years mainly have the structural types of terpenes, alkaloids, quinones, flavonoids, isocoumarins, lactones, lignans, phenols, polypeptides, steroids and the like, the biological activities of the metabolites have the biological activities of resisting tumors, bacteria, oxidation, malaria, viruses, insects, enzyme inhibitors, regulating plant growth and the like, and the metabolites have good application prospects in various fields of medicines, agriculture, industry and the like.
Disclosure of Invention
The invention aims to provide application of an endophytic fungus Phomopsis liquidambaris A814 extract in preparing antibacterial drugs.
The invention discovers that the diameters of inhibition zones of staphylococcus aureus (staphylococcus aureus) and Bacillus subtilis (Bacillus subtilis) are respectively 12mm and 11mm when the content of an endophytic fungus Bacillus liquidambaris A814 extract in each filter paper sheet is 10 mu g, and the diameters of the inhibition zones of positive control medicament ampicillin on the 2 test strains are respectively 18mm and 20mm under the same concentration; the experimental results show that: the extract of the endophytic fungus Phomopsis liquidambaris A814 has significant antibacterial activity. Therefore, the extract of endophytic fungus Phomopsis liquidambaris A814 can be used for preparing antibacterial drugs.
The antibacterial drug is preferably a drug for resisting staphylococcus aureus or bacillus subtilis.
The second object of the present invention is to provide an antibacterial agent comprising an extract of endophytic fungus Phomopsis liquidambaris A814 as an active ingredient.
The antibacterial drug is preferably a drug for resisting staphylococcus aureus and bacillus subtilis.
Preferably, the endophytic fungus Phomopsis liquidambaris A814 extract is obtained by taking the endophytic fungus Phomopsis liquidambaris A814 as a fermentation strain, performing liquid fermentation to obtain a fermentation culture, separating a mycelium and a fermentation broth, extracting the fermentation broth with ethyl acetate, and concentrating the extraction liquid to obtain the endophytic fungus Phomopsis liquidambaris A814 extract.
Preferably, the liquid fermentation is to inoculate endophytic fungus Phomopsis Liquidambaris A814 into a fermentation culture medium, and carry out liquid fermentation culture at 28 ℃ and 120r/min to obtain a fermentation culture, wherein the fermentation culture medium is prepared by the following method per liter: decocting potato 200g in distilled water for 20min, filtering to obtain juice, and adding glucose 20g and KH2PO43g、MgSO40.75g, vitamin B110mg, adding water to 1L, and naturally adjusting pH to obtain fermentation medium.
The invention discovers that the endophytic fungus Phomopsis liquidambaris A814 extract has obvious inhibiting effect on staphylococcus aureus and bacillus subtilis, and shows that the endophytic fungus Phomopsis liquidambaris A814 extract has better antibacterial activity, can be used for preparing antibacterial drugs, and has good application and development prospects. Therefore, the invention provides a candidate drug for researching and developing a new antibacterial drug and provides a scientific basis for developing and utilizing natural active substances derived from endophytic fungi of plants.
Phomopsis Liquidambaris A814 of the present invention was deposited at the Guangdong province culture Collection (GDMCC) at 12/9/2020 with addresses: the Guangzhou city first furious Zhonglu No. 100 large yard No. 59 building No. 5 building, the preservation number is: GDMCC No: 3.691, which were available to the depository at the filing date of this application.
Detailed Description
The present invention will be better understood by those skilled in the art from the following examples. The examples are described only to illustrate the invention and should not be construed as limiting the invention as detailed in the claims.
Activation, fermentation culture and extract preparation of strain A841(Phomopsis liquidambia)
(1) Inoculating endophytic fungus Phomopsis liquidambaris A814 to a slant culture medium, and culturing at 28 ℃ for 5d to obtain an activated strain. The slant culture medium is obtained by the following method: decocting potato 200g in distilled water for 20min, filtering to obtain juice, and adding glucose 20g and KH2PO43g、MgSO40.75g, vitamin B110mg of agar and 20g of water, the solution is made up to 1L with natural pH and sterilized for later use.
(2) Inoculating the activated and cultured strain A814 in the step (1) into a seed culture medium, and culturing for 7d at 28 ℃ under the condition of 120r/min oscillation to obtain a seed solution, wherein the seed culture medium is obtained by the following method: decocting potato 200g in distilled water for 20min, filtering to obtain juice, and adding glucose 20g and KH2PO43g、MgSO40.75g, vitamin B110mg, adding water to 1L, naturally adjusting pH, and sterilizing.
(3) Inoculating the seed liquid obtained in the step (2) into a fermentation culture medium in an inoculation amount of 10% (v/v), and culturing at 28 ℃ and 120r/min for 7d to obtain a fermentation culture. The fermentation medium is the same as the seed culture medium.
(4) Preparing an extract: and (3) filtering the fermentation culture obtained in the step (3) to remove mycelia, extracting the filtrate for 3 times by using ethyl acetate, and concentrating the extract under reduced pressure to obtain the endophytic fungus Phomopsis liquidambaris A814 extract.
Example 1: determination of Activity of endophytic fungus Phomopsis liquidambaris A814 extract against Staphylococcus aureus and Bacillus subtilis
The extract of the endophytic fungus Phomopsis liquidambaris A814 was dissolved in DMSO and diluted to a concentration of 2mg/mL to give a sample dilution.
Blending Staphylococcus aureus or Bacillus subtilis in logarithmic growth phase with normal saline to give a solution with a concentration of 106Absorbing 1mL of the prepared bacterial liquid from the CFU/mL bacterial liquid into a culture dish, pouring LB culture medium cooled to a proper temperature, uniformly mixing, clamping a filter paper sheet with the thickness of 1.5mm and the diameter of 6mm by using a sterile forceps, placing the filter paper sheet on a bacterial-containing flat plate, and respectively dropwise adding 5 mu L of the sample diluent on the filter paper sheetAs a sample group, DMSO was used as a blank control group instead of the sample diluent, ampicillin was used as a positive control group, the plate was incubated at 37 ℃ for 24 hours, and the diameter of the zone of inhibition was measured by a cross measurement method. Repeating the steps for three times, and calculating the average value of the diameters of the inhibition zones.
The results of the experiment are shown in table 1:
table 1: inhibition of bacteria by extract of endophytic fungus Phomopsis liquidambaris A814 ((
n＝3)
The experimental results show that: the endophytic fungus Phomopsis liquidambaris A814 extract has obvious inhibition effect on staphylococcus aureus and bacillus subtilis and can be used for preparing medicines for resisting staphylococcus aureus and bacillus subtilis, so that the endophytic fungus Phomopsis liquidambaris A814 extract has wide application and development prospects in preparing antibacterial medicines.
The above is only a preferred embodiment of the present invention, and it should be noted that the above preferred embodiment should not be considered as limiting the present invention, and the protection scope of the present invention should be subject to the scope defined by the claims. It will be apparent to those skilled in the art that various modifications and adaptations can be made without departing from the spirit and scope of the invention, and these modifications and adaptations should be considered within the scope of the invention.
Claims (8)
1. Application of endophytic fungus Phomopsis liquidambaris A814 extract in preparing antibacterial drugs.
2. The application of claim 1, wherein the extract of endophytic fungus Phomopsis liquidambaris A814 is obtained by taking the endophytic fungus Phomopsis liquidambaris A814 as a fermentation strain, performing liquid fermentation to obtain a fermentation culture, separating mycelium and fermentation liquor, extracting the fermentation liquor with ethyl acetate, and concentrating the extract to obtain the extract of endophytic fungus Phomopsis liquidambaris A814.
3. The use according to claim 2, wherein the liquid fermentation is a fermentation culture obtained by inoculating the endophytic fungus Phomopsis liquidambaris A814 to a fermentation medium and performing liquid fermentation culture at 28 ℃ and 120r/min, wherein the fermentation medium is prepared by the following method per liter: decocting potato 200g in distilled water for 20min, filtering to obtain juice, and adding glucose 20g and KH2PO43 g、MgSO40.75g, vitamin B110mg, adding water to 1L, and naturally adjusting pH to obtain fermentation medium.
4. The use of claim 1, wherein said antibacterial agent is an agent against staphylococcus aureus or bacillus subtilis.
5. An antibacterial agent characterized by containing an extract of endophytic fungus Phomopsis liquidambaris A814 as an active ingredient.
6. The antibacterial drug according to claim 5, wherein the endophytic fungus Phomopsis liquidambaris A814 extract is obtained by taking the endophytic fungus Phomopsis liquidambaris A814 as a fermentation strain, performing liquid fermentation to obtain a fermentation culture, separating a mycelium and a fermentation broth, extracting the fermentation broth with ethyl acetate, and concentrating the extract to obtain the endophytic fungus Phomopsis liquidambaris A814 extract.
7. The antibacterial agent of claim 6, wherein the liquid fermentation is a fermentation culture obtained by inoculating Phomopsis liquidambaris A814 to a fermentation medium, and performing liquid fermentation at 28 ℃ and 120r/min, wherein each liter of the fermentation medium is prepared by the following method: decocting potato 200g in distilled water for 20min, and filtering to obtain juiceAdding 20g of glucose and KH2PO43 g、MgSO40.75g, vitamin B110mg, adding water to 1L, and naturally adjusting pH to obtain fermentation medium.
8. The antibacterial agent according to claim 5, wherein the antibacterial agent is an antibacterial agent against Staphylococcus aureus or Bacillus subtilis.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011581189.XA CN112691124A (en) | 2020-12-28 | 2020-12-28 | Application of endophytic fungus A814 extract in preparation of antibacterial drugs |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202011581189.XA CN112691124A (en) | 2020-12-28 | 2020-12-28 | Application of endophytic fungus A814 extract in preparation of antibacterial drugs |
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| CN112691124A true CN112691124A (en) | 2021-04-23 |
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| CN202011581189.XA Pending CN112691124A (en) | 2020-12-28 | 2020-12-28 | Application of endophytic fungus A814 extract in preparation of antibacterial drugs |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113956983A (en) * | 2021-11-15 | 2022-01-21 | 浙江中医药大学 | Atractylodes macrocephala endophytic fungus, fermentation product and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102154116A (en) * | 2010-12-22 | 2011-08-17 | 浙江大化生物科技股份有限公司 | Endophytic fungus Phomopsis sp. and use thereof |
| CN110862371A (en) * | 2019-11-19 | 2020-03-06 | 广东省微生物研究所（广东省微生物分析检测中心） | Polycyclic polyketone compound, preparation method thereof and application thereof in preparation of antibacterial drugs |
-
2020
- 2020-12-28 CN CN202011581189.XA patent/CN112691124A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102154116A (en) * | 2010-12-22 | 2011-08-17 | 浙江大化生物科技股份有限公司 | Endophytic fungus Phomopsis sp. and use thereof |
| CN110862371A (en) * | 2019-11-19 | 2020-03-06 | 广东省微生物研究所（广东省微生物分析检测中心） | Polycyclic polyketone compound, preparation method thereof and application thereof in preparation of antibacterial drugs |
Non-Patent Citations (1)
| Title |
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| 冉雪琴等: "罗汉松内生真菌的抑菌活性研究", 《中国畜牧兽医》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113956983A (en) * | 2021-11-15 | 2022-01-21 | 浙江中医药大学 | Atractylodes macrocephala endophytic fungus, fermentation product and application thereof |
| CN113956983B (en) * | 2021-11-15 | 2023-07-11 | 浙江中医药大学 | Endophytic fungus of bighead atractylodes rhizome, fermentation product and application thereof |
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