CN114621092A - Phenolic compound in mangrove plant-derived fungi and preparation method thereof - Google Patents
Phenolic compound in mangrove plant-derived fungi and preparation method thereof Download PDFInfo
- Publication number
- CN114621092A CN114621092A CN202210217654.4A CN202210217654A CN114621092A CN 114621092 A CN114621092 A CN 114621092A CN 202210217654 A CN202210217654 A CN 202210217654A CN 114621092 A CN114621092 A CN 114621092A
- Authority
- CN
- China
- Prior art keywords
- gradient
- component
- ethyl acetate
- compound
- reduced pressure
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C69/00—Esters of carboxylic acids; Esters of carbonic or haloformic acids
- C07C69/66—Esters of carboxylic acids having esterified carboxylic groups bound to acyclic carbon atoms and having any of the groups OH, O—metal, —CHO, keto, ether, acyloxy, groups, groups, or in the acid moiety
- C07C69/73—Esters of carboxylic acids having esterified carboxylic groups bound to acyclic carbon atoms and having any of the groups OH, O—metal, —CHO, keto, ether, acyloxy, groups, groups, or in the acid moiety of unsaturated acids
- C07C69/738—Esters of keto-carboxylic acids or aldehydo-carboxylic acids
-
- 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
- A61P31/10—Antimycotics
-
- C—CHEMISTRY; METALLURGY
- 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
-
- C—CHEMISTRY; METALLURGY
- 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
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/62—Carboxylic acid esters
Abstract
The invention relates to a phenolic compound in mangrove plant-derived fungi and a preparation method thereof, wherein the phenolic compound has a structure shown as a compound 1:
Description
Technical Field
The invention belongs to the field of secondary metabolites of mangrove fungi, and particularly relates to a phenolic compound in mangrove plant-derived fungi and a preparation method thereof.
Background
Mangrove plants grow in the tropical and subtropical intertidal zones, the living environment has the characteristics of high pressure, high salt, low oxygen and the like, so that the endophytic fungi have unique metabolic pathways, and further have the capacity of generating compounds with novel structures and various biological activities, and the metabolic products of the mangrove plants have various medicinal values of antibiosis, antitumor, immunoregulation, enzyme inhibition and the like and are potential microbial drug development resources, so the endophytic fungi of the mangrove plants become one of important resources for developing new drugs.
The method for artificially culturing and fermenting is adopted to obtain the secondary metabolite with important antibacterial activity from mangrove endophytic fungi, has the characteristics of environmental friendliness, sustainable development and the like, and can effectively solve key problems of medicine sources and the like in the process of medicine development, thereby having unique advantages. ML002 from Dothiorella sp
Disclosure of Invention
The fungus Dothiorella sp.ML002 is separated from mangrove plant melia azedarach, wherein the melia azedarach is a natural protection area of mangrove forest collected from Hongkong village of south east hai of south China sea in 2015 8 months. The fungus Dothiorella sp.ML002 is identified as the strain by morphology and molecular biology as the genus Microchaete; the sequence of the ITS region of this fungus has been submitted to NCBI (GenBank access No. MK131322). The "fungus Dothiorella sp.ml002" of the present invention has been disclosed in the inventor's previous research paper "Bioorganic Chemistry, 85, (2019), 382 + 385, Bioactive cell derivative isolated from the cultured-derivative fungal Dothiorella sp.ml002" and has been kept in the applicant's major laboratory of the chemical education department of tropical medicinal resources, and the public can obtain the "fungus Dothiorella sp.ml002" of the present invention according to the method described in the inventor's above research paper or purchase it from the applicant's major laboratory of the chemical education department of tropical medicinal resources, and the applicant can provide the "fungus Dothiorella sp.ml002" of the present invention to the public in the major laboratory of the chemical education department of tropical medicinal resources 20 years from the application date of the present invention.
The invention provides a phenolic compound or a pharmaceutically acceptable salt thereof, which is characterized in that the phenolic compound has a structure shown as a compound 1:
the invention provides a method for simultaneously preparing phenolic compounds 1 and 2, which is characterized by comprising the following steps:
(1) inoculating the fungus Dothiorella sp.ML002 into a fermentation culture medium, and performing static culture at 26-28 ℃ for 21-28 days to obtain a fermented product;
(2) extracting the fermentation product obtained in the step (1) with ethyl acetate with the volume of 1-2 times for 2-4 times, combining ethyl acetate phases, and concentrating under reduced pressure to obtain an extract;
(3) performing reduced pressure silica gel column chromatography on the extract obtained in the step (2), performing gradient elution by using petroleum ether-ethyl acetate as an eluent, wherein the elution gradient is respectively 100:0, 90:10, 80:20, 70:30, 60:40, 50:50, 40:60, 30:70, 20:80, 10:90 and 0:100, collecting two column volumes for each gradient, and dividing the column volumes into 7 components according to the polarity, wherein the gradient is 100: 0-90: 10 to obtain a component 1, the gradient is 80: 20-70: 30 to obtain a component 2, the gradient is 60:40 to obtain a component 3, the gradient is 50:50 to obtain a component 4, the gradient is 40: 60-30: 70 to obtain a component 5, the gradient is 20: 80-10: 90 to obtain a component 6, the gradient is 0:100 to obtain a component 7, the component 4 is subjected to normal phase silica gel column chromatography, the eluent is petroleum ether: eluting 4-5 column volumes with mixed solvent of ethyl acetate 8:1-3:1, concentrating under reduced pressure, and performing Sephadex LH-20 gel column chromatography, wherein the eluent is petroleum ether: CHCl3Eluting 4-5 column volumes with MeOH 2:1:1 mixed solvent, concentrating under reduced pressure, and performing High Performance Liquid Chromatography (HPLC) with Waters C18, 9.4 × 250mm, 7 μm, flow rate of 2mL/min, and mobile phase of MeOH H2O80: 20 to give compound 1 and compound 2.
Wherein the ratio of the eluent or the mobile phase is volume ratio; the fermentation medium is solid rice culture medium (formula is that 70ml water and 2.2g natural sea salt are added in per 60g rice).
Another embodiment of the present invention provides a crude extract fermented by the fungus Dothiorella sp.ML002, characterized in that the process for the preparation of the crude extract comprises the following steps:
(1) inoculating the fungus Dothiorella sp.ML002 into a fermentation culture medium, and performing static culture at 26-28 ℃ for 21-28 days to obtain a fermented product;
(2) and (2) extracting the fermentation product obtained in the step (1) with ethyl acetate with the volume of 1-2 times for 2-4 times, combining ethyl acetate phases, and then concentrating under reduced pressure to obtain an extract, namely the crude extract.
The fermentation medium is solid rice culture medium (formula is that 70ml water and 2.2g natural sea salt are added in per 60g rice).
The invention provides an anti-candida albicans drug, which is characterized in that the phenolic compounds 1 and/or 2, or pharmaceutically acceptable salts thereof or crude extracts thereof are used as effective components.
The anti-candida albicans drug provided by the invention also comprises other anti-candida albicans drugs; a pharmaceutically acceptable carrier or excipient may also be included.
Another embodiment of the invention provides the use of said phenolic compounds 1 and/or 2, or pharmaceutically acceptable salts thereof or the above crude extracts for the preparation of an anti-Candida albicans (Candida albicans) medicament.
The term "pharmaceutically acceptable salts" as used herein refers to non-toxic inorganic or organic acid and/or base addition salts, as described in Salt selection for basic drugs, int.J. pharm. (1986),33, 201-.
Drawings
FIG. 1 is a drawing of Compound 11H NMR chart;
FIG. 2 is a drawing of Compound 113C NMR chart;
FIG. 3 is a 135-DEPT diagram of Compound 1;
FIG. 4 is a HMQC plot for Compound 1;
FIG. 5 is a COSY diagram of Compound 1;
FIG. 6 is a HMBC diagram for Compound 1;
FIG. 7 is a HR-ESI-MS graph of Compound 1.
Detailed Description
In order to facilitate a further understanding of the invention, the following examples are provided to illustrate it in more detail. However, these examples are only for better understanding of the present invention and are not intended to limit the scope or the principle of the present invention, and the embodiments of the present invention are not limited to the following.
Example 1
(1) Preparing a fermentation medium: the formula is that 60g of rice, 70ml of water and 2.2g of natural sea salt are added into each conical flask. And (3) quenching at 120 ℃ for 25-30 minutes.
Inoculating the fungus Dothiorella sp.ML002 into a fermentation culture medium, and standing and culturing for 28 days at 26-28 ℃ to obtain a fermented product;
(2) extracting the fermentation product obtained in the step (1) with ethyl acetate with the volume 2 times of that of the fermentation product for 3 times, combining ethyl acetate phases, and then concentrating under reduced pressure to obtain an extract;
(3) performing reduced pressure silica gel column chromatography on the extract obtained in the step (2), performing gradient elution by using petroleum ether-ethyl acetate as an eluent, wherein the elution gradient is respectively 100:0, 90:10, 80:20, 70:30, 60:40, 50:50, 40:60, 30:70, 20:80, 10:90 and 0:100, collecting two column volumes for each gradient, and dividing the column volumes into 7 components according to the polarity, wherein the gradient is 100: 0-90: 10 to obtain a component 1, the gradient is 80: 20-70: 30 to obtain a component 2, the gradient is 60:40 to obtain a component 3, the gradient is 50:50 to obtain a component 4, the gradient is 40: 60-30: 70 to obtain a component 5, the gradient is 20: 80-10: 90 to obtain a component 6, the gradient is 0:100 to obtain a component 7, the component 4 is subjected to normal phase silica gel column chromatography, the eluent is petroleum ether: eluting 4-5 column volumes with mixed solvent of ethyl acetate 8:1-3:1, concentrating under reduced pressure, and performing Sephadex LH-20 gel column chromatography, wherein the eluent is petroleum ether: CHCl3Eluting 4-5 column volumes with MeOH 2:1:1 mixed solvent, concentrating under reduced pressure, and performing High Performance Liquid Chromatography (HPLC) with Waters C18, 9.4 × 250mm, 7 μm, flow rate of 2mL/min, and mobile phase of MeOH H2O80: 20 to give compound 1(5.0mg) and compound 2(3.6 mg).
The NMR structure confirmation data of compound 1 is as follows:
TABLE 1 preparation of Compound 11H(400MHz)and 13C (100MHz) NMR data (CDCl)3)
Compound 1: light brown powder; UV (MeOH) λ max (log ε)221(3.12),230(3.11),265(2.90),296(3.40) nm; IR (KBr) vmax 3289,1735,1602,1462,1365,1230cm-1；1H and 13C NMR data(CDCl3)see Table 1；HR-ESI-MS m/z 321.1340[M-H]-(calcd for C17H21O6,321.1338).
Process for preparation of Compound 21H/13C NMR was consistent with known reports of dothiorelone E (phytochem. lett.,22, 219) (2017)).
Example 2 antimicrobial testing
The compounds 1 to 2 and crude extracts of the present invention were tested against 5 pathogenic bacteria according to the microplate assay method described in the literature (Pierce c.g.; Uppuluri p.; teisan a.r.; Wormley jr.f.l.; Mowat e.; Ramage g.; Lopez-ribot j.l.nat. protoc.2008, 3, 1494-: staphylococcus aureus (ATCC 25923, Staphylococcus aureus), S.albus (ATCC 8032, Staphylococcus albus), Bacillus cereus (ATCC 14579, Bacillus cereus), Escherichia coli (ATCC 35218, Escherichia coli), Vibrio parahaemolyticus (ATCC17749, Vibrio parahaemolyticus), 1 strain of pathogenic fungus Candida albicans (ATCC 10231, Candida albicans). At the initial screening, at a concentration of 100. mu.g/mL, the compounds 1-2 and the crude extract showed antibacterial activity against Candida albicans (ATCC 10231, Candida albicans), especially the compound 2 and the crude extract showed significant antibacterial activity, and further dilution testing finally determined that the MIC value of the compound 2 against ATCC 10231 was 6.25. mu.g/mL, the MIC value of the crude extract against ATCC 10231 was 25.0. mu.g/mL, and the MIC value of the positive drug ketoconazole was 3.12. mu.g/mL.
Claims (8)
1. A phenolic compound or a pharmaceutically acceptable salt thereof, characterized in that the phenolic compound has the structure shown in compound 1:
2. a process for the simultaneous preparation of phenolic compounds 1 and 2, characterized in that it comprises the following steps:
(1) inoculating the fungus Dothiorella sp.ML002 into a fermentation culture medium, and performing static culture at 26-28 ℃ for 21-28 days to obtain a fermented product;
(2) extracting the fermentation product obtained in the step (1) with ethyl acetate with the volume of 1-2 times for 2-4 times, combining ethyl acetate phases, and concentrating under reduced pressure to obtain an extract;
(3) performing reduced pressure silica gel column chromatography on the extract obtained in the step (2), performing gradient elution by using petroleum ether-ethyl acetate as an eluent, wherein the elution gradient is respectively 100:0, 90:10, 80:20, 70:30, 60:40, 50:50, 40:60, 30:70, 20:80, 10:90 and 0:100, collecting two column volumes for each gradient, and dividing the column volumes into 7 components according to the polarity, wherein the gradient is 100: 0-90: 10 to obtain a component 1, the gradient is 80: 20-70: 30 to obtain a component 2, the gradient is 60:40 to obtain a component 3, the gradient is 50:50 to obtain a component 4, the gradient is 40: 60-30: 70 to obtain a component 5, the gradient is 20: 80-10: 90 to obtain a component 6, the gradient is 0:100 to obtain a component 7, the component 4 is subjected to normal phase silica gel column chromatography, the eluent is petroleum ether: eluting 4-5 column volumes with mixed solvent of ethyl acetate (8: 1-3: 1), concentrating under reduced pressure, and performing Sephadex LH-20 gel column chromatography with eluent petroleum ether: CHCl3Eluting 4-5 column volumes with MeOH 2:1:1 mixed solvent, concentrating under reduced pressure, and passing throughHPLC preparation with a chromatographic column of Waters C18, 9.4X 250mm, 7 μm, flow rate of 2mL/min, mobile phase of MeOH: H2O80: 20 to give compound 1 and compound 2; the ratio of the eluent or the mobile phase is volume ratio.
3. The method of claim 2, wherein the fermentation medium is a solid rice medium.
4. A crude extract fermented by the fungus Dothiorella sp.ML002, which is characterized in that the crude extract is prepared by the method of the steps (1) and (2) in claim 2, and comprises the following steps:
(1) inoculating a fungus Dothiorella sp.ML002 strain into a fermentation culture medium, and performing static culture at 26-28 ℃ for 21-28 days to obtain a fermentation product;
(2) extracting the fermentation product obtained in the step (1) with ethyl acetate with the volume of 1-2 times for 2-4 times, combining ethyl acetate phases, and then carrying out reduced pressure concentration to obtain an extract, namely the crude extract; the fermentation medium is a solid rice culture medium.
5. An anti-Candida albicans drug characterized by comprising the phenolic compounds 1 and/or 2 as described in any one of claims 1-2, or pharmaceutically acceptable salts thereof, or crude extract as described in claim 4 as an active ingredient.
6. The pharmaceutical of claim 5, further comprising an anti-Candida albicans agent.
7. Pharmaceutical according to any one of claims 5 to 6, characterized in that it further comprises a pharmaceutically acceptable carrier or excipient.
8. Use of phenolic compounds 1 and/or 2 according to any of claims 1 to 2, or of pharmaceutically acceptable salts thereof, or of a crude extract according to claim 4, for the preparation of a medicament against Candida albicans (Candida albicans).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210217654.4A CN114621092A (en) | 2022-03-08 | 2022-03-08 | Phenolic compound in mangrove plant-derived fungi and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210217654.4A CN114621092A (en) | 2022-03-08 | 2022-03-08 | Phenolic compound in mangrove plant-derived fungi and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114621092A true CN114621092A (en) | 2022-06-14 |
Family
ID=81899870
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210217654.4A Pending CN114621092A (en) | 2022-03-08 | 2022-03-08 | Phenolic compound in mangrove plant-derived fungi and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114621092A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115490661A (en) * | 2022-08-09 | 2022-12-20 | 海南师范大学 | Antioxidant active compound in mangrove-derived fungi and preparation method thereof |
-
2022
- 2022-03-08 CN CN202210217654.4A patent/CN114621092A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115490661A (en) * | 2022-08-09 | 2022-12-20 | 海南师范大学 | Antioxidant active compound in mangrove-derived fungi and preparation method thereof |
| CN115490661B (en) * | 2022-08-09 | 2023-09-08 | 海南师范大学 | Antioxidant active compound in mangrove-derived fungi and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107827805B (en) | Indole diterpenoid compound derived from mangrove plant xylocarpus meliloti fungi as well as preparation method and application thereof | |
| Biswas et al. | Endophytes producing podophyllotoxin from Podophyllum sp. and other plants: A review on isolation, extraction and bottlenecks | |
| CN106432262A (en) | Streptomyces-derived halogenated polyketide compound, preparation method and application | |
| CN106432168B (en) | The anti-vibrios reactive compound and preparation method thereof in mangrove cusp sea lotus endogenetic fungus source | |
| CN110257255B (en) | Benzopyrone derivative derived from endophytic fungi of mangrove cuspidate and lotus as well as preparation method and application thereof | |
| CN114621092A (en) | Phenolic compound in mangrove plant-derived fungi and preparation method thereof | |
| CN111909858B (en) | Mangrove endophytic fungus TGM112 secondary metabolite and application thereof | |
| Sun et al. | Effect of two ergosterol biosynthesis inhibitors on lycopene production by Blakeslea trispora | |
| CN110283728B (en) | Tetralone derivative derived from rosette valve and lotus endophytic fungi as well as preparation method and application thereof | |
| CN110330544B (en) | 4,4, 1-bicyclic steroid compound and preparation method and application thereof | |
| CN114437011B (en) | Chromone compound and preparation method and application thereof | |
| CN106432169B (en) | A kind of active isocoumarin class compound of anti-vibrios and its crystal | |
| CN115806881A (en) | Penicillium fungus and application thereof in preparation of antibacterial drugs | |
| CN106167494B (en) | Halogenation II type polyketides compound, preparation method and applications | |
| CN105274152B (en) | Curcumin biotransformation method, product and application | |
| CN111808015B (en) | Phenylalanine-derived cytochalasin as well as preparation method and application thereof | |
| CN108441427B (en) | Arthriospora fungi and pyridone alkaloid compound produced by same | |
| CN103145740A (en) | Sulfoxide alkaloid compound as well as preparation method and application for same | |
| CN107973803B (en) | Seven-membered lactonofuran derivative and preparation method and application thereof | |
| CN107686492A (en) | A kind of method of rhodioside in extraction purification zymotic fluid using macroporous absorbent resin | |
| CN107226800A (en) | A kind of xanthone classes compound and its preparation method of monocrystalline and the application as anti-Mycobacterium marinum medicine | |
| CN110172410A (en) | A kind of aphthopyrans ketone compound in marine fungi source, preparation method and applications | |
| CN107226801A (en) | A kind of xanthone classes compound and its preparation method of monocrystalline and the application as alpha-glucosidase restrainer | |
| CN111440200A (en) | Mixed source terpene alkaloid and anti-Zika virus application thereof | |
| CN114920792B (en) | Steroid compound and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |