CN118126007A - Microorganism secondary metabolite and preparation method and application thereof - Google Patents
Microorganism secondary metabolite and preparation method and application thereof Download PDFInfo
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- 229930000044 secondary metabolite Natural products 0.000 title claims abstract description 25
- 238000002360 preparation method Methods 0.000 title claims abstract description 15
- 244000005700 microbiome Species 0.000 title claims abstract description 11
- 150000001875 compounds Chemical class 0.000 claims abstract description 19
- 230000000813 microbial effect Effects 0.000 claims abstract description 13
- 239000000284 extract Substances 0.000 claims abstract description 10
- 241000228143 Penicillium Species 0.000 claims abstract description 9
- 241000228168 Penicillium sp. Species 0.000 claims abstract description 5
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 15
- 238000004440 column chromatography Methods 0.000 claims description 12
- 239000012071 phase Substances 0.000 claims description 12
- 238000000926 separation method Methods 0.000 claims description 11
- NOTFZGFABLVTIG-UHFFFAOYSA-N Cyclohexylethyl acetate Chemical compound CC(=O)OCCC1CCCCC1 NOTFZGFABLVTIG-UHFFFAOYSA-N 0.000 claims description 9
- 241000209094 Oryza Species 0.000 claims description 9
- 235000007164 Oryza sativa Nutrition 0.000 claims description 9
- 235000009566 rice Nutrition 0.000 claims description 9
- 239000001963 growth medium Substances 0.000 claims description 8
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 7
- 239000000287 crude extract Substances 0.000 claims description 6
- WGLUMOCWFMKWIL-UHFFFAOYSA-N dichloromethane;methanol Chemical compound OC.ClCCl WGLUMOCWFMKWIL-UHFFFAOYSA-N 0.000 claims description 6
- 239000000499 gel Substances 0.000 claims description 6
- 239000007791 liquid phase Substances 0.000 claims description 6
- 238000002791 soaking Methods 0.000 claims description 6
- 241000588747 Klebsiella pneumoniae Species 0.000 claims description 5
- 241000577483 Salmonella enterica subsp. enterica serovar Paratyphi B Species 0.000 claims description 5
- 238000000746 purification Methods 0.000 claims description 5
- 238000009210 therapy by ultrasound Methods 0.000 claims description 5
- 239000007787 solid Substances 0.000 claims description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 4
- NSGDYZCDUPSTQT-UHFFFAOYSA-N N-[5-bromo-1-[(4-fluorophenyl)methyl]-4-methyl-2-oxopyridin-3-yl]cycloheptanecarboxamide Chemical compound Cc1c(Br)cn(Cc2ccc(F)cc2)c(=O)c1NC(=O)C1CCCCCC1 NSGDYZCDUPSTQT-UHFFFAOYSA-N 0.000 claims description 3
- 238000004821 distillation Methods 0.000 claims description 3
- 238000010898 silica gel chromatography Methods 0.000 claims description 3
- XDTMQSROBMDMFD-UHFFFAOYSA-N Cyclohexane Chemical compound C1CCCCC1 XDTMQSROBMDMFD-UHFFFAOYSA-N 0.000 claims description 2
- 239000000022 bacteriostatic agent Substances 0.000 claims description 2
- 238000012258 culturing Methods 0.000 claims description 2
- 239000012153 distilled water Substances 0.000 claims description 2
- 239000003112 inhibitor Substances 0.000 claims description 2
- 238000011081 inoculation Methods 0.000 claims description 2
- 230000001954 sterilising effect Effects 0.000 claims description 2
- 238000004659 sterilization and disinfection Methods 0.000 claims description 2
- 230000000844 anti-bacterial effect Effects 0.000 abstract description 6
- 239000003814 drug Substances 0.000 abstract description 6
- 238000000855 fermentation Methods 0.000 abstract description 5
- 230000004151 fermentation Effects 0.000 abstract description 5
- 229940079593 drug Drugs 0.000 abstract description 4
- 238000002474 experimental method Methods 0.000 abstract description 3
- 239000002207 metabolite Substances 0.000 abstract description 2
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 9
- OKKJLVBELUTLKV-MZCSYVLQSA-N Deuterated methanol Chemical compound [2H]OC([2H])([2H])[2H] OKKJLVBELUTLKV-MZCSYVLQSA-N 0.000 description 8
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 7
- 241000233866 Fungi Species 0.000 description 5
- 238000005160 1H NMR spectroscopy Methods 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 3
- 241000222122 Candida albicans Species 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 3
- 229940095731 candida albicans Drugs 0.000 description 3
- 239000013641 positive control Substances 0.000 description 3
- 241000588724 Escherichia coli Species 0.000 description 2
- 241000282414 Homo sapiens Species 0.000 description 2
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 230000000975 bioactive effect Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000002114 high-resolution electrospray ionisation mass spectrometry Methods 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000012453 solvate Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 235000017166 Bambusa arundinacea Nutrition 0.000 description 1
- 235000017491 Bambusa tulda Nutrition 0.000 description 1
- 229930186147 Cephalosporin Natural products 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- GSDSWSVVBLHKDQ-JTQLQIEISA-N Levofloxacin Chemical compound C([C@@H](N1C2=C(C(C(C(O)=O)=C1)=O)C=C1F)C)OC2=C1N1CCN(C)CC1 GSDSWSVVBLHKDQ-JTQLQIEISA-N 0.000 description 1
- 244000082204 Phyllostachys viridis Species 0.000 description 1
- 235000015334 Phyllostachys viridis Nutrition 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- 108010059993 Vancomycin Proteins 0.000 description 1
- 125000000217 alkyl group Chemical group 0.000 description 1
- 230000008485 antagonism Effects 0.000 description 1
- 229940124350 antibacterial drug Drugs 0.000 description 1
- 239000011425 bamboo Substances 0.000 description 1
- 230000001851 biosynthetic effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000001460 carbon-13 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 229940124587 cephalosporin Drugs 0.000 description 1
- 150000001780 cephalosporins Chemical class 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000009509 drug development Methods 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- RFHAOTPXVQNOHP-UHFFFAOYSA-N fluconazole Chemical compound C1=NC=NN1CC(C=1C(=CC(F)=CC=1)F)(O)CN1C=NC=N1 RFHAOTPXVQNOHP-UHFFFAOYSA-N 0.000 description 1
- 229960004884 fluconazole Drugs 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000009036 growth inhibition Effects 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000001678 irradiating effect Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 229960003376 levofloxacin Drugs 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005360 mashing Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 239000010413 mother solution Substances 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 125000001424 substituent group Chemical group 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000004879 turbidimetry Methods 0.000 description 1
- 229960003165 vancomycin Drugs 0.000 description 1
- MYPYJXKWCTUITO-UHFFFAOYSA-N vancomycin Natural products O1C(C(=C2)Cl)=CC=C2C(O)C(C(NC(C2=CC(O)=CC(O)=C2C=2C(O)=CC=C3C=2)C(O)=O)=O)NC(=O)C3NC(=O)C2NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(CC(C)C)NC)C(O)C(C=C3Cl)=CC=C3OC3=CC2=CC1=C3OC1OC(CO)C(O)C(O)C1OC1CC(C)(N)C(O)C(C)O1 MYPYJXKWCTUITO-UHFFFAOYSA-N 0.000 description 1
- MYPYJXKWCTUITO-LYRMYLQWSA-O vancomycin(1+) Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=C2C=C3C=C1OC1=CC=C(C=C1Cl)[C@@H](O)[C@H](C(N[C@@H](CC(N)=O)C(=O)N[C@H]3C(=O)N[C@H]1C(=O)N[C@H](C(N[C@@H](C3=CC(O)=CC(O)=C3C=3C(O)=CC=C1C=3)C([O-])=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)O2)=O)NC(=O)[C@@H](CC(C)C)[NH2+]C)[C@H]1C[C@](C)([NH3+])[C@H](O)[C@H](C)O1 MYPYJXKWCTUITO-LYRMYLQWSA-O 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to the field of medicines, in particular to a preparation method of two novel microbial secondary metabolites and a medicine. The invention separates and extracts 2 new microorganism secondary metabolites from Penicillium fungus (Penicillium sp.) fermentation culture, namely compounds C1 and C2, and the structures of the compounds are shown in figure 1. C1 and C2 are 2 new metabolites discovered from Penicillium for the first time, the invention enriches the diversity of secondary metabolites of Penicillium, and antibacterial activity experiments show that compounds C1 and C2 have certain antibacterial effect.
Description
Technical Field
The invention belongs to the technical field of medicines, and relates to a microbial secondary metabolite, a preparation method and application thereof.
Background
Fungi are used as an independent biological group, are various in variety and function, play an important role in life and production processes, and are a natural medicine resource treasury. The secondary metabolites thereof provide a large amount of bioactive substances for human beings, and the extraction of bioactive components by using the metabolites of fungi is increasingly receiving attention from those skilled in the relevant arts.
Penicillium is one of the most common fungi, ranging from soil to vegetation, to air, to indoor environments and various foods. Penicillium has been developed worldwide for its biosynthetic potential, and many compounds isolated from various species of the Penicillium genus have shown in vitro as well as in vivo growth inhibition properties against different human disorders. Therefore, the penicillium has potential drug development and application prospect in terms of the genus. The research on the rhizosphere microorganisms of the medicinal plants can help to understand the medicinal material components of the medicinal plants and the biosynthesis mechanism thereof, improve the green cultivation of the Chinese medicinal materials, prevent and treat the quality degradation of the medicinal materials, alleviate the problem of deficient medicinal plant resources and the like, so that the method has important significance; penicillium sp. belongs to the common fungus, but no isolation of the compounds according to the invention from this fungus is known.
Disclosure of Invention
The invention aims to provide a microbial secondary metabolite, a preparation method and application thereof.
In order to achieve the above purpose, the invention adopts the technical scheme that:
A microbial secondary metabolite, which is a compound C1 and C2 shown in the following structural formula,
And pharmaceutically acceptable salts, solvates or isomers of compounds C1 and C2, e.g.,
Wherein: r 1、R2 is H, C 1-C4 carbonyl, hydroxy, substituted or unsubstituted C 1-C4 alkyl, and the substituent is methyl.
The preparation method of the microbial secondary metabolite comprises the following steps:
(1) Fermenting and producing strains:
Culturing Penicillium sp strain on rice culture medium at 26-28deg.C for 30-35 days to obtain fermented product; soaking the fermented product with ethyl acetate, performing ultrasonic treatment, and performing reduced pressure distillation to obtain crude extract, dissolving and purifying the crude extract to obtain total extract;
(2) Separation and purification of the compound:
the total extract is subjected to silica gel column chromatography by a cyclohexane-ethyl acetate system (100:1-1:100), crude fractions with cyclohexane-ethyl acetate (V/V) of 80-70:20-30 are collected, dichloromethane-methanol (1:1) is used as a mobile phase, gel column chromatography separation is carried out first, ODS column chromatography separation is carried out, and a dual-wavelength full-preparation liquid phase is used for separation and purification by using methanol-water (30%) as a mobile phase to obtain a compound C1;
Collecting cyclohexane-ethyl acetate (V/V) of 60-50:40-50, separating with dichloromethane-methanol (1:1) as mobile phase, separating with gel column chromatography, separating with ODS column chromatography, and separating and purifying with dual-wavelength full-prepared liquid phase with methanol-water (10%) as mobile phase to obtain compound C2.
And (3) dissolving the crude extract in the step (1) by using 90% methanol-water, repeatedly extracting by using cyclohexane with equal volume after dissolving, combining the extracting solutions, and concentrating under reduced pressure again to obtain the total extract.
The solid rice culture medium is prepared from 35-45g rice, 50-55mL distilled water, and is filled into 200-300mL conical flask, pH is natural, and sterilization is carried out at 121deg.C for 30-60min.
The inoculation concentration of the strain (Penicillium sp.) in step (1) is: 10 5-107/mL.
And (3) soaking the fermented product in the step (1) for 24-48 hours by using ethyl acetate, and performing ultrasonic treatment for 1-2 hours. The dual wavelength refers to: 210-254nm.
The application of the microbial secondary metabolite in preparing a bacteriostatic agent.
The application of the microbial secondary metabolite in preparing an inhibitor of Klebsiella pneumoniae and salmonella paratyphi B.
The invention has the advantages that:
The invention provides application of two novel microbial secondary metabolites C1 and C2 and pharmaceutically acceptable salts, solvates or isomers thereof or pharmaceutical compositions thereof in preparation of antibacterial drugs. The antibacterial experiment proves that the compounds C1 and C2 separated and identified by the invention have certain antibacterial effect. When the working concentration of the monomer compound is 50 mug/ml, the C1 has obvious inhibition effect on Klebsiella pneumoniae and the C1 and C2 on salmonella paratyphi B.
Detailed Description
The following description of the embodiments of the present invention is further provided in connection with the accompanying examples, and it should be noted that the embodiments described herein are for the purpose of illustration and explanation only, and are not limiting of the invention.
Example 1
Enrichment and preparation of two novel microbial secondary metabolites C1 and C2, comprising the steps of:
(1) Fermenting and producing strains: solid rice culture medium (in a 250ml conical flask, 40g rice 55ml water) is selected, and strain (Penicillium sp.) is diluted to OD 600 with sterile water, and then inoculated on the rice culture medium uniformly, and cultured for 30 days at constant temperature of 25 ℃.
(2) Preparing extract: after fermentation is completed, checking the aseptic phenomenon of a fermentation sample, mashing a fermentation product, soaking the fermentation product with ethyl acetate overnight, performing ultrasonic treatment for 1 hour, repeatedly extracting for 3-4 times, combining the extracting solutions to obtain total components, and concentrating the total components by reduced pressure distillation to obtain a total extract.
(3) Separation and purification of the compound: fermenting to obtain total extract, subjecting to silica gel column chromatography with cyclohexane-ethyl acetate system (100:1-1:100), and separating the crude fraction into three sections according to gradient elution of open glass column (column diameter 10×41cm, flow rate 6 drops/sec). The first stage is p-cyclohexane-ethyl acetate (V/V) =90: the crude fraction of 10,85:15 was purified to remove relatively less polar materials. The second stage is cyclohexane-ethyl acetate (V/V) =80: 20,70:30, combining and collecting corresponding crude fractions, taking dichloromethane-methanol (1:1) as a mobile phase (flow rate of 1 drop/second), performing gel column chromatography separation, performing ODS column chromatography separation (flow rate of 6 drops/second), and separating and purifying by using a dual-wavelength full-prepared liquid phase and methanol-water (30%) as a mobile phase (flow rate of 3 ml/min) to obtain a compound C1. The third stage is cyclohexane-ethyl acetate (V/V) =60:40, 55: 45. 50:50, using dichloromethane-methanol (1:1) as mobile phase (flow rate 6 drop/s), separating by gel column chromatography, performing ODS column chromatography (flow rate 1 drop/s), and separating and purifying by using dual-wavelength full-prepared liquid phase and methanol-water (10%) as mobile phase (flow rate 3 ml/min) to obtain compound C2.
The nuclear magnetic data of the compounds are as follows:
C1: HRESIMS showed an excimer ion peak m/z 249.0770[ M-H ] - as a yellow oil, and its molecular weight was estimated to be 250. The molecular formula was determined to be C 13H14O5 by combining 1H-NMR、13 C-NMR spectrum data, and the unsaturation degree was calculated to be 7.
1H-NMR(600MHz,CD3OD)δ:6.89(1H,d,2.0,H-2),6.69(1H,d,2.0,H-4),2.50(3H,s,5-CH3),1.40(3H,s,8-CH3),1.98(3H,s,9-CH3).13C-NMR(150MHz,CD3OD)δ:163.4(C-1),108.4(C-2),166.3(C-3),119.4(C-4),142.5(C-5),125.0(C-6),96.0(C-7),80.1(C-8),200.9(C-9),199.1(C-10),18.8(5-CH3),28.4(8-CH3),27.6(9-CH3).
C2: HRESIMS showed excimer ion m/z 141.0193[ M-H ] - as yellow oil, and its molecular weight was estimated to be 142. The molecular formula was determined to be C 6H6O4 by combining 1H-NMR、13 C-NMR data, and the unsaturation was calculated to be 4.
1H-NMR(600MHz,CD3OD)δ:2.69(2H,dt,6.3,1.5,H-3),4.45(2H,t,6.3,1.5,H-4),6.63(1H,t,1.5,H-6).13C-NMR(150MHz,CD3OD)δ:167.5(C-1),125.5(C-2),24.7(C-3),68.2(C-4),166.4(C-5),149.2(C-6).
Example 2
The antibacterial activity detection is carried out by adopting a 96-well plate turbidimetry method, the detection strains (pseudomonas aeruginosa, klebsiella pneumoniae, drug-resistant escherichia coli, candida albicans and salmonella paratyphi b) are respectively activated and detected on an LB solid culture medium, after the culture is carried out for 16 hours, a proper amount of thalli are scraped off by a sterile bamboo stick in an ultra clean bench, and sterile physiological saline is added for dilution until the OD value is measured at the wavelength of 600nm to be 0.3 (about 10 8 CFU/ml).
The bacterial suspension is diluted 10 times in an ultra clean bench, and 500 mu l to 49.5ml of liquid LB culture medium is taken to prepare uniform bacterial suspension. Test compounds (compounds C1 and C2) were diluted to 5. Mu.g/. Mu.l using molecular-grade DMSO as a mother solution for use. Soaking the 96-well plate with 75% alcohol for 30min before use, irradiating with ultraviolet for 30min in a sterile operation table, and loading into each well according to the following groups, wherein gram positive bacteria are positive control with cephalosporin and vancomycin; gram-negative bacteria have levofloxacin as a positive control; candida albicans was fluconazole as a positive control. The 96-well plate with the sample added is placed in a 37 ℃ incubator for culture, and an enzyme-labeled instrument is used for reading OD value at 600nm of ultraviolet after 18 hours. And finally, calculating the bacteriostasis rate of each sample. Antibacterial ratio (%) = (1-a/B) ×100%
A = average OD value of experimental group-OD value of blank medium control group; b = DMSO control OD.
The results of pathogen antagonism experiments (%)
1. Pseudomonas aeruginosa; 2. klebsiella pneumoniae; 3. drug resistant escherichia coli; 4. candida albicans; 5. salmonella paratyphi B; DMSO: blank control; the sample concentration was 50. Mu.g/ml.
Claims (8)
1. A microbial secondary metabolite characterized by: the secondary metabolites of the microorganism are compounds C1 and C2 shown in the following structural formula,
2. A process for the preparation of a secondary metabolite of a microorganism according to claim 1, wherein:
(1) Fermenting and producing strains:
Culturing Penicillium sp strain on rice culture medium at 26-28deg.C for 30-35 days to obtain fermented product; soaking the fermented product with ethyl acetate, performing ultrasonic treatment, and performing reduced pressure distillation to obtain crude extract, dissolving and purifying the crude extract to obtain total extract;
(2) Separation and purification of the compound:
the total extract is subjected to silica gel column chromatography by a cyclohexane-ethyl acetate system (100:1-1:100), crude fractions with cyclohexane-ethyl acetate (V/V) of 80-70:20-30 are collected, dichloromethane-methanol (1:1) is used as a mobile phase, gel column chromatography separation is carried out first, ODS column chromatography separation is carried out, and a dual-wavelength full-preparation liquid phase is used for separation and purification by using methanol-water (30%) as a mobile phase to obtain a compound C1;
Collecting cyclohexane-ethyl acetate (V/V) of 60-50:40-50, separating with dichloromethane-methanol (1:1) as mobile phase, separating with gel column chromatography, separating with ODS column chromatography, and separating and purifying with dual-wavelength full-prepared liquid phase with methanol-water (10%) as mobile phase to obtain compound C2.
3. A process for the preparation of a secondary metabolite of a microorganism according to claim 2, wherein:
and (3) dissolving the crude extract in the step (1) by using 90% methanol-water, repeatedly extracting by using cyclohexane with equal volume after dissolving, combining the extracting solutions, and concentrating under reduced pressure again to obtain the total extract.
4. A process for the preparation of a secondary metabolite of a microorganism according to claim 2, wherein:
The solid rice culture medium is prepared from 35-45g rice, 50-55mL distilled water, and is filled into 200-300mL conical flask, pH is natural, and sterilization is carried out at 121deg.C for 30-60min.
5. A process for the preparation of a secondary metabolite of a microorganism according to claim 2, wherein: the inoculation concentration of the strain (Penicillium sp.) in step (1) is: 10 5-107/mL.
6. A process for the preparation of a secondary metabolite of a microorganism according to claim 2, wherein: and (3) soaking the fermented product in the step (1) for 24-48 hours by using ethyl acetate, and performing ultrasonic treatment for 1-2 hours.
7. Use of a microbial secondary metabolite according to claim 1, characterized in that: the application of the microbial secondary metabolite in preparing a bacteriostatic agent.
8. Use of a secondary metabolite of a microorganism according to claim 7, wherein: the application of the microbial secondary metabolite in preparing an inhibitor of Klebsiella pneumoniae and salmonella paratyphi B.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211540003 | 2022-12-02 | ||
| CN2022115400035 | 2022-12-02 |
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| Publication Number | Publication Date |
|---|---|
| CN118126007A true CN118126007A (en) | 2024-06-04 |
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| CN202211571945.XA Pending CN118126007A (en) | 2022-12-02 | 2022-12-08 | Microorganism secondary metabolite and preparation method and application thereof |
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