CN110484588A - A method of Fusidic Acid is prepared using plant endogenesis epiphyte fermentation - Google Patents
A method of Fusidic Acid is prepared using plant endogenesis epiphyte fermentation Download PDFInfo
- Publication number
- CN110484588A CN110484588A CN201910842694.6A CN201910842694A CN110484588A CN 110484588 A CN110484588 A CN 110484588A CN 201910842694 A CN201910842694 A CN 201910842694A CN 110484588 A CN110484588 A CN 110484588A
- Authority
- CN
- China
- Prior art keywords
- fermentation
- fusidic acid
- preparing
- plant endogenesis
- endogenesis epiphyte
- 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
- 229960004675 fusidic acid Drugs 0.000 title claims abstract description 32
- IECPWNUMDGFDKC-MZJAQBGESA-N fusidic acid Chemical compound O[C@@H]([C@@H]12)C[C@H]3\C(=C(/CCC=C(C)C)C(O)=O)[C@@H](OC(C)=O)C[C@]3(C)[C@@]2(C)CC[C@@H]2[C@]1(C)CC[C@@H](O)[C@H]2C IECPWNUMDGFDKC-MZJAQBGESA-N 0.000 title claims abstract description 32
- IECPWNUMDGFDKC-UHFFFAOYSA-N Fusicsaeure Natural products C12C(O)CC3C(=C(CCC=C(C)C)C(O)=O)C(OC(C)=O)CC3(C)C1(C)CCC1C2(C)CCC(O)C1C IECPWNUMDGFDKC-UHFFFAOYSA-N 0.000 title claims abstract description 31
- 238000000855 fermentation Methods 0.000 title claims abstract description 23
- 230000004151 fermentation Effects 0.000 title claims abstract description 23
- 238000000034 method Methods 0.000 title claims abstract description 15
- 241000196324 Embryophyta Species 0.000 claims abstract description 11
- 239000012043 crude product Substances 0.000 claims abstract description 11
- 239000002609 medium Substances 0.000 claims abstract description 9
- 241000894006 Bacteria Species 0.000 claims abstract description 7
- 239000000284 extract Substances 0.000 claims abstract description 7
- 239000001963 growth medium Substances 0.000 claims abstract description 6
- 239000003960 organic solvent Substances 0.000 claims abstract description 6
- 241001407408 Berberis fortunei Species 0.000 claims abstract description 4
- 239000007788 liquid Substances 0.000 claims abstract description 3
- 239000000463 material Substances 0.000 claims abstract description 3
- 238000005292 vacuum distillation Methods 0.000 claims abstract description 3
- 238000011534 incubation Methods 0.000 claims abstract 2
- 238000000746 purification Methods 0.000 claims abstract 2
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 24
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 12
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 8
- 235000007164 Oryza sativa Nutrition 0.000 claims description 7
- 235000009566 rice Nutrition 0.000 claims description 7
- 239000003208 petroleum Substances 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 239000000287 crude extract Substances 0.000 claims description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 3
- 230000005526 G1 to G0 transition Effects 0.000 claims description 3
- 239000001888 Peptone Substances 0.000 claims description 3
- 108010080698 Peptones Proteins 0.000 claims description 3
- 239000012153 distilled water Substances 0.000 claims description 3
- 238000010828 elution Methods 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- 235000019319 peptone Nutrition 0.000 claims description 3
- 239000001965 potato dextrose agar Substances 0.000 claims description 3
- 238000002953 preparative HPLC Methods 0.000 claims description 3
- 239000000377 silicon dioxide Substances 0.000 claims description 3
- 239000002253 acid Substances 0.000 claims description 2
- 238000004440 column chromatography Methods 0.000 claims description 2
- 238000000605 extraction Methods 0.000 claims description 2
- 239000000741 silica gel Substances 0.000 claims description 2
- 229910002027 silica gel Inorganic materials 0.000 claims description 2
- 240000007594 Oryza sativa Species 0.000 claims 2
- 241001263206 Acremonium pilosum Species 0.000 claims 1
- 238000001228 spectrum Methods 0.000 claims 1
- 238000000926 separation method Methods 0.000 abstract description 4
- 230000001580 bacterial effect Effects 0.000 abstract description 3
- 238000002360 preparation method Methods 0.000 abstract description 3
- 230000009286 beneficial effect Effects 0.000 abstract description 2
- 241000233866 Fungi Species 0.000 abstract 1
- 241000209094 Oryza Species 0.000 description 5
- 241001019659 Acremonium <Plectosphaerellaceae> Species 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 229930188120 Carbomycin Natural products 0.000 description 1
- 238000007445 Chromatographic isolation Methods 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 206010028813 Nausea Diseases 0.000 description 1
- 206010034133 Pathogen resistance Diseases 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- 239000004098 Tetracycline Substances 0.000 description 1
- 206010047700 Vomiting Diseases 0.000 description 1
- FQVHOULQCKDUCY-OGHXVOSASA-N [(2s,3s,4r,6s)-6-[(2r,3s,4r,5r,6s)-6-[[(1s,3r,7r,8s,9s,10r,12r,14e,16s)-7-acetyloxy-8-methoxy-3,12-dimethyl-5,13-dioxo-10-(2-oxoethyl)-4,17-dioxabicyclo[14.1.0]heptadec-14-en-9-yl]oxy]-4-(dimethylamino)-5-hydroxy-2-methyloxan-3-yl]oxy-4-hydroxy-2,4-dimeth Chemical compound O([C@@H]1[C@@H](C)O[C@H]([C@@H]([C@H]1N(C)C)O)O[C@H]1[C@@H](CC=O)C[C@@H](C)C(=O)/C=C/[C@@H]2O[C@H]2C[C@@H](C)OC(=O)C[C@H]([C@@H]1OC)OC(C)=O)[C@H]1C[C@@](C)(O)[C@@H](OC(=O)CC(C)C)[C@H](C)O1 FQVHOULQCKDUCY-OGHXVOSASA-N 0.000 description 1
- 206010000059 abdominal discomfort Diseases 0.000 description 1
- 229940126575 aminoglycoside Drugs 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 229950005779 carbomycin Drugs 0.000 description 1
- 238000001460 carbon-13 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000000890 drug combination Substances 0.000 description 1
- 239000002024 ethyl acetate extract Substances 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000008693 nausea Effects 0.000 description 1
- 239000002674 ointment Substances 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 150000007660 quinolones Chemical class 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- JQXXHWHPUNPDRT-WLSIYKJHSA-N rifampicin Chemical compound O([C@](C1=O)(C)O/C=C/[C@@H]([C@H]([C@@H](OC(C)=O)[C@H](C)[C@H](O)[C@H](C)[C@@H](O)[C@@H](C)\C=C\C=C(C)/C(=O)NC=2C(O)=C3C([O-])=C4C)C)OC)C4=C1C3=C(O)C=2\C=N\N1CC[NH+](C)CC1 JQXXHWHPUNPDRT-WLSIYKJHSA-N 0.000 description 1
- 229960001225 rifampicin Drugs 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 235000019364 tetracycline Nutrition 0.000 description 1
- 150000003522 tetracyclines Chemical class 0.000 description 1
- 229940040944 tetracyclines Drugs 0.000 description 1
- 230000008673 vomiting Effects 0.000 description 1
Classifications
-
- 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
- C12P33/00—Preparation of steroids
Abstract
The invention discloses a kind of methods for preparing Fusidic Acid using plant endogenesis epiphyte fermentation, first carry out Spawn incubation to Chinese mahonia endogenetic fungus in bacterium culture medium；Carry out fermented and cultured in the fermentation medium again；Fermentation material is extracted with organic solvent, after combined extract, vacuum distillation is concentrated to get fermentation liquid medicinal extract, carries out pillar layer separation and obtains Fusidic Acid crude product；Crude product obtains Fusidic Acid sterling through chromatogram purification.The beneficial effects of the invention are as follows preparation method simplicity, and Fusidic Acid is mass produced using the bacterial strain.
Description
Technical field
The invention belongs to technical field of chemistry, are related to a kind of side that Fusidic Acid is prepared using plant endogenesis epiphyte fermentation
Method.
Background technique
Fusidic Acid is a kind of important clinical antibacterials, clinically generally using Fusidic Acid sodium salt (injection or
Ointment), there is extremely strong penetration power to tissue and body fluid, plasma half-life is long when intravenously administrable, and patient tolerability is good.Drug resistance
Property aspect: without cross resistance between Fusidic Acid and other antibacterials of clinical use, there is good safety.Fu Xi
Ground acid can often be used with Drug combinations, collaborations such as rifampin, carbomycin, aminoglycoside, quinolones, Tetracyclines
Medicine significant effect.In terms of side effect: patient is general, and tolerance is good, and major side effects have gastrointestinal discomfort, nausea, digestion occur
Bad, vomiting etc..Country's report developed with separation purifying technique that ferments about Fusidic Acid is less.Country's Fusidic Acid at present
Production technology yield is relatively low, and clinic most of Fusidic Acid drug in China's uses foreign import, and expensive.Therefore,
A kind of method simplicity is developed, the Fusidic Acid large-scale producing method that raw material is easy to get is particularly important.
Summary of the invention
The purpose of the present invention is to provide a kind of method for preparing Fusidic Acid using plant endogenesis epiphyte fermentation, the present invention
Beneficial effect be preparation method simplicity, using the bacterial strain be mass produced Fusidic Acid.
The technical scheme adopted by the invention is that following the steps below:
(1) strain training first is carried out to Chinese mahonia endogenetic fungus Acremonium pilosumF47 in bacterium culture medium
It supports；
(2) fermented and cultured is carried out in the fermentation medium again；
(3) fermentation material to be extracted with organic solvent, after combined extract, vacuum distillation is concentrated to get fermentation liquid medicinal extract, into
Row pillar layer separation obtains crude product；
(4) above-mentioned crude product obtains Fusidic Acid sterling, for purity 98% or more, yield is up to 87.5mg/ through chromatographic isolation
kg。
Further, bacterium culture medium is potato dextrose agar in step (1), is cultivated 3 days.
Further, step (2) fermentation medium: rice medium -80g rice, 120ml distilled water, 0.3% peptone,
500ml conical flask, 28 DEG C constant temperature stationary culture 40-50 days.
Further, organic solvent uses ethyl acetate in step (3).
Further, crude extract is crossed silica normal phase column chromatography by extraction process in step (3), and stationary phase is 200-300 mesh silicon
Glue, petroleum ether and ethyl acetate mixture are mobile phase, and the isolated Fusidic Acid mixture of gradient elution collects petroleum ether:
Ethyl acetate=2:1~1:2 component, is concentrated to give crude product.
Further, crude product is purified through preparative high performance liquid chromatography in step (4), and first alcohol and water is mobile phase, obtains husband
The western sour sterling in ground.
Detailed description of the invention
Fig. 1 is Fusidic Acid sterling chemical formula.
Specific embodiment
The present invention is described in detail With reference to embodiment.
Embodiment: endogenetic fungal bacterial strain: Acremonium pilosumF47；Bacterium source: it is big to be isolated from plant broad-leaved ten
Contribution floral axis；Bacterium culture medium: potato dextrose agar；Fermentation medium: rice medium -80g rice,
120ml distilled water, 0.3% peptone, 500ml conical flask, 28 DEG C constant temperature stationary culture 40-50 days；It extracts: using organic solvent
Ethyl acetate extracts, and obtains crude extract；Separation: column chromatographic runs, crude extract cross silica normal phase column, and stationary phase is 200-300 mesh
Silica gel, petroleum ether and ethyl acetate be mobile phase, gradient elution (100:0-0:100, v/v), collect petroleum ether: ethyl acetate=
2:1~1:2 component obtains Fusidic Acid crude product；Purifying: obtained Fusidic Acid crude product is pure through preparative high performance liquid chromatography
Change, first alcohol and water is mobile phase, obtains Fusidic Acid sterling.
Yield: about 87.5mg Fusidic Acid/1000g rice, 98.5% or more purity.
Fusidic Acid sterling chemical formula is as shown in Figure 1.Structural identification data is as shown in table 1.
1 carbon-13 nmr spectra of table and hydrogen modal data (400,100MHz, CDCl3,TMS,δppm)
The present invention using Chinese mahonia endogenetic fungus Acremonium pilosumF47 as engineered strain, rapid, high volume
The higher Fusidic Acid of purity is prepared, and preparation process is simple, efficiently.
The above is only not to make limit in any form to the present invention to better embodiment of the invention
System, any simple modification that embodiment of above is made according to the technical essence of the invention, equivalent variations and modification,
Belong in the range of technical solution of the present invention.
Claims (6)
1. a kind of method for preparing Fusidic Acid using plant endogenesis epiphyte fermentation, it is characterised in that:
(1) Spawn incubation first is carried out to Chinese mahonia endogenetic fungus Acremonium pilosum F47 in bacterium culture medium；
(2) fermented and cultured is carried out in the fermentation medium again；
(3) fermentation material is extracted with organic solvent, after combined extract, vacuum distillation is concentrated to get fermentation liquid medicinal extract, carries out color
Spectrum separates to obtain crude product；
(4) crude product obtains Fusidic Acid sterling, for purity 98% or more, yield is up to 87.5mg/kg through chromatogram purification.
2. according to a kind of method for preparing Fusidic Acid using plant endogenesis epiphyte fermentation described in claim 1, it is characterised in that:
Bacterium culture medium is potato dextrose agar in the step (1), is cultivated 3 days.
3. according to a kind of method for preparing Fusidic Acid using plant endogenesis epiphyte fermentation described in claim 1, it is characterised in that:
Step (2) fermentation medium: rice medium -80g rice, 120ml distilled water, 0.3% peptone, 500ml taper
Bottle, 28 DEG C constant temperature stationary culture 40-50 days.
4. according to a kind of method for preparing Fusidic Acid using plant endogenesis epiphyte fermentation described in claim 1, it is characterised in that:
Organic solvent uses ethyl acetate in the step (3).
5. according to a kind of method for preparing Fusidic Acid using plant endogenesis epiphyte fermentation described in claim 1, it is characterised in that:
Crude extract is crossed silica normal phase column chromatography by extraction process in the step (3), and stationary phase is 200-300 mesh silica gel, petroleum ether and
Ethyl acetate mixture is mobile phase, and gradient elution collects petroleum ether: ethyl acetate=2:1~1:2 component, isolated husband
Western ground acid crude.
6. according to a kind of method for preparing Fusidic Acid using plant endogenesis epiphyte fermentation described in claim 1, it is characterised in that:
Obtained crude product is purified through preparative high performance liquid chromatography in the step (4), first alcohol and water is mobile phase, obtains Fu Xidi
Sour sterling.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910842694.6A CN110484588A (en) | 2019-09-06 | 2019-09-06 | A method of Fusidic Acid is prepared using plant endogenesis epiphyte fermentation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910842694.6A CN110484588A (en) | 2019-09-06 | 2019-09-06 | A method of Fusidic Acid is prepared using plant endogenesis epiphyte fermentation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110484588A true CN110484588A (en) | 2019-11-22 |
Family
ID=68556949
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910842694.6A Pending CN110484588A (en) | 2019-09-06 | 2019-09-06 | A method of Fusidic Acid is prepared using plant endogenesis epiphyte fermentation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110484588A (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080318267A1 (en) * | 2004-12-17 | 2008-12-25 | Cubist Pharmaceuticals, Inc. | Methods and Compositions for Ultra-High Throughput Screening of Natural Products |
| US20130266645A1 (en) * | 2010-10-06 | 2013-10-10 | Justus-Liebig-Universitat Giessen | Derivatives of steroid benzylamines, having an antiparasitic antibacterial, antimycotic and/or antiviral action |
| US20160338360A1 (en) * | 2013-12-24 | 2016-11-24 | Indigo Ag, Inc. | Plants containing beneficial endophytes |
-
2019
- 2019-09-06 CN CN201910842694.6A patent/CN110484588A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080318267A1 (en) * | 2004-12-17 | 2008-12-25 | Cubist Pharmaceuticals, Inc. | Methods and Compositions for Ultra-High Throughput Screening of Natural Products |
| US20130266645A1 (en) * | 2010-10-06 | 2013-10-10 | Justus-Liebig-Universitat Giessen | Derivatives of steroid benzylamines, having an antiparasitic antibacterial, antimycotic and/or antiviral action |
| US20160338360A1 (en) * | 2013-12-24 | 2016-11-24 | Indigo Ag, Inc. | Plants containing beneficial endophytes |
Non-Patent Citations (2)
| Title |
|---|
| 程洁等: "顶孢霉发酵液提取物清除自由基能力的研究", 《食品工业科技》 * |
| 钱思宇等: "夫西地酸发酵培养基优化", 《河北化工》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104694401A (en) | Penicillium verrucosum LPPV001 and method for producing ergosterol peroxide by same | |
| CN102061320A (en) | Preparation method of 11 alpha,17 alpha-dyhydroxyl-androst-4-ene-3,20-dione | |
| CN109134574B (en) | Steroid compound, preparation method and application thereof, and anti-tumor drug | |
| CN104276956A (en) | Method for preparing S-1-tetralin amine | |
| CN110484588A (en) | A method of Fusidic Acid is prepared using plant endogenesis epiphyte fermentation | |
| CN109136111B (en) | Method for large-scale preparation of tryptophan-alanine cyclic dipeptide by using marine fungi | |
| CN109971651B (en) | Tobacco endophytic fungus and application thereof in preparation of ergosterol 5,8 peroxide | |
| CN109053638A (en) | A kind of method for extraction and purification of fumidil | |
| CN103214543B (en) | New Crategolic acid derivative, its preparation method and the application in antitumor drug thereof | |
| CN102296039A (en) | Pseudonocardia and method for preparing Deoxynyboquinone by same | |
| CN107354182B (en) | A kind of method that ash green soy bean endogenetic fungus fermentation prepares (R) -4- benzyl -2- oxazolidinone compounds | |
| CN108727169A (en) | A kind of preparation method of the biphenyl ether compound in marine fungi source and the application as antiseptic | |
| CN108774120B (en) | Lapatiquinone compound and preparation method thereof | |
| CN103044510A (en) | Technology for separating ergosterol from Phellinus | |
| CN115976093B (en) | Method for preparing orcein by using aspergillus oryzae | |
| CN103130809B (en) | The isolation technique of six hydrogen-7-hydroxyl-3-(2-methyl-propyl) pyrrolo-[1,2-a] pyrazine-Isosorbide-5-Nitrae-diketone in Phellinus bacterium | |
| CN108658911B (en) | Furanonaquinone compound and preparation method thereof | |
| CN108586435A (en) | The preparation method of Echinulin | |
| CN103073550B (en) | The isolation technique of ring (L-PROLINE-Valine) in Phellinus bacterium | |
| CN106011190B (en) | Preparation method of natural p-aminobenzoic acid and derivatives thereof | |
| CN102391145B (en) | Method for resolving optical isomer of ubenimex intermediate conveniently | |
| CN110218750B (en) | Application of aspergillus versicolor strain in fermentation production of kojic acid | |
| CN108929857B (en) | Salicornia europaea-derived marine fungus and application thereof in preparation of topoisomerase I inhibitor | |
| CN104630304B (en) | A kind of method that utilization microorganism prepares sophoridine oxide | |
| CN103044376A (en) | Technology for separating pyrone from Phellinus |
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 | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20191122 |
|
| RJ01 | Rejection of invention patent application after publication |