CN105648002B - A method of extracting ring (sweet-junket) dipeptides from bacillus coagulans - Google Patents
A method of extracting ring (sweet-junket) dipeptides from bacillus coagulans Download PDFInfo
- Publication number
- CN105648002B CN105648002B CN201610113919.0A CN201610113919A CN105648002B CN 105648002 B CN105648002 B CN 105648002B CN 201610113919 A CN201610113919 A CN 201610113919A CN 105648002 B CN105648002 B CN 105648002B
- Authority
- CN
- China
- Prior art keywords
- dipeptides
- junket
- sweet
- ring
- volume
- 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.)
- Expired - Fee Related
Links
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
- C12P21/00—Preparation of peptides or proteins
- C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D241/00—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings
- C07D241/02—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings
- C07D241/06—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings having one or two double bonds between ring members or between ring members and non-ring members
- C07D241/08—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings having one or two double bonds between ring members or between ring members and non-ring members with oxygen atoms directly attached to ring carbon atoms
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Zoology (AREA)
- Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention discloses one kind to extract the method method of ring (sweet-junket) dipeptides from the bacillus coagulans (lactic acid bacteria).Preparing ring (sweet-junket) dipeptides method mainly includes actication of culture and culture, the acquisition of crude extract, by crude extract through silica gel column chromatography rough segmentation, gel filtration chromatography subdivision, and it is further purified through HPLC, TLC tracing detection obtains compound and detects through nuclear-magnetism and be accredited as ring (sweet-junket) dipeptides with document comparison.The raw materials used in the present invention bacillus coagulans are a kind of probiotics, and safe and harmless, extraction process is simple, at low cost；Compound purity obtained is high.
Description
Technical field
The invention belongs to the preparation technical fields of ring (sweet-junket) dipeptides, and in particular to one kind is mentioned from bacillus coagulans
The method for taking ring (sweet-junket) dipeptides.
Background technique
Bacillus coagulans (bacillus coagulans LL1103), also known as lactic acid bacteria had both possessed lactic acid bacteria production
The characteristic of lactic acid, but enzyme system and resistance abundant with bacillus are strong, high temperature resistant, bile tolerance, the characteristic easily stored.
As the up-and-coming youngster of probiotics family, be made into probiotics oneself be widely used in aquaculture, material is watched in herding,
The industries such as food and medicine.Bacillus coagulans are other than the health-care effect in maintenance intestinal microecology balances, to common
Intestines problem such as chronic colitis, diarrhea and constipation etc. have obvious curative effects, also have and adjust blood pressure and blood lipoid and improve immunity
Effect.
Research shows that having the polypeptides matter bacterium for inhibiting spoilage organisms and pathogenic bacteria in bacillus coagulans metabolite
Element generates the report of anti-plant pathogenic fungi also on bacillus coagulans in recent years.But bacillus coagulans metabolism produces
But have no that the report of small-molecule substance CYCLIC DIPEPTIDES compounds, Cyclic dipeptides are a kind of with the diketone for stablizing six-membered ring structure in object
Piperazine substance shows antibacterium, antimycotic, antiviral and antitumor, immunosupress, neuroprotection, antihyperglycemic, antimalarial
The multiple biological activities such as disease and pharmacological activity.
Summary of the invention
Ring (sweet-junket) dipeptides simple, obtained purity is high that the object of the present invention is to provide a kind of preparation processes from condensation
The method of ring (sweet-junket) dipeptides is extracted in bacillus.
To achieve the above object, the technical solution adopted by the present invention is that, one kind from bacillus coagulans extract ring (it is sweet-
Junket) dipeptides method, the structural formula of ring (sweet-junket) dipeptides is as follows:
；
It the described method comprises the following steps:
(1) actication of culture: bacillus coagulans are inoculated on plating medium, are cultivated 24 hours, are obtained under the conditions of 37 DEG C
Obtain activated spawn；
(2) primary culture: activated spawn obtained in step (1) is inoculated into MRS fluid nutrient medium, in 150r/
Min under the conditions of 37 DEG C, shaking table culture 12~16 hours, obtains seed liquor；
(3) secondary culture: the seed liquor that step (2) obtains is inoculated into MRS fluid nutrient medium, the volume of seed liquor is
The 4~6% of MRS fluid nutrient medium volume obtain fermentation liquid in shaking table culture 48~72 hours under the conditions of 150r/min, 37 DEG C；
(4) prepared by crude extract: will be centrifuged, takes after the fermentation liquid concentrated by rotary evaporation to the 1/4 of original volume of step (3) preparation
Clear liquid ethanol precipitation filters and continues to concentrate the filtrate to the 1/4 of original volume, is then extracted with ethyl acetate, by water phase
It is extracted again with n-butanol, butanol extraction liquid is concentrated under reduced pressure and obtains crude extract；
(5) silica gel column chromatography rough segmentation: filling column after the crude extract of step (4) preparation is dissolved then silica gel mixed sample with methanol,
Stationary phase is the silica gel of 80-100 mesh, with methylene chloride: methanol volume ratio=50:1~1:1 gradient elution, silica gel thin-layer chromatography inspection
It surveys, the volume ratio for merging methylene chloride-methanol is the fraction of 10:1；
(6) fraction obtained in above-mentioned steps (5) is eluted through Sephadex LH-20 gel filtration chromatography, with dichloromethane
Alkane: methanol=1:1 obtains 3 fractions as mobile phase, and through the identical fraction of silica gel thin-layer chromatography combining data detection；
(7) the third fraction reversed-phase high performance liquid chromatography for obtaining step (5) purifies, and with acetonitrile-water mobile phase, washes
De- liquid, which is concentrated under reduced pressure, removes acetonitrile and water, dry, obtains ring (sweet-junket) dipeptides.
The formula of the plating medium are as follows: peptone 10.0g/L, beef extract 10.0g/L, yeast extract 5.0g/L, grape
Sugared 20.0g/L, sodium acetate 5.0g/L, lemon acid diamine 2.0g/L, dipotassium hydrogen phosphate 0.4g/L, magnesium sulfate 0.58g/L, manganese sulfate
0.25g/L, agar 15.0g/L, Tween-80 1.0mL/L, pH 6.0.
The formula of the MRS fluid nutrient medium are as follows: peptone 10.0g/L, beef extract 10.0g/L, yeast extract 5.0g/L, Portugal
Grape sugar 20.0g/L, sodium acetate 5.0g/L, lemon acid diamine 2.0g/L, dipotassium hydrogen phosphate 0.4g/L, magnesium sulfate 0.58g/L, sulfuric acid
Manganese 0.25g/L, Tween-80 1.0mL/L, pH 6.0.
The process conditions being centrifugated after fermentation liquid concentrated by rotary evaporation in the step (4) are as follows: centrifugal condition is 4 DEG C of temperature,
Revolving speed is 5000rpm, centrifugation time 20min.
The concrete operations of step (4) the supernatant ethanol precipitation are as follows: 95% isometric second is added into supernatant
Alcohol.
The concrete operations that filtrate is extracted with ethyl acetate after concentration in the step (4) are: by filtrate after concentration and acetic acid
1 ︰ 1 is mixed ethyl ester by volume, is poured into separatory funnel, and shaking flask extracts 2 ~ 3 times, is stood 20~30min every time, is obtained water phase；
The concrete operations of the water phase that ethyl acetate is obtained by extraction extracting n-butyl alcohol are: 1 ︰ 1 is mixed water phase by volume with n-butanol,
Enter in separatory funnel, shaking flask extracts 2 ~ 3 times, stands 20~30min every time, obtains butanol extraction liquid.
Preferably, the reversed-phase high performance liquid chromatography condition in the step (7) are as follows:
Chromatographic column: Agilent pursuit C18,10 μm, 150 × 21.2mm (I.D)；
Flowing phase composition is acetonitrile: water=5: 95-50: 50；
Flow velocity: 15ml/min；
Column temperature: 25 DEG C；
Detection wavelength: 230,254nm；
Sample volume: 300-500 μ l.
The beneficial effect comprise that: the present invention carries out ring (sweet-junket) dipeptides by raw material of bacillus coagulans
It extracts, bacillus coagulans are a kind of probiotics, and metabolite has natural sex safety, and extracting method is simple, cost
It is low, there is sustainability, ring (sweet-junket) dipeptides purity is high (up to 99%) obtained, to promotion ring (sweet-junket) dipeptides pharmacology
It further studies and is of great significance as lead compound development and utilization.
Detailed description of the invention
Fig. 1 is ring (sweet-junket) dipeptides prepared by embodiment 11H NMR figure；
Fig. 2 is ring (sweet-junket) dipeptides prepared by embodiment 113C NMR figure.
Specific embodiment
The present invention is further explained in the light of specific embodiments, and but the scope of the present invention is not limited thereto.With
Bacillus coagulans used in lower embodiment from China typical culture collection center (deposit number: CCTCC NO:
M2013193).
Embodiment 1
A method of extracting ring (sweet-junket) dipeptides from bacillus coagulans, comprising the following steps:
(1) actication of culture: bacillus coagulans are inoculated on plating medium, are cultivated 24 hours, are obtained under the conditions of 37 DEG C
Obtain activated spawn；
(2) primary culture: activated spawn obtained in step (1) is inoculated into MRS fluid nutrient medium, in 150r/
Min under the conditions of 37 DEG C, shaking table culture 12 hours, obtains seed liquor；
(3) secondary culture: the seed liquor that step (2) obtains is inoculated into MRS fluid nutrient medium, the volume of seed liquor is
The 4% of MRS fluid nutrient medium volume obtains fermentation liquid in shaking table culture 48 hours under the conditions of 150r/min, 37 DEG C；
(4) prepared by crude extract: will be centrifuged, takes after the fermentation liquid concentrated by rotary evaporation to the 1/4 of original volume of step (3) preparation
Clear liquid ethanol precipitation filters and continues to concentrate the filtrate to the 1/4 of original volume, is then extracted with ethyl acetate, by water phase
It is extracted again with n-butanol, butanol extraction liquid is concentrated under reduced pressure and obtains crude extract；
(5) silica gel column chromatography rough segmentation: filling column after the crude extract of step (4) preparation is dissolved then silica gel mixed sample with methanol,
Stationary phase is the silica gel of 80-100 mesh, with methylene chloride: methanol volume ratio=50:1~1:1 gradient elution (methylene chloride: methanol
Volume ratio is 50:1,40:1,30:1,20:1,10:1,1:1), the body of methylene chloride-methanol is collected in silica gel thin-layer chromatography detection
Product is than the fraction for 10:1；
(6) fraction obtained in above-mentioned steps (5) is eluted through Sephadex LH-20 gel filtration chromatography, with dichloromethane
Alkane: methanol=1:1 obtains 3 fractions as mobile phase, and through the identical fraction of silica gel thin-layer chromatography combining data detection；
(7) the third fraction for obtaining step (5) is purified by eluant, eluent reversed-phase high performance liquid chromatography of acetonitrile-water, instead
Phase high-efficient liquid phase chromatogram condition are as follows:
Chromatographic column: Agilent pursuit C18,10 μm, 150 × 21.2mm (I.D)；
Flowing phase composition be acetonitrile: water=5: 95-50: 50(in 10min the volume ratio of acetonitrile and water gradually from 5: 95
It is transitioned into 50: 50)；
Flow velocity: 15ml/min；
Column temperature: 25 DEG C；
Detection wavelength: 230,254nm；
Sample volume: 300 μ l；
Elution time is 15min；
Eluent, which is concentrated under reduced pressure, removes acetonitrile and water, dry, obtains ring (sweet-junket) dipeptides (purity 99.1%).
The formula of the plating medium are as follows: peptone 10.0g/L, beef extract 10.0g/L, yeast extract 5.0g/L, grape
Sugared 20.0g/L, sodium acetate 5.0g/L, lemon acid diamine 2.0g/L, dipotassium hydrogen phosphate 0.4g/L, magnesium sulfate 0.58g/L, manganese sulfate
0.25g/L, agar 15.0g/L, Tween-80 1.0mL/L, pH 6.0.
The formula of the MRS fluid nutrient medium are as follows: peptone 10.0g/L, beef extract 10.0g/L, yeast extract 5.0g/L, Portugal
Grape sugar 20.0g/L, sodium acetate 5.0g/L, lemon acid diamine 2.0g/L, dipotassium hydrogen phosphate 0.4g/L, magnesium sulfate 0.58g/L, sulfuric acid
Manganese 0.25g/L, Tween-80 1.0mL/L, pH 6.0.
The process conditions being centrifugated after fermentation liquid concentrated by rotary evaporation in the step (4) are as follows: centrifugal condition is 4 DEG C of temperature,
Revolving speed is 5000rpm, centrifugation time 20min.
The concrete operations of step (4) the supernatant ethanol precipitation are as follows: 95% isometric second is added into supernatant
Alcohol.
The concrete operations that filtrate is extracted with ethyl acetate after concentration in the step (4) are: by filtrate after concentration and acetic acid
1 ︰ 1 is mixed ethyl ester by volume, is poured into separatory funnel, and shaking flask extracts 2 times, stands 30min every time, obtains water phase；Acetic acid second
The concrete operations that water phase extracting n-butyl alcohol is obtained by extraction in ester are: 1 ︰ 1 is mixed water phase by volume with n-butanol, pours into liquid separation leakage
In bucket, shaking flask is extracted 2 times, is stood 30min every time, is obtained butanol extraction liquid.
By Fig. 1-2 it is found that through1H NMR、13C NMR analysis, comparing document, (Guo Qiong, Wang Jian, Yao Junhua wait mono-
Strain South Sea coral bacterium L-4 anti-tumor activity secondary metabolite studies [J] Zhongshan University journal, 2013,52 (3):
77-82.), confirm that the compound extracted is ring (sweet-junket) dipeptides.
Above-described embodiment is the preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, it is other any without departing from the substitute mode that should be equivalent is changed made by the present invention, it is included in guarantor of the invention
Within the scope of shield.
Claims (3)
1. a kind of method for preparing ring (sweet-junket) dipeptides using microorganism, the structural formula of ring (sweet-junket) dipeptides are as follows:
；
It is characterized in that, the described method comprises the following steps:
(1) actication of culture: bacillus coagulans are inoculated on plating medium, are cultivated 24 hours, are lived under the conditions of 37 DEG C
Change strain；The deposit number of the bacillus coagulans are as follows: CCTCC NO:M2013193；
(2) primary culture: activated spawn obtained in step (1) is inoculated into MRS fluid nutrient medium, in 150r/min, 37
Under the conditions of DEG C, shaking table culture 12~16 hours, seed liquor is obtained；
(3) secondary culture: the seed liquor that step (2) obtains is inoculated into MRS fluid nutrient medium, and the volume of seed liquor is MRS
The 4~6% of fluid nutrient medium volume obtain fermentation liquid in shaking table culture 48~72 hours under the conditions of 150r/min, 37 DEG C；
(4) prepared by crude extract: will be centrifuged after the fermentation liquid concentrated by rotary evaporation to the 1/4 of original volume of step (3) preparation, takes supernatant
With ethanol precipitation, the 1/4 of original volume is filtered and continue to concentrate the filtrate to, is then extracted with ethyl acetate, by water phase with just
Butanol is extracted again, and butanol extraction liquid is concentrated under reduced pressure and obtains crude extract；
(5) silica gel column chromatography rough segmentation: filling column after the crude extract of step (4) preparation is dissolved then silica gel mixed sample with methanol, fixed
It is mutually the silica gel of 80-100 mesh, with methylene chloride: methanol volume ratio=50:1~1:1 gradient elution, silica gel thin-layer chromatography detection,
The volume ratio for collecting methylene chloride-methanol is the fraction of 10:1；
(6) fraction obtained in above-mentioned steps (5) is eluted through Sephadex LH-20 gel filtration chromatography, with methylene chloride: first
Alcohol=1:1 obtains 3 fractions as mobile phase, and through the identical fraction of silica gel thin-layer chromatography combining data detection；
(7) the third fraction for obtaining step (6) is purified using reversed-phase high performance liquid chromatography, is concentrated under reduced pressure, dry, obtains ring
(sweet-junket) dipeptides；
The formula of the plating medium are as follows: peptone 10.0g/L, beef extract 10.0g/L, yeast extract 5.0g/L, glucose
20.0g/L, sodium acetate 5.0g/L, dibasic ammonium citrate 2.0g/L, dipotassium hydrogen phosphate 0.4g/L, magnesium sulfate 0.58g/L, manganese sulfate
0.25g/L, agar 15.0g/L, Tween-80 1.0mL/L, pH 6.0；
The formula of the MRS fluid nutrient medium are as follows: peptone 10.0g/L, beef extract 10.0g/L, yeast extract 5.0g/L, glucose
20.0g/L, sodium acetate 5.0g/L, dibasic ammonium citrate 2.0g/L, dipotassium hydrogen phosphate 0.4g/L, magnesium sulfate 0.58g/L, manganese sulfate
0.25g/L, Tween-80 1.0mL/L, pH 6.0；
The concrete operations that filtrate is extracted with ethyl acetate after concentration in the step (4) are: by filtrate after concentration and ethyl acetate
1 ︰ 1 is mixed by volume, is poured into separatory funnel, and shaking flask extracts 2 ~ 3 times, is stood 20~30min every time, is obtained water phase；Acetic acid
The concrete operations that water phase extracting n-butyl alcohol is obtained by extraction in ethyl ester are: 1 ︰ 1 is mixed water phase by volume with n-butanol, pours into liquid separation
In funnel, shaking flask is extracted 2 ~ 3 times, is stood 20~30min every time, is obtained butanol extraction liquid；
Reversed-phase high performance liquid chromatography condition in the step (7) are as follows:
Chromatographic column: Agilent pursuit C18,10 μm, 150 × 21.2mm；
Flowing phase composition is acetonitrile: water=5: 95-50: 50；
Flow velocity: 15ml/min；
Column temperature: 25 DEG C；
Detection wavelength: 230,254nm；
Sample volume: 300-500 μ l.
2. the method for preparing ring (sweet-junket) dipeptides using microorganism as described in claim 1, which is characterized in that the step
(4) process conditions being centrifugated after fermentation liquid concentrated by rotary evaporation in are as follows: centrifugal condition is 4 DEG C of temperature, revolving speed 5000rpm, from
Heart time 20min.
3. the method for preparing ring (sweet-junket) dipeptides using microorganism as described in claim 1, which is characterized in that the step
(4) concrete operations of supernatant ethanol precipitation are as follows: 95% isometric ethyl alcohol is added into supernatant.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610113919.0A CN105648002B (en) | 2016-03-01 | 2016-03-01 | A method of extracting ring (sweet-junket) dipeptides from bacillus coagulans |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610113919.0A CN105648002B (en) | 2016-03-01 | 2016-03-01 | A method of extracting ring (sweet-junket) dipeptides from bacillus coagulans |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105648002A CN105648002A (en) | 2016-06-08 |
| CN105648002B true CN105648002B (en) | 2019-01-04 |
Family
ID=56492639
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610113919.0A Expired - Fee Related CN105648002B (en) | 2016-03-01 | 2016-03-01 | A method of extracting ring (sweet-junket) dipeptides from bacillus coagulans |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105648002B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102061325A (en) * | 2010-11-29 | 2011-05-18 | 大连民族学院 | Method for preparing cyclodipeptide from Bacillus amyloliquefaciens |
| CN103725738A (en) * | 2013-12-17 | 2014-04-16 | 浙江工商大学 | Method for preparing collagen polypeptides by using larimichthys crocea leftovers |
-
2016
- 2016-03-01 CN CN201610113919.0A patent/CN105648002B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102061325A (en) * | 2010-11-29 | 2011-05-18 | 大连民族学院 | Method for preparing cyclodipeptide from Bacillus amyloliquefaciens |
| CN103725738A (en) * | 2013-12-17 | 2014-04-16 | 浙江工商大学 | Method for preparing collagen polypeptides by using larimichthys crocea leftovers |
Non-Patent Citations (2)
| Title |
|---|
| 中国东海微生物F8712的分离鉴定及次生代谢产物研究;艾峰;《万方数据》;20061208;摘要，第35-52页 |
| 中国东海海洋微生物环肽类活性物质研究;朱洪平;《中国博士学位论文全文数据库 基础科学辑》;20080415(第4期);摘要，第37页第2段，第38-41页 4、实验方法，第42 -45页5、实验结果 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105648002A (en) | 2016-06-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103910708B (en) | The Azaphilones compound in a kind of thalassiomycetes source and its preparation method and application | |
| CN103937678B (en) | One strain ocean shell penicillium sp, its derivative quinolinones compound and preparation thereof and application | |
| CN102311981A (en) | Method for preparing and purifying prodigiosin | |
| CN109336873B (en) | Compound lithocarolsA-F, preparation method thereof and application thereof in preparation of antitumor drugs | |
| CN103910746A (en) | Marine fungus-derived Berkeleyones compound, and preparation method and application thereof | |
| CN106561722B (en) | The algal control purposes of thallus laminariae and the isolation and purification method of thallus laminariae algistatic activity substance | |
| CN105695544B (en) | A method of extracting ring (dried meat-is sweet) dipeptides from bacillus coagulans | |
| CN112358516B (en) | Application of diosmetin (4-O-methyl) glucoside compound in preparation of lipid-lowering drugs | |
| CN105713002B (en) | Antibiotic Versicoloids A and B and preparation method thereof and the application in anti-phytopathogen medicine is prepared | |
| CN103911407B (en) | The preparation method of the Azaphilone class dimer compound in a kind of marine fungi source and application | |
| CN103724290B (en) | Cyclopeptide compound clavatustide A as well as producing strain, preparation method and application thereof | |
| CN105648002B (en) | A method of extracting ring (sweet-junket) dipeptides from bacillus coagulans | |
| CN101481379B (en) | Compound separated from acetic acid ethyl ester extract of ginko endogenetic fungal bacterial strain fermentation liquor | |
| CN103910701A (en) | Marine fungus-derived naphthoquinone compound, and preparation method and application thereof | |
| CN105624244B (en) | A method of extracting ring (sweet-the third) dipeptides from bacillus coagulans | |
| CN105274152B (en) | Curcumin biotransformation method, product and application | |
| CN108276467A (en) | A kind of Tea Saponin and its extraction process and application | |
| CN101280333B (en) | Method for preparing penicillium antibacterial peptide from grey rose penicillium | |
| CN109234333B (en) | Method for preparing high-immunocompetence dendrobium officinale endophytic fungi polysaccharide through liquid fermentation | |
| CN104450818B (en) | A kind of method and its application that pyrroles's acid compound is prepared using hickory chick | |
| CN106565444A (en) | Extraction method and application of phenanthrene compounds from overground part of Chinese yam | |
| CN101585809B (en) | Novel compound with antibacterial and antitumor activities | |
| CN106565811B (en) | The extracting method of the hypoglycemic active ingredient of Chinese yam aerial part and application | |
| CN109988181A (en) | A kind of preparation method and application for mixing source terpene containing hydroquinone | |
| CN103525882B (en) | 9-methyl trans-streptimidone and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20190104 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |