CN101935681A - Conversion action of black rhizopus on ginsenosides - Google Patents
Conversion action of black rhizopus on ginsenosides Download PDFInfo
- Publication number
- CN101935681A CN101935681A CN2009100672173A CN200910067217A CN101935681A CN 101935681 A CN101935681 A CN 101935681A CN 2009100672173 A CN2009100672173 A CN 2009100672173A CN 200910067217 A CN200910067217 A CN 200910067217A CN 101935681 A CN101935681 A CN 101935681A
- Authority
- CN
- China
- Prior art keywords
- saponins
- ginsenosides
- panoxatriol
- bread mould
- tunning
- 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
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Steroid Compounds (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a rhizopus stolonifer capable of biologically converting ginsenosides. A fermentation product is analyzed by an HPLC (High Performance Liquid Chromatography) system to prepare a component with a reduced color spectrum peak value in an original panaxatriol saponins and a component with an increased peak value in the panaxatriol saponins fermentation product thereof. By isolation and identification, a converted substance in the panaxatriol saponins is confirmed to be ginsenosides Re, a compound (1) in the panaxatriol saponins fermentation product is confirmed to be ginsenosides Rh1, a compound (2) is confirmed to be ginsenosides Rg3, and a compound (3) is confirmed to be ginsenosides Rg2. By speculation, the ginsenosides Re in the panaxatriol saponins is converted into the ginsenosides Rh1 by the black rhizopus or the ginsenosides Re is firstly converted into the ginsenosides Rg2 and then converted into ginsenosides Rh1, and the detailed conversion path is to be further researched.
Description
Technical field:
The invention discloses a kind of bacterial classification bread mould (Rhizopus stolonifer) that makes panoxatriol saponins (ginsenosides) that bio-transformation take place, detect by the HPLC system, the result shows can make the partial monosomy saponin(e of panoxatriol saponins inside transform.Belong to microorganism in the bio-transformation Application for Field.
Background technology:
Bread mould (seeing Fig. 1 and Fig. 2) Rhizopus stolonifer is called rhizopus stolonifer again, belong to zygomycetes (Zygomycetes) mucorales (Mucorales) Zygomycotinas (Zygomycotina) fungi, be born in bread more, on the food of steamed bun and rich in starch matter, food is rotted, used bread mould is obtained by separation and purification on the wild Bulgaria inquinans infusion in Changbai Mountain among the present invention, because fungi has kind in microorganism many, secondary metabolite is many, characteristics such as culture condition is fairly simple, therefore be the major function bacterium of fermented tcm, but do not appear in the newspapers always with its research of carrying out the bio-transformation aspect of panoxatriol saponins.
Bio-transformation has excellent in chemical, zone, stereoselectivity, the reaction conditions gentleness, and the transformation efficiency height, by product is few, and product can not cause environmental pollution near natural, can carry out traditional organic synthesis and be difficult to the characteristics such as chemical reaction of carrying out.
Genseng is the dry root of Araliaceae genseng (Panax ginseng C.A.Mey.), ginsenoside is considered to the significant composition in the genseng, how to utilize biotransformation method that ginsenoside is carried out structure of modification is the focus of paying close attention to always, this research transforms the panoxatriol saponins with bread mould, and has obtained the result of study of some novelties.
Summary of the invention
The invention discloses a kind of bacterial classification bread mould (Rhizopus stolonifer) that makes panoxatriol saponins (ginsenosides) that bio-transformation take place, detect by the HPLC system, the result shows that bread mould can make the partial monosomy saponin(e of panoxatriol saponins inside transform.
The preparation of bread mould bacterial classification of the present invention can be prepared by following method:
The preparation of bread mould bacterial classification: the wild Bulgaria inquinans infusion in Changbai Mountain is placed a week, and the bread mould on the picking water liquid is connected on the PDA plating medium, pH nature, separation and purification repeatedly, up to obtaining purified bread mould bacterial classification, be transferred to PDA slant medium last 3 day, put in 4 ℃ of refrigerators standby.(bread mould is identified by professor Tu Liguer)
The preparation of ginsenoside fermented product: accurately take by weighing panoxatriol saponins 0.500 ± 0.020g and put into the 250mL triangular flask, put into 100mL PD substratum in each culturing bottle, the Autoclave of in time packing into sterilization, when vapor pressure 11.76 * 104Pa～12.75 * 104Pa, keep 30min, taking-up is placed on the Bechtop, after sterilization cools, 3 of the slant mediums of access bread mould Rhizopus stolonifer 3mm * 3mm, put shaking table into, the shaking table temperature is set at 28 ℃, revolution 125r/min, transformation time 7d final vacuum pump suction filtration fermented liquid, the saturated propyl carbinol of filtrate water extraction in 1: 1 three times is got the upper strata butanol extraction liquid and is merged, Rotary Evaporators concentrates amalgamation liquid, and concentrated solution is put evaporate to dryness obtains tunning on the water-bath.
Panoxatriol saponins fermentation production HPLC is analyzed and preparation: precision takes by weighing the tunning 20mg after concentrating, with the methyl alcohol ultrasonic dissolution and be settled in the 50mL volumetric flask, get 0.5mL and further adopt the HPLC analyte preparation, precision takes by weighing panaxsaponin mixture 20mg simultaneously, with methyl alcohol ultrasonic dissolution and constant volume in the 50mL volumetric flask, get above-mentioned solution 0.5mL and carry out HPLC and analyze, with both color atlas compare.
Table 1 panoxatriol saponins tunning liquid chromatography reaction conditions
Retention time is 120min, wavelength 203nm, 25 ℃ of column temperatures, flow velocity 2mL/min.Repeat sample introduction 5 times, circulation ratio is better.
Panoxatriol saponins fermentation production HPLC chromatogram (Fig. 4) is compared with the HPLC chromatogram (Fig. 3) of Protopanaxatriol's saponins, retention time is the compound peak value reduction of 63.244min and 101min in Protopanaxatriol's saponins HPLC collection of illustrative plates, respectively it is decided to be compd A and compd B; In the panoxatriol saponins fermentation production HPLC collection of illustrative plates, retention time is the compound 1 of 96.418min, the compound 2 that retention time is 114min, the compound 3 peak values rising that retention time is 94.319min; Prepare corresponding component, merge concentrated, standby.
The parsing of compd A:
Ginsenoside Re's standard substance HPLC collection of illustrative plates (Fig. 5) and panoxatriol saponins HPLC collection of illustrative plates (Fig. 3) are analyzed under identical conditions, infer that by retention time drawing compd A is the ginsenoside Re.
The parsing of compd B:
Panoxatriol saponins retention time is that the compd B of 101min carries out mass spectroscopy (Fig. 6), and mass spectrum [M+C1] peak is 982, and MW is 947, according to chromatographic retention and molecular weight, judges the isomers of this compound for the ginsenoside Re.
The parsing of compound 1:
With the retention time of preparing in the panoxatriol saponins tunning is that the compound 1 of 96.418min carries out mass spectroscopy (Fig. 8), and this compound mass spectrum [M-1] peak is 637, and MW is 638, in conjunction with the ginsenoside Rh
1Standard substance HPLC spectrogram (Fig. 7), according to chromatographic retention and molecular weight, authenticating compound 1 is the ginsenoside Rh
1
With the retention time of preparing in the panoxatriol saponins tunning is that the compound 2 of 114min carries out mass spectroscopy (Figure 10), and mass spectrum [M-1] peak is 783.7, and MW is 784.7, in conjunction with the ginsenoside Rg
3Standard substance HPLC spectrogram (Fig. 9) according to chromatographic retention and molecular weight, is accredited as the ginsenoside Rg
3, change into the ginsenoside Rg
3Approach await further research.
Will be from panoxatriol saponins tunning isolated retention time be that the compound 3 of 94.319min carries out mass spectroscopy (Figure 12), MW is 785, in conjunction with the ginsenoside Rg
2Standard substance HPLC spectrogram (Figure 11) is the ginsenoside Rg according to chromatographic retention and molecular weight identification
2
Inferring the bio-transformation approach (as Figure 13) of bread mould that draw to ginsenoside inside monomer saponin. the ginsenoside Re in the panoxatriol saponins is converted into the ginsenoside Rh by bread mould
1, perhaps the ginsenoside Re is converted to the ginsenoside Rg earlier
2Be converted to genseng soap Rh afterwards again
1, concrete path for transformation awaits further research.
Embodiment:
The preparation of bread mould bacterial classification: the wild Bulgaria inquinans infusion in Changbai Mountain is placed a week, and the bread mould on the picking water liquid is connected on the PDA plating medium, pH nature, separation and purification repeatedly, up to obtaining purified bread mould bacterial classification, be transferred to PDA slant medium last 3 day, put in 4 ℃ of refrigerators standby.
The preparation of ginsenoside fermented product: accurately take by weighing panoxatriol saponins 0.500 ± 0.020g and put into the 250mL triangular flask, put into the 100mLPD substratum in each culturing bottle, the Autoclave of in time packing into sterilization, when vapor pressure 11.76 * 104Pa～12.75 * 104Pa, keep 30min, taking-up is placed on the Bechtop, after sterilization cools, 3 of the slant mediums of access bread mould Rhizopus stolonifer 3mm * 3mm, put shaking table into, the shaking table temperature is set at 28 ℃, revolution 125r/min, transformation time 7d final vacuum pump suction filtration fermented liquid, the saturated propyl carbinol of filtrate water extraction in 1: 1 three times is got the upper strata butanol extraction liquid and is merged, Rotary Evaporators concentrates amalgamation liquid, and concentrated solution is put evaporate to dryness obtains tunning on the water-bath.
Panoxatriol saponins fermentation production HPLC is analyzed and preparation: precision takes by weighing the tunning 20mg after concentrating, with the methyl alcohol ultrasonic dissolution and be settled in the 50mL volumetric flask, get 0.5mL and further adopt the HPLC analyte preparation, precision takes by weighing panaxsaponin mixture 20mg simultaneously, with methyl alcohol ultrasonic dissolution and constant volume in the 50mL volumetric flask, get above-mentioned solution 0.5mL and carry out HPLC and analyze, with both color atlas compare.
Description of drawings:
Fig. 1 is the micro-synoptic diagram of bread mould.
Fig. 2 is a bread mould slant strains synoptic diagram.
Fig. 3 is a panoxatriol saponins HPLC collection of illustrative plates.
Fig. 4 is a panoxatriol saponins fermentation production HPLC collection of illustrative plates.
Fig. 5 is ginsenoside Re's standard substance HPLC collection of illustrative plates.
Fig. 6 is the compd B mass spectrum.
Fig. 7 is the ginsenoside Rh
1Standard substance HPLC collection of illustrative plates.
Fig. 8 is compound 1 mass spectrum.
Fig. 9 is the ginsenoside Rg
3Standard substance HPLC collection of illustrative plates.
Figure 10 is compound 2 mass spectrums.
Figure 11 is the ginsenoside Rg
2Standard substance HPLC collection of illustrative plates.
Figure 12 is compound 3 mass spectrums.
Figure 13 is the bio-transformation synoptic diagram of bread mould to ginsenoside.
1. ginsenoside Res, 2. ginsenoside Rg among Figure 13
2, 3. ginsenoside Rh
14. 28 ℃ of 125r/min of bread mould conversion condition.
Claims (3)
1. the preparation of bread mould bacterial classification: the wild Bulgaria inquinans infusion in Changbai Mountain is placed a week, and the bread mould on the picking water liquid is connected on the PDA plating medium, pH nature, separation and purification repeatedly, up to obtaining purified bread mould bacterial classification, be transferred to PDA slant medium last 3 day, put in 4 ℃ of refrigerators standby.(bread mould is identified by professor Tu Liguer)
The preparation of panoxatriol saponins tunning: accurately take by weighing panoxatriol saponins 0.500 ± 0.020g and put into the 250mL triangular flask, put into 100mL PD substratum in each culturing bottle, the Autoclave of in time packing into sterilization, when vapor pressure 11.76 * 104Pa～12.75 * 104Pa, keep 30min, taking-up is placed on the Bechtop, after sterilization cools, 3 of the slant mediums of access bread mould Rhizopus stolonifer 3mm * 3mm, put shaking table into, the shaking table temperature is set at 28 ℃, revolution 125r/min, transformation time 7d final vacuum pump suction filtration fermented liquid, the saturated propyl carbinol of filtrate water extraction in 1: 1 three times is got the upper strata butanol extraction liquid and is merged, Rotary Evaporators concentrates amalgamation liquid, and concentrated solution is put evaporate to dryness obtains tunning on the water-bath.
2. panoxatriol saponins and panoxatriol saponins tunning further adopt HPLC to analyze, and the contrast bread mould is to the variation on chromatographic peak before and after the fermentation of panoxatriol saponins.
3. ginsenoside tunning according to claim 2, the compound that it is characterized in that having on the said panoxatriol saponins tunning chromatographic peak compound that do not have in Protopanaxatriol's saponins and chromatogram peak value to raise occurs.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100672173A CN101935681A (en) | 2009-07-03 | 2009-07-03 | Conversion action of black rhizopus on ginsenosides |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100672173A CN101935681A (en) | 2009-07-03 | 2009-07-03 | Conversion action of black rhizopus on ginsenosides |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101935681A true CN101935681A (en) | 2011-01-05 |
Family
ID=43389250
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009100672173A Pending CN101935681A (en) | 2009-07-03 | 2009-07-03 | Conversion action of black rhizopus on ginsenosides |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101935681A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102978233A (en) * | 2012-11-16 | 2013-03-20 | 河南农业大学 | Rhizopus nigricans hypha liposome direct transformation method |
| CN104904491A (en) * | 2014-03-10 | 2015-09-16 | 廉美兰 | Method for improving saponin content in American ginseng adventive roots cultured in reactor with ginseng pathogenic bacterium elicitor |
| CN116731880A (en) * | 2023-06-16 | 2023-09-12 | 昆明理工大学 | Endophytic fungus Mucor multicastus and application thereof |
-
2009
- 2009-07-03 CN CN2009100672173A patent/CN101935681A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102978233A (en) * | 2012-11-16 | 2013-03-20 | 河南农业大学 | Rhizopus nigricans hypha liposome direct transformation method |
| CN104904491A (en) * | 2014-03-10 | 2015-09-16 | 廉美兰 | Method for improving saponin content in American ginseng adventive roots cultured in reactor with ginseng pathogenic bacterium elicitor |
| CN116731880A (en) * | 2023-06-16 | 2023-09-12 | 昆明理工大学 | Endophytic fungus Mucor multicastus and application thereof |
| CN116731880B (en) * | 2023-06-16 | 2024-04-26 | 昆明理工大学 | Endophytic fungus Mucor multicastus and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105255984A (en) | Method for converting ginsenoside through plant complex enzyme | |
| CN102086186A (en) | Method for preparing tea polyphenol rich in methylated catechin | |
| CN104530145A (en) | Method for extracting high-purity persimmon tannin from persimmon fruits | |
| CN104817610A (en) | Method for preparation of Cycloastragenol by sulfuric acid hydrolysis | |
| CN101935681A (en) | Conversion action of black rhizopus on ginsenosides | |
| CN106011213A (en) | Method for preparing cycloastragenol by bioconversion and degradation of astragaloside | |
| CN104140931A (en) | Method for preparing rare ginsenoside CK by fermenting protopanoxadiol with penicillium adametzii | |
| CN110922442B (en) | Green preparation method of ginseng extract rich in rare ginsenosides Rg3 and Rg5 | |
| CN102617674B (en) | Preparation method of scopolin monomer in anisodus tanguticus root | |
| CN104725455B (en) | Preparation method of ganoderic acid T | |
| CN103044511B (en) | A kind of isolation technique of ergot sterone in Phellinus bacterium | |
| CN100500689C (en) | Method of separating and preparing wild jujube seed saponin | |
| CN104357524A (en) | Pure wheat culture medium for promoting high yield of tritepenoidic acid from poria cocos and preparation method of pure wheat culture medium | |
| CN103044510B (en) | The isolation technique of ergosterol in Phellinus bacterium | |
| CN102643317A (en) | Method for preparing sesaminol tri-glucoside | |
| CN108383884B (en) | Separation and purification method of unstable crocin | |
| CN105063156B (en) | The new method of rubrosterone is enriched with by the modified small tuber of stemona of Fusarium oxysporum fermentation | |
| CN102060894A (en) | High-speed countercurrent chromatography for preparing isovalerylspiramycin III | |
| CN103073550B (en) | The isolation technique of ring (L-PROLINE-Valine) in Phellinus bacterium | |
| 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 | |
| CN103193748A (en) | Method for separating and preparing robinin monomer from wild chrysanthemum flowers | |
| CN102659736A (en) | Preparation method of sudachitin | |
| CN101348510B (en) | Preparation of camellia flavonol | |
| CN101633680B (en) | Process method for using ceramic membrane to replace alcohol sediment in separating puerarin |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20110105 |