CN109082445A - The metabolite product of one plant of gingko endogenous fungus and its application in antibacterial - Google Patents
The metabolite product of one plant of gingko endogenous fungus and its application in antibacterial Download PDFInfo
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Abstract
This disclosure relates to the metabolite product of one plant of gingko endogenous fungus and its application in antibacterial, fungi used is the raw sickle-like bacteria of gingko endogenous fungus layer (Fusarium proliferatum) DZHQ1, deposit number is CGMCC No.14983, China Committee for Culture Collection of Microorganisms's common micro-organisms center is preserved on November 28th, 2017, fermented culture and extraction separation obtain the metabolite of gingko endogenous fungus.The metabolite of the gingko endogenous fungus of the disclosure shows obvious inhibiting effect in anti-Escherichia coli and/or staphylococcus aureus Activity determination, has the potential use for preparing new antimicrobial product.
Description
Technical field
The disclosure belongs to microbial medicine field, and in particular to the metabolite product of one plant of gingko endogenous fungus and its
Application in antibacterial.
Background technique
Here statement only provides background information related with the disclosure, without necessarily constituting the prior art.
Escherichia coli are typical gram-Negative bacillus, and caused colibacillosis is a kind of common disease, beast
It is extremely serious to cure harm caused by clinically Escherichia coli, can cause a disease throughout the year, is always the normal of puzzlement aquaculture development
See disease, frequently-occurring disease, causes serious economic loss to aviculture.It is easy to produce drug resistance, and Antibiotic Resistance over the course for the treatment of
Extensively, resistance mechanism is complicated, prevents and treats the disease to aviculture and brings very big difficulty.Therefore a kind of confrontation Escherichia coli are found
Effective ingredient becomes research hotspot in recent years.
Staphylococcus aureus is one of inside and outside most common pathogen of infection of hospital, and Chinese Bacterial resistance surveillance net is more
Over year 5 before the clinically isolated bacterium bacterial strain of the tens of institute's infection from hospital in the whole nation shows that staphylococcus aureus accounts for clinical bacteria
Position.The bacterium can cause serious pneumonia, meningitis and bone joint infection, endocarditis and bacteremia.Especially burn wound
The Diseases easy infection such as infection, the acute liver failure and haematogenous ephritis bacterium.In recent years, some drug susceptibilities are dropped
The appearance of low staphylococcus aureus will make facing mankind is certain to infect the situation to past medical help.Therefore, a kind of confrontation is found
The effective ingredient of staphylococcus aureus is of great significance.
CN108102928A " one plant of gingko endogenous fungus and its application " discloses one plant and is isolated to from ginkgo bark
With the active raw sickle-like bacteria of gingko endogenous fungus layer (Fusarium proliferatum) DZHQ1 of anti-cervical cancer, deposit number
For CGMCC No.14983, which also discloses the fermentation cultures of the endogenetic fungus, the ethyl acetate extract of fermentation culture
There is therapeutic activity for cervical carcinoma.
Further carry out Gingko biloba plant endogenetic fungus active matter Quality Research, to the exploitation medicinal Nei Shengzhen of China's Gingko biloba plant
Bacterium resource and new microbial medicine all have important theory significance and potential application value.
Summary of the invention
For background technique, the disclosure further grinds CN108102928A " one plant of gingko endogenous fungus and its application "
Study carefully and explore, provide the metabolite product of one plant of gingko endogenous fungus and its is preparing the application in antimicrobial product.
The disclosure specifically uses following technical scheme:
In the first aspect of the disclosure, the metabolite product of one plant of gingko endogenous fungus is provided, its main feature is that: institute
Fungi is the raw sickle-like bacteria of gingko endogenous fungus layer (Fusarium proliferatum) DZHQ1, deposit number CGMCC
No.14983 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on November 28th, 2017,
Fermented culture and ethyl acetate obtain the ethyl acetate extract of fermentation culture after extracting, then using methanol or water-soluble
Solution obtains the metabolite product of gingko endogenous fungus after dissolution.
In the second aspect of the disclosure, the preparation method of the metabolite product of one plant of gingko endogenous fungus is provided,
This method comprises: fungi used is the raw sickle-like bacteria of gingko endogenous fungus layer (Fusarium proliferatum) DZHQ1, protect
Hiding number is CGMCC No.14983, and it is general to be preserved in China Committee for Culture Collection of Microorganisms on November 28th, 2017
The ethyl acetate extract for the fermentation culture that logical microorganism center, fermented culture and ethyl acetate obtain after extracting, then
It is dissolved using methanol or water, dissolved lysate is the metabolite product of gingko endogenous fungus.
In terms of the third of the disclosure, the metabolite product for providing the gingko endogenous fungus is produced in preparation antibacterial
Application in product.
At the 4th aspect of the disclosure, a kind of bactericidal composition is provided, with the metabolism of the gingko endogenous fungus
Product is active constituent.
At the 5th aspect of the disclosure, a kind of antibiotic method is provided, this method includes using the gingko endogenous fungus
Metabolite product or the bactericidal composition inhibit bacterium the step of.
The one of technical solution of the disclosure has the following beneficial effects:
The metabolite of the gingko endogenous fungus of the disclosure is shown in anti-Escherichia coli and/or staphylococcus aureus detection
Obvious inhibiting effect is shown, there is the potential use for preparing new antimicrobial product.
Detailed description of the invention
The Figure of description for constituting disclosure a part is used to provide further understanding of the disclosure, the signal of the disclosure
Property embodiment and its explanation for explaining the disclosure, do not constitute the improper restriction to the disclosure.
Fig. 1: 17#The cation chromatogram of (17-1 and 17-2) sample.
Fig. 2: 18#The cation chromatogram of (18-1 and 18-2) sample.
Fig. 3: 17#The anion chromatogram of (17-1 and 17-2) sample.
Fig. 4: 18#The anion chromatogram of (18-1 and 18-2) sample.
Fig. 5: 17#With 18#Inhibitory effect of the gingko endogenous fungus crude extract methanol solution to Escherichia coli.
Fig. 6: 17#With 18#Inhibitory effect of the gingko endogenous fungus crude extract aqueous solution to Escherichia coli.
Fig. 7: 17#With 18#Inhibitory effect of the gingko endogenous fungus crude extract methanol solution to staphylococcus aureus.
Fig. 8: 17#With 18#Inhibitory effect of the gingko endogenous fungus crude extract aqueous solution to staphylococcus aureus.
Specific embodiment
It is noted that described further below be all exemplary, it is intended to provide further instruction to the disclosure.Unless another
It indicates, all technical and scientific terms used herein has usual with disclosure person of an ordinary skill in the technical field
The identical meanings of understanding.
It should be noted that term used herein above is merely to describe specific embodiment, and be not intended to restricted root
According to the illustrative embodiments of the disclosure.As used herein, unless the context clearly indicates otherwise, otherwise singular
Also it is intended to include plural form, additionally, it should be understood that, when in the present specification using term "comprising" and/or " packet
Include " when, indicate existing characteristics, step, operation and/or their combination.
The applicant makees into one research achievement CN108102928A " one plant of gingko endogenous fungus and its application " before
Step research and exploration, propose the metabolite product of one plant of gingko endogenous fungus, its main feature is that: fungi used is
The raw sickle-like bacteria of gingko endogenous fungus layer (Fusarium proliferatum) DZHQ1, deposit number are in CN108102928A
CGMCC No.14983 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms on November 28th, 2017
Center, fermented culture and ethyl acetate obtain the ethyl acetate extract of fermentation culture after extracting, then using methanol
Or water dissolution, dissolved lysate is the metabolite product of gingko endogenous fungus.
In one of the disclosure or some specific embodiments, the side of fermented and cultured and ethyl acetate extraction is provided
Method, comprising: take cultured gingko endogenous fungus in PDA solid medium, by inoculated by hypha block in PDA liquid medium
In, 20~28 DEG C, 100~150r/min shaking table culture, 5~7d；Then ethyl acetate (second is added by the volume ratio of 1:1~2:1
Acetoacetic ester and fermentating liquid volume ratio are 1:1-2:1), continue to cultivate 4~8d in shaking table；Fermentation liquid removes mycelium after filtering,
It is layered again, obtains the organic phase containing gingko endogenous fungus secondary metabolite；Ethyl acetate is recycled, is obtained containing ginkgo
The concentrate of endogenetic fungus secondary metabolite；Finally, concentrate is dry, as the ethyl acetate of fermentation culture is extracted
Object.
Wherein, PDA solid medium and PDA liquid medium are potato culture conventional in the prior art, formula
It may be selected but be not limited solely to: potato 200g, glucose 20g, 15~20g of agar and water 1000mL；Alternatively, potato
200g, glucose 20g and water 1000mL.
In one of the disclosure or some specific embodiments, the volume ratio of fermentation liquid and ethyl acetate is 1:1
~1:2.
In a specific embodiment of the disclosure, the metabolite product of the gingko endogenous fungus activity at
Point include but be not restricted to glycine betaine, Scopoletin, yageine, Rosmarinic acid, oxipurinol, resveratrol,
Naringenin, catechin, texifolin and xanthohumol.
Further, the metabolite product active ingredient of gingko endogenous fungus obtained after methanol dissolution includes but not
It is only limited to glycine betaine, Scopoletin, yageine, Rosmarinic acid, oxipurinol, resveratrol, naringenin, catechu
Element, texifolin and xanthohumol.
In a specific embodiment of the disclosure, glycine betaine, Scopoletin, yageine, Rosmarinic acid,
Oxipurinol, resveratrol, naringenin, catechin, texifolin and xanthohumol are peak area in HPLC figure 105Above peak
The substance of representative.
In a specific embodiment of the disclosure, the metabolite of the gingko endogenous fungus for example table 3 the 6th column or
Shown in 7th column.
In one of the disclosure or some specific embodiments, the ratio of ethyl acetate extract and methanol or water is
(0.01~0.05) g:1mL；Further ratio is 0.02g:1mL.
In a typical embodiment of the disclosure, the metabolite product of one plant of gingko endogenous fungus is provided
Preparation method, this method comprises: fungi used is the raw sickle-like bacteria of gingko endogenous fungus layer (Fusarium proliferatum)
DZHQ1, deposit number are CGMCC No.14983, are preserved in Chinese microorganism strain preservation management on November 28th, 2017
Committee's common micro-organisms center, fermented culture and ethyl acetate obtain fermentation culture ethyl acetate after extracting extracts
Object obtains methanol or the dissolved gingko endogenous fungus of water then using methanol or water dissolution (5~30min) after dissolution
Metabolite product.
In a specific embodiment of the disclosure, filter membrane after dissolution；Further, the aperture of the filter membrane is
0.22μm。
In another typical embodiment of the disclosure, the metabolite product for providing the gingko endogenous fungus exists
Prepare the application in antimicrobial product.
In a specific embodiment of the disclosure, the antimicrobial product is antibacterials or other preparations.
In a specific embodiment of the disclosure, the metabolite product of the gingko endogenous fungus can resist greatly
Enterobacteria and/or staphylococcus aureus.
In a specific embodiment of the disclosure, the metabolite of the dissolved gingko endogenous fungus of methanol
Product can resist Escherichia coli.
In a specific embodiment of the disclosure, the metabolism of the methanol or the dissolved gingko endogenous fungus of water
Product can resist Escherichia coli/or staphylococcus aureus.
In a typical embodiment of the disclosure, a kind of bactericidal composition is provided, with described gingko endogenous
The metabolite product of fungi is active constituent.
In a specific embodiment of the disclosure, the composition also includes pharmaceutically acceptable carrier, is carried
Body be selected from one or more of diluent, dispersing agent, stabilizer, disintegrating agent, lubricant, as starch, sodium carboxymethylcellulose,
Glycerol etc. is one such or a variety of.
In a typical embodiment of the disclosure, a kind of antibiotic method is provided, this method includes described in use
The step of metabolite product of gingko endogenous fungus or the bactericidal composition inhibit bacterium, wherein the bacterium is large intestine
Bacillus and/or staphylococcus aureus.
In order to enable those skilled in the art can clearly understand the technical solution of the disclosure, below with reference to tool
The technical solution of the disclosure is described in detail in the embodiment of body.
Embodiment 1
A kind of preparation method of the ethyl acetate extract of fermentation culture can refer to CN108102928A " one plant of ginkgo
Endogenetic fungus and its application " embodiment 2 in content.
Specifically, mycelia block is connect this method comprises: take cultured gingko endogenous fungus in PDA solid medium
Kind is in the triangular flask equipped with 250ml PDA liquid medium, 20 DEG C, 120r/min shaking table culture 7d.Then to each culture
Ethyl acetate is added by the volume ratio of 1:1 in bottle, 4d is cultivated in continuation in shaking table.Fermentation liquid degerming after 3-4 layers of filtered through gauze
Filament is layered with separatory funnel, obtains the organic phase containing gingko endogenous fungus secondary metabolite.Utilize rotary evaporation
Instrument recycles ethyl acetate most of in organic phase, obtains the concentrate containing gingko endogenous fungus secondary metabolite.Finally,
Concentrate is completely dried using vacuum concentration drier, as the ethyl acetate extract of fermentation culture.Wherein, the silver
Apricot endogenetic fungus is the raw sickle-like bacteria of gingko endogenous fungus layer (Fusarium proliferatum) DZHQ1, and deposit number is
CGMCC No.14983, number 17#J-1 bacterial strain in corresponding CN108102928A " one plant of gingko endogenous fungus and its application ".
Alternatively, the gingko endogenous fungus is the J-2 bacterial strain in CN108102928A " one plant of gingko endogenous fungus and its application ", number
18#。
The preparation method of the metabolite product of one plant of gingko endogenous fungus, the ethyl acetate of the fermentation culture is mentioned
Object is taken to be dissolved using methanol, the ethyl acetate extract of the fermentation culture and the ratio of methanol are 0.02g:1mL, molten
The lysate obtained after solution 10min is the metabolite product of gingko endogenous fungus.
Embodiment 2
A kind of preparation method of the metabolite product of gingko endogenous fungus, by fermentation culture described in embodiment 1
Ethyl acetate extract is dissolved using water, and the ethyl acetate extract of the fermentation culture and the ratio of water are 0.02g/
mL.Wherein, the gingko endogenous fungus is the raw sickle-like bacteria of gingko endogenous fungus layer (Fusarium proliferatum) DZHQ1,
Deposit number is CGMCC No.14983, number 17#In corresponding CN108102928A " one plant of gingko endogenous fungus and its application "
J-1 bacterial strain.
Embodiment 3
The preparation method of the metabolite product of one plant of gingko endogenous fungus, by the second of fermentation culture described in embodiment 1
Acetoacetic ester extract is dissolved using methanol or water, the ethyl acetate extract and methanol of the fermentation culture or the ratio of water
Example is 0.02g/mL, crosses 0.22 μm of filter membrane after dissolving 10min, obtained filtrate is that the metabolite of gingko endogenous fungus produces
Product.Wherein, the gingko endogenous fungus is the raw sickle-like bacteria of gingko endogenous fungus layer (Fusarium proliferatum) DZHQ1,
Deposit number is CGMCC No.14983, number 17#In corresponding CN108102928A " one plant of gingko endogenous fungus and its application "
J-1 bacterial strain.Alternatively, the gingko endogenous fungus is the J- in CN108102928A " one plant of gingko endogenous fungus and its application "
2 bacterial strains, number 18#.Finally obtain 17#Sample, it may be assumed that the metabolite product or water of the dissolved gingko endogenous fungus of methanol
The metabolite product of dissolved gingko endogenous fungus；18#Sample, it may be assumed that the metabolism of the dissolved gingko endogenous fungus of methanol
The metabolite product of product or the dissolved gingko endogenous fungus of water.
Embodiment 4
The preparation method of the metabolite product of one plant of gingko endogenous fungus, this method comprises: taking PDA solid medium
In the cultured raw sickle-like bacteria of gingko endogenous fungus layer (Fusarium proliferatum) DZHQ1, by inoculated by hypha block in dress
In the triangular flask for having 250ml PDA liquid medium, 25 DEG C, 100r/min shaking table culture 6d.Then it is pressed into each culture bottle
Ethyl acetate is added in the volume ratio of 2:1, and 3d is cultivated in continuation in shaking table.Fermentation liquid removes mycelium after 3-4 layers of filtered through gauze, uses
Separatory funnel is layered, and the organic phase containing gingko endogenous fungus secondary metabolite is obtained.To have using Rotary Evaporators
Most of ethyl acetate recycling, obtain the concentrate containing gingko endogenous fungus secondary metabolite in machine phase.Finally, using true
Sky is concentrated and dried device and is completely dried concentrate, as the ethyl acetate extract of fermentation culture, by the fermentation culture
Ethyl acetate extract dissolved using methanol or water, the ethyl acetate extract and methanol or water of the fermentation culture
Ratio be 0.02g/mL, dissolve the metabolite product that obtained lysate after 10min is gingko endogenous fungus.
Embodiment 5
The preparation method of the metabolite product of one plant of gingko endogenous fungus, this method comprises:
Take cultured three plants of gingko endogenous fungus in PDA solid medium, by inoculated by hypha block in equipped with 250ml
In the triangular flask of PDA liquid medium, 28 DEG C, 140r/min shaking table culture 8d.Then the body of 1.5:1 is pressed into each culture bottle
Product cultivates 5d than ethyl acetate, continuation is added in shaking table.Fermentation liquid removes mycelium after 3-4 layers of filtered through gauze, is leaked with liquid separation
Bucket is layered, and the organic phase containing gingko endogenous fungus secondary metabolite is obtained.It will be in organic phase using Rotary Evaporators
Most of ethyl acetate recycling, obtain the concentrate containing gingko endogenous fungus secondary metabolite.Finally, utilizing vacuum concentration
Concentrate is completely dried by drier, and the ethyl acetate extract of the fermentation culture is dissolved using methanol or water,
The ethyl acetate extract and methanol of the fermentation culture or the ratio of water are 0.02g/mL, are obtained after dissolution 10min molten
Solution liquid is the metabolite product of gingko endogenous fungus.
The non-targeted metabolism group detection of 6 LC-MS of embodiment
1.1 material
1.1.1 sample essential information
Totally 2 group (17 of sample (ethyl acetate extract of the fermentation culture in embodiment 1)#,18#), every group 2 are parallel,
Amount to 4 samples (17-1,17-2,18-1,18-2).
1.1.2 reagent
Water (water, Watson), reagent: mass spectrum acetonitrile (Fisher), formic acid (Sigma-Aldrich)
1.1.3 instrument
Device information table 1
1.2 method
1.2.1 metabolin extracts
1mL methanol is added into sample and dissolves (sample: methanol=0.02g:1mL) 10min, it is laggard to cross 0.22 μm of filter membrane
Sample.
1.2.2 LC-MS analysis condition
The analysis instrument of this experiment is Q Exactive plus, Thermo, ion source ESI.
1. chromatographic condition
Chromatographic column is Waters Atlantis T3 (100 × 3mm, 1.8mm).Column temperature is 35 DEG C.Flow velocity is 0.500 [ml/
min]。
Mobile phase: A.Equate=" 0.1v/v%HCOOH-H2O"
D.Equate=" acetonitrile "
Chromatographic condition see the table below: table 2
2. Mass Spectrometry Conditions
Scanning range selects m/z 80-1200；
Resolution ratio: 70,000；
Spectrum data type:Profile；
3500 V (negative) of Capillary voltage:4000V (positive) and；
Capillary Temp:350℃.
2 results and analysis
2.1 cation chromatograms
Such as Fig. 1 and 2.
2.2 anion chromatograms
Such as Fig. 3 and 4.
2.3 identification
The initial data of LC-MS is imported into (the MS-DIAL:data independent MS/MS of MS-DIAL 2.76
deconvolution for comprehensive metabolome analysis.Nature Methods,12,523-
526,2015) software is pre-processed, including peak extracts, removes noise, deconvolution, and peak alignment exports the three-dimensional data of CSV format
Matrix (raw data matrix).The peak information and date library of extraction is compared, to MassBank, Respect, GNPS are (altogether
14951 records) three libraries carry out full library searching.The information that this three-dimensional matrice includes has: sample message, retention time,
Karyoplasmic ratio and mass spectrum response intensity (peak area).
2.4 interpretation of result
Qualification result such as the following table 3.
Table 3
The substance that this not good liquor matter identifies sees the above table 3, and wherein inventor is by investigation, the main active component having been reported
(main active component refers to peak area 105The substance that above peak represents), 17#It include: glycine betaine, Scopoletin, dehydrogenation camel
Fluffy alkali, Rosmarinic acid, oxipurinol, resveratrol, naringenin, Kaempferol, catechin, texifolin and xanthohumol；18#Packet
It includes: Scopoletin, yageine, Rosmarinic acid, oxipurinol, resveratrol, naringenin, Kaempferol, catechin, star-spangled banner
Song Su and xanthohumol.
Since liquid quality detection can only detect the compound being currently known, so the matter interpretation of result of this not good liquor has gone out part and has had
Antitumor and antibacterial activity component, therefore for whether also needing containing unknown reactive compound into one in fermentation liquid
Step research.
The research of 5 In Vitro Bacteriostasis of embodiment
1, experimental procedure:
Detected bacterium is inoculated into the triangular flask equipped with 100ml LB liquid medium, at 37 DEG C, 120r/min shaking table
Cultivate 10 to 12 hours.100ml LB solid medium is made, suitable temperature is cooled after sterilizing, in superclean bench
Detected bacterium bacterium solution (100 microlitres of bacterium solutions are added in the every 100ml solid medium of standard) is added, shakes up inverted plate, stands solidification.?
Plate back side subregion has been solidified, three to four regions are generally divided into, has marked sample added by good each region.It is preferably in each region
Oxford cup steadily is put in the part of centre, light to press, but guarantees cup cannot be allowed to be inserted into plate.After putting well, each
(sample is 17 obtained in embodiment 3 to the sample of 100 microlitres to 200 microlitres of addition in cup#The metabolism of gingko endogenous fungus
Product, 18#The metabolite product of gingko endogenous fungus).Smoothly plate is put into 37 DEG C of incubators, 9 to 12
Hour carries out result observation.
Note: the disclosure has chosen a variety of gingko endogenous fungus and is tested, and gingko endogenous fungus is numbered, and chooses
Relatively good experimental result is presented in the disclosure, wherein 17#、18#Respectively corresponding CN108102928A, " one plant gingko endogenous
Fungi and its application " in J-1 and J-2,17#Bacterial strain is the raw sickle-like bacteria (Fusarium of gingko endogenous fungus layer
Proliferatum) DZHQ1, deposit number are CGMCC No.14983.
2, experimental result:
2.1 pairs of Escherichia coli inhibitory effects
Fig. 5 is the gingko endogenous fungus 17 in embodiment 3#、18#The methanol solution and methanol of crude extract are to Escherichia coli
The experimental result of inhibiting effect.Wherein 17#Represent gingko endogenous fungus 17#Crude extract methanol solution；18#What is represented is silver
Apricot endogenetic fungus 18#Crude extract methanol solution；What " first " represented is methanol solution.It can be found that 17 from figure#It is gingko endogenous
Fungi, antibacterial circle diameter are 0.9cm.18#The inhibitory effect of endogenetic fungus and methanol solution is very faint, almost without.
Fig. 6 is inhibitory effect of the aqueous solution to Escherichia coli of the gingko endogenous fungus crude extract in embodiment 3.Wherein
17#B represents gingko endogenous fungus 17#Crude extract aqueous solution；18#What B was represented is gingko endogenous fungus 18#Crude extract
Aqueous solution；What " water " represented is aqueous solution.It can be seen from the figure that both without fungistatic effect.
In conclusion gingko endogenous fungus 17#、18#In, 17#Endogenetic fungus is more apparent to the inhibitory effect of Escherichia coli,
Followed by 18#The effect of endogenetic fungus is not it is obvious that effect is almost 0.In addition, it can also be seen that, difference is molten from the experiment
The inhibitory effect to Escherichia coli for the metabolite that agent is extracted is different.17#Suppression of the methanol solution of crude extract to Escherichia coli
Effect processed is more obvious.
2.2 pairs of staphylococcus aureus inhibitory effects
Using 17 in embodiment 3#Sample or 18#Sample carries out Oxford cup bacteriostatic test.As a result as follows:
Fig. 7 is the gingko endogenous fungus crude extract methanol solution in embodiment 3 to staphylococcus aureus inhibitory effect.It is real
It tests in result, 17#With 18#There is certain inhibitory effect, antibacterial circle diameter is respectively 2cm and 2.9cm.
Fig. 8 is the gingko endogenous fungus crude extract aqueous solution in embodiment 3 to staphylococcus aureus inhibitory effect, is passed through
Although observation is not it can be found that some inhibition zones are apparent, it can be seen that 17#Endogenous fungus metabolite has certain
Fungistatic effect, 18#Endogenous fungus metabolite fungistatic effect be it is faint, almost without.
The results show that 17#With 18#Gingko endogenous fungus has certain inhibitory effect to staphylococcus aureus.From the experiment
It can also be seen that the inhibitory effect to staphylococcus aureus for the metabolite that different solvents extract is different.17#The first of crude extract
Alcoholic solution and aqueous solution have obvious inhibitory effect to staphylococcus aureus.
Above-described embodiment is the preferable embodiment of the disclosure, but embodiment of the present disclosure is not by above-described embodiment
It limits, made changes, modifications, substitutions, combinations, simplifications under other any spiritual essence and principles without departing from the disclosure,
It should be equivalent substitute mode, be included within the protection scope of the disclosure.
Claims (10)
1. the metabolite product of one plant of gingko endogenous fungus, it is characterized in that: fungi used is the raw sickle of gingko endogenous fungus layer
Knife bacterium (Fusarium proliferatum) DZHQ1, deposit number is CGMCC No.14983, on November 28th, 2017
It is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, fermented culture and ethyl acetate obtain after extracting
It to the ethyl acetate extract of fermentation culture, is then dissolved using methanol or water, gingko endogenous fungus is obtained after dissolution
Metabolite product.
2. metabolite product as described in claim 1, it is characterized in that: including glycine betaine, Scopoletin, harmine
Alkali, Rosmarinic acid, oxipurinol, resveratrol, naringenin, catechin, texifolin and xanthohumol；
Further, glycine betaine, Scopoletin, yageine, Rosmarinic acid, oxipurinol, resveratrol, naringenin,
Catechin, texifolin and xanthohumol are peak area in HPLC figure 105The substance that above peak represents.
3. the preparation method of the metabolite product of one plant of gingko endogenous fungus, characterized in that this method comprises:
Take cultured gingko endogenous fungus (Fusarium proliferatum) DZHQ1, preservation in PDA solid medium
Number is CGMCC No.14983, by inoculated by hypha block in PDA liquid medium, 20~28 DEG C, 100~150r/min shaking table
Cultivate 5~7d；Then ethyl acetate is added by the volume ratio of 1:1~2:1, continues to cultivate 4~8d in shaking table；Fermentation liquid passes through
Mycelium is removed after filter, then is layered, and the organic phase containing gingko endogenous fungus secondary metabolite is obtained；Recycle acetic acid second
Ester obtains the concentrate containing gingko endogenous fungus secondary metabolite；Finally, concentrate is dry, as fermentation culture
Ethyl acetate extract, then dissolved using methanol or water, obtain methanol after dissolution or water is dissolved gingko endogenous
The metabolite product of fungi.
4. using the metabolite product for the gingko endogenous fungus that method as claimed in claim 3 is prepared.
5. the metabolite product of gingko endogenous fungus described in claims 1 or 2 or 4 is preparing answering in antimicrobial product
With.
6. application as claimed in claim 5, it is characterized in that: the product is drug.
7. application as claimed in claim 5, it is characterized in that: the metabolite of the gingko endogenous fungus can resist Escherichia coli
And/or staphylococcus aureus.
8. a kind of bactericidal composition, it is characterized in that: being with the metabolite of gingko endogenous fungus described in claims 1 or 2 or 4
Active constituent.
9. composition as claimed in claim 8, it is characterized in that: composition further includes pharmaceutically acceptable carrier.
10. a kind of antibiotic method, it is characterized in that: this method includes being produced using the metabolism of gingko endogenous fungus described in claim 4
Bactericidal composition described in produce product or claim 8 inhibits the step of bacterium, wherein the bacterium is Escherichia coli and/or gold
Staphylococcus aureus.
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| CN201811004688.5A CN109082445B (en) | 2018-08-30 | 2018-08-30 | Metabolite product of ginkgo endophytic fungi and application of metabolite product in antibiosis |
| US16/469,830 US11332707B2 (en) | 2017-12-21 | 2018-11-02 | Endophytic fungus from gingko, metabolite product and use thereof |
| PCT/CN2018/113709 WO2019119992A1 (en) | 2017-12-21 | 2018-11-02 | Endophytic fungus of ginkgo biloba and metabolite product and application thereof |
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| WO2022188162A1 (en) * | 2021-03-12 | 2022-09-15 | 德州学院 | Endophytic fungus in ginkgo biloba and anti-bacterial and antitumor application thereof |
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