CN102174051A - Strong algae activity inhibiting compound as well as preparation method and application thereof - Google Patents
Strong algae activity inhibiting compound as well as preparation method and application thereof Download PDFInfo
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Abstract
The invention provides a strong algae activity inhibiting compound as well as a preparation method and application thereof and relates to an algae killing compound. The strong algae activity inhibiting compound is called as nigericin and has the molecular formula of C40H68O11 and the relative molecular mass of 742; a streptomyces malaysiensis O4-6 is obtained by screening through a conventional method, is preserved in the China Center for Type Culture Collection (CCTCC) at February 28th, 2011 and has the preservation number of CCTCC No. M2011051. The preparation method comprises the following steps of: carrying out fermentation culture to obtain fermentation liquid containing the strong algae activity inhibiting compound; centrifuging the fermentation liquid; collecting supernatant fluid; then separating and purifying the supernatant fluid to obtain the strong algae activity inhibiting compound. The strong algae activity inhibiting compound can efficiently and exclusively kill algae cells, has proficiency for developing an algae inhibiting agent and has wide application in aspects of algae biodegradation and red tide management.
Description
Technical field
The present invention relates to algae compound extremely, particularly a kind of potent algistatic activity compound and preparation method thereof and application.
Background technology
In recent years, along with being extensive use of of molecular ecology method, the huge progress that the actinomycetic Study on Diversity in marine actinomycete Study on Diversity, especially deep-sea obtains.Increasing marine actinomycete be found (1, Prof.Alan Claude, W., Diversity and biogeography of marine actinobacteria[J] .2006.9 (3): 279-286.), comprise new 16SrDNA sequence cluster (2, Liesack, W.and E.Stackebrandt, Occurrence of novel groups of the domain Bacteria as revealed by analysis of genetic material isola
Because the singularity of ocean environment, marine microorganism have unique metabolic way, the metabolic chemistry structure of generation has great complicacy and diversity.Over nearly 20 years, the report that relevant marine microorganism produces the secondary metabolite of new biologically active increases gradually, finds the probability of new biological activity material even surpasses the Lu Sheng actinomycetes from marine actinomycete.The part active substance that obtains from marine actinomycete has griseorhodin A, salt spore amine A, and salt spore amine B, smoked clothing cyanines are blue plain, salt amine, tetraodotoxin, ocean mycin A, B, four and mycin D1, chromomycin A3, enterococcin, Quinomycin A, antimycin A, crust bifilomycin, Antibiotic U-5956, lipomycin, the lagosin, Deng (6, Xu Lihua etc., actinomycetes systematics---principle, method and put into practice [M]. Beijing: Science Press, 2007.).These results of study show that ocean environment is the precious resources storehouse of finding and screening new actinomycetes species and meta-bolites, and wherein containing in a large number, the actinomycetes resources await development and utilize.
Since 20th century, be accompanied by the coastland explosive population growth, industrial or agricultural develop serious organic contamination of inner bay, river mouth and the stretch of coastal water and eutrophication rapidly.Harmful algae wawter bloom (Harmful algae blooms, HABs) generation scale and the frequency in China is rapid ascendant trend, to China coastal caused serious ecology, resource, environmental problem and great financial loss (7, Qin, S., et al., Two New Metabolites, Epoxydine A and B, from Phoma sp.Helvetica Chimica Acta[J], 2010.93 (1): 169-174).Research red-tide control method is formulated the environmental practice that relevant Preventing Countermeasures has become the task of top priority of many countries and regions.Because physics and chemical process are administered red tide and are existed and be difficult to that big area is used and easily cause weak point such as secondary pollution, utilize biological Ecology Action each other administer red tide become the current research focus (8, Skerratt, J.H., et al., Algicidal bacteria associated with blooms of a toxic dinoflagellate in a temperateAustralian estuary.Marine Ecology-Progress Series[J], 2002.244:1-15.).
Up to now, the algal control microorganism that screens of great majority is to have come the algal control effect by secreting the special material with algistatic activity.The algistatic activity material of having reported comprises: other still not qualitative algal control compounds such as protein (containing extracellular enzyme), polypeptide, amino acid, microbiotic, nitrogenous compound (9, Yoshikawa, K., et al., beta-cyanoalanine production bymarine bacteria on cyanide-free medium and its specific inhibitory activity toward cyanobacteria.Applied and Environmental Microbiology[J], 2000.66 (2): 718-722).Become the new approaches that the exploitation algae-inhibiting agent is used for red tide improvement by screening efficient, single-minded algistatic activity material.
Summary of the invention
First purpose of the present invention provides a kind of potent algistatic activity compound.
Second purpose of the present invention provides a kind of bacterial strain of producing potent algistatic activity compound.
The 3rd purpose of the present invention provides a kind of preparation method of potent algistatic activity compound.
The 4th purpose of the present invention provides the application of a kind of potent algistatic activity compound in the preparation algae-inhibiting agent.
Described strong algistatic activity compound the name be called polyetherin A (Nigericin), molecular formula is C
40H
68O
11, relative molecular mass is 742, structural formula is:
The production bacterial strain of described potent algistatic activity compound is Streptomyces malaysiensis O4-6, this bacterial strain has been preserved in Chinese typical culture collection center on February 28th, 2011, preservation center deposit number is: CCTCC NO:M2011051, address: China. Wuhan. and Wuhan University.
The preparation method of described potent algistatic activity compound may further comprise the steps:
1) Streptomyces malaysiensis O4-6 is inoculated in the dull and stereotyped line separation of going up, under 28～37 ℃ of temperature, cultivates 5～7d; Picking list colony inoculation is in the Gause I liquid nutrient medium, and in 28～37 ℃, 180～250rpm promptly gets fermented liquid after cultivating 5～7d; The gained fermented liquid is carried out centrifugal, collect supernatant liquor;
2) with ethyl acetate extraction step 1) gained supernatant liquor, concentrating under reduced pressure, drying gets acetic acid ethyl ester extract;
3) acetic acid ethyl ester extract is dissolved in ethyl acetate, joins again and carry out wash-out in the silicagel column, collect elutriant and carry out thin layer chromatography analysis,, merge the elutriant of collecting according to launching collection of illustrative plates;
4) with step 3) gained elutriant in 30～45 ℃ of following evaporated in vacuo, product carries out the mensuration of algistatic activity, but obtains the active ingredient of algal control;
5) but the active ingredient of gained algal control is carried out the high performance liquid chromatography wash-out, collect elutriant, obtain potent algistatic activity compound.
In step 1), described centrifugal condition can be: speed 10000～12000g, time 10～20min.
In step 3), the specification of described silicagel column can be 170mm * 30mm, 200～300 orders; The eluent of described wash-out can be the mixture of normal hexane and ethyl acetate etc.; The program of described wash-out is: with normal hexane and 1: 2 mixture wash-out 20min of ethyl acetate volume ratio, use 1: 1 mixture wash-out 30min of normal hexane and ethyl acetate volume ratio again, use normal hexane wash-out 30min at last; The flow velocity of wash-out can be 1mL/min; The developping agent of described thin layer chromatography analysis can be ethyl acetate etc., and the developer of described thin layer chromatography analysis is chloroformic solution that contains 0.5% iodine etc.
In step 5), the program of described wash-out can be:
0～7.5min, 85% methanol (v/v);
7.5～10.5min 85% methanol～100% methyl alcohol;
10.5～22.5min 100% methyl alcohol.
Collect 10.5～12.5min elutriant and be dissolved in DMSO checking algistatic activity in 30 ℃ of following evaporated in vacuo of Rotary Evaporators.
The potent algistatic activity compound of gained is through algistatic activity component thin-layer chromatography (Thin Layer Chromatography, TLC) detect, on silica gel thin-layer plate, launch to be a spot as developping agent with ethyl acetate, methylene dichloride and chloroform, be defined as pure compound, and warp
1H-NMR detects, and determines the structural formula of potent algistatic activity compound, and described potent algistatic activity compound can effectively be killed the phaeocystis globosa cell, and it has huge application potential in the preparation of algae-inhibiting agent.
The present invention uses the ordinary method screening to obtain a strain Streptomyces malaysiensis O4-6, pass through fermentation culture, acquisition contains the fermented liquid of strong algistatic activity compound, described fermented liquid is centrifugal, collect supernatant liquor, then described supernatant liquor is carried out separation and purification, obtain to have the compound of strong algistatic activity.The compound of described strong algistatic activity can be killed frustule efficient, single-mindedly, has the potential that is developed to algae-inhibiting agent, has widely at aspects such as biological degradation algae and improvement red tides and uses.
Description of drawings
Fig. 1 is the high performance liquid phase collection of illustrative plates of potent algistatic activity compound.In Fig. 1, X-coordinate is time (min), and ordinate zou is absorbancy (mAU); Respectively compose the peak from left to right and be respectively 2.948,10.491,10.910,12.042.
Embodiment
Following examples are to further specify of the present invention, but the invention is not restricted to following embodiment.
The separation screening of embodiment 1 marine streptomyces Streptomyces malaysiensis O4-6
One, the separation screening concrete steps of Streptomyces malaysiensis O4-6 are:
1) (particular location is Fujian sky mangrove forest wetland: 117 ° 24 '-117 ° 30 ' E, 23 ° 53 '-23 ° 56 ' N) settling, placed dry 1 month in room temperature, get the dry soil sample of 10g, be dissolved in autoclaved 90mL Gause I substratum, place 150rpm shaking table concussion 20min, make the soil sample homodisperse;
2), coat Gause I (20g Zulkovsky starch, 1g NaNO with 10 times of dilutions of step 1) gained sample
3, 0.5gK
2HPO
4, 0.5g MgSO
47H
2O, 0.01g FeSO
47H
2O, 75 μ g K
2Cr
2O
7, 10g agar, 1L seawater) and solid plate, place under 28 ℃ of temperature and cultivate 5d;
3) the dissimilar single bacterium colonies of picking line the Gause I solid plate, place 28 ℃ to cultivate down 5d, verify whether pure culture, repeat this step up to obtaining pure culture;
4) the isolated pure growth list bacterium colony of inoculation places 28 ℃ of shaking tables in 4mL Gause I liquid nutrient medium, and 5d is cultivated in the 180rpm concussion, gets culture centrifugal 10min under the centrifugal force of 10000g, removes bacterial sediment, and supernatant is saved to the 4.5mL centrifuge tube;
5) the 1mL supernatant is joined in the 20mL exponential phase phaeocystis globosa nutrient solution, in 20 ± 1 ° ℃, 12h illumination, 12h dark, 50 μ mol photons m
-2s
-1Cultivate 2d under the intensity of illumination condition, the Gause I substratum is for contrast adds in the algae liquid, be provided with respectively 3 parallel; Observe the whether sedimentation of phaeocystis globosa cell, if sedimentation then represent in the 7d culture supernatant of thalline and contain the algistatic activity material, thus filter out the algistatic activity bacterial strain.
Two, the separation screening concrete steps of Streptomyces malaysiensis O4-6 can be:
1) Fujian sky mangrove forest wetland (placed dry 1 month in room temperature by 117 ° 24 '-117 ° 30 ' E, 23 ° 53 '-23 ° 56 ' N) settling, get the dry soil sample of 10g, be dissolved in autoclaved 90m L Gause I substratum, place 200rpm shaking table concussion 30min, make the soil sample homodisperse;
2), coat Gause I (20g Zulkovsky starch, 1g NaNO with 10 times of dilutions of step 1) gained sample
3, 0.5gK
2HPO
4, 0.5g MgSO
47H
2O, 0.01g FeSO
47H
2O, 75 μ g K
2Cr
2O
7, 10g agar, 1L seawater) and solid plate, place 37 ℃ to cultivate 7d down;
3) the dissimilar single bacterium colonies of picking line the Gause I solid plate, place 37 ℃ to cultivate down 7d, verify whether pure culture, repeat this step up to obtaining pure culture;
4) the isolated pure growth list bacterium colony of inoculation places 37 ℃ of shaking tables in 4mL Gause I liquid nutrient medium, and 7d is cultivated in the 250rpm concussion, gets culture centrifugal 20min under the centrifugal force of 12000g of 7d, removes bacterial sediment, and supernatant is saved to the 4.5mL centrifuge tube;
5) the 1mL supernatant is joined in the 20mL exponential phase phaeocystis globosa nutrient solution, in 20 ± 1 ℃, 12h illumination, 12h dark, 50 μ mol photons m
-2s
-1Cultivate 2d under the intensity of illumination condition, the Gause I substratum is for contrast adds in the algae liquid, be provided with respectively 3 parallel; Observe the whether sedimentation of phaeocystis globosa cell, if sedimentation then represent in the supernatant of thalline 7d culture and contain the algistatic activity material, thus filter out the algistatic activity bacterial strain.
The method of calculation of embodiment 2 algal control rates
1) phaeocystis globosa is at 20 ± 1 ℃, 12h illumination, 12h dark, 50 μ mol photons m
-2s
-1Under the intensity of illumination condition, be cultured to exponential phase in triangular flask, divide then to install in the 24 porocyte culture plates, every hole packing 2mL frustule suspension adapts to growth 1d:
2) 100 μ L culture to be measured adds 24 orifice plates, cultivates 2d;
3) get the phaeocystis globosa nutrient solution, 200 μ L samples with the fluorescence intensity at 680nm place under the microplate reader detection 460nm excitation, calculate algal control rate according to following formula in 24 orifice plates, observe the frustule metamorphosis simultaneously:
Algal control rate=(F
C-F
T)/F
C* 100%
In the formula, F
CExpression control group fluorescence intensity, F
TExpression experimental group fluorescence intensity.
Embodiment 3 algistatic activity physical properties are identified
1) marine streptomyces Streptomyces malaysiensis O4-6 is inoculated in 28～37 ℃ of shaking tables of Gause I substratum, and 5～7d is cultivated in 180～250rpm concussion.Fermented liquid is centrifugal 10～20min under the centrifugal force of 10000～12000g, obtains streptomycete Streptomyces malaysiensis O4-6 nutrient solution supernatant; With ethyl acetate extraction gained supernatant liquor, concentrating under reduced pressure, drying, get acetic acid ethyl ester extract.
2) Temperature Treatment experimental design: the nutrient solution supernatant that contains the algistatic activity material is carried out 40 ℃, 60 ℃, 80 ℃, 100 ℃ and 120 ℃ of high pressure 30min thermal treatment, 3 parallel laboratory test groups are set; The control experiment group is: the nutrient solution supernatant is untreated; Get 100 μ L nutrient solution supernatants and add in the 2mL algae culturing liquid, behind the cultivation 2d, sampling is pressed embodiment 2 and is calculated the algal control efficient.Experimental result (referring to table 1) shows: after this nutrient solution supernatant carried out thermograde thermal treatment, its algal control efficient to phaeocystis globosa did not have very big variation, illustrated that the algistatic activity material can not tolerate pyroprocessing more than 100 ℃.
Table 1 Temperature Treatment is to the influence of algistatic activity material algal control efficient
3) pH handles experimental design: measure nutrient solution supernatant pH value, pH transfers to 1,3,5,7,9 and 11 respectively with the nutrient solution supernatant, again its pH is recalled to processing behind the 2h, and 3 parallel laboratory test groups are set; Get 100 μ L bacteria-free filtrates and add in the 2mL algae culturing liquid, behind the cultivation 24h, sampling is pressed embodiment 2 and is calculated the algal control efficient.Experimental result (referring to table 2) shows: after this nutrient solution supernatant carried out acidifying and alkalinisation treatment, its algal control efficient to phaeocystis globosa did not have very big variation, illustrated that the algistatic activity material is all more stable under acidic conditions and alkaline condition.
Table 2pH handles the influence to algistatic activity material algal control efficient
4) dialysis treatment experimental design: utilize the molecular retention amount to be about the 1Kd dialysis tubing, the nutrient solution supernatant is carried out dialysis treatment, behind PBS damping fluid dialysis 3h, change fresh PBS damping fluid over to and continue dialysis 3h, 3 parallel laboratory test groups are set; Control group is the nutrient solution supernatant of not dialysing; Get 100 μ L nutrient solution supernatants and add in the 2mL algae culturing liquid, behind the cultivation 24h, sampling is pressed embodiment 2 and is calculated the algal control efficient.Experimental result (referring to table 3) shows: after this nutrient solution supernatant is about the dialysis tubing dialysis treatment of 1Kd through the molecular retention amount, its algal control efficient to phaeocystis globosa obviously reduces, its algal control efficient is 21.4% only on average, illustrates that the algistatic activity molecular weight of material is less than 1Kd.
Table 3 dialysis treatment is to the influence of algistatic activity material algal control efficient
5) different organic solvents extraction experiments design: select propyl carbinol, methylene dichloride, ethyl acetate as extraction agent, with 1: 1 volume ratio extraction 500mL nutrient solution supernatant, repeat 3 extractions respectively.500mL nutrient solution supernatant is selected the dissolving of 500mL acetonitrile in 30 ℃ of following evaporated in vacuo of Rotary Evaporators.Various extractive with organic solvent obtain the extraction crude extract in 30 ℃ of following evaporated in vacuo of Rotary Evaporators.5mL DMSO dissolves crude extract, gets 100 μ L DMSO solutes and adds in the 2mL algae culturing liquid, and behind the cultivation 24h, sampling is pressed embodiment 2 and calculated the algal control efficient, and 3 parallel laboratory test groups are set, and adds DMSO to the algae culturing liquid control group.Experimental result (table 4) shows: behind this nutrient solution supernatant process opposed polarity organic solvent extraction, algal control efficient shows certain difference, acetic acid ethyl ester extract algal control rate is higher, and average out to 87.9% illustrates that algistatic activity material polarity is more weak to be come out by ethyl acetate extraction easily.
Table 4 different organic solvents extract algal control efficient
Embodiment 4 algistatic activity component silica gel column chromatographies
Streptomycete Streptomyces malaysiensis O4-6 is inoculated in 28～37 ℃ of shaking tables of Gause I substratum, and 5～7d is cultivated in 180～250rpm concussion.Fermented liquid is centrifugal 10～20min under the centrifugal force of 10000～12000g, obtains streptomycete Streptomyces malaysiensis O4-6 nutrient solution supernatant.With ethyl acetate extraction gained supernatant liquor, concentrating under reduced pressure, drying, get acetic acid ethyl ester extract.
The 100mg acetic acid ethyl ester extract is dissolved in the 1mL ethyl acetate, is splined on silicagel column (170 * 30mm, 200-300 order), eluent: the mixture of normal hexane and ethyl acetate, elution program: volume ratio 1: 2,20min; 1: 1,30min; 1: 0,30min; Elution flow rate: 1mL/min; Collect elutriant 2mL/ pipe with collection tube.
(Thin Layer Chromatography TLC) analyzes embodiment 5 algistatic activity component thin-layer chromatographys
1) as embodiment 4, elutriant utilizes TLC to analyze, and developping agent is an ethyl acetate, developer: the chloroformic solution of 0.5% iodine merges elutriant in the collection tube according to launching collection of illustrative plates.
2) merge elutriant in 30 ℃ of following evaporated in vacuo of Rotary Evaporators, be dissolved in DMSO, press the method validation algistatic activity among embodiment 2, the embodiment 3, obtain potent algistatic activity component.
(High-performance liquid chromatography HPLC) analyzes embodiment 6 algistatic activity component high performance liquid chromatography
The potent algistatic activity component of embodiment 5 gained utilizes HPLC to analyze elution program:
0～7.5min, 85% methanol;
7.5min～10.5min 85% methanol～100% methyl alcohol;
10.5～22.5min 100% methyl alcohol.
Experimental result as shown in Figure 1, the elutriant of collecting 10.5～12.5min is dissolved in DMSO in 30～45 ℃ of following evaporated in vacuo of Rotary Evaporators, press embodiment 2, embodiment 3 verifies algistatic activities.
The structure of embodiment 7 potent algistatic activity compounds is identified:
Embodiment 6 algistatic activities detect, and detect through TLC, on silica gel thin-layer plate, launch to be a spot with ethyl acetate, methylene dichloride and chloroform, and warp
1H-NMR detects, and is defined as pure compound, and structural formula is as follows:
The ESI-MS spectrum of described pure compound is at m/z 747.47[M+Na]
+The place shows the adduct ion peak of molecule, and in conjunction with 13C-NMR (DEPT) spectrum and 1H-NMR spectrum, releasing its molecular formula is C
40H
68O
11, relative molecular mass is 742.The 1H-NMR spectrum shows a large amount of methyl, methylene radical and 1 methoxyl group signal.13C-NMR spectrum show 40 C, by with the collection of illustrative plates of DEPT relatively, can obviously obtain: 10 CH
3(δ
CBe respectively 59.53,29.00,22.76,16.98,16.41,16.12,14.38,13.39,13.07,11.48), 10 CH
2(δ
CBe respectively 66.88,41.65,37.08,35.77,32.30,31.99,29.47,26.27,25.86,23.51), 15 CH
1(δ
CBe respectively 85.17,81.39,79.40,76.75,76.43,73.20,68.37,60.38,45.79,39.58,36.69,36.40,35.07,31.80,27.65), 5 quaternary carbons (δ C is respectively 183.83,107.49,97.15,84.78,82.31).From the two-dimensional map of HMBC, HSQC, H-H COSY and NOSEY, can determine the relevant ownership of each C and H.Through the Scifinder database retrieval, with corresponding document comparison, comprehensive then its physico-chemical property is determined consistent with polyetherin A again.
In sum, to extract the compound with algistatic activity that purifying obtains be polyetherin A in the present invention.
Claims (9)
1. a potent algistatic activity compound is characterized in that its name is called polyetherin A, and molecular formula is C
40H
68O
11, relative molecular mass is 742, structural formula is:
2. the production bacterial strain of a kind of potent algistatic activity compound as claimed in claim 1, it is characterized in that it is Streptomyces malaysiensis O4-6, this bacterial strain is preserved in Chinese typical culture collection center on February 28th, 2011, and preservation center deposit number is: CCTCC NO:M2011051.
3. the preparation method of a kind of potent algistatic activity compound as claimed in claim 1 is characterized in that may further comprise the steps:
1) Streptomyces malaysiensis O4-6 is inoculated in the dull and stereotyped line separation of going up, under 28～37 ℃ of temperature, cultivates 5～7d; Picking list colony inoculation is in the Gause I liquid nutrient medium, and in 28～37 ℃, 180～250rpm promptly gets fermented liquid after cultivating 5～7d; The gained fermented liquid is carried out centrifugal, collect supernatant liquor;
2) with ethyl acetate extraction step 1) gained supernatant liquor, concentrating under reduced pressure, drying gets acetic acid ethyl ester extract;
3) acetic acid ethyl ester extract is dissolved in ethyl acetate, joins again and carry out wash-out in the silicagel column, collect elutriant and carry out thin layer chromatography analysis,, merge the elutriant of collecting according to launching collection of illustrative plates;
4) with step 3) gained elutriant in 30～45 ℃ of following evaporated in vacuo, product carries out the mensuration of algistatic activity, but obtains the active ingredient of algal control;
5) the gained active ingredient is carried out the high performance liquid chromatography wash-out, collect elutriant, obtain potent algistatic activity compound.
4. the preparation method of a kind of potent algistatic activity compound as claimed in claim 3 is characterized in that in step 1), and described centrifugal condition is: speed 10000～12000g, time 10～20min.
5. the preparation method of a kind of potent algistatic activity compound as claimed in claim 3 is characterized in that in step 3), and the specification of described silicagel column is 170mm * 30mm, 200～300 orders.
6. the preparation method of a kind of potent algistatic activity compound as claimed in claim 3, it is characterized in that in step 3), the eluent of described wash-out is the mixture of normal hexane and ethyl acetate, the program of described wash-out is: with normal hexane and 1: 2 mixture wash-out 20min of ethyl acetate volume ratio, use 1: 1 mixture wash-out 30min of normal hexane and ethyl acetate volume ratio again, use normal hexane wash-out 30min at last, the flow velocity of wash-out is: 1mL/min.
7. the preparation method of a kind of potent algistatic activity compound as claimed in claim 3 is characterized in that in step 3), and the developping agent of described thin-layer chromatography is an ethyl acetate, and the developer of described thin-layer chromatography is the chloroformic solution that contains 0.5% iodine.
8. the preparation method of a kind of potent algistatic activity compound as claimed in claim 3 is characterized in that in step 5), the program of described wash-out:
0～7.5min, 85% methanol;
7.5～10.5min 85% methanol～100% methyl alcohol;
10.5～22.5min 100% methyl alcohol.
9. the application of a kind of potent algistatic activity compound as claimed in claim 1 in the preparation algae-inhibiting agent.
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| CN105368892B (en) * | 2015-11-05 | 2018-12-18 | 桂林电子科技大学 | It is a kind of for improving the fermentation medium and fermentation process of nonactin yield |
| CN106117238B (en) * | 2016-06-24 | 2018-08-21 | 福建省微生物研究所 | The purification process of nigericin |
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| CN103145725B (en) * | 2011-12-07 | 2016-02-24 | 上海来益生物药物研究开发中心有限责任公司 | A kind of compound and its preparation method and application |
| CN102911972A (en) * | 2012-11-12 | 2013-02-06 | 厦门大学 | Preparation method of algistatic active substance di-isobutoxyphenyl amine |
| CN102911972B (en) * | 2012-11-12 | 2015-01-21 | 厦门大学 | Preparation method of algistatic active substance di-isobutoxyphenyl amine |
| CN105314739A (en) * | 2014-08-19 | 2016-02-10 | 湖北工业大学 | Microbial algicidal agent synchronous nitrogen removing and organic matter removing method |
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| WO2020121324A3 (en) * | 2018-12-10 | 2020-07-23 | Council Of Scientific & Industrial Research | A process for production of nigericin from streptomyces sp. mcc-0151 |
| CN113396214A (en) * | 2018-12-10 | 2021-09-14 | 科学与工业研究委员会 | Method for producing nigericin from streptomyces sp.mcc-0151 |
| US20220025418A1 (en) * | 2018-12-10 | 2022-01-27 | Council Of Scientific & Industrial Research | A process for production of nigericin from streptomyces sp. mcc-0151 |
| US12018305B2 (en) * | 2018-12-10 | 2024-06-25 | Council Of Scientific & Industrial Research | Process for production of nigericin from Streptomyces sp. MCC-0151 |
| CN113396214B (en) * | 2018-12-10 | 2024-07-02 | 科学与工业研究委员会 | Method for producing nigericin from streptomyces sp.mcc 0151 |
| CN113527325A (en) * | 2021-07-19 | 2021-10-22 | 中国科学院南海海洋研究所 | Nigericin derivative, preparation method thereof and application of nigericin derivative in preparation of antitumor drugs |
| CN113527325B (en) * | 2021-07-19 | 2023-11-17 | 中国科学院南海海洋研究所 | Nigericin derivative, preparation method thereof and application thereof in preparation of antitumor drugs |
| CN116496923A (en) * | 2022-11-30 | 2023-07-28 | 南京灿辰微生物科技有限公司 | Streptomyces griseoviridis 0728-09 and application thereof |
| CN116496923B (en) * | 2022-11-30 | 2024-07-19 | 南京灿辰微生物科技有限公司 | Streptomyces griseoviridis 0728-09 and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103540547A (en) | 2014-01-29 |
| CN103497980B (en) | 2015-08-05 |
| CN103540547B (en) | 2016-03-09 |
| CN103497980A (en) | 2014-01-08 |
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