CN103695342B - One strain has genus bacillus and the application thereof of molten algae activity - Google Patents
One strain has genus bacillus and the application thereof of molten algae activity Download PDFInfo
- Publication number
- CN103695342B CN103695342B CN201310683393.6A CN201310683393A CN103695342B CN 103695342 B CN103695342 B CN 103695342B CN 201310683393 A CN201310683393 A CN 201310683393A CN 103695342 B CN103695342 B CN 103695342B
- Authority
- CN
- China
- Prior art keywords
- lzh
- genus bacillus
- algae
- blue
- bacillus
- 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
Landscapes
- Agricultural Chemicals And Associated Chemicals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a strain there is the genus bacillus of molten algae activity and controlling the application in blue-green alga bloom.Be separated from the water body of Taihu Lake and obtain genus bacillus (Bacillus sp.) Lzh-5 that a strain has remarkable molten algae activity, preserving number is CGMCC No.8282, and separation and purification from its meta-bolites also identifies its effective molten algae composition hexahydropyrrolo also [1, 2-a] pyrazine-1, 4-diketone and 3-sec.-propyl-hexahydropyrrolo also [1, 2-a] pyrazine-1, 4-diketone, wherein hexahydropyrrolo also [1, 2-a] pyrazine-1, 4-diketone and 3-sec.-propyl-hexahydropyrrolo also [1, 2-a] pyrazine-1, the Lethal Dose 50 LD50 of 4-diketone to microcystic aeruginosa 9110 is respectively 5.7 μ g/mL and 19.4 μ g/mL.Can be used for research and development and the production of new bio algicide, be finally applied to the control of lake blue algae wawter bloom.
Description
Technical field
The present invention relates to field of environment microorganism, particularly genus bacillus (Bacillussp.) Lzh-5 of a strain secretion algicidal substances and the application in blue-green alga bloom controls thereof.
Background technology
The control techniques of current blue-green alga bloom can be summarized as physical method, chemical process and biological method.Physical method as machinery except algae, flocculation are sent out except algae and electromagnetic field can as the subcontrol measures of algal bloom except algae etc., but shortcoming is that cost is very high, equipment is complicated, cures the symptoms, not the disease, and processing power is limited; Chemical process such as the chemical algicides such as weedicide directly can kill algae, but the specificity of these chemical substances is poor, large to aquatic toxicity, and easily cause objectionable impurities to remain and secondary pollution.
The defects such as the high cost that physically based deformation method and chemical process control to exist in wawter bloom, highly difficult, high pollution, biological method, because of its Huan Bao ﹑ economic dispatch advantage, receives increasing concern.Algae-lysing bacterium (algae-lysing bacteria) for blue-green algae is the important component part of freshwater environment system, to the biomass reducing blue-green algae, maintains the eubiosis and has vital role.
Therefore, those skilled in the art is devoted to the efficient olution-type adhesive screening efficient algae-lysing bacterium or separation and concentration algae-lysing bacterium metabolism generation, to develop microorganism algicide, in order to the control blue-green alga bloom problem of safety and efficiently.
Summary of the invention
In view of physical method in prior art with chemical process processing power is limited, the defect that easily causes secondary pollution, this technical problem to be solved finds the efficient olution-type adhesive of efficient algae-lysing bacterium and separation and concentration algae-lysing bacterium metabolism generation, in order to the control blue-green alga bloom problem of safety and efficiently.
For achieving the above object, the invention provides genus bacillus and the application of meta-bolites in blue-green alga bloom controls thereof that a strain has molten algae activity.
The invention discloses genus bacillus (Bacillus sp.) Lzh-5 that a strain has molten algae activity, Classification And Nomenclature is bacillus sp., be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, depositary institution's title is abbreviated as CGMCC, preserving number CGMCC No.8282, preservation date is on September 27th, 2013.Preservation organization address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica, postcode: 100101, phone: 86-10-64807355.
Further, the invention provides above-mentioned genus bacillus Lzh-5 and control the application in blue-green alga bloom.
Further, the tunning that the invention provides above-mentioned genus bacillus Lzh-5 is controlling the application in blue-green alga bloom.
Further, to the invention discloses in the tunning of above-mentioned genus bacillus Lzh-5 the effective constituent that two kinds have molten algae activity, be respectively the compound with structure shown in structural formula (I) and (II), the invention provides the application of compound in control blue-green alga bloom that one has structure shown in structural formula (I) and (II).
Further, the invention provides a kind of molten phycomycete agent, it is characterized in that, in described molten phycomycete agent, comprise genus bacillus according to claim 1 (Bacillus sp.) Lzh-5.
Further, the invention provides a kind of molten algae medicament, it is characterized in that, comprise the tunning of genus bacillus according to claim 1 (Bacillus sp.) Lzh-5 in described molten algae medicament, described tunning is fermented liquid, concentrate of fermentation liquid, fermentation broth coarse extract or fermentation broth extract.
Further, the invention provides a kind of method controlling blue-green alga bloom, comprise the steps:
1), ferment genus bacillus Lzh-5 bacterial strain;
2), extractive fermentation liquid;
3), extraction liquid evaporate to dryness obtains crude extract;
4), crude extract water-soluble after for controlling blue-green alga bloom.
Preferably, in step 1), fermentation condition is, genus bacillus Lzh-5 is inoculated in the sterilizing beef-protein medium of pH7.0, and 28 DEG C, 200rpm envrionment conditions bottom fermentation 48h, obtain described genus bacillus Lzh-5 fermented liquid.
Preferably, step 2) in, extraction agent is ethyl acetate, and in extraction system, the volume ratio of ethyl acetate and fermented liquid is 1:1, after mixing, puts into vibrator and to vibrate the extraction liquid of 24h, isolated upper strata ethyl acetate solution and genus bacillus Lzh-5 fermented liquid.Fermentation broth coarse extract is after extraction liquid evaporate to dryness.Crude extract, through this area routine techniques means, as organized the method for chromatography, being purified further and can be obtained fermentation broth extract.
Described genus bacillus Lzh-5 fermented liquid is respectively 4 days of following algae strain molten algae rates: microcystic aeruginosa PCC7806 is 88.5 ± 7.4%, microcystic aeruginosa 9110 is 81 ± 5.2%, the algae BN35 that quivers is 83.6 ± 4.5%, Microcystis viridis FACHB-979 is 78.2 ± 9.3%, chroococcoid FACHB-191 is 86.5 ± 7.9%, and chlamydomonas BS3 is 32.5 ± 11.3%; Within 6 days, molten algae rate is respectively: microcystic aeruginosa PCC7806 is 93.7 ± 3.4%, microcystic aeruginosa 9110 is 91.2 ± 6.3%, and the algae BN35 that quivers is 92.7 ± 1.5%, and Microcystis viridis FACHB-979 is 87.1 ± 7.5%, chroococcoid FACHB-191 is 91.4 ± 4.2%, and chlamydomonas BS3 is 23.6 ± 2.7%.
Described genus bacillus Lzh-5 fermented liquid, utilize the meta-bolites of HPLC technology purifying genus bacillus Lzh-5, concrete preparation method is as follows:
Use 0.22 μm of aperture membrane filtration by described extraction liquid evaporate to dryness and after being dissolved in water, filtrate is by the SupersilTM C18-EP semipreparative column of DIKMA company, and water and methyl alcohol are that the HPLC of moving phase carries out preliminary purification, obtain molten algae composition;
By described molten algae composition by DIKMA SupersilTM C18-EP analytical column, water and methyl alcohol are that the HPLC of moving phase is further purified, and obtain two kinds of effective molten algae composition S-5A and S-5B.
In the meta-bolites of described genus bacillus Lzh-5, the chemical structure of effective molten algae composition is analyzed by LC-MS ﹑ GC-MG and NMR and is obtained.
When genus bacillus Lzh-5 is for controlling blue-green alga bloom, the molecular ion peak of its effective molten algae composition S-5A is 155.0821m/z, and molecular weight analyte is 154.0742, and molecular formula is C
7h
10n
2o
2, proton nmr spectra result is
1h NMR (400MHz, D2O) δ 4.22 (s, 1H), 4.06 (dd, J=17.3,2.7Hz, 1H), 3.77 (d, J=17.3Hz, 1H), 3.44 (dd, J=8.7,4.8Hz, 2H), 2.41 – 2.12 (m, 1H), 2.09 – 1.71 (m, 3H); The molecular ion peak of effective molten algae composition S-5B is 197.1276m/z, and molecular weight analyte is 196.1212, and molecular formula is C
10h
16n
2o
2, proton nmr spectra result is
1h NMR (400MHz, D
2o) δ 4.20 (m, 1H), 4.07 (s, 1H), 3.47 (dd, J=16.3,8.9Hz, 1H), 2.38 (m, 1H), 2.26 (d, J=4.7Hz, 1H), 1.97 (s, 1H), 1.83 (s, 1H), 0.98 (d, J=7.2Hz, 1H), 0.76 (d, J=6.9Hz, 1H), carbon-13 nmr spectra result is
13c NMR (101MHz, D
2o) δ 172.23 (s), 167.12 (m), 60.34 (s), 58.72 (m), 45.20 (s), 28.89 (s), 28.10 (s), 21.55 (s), 17.95 (s), 15.06 (s).
Further, effective molten algae composition S-5A is the meta-bolites hexahydropyrrolo also [1 of described genus bacillus Lzh-5,2-a] pyrazine-1,4-diketone, effective molten algae composition S-5B is the meta-bolites 3-sec.-propyl-hexahydropyrrolo also [1 of described genus bacillus Lzh-5,2-a] pyrazine-Isosorbide-5-Nitrae-diketone.
Further, hexahydropyrrolo also [1,2-a] pyrazine-Isosorbide-5-Nitrae-diketone and 3-sec.-propyl-hexahydropyrrolo also [1, the 2-a] pyrazine-Lethal Dose 50 LD50 of Isosorbide-5-Nitrae-diketone to microcystic aeruginosa 9110 be respectively 5.7 μ g/mL and 19.4 μ g/mL.
The algicidal effect of genus bacillus Lzh-5 of the present invention has broad spectrum.Its fermented liquid preparation method is simple, and preparation cycle is short.The fermented liquid of genus bacillus Lzh-5 has good algicidal effect to the microcystic aeruginosa 9110 in Taihu Lake and the algae BN35 etc. that quivers, and microcystic aeruginosa is the main blue-green algae in the blue-green alga bloom of Taihu Lake.Genus bacillus Lzh-5 fermented liquid and acetic acid ethyl acetate extract all can be used for the control of blue-green alga bloom or other micro-algae, and algicidal effect is better.Wherein, genus bacillus Lzh-5 fermented liquid reaches 91.2 ± 6.3% to 6 of microcystic aeruginosa 9110 days molten algae rates.The meta-bolites hexahydropyrrolo of genus bacillus Lzh-5 also [1,2-a] pyrazine-1,4-diketone and 3-sec.-propyl-hexahydropyrrolo also [1,2-a] pyrazine-1,4-diketone has good lethal effect to the main blue-green algae microcystic aeruginosa 9110 in the blue-green alga bloom of Taihu Lake, can be used for research and development and the production of novel algicide, be finally applied to the control of lake blue algae wawter bloom.
Below with reference to accompanying drawing to genus bacillus Lzh-5 bacterial strain of the present invention and meta-bolites hexahydropyrrolo thereof also [1,2-a] pyrazine-1,4-diketone and 3-sec.-propyl-hexahydropyrrolo also [1,2-a] pyrazine-1, the effect of 4-diketone in blue-green alga bloom controls is described further, to understand object of the present invention, characteristic sum effect fully.
Accompanying drawing explanation
Fig. 1 is that molten phycomycete Lzh-5 is to the algicidal effect of microcystic aeruginosa 9110;
Fig. 2 is the algicidal effect that the fermented liquid acetic acid ethyl acetate extract of molten phycomycete Lzh-5 joins on microcystic aeruginosa 9110 algae flat board;
Fig. 3 be algicidal substances hexahydropyrrolo also [1,2-a] pyrazine-Isosorbide-5-Nitrae-diketone and 3-sec.-propyl-hexahydropyrrolo also [1,2-a] pyrazine-Isosorbide-5-Nitrae-diketone join the algicidal effect on microcystic aeruginosa 9110 algae flat board respectively;
Fig. 4 be algicidal substances hexahydropyrrolo also [1,2-a] pyrazine-Isosorbide-5-Nitrae-diketone and 3-sec.-propyl-hexahydropyrrolo also [1,2-a] pyrazine-Isosorbide-5-Nitrae-diketone join the algicidal effect to microcystic aeruginosa 9110 in algae liquid respectively.
Embodiment
The screening of embodiment 1 algae-lysing bacterium
The 10mL natural water samples that Lake Taihu waters gathers is joined in the algae liquid of the microcystic aeruginosa 9110 of 90mL logarithmic phase, get yellow algae liquid gradient dilution method two days later and be coated with beef-protein medium agar plate, cultivate 24h for 28 DEG C, get the flat board that colony density is moderate, select different strains according to the difference of colonial morphology.
Inoculated respectively by the bacterial strain screened in 10mL beef-protein medium, 28 DEG C, 200rpm cultivates 24h, is added respectively by cultured for 10mL bacterium liquid in the algae liquid of 90mL logarithmic phase microcystic aeruginosa.In addition the beef-protein medium after 10mL sterilizing to be added equally in 90mL algae liquid in contrast.Calculate its molten algae efficiency after the algae liquid of all experimental group and control group cultivates 48h in illumination box, the strain bacterium that have chosen wherein molten algae efficiency high is studied further, and this bacterial strain code name is Lzh-5.
Algae liquid culture condition: microcystic aeruginosa 9110 BG11 liquid nutrient medium is cultivated, and is positioned in illumination box, 25 DEG C, intensity of illumination 40 μm of ol photons m
-2s
-1, light dark period is than being 12h:12h (illumination: dark).
Molten algae efficiency calculation method: molten algae efficiency (%)=(control group algae cell density-experimental group algae cell density)/control group algae cell density × 100.Wherein the algae cell density blood counting chamber of microcystic aeruginosa measures.
Microcystic aeruginosa 9110 cell concn curve (molten phycomycete Lzh-5 (starting point concentration 1 × 10 over time when Fig. 1 is for adding molten phycomycete Lzh-5
8cells/mL) (■) and blank (◆)).
The qualification of embodiment 2 genus bacillus Lzh-5 bacterial strain
The Lzh-5 the strongest to algicidal effect by methods such as morphologic observation, dyeing, biochemical reactions, 16srRNA gene sequencings identifies, this bacterial strain is for belonging to G+ bacteria, cell is direct rod shape, 0.5 ~ 2.5 μm × 1.2 ~ 10 μm, normal with paired or catenation, tool nose circle or square end.Cell dyeing great majority present Gram-positive when children cultivates age, move with peritrichous.Gemma ellipse, oval, circle, can resist many poor environments.Have heat, pH and the various multifarious physiological property of salt.Chemoheterotrophic bacteria, tool fermentation or respiratory metabolism type.Through 16srRNA gene sequencing and tetraploid rice, learn that in itself and GenBank, certain Bacillus strain has the homology of 99%, therefore be accredited as bacillus, called after genus bacillus Lzh-5.This bacterial classification has been preserved in national Microbiological Culture Collection management committee common micro-organisms center, preserving number CGMCC No.8282, and preservation date is on September 27th, 2013.Preservation organization address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City institute of microbiology of the Chinese Academy of Sciences, postcode: 100101, phone: 86-10-64807355.The accession number of 16srRNA gene in GenBank of this bacterial classification is HQ896845.
The preparation method of embodiment 3 genus bacillus Lzh-5 fermented liquid and acetic acid ethyl acetate extract thereof
Be inoculated in sterilizing beef-protein medium by genus bacillus Lzh-5 according to 1% inoculum size, at 28 DEG C, 200rpm shaking table obtains genus bacillus Lzh-5 containing fermented liquid after cultivating 48h.Ethyl acetate is added in fermented liquid according to the ratio of 1:1, puts into vibrator and to vibrate 24h, isolate upper solution, i.e. acetic acid ethyl acetate extract.By ethyl acetate evaporate to dryness, be dissolved in water, for being further purified meta-bolites after re-using the filtering with microporous membrane in 0.22 μm of aperture.
Embodiment 4 genus bacillus Lzh-5 is to the algicidal effect of different blue-green algae and Eukaryotic Algae
By 6 parts of cultured (culture condition: beef-protein medium, 28 DEG C, 200rpm cultivates 24h) 10mL genus bacillus Lzh-5 bacterium liquid adds (90mL in the algae liquid of 6 strain blue-green algaes and eukaryotic algae (table 1) respectively, all algae liquid all cultivates logarithmic phase), the aseptic beef-protein medium of same use 10mL adds (90mL in 6 kinds of algae liquid respectively, all algae liquid all cultivates logarithmic phase) in contrast (1:9), the chlorophyll concentration measuring the algae liquid of all experimental group and control group after 4 days and 6 days is respectively cultivated in illumination box, calculate its molten algae efficiency.Three Duplicate Samples surveyed by each sample, are expressed as the form of mean+SD.
The algae kind used in experiment has 3 strains purchased from the aquatic institute in Wuhan algae kind storehouse, and other 3 strain screenings are from Taihu Lake water body.Experimental result shows, the fermented liquid of genus bacillus Lzh-5 has good algicidal effect (table 1) for most protokaryon algae kind.This result shows that the molten algae ability of genus bacillus Lzh-5 has broad spectrum, is the very potential algae-lysing bacterium of one.
Table 1. genus bacillus Lzh-5 is to the algicidal effect of 6 strain algae strains
Note: screen from the Mei Liangwan water body of Wuxi, Taihu Lake with the algae strain that asterisk (*) marks in table and obtain, all the other algae strains are all buy from CHINESE FRESHWATER algae kind storehouse.
Genus bacillus Lzh-5 to the algicidal effect of the main blue-green algae microcystic aeruginosa 9110 in the blue-green alga bloom of Taihu Lake as shown in Figure 1.Fig. 1 shows, after adding genus bacillus Lzh-5, microcystic aeruginosa 9110 cell concn prolongation in time and declining, and microcystic aeruginosa 9110 cell concn prolongation in time and rising in blank, microcystic aeruginosa 9110(> 91% nearly all when the 6th day) dead.The genus bacillus Lzh-5 starting point concentration wherein adding algae liquid is 1 × 10
8individual/mL.
The research of embodiment 5Lzh-5 bacterial strain algicidal mode
Genus bacillus Lzh-5 fermented liquid is made acetic acid ethyl acetate extract according to the method described in step 3, after centrifugal dryer evaporate to dryness, for subsequent use after dissolving with sterilized water.Finally same for sterilizing beef-protein medium centrifugal drying is concentrated in contrast.Extraction liquid obtained according to the method described above and the contrast of concentrated beef extract-peptone are added on the round scraps of paper (diameter 1cm) respectively, are positioned on BG11 solid plate that microcystic aeruginosa makes.Algae flat board is put into illumination box cultivation and within 2 days, is observed the formation of the algal control circle around the circle scraps of paper afterwards to judge its algicidal effect.
The making method of microcystic aeruginosa agar plate: pour appropriate (about 20mL) BG11 nutrient agar (agar ratio 1.5%) on every block flat board into, for subsequent use after it solidifies.Supernatant is discarded by after the centrifugal 20min of cultured for 300mL microcystic aeruginosa algae liquid 4000g, collect frustule and precipitate (1% agar in the BG11 soft agar solid medium after joining 100mL sterilizing, be positioned in 53 DEG C of water-baths and prevent from solidifying), pour the BG11 solid plate upper strata (every block flat board about pours 20mL into) made after shaking up into, put into after it solidifies illumination box cultivate and for subsequent use.
As shown in Figure 2, the fermented liquid extraction liquid of display genus bacillus Lzh-5 has good algicidal effect to experimental result, and this shows that this bacterial strain has the performance that the outer algicidal substances of secretion born of the same parents carries out molten algae.
The extraction purification of molten algae effective constituent in embodiment 6 genus bacillus Lzh-5 tunning
Utilize HPLC technology by the molten algae meta-bolites purifying of molten phycomycete Lzh-5, obtain purifying substance S-5A and S-5B that two kinds have molten algae function, concrete steps are as follows:
By the SupersilTM C18-EP semipreparative column of the extraction liquid of genus bacillus Lzh-5 fermented liquid by DIKMA company, water and methyl alcohol are that the HPLC technique means of moving phase carries out preliminary purification, obtain molten algae effective constituent.Then by DIKMA SupersilTM C18-EP analytical column, water and methyl alcohol are that the HPLC technique means of moving phase is further purified, and obtain two kinds of effective molten algae composition S-5A and S-5B, for Structural Identification.
The Identification of chemical structure of molten algae effective constituent in embodiment 7 genus bacillus Lzh-5 meta-bolites
LC-MS ﹑ GC-MG and NMR analysis (nuclear magnetic resonance spectroscopy) is utilized to have the chemical structure of the meta-bolites of molten algae function.
Sample S-5A after purifying is carried out LC-MS analysis, and the molecular ion peak obtaining sample S-5A is 155.0821m/z, and molecular weight analyte is 154.0742, and molecular formula is C
7h
10n
2o
2.
Sample S-5A after purifying is carried out GC-MS analysis, with the structural similitude of also [1, the 2-a] pyrazine-Isosorbide-5-Nitrae-diketone of hexahydropyrrolo in GC-MS database, similarity index > 850.
Sample S-5A is carried out hydrogen nuclear magnetic resonance spectrum analysis, obtains following result:
1H NMR(400MHz,D
2O)δ4.22(s,1H),4.06(dd,J=17.3,2.7Hz,1H),3.77(d,J=17.3Hz,1H),3.44(dd,J=8.7,4.8Hz,2H),2.41–2.12(m,1H),2.09–1.71(m,3H)。
The LC-MS ﹑ GC-MS of sample S-5A and NMR result are analyzed, the meta-bolites S-5A that can determine to have algicidal effect is hexahydropyrrolo also [1,2-a] pyrazine-Isosorbide-5-Nitrae-diketone, and its chemical structure is as shown below.
Sample S-5B after purifying is carried out LC-MS analysis, and the molecular ion peak obtaining sample S-5B is 197.1276m/z, and molecular weight analyte is 196.1212, and molecular formula is C
10h
16n
2o
2.
Sample S-5B after purifying is carried out GC-MS analysis, wherein the structural similitude of 3-sec.-propyl-hexahydropyrrolo also [1,2-a] pyrazine-Isosorbide-5-Nitrae-diketone in S-5B and known references.
Sample S-5B is carried out hydrogen nuclear magnetic resonance spectrum analysis, obtains following result:
1H NMR(400MHz,D
2O)δ4.20(m,1H),4.07(s,1H),3.47(dd,J=16.3,8.9Hz,1H),2.38(m,1H),2.26(d,J=4.7Hz,1H),1.97(s,1H),1.83(s,1H),0.98(d,J=7.2Hz,1H),0.76(d,J=6.9Hz,1H)。
Sample S-5B is carried out carbon-13 nmr spectra analysis, obtains following result:
13C NMR(101MHz,D
2O)δ172.23(s),167.12(m),60.34(s),58.72(m),45.20(s),28.89(s),28.10(s),21.55(s),17.95(s),15.06(s)。
The LC-MS ﹑ GC-MS of sample S-5B and NMR result are analyzed, the meta-bolites S-5B that can determine to have algicidal effect is 3-sec.-propyl-hexahydropyrrolo also [1,2-a] pyrazine-Isosorbide-5-Nitrae-diketone, and its chemical structure is as follows.
The molten algae effect research of embodiment 8 genus bacillus Lzh-5 molten algae meta-bolites hexahydropyrrolo also [1,2-a] pyrazine-Isosorbide-5-Nitrae-diketone and 3-sec.-propyl-hexahydropyrrolo also [1,2-a] pyrazine-Isosorbide-5-Nitrae-diketone
Use the method in the research of above-mentioned Lzh-5 bacterial strain algicidal mode, hexahydropyrrolo also [1 is found through experiment, 2-a] pyrazine-1,4-diketone and 3-sec.-propyl-hexahydropyrrolo also [1,2-a] pyrazine-1, the BG11 solid plate that 4-diketone is made at microcystic aeruginosa can form obvious algal control circle, and blank then can not form algal control circle (Fig. 3).
By hexahydropyrrolo also [1,2-a] pyrazine-1,4-diketone and 3-sec.-propyl-hexahydropyrrolo also [1,2-a] pyrazine-Isosorbide-5-Nitrae-diketone adds in blue-green algae and microcystic aeruginosa 9110 respectively, makes hexahydropyrrolo also [1,2-a] pyrazine-1, the concentration of 4-diketone and 3-sec.-propyl-hexahydropyrrolo also [1,2-a] pyrazine-Isosorbide-5-Nitrae-diketone forms a series respectively: 100 μ g/mL ﹑ 50 μ g/mL ﹑ 40 μ g/mL ﹑ 30 μ g/mL ﹑ 20 μ g/mL ﹑ 10 μ g/mL and 5 μ g/mL.The microcystic aeruginosa 9110 that will add hexahydropyrrolo also [1,2-a] pyrazine-Isosorbide-5-Nitrae-diketone and 3-sec.-propyl-hexahydropyrrolo also [1,2-a] pyrazine-Isosorbide-5-Nitrae-diketone is put into illumination box and is cultivated after 24 hours and measure molten algae rate.Experimental result shows, hexahydropyrrolo is [1,2-a] pyrazine-Isosorbide-5-Nitrae-diketone and 3-sec.-propyl-hexahydropyrrolo also [1 also, 2-a] the Lethal Dose 50 LD50 of pyrazine-Isosorbide-5-Nitrae-diketone to microcystic aeruginosa 9110 and synechococcus BN60 be respectively 5.7 μ g/mL and 19.4 μ g/mL (Fig. 4).The meta-bolites hexahydropyrrolo that this result shows genus bacillus Lzh-5 also [1,2-a] pyrazine-Isosorbide-5-Nitrae-diketone and 3-sec.-propyl-hexahydropyrrolo also [1,2-a] pyrazine-Isosorbide-5-Nitrae-diketone be the material of highly effective control blue-green algae.
Add in Fig. 4 different concns algicidal substances hexahydropyrrolo also [1,2-a] pyrazine-Isosorbide-5-Nitrae-diketone and 3-sec.-propyl-hexahydropyrrolo also [1,2-a] pyrazine-Isosorbide-5-Nitrae-diketone time microcystic aeruginosa 9110 survival rate change curve.
The algicidal effect of genus bacillus Lzh-5 of the present invention has broad spectrum.Its fermented liquid preparation method is simple, and preparation cycle is short.The fermented liquid of genus bacillus Lzh-5 has good algicidal effect to the microcystic aeruginosa 9110 in Taihu Lake and the algae BN35 etc. that quivers.Genus bacillus Lzh-5 fermented liquid and acetic acid ethyl acetate extract all can be used for the control of blue-green alga bloom or other micro-algae, and algicidal effect is better.Wherein, genus bacillus Lzh-5 fermented liquid reaches 91.2 ± 6.3% to 6 of microcystic aeruginosa 9110 days molten algae rates.The meta-bolites hexahydropyrrolo of genus bacillus Lzh-5 also [1,2-a] pyrazine-1,4-diketone and 3-sec.-propyl-hexahydropyrrolo also [1,2-a] pyrazine-1,4-diketone has good lethal effect to the main blue-green algae microcystic aeruginosa 9110 in the blue-green alga bloom of Taihu Lake, can be used for research and development and the production of novel algicide, be finally applied to the control of lake blue algae wawter bloom.
More than describe preferred embodiment of the present invention in detail.Should be appreciated that those of ordinary skill in the art just design according to the present invention can make many modifications and variations without the need to creative work.Therefore, all technician in the art, all should by the determined protection domain of claims under this invention's idea on the basis of existing technology by the available technical scheme of logical analysis, reasoning, or a limited experiment.
Claims (10)
1. a strain has genus bacillus (Bacillus sp.) Lzh-5 of molten algae activity, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preserving number CGMCC No.8282, and preservation date is on September 27th, 2013.
2. genus bacillus Lzh-5 as claimed in claim 1 is controlling the application in blue-green alga bloom.
3. the tunning of genus bacillus Lzh-5 as claimed in claim 1 is controlling the application in blue-green alga bloom.
4. a compound with structure shown in structural formula (II) is controlling the application in blue-green alga bloom.
5. a molten phycomycete agent, is characterized in that, comprises genus bacillus according to claim 1 (Bacillus sp.) Lzh-5 in described molten phycomycete agent.
6. a molten algae medicament, is characterized in that, comprise the tunning of genus bacillus according to claim 1 (Bacillus sp.) Lzh-5 in described molten algae medicament, described tunning is fermented liquid, concentrate of fermentation liquid or fermentation broth extract.
7. control a method for blue-green alga bloom, comprise the steps:
1), ferment genus bacillus Lzh-5 according to claim 1;
2), extractive fermentation liquid;
3), extraction liquid evaporate to dryness obtains crude extract;
4), crude extract water-soluble after for controlling blue-green alga bloom.
8. method as claimed in claim 7, wherein step 1) in, fermentation condition is, genus bacillus Lzh-5 is inoculated in the sterilizing beef-protein medium of pH 7.0,28 DEG C, 200rpm envrionment conditions bottom fermentation 48h, obtain described genus bacillus Lzh-5 fermented liquid.
9. method, wherein step 2 as claimed in claim 7) in, extraction agent is ethyl acetate.
10. method, wherein step 2 as claimed in claim 9) in, the volume ratio of ethyl acetate and fermented liquid is 1:1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310683393.6A CN103695342B (en) | 2013-12-12 | 2013-12-12 | One strain has genus bacillus and the application thereof of molten algae activity |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310683393.6A CN103695342B (en) | 2013-12-12 | 2013-12-12 | One strain has genus bacillus and the application thereof of molten algae activity |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103695342A CN103695342A (en) | 2014-04-02 |
| CN103695342B true CN103695342B (en) | 2015-08-12 |
Family
ID=50357006
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310683393.6A Expired - Fee Related CN103695342B (en) | 2013-12-12 | 2013-12-12 | One strain has genus bacillus and the application thereof of molten algae activity |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103695342B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104046581B (en) * | 2014-06-04 | 2016-10-19 | 上海交通大学 | One strain molten algae Chryseobacterium sp and the application in blue-green alga bloom control thereof |
| CN107177538B (en) * | 2017-07-19 | 2019-12-06 | 扬州大学 | Marine bacillus cereus and application thereof in preventing and controlling anabaena flos-aquae |
| CN111826318B (en) * | 2020-07-30 | 2022-03-29 | 中冶华天工程技术有限公司 | Microcystis aeruginosa dissolving bacterium and application thereof |
| CN115161231B (en) * | 2022-06-23 | 2023-03-31 | 轩凯生物科技(滁州)有限公司 | Bacillus coagulans with algae dissolving function and application thereof |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101280278A (en) * | 2008-05-20 | 2008-10-08 | 云南大学 | Separation method and application of algicadal bacteria |
| CN102154170B (en) * | 2011-01-11 | 2012-11-07 | 常州大学 | Microcystin degrading strain and method for degrading MC-LR (microcystins-LR) by same |
| CN102206605B (en) * | 2011-04-29 | 2013-01-16 | 上海交通大学 | Exiguobacterium sp. with alga-lysing activity and application thereof in cyanobacterial bloom control |
| CN102839139A (en) * | 2012-08-28 | 2012-12-26 | 安徽联大生物环保科技有限公司 | Solid fermentation and application of bacillus algae dissolving bacteria |
| CN103409348B (en) * | 2013-07-25 | 2014-07-23 | 南京厚稷生物科技有限公司 | Bacillus pumilus with efficient alga-lysing activity and application thereof |
| CN103509744B (en) * | 2013-09-11 | 2015-07-15 | 上海交通大学 | Algicidal Shewanella and application thereof in controlling blue algae water bloom |
-
2013
- 2013-12-12 CN CN201310683393.6A patent/CN103695342B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN103695342A (en) | 2014-04-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103667135B (en) | One strain Stenotrophomonas and application thereof | |
| CN102311981B (en) | Method for preparing and purifying prodigiosin | |
| CN103695342B (en) | One strain has genus bacillus and the application thereof of molten algae activity | |
| CN102154170A (en) | Microcystin degrading strain and method for degrading MC-LR (microcystins-LR) by same | |
| CN102174051A (en) | Strong algae activity inhibiting compound as well as preparation method and application thereof | |
| CN103937726A (en) | Alga-lysing pseudomonas aeruginosa and application thereof | |
| CN105950500A (en) | Algae-lysing aeromonas sp. and application thereof in controlling cyanobacterial blooms | |
| CN102659912B (en) | Oxygen-rich disesquiterpenes compound, and preparation method and application thereof | |
| CN103074279B (en) | Paenibacillus sp. PN-S435 and application thereof in microcystins (MCs) degradation | |
| CN103509744B (en) | Algicidal Shewanella and application thereof in controlling blue algae water bloom | |
| CN102875551A (en) | Chloro alkaloid compound and preparation and application thereof | |
| CN103509745B (en) | One strain Stenotrophomonas and the application in blue-green alga bloom controls thereof | |
| CN103352010B (en) | Bacillus cereus strain CZBC1 capable of dissolving pond oscillatoria, and its application | |
| CN103952359A (en) | Brevundimonas sp. and application thereof | |
| CN102206605B (en) | Exiguobacterium sp. with alga-lysing activity and application thereof in cyanobacterial bloom control | |
| CN104804071B (en) | A kind of depside peptides and its preparation method and application | |
| CN102659547B (en) | Ophiobolin sesterterpene compound and preparation and application thereof | |
| CN104046581B (en) | One strain molten algae Chryseobacterium sp and the application in blue-green alga bloom control thereof | |
| CN102965310B (en) | Shinella sp. and application thereof to micro-biologically degrading acetaminophen | |
| CN102786528B (en) | Polyoxybiotic alkali compound as well as preparation method and application thereof | |
| CN105385642A (en) | Brevundimonas and application thereof | |
| CN104130960B (en) | One strain Exiguobacterium sp and the application in controlling blue-green alga bloom of effective molten algae composition thereof | |
| CN102701935B (en) | Tetranuclear diterpenoids as well as preparation and application thereof | |
| CN103045488A (en) | Beauveria felina AS-70 strain and application thereof | |
| CN105802872A (en) | Fluorescent pseudomonas, method for producing phenazine amide and application of phenazine amide |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20150812 Termination date: 20171212 |