CN104726369A - Raoultella ornithinolytica and applications thereof - Google Patents
Raoultella ornithinolytica and applications thereof Download PDFInfo
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Abstract
The invention discloses a raoultella ornithinolytica P1-A-19, which is prepared by carrying out enrichment, separation, purification and screening on sludge produced by sewage treatment plants. The raoultella ornithinolytica P1-A-19 is characterized in that the strain shows an electricity-production capacity when applied to a microbial fuel cell, and also has a capacity of converting baccatin III so as to generate 10-deacetyl baccatin III (10-DAB), and the Preservation Number of the strain is CGMCC No. 10267. The raoultella ornithinolytica P1-A-19 disclosed by the invention has a great significance for the enrichment of diversity of electricity-production microorganisms and the excavation of more microbial strains with high electrochemical activity; and microbial strains for 10-DAB biosynthesis are further widened, thereby providing new materials and new methods for the development of new taxane compound synthesis approaches.
Description
Technical field
The invention belongs to microorganism field, relate to a kind of the new strains solution ornithine Raoul bacterium and the application thereof that there is electricity generation ability and baccatin III can be transformed generation 10-DAB.
Background technology
1. electrogenesis microorganism
Microbiological fuel cell (Microbial Fuel Cells, MFCs) be a kind ofly utilize microorganism to do catalyst oxidation organism and direct new device chemical energy being become electric energy, it is in conjunction with fuel cell and biology techniques means, achieve biological chemistry can with the direct conversion of electric energy, be a kind of clean sustainable energy, become the focus of new energy development gradually.
Electrogenesis microorganism (Electricigens) refers to that those can under anaerobic complete oxidation organism, the electronics that oxidation of organic compounds obtains is delivered to extracellular by electron transport chain, directly or indirectly through the microorganism of medium (Mediator) by generation current on electron transmission to electrode, microorganism obtains energy support growth in electron transfer process simultaneously.Electrogenesis microorganism, as biological catalyst important in microbiological fuel cell, directly affects efficiency of fuel cell generation.Therefore excavate the microorganism with this function for the diversity enriching electrogenesis microorganism more, improve efficiency of fuel cell generation significant.
The microorganism that can run MFCs electrogenesis under without amboceptor condition reported at present comprises bacterium class and Mycophyta.Wherein, bacterium class electrogenesis microorganism mainly comprises genus Shewanella (Shewanella), ground Bacillaceae (Geobacter), pseudomonas Pseudomonas (Pseudomonas), arc Pseudomonas (Arcobacter), the electrogenesis bacterium of hydrogen-producing bacteria family is (as clostridium butylicum (Clostridium butyricum), enteroaerogen (Enterobacteraerogenes), Fe3+ reduction rhodospirillum (Rhodoferax ferrireducens), human pallid bacillus (Ochrobactrumanthropi), cryophilic bacteria (Geopsychrobacter electrodiphilus), Klebsiella pneumoniae (Klebsiella pneumoniae L17) etc.The electrogenesis microorganism of Mycophyta mainly comprises Hansenula anomala (Hansenula anomala), swamp Rhodopseudomonas (Rhodopseudomonas palustris), chlorella (Chlorellavulgaris) etc.But not yet find that the bacterial strain of Raoul Pseudomonas has electricity generation performance so far.
2.10-DAB preparation
Docetaxel is the Typical Representative of the taxane anti-tumor medicament through structural modification, has the anti-tumor activity of high-efficiency broad spectrum, and oneself is through becoming the first-line drug of the oncotherapy such as ovarian cancer, mammary cancer at present.And 10-DAB is the important precursor of synthesis anticancer drug docetaxel.
10-DAB is distributed widely in the leaf of Chinese yew, root, skin, limb.Highly finished product could be obtained after traditional extracting method needs repeatedly chromatography, gradient elution, repeatedly recrystallization.This not only causes a large amount of losses of 10-DAB, reduces yield, and also uses sherwood oil inflammable in a large number and virose acetonitrile in process, causes detrimentally affect to environment.And utilizing microbiological transformation technology baccatin III can be transformed generation 10-DAB, it is high that this process has specificity, and by product generates few, and reaction conditions is gentle, advantages of environment protection.
What reported at present can take baccatin III as substrate, transforms the microorganism generating 10-DAB and mainly contains: white group Nocardia bacteria (Nocardioides albus), yellow class Nocardia bacteria (Nocardioides luteus), Pseudomonas aeruginosa (Pseudomonas aeruginosa), Pseudomonas straminea (Pseudomonas straminea), pantoea agglomerans (Pantoea agglomerans), rhodococcus (Rhodococcus sp.), catarrhalis (Moraxella sp.) etc.So far not yet find that the bacterial strain of Raoul Pseudomonas possesses and baccatin III is transformed the ability generating 10-DAB.
Summary of the invention
The object of the present invention is to provide a strain to have electricity generation ability and baccatin III can be transformed the new strains of generation 10-DAB and study it in microbiological fuel cell and the application prepared in 10-DAB.
The present invention is achieved by the following technical solutions:
One strain solution ornithine Raoul bacterium (Raoultella ornithinolytica) P1-A-19, bacterial strain deposit number: CGMCC No.10267, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), address: No. 1, North Star West Road, Chaoyang District, city of BeiJing, China institute, preservation date: on December 30th, 2014.
And described bacterial strain has electricity generation ability and baccatin III is transformed the ability generating 10-DAB,
And described bacterial strain is applied in microbiological fuel cell and shows electricity generation ability.
And described bacterial strain contains or expresses C-10 deacetylase.
Have electricity generation ability and baccatin III transformed the screening method of bacterial strain generating 10-DAB, step is as follows:
(1) enrichment
Mud between sludge condensation to be screened through simply eluriate and precipitation process after, remove the clear water on the inorganization throw out of bottom and top layer, get intermediate aqueous rate about 95% mud as inoculum, add together with mud nutritive medium in the reactor of microbiological fuel cell, continuous operation 10-20d, the voltage at pull-up resistor two ends is stablized gradually, now, anode is formed macroscopic thick microbial film;
(2) separation and purification
With transfering loop, the whole thalline on microbial film are scraped, be suspended in sterilized water and be prepared into uniform bacteria suspension, be coated on glucose as on the PBBM solid medium of carbon source, 30 DEG C of Anaerobic culturel 3-5d, the single bacterium colony with different shape grown is chosen respectively, the line abstraction and purification carried out under above-mentioned identical substratum and culture condition is repeatedly cultivated, and finally obtains multiple purebred strain isolated, is numbered and names;
(3) screen
1. will be separated the inoculation that obtains in anolyte I, the inoculation liquid obtained injects the reactor of microbiological fuel cell, and reactor and external resistance and data collector connect, and run under room temperature;
2. thalline collection obtained, after toluene break process, adds substrate baccatin III, and 37 DEG C of 200r/min transform 3d, are identified converted product by TLC, HPLC, LC-MS.
Solution ornithine Raoul bacterium P1-A-19 according to claim 1 is used for the application carrying out electrogenesis in microbiological fuel cell.
Separate ornithine Raoul bacterium (Raoultella ornithinolytica) P1-A-19 and make the application in microbiological fuel cell.
Separate the application of ornithine Raoul bacterium (Raoultella ornithinolytica) P1-A-19 in preparation 10-DAB.
Separate the method that ornithine Raoul bacterium (Raoultella ornithinolytica) P1-A-19 prepares 10-DAB, bacterial strain collection obtained, after toluene break process, adds substrate baccatin III, 30-40 DEG C, 180-250r/min transforms 3-8d, obtains 10-DAB.
Advantage of the present invention and positively effect as follows:
The present invention to have electricity generation ability as preliminary screening condition in MFCs; 10-DAB is generated as multiple grating part to transform baccatin III; through the strict restriction of these two steps; screening obtains a strain bacterial strain Raoultella ornithinolytica P1-A-19; strict restriction screening through this two steps condition obtains has electricity generation performance and the bacterial strain containing C-10 deacetylase; bacterial strain is carried out the experiment of MFCs electrogenesis and transform baccatin III experiment; find that bacterial strain can produce the voltage of 132mV, and baccatin III can be transformed generation 10-DAB III.
The present invention is for the diversity enriching electrogenesis microorganism, and to excavate the microbial strains with high electrochemical activity significant more; Also widened for the biosynthetic microorganism strains of 10-DAB further, for the route of synthesis developing new taxane compounds provides novel material and novel method simultaneously.
Accompanying drawing explanation
Fig. 1 is the colonial morphology figure of bacterial strain Raoultella ornithinolytica P1-A-19 of the present invention;
Fig. 2 is the gramstaining result of bacterial strain Raoultella ornithinolytica P1-A-19 of the present invention;
Fig. 3 is the phylogeny tree graph of bacterial strain Raoultella ornithinolytica P1-A-19 of the present invention;
Fig. 4 is the voltage-time curve figure of bacterial strain Raoultella ornithinolytica P1-A-19 of the present invention when being applied to MFCs;
Fig. 5 is the TLC result that bacterial strain Raoultella ornithinolytica P1-A-19 of the present invention transforms baccatin III, 1: baccatin III standard substance (0.3mg/mL), 2:10-DAB standard substance (0.1mg/mL), 3: phage control, 4: conversion fluid sample;
Fig. 6 is the HPLC result that bacterial strain Raoultella ornithinolytica P1-A-19 of the present invention transforms baccatin III, Fig. 6 A: the hybrid standard product of baccatin III (0.3mg/mL) and 10-DAB (0.1mg/mL), Fig. 6 B: conversion fluid sample;
Fig. 7 is the LC-MS figure that bacterial strain Raoultella ornithinolytica P1-A-19 of the present invention transforms the converted product of baccatin III, Fig. 7 A:TOF MS negative ion scan pattern, Fig. 7 B:TOF MS positive ion scan pattern.
Embodiment
Below by specific embodiment, the invention will be further described, and following examples are descriptive, is not determinate, can not limit protection scope of the present invention with this.
Content of the present invention comprises following content:
One, be placed in single chamber type microbiological fuel cell by the mud between being concentrated by TEDA in Tianjin's sludge of sewage treatment plant to run, with the microbial film of enrichment on the anode be observed visually for research object, the microorganism utilizing educable method separation and purification to obtain having electrochemical activity is purebred.
Two, analyze by running the electricity generation performance of MFCs to strain isolated P1-A-19.
Three, by TLC, HPLC, LC-MS, the ability that strain isolated P1-A-19 transforms baccatin III generation 10-DAB is studied.
Specifically be discussed below:
One, the separation screening qualification key step of bacterial strain is as follows:
(1) separation screening of strain isolated
Mud between sludge condensation to be screened through simply eluriate and precipitation process after, remove the clear water on the inorganization throw out of bottom and top layer, get intermediate aqueous rate about 95% mud as inoculum, add together with mud nutritive medium in the reactor of microbiological fuel cell, continuous operation 10-20d, the voltage at pull-up resistor two ends is stablized gradually, now, anode is formed macroscopic thick microbial film;
With transfering loop, the microorganism on microbial film is scraped, be suspended in sterilized water and be prepared into uniform bacteria suspension, be coated on glucose as on the PBBM solid medium of carbon source, 30 DEG C of Anaerobic culturel 3d, the single bacterium colony with different shape grown is chosen respectively, under identical substratum and culture condition, carries out three rides, 30 DEG C of Anaerobic culturel 3d, after separating for several times purifying, obtain purebred strain isolated, be numbered and name.
(2) qualification of strain isolated P1-A-19
Solution ornithine Raoul bacterium (Raoultella ornithinolytica) P1-A-19 of the present invention has following physiology and morphology biochemical characteristic:
A. the morphological specificity of thalline: Gram-negative bacteria, shaft-like;
B. the morphological specificity of bacterium colony: after nutrient agar plate cultivates 24h, colonial morphology is circular, easy picking, opaque, surface wettability, the white colony of neat in edge, and diameter is about 1-1.5mm;
C. main physio-biochemical characteristics: indoles produces experiment and urease experimental result is positive, methyl red experiment, hydrogen sulfide experiment and oxydase experiment are negative.
The above results shows, the morphological specificity of bacterial strain P1-A-19 of the present invention is all consistent with the solution ornithine Raoul bacterium (Raoultella ornithinolytica) of bibliographical information with physio-biochemical characteristics.
D. solution ornithine Raoul bacterium (Raoultella ornithinolytica) P1-A-19 molecular classification status of the present invention:
Adopt Easypure
tMgenomic DNA kit test kit (Tian Gen bio tech ltd, Beijing) extract bacterial strain P1-A-19 STb gene and with for template, pcr amplification is carried out to 16S rDNA gene order.Primer pair is universal primer 27F and 1492R.PCR primer direct Sequencing, sequencing result submits NCBIGenBank (National Center ofBiotechnology Information) to, carries out similarity retrieval and tetraploid rice with BLAST.Utilize MEGA5 software, use Neighbor-Joining method, adopt Kimura2-parameter calibration model, 1000 phylogenetic tree constructions of booting.
Result shows, the homology of this bacterial strain and Raoultella ornithinolytica ATCC 318989 (NR 114502.1) is the highest, reach 99%, strain isolated P1-A-19 and Raoultella ornithinolytica ATCC31898 is in the same branch of phylogenetic tree, and sibship is the most close.
Comprehensive the above results, bacterial strain P1-A-19 of the present invention is accredited as and separates ornithine Raoul bacterium (Raoultellaornithinolytica).
Two, the analysis of electricity generation performance
(1) structure of microbiological fuel cell and assembling
What adopt is single chamber type MFCs reactor.It is only made up of a pond body, and negative electrode one side is exposed in the middle of air, and the useful area of anode and cathode is 7cm
2.Main body be one long for 2.0cm, cross-sectional diameter is the synthetic glass cylinder of 3.0cm, and useful volume is 14mL.Anode and negative electrode are placed in respectively the both sides of MFCs main body, the sealing of blend rubber circle is fixing.Anode one end plexiglass cover covers, and in order to ensure that negative electrode contacts with air, the lid of negative electrode one end is opening, and negative electrode carbon cloth carries platinum.Anodic-cathodic is all connected with titanium silk to be derived electronics or imports, and last whole device screw and leading screw are tightened fixing, and MFCs reactor has built.
(2) start-up and operation of MFCs
Strain isolated P1-A-19 is inoculated in anolyte I, and the inoculation liquid that obtains injects MFCs reactor, and reactor and external resistance (not having specified otherwise external resistance resistance to be fixed on 1000 Ω) and data collector connect, and run under room temperature.
Periodic replacement inoculation liquid (being namely vaccinated with the anolyte of corresponding strain isolated thalline) is needed in operational process, general 24h changes once, first the former inoculation liquid in MFCs is taken out during each replacing, then add fresh inoculation liquid, question response device occurs that the stabilized voltage of continuous three can be thought and starts successfully.Start successfully and enter the commencement of commercial operation phase, when the output voltage of monitoring is lower than 50mV, only needs to change anolyte, carry out the mensuration of polarization curve and power density curve when output voltage again reaches the highest and tends towards stability, stop MFCs running afterwards.Utilize PISO-813 type data collecting system to carry out real-time online data monitoring and record to voltage in whole process, get the mean value in 30min during analytical data, sampling precision is 0.001V.
(3) the Electrochemical Characterization parameter of MFCs
The electric current that the small-sized MFCs in one, laboratory produces is very little, is difficult to directly measure, but is calculated by the voltage measured in external resistance.The voltage that in this paper, all MFCs produce all is undertaken gathering and recording by PISO-813 type data collecting system.The maximum voltage that MFCs produces is open circuit voltage, can record in the infinitely-great situation of external resistance.
Test as stated above, result shows, shows electrochemical activity when bacterial strain P1-A-19 is applied to microbiological fuel cell, and its maximum voltage produced is 132mV.
Three, by TLC, HPLC, LC-MS, the ability that strain isolated P1-A-19 transforms baccatin III generation 10-DAB is studied.
(1) be inoculated into by the thalline P1-A-19 on inclined-plane in the 250mL triangular flask that 50mL LB liquid nutrient medium is housed, 30 DEG C of 200r/min cultivate 2d.Medium centrifugal (7000r/min, 4 DEG C, 10min), removes supernatant liquor, and collecting precipitation wets bacterium mud, and measures bacterium shale amount, stand-by.Adopt O for toluene crush method, namely every 2g wets in bacterium mud and adds 0.04g NaHCO
3with 8mL toluene, 37 DEG C of 200r/min shaking tables vibrate 2.5h, remove toluene, collect thalline and are used for transforming.
(2) collect thalline 6g and be placed in test tube, add the baccatin III methanol solution 2mL of 1.5mg/mL, then add 50mmol/L phosphate buffered saline buffer (pH 7.2) 8mL, now the final concentration of substrate is 0.3mg/mL.Test tube is placed in shaking table, 37 DEG C, 200r/min transforms 3d.Be used for termination reaction by adding 5mL chloroform in transformation system in experiment, afterwards, test tube continues at 37 DEG C, and chloroform organic phase, to extract converted product, finally moves in phial, volatilizes solvent under being placed in room temperature by the 3h that vibrates under 200r/min condition.In experiment, identical thalline is suspended in 10mL 50mmol/L phosphate buffered saline buffer (pH 7.2) as blank.Utilize silica-gel plate to carry out TLC separation to converted product, developping agent is acetone: sherwood oil=2:3 (v/v), and developer is sulfuric acid: ethanol=1:9 (v/v).And utilize HPLC, LC-MS to identify converted product.
(3) efficient liquid phase chromatographic analysis (HPLC) concrete grammar: add 800 μ L methyl alcohol in sample, after ultra-sonic oscillation mixing, obtains need testing solution and carries out HPLC analysis after 0.45 μm of filtering with microporous membrane.Chromatographic condition: C18 chromatographic column (250mm × 4.6mm, 5 μm); Moving phase: methyl alcohol: acetonitrile: water (10:33:57, v/v/v); Determined wavelength: 227nm; Flow velocity: 0.8mL/min; Column temperature: room temperature; Sample size: 10 μ L.
(4) high performance liquid chromatography mass spectrometry analyzes (LC-MS) concrete grammar: chromatographic condition: C18 chromatographic column (100mm × 2.1mm); Moving phase: 0.1%FA-acetonitrile, 0-10min, 5-95% acetonitrile; Flow velocity: 0.4mL/min; Column temperature: 40 DEG C, sample size 10 μ L.Mass Spectrometry Conditions: ionizer: electron spray ionisation (ESI); Detecting pattern: multiple-reaction monitoring; Capillary voltage: 3kV; Desolventizing temperature degree: 400 DEG C; Desolventizing airshed: 800L/h; Taper hole airshed: 30L/h; Ion source temperature: 350 DEG C; Impinging air flows speed 0.12mL/min.
(5) according to said method test, found that, on TLC plate bacterial strain P1-A-19 conversion fluid sample in there are three spot A (R
fvalue 0.31), B (R
fvalue 0.25), C (R
fvalue 0.12), the wherein color of spot C and 10-DAB standard substance and Rf value R
fidentical.There is obvious detached peaks in the HPLC figure of conversion fluid sample, can tentatively judge in conversion fluid sample containing 10-DAB near 10-DAB retention time.And in the TOF MS negative ion spectrogram of converted product, observe the detached peaks that molecular weight is 544.2256, just caing be compared to the few ethanoyl of substrate baccatin III, and be C with molecular formula
29h
36o
1010-DAB molecular weight consistent.
The result that comprehensive TLC, HPLC, LC-MS analyze is known, and strain isolated has the ability transforming baccatin III generation 10-DAB.
10-DAB is distributed widely in the leaf of Chinese yew, root, skin, limb.Highly finished product could be obtained after traditional extracting method needs repeatedly chromatography, gradient elution, repeatedly recrystallization.This not only causes a large amount of losses of 10-DAB, reduces yield, and also uses sherwood oil inflammable in a large number and virose acetonitrile in process, causes detrimentally affect to environment.
Utilize bacterial strain provided by the invention baccatin III one step can be transformed and generate 10-DAB, it is high that this microbial conversion process has specificity, and by product generates few, and reaction conditions is gentle, advantages of environment protection.
The base sequence of the 16S rDNA mensuration of bacterial strain Raoultella ornithinolytica P1-A-19 of the present invention; TTAAGCTACCTACTTCTTTTGCAACCCACTCCCATGGTGTGACGGGCGGTGTGTACAAGGCCCGGGAACGTATTCACCGTAGCATTCTGATCTACGATTACTAGCGATTCCGACTTCATGGAGTCGAGTTGCAGACTCCAATCCGGACTACGACATACTTTATGAGGTCCGCTTGCTCTCGCGAGGTCGCTTCTCTTTGTATATGCCATTGTAGCACGTGTGTAGCCCTACTCGTAAGGGCCATGATGACTTGACGTCATCCCCACCTTCCTCCAGTTTATCACTGGCAGTCTCCTTTGAGTTCCCGACCGAATCGCTGGCAACAAAGGATAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATTTCACAACACGAGCTGACGACAGCCATGCAGCACCTGTCTCAGAGTTCCCGAAGGCACCAAAGCATCTCTGCTAAGTTCTCTGGATGTCAAGAGTAGGTAAGGTTCTTCGCGTTGCATCGAATTAAACCACATGCTCCACCGCTTGTGCGGGCCCCCGTCAATTCATTTGAGTTTTAACCTTGCGGCCGTACTCCCCAGGCGGTCGACTTAACGCGTTAGCTCCGGAAGCCACTCCTCAAGGGAACAACCTCCAAGTCGACATCGTTTACAGCGTGGACTACCAGGGTATCTAATCCTGTTTGCTCCCCACGCTTTCGCACCTGAGCGTCAGTCTTTGTCCAGGGGGCCGCCTTCGCCACCGGTATTCCTCCAGATCTCTACGCATTTCACCGCTACACCTGGAATTCTACCCCCCTCTACAAGACTCAAGCCTGCCAGTTTCAAATGCAGTTCCCAGGTTGAGCCCGGGGATTTCACATCTGACTTAACAGACCGCCTGCGTGCGCTTTACGCCCAGTAATTCCGATTAACGCTTGCACCCTCCGTATTACCGCGGCTGCTGGCACGGAGTTAGCCGGTGCTTCTTCTGCGAGTAACGTCAATCACTAAGGTTATTAACCTTAACGCCTTCCTCCTCGCTGAAAGTACTTTACAACCCGAAGGCCTTCTTCATACACGCGGCATGGCTGCATCAGGCTTGCGCCCATTGTGCAATATTCCCCACTGCTGCCTCCCGTAGGAGTCTGGACCGTGTCTCAGTTCCAGTGTGGCTGGTCATCCTCTCAGACCAGCTAGGGATCGTCGCCTAGGTGAGCCATTACCCCACCTACTAGCTAATCCCATCTGGGCACATCTGATGGCATGAGGCCCGAAGGTCCCCCACTTTGGTCTTGCGACGTTATGCGGTATTAGCTACCGTTTCCAGTAGTTATCCCCCTCCATCAGGCAGTTTCCCAGACATTACTCACCCGTCCGCCGCTCGTCACCCGAGAGCAAGCTCTCTGTGCTACCGCTCGA。
Claims (9)
1. a strain solution ornithine Raoul bacterium P1-A-19, it is characterized in that: bacterial strain deposit number: CGMCCNo.10267, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, address: No. 1, North Star West Road, Chaoyang District, city of BeiJing, China institute, preservation date: on December 30th, 2014, classification number is: Raoultellaornithinolytica.
2. solution ornithine Raoul bacterium P1-A-19 according to claim 1, is characterized in that: described bacterial strain has electricity generation ability.
3. solution ornithine Raoul bacterium P1-A-19 according to claim 1, is characterized in that: described bacterial strain contains or expresses C-10 deacetylase.
4. solution ornithine Raoul bacterium P1-A-19 according to claim 1, is characterized in that: baccatin III can transform and generate 10-DAB by described bacterial strain.
5. there is electricity generation ability and by a screening method for the bacterial strain of baccatin III conversion generation 10-DAB, it is characterized in that: step is as follows:
(1) enrichment
Mud between sludge condensation to be screened through simply eluriate and precipitation process after, remove the clear water on the inorganization throw out of bottom and top layer, get intermediate aqueous rate about 95% mud as inoculum, add together with mud nutritive medium in the reactor of microbiological fuel cell, continuous operation 10-20d, the voltage at pull-up resistor two ends is stablized gradually, now, anode is formed macroscopic thick microbial film;
(2) separation and purification
With transfering loop, the whole thalline on microbial film are scraped, be suspended in sterilized water and be prepared into uniform bacteria suspension, be coated on glucose as on the PBBM solid medium of carbon source, 30 DEG C of Anaerobic culturel 3-5d, the single bacterium colony with different shape grown is chosen respectively, the line abstraction and purification carried out under above-mentioned identical substratum and culture condition is repeatedly cultivated, and finally obtains multiple purebred strain isolated, is numbered and names;
(3) screen
1. will be separated the inoculation that obtains in anolyte I, the inoculation liquid obtained injects the reactor of microbiological fuel cell, and reactor and external resistance and data collector connect, and run under room temperature;
2. thalline collection obtained, after toluene break process, adds substrate baccatin III, and 37 DEG C of 200r/min transform 3d, are identified converted product by TLC, HPLC, LC-MS.
6. solution ornithine Raoul bacterium P1-A-19 according to claim 1 is used for the application of electrogenesis in microbiological fuel cell.
7. bacterial strain according to claim 1 is making the application in microbiological fuel cell.
8. the application of bacterial strain according to claim 1 in preparation 10-DAB.
9. bacterial strain according to claim 1 prepares the method for 10-DAB, it is characterized in that: bacterial strain collection obtained is after toluene break process, and add substrate baccatin III, 30-40 DEG C, 180-250r/min transform 3-8d, obtains 10-DAB.
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| CN108410758A (en) * | 2018-03-05 | 2018-08-17 | 南京理工大学 | Triazole degradation bacteria and its application in wastewater treatment containing triazole |
| CN108410768A (en) * | 2018-03-19 | 2018-08-17 | 陕西科技大学 | A kind of method of fermentation and hydrogen production bacterium and prophylactic activity sludge calcification |
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