CN106047754A - Method for improving power generation capacity of clostridium beijerinckii and application of clostridium beijerinckii - Google Patents
Method for improving power generation capacity of clostridium beijerinckii and application of clostridium beijerinckii Download PDFInfo
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- CN106047754A CN106047754A CN201610399488.9A CN201610399488A CN106047754A CN 106047754 A CN106047754 A CN 106047754A CN 201610399488 A CN201610399488 A CN 201610399488A CN 106047754 A CN106047754 A CN 106047754A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
- C07K14/33—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Clostridium (G)
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- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01M—PROCESSES OR MEANS, e.g. BATTERIES, FOR THE DIRECT CONVERSION OF CHEMICAL ENERGY INTO ELECTRICAL ENERGY
- H01M8/00—Fuel cells; Manufacture thereof
- H01M8/16—Biochemical fuel cells, i.e. cells in which microorganisms function as catalysts
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E60/00—Enabling technologies; Technologies with a potential or indirect contribution to GHG emissions mitigation
- Y02E60/30—Hydrogen technology
- Y02E60/50—Fuel cells
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P70/00—Climate change mitigation technologies in the production process for final industrial or consumer products
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Abstract
The invention discloses a method for improving power generation capacity of clostridium beijerinckii and an application of clostridium beijerinckii. According to the method, genes of encoded membrane protein Cbei_3304 is subjected to insertional inactivation, so that the genes in the clostridium beijerinckii cannot be normally expressed. An obtained recombinant strain glucose seed solution with high power generation capacity has the maximum output voltage up to 409 mV and the maximum output power density up to 144.4 mW/m<2> when methyl viologen is taken as an electron mediator. The process is simple, operation is convenient, no pollution is caused, the cost is low, the fuel utilization range is wider, the electron recovery rate is relatively high, and energy crisis is relieved; high-yield clostridium beijerinckii is a good strain suitable for research and applications of microbial fuel cells.
Description
Technical field
The invention belongs to environment and technical field of new energies, be specifically related to a kind of improve Clostridium beijerinckii produce electricity method and
Application.
Background technology
Along with the fast development of economic globalization, various countries are to energy demand expanding day, and lack of energy becomes the difficulty in the world
Topic, when facing this problem, eye is put into a cyclopean family microorganism by scientist.Microorganism is utilized to produce
Electricity, solves energy shortage problem.At present, microbiological fuel cell (Microbial Fuel Cell, MFC) technology becomes day by day
Ripe, and paid attention to.
But, the electricity generation ability of microorganism differs greatly, and produces the function of microbiological fuel cell during electricity microorganism but determines
And application.Through report, high yield electricity microorganism be still concentrated mainly on genus Shewanella (Shewanella) and ground Bacillus
(Geobacter) some gram negative bacterias such as (Energy Environ. Sci., 2011,4,4366 4379).And
The gram positive bacterias such as Clostridium beijerinckii are reported the fewest, Sophie Peguin et al. (Biotechnology and
Bioengineering, Vol. 51, Pp. 342-348 (1996)) utilize three-electrode system to have studied clostridium acetobutylicum
Electricity situation is produced as the microbiological fuel cell in the case of electron mediator at methyl viologen；Liu Jun et al.
(Biotechnology Lett, 2015,37:95-100) also utilizes 0.15g/L methyl viologen as electron mediator, 1g/L Portugal
Grape sugar is as obtaining maximum voltage 230mV, peak power output density 79.2 mW/m during carbon source2.Visible, Clostridium beijerinckii etc. is removed from office
The application in microbiological fuel cell of the Lan Shi positive bacteria causes attention day by day.
Self electricity generation ability of known microorganisms seriously hinders the development of MFC technology, but by genetic engineering modified
Thalline, it is possible to accelerate the metabolism of cell, improves electron transport rate and quantity.Based on this, genetic engineering means is to promoting micro-life
The development of thing fuel cell and commercial application will have considerable prospect.
Summary of the invention
It is an object of the invention to provide a kind of Clostridium beijerinckii that improves and produce electric method.
Another object of the present invention is to provide the Clostridium beijerinckii of a kind of high activity electricity generation ability preparing Microbial fuel
Application in battery.
The technical solution used in the present invention is:
A kind of Clostridium beijerinckii that improves produces electric method, the function of Clostridium beijerinckii PROTEIN C bei_3304 is lacked.
Further, method described above, make in Clostridium beijerinckii the gene of encoding proteins Cbei_3304 can not normal table
Reach.
Further, method described above, carry out the gene of encoding proteins Cbei_3304 in Clostridium beijerinckii inserting and lose
Live.
Further, method described above, by the base sequence of encoding proteins Cbei_3304 as shown in SEQ ID No:1.
Further, above-mentioned Clostridium beijerinckii isClostridium beijerinckii NCIMB 8052。
The application in producing electricity of a kind of high yield electricity Clostridium beijerinckii, described high yield electricity Clostridium beijerinckii is for lacking normal albumen
The Clostridium beijerinckii of Cbei_3304 function.
The application in preparing microbiological fuel cell of a kind of high yield electricity Clostridium beijerinckii, described high yield electricity Clostridium beijerinckii is scarce
Lose the Clostridium beijerinckii of normal PROTEIN C bei_3304 function.
Further, described high yield electricity Clostridium beijerinckii is the Clostridium beijerinckii that method described above prepares.
Further, above-mentioned high yield electricity Clostridium beijerinckii with glucose sugar, xylose, sucrose, fructose or starch as electron donor,
Further, above-mentioned high yield electricity Clostridium beijerinckii is with methyl viologen, methylene blue or anthraquinone-2, and 6-sodium disulfonate is as electricity
Sub-amboceptor.
The invention has the beneficial effects as follows:
The present invention inactivates by carrying out the Cbei_3304 gene encoding memebrane protein in Clostridium beijerinckii inserting so that this gene is not
Energy normal expression, thus reach to improve the purpose of Clostridium beijerinckii electricity generation ability.The invention provides one simply, improve efficiently
Clostridium beijerinckii produces the method for electricity, and the recombinant bacterial strain obtained by the method is in 1 g/L glucose seed liquor, and 0.4 g/L(is optimal
Concentration) methyl viologen (MV) is when being electron mediator, maximum output voltage is up to 409mV, compares initial strainsClostridium beijerinckiiNCIMB 8052 improves 3.13 times, and peak power output density is 144.4mW/m2, compare initial strainsClostridium beijerinckiiNCIMB 8052 improves 1.71 times, and (in 8052-MFC, optimal methyl viologen concentration is
0.1g/L).
The technique of the present invention is simple and convenient to operate, pollution-free, low cost, and its fuel utilization is general relatively wide, and electron recovery efficiency is relatively
Height, alleviates energy crisis, and high yield electricity Clostridium beijerinckii of the present invention is that a strain is suitable for microbiological fuel cell research and application
Strain excellent.
Accompanying drawing explanation
Fig. 1 is the plasmid map that the present invention inserts inactivating vectors pWJ；
Fig. 2 is the mechanism figure that the present invention uses two type Intron insertion inactivations；
Fig. 3 is transformant bacterium colony PCR electrophoretogram；
Fig. 4 is the Clostridium beijerinckii 3304 of the present invention electroactive figure of product in 1g/L glucose；
Fig. 5 is the initial strains NCIMB 8052 electroactive figure of the product in 1g/L glucose；
Fig. 6 is that high yield electricity Clostridium beijerinckii of the present invention utilizes, with initial strains NCIMB 8052, the product electricity feelings that 1g/L glucose is fuel
Condition.
Detailed description of the invention
A kind of Clostridium beijerinckii that improves produces electric method, the function of Clostridium beijerinckii PROTEIN C bei_3304 is lacked.
Preferably, method described above, make the gene of encoding proteins Cbei_3304 in Clostridium beijerinckii can not normal expression.
Preferably, method described above, carry out the gene of encoding proteins Cbei_3304 in Clostridium beijerinckii inserting inactivation.
Preferably, method described above, by the base sequence of encoding proteins Cbei_3304 as shown in SEQ ID No:1.
Preferably, above-mentioned Clostridium beijerinckii isClostridium beijerinckii NCIMB 8052。
The application in producing electricity of a kind of high yield electricity Clostridium beijerinckii, described high yield electricity Clostridium beijerinckii is for lacking normal albumen
The Clostridium beijerinckii of Cbei_3304 function.
Preferably, described high yield electricity Clostridium beijerinckii is the Clostridium beijerinckii that method described above prepares.
The application in preparing microbiological fuel cell of a kind of high yield electricity Clostridium beijerinckii, described high yield electricity Clostridium beijerinckii is scarce
Lose the Clostridium beijerinckii of normal PROTEIN C bei_3304 function.
Preferably, described high yield electricity Clostridium beijerinckii is the Clostridium beijerinckii that method described above prepares.
Preferably, above-mentioned high yield electricity Clostridium beijerinckii with glucose sugar, xylose, sucrose, fructose or starch as electron donor,
Preferably, above-mentioned high yield electricity Clostridium beijerinckii is with methyl viologen, methylene blue or anthraquinone-2, and 6-sodium disulfonate is as electronics
Amboceptor.
Preferably, above-mentioned high yield electricity Clostridium beijerinckii carries out producing electricity through dual chamber MFC.
Below in conjunction with specific embodiment, the present invention is further illustrated, but is not limited thereto.
Embodiment 1
The structure of Clostridium beijerinckii Cbei_3304 gene disruption mutant
Build the principle of Clostridium beijerinckii Cbei_3304 gene disruption mutant as in figure 2 it is shown, concrete building process include with
Lower step:
(1) Cbei_3304 inserts the structure of inactivating vectors
1) design intron
According to the Cbei_3304 gene order (as shown in SEQ ID No:1) of the Clostridium beijerinckii that ncbi database is included, by soft
Gene loci (http://www.clostron.com) is suitably inserted in part design, select to be inserted in the 101-102 base it
Between, and generate intron sequences, synthesis its sequence of intron sequences S-304(is as shown in SEQ ID NO:2), and design following drawing
Thing.
Cloning primer:
PWJ-OSC-304-S:5 '-ggagtgtcgaggatcctcgagAtaattatccttacacttcgcc-3 ', its sequence
As shown in SEQ ID NO:3；
PWJ-OSC-304-A:5 '-ggttctcctacagattgtacaAatgtggtgataacagataag-3 ', its sequence is such as
Shown in SEQ ID NO:4.
Checking primer:
Cbei-3304-T-S:5 '-taaattacctacagcaaaactgtg-3 ', sequence is as shown in SEQ ID NO:5；
Cbei-3304-T-A:5 '-ggaattaagaaccttgaatctatc-3 ', sequence is as shown in SEQ ID NO:6.
Primer pWJ-OSC-304-S introduces Xho I restriction enzyme site (underscore part), and primer pWJ-OSC-304-A introduces
BsrG I restriction enzyme site (underscore part).
2) Cbei_3304-pWJ-304 construction of recombinant vector
With Xho I and BsrG I double digestion carrier pWJ, pWJ plasmid map is as it is shown in figure 1, its sequence is as shown in SEQ ID NO:7.
The purified test kit of digestion products (Takara) after purification, passes through one-step cloning (ClonExpress) with intron sequences S-304
Connect.By the recombinant plasmid transformed of one-step cloning connection to escherichia coli E.coliDH5a, is applied to containing 50 μ g/ml ammonia benzyls
Chloramphenicol resistance LB flat board, 37 DEG C of cultivations 12～16h, picking transformant, receive liquid and contain 50 μ g/ml ammonia benzyl mycin LB cultivations
In base, 37 DEG C, 200rpm cultivate 12h, extract recombiant plasmid (AXYGEN), sequence verification, it is thus achieved that Cbei_3304-pWJ-304 weight
Group carrier.
3) the methylating of Cbei_3304-pWJ-304 recombinant vector
Preparation E.coliThe Competent of Top 10/pAN2, will check order successful Cbei_3304-pWJ-304 recombinant vector
It is transformed into escherichia coli E.coliTop 10, owing to pAN2 plasmid has tetracyclin resistance, therefore is applied to containing 50 μ g/ml ammonia
Benzyl mycin and 10 μ g/ml tetracycline Double LB flat boards, 37 DEG C of cultivations 12～16h, picking transformant, receive liquid and contain 50 μ
In g/ml ammonia benzyl mycin and 10 g/ml tetracycline LB culture medium, 37 DEG C, 200rpm cultivate 12h, extract methylated Cbei_
3304-pWJ-304 recombinant vector (pAN2 plasmid contains a bacillus subtilis phage gene, can encode transmethylase,
Exogenous plasmid methylating in escherichia coli can be realized), i.e. Cbei_3304 inserts inactivating vectors.
(2) Cbei_3304 insert inactivating vectors convert Clostridium beijerinckii (Clostridium beijerinckii NCIMB
8052)
1) willClostridium beijerinckiiNCIMB 8052 is seeded to YPS culture medium (yeast powder 3g/L, albumen
Peptone 5g/L, glucose 1g/L, ammonium acetate 2g/L, sodium chloride 3g/L, bitter salt 3g/L, potassium dihydrogen phosphate 1g/L, phosphorus
Acid hydrogen dipotassium 1g/L, green vitriol 0.1g/L, pH=6.) 37 DEG C of incubated overnight, it is inoculated into YPS training with 5% ratio next day
Support base, cultivate 6-8h, be inoculated into 2 × YTG culture medium (yeast powder 16 g/L, peptone 10 g/L, glucose 5 with 10% for 37 DEG C
G/L, sodium chloride 5 g/L) 37 DEG C cultivate 3h, OD600nm=1；
2) take 50ml Clostridium beijerinckii bacterium solution, 5000rpm, 4 DEG C of centrifugal 10 min, abandon supernatant.With ETM buffer (270mM sugarcane
Sugar, 0.6mM Na2HPO4, 4.4mM Na2HPO4, 10mM MgCl2) resuspended；Ibid it is centrifuged, removes supernatant, again with ETM buffer
Resuspended, ibid it is centrifuged, thoroughly takes supernatant；
3) with 1ml ET buffer (270mM sucrose, 0.6mM Na2HPO4, 4.4mM NaH2PO4Resuspended, take 200 l, add
1ug Cbei_3304 inserts inactivating vectors, adds the electric revolving cup of 0.2cm pre-cooling, mixes gently；
4) using MicroPulserTM electroporation electricity to turn, condition is voltage 1.8kV, resistance 200 Ω, and electric capacity 2.5 F, after electric shock
It is added immediately 1mL 2 × YTG culture medium, transfers to recovery 2～3h in sterile centrifugation tube；
5) take the 200 above-mentioned bacterium solution of l, be applied to the YPS solid medium containing 10 g/ml erythromycin, cultivate 2～3 days.
(3) Cbei_3304 inserts the screening of Inactivating mutations strain
Picking above-mentioned steps 5) the middle transformant cultivating 2～3 days, use primer Cbei-3304-T-S and Cbei-3304-T-A pair
Transformant carries out bacterium colony PCR checking, and (after insertion, PCR amplifies gene band to filter out the mutant of Intron insertion genome
Than wild type about 1Kbp on electrophoretogram), as it is shown on figure 3, the mutant being correctly inserted into is passed on three times, be coated on simultaneously containing
Erythromycinresistant and do not have on the YPS solid medium of Erythromycinresistant, filters out and knocks out the mutant of plasmid loss (red mould
The mutant that can not grow in element resistant panel), obtain high yield electricity Clostridium beijerinckii of the present invention.
Embodiment 2
This example demonstrates that the high yield electricity Clostridium beijerinckii of above-mentioned structure utilizes the product electricity of 1g/L glucose sugar in fact as anode catalyst
Test.
(1) microbiological fuel cell is built
The present embodiment establishes the microorganism combustion utilizing Clostridium beijerinckii to generate electricity for anode catalyst according to existing technology and method
Material battery, including anode chamber, cathode chamber, PEM and four parts of external circuit.Anode electrode and cathode electrode are PAN
Base graphite soft felt (5 × 5cm), using titanium silk as external circuits, outer meeting resistance is 1000 Ω, and PEM is Du Pont
NafionN117, using data acquisition unit is Keithley series.
(2) culture medium prescription:
YPS seed culture medium: yeast powder 3g/L, peptone 5g/L, glucose 1g/L, ammonium acetate 2g/L, sodium chloride 3g/L,
Bitter salt 3g/L, potassium dihydrogen phosphate 1g/L, dipotassium hydrogen phosphate 1g/L, green vitriol 0.1g/L, pH=6.
Dual chamber MFC Anodic liquid: yeast powder 3g/L, peptone 5g/L, glucose 1g/L, ammonium acetate 2g/L, sodium chloride
3g/L, bitter salt 3g/L, potassium dihydrogen phosphate 1g/L, dipotassium hydrogen phosphate 1g/L, green vitriol 0.1g/L, methyl
Purpurine (in 3304-MFC, MV concentration is 0.4g/L, and in 8052-MFC, MV concentration is 0.1g/L), pH=6.
Catholyte in dual chamber MFC: two hypophosphite monohydrate sodium dihydrogen 2.772g/L, disodium hydrogen phosphate dodecahydrate 11.5g/L, chlorine
Change potassium 0.13g//l, potassium ferricyanide 40mM, pH=7.
(3) in YPS seed liquor, activate the high yield electricity Bai Shi shuttle that high yield electricity recombinant bacterial strain 3304(embodiment 1 builds respectively
Bacterium) and initial strains 8052(Clostridium beijerinckiiNCIMB 8052), cultivation temperature 37 DEG C, Anaerobic culturel
Time 12h.Then carry out 250mL dual chamber MFC technology, in anode chamber, access anolyte 225mL, methyl viologen (MV), be passed through
N2 3 minutes, then access the Clostridium beijerinckii (inoculum concentration 10%(v/v) of 25mL), it is passed through N2 3 minutes, discharge the oxygen in solution, rapidly
Seal anode chamber, add catholyte to cathode chamber, seal rapidly cathode chamber, cultivation temperature 30 DEG C, produce electro-detection time 33h,
Obtain result eventually as shown in 1 table and Fig. 4 ~ 6.
Table 1 high yield of the present invention electricity Clostridium beijerinckii and the testing result of starting strain electricity generation performance
According to output voltage data, calculate electron recovery efficiency and obtain the polarization curve producing electricity, experimental result such as table 1 and Fig. 4 ~
Shown in 6；Wherein, Fig. 4 is the high yield electricity Clostridium beijerinckii of the present invention electroactive figure of product in 1g/L glucose；Fig. 5 is initial strains
The electroactive figure of the NCIMB 8052 product in 1g/L glucose；Fig. 6 is high yield electricity Clostridium beijerinckii of the present invention and initial strains NCIMB
8052 utilize the product electricity situation that 1g/L glucose is fuel.It can be seen that high yield electricity Clostridium beijerinckii maximum of the present invention output electricity
Pressure up to 409mV, improves 3.13 times than initial strains NCIMB 8052, and peak power output density is 144.4mW/m2, than
Initial strains NCIMB 8052 improves 1.71 times.
The present invention is in research process, it was found that glucose sugar replaces with xylose, sucrose, fructose or starch and supplies as electronics
Body, high yield electricity Clostridium beijerinckii of the present invention the most also can produce the maximum output voltage height of similar effect, peak power output density；
Methyl viologen replaces with methylene blue, anthraquinone-2, and 6-sodium disulfonate is as electron mediator, and high yield electricity Clostridium beijerinckii of the present invention is same
Sample also can play the electricity generation performance of similar effect.
The above results explanation the invention provides a kind of method improving Clostridium beijerinckii product electricity simple, efficient, obtained
High yield electricity Clostridium beijerinckii can be used for produce electricity, in particular in the product electricity of microbiological fuel cell；Therefore can be by high yield of the present invention
Electricity Clostridium beijerinckii is for the preparation of microbiological fuel cell.
Above-described embodiment is the present invention preferably embodiment, but embodiments of the present invention are not by above-described embodiment
Limit, the change made under other any spirit without departing from the present invention and principle, modify, substitute, combine, simplify,
All should be the substitute mode of equivalence, within being included in protection scope of the present invention.
<110>Guangzhou Inst of Cane Sugar
<120>a kind of Clostridium beijerinckii that improves produces method and the application thereof of electricity
<130>
<160> 6
<170> PatentIn version 3.5
<210> 1
<211> 723
<212> DNA
<213>artificial sequence
<400> 1
atgacaaagg ttaataaatt acctacagca aaactgtgga atccgaaatc atttattata 60
ttttccgtat ttttctcatt tctgccagct ggaatcatgt gtgctttgaa ttatggacga 120
tctggaagtc aaaaaaagaa gtggatattt cttttagcaa gcatcttagt gtttatagca 180
cttatagcac ttctgcctat attatcaatt aatacttcta tcatattttt cagtattaat 240
atagctttag gcataattct tatgtttaca cagttaaagc tgtataatga acatattcaa 300
aatggcgggc aaagtgcttc gtatttattt cctataatca tagggctttt gattttttca 360
ttgtcagctg cttcaattct atactcaatt tacgtgccaa aaaatgctct tgattatggc 420
gaaaatcact tattttatac taataagata acagaatccc aagcgaaaaa acttggagat 480
tatttaaatt cagaaggata cttcactcct agttcaaaag tggacgttaa gattgataaa 540
caagatacac tctatatttt atctctcgtt gtagaaggtg actataaaag tgatacaagc 600
tatgtagaac ctatgaaagc aatatcaaaa gagatttcaa aaaatgtttt tgaaaataat 660
aaagttagaa tagatttatg caatgataga ttcaaggttc ttaattccat caatgttgat 720
taa 723
<210> 2
<211> 344
<212> DNA
<213>artificial sequence
<400> 2
ctcgagataa ttatccttac acttcgcccg cgtgcgccca gatagggtgt taagtcaagt 60
agtttaaggt actactctgt aagataacac agaaaacagc caacctaacc gaaaagcgaa 120
agctgatacg ggaacagagc acggttggaa agcgatgagt tacctaaaga caatcgggta 180
cgactgagtc gcaatgttaa tcagatataa ggtataagtt gtgtttactg aacgcaagtt 240
tctaatttcg gttaagtgtc gatagaggaa agtgtctgaa acctctagta caaagaaagg 300
taagttatgg cgggcgactt atctgttatc accacatttg taca 344
<210> 3
<211> 43
<212> DNA
<213>artificial sequence
<400> 3
ggagtgtcga ggatcctcga gataattatc cttacacttc gcc 43
<210> 4
<211> 42
<212> DNA
<213>artificial sequence
<400> 4
ggttctccta cagattgtac aaatgtggtg ataacagata ag 42
<210> 5
<211> 24
<212> DNA
<213>artificial sequence
<400> 5
taaattacct acagcaaaac tgtg 24
<210> 6
<211> 24
<212> DNA
<213>artificial sequence
<400> 6
ggaattaaga accttgaatc tatc 24
Claims (10)
1. one kind is improved the method that Clostridium beijerinckii produces electricity, it is characterised in that the function of Clostridium beijerinckii PROTEIN C bei_3304 carried out
Disappearance.
Method the most according to claim 1, it is characterised in that make the gene of encoding proteins Cbei_3304 in Clostridium beijerinckii
Can not normal expression.
Method the most according to claim 2, it is characterised in that by the gene of encoding proteins Cbei_3304 in Clostridium beijerinckii
Carry out inserting inactivation.
Method the most according to claim 1, it is characterised in that by the base sequence such as SEQ of encoding proteins Cbei_3304
Shown in ID No:1.
Method the most according to claim 1, it is characterised in that described Clostridium beijerinckii isClostridium beijerinckii NCIMB 8052。
6. a high yield electricity Clostridium beijerinckii application in producing electricity, described high yield electricity Clostridium beijerinckii is for lacking normal PROTEIN C bei_
The Clostridium beijerinckii of 3304 functions.
7. a high yield electricity Clostridium beijerinckii application in preparing microbiological fuel cell, described high yield electricity Clostridium beijerinckii is disappearance
The Clostridium beijerinckii of normal PROTEIN C bei_3304 function.
Application the most according to claim 7, it is characterised in that described high yield electricity Clostridium beijerinckii is that claim 1 ~ 5 is arbitrary
The Clostridium beijerinckii that described method prepares.
Application the most according to claim 7, it is characterised in that described high yield electricity Clostridium beijerinckii is with glucose sugar, xylose, sugarcane
Sugar, fructose or starch are electron donor.
Application the most according to claim 7, it is characterised in that described high yield electricity Clostridium beijerinckii is with methyl viologen, methylene
Indigo plant or anthraquinone 2,6 disulfonic acid sodium are as electron mediator.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610399488.9A CN106047754B (en) | 2016-06-07 | 2016-06-07 | A kind of method and its application improving Clostridium beijerinckii electricity production |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610399488.9A CN106047754B (en) | 2016-06-07 | 2016-06-07 | A kind of method and its application improving Clostridium beijerinckii electricity production |
Publications (2)
| Publication Number | Publication Date |
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| CN106047754A true CN106047754A (en) | 2016-10-26 |
| CN106047754B CN106047754B (en) | 2019-07-19 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107164292A (en) * | 2017-06-26 | 2017-09-15 | 南京中泰生物科技有限公司 | One plant of Clostridium beijerinckii and its construction method and application to the efficient dechlorination of pentachlorophenol |
| CN107287145A (en) * | 2017-07-31 | 2017-10-24 | 南京中泰生物科技有限公司 | One plant of electricity production Clostridium beijerinckii and its construction method and application |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103898031A (en) * | 2014-04-21 | 2014-07-02 | 南京工业大学 | Clostridium beijerinckii with high electricity yield and application thereof |
| CN103911334A (en) * | 2014-04-16 | 2014-07-09 | 广州甘蔗糖业研究所 | Clostridium beijerinckii with high stress resistance and application thereof |
-
2016
- 2016-06-07 CN CN201610399488.9A patent/CN106047754B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103911334A (en) * | 2014-04-16 | 2014-07-09 | 广州甘蔗糖业研究所 | Clostridium beijerinckii with high stress resistance and application thereof |
| CN103898031A (en) * | 2014-04-21 | 2014-07-02 | 南京工业大学 | Clostridium beijerinckii with high electricity yield and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| GENBANK: "Clostridium beijerinckii NCIMB 8052，complete genome;Genbank:CP000721.1", 《NCBI》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107164292A (en) * | 2017-06-26 | 2017-09-15 | 南京中泰生物科技有限公司 | One plant of Clostridium beijerinckii and its construction method and application to the efficient dechlorination of pentachlorophenol |
| CN107164292B (en) * | 2017-06-26 | 2020-10-16 | 南京中环生物科技有限公司 | Clostridium beijerinckii for efficiently dechlorinating pentachlorophenol and construction method and application thereof |
| CN107287145A (en) * | 2017-07-31 | 2017-10-24 | 南京中泰生物科技有限公司 | One plant of electricity production Clostridium beijerinckii and its construction method and application |
| CN107287145B (en) * | 2017-07-31 | 2020-05-01 | 南京中泰生物科技有限公司 | Clostridium beijerinckii for generating electricity and construction method and application thereof |
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| CN106047754B (en) | 2019-07-19 |
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