CN102586138A - Method for improving carbon sequestration efficiency of non-photosynthesizing microorganism by using mixed electron donor - Google Patents

Method for improving carbon sequestration efficiency of non-photosynthesizing microorganism by using mixed electron donor Download PDF

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CN102586138A
CN102586138A CN2012100128326A CN201210012832A CN102586138A CN 102586138 A CN102586138 A CN 102586138A CN 2012100128326 A CN2012100128326 A CN 2012100128326A CN 201210012832 A CN201210012832 A CN 201210012832A CN 102586138 A CN102586138 A CN 102586138A
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electron donor
microorganism
trace element
substratum
described step
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胡佳俊
王磊
张士萍
席雪飞
单伊娜
胡煜
张艳楠
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Tongji University
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Tongji University
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Abstract

The invention relates to the field of CO2 sequestration of microorganism, in particular relates to a method for improving carbon sequestration efficiency of non-photosynthesizing microorganism. The method comprises the following steps of: (1) preparing a culture medium for culturing autotrophic microorganisms, and adding a trace element solution to the culture medium; (2) preparing an electron donor concentrated solution; (3) adding the electron donor concentrated solution prepared in the step (2) into the culture solution containing trace element prepared in the step (1); and (4) adding mixed carbon sequestration strain from the ocean into the culture medium obtained in the step (3), and culturing in an aerobic condition. The method provided by the invention also has the advantages of simple technology and strong operability, and brings certain economic benefits.

Description

A kind of method of utilizing the mixed electronic donor to improve the solid carbon efficiencies of non-photosynthesizing microorganism
Technical field
The present invention relates to mikrobe fixation of C O 2The field relates to a kind of method that improves the solid carbon efficiencies of non-photosynthesizing microorganism.
Background technology
CO 2The climate warming that " Greenhouse effect " that cause are caused is the great environmental problem that the current whole world faces.Point out according to IPCC the 4th assessment report: at a nearest decade (1995～2004 years), CO 2(the annual 9.2 hundred million tons of CO that advance the speed of equivalent discharging 2Equivalent) than the drainage rate of previous decade (1970～1994 years) (annual 4.3 hundred million tons of CO 2Equivalent) much higher." the 12 planning proposal draft " of China shows that China is with CO 2Reduction of discharging has been placed on the considerable position.And on the Cancun meeting of holding for the year ends 2010, China promises to undertake the year two thousand twenty, unit gross domestic product (GDP) CO 2Discharging will be than decline 40%-45% in 2005.While CO 2Be again carbon resource the abundantest on the earth, can change it into resource and the energy.Therefore, CO 2Be fixed on environment, the energy, resource aspect all have great importance.Current, considering how to reduce discharging CO 2Prerequisite under, research CO 2Recovery with fixing, can effectively reduce free CO in the environment 2, can it be regenerated as resource again, therefore caused the extensive interest of countries in the world.
CO 2Fixedly mainly contain physics forensic chemistry method and biological process, and most of physics method and chemical method all must connect and biological process comes final fixation of C O 2Biological process fixation of C O 2Mainly be to rely on plant and autotrophic microorganism, the photosynthesis of plants outbalance is also more paid attention to by the people traditionally.But there are various environment on the earth; Particular surroundings and the occasion (like the capture occasion of arid barren desert soil and industrial gaseous waste) that can not grow plant; The advantage of the environmental compatibility that Institute of Micro-biology has has just displayed; Therefore substance flow and the energy from whole biosphere flows mikrobe fixation of C O 2Significant.
If generally acknowledge at present and have higher microbial host photosynthetic microorganism of carbon efficiencies admittedly and the hydrogen-oxygen bacterium in the chemosynthetic bacteria.Photosynthetic microorganisms such as algae need illumination because in culturing process, with and thermo-labile and high concentration CO 2Characteristic, limited its practical application.Though and the hydrogen-oxygen bacterium is comparatively wide in range without illumination and growth scope; But it must provide with high concentration hydrogen in process of growth as electron donor; Therefore the conventional environment condition is difficult to meet its growth requirement, and in practical application, also there is serious potential safety hazard in hydrogen supply gas simultaneously.Given this, excavate efficient carbon mikrobe admittedly, for realizing that the solid carbon of mikrobe under the conventional environment condition is (like edatope and absorption industrial discharge CO without illumination and hydrogen supply 2The macro-organism reactor drum in) significant.
There is critical role the ocean in the global carbon process, will absorb 2.0Gt (1Gt=10 its every year 9T) anthropogenic discharge's CO 2, the research of solid carbon mikrobe also is the focus that global association area expert pays close attention to always in the ocean.For this reason, we gather the water and soil sample in a plurality of marine sites (comprising global four ocean, the dozens of countries and regions) from the whole world, have obtained the solid carbon microorganism species without illumination and hydrogen supply of a plurality of series through separation screening.It is lower that but the solid carbon efficiencies of these floras is compared photosynthetic microorganism; Its major cause is that the available energy substance kind of these floras is limited, the energy substance amount is limited and utilizability energy substance is limited; If utilize mixed electronic donor system then can effectively address these problems; Its solid carbon efficiencies be will help significantly improving, flora potential economic benefit and social benefit in practical application further strengthened.
Summary of the invention
The objective of the invention is to provides the method for utilizing the mixed electronic donor to improve the solid carbon efficiencies of non-photosynthesizing microorganism under a kind of aerobic condition for the defective that overcomes prior art.
For realizing above-mentioned purpose, the technical scheme that the present invention adopted is:
A kind of method that improves the solid carbon efficiencies of non-photosynthesizing microorganism comprises following steps:
(1) substratum of autotrophic microorganism is cultivated in preparation; In above-mentioned substratum, add trace element solution then again;
(2) preparation electron donor liquid concentrator;
(3) the electron donor liquid concentrator that makes in the step (2) is joined in the nutrient solution that contains trace element that makes in the step (1);
(4) obtain adding in the substratum from the solid carbon bacterial classification of the mixing of ocean to step (3), under aerobic condition, cultivate.
Substratum in the described step (1) comprises following component: 0.5-1.0g/L KH 2PO 4, 1.0-2.0g/L K 2HPO 4, 0.1-0.2g/L MgSO 47H 2O, 10-30g/L NaCl, 0.001-0.01g/L CaCl 2, 0.0036mmol-0.036mmol/L ferrous salt and 0.00375-0.0375mol/L NH 4 +
Described ferrous salt is selected from FeSO 47H 2O, FeSO 4Or FeCl 2
Described ammonium salt is selected from (NH 4) 2SO 4Or NH 4Cl.
Described trace element solution comprises 1.68mg/LNa 2MoO 42H 2O, 0.4mg/LH 3BO 3, 1.0mg/LZnSO 47H 2O, 1.0mg/LMnSO 45H 2O, 7.0mg/LCuSO 45H 2O, 1.0mg/LCoCl 26H 2O or 1.0mg/LNiSO 47H 2Among the O more than one.
In the described step (1), add the 2ml trace element solution in every liter of nutrient solution.
Electron donor is selected from MnSO in the described step (2) 45H 2In O, nitrite, thiosulphate or the sulfide one or more.
Described nitrite is selected from NaNO 2Or KNO 2
Described thiosulphate is selected from Na 2S 2O 3Or K 2S 2O 3
Described sulfide is selected from Na 2S or K 2S.
In the described step (2), the concentration of every kind of electron donor is 80-200mg/mL in the electron donor liquid concentrator.
The consumption of mixed electronic donor is 0.01-10g/L MnSO in the described step (3) 45H 2O, 1-15g/L nitrite, 1-15g/L thiosulphate or 1-15g/L sulfide.
Be selected from obligate/facultative autotrophy mikrobe and heterotrophic microorganism from the solid carbon bacterial classification of the mixing of ocean in the described step (4); Be selected from the sedimental mixing microorganisms flora of ocean seawater or seawater; This flora mainly is made up of the chemoautotrophy mikrobe, includes in iron bacteria, hydrogen bacterium, thiobacterium, manganese bacteria or the nitrobacteria more than one.
The aerobic condition of described step (4) is: oxygen content is 5-25% in the gas, and carbon dioxide content is greater than 0%, the preferred 5-30% of carbon dioxide content, and further the content of preferably carbon dioxide is 20%, incubation time is 4-8 days.
Beneficial effect of the present invention is:
With MnSO 45H 2O, NaNO 2, KNO 2, Na 2S 2O 3Or Na 2Among the S more than one are processed mixed electronic donor system, can effectively promote the solid carbon efficiencies of non-photosynthetic carbon fixation mikrobe; Show through experiment, use the mixed electronic donor to cultivate non-photosynthetic carbon fixation mikrobe, the several times that its solid carbon efficiencies is to use single electron donor to cultivate are as with NaNO 2, Na 2S 2O 3And Na 2The effect of the mixed electronic donor system that S forms is not add MnSO 45H 2O, NaNO 2, KNO 2, K 2S 2O 3, Na 2S 2O 3, K 2S or Na 2Among the S 6424% of any effect, show that the mixed electronic donor can be effectively carries out synergy to the solid carbon efficiencies of mikrobe, thereby realize CO 2Resource utilization.It is simple, workable and have an advantage of certain economic benefit that the present invention also has technology.
Embodiment
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used to the present invention is described and be not used in the restriction scope of the present invention.Should be understood that in addition those skilled in the art can do various changes or modification to the present invention after the content of having read the present invention's instruction, these equivalent form of values fall within the application's appended claims institute restricted portion equally.
Embodiment 1
(1) substratum of autotrophic microorganism is cultivated in preparation
Culture medium prescription is (g/L) as follows: KH 2PO 4(1.0); K 2HPO 4(2.0); MgSO 47H 2O (0.2); NaCl (20) and CaCl 2(0.01); (NH 4) 2SO 4(0.038mol/L); FeSO 47H 2O (0.0036mmol/L); Go up whenever and to state the trace element solution that adds 2mL in the substratum again.
Trace element solution is selected from and comprises Na 2MoO 42H 2O (1.68); H 3BO 3(0.4); ZnSO 47H 2O (1.0); MnSO 45H 2O (1.0); CuSO 45H 2O (7.0); CoCl 26H 2O (1.0) and NiSO 47H 2O (1.0), concentration is in mg/L;
(2) respectively separately after the preparation as the NaNO of electron donor 2, Na 2S 2O 3And Na 2The liquid concentrator of S, its concentration is respectively 80mg/mL;
(3) choose in the electron donor liquid concentrator that step (2) obtains several kinds, according to 4.6g/L NaNO 2, 5.0g/L Na 2S 2O 3With 12.5g/L Na 2The consumption of S adds the electron donor liquid concentrator that step (2) obtains in the substratum that step (1) obtains;
(4) will screen from a plurality of oceans seawater sample and be inoculated in step (3) and obtain in the substratum through the solid carbon microorganism species of the acclimation shaking culture of half a year; And under aerobic condition, cultivated 4 days; Wherein, Wherein mixed gas is made up of air and carbonic acid gas, its air: carbonic acid gas=80: 20, volume ratio.Solid carbon microorganism species in the present embodiment is specifically: screening is seawater or the sedimental mixing microorganisms flora of seawater from the ocean, and this flora mainly is made up of the chemoautotrophy mikrobe, mainly includes thiobacterium, nitrobacteria etc.
Embodiment 2
(1) substratum of autotrophic microorganism is cultivated in preparation.Culture medium prescription is (g/L) as follows: KH 2PO 4(1.0); K 2HPO 4(2.0); MgSO 47H 2O (0.2); NaCl (20) and CaCl 2(0.01); (NH 4) 2SO 4(0.038mol/L); FeSO 47H 2O (0.0036mmol/L) goes up whenever and states the trace element solution that adds 2mL in the substratum again.
Trace element solution is selected from and comprises Na 2MoO 42H 2O (1.68); H 3BO 3(0.4); ZnSO 47H 2O (1.0); MnSO 45H 2O (1.0); CuSO 45H 2O (7.0); CoCl 26H 2O (1.0) and NiSO 47H 2O (1.0), concentration is in mg/L;
(2) respectively separately after the preparation as the NaNO of electron donor 2, Na 2S 2O 3And Na 2The liquid concentrator of S, its concentration is respectively 160mg/mL;
(3) choose in the electron donor liquid concentrator that step (2) obtains several kinds, according to 0.01g/L MnSO 45H 2O, 5.0g/LNaNO 2, 5.0g/L Na 2S 2O 3With 12.5g/L Na 2The consumption of S adds the electron donor liquid concentrator that step (2) obtains in the substratum that step (1) obtains.
(4) a plurality of solid carbon microorganism species that will screen from Chinese Xiamen, Chinese Hainan, Chinese Shanghai, Chinese Qingdao, Australia, Phuket, THA, Japanese celestial platform, France add Lay, Papua New Guinea, the South Pole and Arctic Sea water sample are inoculated in step (3) and obtain in the substratum; And under aerobic condition, cultivated 4 days; Wherein mixed gas is made up of air and carbonic acid gas; Its air: carbonic acid gas=80: 20, volume ratio.
Bacteria screening in the present embodiment is seawater or the sedimental mixing microorganisms flora of seawater from the ocean, and this flora mainly is made up of the chemoautotrophy mikrobe, and the chemoautotrophy mikrobe includes hydrogen bacterium, thiobacterium, nitrobacteria.
Embodiment 3
(1) substratum of autotrophic microorganism is cultivated in preparation.Culture medium prescription is (g/L) as follows: KH 2PO 4(1.0); K 2HPO 4(2.0); MgSO 47H 2O (0.2); NaCl (20) and CaCl 2(0.01); NH 4Cl (0.019mol/L); FeCl 2(0.00144mmol/L); Go up whenever and to state the trace element solution that adds 2mL in the substratum again.Trace element solution is selected from and comprises Na 2MoO 42H 2O (1.68); H 3BO 3(0.4); ZnSO 47H 2O (1.0); MnSO 45H 2O (1.0); CuSO 45H 2O (7.0); CoCl 26H 2O (1.0); AndNiSO 47H 2O (1.0), concentration is in mg/L.
(2) respectively separately after the preparation as electron donor, NaNO 2, Na 2S 2O 3And Na 2The liquid concentrator of S, its concentration are 200mg/mL;
(3) choose in the electron donor liquid concentrator that step (2) obtains several kinds, according to 5.0g/L NaNO 2Perhaps 2.3g/LNa 2S 2O 3Perhaps 2g/L Na 2The electron donor consumption of any one among the S adds the electron donor liquid concentrator that step (2) obtains in the substratum that step (1) obtains;
(4) will screen from a plurality of oceans seawater sample and be inoculated in step (3) and obtain in the substratum through the solid carbon microorganism species of acclimation shaking culture of many time of half a year; And under aerobic condition, cultivated 4 days; Wherein aerobic condition specifically: be made up of air and carbonic acid gas in the mixed gas; Its air: carbonic acid gas=80: 20, volume ratio.
Bacteria screening in the present embodiment is seawater or the sedimental mixing microorganisms flora of seawater from the ocean, and this flora mainly is made up of the chemoautotrophy mikrobe, mainly includes thiobacterium, nitrobacteria etc.
Among the embodiment, for the synergistic effect of research mixed electronic donor to mikrobe, get the sample of cultivating 4 days, survey total organic carbon concentration in its nutrient solution, because initial medium total organic carbon concentration is 0, and the utilizable carbon source of mikrobe has only CO in culturing process 2This inorganic carbon source is so the total organic carbon amount that has increased in the substratum all is come from mikrobe with CO 2Fixing gained.The result of instance 1 shows, through its CO at 4 days internal fixing of mixed electronic donor cultured microorganism 2Amount has reached 387.51mg/L, and does not use fixation of C O under the same culture condition of mixed electronic donor cultured microorganism 2Efficient is merely 5.94mg/L, the former than the latter high 6424%.Experiment shows, process method of the present invention, mikrobe fixation of C O 2Efficient significantly increases.Among the result of instance 2, with the optimized electronic donor H that generally acknowledges 2Be contrast, with the cultivation of mixed electronic donor after 4 days, from the different microorganisms flora in more than ten marine sites, its fixation of C O 2Efficient is to use H 2During cultivation 380%.Among the result of instance 3, according to 5.0g/L NaNO 2Perhaps 2.3g/L Na 2S 2O 3Perhaps 2g/L Na 2The electron donor consumption cultured microorganism of any one among the S is than the fixation of C O of the direct cultured microorganism that does not add them 2Efficient will exceed one times.Experiment shows, process method of the present invention, mikrobe fixation of C O 2Efficient significantly increases.
To sum up visible, this method technology is simple, workable, and for all effective from the microorganism species in more than ten marine site, global 4 ocean, explains that this method is for microorganism species fixation of C O 2Synergistic effect have ubiquity, can effectively be used in mikrobe fixation of C O 2Process in, thereby realize CO 2Resource utilization.
The above-mentioned description to embodiment is can understand and use the present invention for ease of the those of ordinary skill of this technical field.The personnel of skilled obviously can easily make various modifications to these embodiment, and needn't pass through performing creative labour being applied in the General Principle of this explanation among other embodiment.Therefore, the invention is not restricted to the embodiment here, those skilled in the art are according to announcement of the present invention, and not breaking away from the improvement that category of the present invention makes and revise all should be within protection scope of the present invention.

Claims (10)

1. one kind is improved the non-photosynthesizing microorganism method of carbon efficiencies admittedly, it is characterized in that: comprise following steps:
(1) substratum of autotrophic microorganism is cultivated in preparation; In above-mentioned substratum, add trace element solution then again;
(2) preparation electron donor liquid concentrator;
(3) the electron donor liquid concentrator that makes in the step (2) is joined in the nutrient solution that contains trace element that makes in the step (1);
(4) obtain adding in the substratum from the solid carbon bacterial classification of the mixing of ocean to step (3), under aerobic condition, cultivate.
2. method according to claim 1 is characterized in that: the substratum in the described step (1) comprises following component: 0.5-1.0g/LKH 2PO 4, 1.0-2.0g/L K 2HPO 4, 0.1-0.2g/L MgSO 47H 2O, 10-30g/LNaCl, 0.001-0.01g/L CaCl 2, 0.0036mmol-0.036mmol/L ferrous salt and 0.00375-0.0375mol/L NH 4 +
3. method according to claim 1 is characterized in that: described ferrous salt is selected from FeSO 47H 2O, FeSO 4Or FeCl 2
Or described ammonium salt is selected from (NH 4) 2SO 4Or NH 4Cl.
4. method according to claim 1 is characterized in that: described trace element solution comprises 1.68mg/LNa 2MoO 42H 2O, 0.4mg/LH 3BO 3, 1.0mg/LZnSO 47H 2O, 1.0mg/LMnSO 45H 2O, 7.0mg/LCuSO 45H 2O, 1.0mg/LCoCl 26H 2O or 1.0mg/LNiSO 47H 2Among the O more than one.
5. method according to claim 1 is characterized in that: in the described step (1), add the 2ml trace element solution in every liter of nutrient solution.
6. method according to claim 1 is characterized in that: electron donor is selected from MnSO in the described step (2) 45H 2In O, nitrite, thiosulphate or the sulfide one or more.
7. method according to claim 6 is characterized in that: described nitrite is selected from NaNO 2Or KNO 2
Described thiosulphate is selected from Na 2S 2O 3Or K 2S 2O 3
Described sulfide is selected from Na 2S or K 2S.
8. method according to claim 1 is characterized in that: in the described step (2), the concentration of every kind of electron donor is 80-200mg/mL in the electron donor liquid concentrator;
Or the consumption of mixed electronic donor is 0.01-10g/L MnSO in the described step (3) 45H 2O, 1-15g/L nitrite, 1-15g/L thiosulphate or 1-15g/L sulfide.
9. method according to claim 1; It is characterized in that: be selected from obligate/facultative autotrophy mikrobe and heterotrophic microorganism from the solid carbon bacterial classification of the mixing of ocean in the described step (4); Be selected from the sedimental mixing microorganisms flora of ocean seawater or seawater; This flora is made up of the chemoautotrophy mikrobe, comprising in iron bacteria, hydrogen bacterium, thiobacterium, manganese bacteria or the nitrobacteria more than one are arranged.
10. method according to claim 1; It is characterized in that: the aerobic condition of described step (4) is: oxygen content is 5-25% in the gas; Carbon dioxide content is greater than 0%; The preferred 5-30% of carbon dioxide content, further the content of preferably carbon dioxide is 20%, incubation time is 4-8 days.
CN2012100128326A 2012-01-16 2012-01-16 Method for improving carbon sequestration efficiency of non-photosynthesizing microorganism by using mixed electron donor Pending CN102586138A (en)

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Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
JIA-JUN HU,ET AL: "Enhanced CO2 fixation by a non-photosynthetic microbial community under anaerobic conditions: Optimization of electron donors", 《BIORESOURCE TECHNOLOGY》 *
JIA-JUN HU,ET AL: "Optimization of electron donors to improve CO2 fixation efficiency by a non-photosynthetic microbial community under aerobic condition using statistical experimental design", 《BIORESOURCE TECHNOLOGY》 *
武满满等: "混合电子供体对好氧非光合微生物菌群固碳效率影响的析因实验分析", 《环境科学学报》 *
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Application publication date: 20120718