CN105039423A - Method for improving concentration of alcohol substances in fermented product of anaerobic gas-feeding microbes - Google Patents
Method for improving concentration of alcohol substances in fermented product of anaerobic gas-feeding microbes Download PDFInfo
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- CN105039423A CN105039423A CN201510533004.0A CN201510533004A CN105039423A CN 105039423 A CN105039423 A CN 105039423A CN 201510533004 A CN201510533004 A CN 201510533004A CN 105039423 A CN105039423 A CN 105039423A
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Abstract
The inventions aims to provide a cultivation system for improving concentration of alcohol substances in a fermented product of anaerobic gas-feeding microbes, specifically for improving the concentration of the alcohol substances in the fermented product of the anaerobic gas-feeding microbes in a shaking cultivation or continuous air agitation cultivation system by optimizing components in a cultivation medium. A method for improving the fermentation efficiency of the anaerobic gas-feeding microbes is characterized in that the final concentration of molybdate ions in the cultivation medium for fermentation is not higher than 0.1mg/L. By the method, the concentration of alcohols in the fermented product of gas biotransformation can be improved and the value of the bio-fermentation production of gas can be improved effectively.
Description
Technical field
The invention belongs to microbial gas fermentation technical field, be specifically related to a kind of culture system improving alcohols material concentration in anaerobism food gas microbial fermentation product.
Technical background
Along with China's rapid development of economy, a large amount of industrial tail gas has discharged in air in power plant, Steel Plant and other industrial and mining enterprises, not only causes serious environmental pollution, has caused series of environmental problems, also also result in serious harm to human health.CO, CO is rich in this kind of industrial tail gas
2and H
2, be the cheap raw material that a class has the manufacture petrochemical industry class chemical of Development volue.How fully to develop these industrial tail gas and not only can alleviate environmental stress, the substitute products of fossil resources can also be formed, alleviate the demand that China is growing to petroleum resources.At present mainly condensed combustion mode is passed through to the treatment process of industrial tail gas, also have part to be processed by chemical absorption catalysis method.Chemical absorption catalysis method needs the water vapour removed in industrial tail gas, needs again to reclaim organic solvent, and process is very complicated, and there is the secondary pollution to environment.Therefore, it is very necessary for probing into a new industrial tail gas application technology as the second resource.And utilize microorganism to be fixed by the carbonaceous component in industrial tail gas, the method forming desirable metabolites not only can reduce exhaust emissions and secondary pollution, product comprises the important compound such as ethanol, butanols, succinic acid, can substitute as the excellent of petroleum chemicals.Small molecular organic acid and alcohols material (formula 1-4) is comprised in the product of gas fermentable.
6CO+3H
2O＝C
2H
5OH+4CO
2(1)
6H
2+2CO
2＝C
2H
5OH+3H
2O(2)
4CO+2H
2O＝CH
3COOH+2CO
2(3)
2CO
2+4H
2＝CH
3COOH+2H
2O(4)
Alcohols wherein such as ethanol, butanols are excellent clean fuels, and 2,3 butyleneglycols and n-hexyl alcohol are also important Chemicals.Compare organic acid, have higher value, therefore people to tend to by biology more by gas reforming is alcohols material instead of organic acid.Acid alcohol in product is than being the important indicator evaluating gaseous bio fermentation, and lower acid alcohol, than the overall value that can not only improve tunning, can reduce the cost of subsequent products separation and purification simultaneously.
Summary of the invention
The object of this invention is to provide a kind of culture system improving alcohols material concentration in anaerobism food gas microbial fermentation product, namely by being optimized the component in substratum, in shaking culture or continuous aeration-agitation culture system, realize the raising of alcohols material concentration in microbial fermentation product.
The method of raising anaerobism of the present invention food gas fermentable efficiency is that the final concentration of molybdenum acid ion in the substratum being used in fermentation is not more than 0.1mg/L;
As preferably, be do not add molybdenum acid ion in the substratum for fermenting;
Further, described anaerobism food gas microorganism is preferably anaerobism food gas clostridium;
Preferred as embodiment, the substratum used includes the mineralsolutionstock (V/V) of 2%-4%, the yeast extract (M/V) of 0.05%, the morpholino b acid (M/V) of 0.5%, the reducingagentStock (V/V) of 0.5%, the micro-storage liquid (V/V) of 0.1%, selectivity adds Na
2moO
42H
2o storage liquid.
Wherein the consisting of of Mineralsolutionstock (M/V): 8% sodium-chlor, 10% ammonium chloride, 1% Repone K, the potassium primary phosphate of 1%, the magnesium sulfate of 2% and the calcium chloride of 0.4%.
ReducingAgentStock (M/V) consist of 0.9% sodium hydroxide, the L-cysteine hydrochloride of 4% and the nine water cure sodium of 4%.
Liquid microelement storage liquid (M/V) consist of 1% nitrilotriacetic acid(NTA), the magnesium sulfate of 0.5%, the ferrous ammonium sulphate of 0.4%, the cobalt chloride of 0.1%, the zinc sulfate of 0.1%, the nickelous chloride of 0.01%, the sodium selenate of 0.01% and the sodium wolframate of 0.01% and 0.02% copper sulfate.
Na
2moO
42H
2o storage liquid (M/V) to be concentration be 0.01% sodium molybdate solution.
The present invention can improve the concentration of alcohols material in gaseous bio transformation fermentation product, effectively improves the value of gaseous bio tunning.
Accompanying drawing explanation
Fig. 1: the strain growth graphic representation of embodiment 1,
Fig. 2: each Fermentation Substance Concentration figure in embodiment 1,
Fig. 3: the strain growth graphic representation of embodiment 2,
Fig. 4: the strain growth graphic representation of embodiment 3,
Fig. 5: the strain growth graphic representation of embodiment 4,
Fig. 6: each Fermentation Substance Concentration figure in embodiment 4.
Embodiment
Trace element is the important composition composition maintaining anaerobion growth metabolism and anaerobically fermenting enzyme system activity.Some trace element can promote the growth of bacterium and the activity of activating enzyme, and then promotes biosynthesizing.The stimulation of trace element and the steady running of antagonistic action to fermentation all play an important role.Applicant finds the concentration adjusting molybdenum acid ion in the substratum of anaerobism food gas clostridium in long-term research, can significantly improve the concentration of alcohols material in gaseous bio transformation fermentation product, effectively improve the value of gaseous bio tunning.
The microorganism used in the specific embodiment of the invention is anaerobism food gas Microbial biomass C lostridiumcarboxidivorans bacterial strain P7 (DSM15243), buys the German microorganism from the Leibnitz institute of Braunschweig, Germany and Cell Culture Collection.
Invent substratum used, include but not limited to the substratum of Clostridiumcarboxidivorans bacterial strain.
Below in conjunction with specific embodiment, the present invention is described in detail.
The structure explanation of basic culture system
Constant-temperature shaking culture: containing the mineralsolutionstock of 15ml in 500ml substratum, the yeast extract of 0.5g and the morpholino b acid of 2.5g, 2.5mlreducingagentStock.Be dispensed into by the substratum prepared in the serum bottle of 100mL capacity, liquid amount is 30mL.Sealing, be filled with the nitrogen 5min of 99.999% with the speed of 2L/min after, at 121 DEG C of high-temperature sterilization 20min.After sterilizing, be cooled to about 40 DEG C, then add the micro-storage liquid of 0.1ml, and Na
2moO
42H
2o storage liquid and CuCl
22H
2o storage liquid, completes culture system and builds.Access is cultured to the Clostridiumcarboxidivorans bacterial strain P7 nutrient solution 3ml of logarithmic phase, then with 2L/min flow velocity, 0.2bar pressure durations passes into the (gas composition: 50%CO, 35%CO of analog synthesis gas
2, 15%H
2) balance 5min, be forced into 0.2MPa sealing.At 37 degree, shaking culture under 180rpm, tunning detection is carried out in sampling at regular intervals.
Embodiment 1:
In basic culture solution, do not add molybdenum acid ion.Through the constant-temperature shaking culture of 4 days, sampling in every 24 hours detected microorganism growth situation; Get the sample of the 4th day, utilize vapor-phase chromatography to detect the concentration of ethanol, propyl carbinol and n-hexyl alcohol in fermented liquid.Bacterial strain reaches 0.96 at the 4th day OD600, within the 4th day, starts to enter decline phase (Fig. 1).The production concentration of the 4th day ethanol, propyl carbinol and n-hexyl alcohol is respectively 0.24, and 0.16 and 0.06g/L (Fig. 2).
Embodiment 2:
In basic culture solution, adding final concentration is the molybdenum acid ion of 0.1mg/L.Through the constant-temperature shaking culture of 4 days, sampling in every 24 hours detected microorganism growth situation; Get the sample of the 4th day, utilize vapor-phase chromatography to detect the concentration of ethanol, propyl carbinol and n-hexyl alcohol in fermented liquid.Bacterial strain reaches 0.79 at the 3rd day OD600, within the 4th day, starts to enter decline phase (Fig. 3).Within 4th day, ethanol production is 0.07g/L, propyl carbinol and n-hexyl alcohol cannot be detected, and the growing state of bacterium is also poor simultaneously.
Embodiment 3:
In basic culture solution, adding final concentration is the molybdenum acid ion of 1mg/L.Through the constant-temperature shaking culture of 4 days, sampling in every 24 hours detected microorganism growth situation; Get the sample of the 4th day, utilize vapor-phase chromatography to detect the concentration of ethanol, propyl carbinol and n-hexyl alcohol in fermented liquid.The concentration that bacterial strain is the highest can only reach OD600=0.35 (Fig. 4).Any alcohols material cannot be detected in tunning.
Embodiment 4:
Use other minimal mediums instead, do not add molybdenum acid ion.Through the constant-temperature shaking culture of 4 days, sampling in every 24 hours detected microorganism growth situation; Get the sample of the 4th day, utilize vapor-phase chromatography to detect the concentration of ethanol, propyl carbinol and n-hexyl alcohol in fermented liquid.Bacterial strain reaches 0.92 at the 4th day OD600, within the 4th day, starts to enter decline phase (Fig. 5).The production concentration of the 4th day ethanol, propyl carbinol and n-hexyl alcohol is respectively 0.21, and 0.15 and 0.07g/L (Fig. 6).
Claims (8)
1. improve a method for anaerobism food gas fermentable efficiency, it is characterized in that, described method is that the final concentration of molybdenum acid ion in the substratum being used in fermentation is not more than 0.1mg/L.
2. the method for claim 1, is characterized in that, described method does not add molybdenum acid ion in the substratum for fermenting.
3. the method for claim 1, is characterized in that, described anaerobism food gas microorganism is anaerobism food gas clostridium.
4. the substratum of method use described in a claim 1, it is characterized in that, described substratum includes the mineralsolutionstock of 2%-4%, the yeast extract of 0.5%, the morpholino b acid of 0.5%, the reducingagentStock of 0.5%, the micro-storage liquid of 0.1%, and in substratum, the final concentration of molybdenum acid ion is not more than 0.1mg/L.
5. substratum as claimed in claim 4, it is characterized in that, described Mineralsolutionstock consists of 8% sodium-chlor, 10% ammonium chloride, 1% Repone K, the potassium primary phosphate of 1%, the magnesium sulfate of 2% and the calcium chloride of 0.4%.
6. substratum as claimed in claim 4, is characterized in that, described ReducingAgentStock consist of 0.9% sodium hydroxide, the L-cysteine hydrochloride of 4% and the nine water cure sodium of 4%.
7. substratum as claimed in claim 4, it is characterized in that, described liquid microelement storage liquid consist of 1% nitrilotriacetic acid(NTA), the magnesium sulfate of 0.5%, the ferrous ammonium sulphate of 0.4%, the cobalt chloride of 0.1%, the zinc sulfate of 0.1%, the nickelous chloride of 0.01%, the sodium selenate of 0.01% and the sodium wolframate of 0.01%.
8. substratum as claimed in claim 4, it is characterized in that, described Sodium orthomolybdate storage liquid is the Sodium orthomolybdate of 0.01%.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106148426A (en) * | 2016-09-19 | 2016-11-23 | 中国科学院天津工业生物技术研究所 | A kind of containing molybdenum, zinc, the cultivating system eating gas anaerobe fermentation product alcohol of nickel and cultural method |
| CN107779477A (en) * | 2016-08-25 | 2018-03-09 | 中国科学院天津工业生物技术研究所 | It is a kind of to eat cultivating system and cultural method of the gas anaerobic bacteria using synthesis gas fermentation production alcohol |
| CN107779478A (en) * | 2016-08-25 | 2018-03-09 | 中国科学院天津工业生物技术研究所 | It is a kind of to eat cultivating system and cultural method of the gas anaerobic bacteria using synthesis gas fermentation production alcohol |
| CN107779476A (en) * | 2016-08-25 | 2018-03-09 | 中国科学院天津工业生物技术研究所 | A kind of method that hexanol is produced using synthesis gas cold fermentation |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107779477A (en) * | 2016-08-25 | 2018-03-09 | 中国科学院天津工业生物技术研究所 | It is a kind of to eat cultivating system and cultural method of the gas anaerobic bacteria using synthesis gas fermentation production alcohol |
| CN107779478A (en) * | 2016-08-25 | 2018-03-09 | 中国科学院天津工业生物技术研究所 | It is a kind of to eat cultivating system and cultural method of the gas anaerobic bacteria using synthesis gas fermentation production alcohol |
| CN107779476A (en) * | 2016-08-25 | 2018-03-09 | 中国科学院天津工业生物技术研究所 | A kind of method that hexanol is produced using synthesis gas cold fermentation |
| CN107779476B (en) * | 2016-08-25 | 2021-10-12 | 中国科学院天津工业生物技术研究所 | Method for producing hexanol by low-temperature fermentation of synthesis gas |
| CN107779477B (en) * | 2016-08-25 | 2021-10-12 | 中国科学院天津工业生物技术研究所 | Culture system and culture method for producing alcohol by fermenting synthesis gas by using gas-feeding anaerobic bacteria |
| CN106148426A (en) * | 2016-09-19 | 2016-11-23 | 中国科学院天津工业生物技术研究所 | A kind of containing molybdenum, zinc, the cultivating system eating gas anaerobe fermentation product alcohol of nickel and cultural method |
| CN106148426B (en) * | 2016-09-19 | 2019-09-06 | 中国科学院天津工业生物技术研究所 | It is a kind of containing molybdenum, zinc, nickel food gas anaerobe fermentation produce alcohol cultivating system and cultural method |
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