CN107043792A - A kind of high temperature bacterium and the new technology of mesophile co-fermentation synthesis gas producing and ethanol - Google Patents
A kind of high temperature bacterium and the new technology of mesophile co-fermentation synthesis gas producing and ethanol Download PDFInfo
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Abstract
The invention discloses a kind of high temperature bacterium and the novel process of normal temperature bacterium co-fermentation synthesis gas producing and ethanol, comprise the following steps：(1)：The forming gas that gasification burner is generated is cooled to 50 DEG C~80 DEG C, and then high temperature bacterium and high temperature bacteria fermentation culture medium are placed in forming gas environment, and culture high temperature bacterium grows to the constant state of stable yield of tunning in the fermentation medium；(2)：When gas temperature to be synthesized drops to 37 DEG C, mesophile and first stage mesophile fermentation medium are added, the culture of first stage is carried out to mesophile, mesophile is grown to the constant state of stable yield of tunning in warm bacteria fermentation culture medium in the first phase；(3)：The mesophile that stable state is grown in step (2) is carried out to the culture of second stage, the nitrogen concentration of incubation second stage mesophile fermentation medium is controlled to be less than the first stage, and organic carbon source is removed, while controlling the pH value in incubation to be 4.0~4.5.
Description
【Technical field】
The invention belongs to field of biological energy source, more particularly to a kind of high temperature bacterium and mesophile co-fermentation synthesis gas producing and ethanol
New technology.
【Background technology】
Synthesis gas is the mixed gas produced by the oxidation of the biomass portions such as lignocellulosic and pyrolytic, it main
Composition is CO, H2And CO2, also containing a small amount of CH4With some sulphur, nitrogen compound.Synthesis gas is the abundant and cheap life of a class
Thing is processed raw material, and various useful fuels and chemicals, such as ethanol, acetic acid, butanol and butyric acid can be switched to by anaerobic fermentation
Deng.If using synthesis gas fermentation technique, first all biological matter (including lignin and difficult degradation part) gasification is converted into
Synthesis gas, then by synthesis gas fermentation be ethanol, with regard to can avoid lignocellulosic acid, enzyme hydrolysis technology barrier, overcome traditional life
The defect that lignin can not be fully utilized in thing conversion process.Biomass, discarded object and some cannot be used for direct fermentation
Raw material conversion on, synthesis gas fermentation will play a significant role.Outside degassifying synthesis gas, some industrial waste gases rich in CO are (such as
Converter gas, calcium carbide furnace gas, yellow phosphorus furnace gas etc.) utilization of gas fermentative microorganism can also be synthesized and converted.
Alcohol fuel is produced by raw material of synthesis gas, always due to high cost of material, and is ignored by people.Synthesis
Gas fermentation will play an important role in those processing for being difficult to the biomass of direct fermentation, discarded object and residue.Due to this
The synthesis gas temperature that a little gasification substances are produced is very high, and thermophilic microorganism is more suitable for this production technology.Synthesis gas in process of production
Need by the way that after a heat recovery system cooling, bioreactor could be entered.If synthesis gas first passes through thermophilic microorganism
After fermentation, then by normal temperature microbial fermentation, energy utilization rate will be effectively improved.At present, although synthesis gas produces ethanol also not
It is the principal mode of current alcohol production, but as a redundancy technique, is necessary to study it.
The research of synthesis gas fermentation producing and ethanol technology is at the early-stage at home, due to CO, CO2And H2Dissolving in a liquid
Degree is very low, causes microorganism few to the uptake of gas, there are problems that alcohol yied.Current domestic and international researcher is used
Different control measures improve the yield of ethanol, are concentrated mainly on the regulation and control for fermentation condition, including culture medium excellent
Change, pH control, the regulation and control of gas transfer efficiency and control of gas component and each component partial pressure etc..So far, state
Interior main research C.autoethanogenum bacterial strains utilize the optimization of synthesis gas fermenting alcohol fermentation condition.Mainly it is fermented
Culture medium is improved, and the optimal nitrogen source of culture medium is obtained after optimization for yeast extract, and optimal carbon source is xylose.Application No.
CN104822836A, CN104822836A and CN104812904A Chinese patent, it is proposed that the operation of synthesis gas fermentation process
CO is reduced in method, synthesis gas fermentation2Discharge and the method for improving alcohol production rate.Application No. CN105087441A China is specially
Profit, a kind of composite flora of proposition and its application in synthesis gas ferments production alcohol；Application No. CN106399387A China
Patent propose a kind of mixed fermentation synthesis gas producing and ethanol nitrogen defect regulation and control method, but be all not involved with high temperature bacterium and
The research of normal temperature bacterium co-fermentation synthesis gas producing and ethanol.
In view of above-mentioned deficiency, the design people is actively subject to research and innovation, to create a kind of new synthesis gas hair
Ferment pattern, makes it with more the application value in industry.
【The content of the invention】
The purpose of the present invention is to propose to a kind of new synthesis gas fermentation pattern, the synthesis gas of gasification burner generation is by appropriate
Cooling, first passes through the fermentation of high temperature bacterium, then by the fermentation of mesophile, produces the organic solvents such as ethanol.
To solve the above problems, the technical solution adopted in the present invention is：A kind of high temperature bacterium and mesophile co-fermentation are closed
Into the new technology of gas producing and ethanol, comprise the following steps：(1)：The forming gas that gasification burner is generated is cooled to 50 DEG C~80 DEG C, so
High temperature bacterium and high temperature bacteria fermentation culture medium are placed in forming gas environment afterwards, culture high temperature bacterium grows in the fermentation medium
The constant state of the stable yield of tunning；
(2)：When gas temperature to be synthesized drops to 37 DEG C, mesophile and first stage mesophile fermentation medium are added, it is right
Mesophile carries out the culture of first stage, mesophile is grown to tunning in warm bacteria fermentation culture medium in the first phase
The constant state of stable yield；
(3)：The mesophile that stable state is grown in step (2) is carried out to the culture of second stage, incubation is controlled
The nitrogen concentration of second stage mesophile fermentation medium is less than the first stage, and removes organic carbon source, while controlling incubation
In pH value be 4.0~4.5.
In the present invention, as further illustrating, the high temperature bacterium described in step (1) is Acetogenium kivui,
Carboxydocellas poropducens, Clostridium thermocellum, Desulfotomaculum
Thermobenzoicum sub sp themosyntrophicum, Moorella thermoacetica, Moorella
It is any in sp.HUC22-1, Moorella thermoautotrophica and Desulfotamaculum kuznetsovii
It is a kind of.
In the present invention, as further illustrating, the mesophile described in step (2) is Clostridium
Ljungdahlii, C.autoethanogenum, C.carboxidivorans, Butyribacterium
Any one in methylotrophicum, C.ragsdalei and Alkalibaculum bacchi.
In the present invention, as further illustrating, the high temperature bacteria fermentation culture medium described in step (1) includes consisting of：
Every liter as follows：NH4CL 1g/L, MgCL2·6H2O 0.16g/L, CaCL2·2H2O 0.1g/L, KCL 0.33g/L, KH2PO4
0.5g/L, 10mL trace element solution, 10mL vitamin solutions, 1.0g dusty yeasts, 2.0g peptones, 0.5g cysteine salts
Acid, 0.5mL mass fractions are 0.1% resazurin solution, and 5.0g sucrose, pH is 6.8.
In the present invention, as further illustrating, the first stage mesophile fermentation medium described in step (2) includes：
Every liter as follows：10mL inorganic salt solutions, 10mL trace element solutions, 10mL vitamin solutions, 1.0g dusty yeasts, 2.0g albumen
Peptone, 0.5g cysteine hydrochloric acid, 5.0g morpholino b acids, 0.5mL mass fractions be 0.1% resazurin solution, 5.0g fructose,
PH is 6.0.
In the present invention, as further illustrating, the second stage mesophile fermentation medium described in step (3) includes：
Every liter as follows：10mL inorganic salt solutions, 10mL trace element solutions, 10mL vitamin solutions, 0.5g dusty yeasts, 0.5g albumen
Peptone, 0.5g cysteine hydrochloric acid, 5.0g morpholino b acids, 0.5mL mass fractions are 0.1% resazurin solution, and pH is 6.0.
In the present invention, as further illustrating, trace element solution composition is as follows：NTA 2g/L,
MnCL24H2O1.3g/L, CoCL2·6H2O 0.2g/L, ZnSO4·7H2O 0.2g/L, FeCL3·6H20 0.4g/L,
CuCL2·2H2O 0.02g/L, NiCL2·6H2O 0.02g/L, Na2MoO4·2H2O 0.02g/L and Na2WO4·2H2O
0.025g/L。
In the present invention, as further illustrating, microbial solution composition is as follows：Biotin 2mg/L, folic acid 2mg/L, dimension
Raw element B6 10mg/L, riboflavin 5mg/L, vitamin B1 5mg/L, nicotinic acid 5mg/L, calcium pantothenate 5mg/L, vitamin B12 5mg/
L and p-aminobenzoic acid 5mg/L.
In the present invention, as further illustrating, inorganic salt solution composition is as follows：NaCL 80g/L,NH4CL 100g/L,
KCL1g/L, KH2PO410g/L, MgSO4·7H2O 20g/L and CaCL2·2H2O 4g/L。
In the present invention, as further illustrating, added in the second stage mesophile fermentation medium described in step (3)
BESA, coban and butyl rubber particle.
In the present invention, as further illustrating, the Organic carbon source described in step (3) is fructose or xylose.
In the present invention, as further illustrating, batch fermentation research is carried out using aluminium foil gas sampling bag.
The invention has the advantages that：
1. the present invention consider synthesis gas from the synthesis gas temperature that gasification burner comes out all compare high the characteristics of, now high temperature bacterium is not
Excessive cooling is needed just to participate in reaction, own metabolism is active, reaction efficiency is high while high temperature bacterium has, dirty miscellaneous few etc. excellent
Point；The synthesis gas with the source such as lignin and cellulose can be used to ferment for the technology used in the present invention means, and produce new energy
Ethanol causes changing waste into resources, and reduces CO2The discharge of isothermal chamber gas, meets continuable development principle；Fermented for synthesis gas
The heavy industrialization of producing and ethanol, which is applied, provides important theory value and actual directive significance.
2. the present invention is outside degassifying synthesis gas, some industrial waste gases rich in CO can also be had by this scheme
Organic solvent, the present invention is more attractive to the industrial waste gas rich in CO.High temperature bacterium in the first stage can be with
It is that sole carbon source generates H using CO2And CO2, the gas fraction after fermentation is than about CO, CO2And H2Volume ratio be 4：3：3, fit
Close the growth of various synthesis gas fermentation mesophiles；And high temperature bacterium just loses reaction vigor automatically in middle tender feeling condition, will not be with
Warm bacterium produces competition to nutriment, and itself can serve as nutriment；So this technological means can be production
New energy provides new thinking.
【Brief description of the drawings】
Fig. 1 is the yield of synthesis gas fermenting alcohol in embodiment 1；
Fig. 2 is the yield of synthesis gas fermenting alcohol in embodiment 2；
Fig. 3 is the yield of synthesis gas fermenting alcohol in embodiment 3；
Fig. 4 is the yield of synthesis gas fermenting alcohol in embodiment 4；
Fig. 5 is the yield of synthesis gas fermenting alcohol in embodiment 5.
【Embodiment】
Strain source：High temperature bacterium Acetogenium kivui, Carboxydocellas poropducens,
Clostridium thermocellum, Desulfotomaculum thermobenzoicum sub sp
Themosyntrophicum, Moorella thermoacetica, Moorella sp.HUC22-1, Moorella
Thermoautotrophica and Desulfotamaculum kuznetsovii；Mesophile Clostridium
Ljungdahlii, C.autoethanogenum, C.carboxidivorans, Butyribacterium
Methylotrophicum, C.ragsdalei and Alkalibaculum bacchi.High temperature bacterium and mesophile strain are by be wide
Western DSMZ of the academy of sciences provides.
Embodiment 1：
A kind of high temperature bacterium and the new technology of mesophile co-fermentation synthesis gas producing and ethanol, comprise the following steps：
(1) 5mL high temperature bacterial strain seed liquor Acetogenium kivui are added to 500mL high temperature bacteria fermentation culture medium
In, high temperature bacteria fermentation culture medium constitute every liter it is as follows：NH4CL 1g/L, MgCL2·6H2O 0.16g/L, CaCL2·2H2O
0.1g/L, KCL 0.33g/L, KH2PO40.5g/L, 10mL trace element solution, 10mL vitamin solutions, 1.0g dusty yeasts,
2.0g peptones, 0.5g cysteine hydrochloric acid, 0.5mL mass fractions are 0.1% resazurin solution, 5.0g sucrose, pH6.8.
CO gases are filled with to a standard atmospheric pressure, adjusts and first stage culture is carried out at pH=6.8,50 DEG C, are grown after cultivating 3 days
To stable state；
(2) first stage fluid nutrient medium is cooled to 37 DEG C, is directly inoculated with mesophile strain seed liquor Clostridium
Ljungdahlii is into first stage mesophile fermentation medium, and first stage mesophile fermentation medium composition is：Every liter such as
Under：10mL inorganic salt solutions, 10mL trace element solutions, 10mL vitamin solutions, 1.0g dusty yeasts, 2.0g peptones, 0.5g
Cysteine hydrochloric acid, 5.0g morpholino b acids, 0.5mL mass fractions are 0.1% resazurin solution, 5.0g fructose, and pH is
6.0.Now CO, CO in gas pouch2And H2The Volume fraction of gas is 4：3：3, it is an atmospheric pressure, pH value to maintain pressure
For 6.0.Homoacetogenic bacteria grows to stable state after cultivating 5 days；
(3) take the enriched substance strain after 20mL steps (2) culture into 500mL aluminium foil gas sampling bags and contain low concentration
The second stage mesophile fermentation medium of nitrogen source：10mL inorganic salt solutions, 10mL trace element solutions, 10mL vitamins are molten
Liquid, 0.5g yeast extracts, 0.5g peptones, 0.5g cysteine hydrochloric acid, 0.5mL mass fractions are 0.1% resazurin solution, pH
For 6.0,0.2gBESA, 0.1g coban, 0.1g butyl rubber particles.Fructose Organic carbon source is removed, 37 DEG C of constant temperature trainings are placed in
Support in case and cultivate 20 days.Now CO, CO in gas pouch2And H2The Volume fraction of gas is 4：3：3, it is one to maintain pressure
Atmospheric pressure, it is 4.5 to maintain pH.Utilize gas chromatographic measurement gas consumption situation and the organic product condition of production.
It is computed, the concentration of alcohol of generation is 3.9g/L.
Embodiment 2：
A kind of high temperature bacterium and the new technology of mesophile co-fermentation synthesis gas producing and ethanol, comprise the following steps：
(1) 5mL high temperature bacterial strain seed liquor Carboxydocella sporoproducens are added to 500mL high temperature bacterium
In fermentation medium, high temperature bacteria fermentation culture medium constitute every liter it is as follows：NH4CL 1g/L, MgCL2·6H2O 0.16g/L,
CaCL2·2H2O, 0.1g/L KCL, 0.33g/L KH2PO40.5g/L, 10mL trace element solution, 10mL vitamin solutions,
1.0g dusty yeasts, 2.0g peptones, 0.5g cysteine hydrochloric acid, 0.5mL mass fractions are 0.1% resazurin solution, 5.0g
Sucrose, pH 6.8.Now CO, CO in gas pouch2Volume ratio is 4：1, maintain to a standard atmospheric pressure, adjust pH=6.8,
First stage culture is carried out at 60 DEG C, stable state is grown to after cultivating 3 days；
(2) first stage fluid nutrient medium is cooled to 37 DEG C, is directly inoculated with mesophile strain seed liquor
C.autoethanogenum is into first stage mesophile fermentation medium, and first stage mesophile fermentation medium composition is：
Every liter as follows：10mL inorganic salt solutions, 10mL trace element solutions, 10mL vitamin solutions, 1.0g dusty yeasts, 2.0g albumen
Peptone, 0.5g cysteine hydrochloric acid, 5.0g morpholino b acids, the resazurins of 0.5mL 0.1%, 5.0g fructose, pH 6.0.Now gas
CO, CO in bag2And H2The Volume fraction of gas is 4：3：3, it is an atmospheric pressure to maintain pressure, and pH value is 6.0.Treat culture 5
Homoacetogenic bacteria grows to stable state after it；
(3) the enriched substance strain after 40mL steps (2) culture is taken into 1000mL aluminium foil gas sampling bags and containing low dense
Spend the second stage mesophile fermentation medium of nitrogen source：10mL inorganic salt solutions, 10mL trace element solutions, 10mL vitamins are molten
Liquid, 0.5g yeast extracts, 0.5g peptones, 0.5g cysteine hydrochloric acid, 0.5mL mass fractions are 0.1% resazurin solution, pH
For 6.0,0.2gBESA, 0.1g coban, 0.2g butyl rubber particles.Xylose Organic carbon source is removed, 37 DEG C of constant temperature trainings are placed in
Support in case and cultivate 30 days.Now CO, CO in gas pouch2And H2The Volume fraction of gas is 4：3：3, it is one to maintain pressure
Atmospheric pressure, it is 4.0 to maintain pH.Utilize gas chromatographic measurement gas consumption situation and the organic product condition of production.
It is computed, the concentration of alcohol of generation is 4.7g/L.
Embodiment 3：
A kind of high temperature bacterium and the new technology of mesophile co-fermentation synthesis gas producing and ethanol, comprise the following steps：
(1) 5mL high temperature bacterial strain seed liquor Clostridium thermocellum are added to the fermentation training of 500mL high temperature bacterium
Support base in, high temperature bacteria fermentation culture medium constitute every liter it is as follows：NH4CL 1g/L, MgCL2·6H2O 0.16g/L, CaCL2·2H2O
0.1g/L, KCL 0.33g/L, KH2PO40.5g/L, 10mL trace element solution, 10mL vitamin solutions, 1.0g dusty yeasts,
2.0g peptones, 0.5g cysteine hydrochloric acid, 0.5mL mass fractions are 0.1% resazurin solution, 5.0g sucrose, and pH is
6.8.CO gases are filled with to a standard atmospheric pressure, adjusts and first stage culture is carried out at pH=6.8,68 DEG C, after after culture 3 days
Grow to stable state；
(2) first stage fluid nutrient medium is cooled to 37 DEG C, is directly inoculated with mesophile strain seed liquor
C.carboxidivorans is into first stage mesophile fermentation medium, and first stage mesophile fermentation medium composition is：
Every liter as follows：10mL inorganic salt solutions, 10mL trace element solutions, 10mL vitamin solutions, 1.0g dusty yeasts, 2.0g albumen
Peptone, 0.5g cysteine hydrochloric acid, 5.0g morpholino b acids, 0.5mL mass fractions be 0.1% resazurin solution, 5.0g fructose,
PH is 6.0.Now CO, CO in gas pouch2And H2The Volume fraction of gas is 4：3：3, it is an atmospheric pressure to maintain pressure,
PH value maintains 6.0.Homoacetogenic bacteria grows to stable state after cultivating 5 days；
(3) the enriched substance strain after 40mL steps (2) culture is taken into 1000mL aluminium foil gas sampling bags and containing low dense
Spend the second stage mesophile fermentation medium of nitrogen source：10mL inorganic salt solutions, 10mL trace element solutions, 10mL vitamins are molten
Liquid, 0.5g yeast extracts, 0.5g peptones, 0.5g cysteine hydrochloric acid, 0.5mL mass fractions are 0.1% resazurin solution, pH
For 4.3,0.1gBESA, 0.2g coban, 0.1g butyl rubber particles.Fructose Organic carbon source is removed, 37 DEG C of constant temperature trainings are placed in
Support in case and cultivate 30 days.Now CO, CO in gas pouch2And H2The Volume fraction of gas is 4：3：3, it is one to maintain pressure
Atmospheric pressure, it is 6.0 to maintain pH.Utilize gas chromatographic measurement gas consumption situation and the organic product condition of production.
It is computed, the concentration of alcohol of generation is 7.3g/L.
Embodiment 4：
A kind of high temperature bacterium and the new technology of mesophile co-fermentation synthesis gas producing and ethanol, comprise the following steps：
(1) by 5mL high temperature bacterial strain seed liquor Desulfotomaculum thermobenzoicum sub sp
Themosyntrophicum is added in 500mL high temperature bacteria fermentation culture mediums, high temperature bacteria fermentation culture medium constitute every liter it is as follows：
NH4CL 1g/L, MgCL2·6H2O 0.16g/L, CaCL2·2H2O 0.1g/L, KCL 0.33g/L, KH2PO40.5g/L,
10mL trace element solutions, 10mL vitamin solutions, 1.0g dusty yeasts, 2.0g peptones, 0.5g cysteine hydrochloric acid, 0.5mL
Mass fraction is 0.1% resazurin solution, and 5.0g sucrose, pH is 6.8.It is filled with CO, CO2Gas volume ratio is 4：1 mixing
Gas is adjusted and first stage culture is carried out at pH=6.8,70 DEG C to a standard atmospheric pressure, and stabilization is grown to after cultivating 3 days
State；
(2) first stage fluid nutrient medium is cooled to 37 DEG C, is directly inoculated with mesophile strain seed liquor
Butyribacterium methylotrophicum are into first stage mesophile fermentation medium, first stage mesophile hair
Ferment culture medium is constituted：10mL inorganic salt solutions, 10mL trace element solutions, 10mL vitamin solutions, 1.0g yeast extracts, 2.0g
Peptone, 0.5g cysteine hydrochloric acid, 0.5mL mass fractions are 0.1% resazurin solution, and 5.0g xyloses, pH is 6.0.This
When gas pouch in CO, CO2And H2The Volume fraction of gas is 4：3：3, it is an atmospheric pressure to maintain pressure, and pH value is maintained
6.0.Grow to stable state after homoacetogenic bacteria after culture 6 days and enter third step；
(3) take the enriched substance strain after 10mL steps (2) culture into 500mL aluminium foil gas sampling bags and contain low concentration
The second stage mesophile fermentation medium of nitrogen source：10mL inorganic salt solutions, 10mL trace element solutions, 10mL vitamins are molten
Liquid, 0.5g yeast extracts, 0.5g peptones, 0.5g cysteine hydrochloric acid, 0.5mL mass fractions are 0.1% resazurin solution, pH
For 6.0,0.1gBESA, 0.1g coban, 0.1g butyl rubber particles.Xylose Organic carbon source is removed, 37 DEG C of constant temperature trainings are placed in
Support in case and cultivate 15 days.Now CO, CO in gas pouch2And H2The Volume fraction of gas is 4：3：3, it is one to maintain pressure
Atmospheric pressure, and maintain pH to be 4.2.Utilize gas chromatographic measurement gas consumption situation and the organic product condition of production.
It is computed, the concentration of alcohol of generation is 3.2g/L.
Embodiment 5：
A kind of high temperature bacterium and the new technology of mesophile co-fermentation synthesis gas producing and ethanol, comprise the following steps：
(1) 5mL high temperature bacterial strain seed liquor Moorella thermoacetica are added to 500mL high temperature bacterium fermented and cultureds
In base, CO, CO are filled with2Gas volume ratio is 4：1 mixed gas is adjusted and entered at pH=6.8,60 DEG C to a standard atmospheric pressure
The row first stage is cultivated, and stable state is grown to after cultivating 6 days；
(2) first stage fluid nutrient medium is cooled to 37 DEG C, directly inoculation mesophile strain seed liquor C.ragsdalei is arrived
In first stage mesophile fermentation medium, first stage mesophile fermentation medium composition is：10mL inorganic salt solutions, 10mL
Trace element solution, 10mL vitamin solutions, 1.0g yeast extracts, 2.0g peptones, 0.5g cysteine hydrochloric acid, 0.5mL mass
Number is 0.1% resazurin solution, and 5.0g xyloses, pH is 6.0.It is filled with CO2Second stage pressure stability is maintained in a mark
Quasi- atmospheric pressure, pH value maintains 6.0.Grow to stable state after homoacetogenic bacteria after culture 6 days and enter third step；
(3) the enriched substance strain after 10mL steps (2) culture is taken into 1000mL aluminium foil gas sampling bags and containing low dense
Spend the second stage mesophile fermentation medium of nitrogen source：10mL inorganic salt solutions, 10mL trace element solutions, 10mL vitamins are molten
Liquid, 0.5g yeast extracts, 0.5g peptones, 0.5g cysteine hydrochloric acid, 0.5mL mass fractions are 0.1% resazurin solution, pH
For 6.0,0.2gBESA, 0.2g coban, 0.2g butyl rubber particles.Fructose Organic carbon source is removed, 37 DEG C of constant temperature trainings are placed in
Support in case and cultivate 20 days.Now CO, CO in gas pouch2And H2The Volume fraction of gas is 4：3：3, it is one to maintain pressure
Atmospheric pressure, and maintain pH to be 4.1.Utilize gas chromatographic measurement gas consumption situation and the organic product condition of production.
It is computed, the concentration of alcohol of generation is 3.9g/L.
Embodiment 6：
A kind of high temperature bacterium and the new technology of mesophile co-fermentation synthesis gas producing and ethanol, comprise the following steps：
(1) 5mL high temperature bacterial strain seed liquor Moorella sp.HUC22-1 are added to 500mL high temperature bacteria fermentation culture mediums
In, it is filled with CO, CO2Gas volume ratio is 4：1 mixed gas is adjusted and carried out at pH=6.8,75 DEG C to a standard atmospheric pressure
First stage is cultivated, and stable state is grown to after cultivating 6 days；
(2) first stage fluid nutrient medium is cooled to 37 DEG C, is directly inoculated with mesophile strain seed liquor Alkalibaculum
Bacchi is into first stage mesophile fermentation medium, and first stage mesophile fermentation medium composition is：10mL inorganic salts
Solution, 10mL trace element solutions, 10mL vitamin solutions, 1.0g yeast extracts, 2.0g peptones, 0.5g cysteine hydrochloric acid,
0.5mL mass fractions are 0.1% resazurin solution, and 5.0g xyloses, pH is 6.0.It is filled with CO2Maintain second stage pressure stability
In a standard atmospheric pressure, pH value maintains 6.0.Homoacetogenic bacteria grows to stable state into the 3rd step after cultivating 6 days
Suddenly；
(3) the enriched substance strain after 10mL steps (2) culture is taken into 1000mL aluminium foil gas sampling bags and containing low dense
Spend the second stage mesophile fermentation medium of nitrogen source：10mL inorganic salt solutions, 10mL trace element solutions, 10mL vitamins are molten
Liquid, 0.5g yeast extracts, 0.5g peptones, 0.5g cysteine hydrochloric acid, 0.5mL mass fractions are 0.1% resazurin solution, pH
For 6.0,0.1gBESA, 0.1g coban, 0.1g butyl rubber particles.Xylose Organic carbon source is removed, 37 DEG C of constant temperature trainings are placed in
Support in case and cultivate 20 days.Now CO, CO in gas pouch2And H2The Volume fraction of gas is 4：3：3, it is one to maintain pressure
Atmospheric pressure, and maintain pH to be 4.4.Utilize gas chromatographic measurement gas consumption situation and the organic product condition of production.
It is computed, the concentration of alcohol of generation is 5.2g/L.
Embodiment 7：
A kind of high temperature bacterium and the new technology of mesophile co-fermentation synthesis gas producing and ethanol, comprise the following steps：
(1) 5mL high temperature bacterial strain seed liquor Moorella thermoautotrophica are added to 500mL high temperature bacterium hair
In ferment culture medium, CO, CO are filled with2Gas volume ratio is 4：1 mixed gas adjusts pH=6.8,63 to a standard atmospheric pressure
First stage culture is carried out at DEG C, stable state is grown to after cultivating 6 days；
(2) first stage fluid nutrient medium is cooled to 37 DEG C, directly inoculation mesophile strain seed liquor C.ragsdalei is arrived
In first stage mesophile fermentation medium, first stage mesophile fermentation medium composition is：10mL inorganic salt solutions, 10mL
Trace element solution, 10mL vitamin solutions, 1.0g yeast extracts, 2.0g peptones, 0.5g cysteine hydrochloric acid, 0.5mL mass
Number is 0.1% resazurin solution, and 5.0g xyloses, pH is 6.0.It is filled with CO2Second stage pressure stability is maintained in a mark
Quasi- atmospheric pressure, pH value maintains 6.0.Grow to stable state after homoacetogenic bacteria after culture 6 days and enter third step；
(3) the enriched substance strain after 10mL steps (2) culture is taken into 1000mL aluminium foil gas sampling bags and containing low dense
Spend the second stage mesophile fermentation medium of nitrogen source：10mL inorganic salt solutions, 10mL trace element solutions, 10mL vitamins are molten
Liquid, 0.5g yeast extracts, 0.5g peptones, 0.5g cysteine hydrochloric acid, 0.5mL mass fractions are 0.1% resazurin solution, pH
For 6.0,0.1gBESA, 0.2g coban, 0.2g butyl rubber particles.Fructose Organic carbon source is removed, 37 DEG C of constant temperature trainings are placed in
Support in case and cultivate 20 days.Now CO, CO in gas pouch2And H2The Volume fraction of gas is 4：3：3, it is one to maintain pressure
Atmospheric pressure, and maintain pH to be 4.5.Utilize gas chromatographic measurement gas consumption situation and the organic product condition of production.
It is computed, the concentration of alcohol of generation is 4.5g/L.
Embodiment 8：
A kind of high temperature bacterium and the new technology of mesophile co-fermentation synthesis gas producing and ethanol, comprise the following steps：
(1) 5mL high temperature bacterial strain seed liquor Alkalibaculum bacchi are added to 500mL high temperature bacteria fermentation culture mediums
In, it is filled with CO, CO2Gas volume ratio is 4：1 mixed gas is adjusted and carried out at pH=6.8,80 DEG C to a standard atmospheric pressure
First stage is cultivated, and stable state is grown to after cultivating 6 days；
(2) first stage fluid nutrient medium is cooled to 37 DEG C, directly inoculation mesophile strain seed liquor C.ragsdalei is arrived
In first stage mesophile fermentation medium, first stage mesophile fermentation medium composition is：10mL inorganic salt solutions, 10mL
Trace element solution, 10mL vitamin solutions, 1.0g yeast extracts, 2.0g peptones, 0.5g cysteine hydrochloric acid, 0.5mL mass
Number is 0.1% resazurin solution, and 5.0g xyloses, pH is 6.0.It is filled with CO2Second stage pressure stability is maintained in a mark
Quasi- atmospheric pressure, pH value maintains 6.0.Grow to stable state after homoacetogenic bacteria after culture 6 days and enter third step；
(3) the enriched substance strain after 10mL steps (2) culture is taken into 1000mL aluminium foil gas sampling bags and containing low dense
Spend the second stage mesophile fermentation medium of nitrogen source：10mL inorganic salt solutions, 10mL trace element solutions, 10mL vitamins are molten
Liquid, 0.5g yeast extracts, 0.5g peptones, 0.5g cysteine hydrochloric acid, 0.5mL mass fractions are 0.1% resazurin solution, pH
For 6.0,0.2gBESA, 0.1g coban, 0.1g butyl rubber particles.Xylose Organic carbon source is removed, 37 DEG C of constant temperature trainings are placed in
Support in case and cultivate 20 days.Now CO, CO in gas pouch2And H2The Volume fraction of gas is 4：3：3, it is one to maintain pressure
Atmospheric pressure, and maintain pH to be 4.3.Utilize gas chromatographic measurement gas consumption situation and the organic product condition of production.
It is computed, the concentration of alcohol of generation is 4.7g/L.
Described above is the detailed description for the present invention preferably possible embodiments, but embodiment is not limited to this hair
The equal change or modification change completed under bright patent claim, the technical spirit suggested by all present invention, all should belong to
Cover the scope of the claims in the present invention.
Claims (6)
1. a kind of high temperature bacterium and the new technology of mesophile co-fermentation synthesis gas producing and ethanol, it is characterised in that：Comprise the following steps：
(1)：The forming gas that gasification burner is generated is cooled to 50 DEG C~80 DEG C, then by high temperature bacterium and high temperature bacteria fermentation culture medium
It is placed in forming gas environment, culture high temperature bacterium grows to the constant shape of stable yield of tunning in the fermentation medium
State；
(2)：When gas temperature to be synthesized drops to 37 DEG C, mesophile and first stage mesophile fermentation medium, centering temperature are added
Bacterium carries out the culture of first stage, mesophile is grown to the yield of tunning in warm bacteria fermentation culture medium in the first phase
Stablize constant state；
(3)：The mesophile that stable state is grown in step (2) is carried out to the culture of second stage, incubation second is controlled
The nitrogen concentration of stage mesophile fermentation medium is less than the first stage, and removes organic carbon source, while controlling in incubation
PH value is 4.0~4.5.
2. a kind of high temperature bacterium according to claim 1 and the new technology of mesophile co-fermentation synthesis gas producing and ethanol, it is special
Levy and be：High temperature bacterium described in step (1) is Acetogenium kivui, Carboxydocellas poropducens,
Clostridium thermocellum, Desulfotomaculum thermobenzoicum sub sp
Themosyntrophicum, Moorella thermoacetica, Moorella sp.HUC22-1, Moorella
Any one in thermoautotrophica and Desulfotamaculum kuznetsovii.
3. a kind of high temperature bacterium according to claim 1 and the new technology of mesophile co-fermentation synthesis gas producing and ethanol, it is special
Levy and be：Mesophile described in step (2) is Clostridium ljungdahlii, C.autoethanogenum,
C.carboxidivorans, Butyribacterium methylotrophicum, C.ragsdalei and Alkalibaculum
Any one in bacchi.
4. a kind of high temperature bacterium according to claim 1 and the new technology of mesophile co-fermentation synthesis gas producing and ethanol, it is special
Levy and be：BESA, coban and butyl rubber are added in second stage mesophile fermentation medium described in step (3)
Grain.
5. a kind of high temperature bacterium according to claim 1 and the new technology of mesophile co-fermentation synthesis gas producing and ethanol, it is special
Levy and be：Organic carbon source described in step (3) is fructose or xylose.
6. a kind of high temperature bacterium according to claim 1 and the new technology of mesophile co-fermentation synthesis gas producing and ethanol, it is special
Levy and be：Batch fermentation research is carried out using aluminium foil gas sampling bag.
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