CN107964553A - Method for producing butanol - Google Patents
Method for producing butanol Download PDFInfo
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- CN107964553A CN107964553A CN201610912605.7A CN201610912605A CN107964553A CN 107964553 A CN107964553 A CN 107964553A CN 201610912605 A CN201610912605 A CN 201610912605A CN 107964553 A CN107964553 A CN 107964553A
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- Prior art keywords
- butanol
- clostridium
- method described
- fermentation
- bacterial strain
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
- C12P7/16—Butanols
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
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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
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
Abstract
The present invention discloses a kind of method for producing butanol, it includes：The bacterial strain of one Clostridial species is fixed on a fixation support；And make the bacterial strain through fixed Clostridial species continuously ferment, and make it that butanol is generated, wherein, butanol is to be recovered during continuously fermenting by using the air lift in situ of a carrier gas.
Description
Technical field
The invention relates to a kind of method for producing butanol, it includes：By the bacterial strain (a of a Clostridial species
Strain of Clostridium spp.) it is fixed on a fixation support (immobilization carrier)；And
Make the bacterial strain through fixed Clostridial species continuously ferment, and make it that butanol is generated, wherein, butanol is continuously to send out
During ferment by using the air lift in situ (in situ gas stripping) of a carrier gas (carrier gas) and by
Recycling.
Background technology
Butanol (butanol) has high-energy-density (high energy density), low-steam pressure (low vapour
Pressure), low polarity and the intersolubility (miscibility) with gasoline, thus it is used as fuel synergistic agent (fuel
Extender), and it is considered as one of following important energy source.
Butanol can be by using Clostridial species (Clostridium spp.) [for example, acetone fourth under an anaerobic condition
Alcohol clostridium (Clostridium acetobutylicum), Clostridium beijerinckii (Clostridium beijerinckii) and sugared second
The more Clostridium acetobutylicums (Clostridium saccharoperbutylacetonicum) of acid] bacterial strain come carry out fermentation reaction and by
Production.However, butanol has cytotoxicity, and when the butanol accumulation generated in fermentation reaction is more, the growth meeting of bacterial strain
It is suppressed, and then causes the attenuating of butanol yield.
In order to reduce the adverse effect caused by the accumulation of butanol, having studied is continuously fermented using immobilization
(immobilized continuous fermentation) or air lift (gas stripping) constantly remove butanol.Example
Such as, in Qureshi N.et al. (2004), Appl.Biochem.Biotechnol., 114:In 713-721, Qureshi N.
Et al. will be as sterilized clay brick particle (the clay brick of fixation support (immobilization carrier)
Particle) it is placed in a reactor, then activated Clostridium beijerinckii is seeded in the reactor, then by the reactor
Filling is with P2 culture mediums.Culture last 4 it is small when after, fresh P2 culture mediums are by lasting charging into the reactor, and this is anti-
The culture in device is answered constantly to be discharged, in order to be 0.32h one-1Dilution rate (dilution rate) under connected
Supervention ferment.And experimental result is found：The immobilization obtained butanol yield that continuously ferments is to be significantly better than Batch fermentation
(batch fermentation) institute tool person.
In Ezeji T.C.et al. (2003), WorldJ.Microbiol.Biotechnol., 19:In 595-603,
Clostridium beijerinckii is seeded in a fermentation tank containing P2 culture mediums and carries out Batch fermentation by Ezeji T.C. et al..Proceeding by
The 15th of Batch fermentation it is small when from, with the CO generated during the fermentation2And H2As carrier gas (carrier gas)
And air lift (in situ gas in situ are carried out to the fermentation culture medium in the fermentation tank under the flow velocity that one is 3LPM
Stripping), the butanol being extracted is collected by a condenser.And experimental result is found：Air lift, which can be lifted, to be criticized
The butanol yield of secondary fermentation and glucose utilization rate (glucose utilization).
In addition, in order to further improve the efficiency of air lift, study and removed the thalline in fermentation culture medium, it is then right
The fluid section of obtained not mycetome carries out air lift.For example, CN103555560B discloses a kind of acetone butanol fermentation coupling
Isolating and purifying the device for preparing butanol and the method that butanol is prepared using the device, this method is included：By a production of butanol
Bacterium, which is seeded in a stirring type bioreactor, ferments, and utilizes an immobilization device and a bacterium solution separator will
Thalline is isolated from fermentation culture medium, then carries out air lift to the fluid section of obtained not mycetome.In this part continent
In the embodiment of Patent Case, the gas stripping efficiency of this method is proved to be to be significantly better than the method institute without using bacterium solution separator
Tool person.
Master's thesis { title written by Wang Yinrong in chemical engineering research institute of TaiWan, China Chung Hsing University：" directly in fixation
Change bed body and carry out continous way immobilization acetone-butanol alcohol fermentation [the Direct in situ butanol that butanol in situ removes
removal on the packed bed during continuous and immobilized Acetone-Butanol-
Ethanol (ABE) fermentation] " } in, Wang Yinrong continuously ferments using through fixed clostridium acetobutylicum, and
And obtained fermentation culture medium is extracted using oleyl alcohol (oleyl alcohol) as extractant, to be contained
The oleyl alcohol extract of butanol, then the oleyl alcohol extract is to be stripped in an extraction tank by nitrogen.
However, these previous research need to use extra separating step and equipment and improve required cost, production is not applied
Practical application in industry.Therefore, if a kind of system with high butanol yield, ease of Use program and low cost can be developed
Journey, can be that we long for and reach.
The content of the invention
Then, the present invention provides a kind of method for producing butanol, it includes：
The bacterial strain of one Clostridial species is fixed on a fixation support；And
Make the bacterial strain through fixed Clostridial species continuously ferment, and make it that butanol is generated, wherein, butanol is
It is recovered during continuously fermenting by using the air lift in situ of a carrier gas.
Brief description of the drawings
Next with reference to the accompanying drawings and embodiments the present invention is described in detail, so the present invention is in above-mentioned and other mesh
And feature, can by with reference to following description, examine attached claims and adjoint schema with text and become apparent,
In attached drawing：
Fig. 1 is a schematic diagram for continuously fermenting reactor.
Embodiment
In order to simplify the butanol fermentation processing procedure for using the bacterial strain of Clostridial species to be carried out and improve process efficiency, apply
People has found through result of study of joining hands, to be continuously fermented through fixed bacterial strain and during continuously fermenting with air lift in situ
To recycle butanol, glucose utilization rate and butanol yield can be effectively improved.
Then, the present invention provides a kind of method for producing butanol, it includes：
The bacterial strain of one Clostridial species is fixed on a fixation support；And
Make the bacterial strain through fixed Clostridial species continuously ferment, and make it that butanol is generated, wherein, butanol is
It is recovered during continuously fermenting by using the air lift in situ of a carrier gas.
As used in this article, term " bacterial strains (strain of Clostridium spp.) of Clostridial species " is intended to
Cover all bacterial strains for belonging to Clostridial species that can produce butanol, be suitable for the invention the bacterial strain bag of Clostridial species
Include, but be not limited to：Clostridium acetobutylicum (Clostridium acetobutylicum), clostridium saccharoacetoperbutylicum
(Clostridium saccharoperbutylacetonicum), Clostridium beijerinckii (Clostridium beijerinckii),
Clostridium tyrobutyricum (Clostridium tyrobutyricum), clostridium butyricum (Clostridium butyricum), and it
Combination.In the preferred embodiment of the present invention, the bacterial strain of the Clostridial species is clostridium acetobutylicum.
As used in this article, term " fixed (immobilizing) " or " immobilization (immobilization) " mean
One microorganism to live is confined in specific an area of space or carrier, the fluid dynamic for whereby showing the Institute of Micro-biology
It is to be different from surrounding environment institute tool person to learn feature (hydrodynamic characteristic), and can keep the microorganism
Activity and be used repeatedly it.
According to the present invention, the immobilization of the bacterial strain of Clostridial species, which can use, to be familiar with this those skilled in the art and know in detail and usual
Technology carry out.In this regard, may be referred to, for example, Qureshi N.et al. (2004) (with above-mentioned) and Chinese platform
Master's thesis written by the Wang Yinrong of chemical engineering research institute of gulf Chung Hsing University (with above-mentioned).
It although it is understood that, the operating condition in relation to immobilization can be further with the bacterial strain of used Clostridial species
And the factor such as species, particle diameter and carrier filling rate of fixation support and changed, so as to up to causing optimal immobilization effect
Fruit.And the selection of these operating conditions is familiar with this those skilled in the art and can decide in its sole discretion to routine.
It is preferred that the fixation support is to be selected from following formed group：Clay brick (clay brick), zeolite
(zeolite), ceramic (ceramic), bone black (bone char), resin (resin), sawdust (wood chip), coke
(coke), rice husk (rice husk), cotton (cotton), cornstalk (corn stalk), calcium alginate (calcium
Alginate), silica gel (silica gel), and combinations thereof.In the preferred embodiment of the present invention, the immobilization
Carrier is clay brick.
According to the present invention, the particle diameter of the fixation support is fallen in the range of 0.15mm to 5mm.At one of the present invention
In preferred embodiment, the particle diameter of the fixation support is fallen in the range of 0.84mm to 2.38mm.
As used in this article, term " carrier filling rate (carrier filling rate) " means fixation support
Weight relative to the volume of the culture medium in fermentation tank ratio.According to the present invention, the carrier filling rate of the fixation support
It is to fall in the range of 20% (w/v, g/L) to 40% (w/v, g/L).In the preferred embodiment of the present invention, this is fixed
The carrier filling rate for changing carrier is 20% (w/v, g/L).
According to the present invention, the immobilization of the bacterial strain of the Clostridial species is under an anaerobic condition and a scope falls 30
DEG C last 30 minutes to 120 minutes to culture at the temperature in 40 DEG C and carried out.In the preferred embodiment of the present invention,
The immobilization of the bacterial strain of the Clostridial species is that culture lasts 30 minutes and carried out under an anaerobic condition and at 37 DEG C.
According to the present invention, before being continuously fermented can the Schilling bacterial strain through fixed Clostridial species carry out batch hair
Ferment, to improve the bacterium number of the bacterial strain through fixed Clostridial species.
According to the present invention, this, which continuously ferments, to be known in detail using this those skilled in the art is familiar with and usual technology carries out.
In this regard, may be referred to, for example, Qureshi N.et al. (2004) (with above-mentioned) and TaiWan, China Chung Hsing University chemistry
Master's thesis written by the Wang Yinrong of Graduate School of Engineering (with above-mentioned).
It although it is understood that, the operating condition in relation to continuously fermenting can be further with the bacterium of used Clostridial species
The factor such as formula of strain and culture medium and changed, so as to up to causing optimal ferment effect.And the selection of these operating conditions
Being familiar with this those skilled in the art can decide in its sole discretion to routine.
According to the present invention, which includes the carbon of the strain growth suitable for Clostridial species
Source (carbon source), nitrogen source (nitrogen source) and inorganic salt and other material.These related carbon sources, nitrogen source and
The selection of inorganic salts is to fall be familiar with the Specialized Quality and routine technology category of technique personage.
According to the present invention, it is to fall in a scope in 0.05h that this, which continuously ferments,-1To 0.2h-1Dilution rate under and by into
OK.In the preferred embodiment of the present invention, it is in 0.1h that this, which continuously ferments,-1Dilution rate under and carried out.
As used in this article, term " dilution rate (dilution rate) " means feed rate (feed rate) phase
For the ratio of the volume of the fermentation culture medium in fermentation tank.
According to the present invention, this, which continuously ferments, is carried out under an anaerobic condition.
According to the present invention, which can be directly by the clostridium in fermentation tank
The butanol that the bacterial strain of species is generated is removed from fermentation culture medium, extra without fermentation culture medium first is moved to one
Unit.The original position air lift can be known in detail using this those skilled in the art is familiar with and usual technology carries out.In this regard, Ke Yican
Examine, for example, Ezeji T.C.et al. (2003) (with above-mentioned).
Although it is understood that, the operating condition in relation to air lift in situ can further with used carrier gas species with
And the factor such as butanol content and changed, so as to up to causing optimal extraction effect.And the selection of these operating conditions is to be familiar with this
What item those skilled in the art can decide in its sole discretion to routine.
It is preferred that the carrier gas is to be selected from following formed group：Nitrogen, carbon dioxide, hydrogen, and they
Combination.In the preferred embodiment of the present invention, which is nitrogen.
According to the present invention, the original position air lift be a scope fall under the carrier gas flow rate of 1LPM to 4LPM and by into
OK.In the preferred embodiment of the present invention, which carried out under the carrier gas flow rate of 1LPM.At this
In the more preferably concrete example of one of invention, which carried out under the carrier gas flow rate of 2LPM.
<Embodiment>
General experiment material：
1. gas chromatography (gas chromatography, GC) is analyzed：
In the following embodiments, the concentration of butanol contained in each sample to be tested is to use a gas chromatograph
(gas chromatograph, GC) (Hewlett Packard HP 5890Series II) is measured.The related gas phase
The operations condition of chromatograph is shown in table 1 below.
The operating condition of 1. gas chromatograph of table
In addition, for comparing, to be used as correction using the butanol (being purchased from Alfa Aesar) (2-30g/L) of various concentrations
Standard items (control standard) simultaneously carry out identical analysis.
2. dinitrosalicylic acid (dinitrosalicylic acid, DNS) is analyzed：
First, by 3, the 5- dinitrosalicylic acids (3,5-dinitrosalicylic acid) of 1g, the potassium tartrate of 30g
Sodium hydroxide (the sodium of sodium tetrahydrate (potassium sodium tartrate tetrahydrate) and 1.6g
Hydroxide the deionized water of 100mL) is assigned in, and it is spare to obtain a DNS reagents.Then, the sample to be tested of 1mL, Ran Houyu are taken
To add the DNS reagents of 1mL and be uniformly mixed, then the mixture formed is placed in 100 DEG C of water-bath and is kept away
Light action lasts 10 minutes.Afterwards, which is placed in and is cooled down on ice, then with a light splitting under the wavelength of 550nm
Photometer (spectrophotometer) (SpectraMax M3, Molecular Devices) measures light absorption value (OD550)。
By obtained OD550Light absorption value is according to the OD with the glucose with different concentration knowns relative to their own in advance550Inhale
A standard curve made by light value and be converted into glucose content (g/L).
Embodiment 1. is with the clostridium acetobutylicum (Clostridium acetobutylicum) of immobilization and gas in situ
(in situ gas stripping) is carried to continuously generate butanol：
Experiment material：
1. in the present embodiment used reinforced clostridial medium (reinforced clostridial medium,
RCM) it is available from Merck (Cat.No.1.05411.0500).
2. used fermentation medium has a formula as shown in Table 2 below in the present embodiment.
The formula of 2. fermentation medium of table
3. prepare the inoculation source of clostridium acetobutylicum (Clostridium acetobutylicum)
(inoculum)：
First, by purchased from Foodstuff Industrial and Development Inst. of TaiWan, China (Food Industry Research and
Development Institute, FIRDI) living resources preserve and research center (Biosource Collection and
Research Center, BCRC) clostridium acetobutylicum BCRC 10639 (corresponding to ATCC 824) with one for 5% (v/v)
Inoculum concentration is seeded in RCM culture mediums, and heat shock response (heat is carried out under an anaerobic condition and in 80 DEG C of water-bath
Shock reaction) last 2 minutes.Then, after cooling to room temperature, under an anaerobic condition and a constant incubator
Carried out in (37 DEG C, 150rpm) culture last 24 it is small when, in order to activated strains, and thus obtained culture is normally used as
The inoculation source of clostridium acetobutylicum in the present embodiment.
4. prepare fixation support (immobilization carrier)：
First, one is broken into pieces purchased from the clay brick (clay brick) of building materials row using an iron hammer, then in order
Sieve using aperture as 2.38mm and 0.84mm is screened, and obtains that there is a scope to fall in 0.84mm to 2.38mm whereby
The clay brick particle (clay brick particle) of interior particle diameter.Afterwards, the clay brick particle is given with deionized water
Cleaning for several times, is then gone out with sterilization kettle (HVE-50Autoclave, Hirayama) under 121 DEG C and 1.2 atmospheric pressures
Bacterium lasts 30 minutes.Thus obtained sterilized clay brick particle is normally used as the fixation support in the present embodiment.
5. the reactor that continuously ferments (continuous fermentation reactor)：
The structure of the reactor that continuously ferments used in the present embodiment is shown in Fig. 1.The reaction of continuously fermenting
Device goes out comprising a fermentation tank 1 (fermenter 1), the feed inlet 2 (feed port 2) connected respectively with the fermentation tank 1, one
Material mouth 3 (discharge port 3), a gas access 4 (gas inlet 4) and 5 (gas outlet of a gas vent
, and a condensing unit 6 (condensation device 6) for being communicated in the gas vent 5 5).Fresh fermentation medium
It can be fed to a flow velocity fixed via the feed inlet 2 in the fermentation tank 1 for butanol fermentation, and in the fermentation tank 1
In fermentation culture medium can be discharged to identical flow velocity via the discharge port 3 outside the fermentation tank 1, and cause in the fermentation
Fermentation culture medium in groove 1 can be continuously replaced with the speed [that is, dilution rate (dilution rate)] fixed
For fresh fermentation medium, and the content in the fermentation tank 1 can be maintained at the volume of a fixation.For air lift
Carrier gas (carrier gas) can be passed into the fermentation tank 1 with spontaneous with a flow velocity fixed via the gas access 4
Butanol is extracted in ferment culture.The butanol being extracted can enter in the condensing unit 6 via the gas vent 5, and
It is collected by the condensation of the condensing unit 6.
Experimental method：
First, it is the 4th " the preparing fixation support " of the foundation of 420g " experiment material " above is obtained sterilized
The particles filled fermentation tank 1 to the reactor that continuously ferments of clay brick in.Then, by the foundation of 63mL above " experiment material "
The inoculation source of the 3rd " inoculation source for preparing clostridium acetobutylicum " obtained clostridium acetobutylicum is seeded in fermentation tank 1, and
Culture lasts 30 minutes to carry out cell fixation effect under an anaerobic condition and at 37 DEG C.Afterwards, by 2.1L as above
Fermentation medium shown in face table 2 is added into fermentation tank 1, so that the clay brick particle in fermentation tank 1 is with one
The filling rate (filling rate) of 20% (w/v, g/L), then makes the temperature that fermentation tank 1 is 37 DEG C in an anaerobic condition, one
And one for 50rpm stir speed (S.S.) under carry out Batch fermentation.From when proceed by fermentation reaction the 24th is small, by stirring speed
Rate is adjusted to 0rpm.It is 0.1h with one from when proceed by fermentation reaction the 48th is small-1Dilution rate (dilution
Rate) fermentation culture medium is replaced with to fresh fermentation medium, in order to continuously ferment, fermented instead until proceeding by
The 288th after answering terminates when small.
During continuously fermenting, come to carry out the fermentation culture medium in fermentation tank 1 air lift in situ according to following condition, with
Continue to remove butanol from fermentation culture medium：The phase for (calling the stage 1 in the following text) when proceed by fermentation reaction the 48th to 167 is small
Between, do not carry out air lift；During (calling the stage 2 in the following text) when proceed by fermentation reaction the 168th to 215 is small, using with
The nitrogen of the flow velocity (flow rate) of 1LPM carries out air lift；And when proceed by fermentation reaction the 216th to 288 is small
During (calling the stage 3 in the following text), air lift is carried out using the nitrogen of the flow velocity with 2LPM.
During continuously fermenting, sampling is carried out by collecting the fermentation culture medium of 10mL from discharge port 3, in rank
Section 1,2 and 3 in a period of sampling number be respectively 16,7 and 9 times, every about 7 to 8 it is small when sample 1 time.It will take every time
Fermentation culture medium collected by sample gives centrifugation under 13,300rpm and lasts 5 minutes, and obtained supernatant is according to above
Method of 2nd " the dinitrosalicylic acid analysis " of " general experimental method " described in carries out the measurement of glucose content.
It is with Portugal contained in measured fermentation culture medium in relation to glucose utilization rate (glucose utilization rate)
Grape sugared content substitutes into following equation (1) and is calculated：
Formula (1)：A=[(B-C)/B] × 100
Wherein：A=glucose utilizations rate (%)
The glucose content (g/L) of B=fermentation primary fermentation culture mediums
The Portugal for the fermentation culture medium that each sampling time points of C=measure respectively
Grape sugared content (g/L)
Then, the average glucose utilization rate (%) in each stage is further calculated.Obtained experimental data be with
Average value ± standard deviation (Standard Deviation, S.D.) represents.
In addition, obtained supernatant above (is purchased from Merck by 0.2 μm of syringe-driven filter of aperture
Millipore) filtered, and obtained filtrate is the 1st " gas chromatography point according to " general experimental method " above
Method of the analysis " described in carries out the content analysis of butanol.Related butanol yield (butanol productivity) be with
Contained butanol content substitutes into following equation (2) and is calculated in measured fermentation culture medium：
Formula (2)：D=E × F
Wherein：D=butanol yield (g/Lh)
The fourth for the fermentation culture medium that each sampling time points of E=measure respectively
Alcohol content (g/L)
F=dilution rates (h-1)
Then, the average butanol yield (g/Lh) in each stage is further calculated.Obtained experimental data is with flat
Mean value ± standard deviation represents.
In addition, respectively when last the 12 of the stage 2 is small (that is, when small in proceed by fermentation reaction the 204th to 215)
And the stage 3 last 12 it is small when (that is, when proceed by fermentation reaction the 277th to 288 is small) during, use condensation
Device 6 collects condensate liquid, and the condensate liquid of 10mL is collected after the volume for noting down the condensate liquid collected by 12 hours.Separately
Outside, the fermentation culture medium of 10mL is collected from fermentation tank 1 at the end of stage 2 and stage 3 respectively.Then, by condensate liquid and
Fermentation culture medium lasts 5 minutes respectively at giving centrifugation under 13,300rpm, then by obtained supernatant using aperture as 0.2 μ
The syringe-driven filter of m is filtered.And obtained filtrate is the 1st " the gas phase layer according to " general experimental method " above
Method of the analysis analysis " described in carries out the content analysis of butanol.Related butanol removes speed (butanol removal
Rate it is) so that butanol content contained in measured condensate liquid substitutes into following equation (3) and is calculated：
Formula (3)：G=(H × I)/J
Wherein：G=butanol removes speed (g/h)
The butanol content (g/L) of H=condensate liquids
The volume (L) of I=condensate liquids
J=12 (h)
And it is by measured condensate liquid and fermentation culture medium in relation to butanol selection rate (butanol selectivity)
Contained butanol content is converted into percentage by weight and substitutes into following equation (4) and be calculated：
Formula (4)：K=[L/ (1-L)]/[M/ (1-M)]
Wherein：K=butanol selection rates
Butanol percentage by weight (wt%) in L=condensate liquids
Butanol percentage by weight (wt%) in M=fermentation culture mediums
Experimental result：
The measured result of this experiment is shown in table 3 below.
The average glucose utilization rate in 3. each stage of table, average butanol yield, butanol remove speed and butanol selection
Rate
As seen from Table 3, all it is significantly higher than rank in the average glucose utilization rate in stage 2 and 3 and average butanol yield
1 tool person of section.This experimental result is shown：During continuously fermenting, using nitrogen come carry out air lift in situ can be notable
Ground lifts the utilization of glucose, and then lifts butanol yield.
In addition, the butanol in stage 3, which removes speed and butanol selection rate, is all considerably higher than 2 tool persons of stage.This reality
Test the results show：To carry out air lift in situ with a preferable Butanol Recycling effectiveness with one for the nitrogen of the flow velocity of 2LPM.
All patents for being quoted in this specification and document are merged in this case as referring to data using its entirety.If have
When conflicted, this case describes in detail will get the upper hand (comprising in being defined in).
Although the present invention is described with reference to above-mentioned specific concrete example, it will be apparent that without departing substantially from the scope of the present invention and essence
God is lower can to make many modifications and variations.Therefore it is intended that the present invention is only by such as with the attached claim institute of text inspection
The limitation for the person of showing.
Claims (8)
- A kind of 1. method for producing butanol, it is characterised in that：This method includes：The bacterial strain of one Clostridial species is fixed on a fixation support；AndMake the bacterial strain through fixed Clostridial species continuously ferment, and make it that butanol is generated, wherein, butanol is even It is recovered during supervention ferment by using the air lift in situ of a carrier gas.
- 2. according to the method described in claim 1, it is characterized in that：The fixation support is to be selected from following formed group Group：Clay brick, zeolite, ceramics, bone black, resin, sawdust, coke, rice husk, cotton, cornstalk, calcium alginate, silica gel, and they Combination.
- 3. according to the method described in claim 1, it is characterized in that：The carrier filling rate of the fixation support is fallen 20% (w/v, g/L) is in the range of 40% (w/v, g/L).
- 4. according to the method described in claim 1, it is characterized in that：The carrier gas is to be selected from following formed group： Nitrogen, carbon dioxide, hydrogen, and combinations thereof.
- 5. according to the method described in claim 1, it is characterized in that：The original position air lift is to fall in a scope in 1LPM to 4LPM Carried out under carrier gas flow rate.
- 6. according to the method described in claim 1, it is characterized in that：The bacterial strain of the Clostridial species is formed selected from following Group：Clostridium acetobutylicum (Clostridium acetobutylicum), clostridium saccharoacetoperbutylicum (Clostridium Saccharoperbutylacetonicum), Clostridium beijerinckii (Clostridium beijerinckii), clostridium tyrobutyricum (Clostridium tyrobutyricum), clostridium butyricum (Clostridium butyricum), and combinations thereof.
- 7. according to the method described in claim 1, it is characterized in that：It is to fall in a scope in 0.05h that this, which continuously ferments,-1Extremely 0.2h-1Dilution rate under and carried out
- 8. according to the method described in claim 1, it is characterized in that：This, which continuously ferments, is carried out under an anaerobic condition.
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| CN112955559A (en) * | 2018-09-28 | 2021-06-11 | Ifp 新能源公司 | Method for producing alcohol using clostridium on solid carrier |
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