CN105713823A - Device and method for producing isopropanol and butanol through extraction and fermentation - Google Patents
Device and method for producing isopropanol and butanol through extraction and fermentation Download PDFInfo
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M21/00—Bioreactors or fermenters specially adapted for specific uses
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
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- 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
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- 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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Abstract
The invention discloses a device and method for producing isopropanol and butanol through extraction and fermentation. The device comprises a common bioreactor, a fiber bed immobilized reactor and a liquid-liquid extractor which are serially connected through a pipeline. The fiber bed immobilized reactor is connected with the common bioreactor through a reflux pipeline to form a first liquid circulation system. The extractor is connected with the bioreactor through a reflux pipeline to form a second liquid circulation system. Firstly, a seed solution of ethanol producing strains is cultured in the bioreactor, the strains are fixed to the fiber bed immobilized reactor when growing to a logarithmic growth metaphase and are fermented to an ethanol producing stage, and fermented liquid is led to the extractor for extraction and finally flows back to the bioreactor. An extraction solvent is added to the extractor and is lauric acid dissolved in an organic medium, and the organic medium is non-water-soluble liquid. The device and the method can effectively improve the substrate conversion rate and improve the ethanol producing amount and ethanol producing ratio of strains.
Description
Technical field
The invention belongs to bioengineering field, relate to isopropanol and the apparatus and method of butanol, before immunoassay fermentation, in the extractive fermentation system that a fibre bed immobilization reactor and liquid-liquid extractor are connected, carry out isopropanol and the apparatus and method of butanol on-line continuous extractive fermentation in particular with lauric acid/dodecanoic acid.
Background technology
Isopropanol and butanol be two kinds at the very important organic solvent of chemical field, it is possible to be used for doing the basic framework of solubilising reagent and chemosynthesis.What is more important is high due to they energy density, can dissolve each other with arbitrary proportion with gasoline and diesel oil, and biotechnology can be used to carry out fermenting and producing, so being considered as ideal one of recyclable fuel or fuel additive.(refer to non-patent literature: 1. Palsson, B., Fathi-Afshar, S., Rudd, D., Lightfoot, E.1981.Biomassasasourceofchemicalfeedstocks:aneconomicev aluation.Science, 213 (4507), 513-517；2. Lee, S.Y., Park, J.H., Jang, S.H., Nielsen, L.K., Kim, J., Jung, K.S.2008.FermentativebutanolproductionbyClostridia.Biote chnologyandbioengineering, 101 (2), 209-228).
But, its murder by poisoning producing bacterial strain is made yield always all very low by the fermentation of isopropanol and butanol, and the high boiling point of butanol makes to separate it from fermentation liquid and becomes to be difficult to very much, and extractive fermentation can alleviate this difficult problem effectively.Extractive fermentation is a fermentation method for producing that can effectively reduce Product inhibiton, increase substrate conversion efficiency and alcohols output capacity, and it is while reducing Product inhibiton promotion product output, it is achieved that efficiently separating and purifying purpose product.Liquid-liquid extraction is that one utilizes fluid extractant to process immiscible bi-component or Multi component with it, it is achieved the mass transfer separation process of Component seperation.Due to it have normal-temperature operation, save the energy, simple to operate, be not related to the advantage such as solid, gas, be therefore widely used in multiple fields such as chemistry, chemical industry and biology.A lot of researchs report employing vegetable oil, fatty acid ester, alkane and long-chain fatty alcohol and butanol carry out liquid-liquid situ extracting fermentation and (refers to non-patent literature: OffemanRD, StephensonSK, RobertsonGH, OrtsWJ.2006.Solventextractionofethanolfromaqueoussolutio nsusingbiobasedoils, alcohols, andesters.JAmOilChemSoc83 (2): 153 156).But these abstraction techniques are affected by the restriction of the limited solvent partition coefficient of extractant and fail effectively to improve solvent extraction effect always.Therefore, seek a kind of to have compared with high fat alcohol partition coefficient, nontoxic, water insoluble and there is the extractant of higher biological interoperability for using liquid-liquid situ extracting fermentation technique to produce isopropanol and butanol is particularly important.
Summary of the invention
One of the object of the invention is to use the lauric acid/dodecanoic acid being dissolved in organic media to carry out liquid-liquid original position on-line continuous extractive fermentation to produce isopropanol and butanol, while fermentation, two kinds of alcohol can be separated effectively from culture medium and be purified, reduce product inhibition in culture medium.
Extractor is together in series with fibre bed immobilization reactor and bioreactor by the two of the object of the invention by pipeline, then carries out isopropanol and butanol, before immunoassay fermentation together with extractant.
A kind of extractive fermentation produces the device of isopropanol and butanol, is sequentially connected in series common bioreactor, fibre bed immobilization reactor and liquid-liquid extractor including by pipeline；Described fibre bed immobilization reactor is connected by reflux line with bioreactor again, constitutes first fluid circulation system；Described extractor is connected by reflux line with bioreactor again, constitutes second fluid circulation system；This device can control liquid from bioreactor, through different fluid circulation systems, finally returns again to bioreactor.
Injection port and outlet it is respectively provided with at extractor 1/4 height and position and bottom.
A kind of said apparatus on-line continuous extractive fermentation produces the method for isopropanol and butanol, in bioreactor, first cultivate the seed liquor producing alcohol bacterium, treat that it is to mid log phase, strain is fixed in fibre bed immobilization reactor, fermentation is to entering the product alcohol stage, fermentation liquid is passed into extractor extract, be finally back to bioreactor again；Added with extractant in described extractor, extractant is the dodecylic acid being dissolved in organic media, and described organic media is non-water-soluble liquid.
Described extractant is dissolved in organic media by 2/5～3/5 (W/W).It is highly preferred that described extractant is dissolved in organic media by 1/2 (W/W).
Described organic media includes oleyl alcohol, Oleum Brassicae campestris, Petiolus Trachycarpi oil or Jatropha curcas oil.
When being fixed of strain, open first fluid circulation system, make culture medium only circulate between bioreactor and fibre bed immobilization device；When fermentation liquid cell concentration no longer declines and starts to continue to raise, change fresh fermentation medium and complete cell fixation.
When fermentation enters and produces the alcohol stage, close first fluid circulation system, make fermentation liquid flow in extractor and reach extractor 1/4 At The Height (organic extractant phase agent is on upper strata, aqueous phase fermentation liquid is in lower floor), then start stirring at low speed to extract, and open second fluid circulation system, allow culture medium flow back to bioreactor.
Mixing speed during extraction controls at 100～150rpm, makes fermentation liquid can have with extractant under the stirring at low speed of extractor and is significantly separated, and controls to make fermentation liquid leave when extractor is back to bioreactor by mixing speed and does not carry extractant.
When OD continuous decrease in bioreactor, terminate extractive fermentation, change fresh fermentation broth, carry out the extractive fermentation of a new round；Continuously repeat above-mentioned extractive fermentation, until extractant loses effect of extracting；Terminate extractive fermentation, complete once complete isopropanol and butanol on-line continuous extractive fermentation process.
Whole device is first sterilizing before using, and passes into aseptic high pure nitrogen wherein so that it is become the environment of a strictly anaerobic.
Product alcohol bacterial strain of the present invention is the anaerobism clostridium producing isopropanol and butanol or all product butanol, it does not have limit especially.Can enumerate bacterial strain is Clostridium beijerinckii (Clostridiumbeijerinckii), clostridium acetobutylicum (Clostridiumacetobutylicum), sugar acetic acid Clostridium acetobutylicum (Clostridiumsaccharoperbutylacetonicum) and clostridium saccharobutyricum (Clostridiumsaccharobutylicum) four kinds, and the present invention preferably produces the Clostridium beijerinckii of isopropanol and butanol.
The fermentation temperature of the present invention is 36 DEG C；Medium's PH Value is 6.5, and sweat does not regulate and control pH；Fermentation volume is 2.5L；Extraction temperature is 30～36 DEG C；Fermentation liquid circulation flow velocity is 50ml/min.
The present invention has the advantages that and advantage:
1. long-chain fatty acid dodecylic acid (also known as lauric acid) is applied to the extractive fermentation of isopropanol and butanol by the present invention, it is intended to improve the effect of liquid-liquid situ extracting fermentation, it is achieved to isopropanol and butanol efficiently extract and separate and fermentation online.Lauric acid/dodecanoic acid has water-insoluble, produces alcohol unrestraint effect to strain growth nonhazardous with to cell, and has higher butanol partition coefficient, it is possible to the isopropanol of low concentration and butanol in active adsorption fermentation liquid.
2. the lauric acid/dodecanoic acid being dissolved in organic media can reduce the concentration of alcohol in fermentation liquid quickly and efficiently, this is the difference to alcohols partition coefficient due to fermentation liquid and extractant, alcohols priority allocation in fermentation liquid is in extractant, and reaches higher concentration wherein, thus reducing product inhibition.
3. utilize lauric acid/dodecanoic acid situ extracting and fermentation coupling, it is possible to improve the utilization rate of glucose, be effectively improved substrate conversion efficiency, improve the production intensity of fermentation, improve the product alcohol amount of bacterial strain and produce alcohol rate.
4. utilize lauric acid/dodecanoic acid original position continuous extraction to ferment, it is possible to be enriched in extractant by alcohols with higher concentration, be effectively reduced later separation purification difficulty and cost.
Accompanying drawing explanation
Fig. 1 is the structural representation that extractive fermentation of the present invention produces the device of isopropanol and butanol.
Fig. 2 is output and the concentration change situation of four continuous extractions fermentation isopropanol and butanol.
Detailed description of the invention
Below in conjunction with accompanying drawing, the present invention will be further described.
The product alcohol bacterial strain used in all embodiments of the invention is Clostridium beijerinckii (Clostridiumbeijerinckii, ATCC6014).
The seed culture medium (CM149) (g/L) used in all embodiments of the invention: yeast extract 3g, beef extract powder 10g, peptone 10g, glucose 5g, soluble starch 1g, NaCl5g, sodium acetate 3g, cysteine hydrochloride 0.5g, pH6.8 ± 0.2.
The fermentation medium (P2) (g/L) (ml/L) used in all embodiments of the invention: glucose 50g, yeast extract 3g, acetate buffer mother solution 10ml, vitamin solution mother solution 10ml, mineral salt solution 10ml, original ph is 6.5.
The mother solution formula of above-mentioned each solution following (solution is filtration sterilization):
1. acetate buffer mother solution (g/L): KH2PO450g, K2HPO450g, ammonium acetate 220g；
2. vitamin solution mother solution (g/L): para-amino benzoic acid 0.1g, biotin 0.001g, thiamine 0.1g；
3. mineral salt solution (g/L): MgSO420g, MnSO41g, FeSO4·7H2O1g, NaCl1g.
The invention provides the apparatus and method using the lauric acid/dodecanoic acid being dissolved in organic media to carry out liquid-liquid original position on-line continuous extractive fermentation production isopropanol and butanol, comprise the steps:
1. the assembling of extractive fermentation device and sterilizing, as shown in Figure 1: include feed supplement bottle 1, bioreactor 2, peristaltic pump 3, fibre bed immobilization reactor 4, liquid-liquid extractor 5 and check valve (11).It is sequentially connected in series common bioreactor 2, fibre bed immobilization reactor 4 and liquid-liquid extractor 5 by pipeline；Described fibre bed immobilization reactor 4 is connected by the first reflux line 6 with bioreactor 2 again, constitutes first fluid circulation system；Described extractor 5 is connected by the second reflux line 7 with bioreactor 2 again, constitutes second fluid circulation system.
2. first by involved apparatus sterilizing, then fibre bed immobilization reactor and liquid-liquid extractor are coupled together by pipeline, then connect into a flowable blood circulation of liquid with bioreactor.With pure nitrogen gas, the air displacement in extractive fermentation device is fallen so that it is become the environment of an abundant anaerobism.
3. with pure nitrogen gas, the air displacement in extractive fermentation device is fallen so that it is become the environment of an abundant anaerobism.
4. the preparation of extractant: extractant is dissolved in water-insoluble non-toxic aq organic media in 1/2 (W/W) ratio, is subsequently poured into extractor.
5. spawn culture and immobilization: first cultivate in bioreactor and produce alcohol bacterium seed liquor, treat bacterial concentration OD600When reaching more than 2.0, open peristaltic pump and fermentation liquid is circulated in systems, thus cell is fixed on fibre bed immobilization reactor.When fermentation liquid cell concentration no longer declines and starts to continue to raise, change fresh fermentation medium and complete bacterial strain immobilization.In this process, culture medium circulation is not passed through extractor and walks the first reflux line 6.
6., after changing fresh culture, when fermentation is in the initial stage producing acid phase, culture medium circulation is not passed through extractor and walks the first reflux line 6.
7. the alcohol stage (about fermentation initial after 12 hours) is produced when fermentation enters, close the valve on the first reflux line 6, make culture medium flow in extractor 5 and reach about 1/4 extractor volume (organic extractant phase agent is on upper strata, aqueous phase culture medium is in lower floor), then start stirring at low speed to extract, and open and allow culture medium flow back to bioreactor 2.This process to control mixing speed, make it can also have with extractant under the stirring at low speed of extractor to be significantly separated, ensure that fermentation liquid leaves not carry when extractor flows back to bioreactor and mix extractant, in order to avoid causing extractant with fermentation liquid circulation in extractive fermentation system.
8. when fermentation liquid to the inhibitory action of strain very strong time, terminate extractive fermentation, change fresh fermentation broth, enter and produce alcohol after date and continue to carry out new one batch of fermentation, extraction with identical extractant.
9. step extractive fermentation 7. is continuously repeated, until extractant loses effect of extracting；
10. terminate extractive fermentation, complete once the complete process utilizing lauric acid/dodecanoic acid to carry out liquid-liquid original position on-line continuous extractive fermentation production isopropanol and butanol.
The preparation of extractant and preparation: lauric acid/dodecanoic acid is dissolved in water-insoluble non-toxic aq organic media in the ratio of 1/2 (W/W), is subsequently poured into extractor.The organic media that the present embodiment adopts is oleyl alcohol.
Embodiment 1: the extractant of the equivalent fermentating liquid volume of use carries out first of extractive fermentation continuously
1. spawn culture and immobilization: first cultivate in bioreactor 2 and produce alcohol bacterium seed liquor, treat bacterial concentration (OD600) when reaching more than 2.0, open peristaltic pump 3 and fermentation liquid is circulated in first fluid circulation system, thus cell is fixed on fibre bed immobilization reactor.When fermentation liquid cell concentration no longer declines and starts to continue to raise, change fresh fermentation medium and complete bacterial strain immobilization.In this process, culture medium circulation is not passed through extractor 5.After changing fresh culture, when fermentation is in the initial stage producing acid phase, culture medium circulation is not passed through extractor.
2. liquid-liquid situ extracting fermentation: produce the alcohol stage (about fermentation initial after 12 hours) when fermentation enters, close the valve on the first reflux line 6, make culture medium flow in extractor 5 and reach about 1/4 extractor volume (organic extractant phase agent is on upper strata, aqueous phase culture medium is in lower floor), then start stirring at low speed to extract, and the valve opened on the second reflux line 7 flows back to bioreactor 2 by culture medium.This process to control mixing speed at 100～150rpm, make it can have with extractant under the stirring at low speed of extractor to be significantly separated, ensure that fermentation liquid leaves when extractor flows back to bioreactor and do not carry assorted extractant, in order to avoid causing extractant with fermentation liquid circulation in extractive fermentation system.When fermentation liquid to the inhibitory action of strain very strong time, terminate extractive fermentation.
3. through the concrete data statistics of the extractive fermentation of 90 hours, isopropanol and production of butanol such as table 1.Utilize lauric acid/dodecanoic acid to carry out the fermentation of liquid-liquid situ extracting and can be effectively improved the yield of isopropanol and butanol, and effectively both are extracted in organic facies.
4. relative to common non-extractive fermentation, isopropanol and the butanol unit volume productivity of this batch of extractive fermentation have been respectively increased 92.90% and 83.38%, and total alcohol unit volume productivity improves 89.91%.
5. relative to common non-extractive fermentation, this batch of total alcohol conversion improves 5.13%；Total alcohol unit volume productivity improves 32.0%.
Table 1: produce isopropanol and butanol for carbon source extractive fermentation with glucose: first of continuous extraction fermentation
Embodiment 2: use the extractant of equivalent fermentating liquid volume to carry out the second batch of extractive fermentation continuously
1. the bacterial strain that this embodiment is to continue with using embodiment 1 fixing produces, and uses embodiment 1 to extract the extractant after using simultaneously and proceeds to extract.So the fixing replacing with extractant of strain need not be carried out.
2. liquid-liquid situ extracting fermentation: produce the alcohol stage (about fermentation initial after 12 hours) when fermentation enters, close the valve on the first reflux line 6, make culture medium flow in extractor 5 and reach about 1/4 extractor volume (organic extractant phase agent is on upper strata, aqueous phase culture medium is in lower floor), then start stirring at low speed to extract, and the valve opened on the second reflux line 7 flows back to bioreactor 2 by culture medium.This process to control mixing speed at 100～150rpm, make it can have with extractant under the stirring at low speed of extractor to be significantly separated, ensure that fermentation liquid leaves when extractor flows back to bioreactor and do not carry extractant, in order to avoid causing extractant with fermentation liquid circulation in extractive fermentation system.When fermentation liquid to the inhibitory action of strain very strong time, terminate extractive fermentation.
3. through the concrete data statistics of the extractive fermentation of 90 hours, isopropanol and production of butanol such as table 2.Utilize the used lauric acid/dodecanoic acid of embodiment 1 to carry out liquid-liquid situ extracting fermentation and can also effectively increase the yield of isopropanol and butanol equally, and effectively both are extracted in organic facies.
4. relative to common non-extractive fermentation, isopropanol and the butanol unit volume productivity of this batch of extractive fermentation have been respectively increased 104.11% and 92.1%, and total alcohol unit volume productivity improves 95.52%.
5. relative to common non-extractive fermentation, this batch of total alcohol conversion improves 10.26%；Total alcohol unit volume productivity improves 36.0%.
Table 2: produce isopropanol and butanol with glucose for carbon source extractive fermentation: the second batch of continuous extraction fermentation
Embodiment 3: the extractant of the equivalent fermentating liquid volume of use carries out the 3rd batch of extractive fermentation continuously
1. this embodiment is that the bacterial strain being continuing with embodiment 1 after embodiment 2 fixing produces, and the extractant after simultaneously using embodiment 1 and embodiment 2 continuous extraction to use proceeds to extract.So the fixing replacing with extractant of strain need not be carried out.
2. liquid-liquid situ extracting fermentation: produce the alcohol stage (about fermentation initial after 12 hours) when fermentation enters, close the valve on the first reflux line 6, make culture medium flow in extractor and reach about 1/4 extractor volume (organic extractant phase agent is on upper strata, aqueous phase culture medium is in lower floor), then start stirring at low speed to extract, and the valve opened on the second reflux line 7 flows back to bioreactor by culture medium.This process to control mixing speed at 100～150rpm, make it can have with extractant under the stirring at low speed of extractor to be significantly separated, ensure that fermentation liquid leaves when extractor flows back to bioreactor and do not carry extractant, in order to avoid causing extractant with fermentation liquid circulation in extractive fermentation system.When fermentation liquid to the inhibitory action of strain very strong time, terminate extractive fermentation.
3. through the concrete data statistics of the extractive fermentation of 90 hours, isopropanol and production of butanol such as table 3.Utilize embodiment 1 and the continuous used lauric acid/dodecanoic acid of embodiment 2 to carry out liquid-liquid situ extracting fermentation and can also effectively increase the quantum of output of isopropanol and butanol equally, and effectively both are extracted in organic facies.
4. relative to common non-extractive fermentation, isopropanol and the butanol unit volume productivity of this batch of extractive fermentation have been respectively increased 88.97% and 46.29%, and total alcohol unit volume productivity improves 60.69%.
5. relative to common non-extractive fermentation, this batch of total alcohol conversion has a declining tendency, and decreases 2.56%；Total alcohol unit volume productivity improves 12.0%.
Table 3: produce isopropanol and butanol with glucose for carbon source extractive fermentation: the 3rd batch of continuous extraction fermentation
Embodiment 4: the extractant of the equivalent fermentating liquid volume of use carries out the 4th batch of extractive fermentation continuously
1. this embodiment is that the bacterial strain being continuing with embodiment 1 after first three embodiment fixing produces, and the extractant after simultaneously using first three embodiment continuous extraction to use proceeds to extract.So the fixing replacing with extractant of strain need not be carried out.
2. liquid-liquid situ extracting fermentation: produce the alcohol stage (about fermentation initial after 10 hours) when fermentation enters, close the valve on the first reflux line 6, make culture medium flow in extractor and reach about 1/4 extractor volume (organic extractant phase agent is on upper strata, aqueous phase culture medium is in lower floor), then start stirring at low speed to extract, and the valve opened on the second reflux line 7 flows back to bioreactor by culture medium.This process to control mixing speed at 100～150rpm, make it can also have with extractant under the stirring at low speed of extractor to be significantly separated, ensure that fermentation liquid leaves when extractor flows back to bioreactor and do not carry extractant, in order to avoid causing extractant with fermentation liquid circulation in extractive fermentation system.When fermentation liquid to the inhibitory action of strain very strong time, terminate extractive fermentation.
3. through the concrete data statistics of the extractive fermentation of 80 hours, isopropanol and production of butanol such as table 4.Utilize first three time continuous used lauric acid/dodecanoic acid of embodiment to carry out liquid-liquid situ extracting fermentation and can also improve the yield of isopropanol and butanol equally, and a small amount of alcohols is extracted in organic facies, but effect of extracting effect of extracting several times before can not show a candle to.
4. relative to common non-extractive fermentation, isopropanol and the butanol unit volume productivity of this batch of extractive fermentation have been respectively increased 65.23% and 33.52%, and total alcohol unit volume productivity improves 44.23%.
5. relative to common non-extractive fermentation, this batch of total alcohol conversion is not changed in；Total alcohol unit volume productivity improves 16.0%.
6. four extractive fermentations, the isopropanol being enriched with in extractant after continuous extraction is 24.4g, and butanol is 47.23g, and the total alcohol amount being enriched with is 71.63g.
7., in four continuous extractions are fermented, the output of isopropanol and butanol and concentration change situation are as shown in Figure 2.
Table 4: produce isopropanol and butanol with glucose for carbon source extractive fermentation: the 4th batch of continuous extraction fermentation
Claims (10)
1. the device of an extractive fermentation production isopropanol and butanol, it is characterised in that include being sequentially connected in series common bioreactor, fibre bed immobilization reactor and liquid-liquid extractor by pipeline；Described fibre bed immobilization reactor is connected by reflux line with bioreactor again, constitutes first fluid circulation system；Described extractor is connected by reflux line with bioreactor again, constitutes second fluid circulation system.
2. device according to claim 1, it is characterised in that: injection port and outlet it is respectively provided with at extractor 1/4 height and position and bottom.
3. one kind utilizes the method that device on-line continuous extractive fermentation described in claim 1 or 2 produces isopropanol and butanol, it is characterized in that, in bioreactor, first cultivate the seed liquor producing alcohol bacterium, treat that it is to mid log phase, strain is fixed in fibre bed immobilization reactor, fermentation liquid, to entering the product alcohol stage, is passed into extractor and extracts, be finally back to bioreactor again by fermentation；Added with extractant in described extractor, extractant is the dodecylic acid being dissolved in organic media, and described organic media is non-water-soluble liquid.
4. method according to claim 3, it is characterised in that described extractant is dissolved in organic media by 2/5～3/5 (W/W), and described organic media includes oleyl alcohol, Oleum Brassicae campestris, Petiolus Trachycarpi oil or Jatropha curcas oil.
5. method according to claim 4, it is characterised in that described extractant is dissolved in organic media by 1/2 (W/W).
6. method according to claim 5, it is characterised in that during being fixed of strain, open first fluid circulation system, makes culture medium only circulate between bioreactor and fibre bed immobilization reactor；When fermentation liquid cell concentration no longer declines and starts to continue to raise, change fresh fermentation medium and complete cell fixation.
7. method according to claim 3 or 4 or 5 or 6, it is characterized in that, when fermentation enters and produces the alcohol stage, close first fluid circulation system, fermentation liquid is made to flow in extractor and reach extractor 1/4 At The Height, then start stirring at low speed to extract, and open second fluid circulation system, allow culture medium flow back to bioreactor.
8. method according to claim 7, it is characterised in that mixing speed during extraction controls at 100～150rpm.
9. method according to claim 7, it is characterised in that when OD continuous decrease in bioreactor, terminates extractive fermentation, changes fresh fermentation broth, carries out the extractive fermentation of a new round；Continuously repeat above-mentioned extractive fermentation, until extractant loses effect of extracting；Terminate extractive fermentation, complete once complete isopropanol and butanol on-line continuous extractive fermentation process.
10. method according to claim 7, it is characterised in that whole device is first sterilizing before using, and passes into aseptic high pure nitrogen wherein so that it is become the environment of a strictly anaerobic.
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| CN112358986A (en) * | 2020-11-09 | 2021-02-12 | 华南理工大学 | Clostridium butyricum and application thereof in immobilized fermentation production of 1, 3-propylene glycol |
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