WO2017214873A1 - 一种发酵耦合分离纯化生产丁醇的方法及装置 - Google Patents

一种发酵耦合分离纯化生产丁醇的方法及装置 Download PDF

Info

Publication number
WO2017214873A1
WO2017214873A1 PCT/CN2016/085818 CN2016085818W WO2017214873A1 WO 2017214873 A1 WO2017214873 A1 WO 2017214873A1 CN 2016085818 W CN2016085818 W CN 2016085818W WO 2017214873 A1 WO2017214873 A1 WO 2017214873A1
Authority
WO
WIPO (PCT)
Prior art keywords
butanol
acetone
membrane
ethanol
fermentation
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/CN2016/085818
Other languages
English (en)
French (fr)
Inventor
薛闯
朱超
陈丽杰
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Dalian University of Technology
Original Assignee
Dalian University of Technology
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Dalian University of Technology filed Critical Dalian University of Technology
Priority to US15/737,796 priority Critical patent/US11118198B2/en
Priority to PCT/CN2016/085818 priority patent/WO2017214873A1/zh
Publication of WO2017214873A1 publication Critical patent/WO2017214873A1/zh
Anticipated expiration legal-status Critical
Priority to US17/394,695 priority patent/US20210363553A1/en
Ceased legal-status Critical Current

Links

Images

Classifications

    • 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
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D3/00—Distillation or related exchange processes in which liquids are contacted with gaseous media, e.g. stripping
    • B01D3/001—Processes specially adapted for distillation or rectification of fermented solutions
    • B01D3/002—Processes specially adapted for distillation or rectification of fermented solutions by continuous methods
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D3/00—Distillation or related exchange processes in which liquids are contacted with gaseous media, e.g. stripping
    • B01D3/14—Fractional distillation or use of a fractionation or rectification column
    • B01D3/143—Fractional distillation or use of a fractionation or rectification column by two or more of a fractionation, separation or rectification step
    • B01D3/145—One step being separation by permeation
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D5/00—Condensation of vapours; Recovering volatile solvents by condensation
    • B01D5/0033—Other features
    • B01D5/0045—Vacuum condensation
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D61/00—Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis or ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor
    • B01D61/36—Pervaporation; Membrane distillation; Liquid permeation
    • B01D61/362—Pervaporation
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D71/00—Semi-permeable membranes for separation processes or apparatus characterised by the material; Manufacturing processes specially adapted therefor
    • B01D71/02—Inorganic material
    • B01D71/028—Molecular sieves
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D71/00—Semi-permeable membranes for separation processes or apparatus characterised by the material; Manufacturing processes specially adapted therefor
    • B01D71/02—Inorganic material
    • B01D71/028—Molecular sieves
    • B01D71/0281—Zeolites
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D71/00—Semi-permeable membranes for separation processes or apparatus characterised by the material; Manufacturing processes specially adapted therefor
    • B01D71/06—Organic material
    • B01D71/70—Polymers having silicon in the main chain, with or without sulfur, nitrogen, oxygen or carbon only
    • B01D71/701—Polydimethylsiloxane
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C29/00—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom not belonging to a six-membered aromatic ring
    • C07C29/74—Separation; Purification; Use of additives, e.g. for stabilisation
    • C07C29/76—Separation; Purification; Use of additives, e.g. for stabilisation by physical treatment
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C45/00—Preparation of compounds having >C = O groups bound only to carbon or hydrogen atoms; Preparation of chelates of such compounds
    • C07C45/78—Separation; Purification; Stabilisation; Use of additives
    • C07C45/786—Separation; Purification; Stabilisation; Use of additives by membrane separation process, e.g. pervaporation, perstraction, reverse osmosis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C45/00—Preparation of compounds having >C = O groups bound only to carbon or hydrogen atoms; Preparation of chelates of such compounds
    • C07C45/78—Separation; Purification; Stabilisation; Use of additives
    • C07C45/81—Separation; Purification; Stabilisation; Use of additives by change in the physical state, e.g. crystallisation
    • 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
    • C12P7/06—Ethanol, i.e. non-beverage
    • C12P7/065—Ethanol, i.e. non-beverage with microorganisms other than yeasts
    • 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
    • C12P7/24—Preparation of oxygen-containing organic compounds containing a carbonyl group
    • C12P7/26—Ketones
    • C12P7/28—Acetone-containing products
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D2257/00—Components to be removed
    • B01D2257/70—Organic compounds not provided for in groups B01D2257/00 - B01D2257/602
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D2311/00—Details relating to membrane separation process operations and control
    • B01D2311/25—Recirculation, recycling or bypass, e.g. recirculation of concentrate into the feed
    • B01D2311/252—Recirculation of concentrate
    • B01D2311/2523—Recirculation of concentrate to feed side
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D2311/00—Details relating to membrane separation process operations and control
    • B01D2311/26—Further operations combined with membrane separation processes
    • B01D2311/2674—Condensation
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D2311/00—Details relating to membrane separation process operations and control
    • B01D2311/26—Further operations combined with membrane separation processes
    • B01D2311/2688—Biological processes
    • 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
    • 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
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/10—Process efficiency

Definitions

  • the invention relates to a method for separating, purifying and producing butanol, acetone and ethanol (or butanol) by fermentation coupling, and belongs to the field of biotechnology.
  • Butanol and acetone are potential liquid energy sources and important chemicals used in the pharmaceutical and food industries. They can be obtained by microbial fermentation. See Dürre, P. Biobutanol: an attractive biofuel. Biotechnol. J. 2: 1525 –1534, 2007. However, when butanol is fermented by Clostridium acetobutylicum or Clostridium perfringens, the concentration of butanol in the fermentation broth usually does not exceed 2.0% (w/v). Further, the boiling point of butanol is 117.7 ° C, which is higher than the boiling point of water by 100 ° C.
  • the present invention utilizes a vaporization permeation method to couple fermentatively to produce butanol, acetone and ethanol (or butanol), and to separate and purify butanol, acetone and ethanol (or butanol) during the fermentation process. .
  • the invention specifically relates to the following:
  • a method for the separation and purification of butanol, acetone and ethanol or purified butanol by fermentation comprising the steps of:
  • the vaporization infiltration method includes the following steps:
  • the source of the gas bubbles is a self-produced gas or an exogenous gas of a acetone butanol producing bacteria or a butanol producing bacteria
  • the self-produced gas is preferably carbon dioxide and/or hydrogen.
  • the exogenous gas is preferably nitrogen.
  • acetone butanol-producing bacterium is preferably selected from the group consisting of Clostridium acetobutylicum, Clostridium beijerinckii, Escherichia coli or Clostridium tyrobutyricum.
  • the genetically engineered bacteria of these strains preferably Clostridium acetobutylicum, which is a genetically engineered strain producing butanol, preferably producing butanol E. coli or Clostridium butyrate.
  • the membrane is a vaporized permeable membrane, preferably a butoxide and acetone highly selective organic hydrophobic separation membrane or an organic-inorganic composite membrane, more preferably selected from the group consisting of silicone rubber and polytrimethylsilylpropyne.
  • a vaporized permeable membrane preferably a butoxide and acetone highly selective organic hydrophobic separation membrane or an organic-inorganic composite membrane, more preferably selected from the group consisting of silicone rubber and polytrimethylsilylpropyne.
  • a device for fermentative coupling separation and purification of butanol, acetone and ethanol or purified butanol comprising:
  • a bioreactor (2) in communication with the medium storage tank (1) for fermentation
  • a gas disperser (9) for supplying bubbles into the fermentation broth
  • a membrane separation device (4) in gaseous communication with the bioreactor (2), receiving bubbles from the bioreactor entrained butanol, acetone and ethanol or entrained butanol and membrane separating the butanol, acetone and ethanol or The butanol;
  • a vacuum gauge (6) and a vacuum pump (8) for providing a driving force for the permeation of butanol, acetone and ethanol in gaseous form or in gaseous form;
  • Product storage tank (7) for receiving products is a product storage tank (7) for receiving products.
  • Apparatus according to 8 further comprising a pump (3) for conveying the gas in the membrane separation unit (4) back to the bioreactor (2).
  • the membrane in the membrane separation device (4) is a vaporized permeable membrane, preferably a butoxide and acetone highly selective organic hydrophobic separation membrane or an organic-inorganic composite membrane, more preferably selected from silicone rubber.
  • a vaporized permeable membrane preferably a butoxide and acetone highly selective organic hydrophobic separation membrane or an organic-inorganic composite membrane, more preferably selected from silicone rubber.
  • a method for separating and purifying butanol, acetone and ethanol (or butanol) by a fermentation coupled vaporization permeation technique capable of achieving the above object which first cultures acetone butanol producing bacteria or butanol producing bacteria, and then Fermenting with the above-mentioned production bacteria to obtain butanol, acetone and ethanol (or butanol);
  • the vaporization infiltration method is used to separate and purify butanol, acetone and ethanol (or butanol) from the fermentation liquid, and the specific steps of the vaporization infiltration method are as follows:
  • the butanol, acetone and ethanol (or butanol) in the fermentation broth is subjected to the extraction of butanol, acetone and ethanol (or butanol) into the fermentation system to make butanol, acetone and ethanol (or butanol).
  • Vaporization Wherein the source of the bubble may be a circulation between the fermentation system and the membrane separation device by the produced gas produced by the production bacteria; then the gas entrains the vaporized butanol, acetone and ethanol or the vaporized butanol into the membrane separation device and the membrane.
  • One side is contacted, and a vacuum is applied to the other side of the membrane to vaporize the butanol, acetone, and ethanol (or butanol) through the membrane, condensing or recovering directly into the next stage separation unit.
  • the acetone butanol-producing bacterium is preferably Clostridium acetobutylicum, Clostridium clostridium or other acetone-butanol-producing strain, and genetically engineered bacteria producing only butanol.
  • the acetone butanol producing bacteria or butanol producing bacteria fermentation system comprises a bioreactor, a gas disperser at the bottom of the bioreactor, wherein the gas disperser is a gas that can be integrated with the bioreactor or placed at the bottom of the bioreactor Dispersed parts.
  • the gas used is a self-produced gas such as carbon dioxide and hydrogen produced by the production bacteria in the fermentation process
  • the raw material liquid is a fermentation liquid of acetone butanol producing bacteria containing butanol, acetone and ethanol or a butanol-containing fermentation liquid.
  • the fermentation broth of butanol-producing bacteria, the gas enters the fermentation system to form bubbles to vaporize butanol, acetone and ethanol (or butanol); it can also vaporize butanol, acetone and ethanol (or butanol) with an external gas such as nitrogen. .
  • the vaporization permeable membrane is a high-selective organic hydrophobic membrane or an organic-inorganic composite membrane of butanol and acetone
  • the vaporized permeate raw material liquid is a fermentation liquid containing butanol, acetone and ethanol (or butanol)
  • the membrane is permeabilized.
  • the liquid passing device condenses or the liquid nitrogen directly condenses, the temperature of the device is -30-+15 ° C when condensing, and the vacuum degree is maintained at ⁇ 5 kPa on the side of the membrane permeate.
  • the production, separation and purification efficiency of butanol, acetone and ethanol are effectively improved without increasing equipment investment and saving purification energy consumption, and the liquid organism mainly producing butanol and acetone by biological method is currently used.
  • the production and separation and purification of fuels and bio-based chemicals provide new technical support.
  • the vaporization permeation coupling fermentation technique of the present invention has the advantages of: 1. by bubbling to the bioreactor, entraining butanol, acetone and ethanol (or butanol) without doping other impurities such as proteins, cells, sugar, etc. 2.
  • the vaporized solvent containing butanol, acetone and ethanol (or butanol) has no impurities, so there is no pollution to the membrane, completely solve the problem of membrane fouling; 3.
  • Vaporization osmotic coupling fermentation technology is better than stripping, pervaporation and other technologies It has higher selectivity to butanol, so it can be separated and purified to obtain a higher concentration of concentrated solution of butanol, acetone and ethanol (or butanol); 4.
  • the energy consumption is lower than that of stripping or pervaporation. So far, no reports and related patents have been used to separate and purify butanol and acetone using vaporization infiltration.
  • FIG. 1 is a schematic view showing the structure of a device for producing and separating and purifying butanol, acetone and ethanol (or butanol) for vaporization osmotic coupling fermentation.
  • 1 is a medium storage tank
  • 2 is a bioreactor
  • 3 is a pump
  • 4 is a membrane separation device
  • 5 is a condensation device
  • 6 is a vacuum gauge
  • 7 is a product storage tank
  • 8 is a vacuum pump
  • 9 is a gas disperser.
  • Figure 2 is a schematic illustration of a gas disperser installed in the bottom of a bioreactor of the present invention.
  • the invention relates to a method for producing and separating and purifying butanol, acetone and ethanol (or butanol), which first cultures acetone butanol producing bacteria or butanol producing bacteria, and then ferments the same to obtain butanol and acetone.
  • both sides of the separation membrane are butanol, acetone and ethanol (or butanol) in gaseous form.
  • a seed culture medium is used to culture acetone butanol-producing bacteria.
  • the acetone butanol-producing bacteria are not particularly limited, and examples thereof include production of acetone butanol by Clostridium acetobutylicum, Clostridium beijerinckii, Escherichia coli, and Clostridium tyrobutyricum.
  • the engineered bacteria or the genetically engineered bacteria thereof are preferably Clostridium acetobutylicum.
  • the seed culture medium Before the seed culture medium is used, it is preferably subjected to oxygen removal treatment by introducing nitrogen gas or other inert gas for 10 minutes, and then sterilized at 121 ° C for 30 minutes, and after cooling to room temperature, the acetone butanol production bacteria are added.
  • the acetone butanol producing bacteria are cultured to the most active logarithmic growth phase.
  • the culture time is preferably 12 to 18 hours; the culture temperature is preferably 35 to 39 ° C, more preferably 37 ° C.
  • the seed liquid containing the acetone butanol-producing bacteria obtained in the above step was introduced into the fermentation medium in the bioreactor (fermenter) of Fig. 1 from the seed culture tank, and fermentation was started.
  • the fermentation medium is a substance that provides nutrients (carbon source) for the acetone butanol producing bacteria, and may be glucose as a carbon source in the fermentation medium, or may be corn starch, tapioca starch, molasses, sucrose, tapioca or straw cellulose hydrolyzate ( For example, a straw hydrolyzate or the like is used as a carbon source in the fermentation medium.
  • the fermentation medium is preferably sterilized at 121 ° C for 30 minutes before being introduced into the acetone butanol producing bacteria seed, and subjected to oxygen removal treatment by introducing nitrogen or other inert gas for 2 hours, and after cooling to room temperature, the acetone butanol is produced. bacteria.
  • the amount of the acetone butanol producing bacteria can be appropriately adjusted depending on the amount of the fermentation medium, and is generally 5-10% by volume of the medium.
  • the fermentation temperature is preferably from 35 to 39 ° C, more preferably from 37 ° C.
  • the pH during the fermentation is preferably controlled to be 5.0 or more. When the pH is lower than 5.0, an aqueous sodium hydroxide solution or ammonia water is added to the medium, and when the pH is more than 5.0, no adjustment is required. It is also possible to control the pH in the fermentation process so that the pH in the fermentation broth is freely changed with the fermentation time.
  • the butanol, acetone and ethanol in the fermentation broth are first entrained by bubbles.
  • the bioreactor is used to separate and purify butanol, acetone and ethanol from the fermentation liquid, that is, while fermenting to produce butanol, acetone and ethanol, and separating butanol and butanol from the fermentation liquid (fermentation coupled vaporization infiltration), thereby
  • the butanol, acetone and ethanol in the fermentation broth are continuously removed, and the toxicity of the butanol, acetone and ethanol to the cells is lowered, and the production intensity of the fermentation reaction is increased.
  • the vaporized form of butanol, acetone and ethanol entrained in the bubble can concentrate and purify butanol, acetone and ethanol.
  • the gas pump is started to cause the bubbles to enter the bioreactor, and the gas is circulated in the bioreactor and the membrane separation device to bring the membrane into contact with the gas, and at the same time
  • a cryogenic cooling device is activated on the other side of the membrane to cool the gaseous form of butanol, acetone and ethanol through the membrane, and the vacuum device is activated to provide a driving force for the permeation of butanol, acetone and ethanol in gaseous form.
  • the gas used to extract the butanol and acetone into the fermentation system is preferably any self-produced gas produced by the acetone butanol producing bacteria during the fermentation, usually carbon dioxide and hydrogen, which circulates the gas between the bioreactor and the membrane separation device, usually No external gas is needed, which saves input costs.
  • the butanol concentration in the condensate obtained by the separation of the membrane permeation side can be increased to about 20% to 30% (w/v), and the acetone concentration can be increased to about 10% to 15% (w/v).
  • the conditions for bubbling into the bioreactor in the gasification permeation method are preferably as follows: aeration ratio is 0.5-5 vvm, bubbles are introduced from the bottom of the bioreactor, and bubbles can enter the bioreactor through the gas disperser and exit from the top of the reactor. The bubbles entrained gasified butanol, acetone and ethanol into the membrane separation unit.
  • the separation membrane is fixed in the membrane separation device, one side of the membrane is gasified butanol, acetone and ethanol; the vacuum pump is activated to form a certain degree of vacuum on the other side of the membrane, and substances such as butanol, acetone, ethanol and water are selectively
  • the vapor is passed through the membrane and the permeated vapor is condensed in a product storage tank through a condensing unit to recover a concentrated liquid containing butanol, acetone and ethanol.
  • the concentrated liquid of butanol, acetone and ethanol obtained by the gasification and permeation method can obtain pure products of butanol, acetone and ethanol by distillation, membrane separation, molecular sieve purification and the like.
  • the temperature of the membrane separation device is 0-80 °C. Condensation of membrane permeate is usually selected from condensed water or liquid nitrogen. Condensation can also be used for condensation. If condensation is selected or condensation is condensed, the condensation temperature is -30-+. At 15 ° C, the degree of vacuum on the permeate side of the membrane is preferably from 0 to 10 kPa.
  • the gasification osmosis coupled fermentation of the present invention achieves the dual purpose of continuous removal and effective recovery and concentration of the toxicity inhibiting products butanol, acetone and ethanol (or butanol) during fermentation, and both sides of the membrane
  • the method improves the production efficiency of butanol, acetone and ethanol (or butanol), reduces the recovery cost of butanol, acetone and ethanol (or butanol), and improves the production of butanol, acetone and ethanol by fermentation ( Or butanol) economic benefits, suitable for popularization and application.
  • the present invention will be specifically described by taking the acetone butanol producing bacteria as an example in conjunction with the examples.
  • the present invention is not limited by the following examples, and the present invention can be appropriately modified within the scope of the present invention.
  • the experimental methods used are all conventional methods, and materials, reagents and the like used can be purchased from a biological or chemical company.
  • Acetobutanol-producing bacteria Clostridium acetobutylicum, purchased from the ATCC strain library (ATCC number: 55025-E604).
  • Pretreatment method of corn stalk smash corn stalk, pass 0.4mm sieve, take 200g smashed corn stalk and add it to 2L 2% (w/v) NaOH solution, react at 120 °C for 30min, cool to room temperature. Filter, wash the filter residue to neutral, and dry at 50 °C.
  • the corn stalk processed as described above was separately added to the cellulase liquid at a weight ratio of 1:10, the concentration of the cellulase liquid was 0.03 mol/L, and the enzyme buffer used was citric acid-sodium citrate.
  • the pH of the enzyme buffer is 4.8
  • the enzyme activity of the cellulase is 20 FPU/g
  • the rotation speed is 150 r/min
  • the enzymolysis is carried out at 50 ° C for 72 h
  • the centrifugation is performed at 8000 r/min for 5 min after the end of enzymatic hydrolysis to obtain a supernatant.
  • the supernatant was adjusted to pH 6.2 with concentrated aqueous ammonia, and 0.115% (w/v) of yeast dipping powder and mineral mixture was added.
  • the mineral mixture contained 0.2 g/L of 7-hydrate magnesium sulfate, 7-hydrate sulfuric acid. Iron 0.01g / L, manganese sulfate 0.01g / L, and sodium chloride 0.01g / L, sterilized by introducing nitrogen for 5min.
  • Preparation of corn starch saccharification solution the corn flour is mixed with warm water of about 60-65 °C in a ratio of 1:2.5, and ⁇ -amylase is added in a ratio of 0.6 ml per kg of corn flour, and the temperature is raised to 85-90 ° C. Liquefaction for 1-2 hours, then cooling to 60-65 ° C, adding glucoamylase in a ratio of 1.2 ml per kg of corn flour, saccharification for 10-15 hours, and filtering to obtain a saccharification solution.
  • the saccharification solution can be diluted with 1:3 plus water for fermentation.
  • Seed medium 30 g of glucose, 2 g of yeast powder, 4 g of tryptone, 0.5 g of potassium dihydrogen phosphate, 0.5 g of dipotassium hydrogen phosphate, 2.2 g of ammonium acetate, and a mineral mixture per liter of the medium.
  • the composition of the mineral mixture is: 0.1 g of magnesium sulfate 7 hydrate, 0.015 g of ferrous sulfate 7 hydrate, 0.015 g of calcium chloride 2 hydrate, 0.01 g of manganese sulfate monohydrate, and 0.02 g of cobalt chloride per liter of the medium. And 0.002 g of zinc sulfate.
  • a medium in which glucose is a carbon source 80 g of glucose per liter of medium, 1 g of yeast powder, 0.5 g of potassium dihydrogen phosphate, 0.5 g of dipotassium hydrogen phosphate, 2.2 g of ammonium acetate, a mineral mixture, and a vitamin.
  • the composition of the mineral mixture is: 0.2 g of magnesium sulfate 7 hydrate, 0.01 g of ferrous sulfate 7 hydrate, 0.01 g of manganese sulfate monohydrate and 0.01 g of sodium chloride per liter of the medium;
  • the composition of the vitamin is: per liter
  • the medium contained 0.001 g of p-aminobenzoic acid, 0.001 g of vitamin B1, and 0.00001 g of biotin.
  • composition of the fermentation medium containing corn stalk hydrolysate as carbon source 48.4 g of glucose per glucose, 15.6 g of xylose, 4.8 g of cellobiose, 2.4 g of arabinose, 1 g of yeast powder, potassium dihydrogen phosphate 0.5 g, 0.5 g of dipotassium hydrogen phosphate, 2.2 g of ammonium acetate, 0.2 g of magnesium sulfate 7-hydrate, 0.01 g of ferrous sulfate 7-hydrate, 0.01 g of manganese sulfate monohydrate, 0.01 g of sodium chloride, and 0.001 g of p-aminobenzoic acid. Vitamin B1 0.001g and biotin 0.00001g.
  • Fermentation medium consisting of corn starch as carbon source: 200g of reducing sugar per liter of medium, 0.5g of potassium dihydrogen phosphate, 0.5g of dipotassium hydrogen phosphate, 2.2g of ammonium acetate, 0.2g of magnesium sulfate 7-hydrate, 7 Hydrated ferrous sulfate 0.01 g, manganese sulfate monohydrate 0.01 g, sodium chloride 0.01 g, p-aminobenzoic acid 0.001 g, vitamin B1 0.001 g, and biotin 0.00001 g.
  • the reducing sugar concentration of the corn starch medium at the initial stage of fermentation can be adjusted at 60-100 g/L by dilution with water.
  • the concentration of reducing sugar in the corn starch fermentation medium can be 150-200g/L.
  • the seed medium was deoxygenated with nitrogen for 10 minutes before use, and then sterilized at 121 ° C for 30 minutes, cooled to room temperature, and then introduced into the production bacteria.
  • the culture bacteria were cultured in the seed culture tank 1 at 37 ° C for 15 hours, and then prepared for access to the fermentation medium.
  • the fermentation medium was sterilized at 121 ° C for 30 minutes before use, and then deoxygenated by nitrogen for 2 h.
  • the seed liquid containing the production bacteria (10% of the fermentation medium volume) was pumped into the biological reaction by a pump.
  • the fermentation was started at 37 ° C in the apparatus.
  • the fermentation medium is initially not adjusted in pH value, and when the pH of the fermentation liquid is lower than 5.0, an aqueous sodium hydroxide solution or ammonia water is automatically added to adjust the pH to 5.0 or more.
  • the bioreactor can be an agitated bioreactor, an airlift bioreactor, or a stationary bioreactor.
  • PDMS Polydimethylsiloxane
  • ZSM-5 Zeolite Nanomaterials
  • ZSM-5 was first dried at 80 ° C for 24 hours.
  • the gelatin base liquid and the curing agent in the polydimethylsiloxane (PDMS) are mixed in a ratio of 10:1.
  • centrifugation was carried out directly at 8000 rpm for 5 minutes for subsequent operations.
  • the specified weight ratio (20%-80%) of ZSM-5 was mixed with the PDMS membrane solution mixed at a ratio of 10:1, and centrifuged at 8000 rpm for 5 minutes.
  • the subsequent operation is to first treat the film forming solution with ultrasonic for 15 minutes, remove the air bubbles in the film forming liquid, and then uniformly apply the film forming liquid to the glass plate with a doctor blade, and place the glass plate with the film forming liquid in an oven at 100 ° C. Film formation in 3 hours. Finally, the glass plate was taken out from the oven, and the prepared pervaporation gasification film was peeled off and fixed in a membrane separation device for gasification osmotic coupling separation and purification operation.
  • the fermentation and fermentation of acetone butanol producing bacteria were carried out by using the fermentation medium in which glucose was used as a carbon source as described above. When the production bacteria are connected to the bioreactor, fermentation is started until the end of the fermentation.
  • the results are shown in Table 1.
  • the endpoint concentrations of butanol, acetone and ethanol in the fermentation broth were 14 g/L (1.4%), 7 g/L (0.7%) and 2 g/L (0.2%).
  • the cultivation and fermentation of the acetone butanol producing bacteria were carried out as described above.
  • fermentation begins.
  • the conventional method performs gas stripping coupling fermentation.
  • the gas stripping separation device is started.
  • Conventional stripping coupled fermentation processes vaporize butanol, acetone and ethanol by bubbling to the bioreactor.
  • the gas and vaporized butanol, acetone and ethanol are passed through a condenser to condense and recover the butanol, acetone and ethanol in a liquid form, and the condensation tube is not purified and concentrated.
  • Table 1 The results are shown in Table 1.
  • the concentration of butanol recovered in the condensate obtained by stripping is 9.0%-11% (w/v), the acetone concentration is 3.5%-5.0% (w/v), and the ethanol concentration is 0.5%-1.5% (w/ v).
  • the change in the concentration of butanol, acetone and ethanol in the condensate recovered by stripping is mainly caused by the change in the concentration of butanol, acetone and ethanol in the fermentation broth.
  • the cultivation and fermentation of the acetone butanol producing bacteria were carried out as described above.
  • fermentation begins.
  • the traditional method is carried out by permeation gasification coupled fermentation.
  • the pervaporation membrane separation device is started, and the separation membrane is a pure PDMS polymerization membrane.
  • the pervaporation coupled fermentation process circulates the liquid form of the fermentation broth between the bioreactor and the pervaporation membrane separation unit.
  • the fermentation broth passes through one side of the separation membrane, the fermentation liquid in liquid form passes through the separation membrane in a dissolved diffusion manner, and the fermentation liquid is concentrated and purified.
  • Table 1 The traditional results are shown in Table 1.
  • the separation membrane is separated and purified by PDMS/50% ZSM-5 mixed membrane.
  • the concentration of butanol, acetone and ethanol in the condensate obtained by pervaporation separation is 14.0%-18.0% (w/v), 6.0%-9.0. %(w/v) and 1.0%-2.0%(w/v).
  • the change in the concentration of butanol, acetone and ethanol in the condensate recovered by the pervaporation method is mainly caused by the change in the concentration of butanol, acetone and ethanol in the fermentation broth.
  • Example 1 Gasification osmotic coupling fermentation using glucose as a carbon source to produce butanol, acetone and ethanol
  • the cultivation and fermentation of the acetone butanol producing bacteria were carried out as described above.
  • fermentation begins.
  • the butanol concentration in the fermentation broth reaches 5 g/L
  • the pump between the bioreactor and the membrane separation device is turned on, the bubble is bubbled into the bioreactor, and the gas and gas entrained in the vaporized state of butanol and acetone are pumped.
  • ethanol circulate in a closed space formed by the bioreactor and the membrane separation device.
  • the gasified state of butanol, acetone and ethanol passes through the side of the separation membrane, it dissolves and diffuses to the other side of the membrane, and is condensed and concentrated to be recovered. .
  • the glucose concentration in the fermentation medium was lowered to 10 g/L or less, 400 g/L of concentrated glucose was added to the fermentation medium to continue the fermentation. Since butanol, acetone and ethanol are continuously recovered from the bioreactor by gasification and permeation, there is sufficient glucose carbon source in the fermentation medium, and the fermentation can be stably operated for more than 150 hours.
  • the concentration of butanol, acetone and ethanol in the condensate obtained by gasification permeation method is 20.0%-24.0% (w/v), 8.0%-12.0% (w/v) and 0.8%- 1.2% (w/v).
  • the separation and purification were carried out by using PDMS/50% ZSM-5 mixed membrane.
  • the concentration of butanol, acetone and ethanol in the condensate obtained by gasification and permeation method was 28.0%-32.0% (w/v), 11.0%-15.0% (w). /v) and 1.0%-2.0% (w/v). It can be seen that compared with the stripping coupling fermentation of Comparative Example 2, the concentration of butanol and acetone in the condensate obtained by the gasification permeation method is more than doubled, and the concentration of ethanol is increased by about one time. Compared with the pervaporation of Comparative Example 3, the concentration of butanol and acetone in the condensate obtained by the gasification permeation method was increased by about 2 times, and the concentration of ethanol was hardly changed.
  • hydrophobic particles ZSM-5 to the PDMS membrane can increase the separation and purification concentration of butanol and acetone, and improve the selectivity of the membrane.
  • addition of other nano-hydrophobic particles to the membrane can also achieve the effect of increasing the separation efficiency of butanol and acetone.
  • the cultivation and fermentation of the acetone butanol producing bacteria were carried out as described above.
  • fermentation begins.
  • the butanol concentration in the fermentation broth reaches 5 g/L
  • the pump between the bioreactor and the membrane separation device is turned on, the bubble is bubbled into the bioreactor, and the gas and gas entrained in the vaporized state of butanol and acetone are pumped.
  • ethanol circulate in a closed space formed by the bioreactor and the membrane separation device.
  • the gasified state of butanol, acetone and ethanol passes through the side of the separation membrane, it dissolves and diffuses to the other side of the membrane, and is condensed and concentrated to be recovered. .
  • the corn stover hydrolyzate is added to the fermentation medium to continue the fermentation. Since butanol, acetone and ethanol are continuously recovered from the bioreactor by gasification and permeation, there is sufficient carbon source in the fermentation medium to supplement, and the fermentation can continue to operate stably.
  • concentration of butanol, acetone and ethanol in the condensate obtained by gasification permeation method is 14.0%-18.0% (w/v), 6.0%-9.0% (w/v) and 0.6%- 1.0% (w/v).
  • the separation and purification were carried out by using PDMS/50% ZSM-5 mixed membrane.
  • the concentration of butanol, acetone and ethanol in the condensate obtained by gasification and permeation method was 18.0%-22.0% (w/v), 7.0%-11.0% (w). /v) and 1.0%-1.5% (w/v). It can be seen that compared with the stripping-coupled fermentation of Comparative Example 2 and the pervaporation-coupled fermentation of Comparative Example 3, the concentrations of butanol and acetone in the condensate obtained by the gasification and permeation method are much higher, indicating that the gasification and permeation coupling fermentation can be Separation and purification yield higher concentrations of butanol and acetone, which have higher selectivity and better separation of butanol and acetone products.
  • Example 3 Production of butanol, acetone and ethanol by gasification osmotic coupling fermentation using corn starch saccharification solution as carbon source
  • the cultivation and fermentation of the acetone butanol producing bacteria were carried out as described above.
  • fermentation begins.
  • the butanol concentration in the fermentation broth reaches 5 g/L
  • the bioreactor and the membrane separation device are opened.
  • a pump that bubbling into the bioreactor the pump circulates the vaporized state of butanol, acetone, and ethanol entrained by the gas and gas in a closed space formed by the bioreactor and the membrane separation device, when the butanol in the vaporized state,
  • acetone and ethanol pass through one side of the separation membrane, they dissolve and diffuse to the other side of the membrane, and are concentrated by condensation.
  • the corn starch saccharification solution is added to the fermentation medium to continue the fermentation. Since butanol, acetone and ethanol are continuously recovered from the bioreactor by gasification and permeation, there is sufficient carbon source in the fermentation medium, and the fermentation can continue to operate stably.
  • concentration of butanol, acetone and ethanol in the condensate obtained by gasification permeation method is 17.0%-22.0%% (w/v), 7.0%-11.0% (w/v) and 0.7%. 1.1% (w/v). Separation and purification were carried out using PDMS/50% ZSM-5 mixed membrane.
  • the concentration of butanol, acetone and ethanol in the condensate obtained by gasification permeation method was 22.0%-30.0% (w/v), 9.0%-14.0% (w /v) and 1.0%-1.7% (w/v). It can be seen that compared with the stripping-coupled fermentation of Comparative Example 2 and the pervaporation-coupled fermentation of Comparative Example 3, the concentrations of butanol and acetone in the condensate obtained by the gasification and permeation method are much higher, indicating that the gasification and permeation coupling fermentation can be Separation and purification yield higher concentrations of butanol and acetone, which have higher selectivity and better separation of butanol and acetone products.
  • the concentration of butanol and acetone in the fermentation broth obtained by conventional uncoupled separation of acetone butanol is 1.4%, 0.7% and 0.2%, respectively.
  • the concentration of butanol and acetone in the condensate obtained by the conventional one-step stripping method is about 9.0% to 11.0%, 3.5% to 5.0%, and 0.5% to 1.5%, respectively.
  • the concentration of butanol, acetone and ethanol in the condensate obtained by pervaporation separation is 14.0%-18.0% (w/v), 6.0%-9.0% (w/v) and 1.0%-2.0% (w/v). .
  • the product finally collected by the gasification osmotic coupling fermentation technology contains the concentrations of butanol, acetone and ethanol of 28.0%-32.0%, 11.0%-15.0% and 1.0%-2.0%, respectively. It can be seen that the gasification osmotic coupling fermentation technique can obtain the product concentration of the highest purification separation. Since the final product contains very high concentrations of butanol and acetone, it is easy to obtain pure butanol and acetone by a simple dehydration treatment such as distillation, distillation or membrane separation.
  • the gasification and permeation process of the present invention is essential for increasing the concentration of butanol purification and reducing the energy consumption of the entire fermentation separation process.
  • the gasification permeation method requires about 30% of the energy consumption of conventional rectification separation. Moreover, it is ensured that the subsequent purification of butanol and acetone can be carried out in a solution rich in high concentrations of butanol and acetone with low energy consumption and high efficiency. More importantly, the side of the pervaporation membrane requires the fermentation broth to contact the membrane as compared to conventional vaporization osmotic coupling fermentation. Microbial cells, carbohydrates, proteinaceous substances and other macromolecular substances in the fermentation broth can contaminate the membrane during long-term operation of the membrane, and the cost of cleaning and replacing the membrane is very high.
  • the vaporization and permeation technology used in the invention is coupled to the fermentation, and both sides of the membrane are organic solvents in a vaporized state, which does not pollute the membrane and ensures the service life of the membrane. Therefore, the invention can improve the production and recovery efficiency of butanol and acetone and reduce the energy consumption of separation and purification, and provides a new technology for producing butanol and acetone by biological method, and has great industrial application value.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Health & Medical Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Microbiology (AREA)
  • Water Supply & Treatment (AREA)
  • Biotechnology (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Inorganic Chemistry (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Geology (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

提供了一种发酵耦合分离纯化丁醇、丙酮和乙醇或纯化丁醇的方法，包括以下步骤：1)利用丙酮丁醇生产菌发酵得到丁醇、丙酮和乙醇或利用丁醇生产菌得到丁醇；2)采用汽化渗透法从发酵液在线分离纯化丁醇、丙酮和乙醇或纯化丁醇。本发明的方法提高了丁醇、丙酮和乙醇或丁醇的生产和分离提纯效率。

Description

一种发酵耦合分离纯化生产丁醇的方法及装置 技术领域
本发明涉及一种发酵耦合分离纯化生产丁醇、丙酮和乙醇(或丁醇)的方法，属于生物技术领域。
背景技术
丁醇和丙酮作为一种潜在的液体能源和应用于医药和食品行业的重要化学品，可以通过微生物发酵法获得，详见文献：Dürre,P.Biobutanol:an attractive biofuel.Biotechnol.J.2：1525–1534，2007。但是用丙酮丁醇梭菌或拜式梭菌发酵生产丁醇时，发酵液中终点的丁醇浓度通常不超过2.0％(w/v)。并且，丁醇的沸点为117.7℃，高于水的沸点100℃。因此，如果利用传统的精馏或蒸馏分离法，其分离成本极高，经济上是不可行，很难实现工业化生产(Matsumura,M.,Kataoka,H.,Sueki,M.,Araki,K.Energy saving effect of pervaporation using oleyl alcohol liquid membrane in butanol purification.Bioprocess Eng.3：93-100，1988)。
可替代性分离技术如液-液萃取、气提、吸附和渗透汽化等，可以通过在发酵过程中不断移除并回收对细胞产生抑制的产物丁醇，可以提高发酵效率，是提高生物法生产丁醇的有效技术(Xue C,Zhao JB,Chen LJ,Bai FW,Yang ST,Sun JX.Integrated butanol recovery for an advanced biofuel:current state and prospects.Appl Microbiol Biotechnol,2014,98:3463–3474.；Qureshi,N.,Meagher,M.M.,Huang,J.C.,Hutkins,R.W.Acetone butanol ethanol(ABE)recovery by pervaporation using silicalite-silicone composite membrane from fed-batch reactor of Clostridium acetobutylicum.J.Membr.Sci.187：93-102,2001)。但是目前利用这些分离技术进行分离，主要的问题是分离效率低，分离产物浓度低，仍然需要进一步对技术进行优化和对产物进行脱水处理。
发明内容
基于现有技术中存在的上述问题，本发明利用汽化渗透法耦合发酵生产丁醇、丙酮和乙醇(或丁醇)，在发酵的过程中在线分离纯化丁醇、丙酮和乙醇(或丁醇)。
本发明具体涉及以下各项：
1.一种发酵耦合分离纯化丁醇、丙酮和乙醇或纯化丁醇的方法，所述方法包括以下步骤：
1)利用丙酮丁醇生产菌或丁醇生产菌发酵得到丁醇、丙酮和乙醇或得到丁醇；
2)采用汽化渗透法从发酵液在线分离纯化丁醇、丙酮和乙醇；
其中所述汽化渗透法包括以下步骤：
a.向发酵液中通入气泡，使丁醇、丙酮和乙醇或使丁醇汽化；
b.使气泡中的气体夹带汽化的丁醇、丙酮和乙醇或汽化的丁醇进入膜分离装置内并透过膜；
c.回收所述丁醇、丙酮和乙醇或回收所述丁醇，或进入下一级分离装置。
2.根据1所述的方法，其中所述气泡的来源为丙酮丁醇生产菌或丁醇生产菌的自产气体或外源气体，所述自产气体优选为二氧化碳和/或氢气，所述外源气体优选为氮气。
3.根据1所述的方法，其中所述丙酮丁醇生产菌优选选自丙酮丁醇梭菌(Clostridium acetobutylicum)、拜式梭菌(Clostridium beijerinckii)、大肠杆菌或酪丁酸梭菌(Clostridium tyrobutyricum)以及这些菌株的基因工程改造菌，优选为丙酮丁醇梭菌，所述丁醇生产菌为产丁醇的基因工程菌，优选为产丁醇大肠杆菌或产丁醇梭菌。
4.根据1所述的方法，其中所述膜为汽化渗透膜，优选为丁醇和丙酮高选择性有机疏水分离膜或有机无机复合膜，更优选选自硅橡胶、聚三甲基硅丙炔、聚丙烯、聚丁二烯、聚偏氟乙烯、聚四氟乙烯或其衍生物、丁腈橡胶、或分子筛材料中的至少一种，最优选为含PDMS的复合膜，所述膜的类型优选为管式、卷式、平板或中空纤维膜形式。
5.根据1所述的方法，其中所述回收通过冷凝进行，所述冷凝优选用装置冷凝或液氮或冷凝水冷凝，其中装置冷凝的温度为-30-+15℃。
6.根据1所述的方法，其中所述气泡从生物反应器底部通入，通气比为0.5-5vvm。
7.根据1所述的方法，所述膜分离装置的膜透过液一侧保持真空度为0-10kpa，优选<5kpa。
8.一种用于发酵耦合分离纯化丁醇、丙酮和乙醇或纯化丁醇的装置，其包括:
培养基储罐(1)，用于向生物反应器提供培养基；
生物反应器(2)，与所述培养基储罐(1)连通，用于发酵；
气体分散器(9)，用于向发酵液中提供气泡；
膜分离装置(4)，与所述生物反应器(2)气体连通，接收来自生物反应器的夹带丁醇、丙酮和乙醇或夹带丁醇的气泡并膜分离所述丁醇、丙酮和乙醇或所述丁醇；
冷凝装置(5)，用于回收所述丁醇、丙酮和乙醇或所述丁醇；
真空表(6)和真空泵(8)，用于为气态形式的丁醇、丙酮和乙醇或气态形式的丁醇透过提供驱动力；
产品储罐(7)，用于接收产品。
9.根据8所述的装置，其还包括泵(3)，用于将膜分离装置(4)中的气体输送回所述生物反应器(2)。
10.根据8所述的装置，其中所述膜分离装置(4)中的膜为汽化渗透膜，优选为丁醇和丙酮高选择性有机疏水分离膜或有机无机复合膜，更优选选自硅橡胶、聚三甲基硅丙炔、聚丙烯、聚丁二烯、聚偏氟乙烯、聚四氟乙烯或其衍生物、丁腈橡胶、或分子筛材料中的至少一种，最优选为含PDMS的复合膜，所述膜的类型优选为管式、卷式、平板或中空纤维膜形式。
具体而言，本发明的目的在于提供一种高效生产并分离纯化丁醇、丙酮和乙醇或高效生产并分离纯化丁醇的方法及相应装置。特别是一种能够实现上述目的的本发明一种发酵耦合汽化渗透技术分离纯化丁醇、丙酮和乙醇(或丁醇)的方法，该方法先培养丙酮丁醇生产菌或丁醇生产菌，再用上述生产菌发酵得到丁醇、丙酮和乙醇(或丁醇)； 采用汽化渗透法从发酵液在线分离纯化丁醇、丙酮和乙醇(或丁醇)，其特征是汽化渗透法具体步骤如下：
发酵液中的丁醇、丙酮和乙醇(或丁醇)汽化方法采用向发酵系统中通入气泡提取丁醇、丙酮和乙醇(或丁醇)，使丁醇、丙酮和乙醇(或丁醇)汽化。其中气泡来源可以采用所述生产菌产生的自产气体在发酵系统和膜分离装置之间的循环；然后气体夹带汽化的丁醇、丙酮和乙醇或汽化的丁醇进入膜分离装置内与膜的一侧接触，在膜的另一侧抽真空，使汽化的丁醇、丙酮和乙醇(或丁醇)透过膜，冷凝回收或直接进入下一级分离装置。
在本发明的方法中，丙酮丁醇生产菌优选为丙酮丁醇梭菌、拜式梭菌或者其他丙酮丁醇生产菌株以及只产丁醇的基因工程菌。
优选丙酮丁醇生产菌或丁醇生产菌发酵系统包括生物反应器，生物反应器底部的气体分散器，其中气体分散器是可以与生物反应器为一体或放置在生物反应器底部的可使气体分散的部件。
在汽化渗透法中优选：所用的气体为发酵过程中生产菌产生的自产气体如二氧化碳和氢气，原料液为含有丁醇、丙酮和乙醇的丙酮丁醇生产菌的发酵液或含有丁醇的丁醇生产菌的发酵液，气体进入发酵系统中形成气泡使丁醇、丙酮和乙醇(或丁醇)汽化；也可以利用外源气体如氮气使丁醇、丙酮和乙醇(或丁醇)汽化。
在汽化渗透法中优选：汽化渗透膜为丁醇和丙酮高选择性有机疏水膜或有机无机复合膜，汽化渗透的原料液为含有丁醇、丙酮和乙醇(或丁醇)的发酵液，膜透过液用装置冷凝或液氮直接冷凝，装置冷凝时温度为-30-+15℃，在膜透过液一侧保持真空度在<5kpa。
通过本发明的方法，在不增加设备投资和节省提纯能耗的前提下，有效提高了丁醇、丙酮和乙醇的生产和分离提纯效率，为目前以生物法生产丁醇和丙酮为主的液体生物燃料和生物基化学品的生产和分离提纯提供了新的技术支持。
本发明中的汽化渗透法耦合发酵技术的优势在于，1.通过向生物反应器鼓泡，夹带出丁醇、丙酮和乙醇(或丁醇)，不掺杂其他杂质如蛋白质，细胞，糖等原料；2.汽化的含丁醇、丙酮和乙醇(或丁醇)的溶剂没有杂质，因此对膜没有污染，彻底解决膜污染问题；3.汽化渗透耦合发酵技术比气提，渗透汽化等技术对丁醇有更高的选择性，因此可以分离纯化得到更高浓度的丁醇、丙酮和乙醇(或丁醇)的浓缩液；4.能耗低于气提或渗透汽化等技术。到目前为止，未见使用汽化渗透法对丁醇和丙酮进行分离纯化的报道和相关专利。
附图说明
图1为本发明中用于汽化渗透耦合发酵的生产和分离纯化丁醇、丙酮和乙醇(或丁醇)的装置结构示意图。其中1为培养基储罐，2为生物反应器，3为泵，4为膜分离装置，5为冷凝装置，6为真空表，7为产品储罐，8为真空泵，9为气体分散器。
图2为本发明中安装在生物反应器底部的气体分散器示意图。
具体实施方式
本发明是一种生产和分离纯化丁醇、丙酮和乙醇(或丁醇)的方法，该方法先培养丙酮丁醇生产菌或丁醇生产菌，再用所述生产菌发酵得到丁醇、丙酮和乙醇(或丁 醇)；采用汽化渗透法从发酵液在线分离纯化丁醇、丙酮和乙醇(或丁醇)；通过向生物反应器(发酵罐)底部鼓气泡使丁醇、丙酮和乙醇(或丁醇)汽化，随气泡离开生物反应器，所述气泡为管路中的发酵过程中所述生产菌产生的自产气体或外源气体如氮气，气体夹带汽化的丁醇、丙酮和乙醇(或丁醇)进入膜分离装置，与膜的一侧接触，通过在膜的另一侧抽真空，使丁醇、丙酮和乙醇(或丁醇)通过膜，透过膜的丁醇、丙酮和乙醇(或丁醇)，冷凝回收或直接进入下一级分离装置。因此，分离膜的两侧均为气态形式的丁醇、丙酮和乙醇(或丁醇)。
下面，以丙酮丁醇生产菌为例结合附图1对发明进行详细说明。
<培养丙酮丁醇生产菌>
首先，在种子培养罐中，使用种子培养基来培养丙酮丁醇生产菌。
对所述丙酮丁醇生产菌没有特别限制，可列举丙酮丁醇梭菌(Clostridium acetobutylicum)、拜式梭菌(Clostridium beijerinckii)、大肠杆菌和酪丁酸梭菌(Clostridium tyrobutyricum)等生产丙酮丁醇的工程菌或其基因工程改造菌，优选丙酮丁醇梭菌。
所述种子培养基在使用之前，优选先通入氮气或其他惰性气体10分钟进行除氧处理后，再在121℃灭菌30分钟，冷却到室温后，接入丙酮丁醇生产菌。
优选将丙酮丁醇生产菌培养到生长最活跃的对数生长期。为了将丙酮丁醇生产菌培养到对数生长期，培养时间优选为12-18h；培养温度优选为35-39℃，更优选为37℃。
<丙酮丁醇生产菌发酵得到丁醇、丙酮和乙醇>
然后，将上述步骤中得到的含有丙酮丁醇生产菌的种子液从种子培养罐中接入到图1的生物反应器(发酵罐)中的发酵培养基后，开始发酵。
发酵培养基是为丙酮丁醇生产菌提供营养(碳源)的物质，可以葡萄糖作为发酵培养基中的碳源，也可以玉米淀粉、木薯淀粉、糖蜜、蔗糖、木薯或者秸秆纤维素水解液(例如秸秆水解液)等作为发酵培养基中的碳源。
所述发酵培养基在接入丙酮丁醇生产菌种子之前，优选在121℃灭菌30分钟后，通入氮气或其他惰性气体2h进行除氧处理，冷却到室温后，接入丙酮丁醇生产菌。
丙酮丁醇生产菌的接入量可依据发酵培养基的量适当调整，一般为培养基的5-10％(体积百分比)。
发酵温度优选为35-39℃，更优选为37℃。发酵过程中的pH优选控制在5.0以上，当pH低于5.0时，向培养基中加入氢氧化钠水溶液或氨水，当pH大于5.0时，不需要调整。也可以发酵过程中不控制pH，使发酵液中的pH随发酵时间自由变化。
<利用汽化渗透法耦合发酵从发酵液中在线分离纯化丁醇、丙酮和乙醇>
本发明中，对于汽化渗透法，第一，利用丁醇、丙酮和乙醇的挥发性和气体对丁醇、丙酮和乙醇的吸附原理，先通过气泡将发酵液中的丁醇、丙酮和乙醇夹带出生物反应器，从发酵液中在线分离纯化丁醇、丙酮和乙醇，即一边进行发酵产生丁醇、丙酮和乙醇，一边从发酵液中分离纯化丁醇和丁醇(发酵耦合汽化渗透)，从而使发酵液中的丁醇、丙酮和乙醇不断被移除，降低丁醇、丙酮和乙醇对细胞的毒害，使发酵反应的生产强度提高。第二，气泡夹带出来的汽化形式的丁醇、丙酮和乙醇可以使丁醇、丙酮和乙醇浓缩纯化；第三，利用丁醇、丙酮，乙醇和水在汽化渗透膜中溶解和 扩散能力的差别，汽化的丁醇、丙酮和乙醇较多的溶解在膜上，并扩散通过膜，在膜的另一侧被抽出，获得高浓度的丁醇、丙酮和乙醇；第四，汽化渗透膜的两侧均为气态形式的丁醇、丙酮和乙醇，因此对膜没有任何污染。
对于汽化渗透耦合发酵操作过程，当生物反应器中的丁醇达到一定浓度后，启动气泵使气泡进入生物反应器，使气体在生物反应器和膜分离装置中循环，使膜与气体接触，同时在膜的另一侧启动低温冷却装置冷却透过膜的气态形式的丁醇、丙酮和乙醇，启动真空装置为气态形式的丁醇、丙酮和乙醇透过提供驱动力。
进入发酵系统中用于提取丁醇和丙酮的气体优选为发酵过程中丙酮丁醇生产菌产生的任何自产气体，通常是二氧化碳和氢气，使气体在生物反应器和膜分离装置之间循环，通常不需要外源气体，这样就可节省投入成本。
这样，膜透过侧的分离得到的冷凝液中的丁醇浓度可提高到20％-30％(w/v)左右，丙酮浓度提高到10％-15％(w/v)左右。
气化渗透法中向生物反应器中鼓泡的条件优选如下：通气比为0.5-5vvm，气泡从生物反应器底部通入，气泡可通过气体分散器进入生物反应器，从反应器顶部出来，气泡夹带气化的丁醇、丙酮和乙醇进入膜分离装置。
膜分离装置中固定分离膜，膜的一侧为气化的丁醇、丙酮和乙醇；启动真空泵使膜的另一侧形成一定的真空度，丁醇、丙酮，乙醇和水等物质选择性地以蒸汽形式透过膜，透过的蒸汽在产品储罐中通过冷凝装置冷凝回收含有丁醇、丙酮和乙醇的浓缩液体。
气化渗透法得到的丁醇、丙酮和乙醇的浓缩液体，可通过精馏，膜分离，分子筛纯化等方法获得丁醇、丙酮和乙醇的纯品。
膜分离装置的温度条件为0-80℃膜透过液的冷凝通常选取冷凝水或液氮，也可以选取冷凝装置进行冷凝，如果选取冷凝装置冷凝或冷凝水冷凝，冷凝温度在-30-+15℃，膜透过液一侧的真空度优选0-10kpa。
如上所述，本发明的气化渗透法耦合发酵实现了发酵过程中毒性抑制产物丁醇、丙酮和乙醇(或丁醇)的不断移除和有效回收与浓缩的双重目的，并且膜两侧均为洁净的气化形式的丁醇、丙酮和乙醇(或丁醇)，对膜没有任何污染。因此，该方法提高了丁醇、丙酮和乙醇(或丁醇)的生产效率，降低了丁醇、丙酮和乙醇(或丁醇)的回收成本，提高了发酵法生产丁醇、丙酮和乙醇(或丁醇)的经济收益，适合于推广应用。
实施例
下面以丙酮丁醇生产菌为例结合实施例对本发明作具体说明。但本发明不受下述实施例的限制，在符合本发明前后宗旨的范围内，可对本发明作适当变更。另外，下述实施例中，如无特殊说明，所使用的实验方法均为常规方法，所用材料、试剂等均可从生物或化学公司购买。
丙酮丁醇生产菌：丙酮丁醇梭菌(Clostridium acetobutylicum)，购买于美国ATCC菌种库(ATCC number：55025-E604)。
玉米秸秆的预处理方法：将玉米秸秆粉碎、过0.4mm筛子，取200g粉碎过筛后的玉米秸秆加入到2L的2％(w/v)NaOH溶液中，120℃反应30min，冷却至室温， 过滤，将滤渣洗至中性，50℃烘干。将上述处理的玉米秸秆分两次按重量比1：10分别添加到纤维素酶液中，所述纤维素酶液的浓度为0.03mol/L，使用的酶缓冲液为柠檬酸-柠檬酸钠缓冲液，所述酶缓冲液的pH为4.8，所述纤维素酶的酶活力为20FPU/g，转速150r/min，50℃酶解72h，酶解结束后8000r/min离心5min，得到上清液，将上清液用浓氨水调节pH至6.2，加入0.115％(w/v)的酵母浸粉和矿物质混合物，所述矿物质混合物含7水合硫酸镁0.2g/L、7水合硫酸亚铁0.01g/L、1水合硫酸锰0.01g/L和氯化钠0.01g/L，通入氮气5min后灭菌。
玉米淀粉糖化液的制备：将玉米粉与60-65℃左右的温水按1:2.5的比例配制成粉浆，按每公斤玉米粉0.6ml的比例加入α-淀粉酶，升温至85-90℃，液化1-2小时，然后降温至60-65℃，按每公斤玉米粉1.2ml的比例加入糖化酶，糖化10-15小时，过滤得到糖化液。糖化液可按照1:3加水进行稀释，用于发酵。
种子培养基：每升培养基中含葡萄糖30g、酵母粉2g、胰蛋白胨4g、磷酸二氢钾0.5g、磷酸氢二钾0.5g、乙酸铵2.2g和矿物质混合物。其中，矿物质混合物的组成为：每升培养基中含7水合硫酸镁0.1g、7水合硫酸亚铁0.015g、2水合氯化钙0.015g、1水合硫酸锰0.01g、氯化钴0.02g和硫酸锌0.002g。
发酵培养基：
1.葡萄糖为碳源的培养基：每升培养基中含葡萄糖80g、酵母粉1g、磷酸二氢钾0.5g、磷酸氢二钾0.5g、乙酸铵2.2g、矿物质混合物和维生素。其中，矿物质混合物的组成为：每升培养基中含7水合硫酸镁0.2g、7水合硫酸亚铁0.01g、1水合硫酸锰0.01g和氯化钠0.01g；维生素的组成为：每升培养基中含对氨基苯甲酸0.001g、维生素B1 0.001g和生物素0.00001g。
2.玉米秸秆水解液为碳源的发酵培养基的组成：每升培养基中含葡萄糖48.4g、木糖15.6g、纤维二糖4.8g、阿拉伯糖2.4g、酵母粉1g、磷酸二氢钾0.5g、磷酸氢二钾0.5g、乙酸铵2.2g、7水合硫酸镁0.2g、7水合硫酸亚铁0.01g、1水合硫酸锰0.01g、氯化钠0.01g、对氨基苯甲酸0.001g、维生素B1 0.001g和生物素0.00001g。
3.玉米淀粉为碳源的发酵培养基组成：每升培养基中含还原糖200g、磷酸二氢钾0.5g、磷酸氢二钾0.5g、乙酸铵2.2g、7水合硫酸镁0.2g、7水合硫酸亚铁0.01g、1水合硫酸锰0.01g、氯化钠0.01g、对氨基苯甲酸0.001g、维生素B1 0.001g和生物素0.00001g。发酵初始时玉米淀粉培养基的还原糖浓度可以在60-100g/L，通过加水稀释调配。批式流加过程玉米淀粉发酵培养基中的还原糖浓度可以在150-200g/L.
丙酮丁醇生产菌的培养和发酵：种子培养基在使用之前，通氮气除氧10分钟，然后在121℃灭菌30分钟，冷却到室温后，接入生产菌。将生产菌在种子培养罐1中于37℃的条件下培养15h后，准备接入到发酵培养基中。发酵培养基在使用之前，在121℃灭菌30分钟，然后通入氮气除氧2h，冷却到室温后，通过泵将含有生产菌的种子液(发酵培养基体积的10％)泵入生物反应器中在37℃的条件下开始发酵。发酵培养基初始不调节pH值，当发酵液pH低于5.0后，自动流加氢氧化钠水溶液或氨水，将pH调整到5.0以上。
生物反应器可以为搅拌式生物反应器、气升式生物反应器、或者静置式生物反应器。
气化渗透膜的制备：聚二甲基硅氧烷(PDMS)从购于美国道康宁公司(Dow corning)。沸石纳米材料(ZSM-5)从美国Zeolyst International购买。ZSM-5先在80℃下烘干24小时。聚二甲基硅氧烷(PDMS)中的成胶剂基液和固化剂按10:1的比例混合。对于纯PDMS聚合膜的制备，直接在8000转/分钟下离心5分钟，进行后续操作。对于添加ZSM-5材料的PDMS混合膜，将指定重量比例(20％-80％)的ZSM-5，与按10:1比例混合后的PDMS制膜液混合，在8000转下离心5分钟，进行后续操作。后续操作为先用超声处理制膜液15分钟，除去制膜液中的气泡，然后用刮刀将制膜液均匀涂抹在玻璃板上，将带有制膜液的玻璃板放到100℃的烘箱中3小时成膜。最后从烘箱中取出玻璃板，剥离制备好的渗透气化膜，将其固定在膜分离装置中，用于气化渗透耦合分离纯化操作。
丁醇、丙酮、乙醇的分析使用常规气相色谱法，葡萄糖、还原糖和玉米秸秆水解液中的糖的浓度测定使用常规液相色谱法或DNS法。
比较例1未偶联分离装置的丁醇、丙酮和乙醇发酵(未分离)
按上述方法利用葡萄糖为碳源的发酵培养基进行丙酮丁醇生产菌的培养和发酵。当生产菌接入生物反应器后，开始发酵，直到发酵结束。结果如表1所示，丁醇、丙酮和乙醇在发酵液中的终点浓度为14g/L(1.4％)，7g/L(0.7％)和2g/L(0.2％)左右。
比较例2传统法进行气提耦合发酵生产丁醇、丙酮和乙醇
按上述方法进行丙酮丁醇生产菌的培养和发酵。当生产菌接入生物反应器后，开始发酵。传统法进行气提耦合发酵，发酵开始后，当发酵液中丁醇浓度大于5g/L，启动气提分离装置。传统气提耦合发酵过程，通过向生物反应器鼓泡，使丁醇，丙酮和乙醇汽化。随后，气体和汽化的丁醇，丙酮和乙醇通过冷凝管，使丁醇，丙酮和乙醇以液体形式冷凝回收，冷凝管无纯化浓缩作用。结果如表1所示。气提得到的冷凝液中回收到的丁醇浓度为9.0％-11％(w/v)，丙酮浓度为3.5％-5.0％(w/v)，乙醇浓度为0.5％-1.5％(w/v)。气提回收得到的冷凝液中的丁醇、丙酮和乙醇浓度的变化主要是由发酵液中丁醇、丙酮和乙醇浓度的变化造成。
比较例3传统法进行渗透气化耦合发酵生产丁醇、丙酮和乙醇
按上述方法进行丙酮丁醇生产菌的培养和发酵。当生产菌接入生物反应器后，开始发酵。传统法进行渗透气化耦合发酵，采用发酵开始后，当发酵液中丁醇浓度大于5g/L，启动渗透气化膜分离装置，分离膜采用纯PDMS聚合膜。渗透气化耦合发酵过程使液体形式的发酵液在生物反应器和渗透气化膜分离装置之间循环。当发酵液经过分离膜的一侧时，液体形式的发酵液会以溶解扩散的方式通过分离膜，使发酵液得到浓缩纯化。传统结果如表1所示。结果表明传统的渗透气化分离得到的冷凝液中丁醇、丙酮和乙醇的浓度为7.5％-9.0％(w/v),3.0％-4.0％(w/v)和0.6％-1.0％(w/v)。分离膜如采用PDMS/50％ZSM-5混合膜进行分离纯化，渗透气化分离得到的冷凝液中丁醇、丙酮和乙醇的浓度为14.0％-18.0％(w/v)、6.0％-9.0％(w/v)和1.0％-2.0％(w/v)。渗透气化法回收得到的冷凝液中的丁醇、丙酮和乙醇浓度的变化主要是由发酵液中丁醇、丙酮和乙醇浓度的变化造成。
实施例1以葡萄糖为碳源的气化渗透耦合发酵生产丁醇、丙酮和乙醇
按上述方法进行丙酮丁醇生产菌的培养和发酵。当生产菌接入生物反应器后，开始发酵。当发酵液中的丁醇浓度达到5g/L时，开启生物反应器与膜分离装置之间的泵，向生物反应器内鼓泡，泵使气体和气体夹带的气化状态的丁醇、丙酮和乙醇在生物反应器和膜分离装置所形成的密闭空间循环，当气化状态的丁醇、丙酮和乙醇经过分离膜的一侧时，会溶解扩散到膜的另一侧，被冷凝浓缩回收。当发酵培养基中的葡萄糖浓度降到10g/L以下时，向发酵培养基中补加400g/L的浓缩葡萄糖，继续发酵。由于丁醇、丙酮和乙醇不断地被气化渗透法从生物反应器中回收，发酵培养基中有足够的葡萄糖碳源，发酵可以持续稳定运行150小时以上。采用纯的PDMS聚合膜，气化渗透法得到的冷凝液中丁醇、丙酮和乙醇的浓度为20.0％-24.0％(w/v),8.0％-12.0％(w/v)和0.8％-1.2％(w/v)。采用PDMS/50％ZSM-5混合膜进行分离纯化，气化渗透法得到的冷凝液中丁醇、丙酮和乙醇的浓度为28.0％-32.0％(w/v),11.0％-15.0％(w/v)和1.0％-2.0％(w/v)。可以看出，与比较例2的气提耦合发酵相比，气化渗透法得到的冷凝液中丁醇和丙酮浓度提高了2倍多，乙醇浓度提高了一倍左右。与比较例3渗透气化相比，气化渗透法得到的冷凝液中丁醇和丙酮浓度提高了2倍左右，乙醇浓度几乎没有变化。另外，添加疏水颗粒ZSM-5到PDMS膜中可以提高丁醇和丙酮的分离纯化浓度，提高膜的选择性。添加其他纳米疏水性颗粒到膜中，也可以达到提高丁醇和丙酮分离效率的作用。
实施例2以玉米秸秆水解液为碳源的气化渗透耦合发酵生产丁醇、丙酮和乙醇
按上述方法进行丙酮丁醇生产菌的培养和发酵。当生产菌接入生物反应器后，开始发酵。当发酵液中的丁醇浓度达到5g/L时，开启生物反应器与膜分离装置之间的泵，向生物反应器内鼓泡，泵使气体和气体夹带的气化状态的丁醇、丙酮和乙醇在生物反应器和膜分离装置所形成的密闭空间循环，当气化状态的丁醇、丙酮和乙醇经过分离膜的一侧时，会溶解扩散到膜的另一侧，被冷凝浓缩回收。当发酵培养基中的还原糖浓度降到10g/L以下时，向发酵培养基中补加玉米秸秆水解液，继续发酵。由于丁醇、丙酮和乙醇不断地被气化渗透法从生物反应器中回收，发酵培养基中有足够的碳源补充，发酵可以持续稳定运行。采用纯的PDMS聚合膜，气化渗透法得到的冷凝液中丁醇、丙酮和乙醇的浓度为14.0％-18.0％(w/v),6.0％-9.0％(w/v)和0.6％-1.0％(w/v)。采用PDMS/50％ZSM-5混合膜进行分离纯化，气化渗透法得到的冷凝液中丁醇、丙酮和乙醇的浓度为18.0％-22.0％(w/v),7.0％-11.0％(w/v)和1.0％-1.5％(w/v)。可以看出，与比较例2的气提耦合发酵和比较例3渗透气化耦合发酵相比，气化渗透法得到的冷凝液中丁醇和丙酮浓度都提高很多，说明气化渗透法耦合发酵可以分离纯化得到更高浓度的丁醇和丙酮，对丁醇和丙酮产物有更高的选择性和更好的分离效果。和以葡萄糖为碳源的发酵培养基相比，玉米秸秆水解液中可能存在抑制性产物，对细胞有毒性，影响丁醇和丙酮的生成，发酵液中的丁醇和丙酮的浓度对气化渗透法分离纯化的丁醇和丙酮的浓度有密切影响，即如果发酵液中丁醇和丙酮的浓度低，气化渗透得到的冷凝液中的丁醇浓度也低。
实施例3以玉米淀粉糖化液为碳源的气化渗透耦合发酵生产丁醇、丙酮和乙醇
按上述方法进行丙酮丁醇生产菌的培养和发酵。当生产菌接入生物反应器后，开始发酵。当发酵液中的丁醇浓度达到5g/L时，开启生物反应器与膜分离装置之间的 泵，向生物反应器内鼓泡，泵使气体和气体夹带的气化状态的丁醇、丙酮和乙醇在生物反应器和膜分离装置所形成的密闭空间循环，当气化状态的丁醇、丙酮和乙醇经过分离膜的一侧时，会溶解扩散到膜的另一侧，被冷凝浓缩回收。当发酵培养基中的还原糖浓度降到10g/L以下时，向发酵培养基中补加玉米淀粉糖化液，继续发酵。由于丁醇、丙酮和乙醇不断地被气化渗透法从生物反应器中回收，发酵培养基中有足够的碳源，发酵可以持续稳定运行。采用纯的PDMS聚合膜，气化渗透法得到的冷凝液中丁醇、丙酮和乙醇的浓度为17.0％-22.0％％(w/v),7.0％-11.0％(w/v)和0.7％1.1％(w/v)。采用PDMS/50％ZSM-5混合膜进行分离纯化，气化渗透法得到的冷凝液中丁醇、丙酮和乙醇的浓度为22.0％-30.0％(w/v),9.0％-14.0％(w/v)和1.0％-1.7％(w/v)。可以看出，与比较例2的气提耦合发酵和比较例3渗透气化耦合发酵相比，气化渗透法得到的冷凝液中丁醇和丙酮浓度都提高很多，说明气化渗透法耦合发酵可以分离纯化得到更高浓度的丁醇和丙酮，对丁醇和丙酮产物有更高的选择性和更好的分离效果。
表1
Figure PCTCN2016085818-appb-000001
由上可知，传统不耦合分离的丙酮丁醇发酵，得到的发酵液中丁醇和丙酮浓度分别为1.4％，0.7％和0.2％。传统的一步气提法得到的冷凝液中的丁醇和丙酮浓度分别为9.0％-11.0％,3.5％-5.0％和0.5％-1.5％左右。渗透气化分离得到的冷凝液中丁醇、丙酮和乙醇的浓度为14.0％-18.0％(w/v),6.0％-9.0％(w/v)和1.0％-2.0％(w/v)。本发明利用气化渗透耦合发酵技术最终收集到的产物中含有丁醇、丙酮和乙醇浓度分别为28.0％-32.0％，11.0％-15.0％和1.0％-2.0％左右。可见，气化渗透耦合发酵技术可以得到最高纯化分离的产物浓度。由于最终产物中含有极高浓度的丁醇和丙酮，极易通过简单的脱水处理如精馏，蒸馏或膜分离获得纯丁醇和丙酮。本发明气化渗透过程对于提高丁醇提纯浓度，降低整个发酵分离工艺的能耗至关重要。和传统的精馏分离相比，气化渗透法需要的能量消耗为传统精馏分离的30％左右。并且，保证后续丁醇和丙酮纯化分离可以在富含高浓度丁醇和丙酮的溶液中低能耗，高效率进行。更重要的是，和传统的汽化渗透耦合发酵相比，渗透汽化技术膜的一侧需要发酵液与膜接触， 发酵液中的微生物细胞，糖类物质，蛋白物质和其他大分子物质，在膜长期运行下会对膜造成污染，清洗和更换膜的成本非常高。本发明使用的汽化渗透技术耦合发酵，膜两侧均为汽化状态的有机溶剂，对膜没有任何污染，保证膜的使用寿命。因此，本发明可以提高丁醇和丙酮生产和回收效率并降低分离提纯的能耗，为生物法生产丁醇和丙酮提供新的技术，具有很大的工业应用价值。

Claims (10)

  1. 一种发酵耦合分离纯化丁醇、丙酮和乙醇或纯化丁醇的方法，所述方法包括以下步骤：
    1)利用丙酮丁醇生产菌或丁醇生产菌发酵得到丁醇、丙酮和乙醇或得到丁醇；
    2)采用汽化渗透法从发酵液在线分离纯化丁醇、丙酮和乙醇；
    其中所述汽化渗透法包括以下步骤：
    a.向发酵液中通入气泡，使丁醇、丙酮和乙醇或使丁醇汽化；
    b.使气泡中的气体夹带汽化的丁醇、丙酮和乙醇或汽化的丁醇进入膜分离装置内并透过膜；
    c.回收所述丁醇、丙酮和乙醇或回收所述丁醇，或进入下一级分离装置。
  2. 根据权利要求1所述的方法，其中所述气泡的来源为丙酮丁醇生产菌或丁醇生产菌的自产气体或外源气体，所述自产气体优选为二氧化碳和/或氢气，所述外源气体优选为氮气。
  3. 根据权利要求1所述的方法，其中所述丙酮丁醇生产菌优选选自丙酮丁醇梭菌(Clostridium acetobutylicum)、拜式梭菌(Clostridium beijerinckii)、大肠杆菌或酪丁酸梭菌(Clostridium tyrobutyricum)以及这些菌株的基因工程改造菌，优选为丙酮丁醇梭菌，所述丁醇生产菌为产丁醇的基因工程菌，优选为产丁醇大肠杆菌或产丁醇梭菌。
  4. 根据权利要求1所述的方法，其中所述膜为汽化渗透膜，优选为丁醇和丙酮高选择性有机疏水分离膜或有机无机复合膜，更优选选自硅橡胶、聚三甲基硅丙炔、聚丙烯、聚丁二烯、聚偏氟乙烯、聚四氟乙烯或其衍生物、丁腈橡胶、或分子筛材料中的至少一种，最优选为含PDMS的复合膜，所述膜的类型优选为管式、卷式、平板或中空纤维膜形式。
  5. 根据权利要求1所述的方法，其中所述回收通过冷凝进行，所述冷凝优选用装置冷凝或液氮或冷凝水冷凝，其中所述装置冷凝或冷凝水冷凝的温度为-30-+15℃。
  6. 根据权利要求1所述的方法，其中所述气泡从生物反应器底部通入，通气比为0.5-5vvm。
  7. 根据权利要求1所述的方法，所述膜分离装置的膜透过液一侧保持真空度为0-10kpa，优选<5kpa。
  8. 一种用于发酵耦合分离纯化丁醇、丙酮和乙醇或纯化丁醇的的装置，其包括:
    培养基储罐(1)，用于向生物反应器提供培养基；
    生物反应器(2)，与所述培养基储罐(1)连通，用于发酵；
    气体分散器(9)，用于向发酵液中提供气泡；
    膜分离装置(4)，与所述生物反应器(2)气体连通，接收来自生物反应器的夹带丁醇、丙酮和乙醇或夹带丁醇的气泡并膜分离所述丁醇、丙酮和乙醇或所述丁醇；
    冷凝装置(5)，用于回收所述丁醇、丙酮和乙醇或所述丁醇；
    真空表(6)和真空泵(8)，用于为气态形式的丁醇、丙酮和乙醇或气态形式的丁醇透过提供驱动力；
    产品储罐(7)，用于接收产品。
  9. 根据权利要求8所述的装置，其还包括泵(3)，用于将膜分离装置(4)中的气体输送回所述生物反应器(2)。
  10. 根据权利要求8所述的装置，其中所述膜分离装置(4)中的膜为汽化渗透膜，优选为丁醇和丙酮高选择性有机疏水分离膜或有机无机复合膜，更优选选自硅橡胶、聚三甲基硅丙炔、聚丙烯、聚丁二烯、聚偏氟乙烯、聚四氟乙烯或其衍生物、丁腈橡胶、或分子筛材料中的至少一种，最优选为含PDMS的复合膜，所述膜的类型优选为管式、卷式、平板或中空纤维膜形式。
PCT/CN2016/085818 2016-06-15 2016-06-15 一种发酵耦合分离纯化生产丁醇的方法及装置 Ceased WO2017214873A1 (zh)

Priority Applications (3)

Application Number Priority Date Filing Date Title
US15/737,796 US11118198B2 (en) 2016-06-15 2016-06-15 Method and device for fermentation integrated with separation and purification of alcohols
PCT/CN2016/085818 WO2017214873A1 (zh) 2016-06-15 2016-06-15 一种发酵耦合分离纯化生产丁醇的方法及装置
US17/394,695 US20210363553A1 (en) 2016-06-15 2021-08-05 Device for fermentation integrated with separation and purification of alcohols

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/CN2016/085818 WO2017214873A1 (zh) 2016-06-15 2016-06-15 一种发酵耦合分离纯化生产丁醇的方法及装置

Related Child Applications (2)

Application Number Title Priority Date Filing Date
US15/737,796 A-371-Of-International US11118198B2 (en) 2016-06-15 2016-06-15 Method and device for fermentation integrated with separation and purification of alcohols
US17/394,695 Division US20210363553A1 (en) 2016-06-15 2021-08-05 Device for fermentation integrated with separation and purification of alcohols

Publications (1)

Publication Number Publication Date
WO2017214873A1 true WO2017214873A1 (zh) 2017-12-21

Family

ID=60662928

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2016/085818 Ceased WO2017214873A1 (zh) 2016-06-15 2016-06-15 一种发酵耦合分离纯化生产丁醇的方法及装置

Country Status (2)

Country Link
US (2) US11118198B2 (zh)
WO (1) WO2017214873A1 (zh)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP7838352B2 (ja) * 2021-03-25 2026-04-01 三菱ケミカル株式会社 ガス分離膜、その製造方法、ガス分離膜を用いたモジュール、及びガス分離方法
AU2023447110A1 (en) * 2023-05-08 2025-11-27 Archer-Daniels-Midland Company Method for the recovery of amyl alcohol

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009097322A1 (en) * 2008-01-28 2009-08-06 Promerus Llc Polynorbornene pervaporation membrane films, preparation and use thereof
CN101805754A (zh) * 2010-03-31 2010-08-18 南京工业大学 生物质发酵与渗透汽化耦合原位分离丙酮、丁醇和乙醇的工艺
CN102676589A (zh) * 2012-05-09 2012-09-19 大连理工大学 一种发酵偶联气提的生产和分离纯化丁醇的方法
CN102911854A (zh) * 2012-09-29 2013-02-06 大连理工大学 一种分离纯化丁醇和丙酮的装置及方法

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20130078689A1 (en) * 2011-09-23 2013-03-28 Coskata, Inc. Processes for enhancing the performance of large-scale, stirred tank anaerobic fermentors and apparatus therefor

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009097322A1 (en) * 2008-01-28 2009-08-06 Promerus Llc Polynorbornene pervaporation membrane films, preparation and use thereof
CN101805754A (zh) * 2010-03-31 2010-08-18 南京工业大学 生物质发酵与渗透汽化耦合原位分离丙酮、丁醇和乙醇的工艺
CN102676589A (zh) * 2012-05-09 2012-09-19 大连理工大学 一种发酵偶联气提的生产和分离纯化丁醇的方法
CN102911854A (zh) * 2012-09-29 2013-02-06 大连理工大学 一种分离纯化丁醇和丙酮的装置及方法

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
CHUANG, XUE ET AL.: "A Novel In Situ Gas Stripping-Pervaporation Process Integrated With Acetone-Butanol-Ethanol Fermentation for Hyper n-Butanol Production", BIOTECHNOLOGY AND BIOENGINEERING, vol. 113, 31 January 2016 (2016-01-31), pages 1, XP055447733 *
DI CAI ET AL.: "Gas stripping-pervaporation hybrid process for energy-saving product recovery from acetone-butanol-ethanol (ABE) fermentation broth", CHEMICAL ENGINEERING JOURNAL, vol. 287, 1 March 2016 (2016-03-01), XP029360046 *
LUO, JIANQUAN ET AL.: "Separation and concentration of butanol from acetone-butanol-ethanol mixed solution by pervaporation", CHEMICAL ENGINEERING, vol. 38, no. 2, 15 February 2010 (2010-02-15), XP055447746 *

Also Published As

Publication number Publication date
US20210363553A1 (en) 2021-11-25
US20190093134A1 (en) 2019-03-28
US11118198B2 (en) 2021-09-14

Similar Documents

Publication Publication Date Title
Schügerl Integrated processing of biotechnology products
Wu et al. Extractive fermentation for butyric acid production from glucose by Clostridium tyrobutyricum
Xue et al. Two-stage in situ gas stripping for enhanced butanol fermentation and energy-saving product recovery
Xue et al. Characterization of gas stripping and its integration with acetone–butanol–ethanol fermentation for high-efficient butanol production and recovery
Qureshi et al. Butanol,‘a superior biofuel’production from agricultural residues (renewable biomass): recent progress in technology
CN102676589B (zh) 一种发酵偶联气提的生产和分离纯化丁醇的方法
CN101805754B (zh) 生物质发酵与渗透汽化耦合原位分离丙酮、丁醇和乙醇的工艺
US20140356920A1 (en) Recovery of higher alcohols from dilute aqueous solutions
Xue et al. Evaluation of asymmetric polydimethylsiloxane-polyvinylidene fluoride composite membrane and incorporated with acetone-butanol-ethanol fermentation for butanol recovery
WO2012013004A1 (zh) 盐析萃取发酵液中丙酮和丁醇的方法
Wu et al. Acetone–butanol–ethanol production using pH control strategy and immobilized cells in an integrated fermentation–pervaporation process
CN103695475B (zh) 一种利用膜分离技术原位分离乙醇的方法
US9562242B2 (en) Method for reusing water in fermented butanedioic acid separation process
CN103555560B (zh) 一种丙酮丁醇发酵偶联分离纯化制备丁醇的装置和方法
CN109486876A (zh) 一种发酵、提取和纯化苏氨酸的方法
CN1186450C (zh) 生物质发酵与膜渗透汽化制备无水乙醇的方法
CN104877889A (zh) 发酵耦合蒸汽渗透原位分离挥发性有机物的方法及装置
JP2010161987A (ja) 発酵によるブタノールの製造方法
US20210363553A1 (en) Device for fermentation integrated with separation and purification of alcohols
CN102911854B (zh) 一种分离纯化丁醇和丙酮的装置及方法
CN102492750A (zh) 利用赤霉菌ca3-1转化3-氰基吡啶为烟酸的方法
Choudhury et al. Lactic acid fermentation in cell-recycle membrane bioreactor
RU2644239C2 (ru) Способ продуцирования с2-оксигенатов путем ферментации с использованием серы с высокой степенью окисления
JP2003284580A (ja) 乳酸エステルの製造方法および装置
CN103468606B (zh) 一株产酸克雷伯氏菌及其在生产蒜糖醇中的应用

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 16904990

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 16904990

Country of ref document: EP

Kind code of ref document: A1