CN104130934A - Immobilized fibrous-bed reactor for producing 1,3-propylene glycol by fermentation - Google Patents
Immobilized fibrous-bed reactor for producing 1,3-propylene glycol by fermentation Download PDFInfo
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Abstract
The invention provides an immobilized fibrous-bed reactor for producing 1,3-propylene glycol by fermentation, and relates to 1,3-propylene glycol. The immobilized fibrous-bed reactor is provided with a reactor body, a fermentation tank and an automatic pH control device, wherein the automatic pH control device is provided with a fermentation tank pH probe, a reactor pH probe, a fermentation tank pH detector, a reactor pH detector, alkali liquor peristaltic pumps, alkali liquor bottles and air filters; an alkali liquor outlet of the first alkali liquor bottle is connected to the fermentation tank by virtue of the first alkali liquor peristaltic pump; the fermentation tank pH probe is connected with the fermentation tank, the output end of the fermentation tank pH probe is connected with the fermentation tank pH detector, and an outlet of the fermentation tank air filter is connected with the fermentation tank; an outlet of the fermentation tank is connected with the reactor by virtue of a fermentation broth peristaltic pump; an outlet of the second alkali liquor bottle is connected with the reactor by virtue of a peristaltic pump, and the air filter is arranged on the second alkali liquor bottle; the reactor pH probe is connected with the reactor, and a fiber material is filled in the reactor; the reactor pH probe is connected with the reactor pH detector; and a sieve plate is arranged at the bottom in the reactor; a tail gas outlet is formed in the reactor, and the reactor is connected with a product collection device.
Description
Technical field
The present invention relates to the production unit of 1,3-PD, relate in particular to a kind of immobilization fibre bed reactor of producing 1,3-propanediol through fermentation.
Background technology
1,3-PD (1,3-propanediol, write a Chinese character in simplified form: 1,3-PD) be a kind of important industrial chemicals.Because it contains bifunctional group, can participate in number of chemical building-up reactions, for example, as monomer, can produce the polycondensates such as polyester that special occasions uses, polyethers, urethane; As the raw material of organic synthesis intermediate, can be used for the synthetic of multi-medicament and novel antioxidant; As solvent, can improve premium properties (the Kaur G of the products such as tamanori, resin, washing composition, makeup, Srivastava A K, Chand S.Advances in biotechnological production of 1,3-propanediol[J] .Biochemical Engineering Journal, 2012,64:106-118).In addition, with 1,3-PD, replace BDO, also can be used for producing thermoplastic polyurethane, copolyester, engineering plastics etc.1, ammediol dehydration and dehydrogenation can production and tetrahydrofuran (THF) and the similar products of gamma-butyrolactone chemical property, as trimethylene oxide, and by ring-opening polymerization, produce the new polymers that is similar to polytetramethylene ether diol (PTMEG), or as the reaction solvent in coating.1, ammediol and terephthalic acid can synthesize polytrimethylene terephthalate (PTT), make 1, ammediol extremely common people is paid close attention to (Deckwer W D.Microbial conversion of glycerol to 1,3-propanediol[J] .FEMS Microbiology Reviews, 1995,16:143-149).This is due to the chemical resistant properties of the existing polyester of PTT, to have again good rebound resilience, pollution resistance, the anti-fold of tynex, and be easy to dyeing, soft, high resilience, the easy characteristic such as dry, low static behaviour, biological degradability, can be used for the industries such as carpet, engineering plastics, film and dress materials, be regarded as the another new type polyester fiber after polyethylene terephthalate (PET) and polybutylene terephthalate (PBT), there are wide market outlook.
With the synthetic 1,3-PD of chemical method, not only production cost is high but also contaminate environment, the at present domestic chemical method industrialization report that there is no.Biofuel is as reproducible clean energy, in its production process, and the highest approximately 10% the by-product glycerin that obtains; As the 1,3-PD of important industrial chemicals, caused people's extensive concern; So as take biological diesel oil byproduct glycerin as raw material production 1,3-PD, not only solved the problem of outlet of biological diesel oil byproduct glycerin, and reduced the production cost of 1,3-PD.Thereby adopt 1,3-propanediol biosynthesis to there are huge potentiality.
Immobilized cell bioreactor has that cell density is high, catalytic efficiency is high, resistance to murder by poisoning ability is strong, product separation easily, can realize operate continuously, be easy to amplify and control reaction process, the advantages such as downstream processing is easy are in current continuous mass production, to apply maximum a kind of bio-reactors.Set up low, the safe 1,3-PD production technique of cost, for comprehensive utilization biological diesel oil byproduct glycerin provides effective reference frame, to obtaining higher economic worth and social value.
The citrobacter freundii (Citrobacter freundii) of take is example, the people such as U.Pflugmacher (U.Pflugmacher, G.Gottschalk.Development of an immobilized cell reactor for the production of 1,3-propanediol by Citrobacter freundii[J] .Appl.Microbiol Biotechnol.1994,41:313-316) use polyurethane material as fixation support, reactor is the anaerobic fixbed reactor with return-flow system and pH regulator system.As 37 ℃ of temperature, pH6.9, thinning ratio 0.5h
-1, during feed supplement liquid glycerol concentration 0.4mol/L, the 1,3-PD largest production intensity detecting is 8.2g/L/h.The Cray Bai Shi pneumobacillus (Klebsiella pneumoniae) of take is example, (the Y.N.Zhao such as Ya-Nan Zhao, G.Chen, S.J.Yao.Microbial production of 1,3-propanediol from glycerol by encapsulated Klebsiella pneumoniae[J] .Biochemical Engineering Journal.2006,32:93-99) that Cray Bai Shi pneumobacillus is microcapsule embedded with NaCS/PDMDAAC, utilize glycerol production 1,3-PD.In microcapsule, cell concentration can reach 6.84g/L, and specific ionization state is high 2.6 times, has effectively improved substrate tolerance and cell stability.In batch fermentation, 1,3-PD concentration can reach 63.1g/L, and transformation efficiency is 0.65mol1, ammediol/mol glycerine.After batch fermentation 67h, microcapsule immobilized cell still keeps certain activity, and 1,3-PD concentration is 51.86g/L.In thinning ratio, be 0.33h
-1continuously ferment, 1,3-PD concentration is 13.6g/L, production intensity is 4.49g/L/h.Traditional immobilization fermentation exists the shortcoming that shorter, product production intensity of cycle sharply declines after fermentation for some time, this be mainly due to the accumulation of dead cell on fixation support and high density fermentation later stage mass transfer poor.Microcapsule fixation method is because cost is high, and is subject to mass transfer condition restriction, and cell is difficult to realize self to guarantee for a long time thalline vigor, is not suitable for producing continuously extensive, for a long time.
Fibre bed bio-reactor (fibrous bed bioreactor, FBB) people such as professor Yang Shangtian of Shi You Ohio State Univ-Columbus USA (Yang S T, White S A, Hsu S T.Extraction of carboxylic acids with tertiary and quaternary amines:effect of pH[J] .Industrial & engineering chemistry research, 1991, 30 (6): 1335-1342) at traditional packed bed reactor (packed bed reactor, that PBR) on the basis of performance, designs is a kind of simple, efficient bio-reactor, and be first applied to propionic fermentation.The filamentary materials such as cotton fabric, as the carrier of fixed cell, have porosity high (>95%), specific surface area (>40m
2/ m
3) advantage such as large, carrier active high, cost is low, physical strength is high, unreactiveness.Immobilized cell in FBB can be constantly updated, and dead cell can along with flowing of substratum, Automatic-falling adheres to space to be substituted by neonatal cell, keeps higher viable cell ratio on fixation support always., owing to having adopted the filamentary material of high porosity and high-specific surface area as fixation support, compare with traditional immobilization way, the mass-transfer efficiency of FBB is higher meanwhile.FBB can move 6 months to 1 year continuously, also can effectively avoid the FAQs in the immobilization fermentations such as pollution and obstruction simultaneously.Utilize at present fibre bed reactor to produce 1,3-PD and have no report.
The applicant discloses a kind of bacterial strain screening method and application of producing 1,3-PD in Chinese patent CN102965323A, relates to 1,3-PD.The bacterial strain screening method and the application that clostridium butylicum (Clostridium butyricum) Gen160 are provided and utilize useless glycerol production 1,3-PD.Step joins in cabbage juice for getting collected specimens, the rearmounted water-bath of enrichment culture, kill non-sporeformer, proceed to clostridium propagation liquid nutrient medium, anaerobism is cultivated and is put water-bath again, proceeds to fermention medium, carries out efficient liquid phase chromatographic analysis tunning after anaerobic selection enrichment culture, choose the positive bacteria that can produce 1,3-PD; Adopt deep agar post method to isolate single bacterium colony; Carry out aerobic and anaerobism and cultivate, remove facultative anaerobe, obtain strictly anaerobic bacterium, analyze tunning, choose and can produce the bacterial strain that 1,3-PD and cultural characteristic, colonial morphology all meet clostridium butylicum cultural characteristic and carry out 16SrDNA sequential analysis evaluation.Described bacterial classification is clostridium butylicum (Clostridium butyricum) Gen160, this bacterial classification has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on 07 05th, 2012, deposit number is CGMCCNo.6317, address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode 100101.
Summary of the invention
The object of the invention is to the deficiency existing for above-mentioned prior art, a kind of immobilization fibre bed reactor of producing 1,3-propanediol through fermentation is provided.
The present invention is provided with reactor, fermentor tank, pH automatic control device;
Described pH automatic control device is provided with fermentor tank pH probe, reactor pH probe, fermentor tank pH detector, reactor pH detector, the 1st alkali lye peristaltic pump, the 2nd alkali lye peristaltic pump, the 1st alkali lye bottle, the 2nd alkali lye bottle, the 1st air filter, fermentor tank air filter, the 2nd air filter and connecting pipeline, air is in the 1st air filter access the 1st alkali lye bottle, and the alkali lye outlet of the 1st alkali lye bottle is in the 1st alkali lye peristaltic pump access fermentor tank; Fermentor tank pH probe is connected in fermentor tank, and the output terminal of fermentor tank pH probe connects fermentor tank pH detector, in the delivery port access fermentor tank of fermentor tank air filter; On fermentor tank, be connected to fermentor tank gas inlet; The fermented liquid outlet of fermentor tank connects reactor through fermented liquid peristaltic pump; The alkali lye outlet of the 2nd alkali lye bottle is in the 2nd alkali lye peristaltic pump access reactor, and the 2nd alkali lye bottle is provided with the 2nd air filter; Reactor pH probe is connected in reactor, is filled with filamentary material in reactor, and reactor pH probe is connected with reactor pH detector, and reactor inner bottom part is provided with porous sieve plate; Reactor is provided with reactor tail gas outlet, and reactor is connected with product collection device by pipeline.
Described product collection device can adopt product collection bottle, can be provided with tail gas outlet on product collection bottle.
Described filamentary material can adopt a kind of in vegetable fibre, animal fibre, synthon etc., and described vegetable fibre can be selected from gauze, cotton etc.; Described animal fibre can be selected from wool fabric etc.; Described synthon can be selected from polyester piece good etc.Filamentary material can be coiled into cylindric being fixed in reactor, as cell immobilization carrier.
When pH is lower than controlled pH in container, by peristaltic pump, the alkali lye in alkali lye bottle is filled into the automatic pH of control in fermentor tank and reactor; Feed supplementing device fills into reactor by the substratum in fermentor tank by the pipeline being connected on reactor by peristaltic pump, and reactor top arranges a tail gas by pipeline and exports.
The present invention can be used for the production of 1,3-PD, and the method that employing the present invention produces 1,3-PD is as follows:
By 1, the ammediol fermentation thalline seed liquor by volume inoculum size of per-cent 4%～10% is inoculated in the interior fermention medium of fermentor tank, from gas inlet, pass into rare gas element, cultivate 6～12h, then by fermentor tank and reactors in series 1～3h, realize the absorption (being that thalline is fixed on filamentary material) of thalline; By feed supplementing device, fermention medium is filled in reactor, from gas inlet, pass into rare gas element, utilize pH automatic control device to fill into alkali lye, control the fermented liquid pH value in reactor, the immobilization that realizes 1,3-PD production bacterium is continuously fermented.
Described 1, ammediol fermentation thalline is clostridium butylicum (Clostridium butyricum) Gen160, this bacterial classification has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on 07 05th, 2012 by the applicant, deposit number is that (address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City to CGMCC No.6317, Institute of Microorganism, Academia Sinica, postcode 100101), referring to the applicant's disclosed information in Chinese patent CN102965323A.
Described rare gas element can adopt nitrogen; Tail gas is the gas mixture of nitrogen, hydrogen and carbonic acid gas.
Described seed liquor is the seed culture medium of having inoculated bacterial strain.
Described fermented liquid is the fermention medium of having inoculated bacterial strain.
Described seed culture medium is (/L): glycerine, 10～40g; K
2hPO
43H
2o, 2～5g; KH
2pO
4, 1～3g; (NH
4)
2sO
4, 2～5g; MgSO
47H
2o, 2～5g; CaCl
22H
2o, 2～5g; Yeast powder, 1～10g; Trace element solution, 1～10mL; Fe solution, 1～10mL; Nature pH.
Described fermention medium is (/L): glycerine (glycerol content is measured by liquid-phase chromatographic analysis) 30～500g/L; K
2hPO
43H
2o, 0.1～2.0g; KH
2pO
4, 0.1～2.0g; (NH
4)
2sO
4, 0.5～5.0g; MgSO
47H
2o, 0.1～1.0g; CaCl
22H
2o, 0.01～0.2g; Yeast powder, 0.5～5.0g; Trace element solution, 1～10mL; Fe solution 1～10mL.
Described Fe solution is (/L): FeSO
47H
2o, 1～10g; HCl (37%), 1～40mL.
Described trace element solution is (/L): ZnCl2,0.01～0.2g; MnCl
24H
2o, 0.01～0.2g; H
3bO
3, 0.01～0.1g; CoCl
26H
2o, 0.1～0.5g; CuCl
22H
2o, 0.01～0.05g; NiCl
26H
2o, 0.02～0.05g; Na
2moO
42H
2o, 0.01～0.06g; HCl (37%), 1～9mL.
Described alkali lye is the sodium hydroxide solution of volumetric molar concentration 1～15mol/L.
Compare with other process for fixation, advantage of the present invention is:
1. device is simplified: the fibre bed reactor device that the present invention builds does not need to utilize immobilization post to realize cell fixing on filamentary material, but filamentary material is fixed in reactor, directly realize the immobilization of cell, saved immobilization post, improved the utilization ratio of reactor, device is simplified.
2. the efficient adsorption of cell and continuous renewal: the filamentary material in fibre bed reactor utilizes the reticulated structure of self, can guarantee the efficient adsorption of some amount thalline.Meanwhile, different from other process for fixation, only there is high-efficiency activated cell and just can be adsorbed in fibre bed, the cell meeting Automatic-falling in the paracme, has guaranteed the vigor of cell in fibre bed.
3. mass-transfer efficiency improves: different from entrapping method, the fibre bed reactor of structure directly contacts with fermented liquid, has removed material transfer process from, has improved mass-transfer efficiency.
4. cell " domestication ": utilize immobilization fibre bed reactor can " tame " cell, the specific culture environment of reactor changes and is applicable to new growing environment making cell self that some occur, through immobilization fibre bed reactor " domestication ", the bacterial strain filtering out will have stronger adaptive faculty and throughput.
5. cell fixation cost is lower: compare with entrapping method, filamentary material cost used in fibre bed reactor is low, durable in use, is applicable to the fermentative production of 1,3-PD.
Accompanying drawing explanation
Fig. 1 is the structural representation of the embodiment of the present invention.In figure, be respectively labeled as: 1-1. the 1st air filter; 1-2. fermentor tank air filter; 1-3. the 2nd air filter; The outlet of 1-4. reactor tail gas; The outlet of 1-5. product bottle tail gas; 2-1. the 1st alkali lye bottle; 2-2. the 2nd alkali lye bottle; 3-1. the 1st alkali lye peristaltic pump; 3-2. fermented liquid peristaltic pump; 3-3. the 2nd alkali lye peristaltic pump; 4-1. fermentor tank pH detector; 4-2. reactor pH detector; 5. fermentor tank gas inlet; 6. fermentor tank; 7-1. fermentor tank pH probe; 7-2. reactor pH probe; 8. reactor; 9. filamentary material; 10. porous sieve plate; 11. product collection bottles.
Fig. 2 is 1,3-PD efficient liquid phase chromatographic analysis figure.In figure: 1,3-PD appearance time is 20.103min.
Embodiment
Following examples are to further illustrate of the present invention.
According to Fig. 1, the embodiment of the present invention is provided with reactor 8, fermentor tank 6, pH automatic control device.
Described pH automatic control device is provided with fermentor tank pH probe 7-1, reactor pH probe 7-2, fermentor tank pH detector 4-1, reactor pH detector 4-2, the 1st alkali lye peristaltic pump 3-1, the 2nd alkali lye peristaltic pump 3-3, the 1st alkali lye bottle 2-1, the 2nd alkali lye bottle 2-2, the 1st air filter 1-1, fermentor tank air filter 1-2, the 2nd air filter 1-3 and connecting pipeline, air is in the 1st air filter 1-1 access the 1st alkali lye bottle 2-1, and the alkali lye outlet of the 1st alkali lye bottle 2-1 is in the 1st alkali lye peristaltic pump 3-1 access fermentor tank 6; Fermentor tank pH probe 7-1 is connected in fermentor tank 6, and the output terminal of fermentor tank pH probe 7-1 meets fermentor tank pH detector 4-1, in the delivery port access fermentor tank 6 of fermentor tank air filter 1-2; On fermentor tank 6, be connected to fermentor tank gas inlet 5; The fermented liquid outlet of fermentor tank 6 connects reactor 8 through fermented liquid peristaltic pump 3-2; The alkali lye outlet of the 2nd alkali lye bottle 2-2 is in the 2nd alkali lye peristaltic pump 3-3 access reactor 8, and the 2nd alkali lye bottle 2-2 is provided with the 2nd air filter 1-3; Reactor pH probe 7-2 is connected in reactor 8, is filled with filamentary material 9 in reactor 8, and reactor pH probe 7-2 is connected with reactor pH detector 4-2, and reactor 8 inner bottom parts are provided with porous sieve plate 10; Reactor 8 is provided with reactor tail gas outlet 1-4, and reactor 8 is connected with product collection device by pipeline.
Described product collection device can adopt product collection bottle 11, and product collection bottle 11 is provided with tail gas outlet 1-5.
Described filamentary material can adopt a kind of in vegetable fibre, animal fibre, synthon etc., and described vegetable fibre can be selected from gauze, cotton etc.; Described animal fibre can be selected from wool fabric etc.; Described synthon can be selected from polyester piece good etc.Filamentary material can be coiled into cylindric being fixed in reactor, as cell immobilization carrier.
When pH is lower than controlled pH in container, by peristaltic pump, the alkali lye in alkali lye bottle is filled into the automatic pH of control in fermentor tank and reactor; Feed supplementing device fills into reactor by the substratum in fermentor tank by the pipeline being connected on reactor by peristaltic pump, and reactor top arranges a tail gas by pipeline and exports.
The present invention can be used for the production of 1,3-PD, and the method that employing the present invention produces 1,3-PD is as follows:
By 1, the ammediol fermentation thalline seed liquor by volume inoculum size of per-cent 4%～10% is inoculated in the interior fermention medium of fermentor tank, from gas inlet, pass into rare gas element, cultivate 6～12h, then by fermentor tank and reactors in series 1～3h, realize the absorption (being that thalline is fixed on filamentary material) of thalline; By feed supplementing device, fermention medium is filled in reactor, from gas inlet, pass into rare gas element, utilize pH automatic control device to fill into alkali lye, control the fermented liquid pH value in reactor, the immobilization that realizes 1,3-PD production bacterium is continuously fermented.
Described 1, ammediol fermentation thalline is clostridium butylicum (Clostridium butyricum) Gen160, this bacterial classification has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on 07 05th, 2012 by the applicant, deposit number is that (address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City to CGMCC No.6317, Institute of Microorganism, Academia Sinica, postcode 100101), referring to the applicant's disclosed information in Chinese patent CN102965323A.
Described rare gas element can adopt nitrogen; Tail gas is the gas mixture of nitrogen, hydrogen and carbonic acid gas.
Described seed liquor is the seed culture medium of having inoculated bacterial strain.
Described fermented liquid is the fermention medium of having inoculated bacterial strain.
Described seed culture medium is (/L): glycerine, 10～40g; K
2hPO
43H
2o, 2～5g; KH
2pO
4, 1～3g; (NH
4)
2sO
4, 2～5g; MgSO
47H
2o, 2～5g; CaCl
22H
2o, 2～5g; Yeast powder, 1～10g; Trace element solution, 1～10mL; Fe solution, 1～10mL; Nature pH.
Described fermention medium is (/L): glycerine (glycerol content is measured by liquid-phase chromatographic analysis) 30～500g/L; K
2hPO
43H
2o, 0.1～2.0g; KH
2pO
4, 0.1～2.0g; (NH
4)
2sO
4, 0.5～5.0g; MgSO
47H
2o, 0.1～1.0g; CaCl
22H
2o, 0.01～0.2g; Yeast powder, 0.5～5.0g; Trace element solution, 1～10mL; Fe solution 1～10mL.
Described Fe solution is (/L): FeSO
47H
2o, 1～10g; HCl (37%), 1～40mL.
Described trace element solution is (/L): ZnCl
2, 0.01～0.2g; MnCl
24H
2o, 0.01～0.2g; H
3bO
3, 0.01～0.1g; CoCl
26H
2o, 0.1～0.5g; CuCl
22H
2o, 0.01～0.05g; NiCl
26H
2o, 0.02～0.05g; Na
2moO
42H
2o, 0.01～0.06g; HCl (37%), 1～9mL.
Described alkali lye is the sodium hydroxide solution of volumetric molar concentration 1～15mol/L.
Filamentary material (40 * 40cm) is coiled into cylindric being fixed in reactor (internal diameter 4cm, effecting reaction height 40cm) as cell immobilization carrier, and pH automatic control device is connected with reactor with feed supplementing device.On fermentor tank, be connected to gas inlet, pH automatic control system, pneumatic outlet and interior agitating device and fermented liquid outlet pump.Feed supplementing device fills into reactor 8 by the substratum in fermentor tank 6 by the pipeline being connected on reactor 8 by fermented liquid peristaltic pump 3-2.Reactor 8 tops arrange a reactor tail gas outlet 1-4 by pipeline.
Below illustrate implementing procedure.
Embodiment 1:
Utilize clostridium butylicum Gen 160 in immobilization fibre bed reactor, to produce 1,3-PD.
Seed culture medium (gL
-1): glycerine, 20; K
2hPO
43H
2o, 4.45; KH
2pO
4, 1.3; (NH
4)
2sO
4, 2; MgSO
47H
2o, 0.2; CaCl
22H
2o, 0.02; Yeast powder, 1.Trace element solution 1mL, Fe solution 1mL.
Fermention medium (gL
-1): glycerine, 50; K
2hPO
43H
2o, 1; KH
2pO
4, 0.5; (NH
4)
2sO
4, 2; MgSO
47H
2o, 0.2; CaCl
22H
2o, 0.02; Yeast powder, 1.Trace element solution 1mL; Fe solution 1mL; PH6.5.
Fe solution: FeSO
47H
2o, 5gL
-1; HCl (37%), 4mlL
-1.
Trace element solution (gL
-1): ZnCl
2, 0.07; MnCl
24H
2o, 0.1; H
3bO
3, 0.06; CoCl
26H
2o, 0.2; CuCl
22H
2o, 0.02; NiCl
26H
2o, 0.025; Na
2moO
42H
2o, 0.035; HCl (37%), 0.9mLL
-1.
By the inoculum size of 10% (v/v), clostridium butylicum Gen 160 seed liquor accesses are equipped with in the fermentor tank of 2L fermention medium, 37 ℃ of constant temperature, rotating speed 150rpm, control pH7.0, passes into 0.1L/min N from gas inlet
2, 8h is to logarithmic phase in fermentation.Reactor passes into N in advance
2deoxygenation, after 8h, with peristaltic pump, by the fermented liquid in fermentor tank, the flow velocity with 20mL/min fills in reactor, and thalline is fixed on cotton fibre material.In fermentor tank, newly add 2L fermention medium, use peristaltic pump with thinning ratio 0.33h
-1to reactor, filling into fermention medium, fill into 4mol/L NaOH solution control pH7.0 in reactor with peristaltic pump, is 37 ℃ by regulating chuck thermostat(t)ed water to control temperature of reactor.After 35h, 1,3-PD concentration can reach 15g/L, production intensity 5.91g/L/h.
Embodiment 2:
Utilize clostridium butylicum Gen 160 in immobilization fibre bed reactor, to produce 1,3-PD.
Substratum is with embodiment 1.
By the inoculum size of 10% (v/v), clostridium butylicum Gen 160 seed liquor accesses are equipped with in the fermentor tank of 2L fermention medium, 37 ℃ of constant temperature, rotating speed 150rpm, control pH7.0, passes into 0.1L/min N from gas inlet
2, 8h is to logarithmic phase in fermentation.4mol/L NaOH solution is housed in alkali lye bottle.Reactor passes into N in advance
2deoxygenation, after 8h, with peristaltic pump, by the fermented liquid in fermentor tank, the flow velocity with 40mL/min fills in reactor, and thalline is fixed on composite fibre materials.In fermentor tank, newly add the fermention medium that 2L is 800g containing amounts of glycerol, use peristaltic pump with thinning ratio 0.1h
-1to reactor, filling into fermention medium, fill into 4mol/L NaOH solution control pH7.0 in reactor with peristaltic pump, is 35 ℃ by regulating chuck thermostat(t)ed water to control temperature of reactor.After 3d, 1,3-PD concentration can reach 65g/L, production intensity 1.01g/L/h.
The present invention can be provided with temperature-control device, and the chuck layer that temperature-control device passes into reactor periphery by thermostat(t)ed water is controlled in reactor in steady temperature and fermentor tank steady temperature in temperature controlling system controlled fermentation tank.
Claims (7)
1. an immobilization fibre bed reactor for producing 1,3-propanediol through fermentation, is characterized in that being provided with reactor, fermentor tank, pH automatic control device;
Described pH automatic control device is provided with fermentor tank pH probe, reactor pH probe, fermentor tank pH detector, reactor pH detector, the 1st alkali lye peristaltic pump, the 2nd alkali lye peristaltic pump, the 1st alkali lye bottle, the 2nd alkali lye bottle, the 1st air filter, fermentor tank air filter, the 2nd air filter and connecting pipeline, air is in the 1st air filter access the 1st alkali lye bottle, and the alkali lye outlet of the 1st alkali lye bottle is in the 1st alkali lye peristaltic pump access fermentor tank; Fermentor tank pH probe is connected in fermentor tank, and the output terminal of fermentor tank pH probe connects fermentor tank pH detector, in the delivery port access fermentor tank of fermentor tank air filter; On fermentor tank, be connected to fermentor tank gas inlet; The fermented liquid outlet of fermentor tank connects reactor through fermented liquid peristaltic pump; The alkali lye outlet of the 2nd alkali lye bottle is in the 2nd alkali lye peristaltic pump access reactor, and the 2nd alkali lye bottle is provided with the 2nd air filter; Reactor pH probe is connected in reactor, is filled with filamentary material in reactor, and reactor pH probe is connected with reactor pH detector, and reactor inner bottom part is provided with porous sieve plate; Reactor is provided with reactor tail gas outlet, and reactor is connected with product collection device by pipeline.
2. a kind of immobilization fibre bed reactor of producing 1,3-propanediol through fermentation as claimed in claim 1, is characterized in that described product collection device adopts product collection bottle, and product collection bottle is provided with tail gas outlet.
3. a kind of immobilization fibre bed reactor of producing 1,3-propanediol through fermentation as claimed in claim 1, is characterized in that described filamentary material adopts a kind of in vegetable fibre, animal fibre, synthon.
4. a kind of immobilization fibre bed reactor of producing 1,3-propanediol through fermentation as claimed in claim 3, is characterized in that described vegetable fibre is selected from gauze, cotton.
5. a kind of immobilization fibre bed reactor of producing 1,3-propanediol through fermentation as claimed in claim 3, is characterized in that described animal fibre is selected from wool fabric.
6. a kind of immobilization fibre bed reactor of producing 1,3-propanediol through fermentation as claimed in claim 3, is characterized in that described synthon are selected from polyester piece good.
7. a kind of immobilization fibre bed reactor of producing 1,3-propanediol through fermentation as claimed in claim 1, is characterized in that filamentary material is coiled into cylindric being fixed in reactor, as cell immobilization carrier.
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| CN114921376A (en) * | 2022-06-01 | 2022-08-19 | 浙江省农业科学院 | High-density culture device and method for clostridium butyricum |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101182455A (en) * | 2007-11-29 | 2008-05-21 | 南京工业大学 | Device and technique for producing propanoic acid by coupling of fibrous bed bioreactor with extraction separation |
| KR20110032482A (en) * | 2009-09-23 | 2011-03-30 | 한국과학기술연구원 | Method and apparatus for production of 1,3-propanediol using glycerol |
| CN103555776A (en) * | 2013-10-31 | 2014-02-05 | 南京工业大学 | Method for producing D-lactic acid through repeated batch fermentation by utilizing surface immobilization technology |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101182455A (en) * | 2007-11-29 | 2008-05-21 | 南京工业大学 | Device and technique for producing propanoic acid by coupling of fibrous bed bioreactor with extraction separation |
| KR20110032482A (en) * | 2009-09-23 | 2011-03-30 | 한국과학기술연구원 | Method and apparatus for production of 1,3-propanediol using glycerol |
| CN103555776A (en) * | 2013-10-31 | 2014-02-05 | 南京工业大学 | Method for producing D-lactic acid through repeated batch fermentation by utilizing surface immobilization technology |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114921376A (en) * | 2022-06-01 | 2022-08-19 | 浙江省农业科学院 | High-density culture device and method for clostridium butyricum |
| CN114921376B (en) * | 2022-06-01 | 2024-03-19 | 浙江省农业科学院 | Clostridium butyricum high-density culture device and method |
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