CN104178421B - A kind of is the method for fluidized bed type enzyme reactor by stirred-tank fermenter system assembles - Google Patents
A kind of is the method for fluidized bed type enzyme reactor by stirred-tank fermenter system assembles Download PDFInfo
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- CN104178421B CN104178421B CN201410420193.6A CN201410420193A CN104178421B CN 104178421 B CN104178421 B CN 104178421B CN 201410420193 A CN201410420193 A CN 201410420193A CN 104178421 B CN104178421 B CN 104178421B
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M21/00—Bioreactors or fermenters specially adapted for specific uses
- C12M21/18—Apparatus specially designed for the use of free, immobilized or carrier-bound enzymes
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
- C12M25/16—Particles; Beads; Granular material; Encapsulation
- C12M25/20—Fluidized bed
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
- C12M41/18—Heat exchange systems, e.g. heat jackets or outer envelopes
- C12M41/22—Heat exchange systems, e.g. heat jackets or outer envelopes in contact with the bioreactor walls
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/26—Means for regulation, monitoring, measurement or control, e.g. flow regulation of pH
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Abstract
The invention provides a kind of method stirring-type biological fermentation tank being assembled into fluidized bed type enzyme reactor, belong to bioreactor design and renovation technique field.The jacket water (J.W.) of external for the water-bath system of stirring-type biological fermentation tank sandwich chromatography column is imported and exported, utilize the peristaltic pump on fermenter system fast by substrate solution from below mouth press-in be equipped with in the chromatography column of immobilized enzyme, immobilized enzyme is suspended, fully contacts with substrate solution.This invention utilizes stirred-tank fermenter device systems to be assembled into fluidized bed type enzyme reactor, realizes the enzymic catalytic reaction of temperature automatically controlled control pH, saves equipment funds and reduce place occupancy volume.
Description
Technical field
The invention belongs to the bioreactor design in bioengineering field and renovation technique field, relating to a kind of is the method for fluidized bed type enzyme reactor by stirred-tank fermenter rapid-assembling.
Background technology
Enzyme especially immobilized enzyme, when in vitro carries out catalyzed reaction, all must carry out in certain reaction vessel, to control various condition and the speed of enzymic catalytic reaction.Enzyme reactor is the core apparatus for completing enzymatic reaction, and it provides suitable place and best reaction conditions for enzymic catalytic reaction, and it is in the Central Position of enzyme-catalyzed reaction, is the bridge connecting raw material and product.
The feature of fluidized bed reactor be substrate solution with enough large flow velocity, from reactor bottom upwards by immobilized enzyme column bed time, immobilized enzyme particle just can be made to be in fluidized state all the time.Fluidized bed reactor can be used for processing the substrate solubility of viscosity comparatively greatly with containing solid particulate, also can be used for the enzyme reaction needing supplied gas or emission gases, its flow pattern makes the degree of mixing of reaction solution mix between type and the horizontal sliding flow pattern of filling bed type enzyme reactor between the complete of continuously stirring formula enzyme reactor.
The enzyme huge number used in industrial enzyme catalytic reaction process, enzyme catalyst form (immobilized enzyme, resolvase, immobilized cell, free cell etc.) is various, substrate physical chemical differences is large, Dynamics of Enzyme Catalysis character and enzyme stability all there are differences, all of these factors taken together all requires to design the enzyme reactor adapting to catalytic process and require, but commercialization enzyme reactor, based on stirred tank, is difficult to the requirement adapting to fast-developing enzyme engineering technology at present.The invention provides one on fermentor tank original system basis, rapid-assembling is the method for fluidized bed type enzyme reactor, realizes the equipment integration of fermentor tank and fluidized-bed reactor, saves facility investment.
Summary of the invention
The object of the invention is to provide a kind of method stirring-type biological fermentation tank being assembled into fluidized bed type enzyme reactor; retain biological fermentation tank original function simultaneously; realize a kind of equipment and have two kinds of (fermentor tank, fluidized bed type enzyme reactor) using function, proper scale is promoted the use of.
The present invention can be reached by following measure: a kind of is the method for fluidized bed type enzyme reactor by stirred-tank fermenter system assembles, it is characterized in that comprising following steps:
1. stirred-tank fermenter system: fermentor tank main body, Controlling System, water bath circulation device, vapour generator, air compressor pump and air accumulator all possess.
2. imported and exported with the chuck of the external chromatography column of silica gel hose respectively by the intake-outlet of the water bath circulation device of stirred-tank fermenter, all there is core at chromatography column upper and lower square opening 10cm place, and wherein going up square opening is that movable core adds to facilitate immobilized enzyme.Add immobilized enzyme by square opening on chromatography column in quiet heavy mode, and by damping fluid drip washing balance, the pH electrode of stirred-tank fermenter is inserted square opening on chromatography column simultaneously.
3. the external hose of substrate for enzymatic activity solution feed supplement bottle, demijohn and alkali bottle is accessed respectively the peristaltic pump of stirred-tank fermenter, wherein the flexible pipe outlet of soda acid feed supplement bottle is placed in the upper square opening of chromatography column, and substrate solution feed supplement bottle flexible pipe is connected with the lower square opening of chromatography column.
4. open the peristaltic pump be connected with substrate solution feed supplement bottle, promote substrate solution fast and enter chromatography column from lower square opening, immobilized enzyme is suspended, has fully contacted reaction with substrate solution.Start the mainboard Controlling System of stirred-tank fermenter, according to enzyme catalysis processing requirement, realize the control of temperature, pH and substrate feed supplement.
5. when substrate solution will be full of chromatography column material chamber, start external peristaltic pump, flow out a certain amount of reaction solution with constant rate of speed, ensure the unlikely spilling of substrate solution in chromatography column.
embodiment:
Embodiment below in conjunction with Production by Enzymes 6-amino-penicillanic acid and Semen Pisi sativi protein polypeptide is further described the present invention, and protection scope of the present invention is not by the restriction of embodiment.
Stirred-tank fermenter system major-minor self-contained, comprises fermentor tank main body, fermentor tank Controlling System, air compressor and air accumulator, thermostatic circulation bath, vapour generator, relies on silica gel hose to be interconnected between each appliance arrangement.
Embodiment 1: stirring-type biological fermentation tank is assembled into fluidized bed type enzyme reactor and produces 6-amino-penicillanic acid
Imported and exported with the chuck of the external chromatography column of silica gel hose respectively by the intake-outlet of the water bath circulation device of stirred-tank fermenter, effective length and width of chromatography column are respectively 40cm and 8cm.Add immobilized penicillin acylated enzyme (107U/g) by square opening on chromatography column in quiet heavy mode, and balance with the drip washing of pH8.0 damping fluid, the immobilized enzyme height after pre-equilibration is 1/4 of the actual use height of chromatography column.The pH electrode of stirred-tank fermenter is inserted square opening on chromatography column, the external hose of substrate for enzymatic activity solution feed supplement bottle, demijohn and alkali bottle is accessed respectively the peristaltic pump of stirred-tank fermenter simultaneously, wherein the flexible pipe outlet of soda acid feed supplement bottle is placed in the upper square opening of chromatography column, and substrate solution feed supplement bottle flexible pipe is connected with the lower square opening of chromatography column.Open the peristaltic pump that is connected with substrate solution feed supplement bottle, sample size with the speed of 60ml/min promote fast massfraction be 10% substrate solution enter chromatography column from lower square opening, by immobilized enzyme suspension, fully contacted reaction with substrate solution.When substrate solution will be full of chromatography column material chamber, start external peristaltic pump, flow out a certain amount of reaction solution with constant rate of speed, ensure the unlikely spilling of substrate solution in chromatography column.It is 40 DEG C that circulator bath controls temperature of reaction, and utilizing fermentor tank Controlling System to carry out automatic acid alkalimetric titration control reaction pH is 8.0, utilizes the yield of the product 6-amino-penicillanic acid of colorimetric method for determining chromatography column lower exit port to be 76%.
Embodiment 2: stirring-type biological fermentation tank is assembled into fluidized bed type enzyme reactor and produces Semen Pisi sativi protein polypeptide
Imported and exported with the chuck of the external chromatography column of silica gel hose respectively by the intake-outlet of the water bath circulation device of stirred-tank fermenter, effective use length and width of chromatography column are respectively 85cm and 10cm.Immobilization Validase TSP Concentrate II (352U/g), and balance with the drip washing of pH7.5 damping fluid, the immobilized enzyme height after pre-equilibration is 1/5 of the actual use height of chromatography column.The pH electrode of stirred-tank fermenter is inserted square opening on chromatography column, the external hose of substrate for enzymatic activity solution feed supplement bottle, demijohn and alkali bottle is accessed respectively the peristaltic pump of stirred-tank fermenter simultaneously, wherein the flexible pipe outlet of soda acid feed supplement bottle is placed in the upper square opening of chromatography column, and substrate solution feed supplement bottle flexible pipe is connected with the lower square opening of chromatography column.Open the peristaltic pump that is connected with substrate solution feed supplement bottle, sample size with the speed of 65ml/min promote fast massfraction be 6% substrate solution enter chromatography column from lower square opening, by immobilized enzyme suspension, fully contacted reaction with substrate solution.When substrate solution will be full of chromatography column material chamber, start external peristaltic pump, flow out a certain amount of reaction solution with constant rate of speed, ensure the unlikely spilling of substrate solution in chromatography column.It is 48 DEG C that circulator bath controls temperature of reaction, and utilizing fermentor tank Controlling System to carry out automatic acid alkalimetric titration control reaction pH is 7.5, and the yield utilizing ninhydrin colorimetry to measure the product Semen Pisi sativi protein polypeptide of chromatography column lower exit port is 24%.
Claims (1)
1. stirred-tank fermenter system assembles is the method for fluidized bed type enzyme reactor by one kind, it is characterized in that utilizing stirred-tank fermenter original system, the external double-jacket chromatography column with thermostatical circulating water can be passed in and out of fermentor tank silica gel hose, the bottom of chromatography column and 10cm place, upper end all have filters core for inclusive immobilization enzyme, and it specifically comprises the steps: that the fermentor tank main body of stirred-tank fermenter system, Controlling System, water bath circulation device, vapour generator, air compressor pump and air accumulator all possess; The intake-outlet of the water bath circulation device of stirred-tank fermenter is imported and exported with the chuck of the external chromatography column of silica gel hose respectively; Add immobilized enzyme by square opening on chromatography column in quiet heavy mode, and by damping fluid drip washing balance, the pH electrode of stirred-tank fermenter is inserted square opening on chromatography column simultaneously; The external hose of substrate for enzymatic activity solution feed supplement bottle, demijohn and alkali bottle is accessed respectively the peristaltic pump of stirred-tank fermenter, wherein the flexible pipe outlet of soda acid feed supplement bottle is placed in the upper square opening of chromatography column, and substrate solution feed supplement bottle flexible pipe is connected with the lower square opening of chromatography column; Open the peristaltic pump be connected with substrate solution feed supplement bottle, promote substrate solution fast and enter chromatography column from lower square opening, immobilized enzyme is suspended, has fully contacted reaction with substrate solution; When substrate solution will be full of chromatography column material chamber, start external peristaltic pump, flow out a certain amount of reaction solution with constant rate of speed, ensure the unlikely spilling of substrate solution in chromatography column.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201410420193.6A CN104178421B (en) | 2014-08-25 | 2014-08-25 | A kind of is the method for fluidized bed type enzyme reactor by stirred-tank fermenter system assembles |
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| CN201410420193.6A CN104178421B (en) | 2014-08-25 | 2014-08-25 | A kind of is the method for fluidized bed type enzyme reactor by stirred-tank fermenter system assembles |
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| CN104178421A CN104178421A (en) | 2014-12-03 |
| CN104178421B true CN104178421B (en) | 2016-01-13 |
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| CN106834117B (en) * | 2015-10-27 | 2018-11-23 | 苏州迪欧益生物科技有限公司 | For releasing the enzyme reactor and intelligent enzyme reaction work station of zooblast attaching |
| CN106635787A (en) * | 2016-11-07 | 2017-05-10 | 鲁东大学 | Method for assembling stir-type fermentation tank system into bubble column reactor |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US4430348A (en) * | 1980-01-29 | 1984-02-07 | Miller Brewing Company | Immobilized glucoamylase reactor for preparing a low calorie beer |
| CN1407103A (en) * | 2001-08-31 | 2003-04-02 | 中国科学院上海生物化学研究所 | Pearl chitin carrier preparation and process for using it for enzyme solidification |
| CN101182455B (en) * | 2007-11-29 | 2011-12-28 | 南京工业大学 | Device and process for producing propionic acid by coupling fiber bed reactor and extraction separation |
| CN101402914B (en) * | 2008-11-20 | 2011-08-31 | 南京工业大学 | Application of bagasse as immobilized material in immobilized fiber bed reactor for producing organic acid by fermentation |
| CN102146340A (en) * | 2010-12-30 | 2011-08-10 | 威海三友生物科技有限公司 | Fluidized bed reactor for stirring spherical immobilized enzyme |
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Effective date of registration: 20210926 Address after: 261400 No. 1158, Kaiming Road, Laizhou Development Zone, Yantai City, Shandong Province Patentee after: SHANDONG SEASIR BREWING FOOD Co.,Ltd. Address before: 264025 School of life sciences, Ludong University, 186 Hongqi Middle Road, Zhifu District, Yantai City, Shandong Province Patentee before: LUDONG University |