CN110747104A - Continuous in-situ separation device - Google Patents
Continuous in-situ separation device Download PDFInfo
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- CN110747104A CN110747104A CN201911173803.6A CN201911173803A CN110747104A CN 110747104 A CN110747104 A CN 110747104A CN 201911173803 A CN201911173803 A CN 201911173803A CN 110747104 A CN110747104 A CN 110747104A
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- HVCOBJNICQPDBP-UHFFFAOYSA-N 3-[3-[3,5-dihydroxy-6-methyl-4-(3,4,5-trihydroxy-6-methyloxan-2-yl)oxyoxan-2-yl]oxydecanoyloxy]decanoic acid;hydrate Chemical compound O.OC1C(OC(CC(=O)OC(CCCCCCC)CC(O)=O)CCCCCCC)OC(C)C(O)C1OC1C(O)C(O)C(O)C(C)O1 HVCOBJNICQPDBP-UHFFFAOYSA-N 0.000 description 1
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- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 description 1
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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
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/18—External loop; Means for reintroduction of fermented biomass or liquid percolate
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D21/00—Separation of suspended solid particles from liquids by sedimentation
- B01D21/24—Feed or discharge mechanisms for settling tanks
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D21/00—Separation of suspended solid particles from liquids by sedimentation
- B01D21/30—Control equipment
- B01D21/34—Controlling the feed distribution; Controlling the liquid level ; Control of process parameters
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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
- C12M39/00—Means for cleaning the apparatus or avoiding unwanted deposits of microorganisms
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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
- C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
- C12M47/10—Separation or concentration of fermentation products
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Abstract
The invention belongs to the technical field of fermentation, and particularly relates to a continuous in-situ separation device which comprises a shell and a shell inner cavity, wherein a partition plate is arranged at the lower part of the shell inner cavity and divides the shell inner cavity into an upper liquid separation cabin and a lower standing cabin; an outlet pipe and an inlet pipe are arranged on the shell body at the corresponding position of the liquid separation bin, the outlet pipe is positioned at the upper part of the inlet pipe, at least one layer of liquid separation baffle is arranged in the liquid separation bin between the outlet pipe and the inlet pipe, each layer of liquid separation baffle is inclined at an angle of 30-60 degrees relative to the horizontal plane, an overflow port and a reflux port are arranged on each layer of liquid separation baffle, and the reflux port of the liquid separation baffle positioned at the bottommost layer from top to bottom is communicated with the standing bin through a reflux; the diversion channel arranged on the partition plate is communicated with the liquid separation baffle plate at the bottommost layer and the liquid separation bin between the partition plates; the bottom of the shell is provided with a bottom valve which is communicated with the standing bin. The device of the invention ensures that the fermentation liquor is always in a circulating state, reduces the loss of thalli, reduces the risk of bacterial contamination and is easy for industrial production.
Description
Technical Field
The invention belongs to the technical field of fermentation, and particularly relates to a continuous in-situ separation device.
Background
The in-situ separation is also called as in-situ separation coupling technology, which is a fermentation technology that a product is continuously separated from a culture medium by using a separation and purification means in the fermentation or biological reaction process, so that the fermentation process and the separation process are synchronously performed, the product generated in the culture medium is separated in time, the inhibition effect of the product is further built, and the comprehensive yield and the productivity of the fermentation are integrally increased.
The in-situ separation technology comprises an adsorption method, a liquid-liquid extraction method, a gas stripping method, a pervaporation method, a membrane separation method and the like according to different separation means. The adsorption method is to adsorb the product in the fermentation medium by using an adsorption material such as macroporous resin, activated carbon and the like in the fermentation process so as to reduce the product inhibition effect in the fermentation process. For example, in situ separation of propionic acid coupled VB in published resins12Study of fermentation Process [ D]In the method, an in-situ separation mode of resin adsorption is adopted, so that the product inhibition of propionic acid on propionibacterium freudenreichii is greatly reduced, and the VB is improved12The yield of (2). The liquid-liquid extraction method adopts the method of adding an extraction phase and separating the fermentation product from the fermentation liquor in the fermentation process in an extraction mode, thereby reducing the product inhibition in the culture medium in the whole biological reaction process. For example, in the published literature of 2-phenylethyl alcohol synthesized by a biotransformation method in a water-polypropylene glycol two-phase system (proceedings of 2011,11(5):782 and 785), the 2-phenylethyl alcohol in the fermentation liquor is extracted by using polypropylene glycol two liquid phases, so that the yield of the 2-phenylethyl alcohol in the saccharomyces cerevisiae is improved. The membrane separation is a membrane separation method, for example, chinese patent application (CN103695475A) uses a vapor permeable membrane to collect and concentrate ethanol vapor volatilized from the upper part of the culture medium after vacuum pressure reduction by a membrane module method, so as to reduce the ethanol content in the fermentation medium and increase the production of bioethanol.
Sophorolipid (Sophorolipid) is a microbial secondary metabolite produced by candida with sugar, vegetable oil and the like as carbon sources through a fermentation process under certain conditions. Sophorolipid is a glycolipid biosurfactant. The surfactant has the general performances of solubilization, emulsification, wetting, foaming, dispersion, surface tension reduction and the like of the conventional surfactant, and also has the characteristics of no toxicity, 100% biodegradability, temperature resistance, high salt resistance, wide pH range adaptation, environmental friendliness and the like. The sophorolipid can be applied to the fields of petroleum, environmental protection, medicine, food, cosmetics, washing, home care, agriculture, feed and the like, and can partially or completely replace a chemically synthesized surfactant for use. The sophorose ester has the condition of multi-phase distribution in the fermentation process, including an oil phase, a lactone-type sophorose ester phase and a fermentation liquid phase, and when the multi-phase distribution is mixed, the sophorose ester has the condition of product inhibition, and simultaneously, the sophorose ester can weaken the dissolved oxygen mass transfer in a culture system. Therefore, the in-situ separation method is adopted, so that the yield and the production efficiency of the sophorose ester can be effectively increased.
An in-situ separation technique and a gravity separation device for fermentation production of sophorose ester are disclosed in the literature (Dolman, b.m., Kaisermann, c., Martin, p.j., & Winterburn, J.B. (2017). Integrated sophorolipid production and gravity separation. process biochemistry,54,162-171. doi:10.1016/j. procbio.2016.12.021), wherein sophorose ester is intermittently operated, that is, fermentation broth is pumped out of the gravity separation device and is allowed to stand for layering, and after the separation interface between the sophorose ester and the fermentation broth is clear, the fermentation broth is pumped back into the reactor through an upper or lower outlet of the device to continue fermentation while the sophorose ester product is also separated and collected. The publication (Liu, Z., Tian, X., Chen, Y., Lin, Y., Mohsin, A., & Chu, J. (2018) instant reagents production via a novel separation technology by Starmerella bombicola. Process biochemistry.doi:10.1016/J. procbio.2018.12.005) improves this one sophorose ester in-situ separation process on the basis of the disclosed in-situ separation scheme, which adds a pre-separation device before the gravity separation device.
In the known in-situ separation scheme for fermentation production of sophorose ester, the in-situ separation process is carried out intermittently, i.e. after a part of fermentation liquor is transferred into a separation device, standing and gravity separation are carried out, so that a lactone-type sophorose ester layer can be layered with the fermentation liquor, then the lactone-type sophorose ester is discharged from the separation device, then the fermentation liquor is discharged and returned to a fermentation tank, and the next step of separation can be carried out after the separation device is emptied. The prior technical scheme for in-situ separation of sophorose ester has the following problems:
(1) the intermittent batch operation type is not favorable for the continuous industrial production mode, and is difficult to be applied in the industrial production process.
(2) Intermittent batch operation increases a large amount of operation workload.
(3) Because the separation device communicated with the fermentation system needs to be emptied for many times, the probability of contamination is greatly increased.
(4) The standing gravity separation process requires a long standing and layering time.
(5) The thalli can also be precipitated in the standing gravity separation process, and the loss of the thalli amount can be caused in the step of emptying the separation device, so that the production efficiency is influenced.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a continuous in-situ separation device which can be applied to the fermentation production of sophorose ester in an in-situ separation mode and the fermentation process of similar products (such as rhamnose ester) so as to reduce the product inhibition in the fermentation process and improve the production efficiency.
The invention has the technical scheme that the continuous in-situ separation device comprises a shell (1) and a shell inner cavity wrapped by the shell (1), wherein a partition plate (6) is arranged at the lower part of the shell inner cavity, the shell inner cavity is divided into an upper liquid distribution bin (9) and a lower standing bin (8) by the partition plate (6), and a flow guide channel is arranged on the partition plate (6) and communicated with the liquid distribution bin (9) and the standing bin (8);
an outlet pipe (2) and an inlet pipe (3) are arranged on a shell (1) at a position corresponding to the liquid distribution bin (9), the outlet pipe (2) is positioned at the upper part of the inlet pipe (3), at least one layer of liquid distribution baffle (5) is arranged in the liquid distribution bin (9) between the outlet pipe (2) and the inlet pipe (3), the liquid distribution baffle (5) divides the liquid distribution bin (9) into more than two independent spaces, each layer of liquid distribution baffle (5) is parallel to each other, each layer of liquid distribution baffle (5) is inclined at an angle of 30-60 degrees relative to the horizontal plane, an overflow port (5.1) and a return port (5.2) are arranged on each layer of liquid distribution baffle (5), the overflow port (5.1) is higher than the return port (5.2), and the return port (5.2) of the liquid distribution baffle (5) positioned at the bottommost layer is communicated with a standing bin (8) through a return conduit (5.3; a diversion channel arranged on the partition plate (6) is communicated with a liquid separation bin (9) between the liquid separation baffle plate (5) at the bottommost layer and the partition plate (6);
the bottom of the shell (1) is provided with a bottom valve (4), and the bottom valve (4) is communicated with a standing bin (8).
In the in-situ separation and fermentation process of the sophorose ester, fermentation liquor in a fermentation tank is input from an inlet pipe (3) of an in-situ separation device, passes through an overflow port (5.1) of a multilayer liquid separation baffle plate (5) and a space between liquid separation baffle plates, and finally continuously flows back to the fermentation tank through an outlet pipe (2) arranged at the upper part of the separation device. The in-situ separation device and the fermentation tank form a closed continuous external circulation system. In the fermentation process, the circulating fermentation liquor generates coalescence in a narrow space between the liquid separating baffles, dispersed lactone type sophorose ester small oil drops gradually coalesce into large liquid drops, the specific gravity of the lactone type sophorose ester in a fermentation liquor system is higher than that of a culture medium, the coalesced lactone type sophorose ester can flow downwards along the obliquely arranged liquid separating baffles, the lactone type sophorose ester is enriched at the bottom of the slope of each layer of liquid separating baffles and flows back to the lower layer through a backflow port (5.2) arranged at the bottoms of the liquid separating baffles, the backflow port (5.2) of the bottommost layer of liquid separating baffles is directly connected with a backflow conduit (5.3), and finally the enriched lactone type sophorose ester flows back to a standing bin (8) at the bottom of the separating device through the backflow conduit (5.3). During the fermentation process, the liquid layer height of the lactone-type sophorose ester can be observed through a liquid level window (7) arranged at the bottom. And in the standing bin, after the lactone type sophorose ester liquid layer is highly filled, part of sophorose ester is discharged through a bottom valve, and the method is simple and convenient to operate. A small amount of lactone type sophorose ester liquid layer can be reserved in the operation process, and the lactone type sophorose ester liquid layer also plays a role in isolating fermentation liquor, so that the risk of bacterial contamination of a system in the process of draining and collecting sophorose ester is greatly reduced.
The partition board (6) divides the space inside the in-situ separation device into a liquid separation bin (9) in which the fermentation liquor continuously and circularly flows and a relatively static standing bin (8), and the lactone type sophorose ester which is collected and refluxed through coalescence can further stand and layer in the standing bin. The water phase of the fermentation liquor is distributed on the upper part of the lactone type sophorose ester layer, overflows back into the liquid separation bin through a communicating hole (6.1) arranged on the partition plate (6) or a flow guide port of a chimney type flow guide port (6.2), and returns into the continuous circulation of the whole fermentation liquor.
And a sight glass (7) is arranged on the shell corresponding to the standing bin (8), so that the condition in the standing bin (8) can be observed conveniently and timely.
The flow guide channel arranged on the partition board (6) is a plurality of communicating holes (6.1) and/or chimney-shaped flow guide openings (6.2). That is, the plurality of communication holes (6.1) may be provided, the chimney-type diversion opening (6.2) may be provided, the plurality of communication holes (6.1) and the chimney-type diversion opening (6.2) may be provided simultaneously, and the plurality of communication holes (6.1) or the plurality of communication holes (6.1) and the chimney-type diversion opening (6.2) may be provided simultaneously.
The distance between every two layers of separating baffles (5) is 5-20cm, preferably 8-15cm, and more preferably 10 cm. The number of layers of the separating baffle (5) is preferably 2-5, the embodiment is 3-4, and the number of layers can be 5, 6, 7, 8, 9, 10 or more according to the size of the actual separating device. Each layer of the separating baffle is preferably inclined at an angle of 45 degrees relative to the horizontal.
The overflow port (5.1) arranged on each layer of the liquid separating baffle (5) is positioned at the highest position of the liquid separating baffle (5), and the return port (5.2) is positioned at the lowest position of the liquid separating baffle (5).
An overflow drainage cover is fixed on the overflow port (5.1), the overflow drainage cover is positioned on one surface, close to the upper-end liquid separation baffle (5), of the liquid separation baffle (5), the overflow drainage cover can be a side opening or a top opening, the side opening is preferably selected, the side opening has a higher turbulence condition and a longer fluid flow path, and the occurrence of the sophorose ester oil phase coalescence process is facilitated.
An oil-water separation device is fixed at an opening on the side surface or the opening on the top end of the overflow drainage cover, so that the oil-water separation efficiency is improved. The oil-water separation device is a laminated grid net or a laminated corrugated plate, so that the syneresis and separation effect of sophorose is improved, the corrugated plate or the multi-layer grid net is filled in each layer of overflow port (2) to serve as a coalescer, so that the syneresis process of oil drops of the lactone sophorose ester oil phase is increased, particularly, the corrugated plate filler and the multi-layer grid net provide a tortuous channel for fluid to flow back and forth on the upper surface, and the syneresis process of the lactone sophorose ester liquid drops is accelerated when fermentation liquor passes through the overflow port.
The above-mentionedThe equivalent diameter of the grid in the grid mesh is 3-5 mm. The equivalent diameter means that if the meshes of the grid mesh are circular, the equivalent diameter should be the diameter of the circle. If the meshes of the grid net are square, the equivalent diameter is the side length; if the meshes of the grid net are in other special shapes, the annual diameter is the side length of a square with the same area corresponding to the area of the meshes, namely D is S1/2S is the mesh area and D is the equivalent diameter.
The more than two independent spaces divided by the liquid dividing baffle (5) into the liquid dividing bin (9) are respectively provided with laminated corrugated plates, so that stronger turbulent disturbance and more tortuous fluid flow paths are provided, and the separation efficiency of the in-situ separation device is greatly increased.
Be fixed with backward flow drainage cover on backward flow mouth (5.2), backward flow drainage cover is located the one side of keeping away from upper end liquid distribution baffle (5) of place liquid distribution baffle (5), and backward flow drainage cover can be the side opening also can be the bottom opening, preferred bottom opening.
The bottom valve (4) is communicated with the storage tank (10), the storage tank (10) is a sterilizable sealing tank body, and the sophorose ester separated in situ in the fermentation process is discharged into the storage tank (10) through the bottom valve (4), so that the risk of bacterial contamination is further reduced.
Compared with the technical scheme of the intermittent in-situ separation device in the prior art (Dolman, B.M., Kaisermann, C., Martin, P.J., & Winterburn, J.B. (2017) & Integrated Sophorrolipid production and visibility classification. Process Biochemistry,54,162-171. doi:10.1016/j. procbio.2016.12.021), the continuous in-situ separation device provided by the invention has the following remarkable technical effects:
(1) different from the existing in-situ separation scheme that the liquid filling, standing separation and emptying of the separation device need to be repeated for many times, the technical scheme provided by the invention does not need a complicated operation process. The whole fermentation liquor is always in a circulating state, only the separated lactone sophorolipid needs to be discharged through a bottom valve regularly, and the method is simple and convenient and is easy for industrial production.
(2) Compared with the prior in-situ separation scheme, the technical scheme provided by the invention has the advantages that the fermentation liquor is always in a dynamic circulation process and does not have a standing process, so the thallus loss in the in-situ separation process caused by cell precipitation is greatly less than that in the prior technical scheme.
Compared with the prior art, the technical scheme provided by the invention does not need to empty the separation device for multiple times, and has lower risk of bacterial contamination.
Drawings
FIG. 1 is a schematic sectional view of an in-situ separation apparatus according to example 1.
FIG. 2 is a schematic diagram of the structure of a closed circulation system among a fermentation tank, an in-situ separation device and a storage tank.
FIG. 3 shows the distribution and layering of vegetable oil, fermentation broth, and lactone-type sophorose ester in the fermentation process of example 2 (from left to right, the fermentation broth samples are sequentially layered after fermentation for 1 to 10 days, wherein the lactone-type sophorose ester layer always settles at the bottom of the liquid separation layer).
FIG. 4 is a schematic view of an in-situ separation apparatus in which a plurality of communication holes are formed in a partition plate in the in-situ separation apparatus according to example 3.
FIG. 5 is a schematic view showing the type of the overflow holes filled in the side-open overflow port and the structure thereof in example 4 (from left to right, the side-open overflow holes, the overflow holes filled in the multi-layer grid mesh, and the overflow holes filled in the corrugated plate, respectively)
Detailed Description
Example 1 continuous in-situ separation, fermentation and separation device for sophorose ester and use method thereof
The in-situ separation device adopted in the embodiment is shown in the attached figure 1 and comprises a shell 1, an outlet pipe 2, an inlet pipe 3, a bottom valve 4, a liquid separation baffle plate 5, a partition plate 6 and a liquid level window 7; 1 bottom of casing is provided with bottom valve 4, baffle 6 separates the casing inner chamber into two independent storehouse bodies, for the branch liquid storehouse 9 on storehouse 8 and the upper portion of stewing of lower part, it is equipped with liquid level window 7 on the casing 1 at storehouse 8 place to stew, 6 upper portions of baffle are equipped with import pipe 3, be provided with outlet pipe 2 on the upper portion of casing 1 simultaneously, be provided with three-layer branch liquid baffle 5 between outlet pipe 2 and the import pipe 3, every layer divides liquid baffle 5 and level to be 45 degrees contained angle slope arrangement in the cavity between outlet pipe 2 and import pipe 3, every layer divides liquid baffle 5 to be parallel to each other, and the interval between every layer divides liquid baffle 5 is 10cm, be equipped with overflow mouth 5.1 and backward flow mouth 5.2 on every layer divides liquid baffle 5 simultaneously, set up in 5 backward flow mouth 5.2 of backward flow on the liquid baffle of bottommost directly the drainage to the storehouse 8 of stewing through. The opening mode of the overflow port 5.1 is a side opening (as shown in figures 1 and 4), and the opening mode of the return port 5.2 is a straight opening (as shown in figures 1 and 4). The connection mode between the in-situ separation device and the fermentation tank and the storage tank 10 is shown in the attached figure 2, in the in-situ separation and fermentation process of the sophorose ester, fermentation liquor is input from an inlet pipe 3 of the in-situ separation device and continuously flows back to the fermentation tank through an outlet pipe 2 arranged at the upper part of the separation device through a space between an overflow port 5.1 of a multi-layer liquid separation baffle plate and the liquid separation baffle plate in the fermentation process. The in-situ separation device and the fermentation tank form a closed continuous external circulation system. In the fermentation process, the circulating fermentation liquor generates coalescence in a narrow space between the liquid separating baffles, dispersed lactone type sophorose ester small oil drops gradually coalesce into large liquid drops, the specific gravity of the lactone type sophorose ester in a fermentation liquor system is higher than that of a culture medium, the coalesced lactone sophorose ester can flow downwards along the obliquely arranged liquid separating baffles, is enriched at the bottom of the slope of each layer of liquid separating baffles and flows back to the lower layer through a backflow port 5.2 arranged at the bottoms of the liquid separating baffles, the backflow port 5.2 of the liquid separating baffle at the bottommost layer is directly connected with a backflow conduit 5.3, and finally the enriched lactone type sophorose ester flows back to the standing bin 8 at the bottom of the separating device through the backflow conduit. During the fermentation process, the liquid layer height of the lactone-type sophorose ester can be observed through a liquid level window 7 arranged at the bottom. And after the lactone type sophorose ester liquid layer is highly filled in the standing bin, discharging sophorose ester through a bottom valve. The liquid conveying process between the fermentation tank and the in-situ separation device adopts a peristaltic pump as power.
Example 2 continuous in situ separation fermentation of sophorose esters
Since the lactone-type sophorose ester is separated from the bottom valve of the continuous in-situ separation apparatus when the fermentation is performed by the continuous in-situ separation method provided in example 1, the specific gravity of the fermentation medium and the fermentation liquid is controlled to be lower than that of the lactone-type sophorose ester during the whole fermentation period. The present embodiment was carried out by the following culture method: candida ATCC22214 was selected as a production strain, which was inoculated into a 1L shake flask (liquid loading 200mL medium with 2% w/v glucose, 2% w/v peptone, 1% w/v yeast powder, pH5.0) and shake-cultured at 30 ℃ for 24h as seed liquid. The liquid loading amount of a 5L fermentation tank serving as a culture tank body is 3L (fermentation culture medium: glucose 6% w/v; rapeseed oil 1% w/v; yeast powder 0.3% w/v; disodium hydrogen phosphate dodecahydrate 0.1% w/v; potassium dihydrogen phosphate 0.1% w/v; magnesium sulfate heptahydrate 0.05% w/v; initial pH5.0), the seed liquid is inoculated into the tank according to the inoculation amount of 1%, the dissolved oxygen is controlled to be 50%, the dissolved oxygen is linked with the rotating speed (the upper limit and the lower limit of the rotating speed are 300-800rpm), the fermentation temperature is 25 ℃, and the rapeseed oil is continuously supplemented according to the speed of 3.75g/h during the fermentation period; the water replenishing rate of the material replenishing pipe is as follows: 3.35mL/min while continuously adding a glucose solution (concentration: 800g/L) at a rate of 10 g/L/day. The experimental group adopted in-situ separation fermentation with the in-situ separation apparatus shown in the example 1, and the peristaltic pump set the peristaltic delivery rate of 100 mL/h. Meanwhile, a fermentation tank fermentation device which does not adopt an in-situ separation device is provided with the same fermentation conditions for fermentation as a control. As a result, according to the culture conditions of the present embodiment, the specific gravity of the sophorose ester liquid layer is higher than that of the fermentation liquid (or fermentation medium) during the whole fermentation period (see FIG. 3), so that the lactone-type sophorose ester liquid can settle at the bottom of the standing zone of the in-situ separation apparatus, and can be smoothly separated from the bottom valve of the in-situ separation apparatus. The total yield of lactone sophorose ester in the experimental group adopting the in-situ separation device reaches 82g/L, and the total yield of lactone sophorose ester which can be collected in the culture medium without adopting the in-situ separation device is only 41 g/L.
Example 3 improvement of the in-situ separation apparatus in example 1
In the continuous in-situ separation apparatus of example 1, the partition 6 divides the in-situ separation apparatus into a standing bin 9 and a liquid separation bin 8. Wherein, the liquid in the liquid separating bin is always in a motion circulation state, while the liquid in the standing bin is relatively static, and can flow back to the liquid separating bin only through a chimney type diversion port 6.2 arranged on the partition plate 6. Because the liquid in the liquid separation bin is relatively static, partial thalli can be settled and irradiated into partial thalli to be lost. As shown in fig. 4, the chimney-type diversion port 6.2 in the embodiment 1 is changed to be arranged on a plurality of connection holes 6.1 on the partition plate. The improvement increases the fluid communication of the standing bin 9 and the liquid separating bin 8 at the boundary, and can reduce the thallus precipitation by about 30-40%.
Example 4 improvement of the in-situ separation apparatus in example 1
The coalescence efficiency of small oil drops (i.e. lactone type sophorose ester liquid drops) in the circulation process of the fermentation liquor determines the in-situ separation device of the whole device. The use of stacked corrugated sheets or multi-layer grid mesh is effective in providing greater turbulence and more tortuous fluid flow paths to greatly increase the separation efficiency of the in situ separation device. As shown in fig. 5, the present embodiment provides an improvement manner of the in-situ separation device, that is, stacked corrugated plates or multiple layers of grid nets are filled at the position of the overflow port of the in-situ separation device to serve as an oil-water separator, so as to improve the separation efficiency of the lactone-type sophorose ester.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. A continuous in-situ separation device is characterized by comprising a shell (1) and a shell inner cavity wrapped by the shell (1), wherein a partition plate (6) is arranged at the lower part of the shell inner cavity, the shell inner cavity is divided into an upper liquid distribution bin (9) and a lower standing bin (8) by the partition plate (6), a flow guide channel is arranged on the partition plate (6), and the flow guide channel is communicated with the liquid distribution bin (9) and the standing bin (8);
an outlet pipe (2) and an inlet pipe (3) are arranged on a shell (1) at a position corresponding to the liquid distribution bin (9), the outlet pipe (2) is positioned at the upper part of the inlet pipe (3), at least one layer of liquid distribution baffle (5) is arranged in the liquid distribution bin (9) between the outlet pipe (2) and the inlet pipe (3), the liquid distribution baffle (5) divides the liquid distribution bin (9) into more than two independent spaces, each layer of liquid distribution baffle (5) is parallel to each other, each layer of liquid distribution baffle (5) is inclined at an angle of 30-60 degrees relative to the horizontal plane, an overflow port (5.1) and a return port (5.2) are arranged on each layer of liquid distribution baffle (5), the overflow port (5.1) is higher than the return port (5.2), and the return port (5.2) of the liquid distribution baffle (5) positioned at the bottommost layer is communicated with a standing bin (8) through a return conduit (5.3; a diversion channel arranged on the partition plate (6) is communicated with a liquid separation bin (9) between the liquid separation baffle plate (5) at the bottommost layer and the partition plate (6);
the bottom of the shell (1) is provided with a bottom valve (4), and the bottom valve (4) is communicated with a standing bin (8).
2. The continuous in-situ separation device according to claim 1, characterized in that a viewing mirror (7) is arranged on the housing corresponding to the standing bin (8) to facilitate timely observation of the conditions in the standing bin (8).
3. Continuous in-situ separation device according to claim 1, characterized in that the flow guiding channels provided on the partition (6) are a plurality of communication holes (6.1) and/or chimney-type flow guiding openings (6.2).
4. The continuous in-situ separation device according to claim 1, wherein the distance between each layer of separating baffles (5) is 5-20 cm.
5. The continuous in-situ separation device according to claim 1, wherein the number of layers of the separating baffle (5) is 2-5.
6. The continuous in-situ separation device according to claim 1, characterized in that the overflow port (5.1) arranged on each layer of the separating baffle (5) is positioned at the highest position of the separating baffle (5), and the return port (5.2) is positioned at the lowest position of the separating baffle (5).
7. The continuous in-situ separation device according to claim 1, characterized in that an overflow drainage cover is fixed on the overflow port (5.1), the overflow drainage cover is positioned on one surface of the liquid separation baffle (5) close to the upper end liquid separation baffle (5), and the overflow drainage cover is open at the side surface or open at the bottom end;
a backflow drainage cover is fixed on the backflow port (5.2), the backflow drainage cover is located on one side, away from the upper end, of the liquid distribution baffle (5), and the backflow drainage cover is open on the side face or the bottom end.
8. The continuous in-situ separation device of claim 7, wherein an oil-water separation device is fixed at the opening of the side surface or the opening of the top end of the overflow drainage cover, and the oil-water separation device is a stacked grid net or a stacked corrugated plate.
9. The continuous in-situ separation device according to claim 1, wherein the separation baffle (5) divides the separation bin (9) into more than two independent spaces, and laminated corrugated plates are arranged in each space.
10. Continuous in-situ separation apparatus according to claim 1, characterized in that the bottom valve (4) communicates with a storage tank (10).
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111807592A (en) * | 2020-07-17 | 2020-10-23 | 河北兰升生物科技有限公司 | Continuous liquid separation device and organic solvent separation method using same |
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