WO2022154690A1 - Bioréacteur à tourbillons de gaz à plateformes multiples de laboratoire - Google Patents

Bioréacteur à tourbillons de gaz à plateformes multiples de laboratoire Download PDF

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Publication number
WO2022154690A1
WO2022154690A1 PCT/RU2021/000512 RU2021000512W WO2022154690A1 WO 2022154690 A1 WO2022154690 A1 WO 2022154690A1 RU 2021000512 W RU2021000512 W RU 2021000512W WO 2022154690 A1 WO2022154690 A1 WO 2022154690A1
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Prior art keywords
annular
annular floating
partition
container
medium
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PCT/RU2021/000512
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English (en)
Russian (ru)
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Андрей Петрович РЕПКОВ
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Общество С Ограниченной Ответственностью "Центр Вихревых Технологий"
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Publication of WO2022154690A1 publication Critical patent/WO2022154690A1/fr

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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
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/02—Apparatus for enzymology or microbiology with agitation means; with heat exchange means
    • 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
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/04—Apparatus for enzymology or microbiology with gas introduction means
    • 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
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/34—Measuring or testing with condition measuring or sensing means, e.g. colony counters
    • 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
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/36—Apparatus for enzymology or microbiology including condition or time responsive control, e.g. automatically controlled fermentors

Definitions

  • the invention relates to devices for cultivating lower phototropes (microalgae), as well as for carrying out biochemical processes using liquid media of various viscosities, in particular when cultivating tissue cells and microorganisms in nutrient media with increased or varying viscosity.
  • the invention can be used in the educational process at the biological faculties of various educational institutions (from schools to universities), as well as an experimental photo- and bioreactor (fermenter) in the organization of industrial production.
  • Known bioreactor containing a cylindrical container with a lid and nozzles for inlet and outlet of gas and a device for aeration and mixing of the medium.
  • the device for aeration and mixing contains a horizontal impeller mounted on a vertical drive shaft, placed in the upper part of the container directly under the lid, and an annular plate located under it with a central hole for passing gas, attached along the periphery to the wall of the container with the formation of an annular cavity around the wheel for gas inlet and outlet.
  • Slotted holes for gas passage are made in the annular partition, spaced evenly along the circumference at an inclination to the horizontal plane (WO 92105245, C12N 5/00, 02.04.1992).
  • this bioreactor is not universal in relation to laboratory biotechnological research and does not provide efficient mixing of a viscous culture medium during biochemical processes.
  • the presence of a guide apparatus around the activator leads to a loss of power of the gas vortex and, at the same time, to an increase in rarefaction in the axial zone of the bioreactor, as a result of which, at high speeds of gas movement above the surface of the latter, droplets with cultured cells are captured and thrown onto the container wall, which leads to injury and death, for example, of plant or animal cells.
  • "Drop separation" can be an effective mode only when cultivating, for example, yeast cultures.
  • a bioreactor including a cylindrical container with a lid, a device for mixing the medium located in it, consisting of a paddle wheel horizontally mounted on a vertical shaft in the upper part of the container, and an annular partition installed in the container with the possibility of rotation, forming a gap with the wall of the container and provided with floats, and a mechanism for adjusting the position of the annular partition in height relative to the mixed medium.
  • This mechanism may contain racks attached to the lid of the container and the annular partition using clamps and allowing you to change the position of the partition along the height of the container (RU 2099413 C 1 , S12M 1/04, 12/20/1997).
  • the disadvantage of the bioreactor is that when the fixed annular partition is attached, it creates a great resistance to the movement of the viscous culture medium, which worsens the process of its mixing.
  • the use of a floating annular baffle leads to the fact that it is ejected to the surface of the liquid and periodically fluctuates with its displacement to the vessel wall, which worsens the mixing process.
  • the design of the bioreactor is not universal in relation to laboratory biotechnological research
  • Known bioreactor (RU, 2299903, S12M 1/04, publ. 05/27/2007), including a cylindrical container with a lid, located in it a device for mixing the medium, consisting of a paddle wheel, horizontally mounted on a vertical shaft in the upper part of the container, and an annular partition installed in a container with the possibility of rotation, forming a gap with the wall of the container and equipped with floats, and a mechanism for adjusting the position of the annular partition in height relative to the surface of the mixed medium.
  • This mechanism contains a rod vertically mounted along the axis of the container and a sleeve placed on it, connected by means of racks with an annular partition for its rotation on the rod.
  • the mechanism for adjusting the position of the annular baffle can be provided with a lock of its position relative to the rod, containing an additional bushing located between the rod and the bushing of the annular baffle, the stroke limiters of the latter, and at least one clamping element.
  • the rod is installed with the possibility of its axial movement.
  • this reactor operates in liquid media, the viscosity of which does not exceed the viscosity of water by more than 2 times.
  • culturing plant and animal cells prone to agglomeration especially in a viscous medium or a medium with varying viscosity, there is a problem of settling of cells or cell clusters on the upper surface of the annular partition.
  • cell agglomerates fixed on the surface of the washer, the lower layers of cells die and are poisoned by the products of environmental lysis, which negatively affects the results of cultivation.
  • the design of this bioreactor is not universal in relation to laboratory biotechnological research.
  • a plant for cultivating lower phototrophs including a photobioreactor, a lighting system and an installation platform (RF Patent for utility model No. 150345, IPC C12N1/12, publ. 10.02.2015).
  • the installation is supplemented with a thermal stabilization system in the form of a water circuit rigidly connected to the photobioreactor and a system for ensuring a continuous cultivation regime containing containers for a nutrient medium, a programmable time switch and an electromagnetic valve, while the installation site consists of an upper and lower shelves, a lighting system located on the bottom shelf , includes horizontal light gratings located behind the photobioreactor.
  • the photobioreactor of a plane-parallel type with a volume of 3 liters, made with the possibility of moving relative to horizontal light gratings, has mirrored side walls and glass front walls, as well as a bottom located under angle 25°.
  • the design of the specified photobioreactor does not provide the possibility of scaling the cultivation technology to volumes of more than 3-5 liters, and is also difficult to maintain between cultivation processes (disassembly of the bioreactor, washing its internal volume, sterilization and assembly), and also does not provide the possibility of effective cultivation of plant and animals cell cultures.
  • the design of this bioreactor is not universal in relation to laboratory biotechnological research.
  • a known bioreactor for cultivating photosynthetic microorganisms containing a container with a lid and a device for mixing and aerating microorganisms, including nozzles, respectively, for supplying aerating gas and removing a gaseous medium, placed in the lid of the container, and configured to create a swirling flow of aerating gas on the surface of the suspension with a field speed of the potential vortex at the periphery of the tank and axial counterflow in the axial zone and the pressure drop between the periphery and the center of the vortex and several annular partitions with hollow floats open from below, installed in the tank coaxially with them on the vertical axis at a distance from each other with the possibility of rotation and reciprocating moving along it with the formation of a gap between the vessel wall and the annular partitions (RF patent No.
  • the lid, reactor vessel and annular partitions are made of optically transparent materials.
  • the bioreactor is equipped with sources of artificial lighting installed respectively in the cavities of the floats of the annular partitions.
  • the container lid has a rigid structure, and the container is made in the form of a disposable or reusable removable shell and is provided with means for attaching it to the lid and bottom of the reactor vessel, respectively.
  • the shell of the container is made of soft optically transparent elasto-elastic polymeric material.
  • 27.08.1999 containing a cylindrical container with a lid and nozzles, respectively, for supplying aerating gas and removing a gaseous medium and a device for aeration and mixing of the medium, including a horizontal impeller mounted on a vertical hollow shaft and placed in the upper part of the tank directly under the lid, an annular partition installed in the cylindrical container coaxially with the impeller with the formation of a gap between the cylindrical wall of the container and the annular partition, and a mechanism for stabilizing the position of the annular partition relative to the surface of the cell suspension, made in the form of guide elements and floats.
  • the above device is not universal and does not allow the cultivation of lower phototropes (microalgae).
  • the closest analogue is a bioreactor, including a cylindrical container with a lid, a device for mixing the medium located in it, consisting of a paddle wheel horizontally mounted on a vertical shaft in the upper part of the container, and a horizontal annular partition installed in the container with a gap relative to its cylindrical walls, a rod vertically mounted along the axis of the tank, on which a horizontal annular partition is placed with the possibility of rotation and a mechanism for fixing the horizontal annular partition on the rod (RF patent No. 2538170, IPC C12N1 / 12, publ.
  • the bioreactor is equipped with a pipe or a telescopic pipe docked with the axial hole of the horizontal annular baffle, attached from below to the latter and located around the rod, and in the horizontal annular baffle there are radial channels located from the axial hole to the edge of the annular partition with an inclination towards the bottom towards the bottom of the tank.
  • the horizontal annular partition is made floating, for example, from polypropylene.
  • the inner diameter of the pipe or telescopic pipe corresponds to the diameter of the axial hole of the horizontal annular baffle.
  • the area of the axial opening of the horizontal annular partition is equal to the total cross-sectional area at the inlet to the radial channels and the total cross-sectional area at the outlet of these radial channels.
  • the technical result is the creation of a more universal laboratory multiplatform bioreactor, which makes it possible to cultivate microorganisms, plant and animal cell cultures and microalgae (lower phototropes) in laboratory conditions, depending on the research tasks.
  • the bioreactor for suspension cultivation of tissue cells or microorganisms, containing a cylindrical container with a lid and nozzles, respectively, for supplying aerating gas and removing a gaseous medium and a device for aeration and mixing of the medium, including a horizontal impeller, reinforced on a vertical shaft and located in the upper part of the container directly under the lid, an annular floating partition installed in a cylindrical container with the possibility of rotation coaxially with the impeller with the formation of a gap between the cylindrical wall of the container and the annular floating partition, and a mechanism for stabilizing the position of the annular floating partition relative to the surface of the cell suspension, made in the form of floats, made on top of the body of the said annular partition with a gap relative to each other and forming radial channels between themselves, located from the axial hole to the edge of the said annular partition, according to the invention, the bioreactor is equipped with a removable optically transparent hollow
  • the light source is placed in a transparent hollow cylindrical cup with a gap and contains an LED strip fixed on the outer surface of a metal pipe and installed with an annular a gap in the glass pipe and a unit for forced cooling of the LED strip, made in the form of an axial fan, the outlet channel of which is connected to a metal pipe with an LED strip to form a circulating air flow around the LED strip.
  • the annular floating partition is made of one or two tiers, and the radial channels in each tier are located tangentially and directed counterclockwise.
  • the annular floating element may contain through holes for the flow of liquid nutrient medium, made at the same distance in the body of the annular floating element from the side of its cylindrical surface.
  • FIG. Figure 1 shows a diagram of a laboratory multiplatform gas-vortex bioreactor with a complete set of equipment (platform No. 1) designed for cultivating microalgae (lower phototropes).
  • FIG. Figure 2 shows a diagram of a laboratory multiplatform gas-vortex bioreactor without an annular floating element with a stirrer and needle elements for surface bubbling (platform No. 2).
  • Such a bioreactor is intended for the cultivation of bacteria and cell cultures on a laboratory scale that do not require increased aeration.
  • FIG. Figure 1 shows a diagram of a laboratory multiplatform gas-vortex bioreactor with a complete set of equipment (platform No. 1) designed for cultivating microalgae (lower phototropes).
  • FIG. Figure 2 shows a diagram of a laboratory multiplatform gas-vortex bioreactor without an annular floating element with a stirrer and needle elements for surface bubbling (platform No. 2).
  • Such a bioreactor is intended for the cultivation of bacteria and cell cultures on a laboratory scale
  • FIG. 3 shows a diagram of a laboratory multiplatform gas-vortex bioreactor without an internal transparent glass, an annular floating element with a stirrer, and needle elements for surface bubbling (platform No. 3).
  • This bioreactor is intended for laboratory and industrial cultivation of bacteria and cell cultures that do not require increased aeration.
  • FIG. 4 is given top view A of figure 5.
  • FIG. 5 shows a suspension mixing and aeration unit, including an annular floating baffle, an annular floating element with a stirrer and needle elements for surface bubbling.
  • FIG. 6 shows a side view B of figure 5.
  • FIG. 7 shows an annular floating element with a paddle agitator and needle elements for surface sparging.
  • FIG. 8 shows a diagram of the claimed laboratory multi-platform gas-vortex bioreactor (without an annular floating baffle 7) with an annular floating element with removable blades and needle elements (platform No. 4).
  • FIG. 9 shows a diagram of the claimed laboratory multiplatform gas-vortex bioreactor (without an annular floating baffle) with an annular floating element8 with removable blades (without needle elements for surface bubbling).
  • FIG. 10 shows a diagram of the claimed laboratory multi-platform gas-vortex bioreactor (without an annular floating baffle) with an annular floating element and needle elements (aerators) for surface bubbling (without removable blades).
  • the laboratory multiplatform gas-vortex bioreactor for suspension cultivation of tissue cells, microorganisms or microalgae contains an optically transparent cylindrical container 1 with a lid 2 and nozzles 3, 4, respectively, for supplying aerating gas and removing a gaseous medium, and a device for aeration and mixing of the medium, including a horizontal paddle wheel 5, mounted on a vertical shaft 6 and placed in the upper part of the container 1 directly under the cover 2, an annular floating partition 7 with a through axial hole 8, installed in the cylindrical container 1 with the possibility of rotation coaxially with the paddle wheel 5 with the formation of a gap between the cylindrical wall of the container 1 and an annular floating partition 7.
  • the shaft 6 is connected to the wheel rotation drive 5 located on the cover 2 of the container 1.
  • the mechanism for stabilizing the position of the annular floating partition 7 relative to the surface of the suspension of microorganism cells is made in the form of floats 9 located on top (single-tier partition 7, Fig. 1 ) or above and in the body of said annular partition 7 (two-tier partition 7, Fig. 5) with a gap relative to each other and forming radial channels 10 between them, located from the axial hole 8 to the edge of the said annular partition 7.
  • the bioreactor is equipped with a removable optically transparent hollow cylindrical glass 11, made with the possibility of placing in it a source 12 of illumination and / or a block 13 of video recording, measurement and control of the technological process of cultivation, and installed under the annular floating partition 7.
  • the lower open end of the glass 11 is hermetically attached to the bottom 14 of the container 1 and forms between cylindrical wall container 1 and the cylindrical wall of the glass 11 an annular zone 15 suspension cultivation of biological objects.
  • the mechanism for stabilizing the position of the annular floating partition 7 relative to the surface of the cell suspension is provided with an axis 16 installed coaxially with the horizontal paddle wheel 5 with the possibility of its attachment to the end wall 17 of the cylindrical cup 11.
  • the device for aeration and mixing of the medium (Fig. 1, 4-6) is equipped with an annular floating element 18 located in the gap between the cylindrical wall of the container 1 and the annular floating partition 7 with the possibility of rotation and containing (Fig. 1, 8-10):
  • the light source 12 is placed in a transparent hollow cylindrical cup 21 with a gap and contains an LED strip 22 fixed on the outer surface of a metal pipe 23 and a forced cooling unit for the LED strip 22, made in the form of an axial fan 24, the output channel of which is connected to the metal tube 23 of the LED strip 22 to form around the latter a circulating air flow (FIG. 8).
  • the annular floating partition 7 is made of one or two tiers, and the radial channels 10 in each tier are located tangentially and directed counterclockwise (Fig. 4-6). Said floating baffle 7 can be equipped with blades "B" for its additional unwinding by a vortex flow. Annular floating partition 7 ensures the functioning of the bioreactor in the optimal mode when filling its volume from 10 to 90%.
  • Single-tiered floating partition 7 can operate at a speed of rotation of the impeller 5 up to 2000 rpm. When the speed of rotation of the impeller 5 is over 2000 rpm, the central upward flow of the culture medium is able to push the single-tier floating partition 7 onto the surface of the specified medium. In this case, the flow of the upward flow of the culture is blocked.
  • a single-tier floating partition 7 returns to its original depth and the process of its ascent in this mode is repeated, which disrupts the cultivation process at high speeds of the impeller 5.
  • the two-tier floating partition 7 eliminates these disadvantages and allows the cultivation process to be carried out both at low speeds of the impeller 5, and at speeds over 2000-2500 rpm.
  • the annular floating element 18 (Fig. 7) may contain through holes 25 for the flow of liquid nutrient medium, made at the same distance in the body of the annular floating element 18 from the side of its cylindrical surface.
  • the annular floating element 18 with needle elements 20 for surface bubbling can be made of polypropylene (density 9.6 g/cm 3 ) with pointed conical protrusions on its upper plane 3-5 mm high. With the help of additional weights and floats, the floating element 18 is balanced so that, when immersed in the culture medium, the points of the cones are at the level of the medium surface. The floating element 18 in the process of cultivation is always in the zone of formation of the downward flow of the culture medium.
  • the gas vortex deforms the surface of the medium above the needle-shaped elements 20 in the form of conical protrusions behind each of which a plume of air bubbles is formed, which, before surfacing, have time to plunge into the medium to a depth of 5-10 mm, as a result of which it is saturated with gas (for example, oxygen or carbon dioxide).
  • gas for example, oxygen or carbon dioxide.
  • the gas-saturated medium descends in a spiral to the bottom of the bioreactor, providing aeration throughout the volume of the medium.
  • Suspension cultivation using the inventive laboratory multiplatform gas-vortex bioreactor is carried out as follows.
  • platform No. 1, 2, 3, 4, 5 or 6 of the bioreactor with different equipment is selected.
  • Example 1 For the cultivation of microalgae (lower phototropes) use platform No. 1 of the bioreactor (Fig. 1). Equipment is installed in container 1 under sterile conditions as shown in Fig. 1 and fill with a nutrient medium through a pipe (not shown in the drawing) so that above the surface of the medium in the upper part of the container 1 under the lid 2 there is a cavity for the movement of aerating gas, the annular partition 7 with floats 9 is located in the nutrient medium near its surface.
  • a seed dose of photosynthetic cells of microorganisms, such as chlorella is introduced (in the amount of 0.1 g per 1 liter of nutrient medium.), And the impeller 5 rotation drive is turned on, which supplies air enriched, for example, with carbon dioxide (for chlorella) through the pipe 3 tangentially into the container 1, and through the branch pipe 4 the exhaust gas.
  • the system for enriching the air with carbon dioxide is not shown in the drawing.
  • the required number of revolutions of the impeller 5 of the fan is set. Above the annular surface of the cell suspension, a vacuum is created in the paraxial zone of the container 1 and an increased pressure on the periphery of this container 1.
  • the speed of movement of the gas vortex (3-6 m/s) does not cause separation of drops of culture fluid from its surface, which reduces cell injury.
  • This provides optimal conditions for the cultivation of photosynthetic microorganisms.
  • the source 12 provides sufficient uniform illumination of the annular zone 15 of the chlorella suspension, the thickness of which is 30-50 mm.
  • the process is carried out continuously until the increase in the biomass of photosynthetic microorganisms, such as chlorella algae, up to 15-20 g per 1 liter of suspension, after which part of the suspension (crop) is drained, fresh nutrient medium is added and the cultivation process continues.
  • Example 2 To prepare a multiplatform gas-vortex bioreactor in the form of platform No. 2, an annular floating element 18 with a stirrer 19 and needle elements 20 for surface bubbling, as well as a light source 12, are removed from vessel 1.
  • Platform No. 2 allows cultivating bacteria and cell cultures in laboratory conditions , observing and recording the specified process using the video recording unit 13, measuring and controlling the technological process of cultivation (Fig. 2), which is very important when conducting scientific research in the laboratory.
  • the cultivation process is carried out as follows. Equipment is installed in container 1 under sterile conditions as shown in Fig.
  • a vacuum is created in the paraxial zone of the container 1 and an increased pressure on the periphery of this container 1.
  • a swirling gas flow is created above the annular surface of the cell suspension, which forms the culture liquid in the annular zone 15 turbulent rotational movement with intense mixing between the outer side of the cylindrical surface of the glass 11 and the inner surface of the vessel 1.
  • the gas aerates the cells on the surface of the liquid.
  • the speed of movement of the gas vortex (3-6 m/s) does not cause separation of drops of culture fluid from its surface, which reduces cell injury. This provides optimal conditions for the cultivation of plant or animal cells.
  • the process is carried out continuously until the growth of cell biomass, after which part of the suspension (harvest) is drained, fresh nutrient medium is added and the cultivation process is continued.
  • Example 3 Cultivation of bacteria, animal or plant cells on an industrial scale using platform #3 (FIG. 3).
  • an annular floating element 18 with a stirrer 19 and needle elements 20 is removed from the tank 1, and the glass is also dismantled AND .
  • a cylindrical container 1 with an annular floating partition 7 installed in it with blades "B" (Fig. 3) is filled under sterile conditions with a nutrient medium so that above the surface of the medium in the upper part of the container 1 remains a cavity for the movement of aerating gas, and the annular partition 7 was located in the nutrient medium with protruding blades "B" above the liquid medium.
  • the required temperature regime is set, the seed dose of cells is also introduced, and the rotation drive of the impeller 5 is turned on.
  • a vacuum is created in the axial zone of the container 1 and an increased pressure on the periphery of this container.
  • a swirling gas flow is created, which forms a turbulent rotational motion in the culture liquid with intense mixing along the axis of the container.
  • the annular floating partition 7 rotates in the same direction and with the same angular velocity as the culture liquid and is held near the surface of the liquid with blades "B" located above the surface of the liquid due to its own buoyancy.
  • the annular partition 7 increases the intensity of the liquid circulation (in the form of a tangential vortex motion with axial countercurrent).
  • the liquid medium with microorganisms or cells of animals or plants in the form of an upward flow rises and through the axial hole 8 of the partition 7 flows through the tangential radial channels 10 of this partition 7 down along the cylindrical side walls of the container 1.
  • the tangential shape of the radial channels 10 the specified annular partition 7 is not allowed to be fixed on its surface agglomerates of cells, which are discharged from the specified partition 7 by the fluid flow.
  • the gas aerates the cells of animals, plants or microorganisms on the surface of the liquid.
  • Example 4 A laboratory multiplatform gas-vortex bioreactor without an annular floating baffle 7 (Fig. 8) with an annular floating element 18 with removable blades 19 and needle elements 20 for surface bubbling (aeration) of the medium is being prepared for operation.
  • This version of the multi-platform gas-vortex bioreactor (platform No. 4) is used, for example, when working with microalgae in the mode of developing and optimizing the regulation of the cultivation process.
  • an annular version of the photobioreactor (with an annular zone 15) is used with a source 12 of intensive illumination of the culture and a video recording unit 13 of the cultivation process.
  • a seed dose of photosynthetic cells of microorganisms, such as chlorella is introduced (in the amount of 0.1 g per 1 liter of nutrient medium ), and turn on the rotation drive of the impeller 5, which supplies air enriched, for example, with carbon dioxide (for chlorella) through the pipe 3 tangentially into the container 1, and through the pipe 4 the exhaust gas.
  • the required number of revolutions of the impeller 5 of the fan is set.
  • a vacuum is created in the paraxial zone of the container 1 and an increased pressure on the periphery of this container 1.
  • a swirling gas flow is created above the annular surface of the cell suspension, which forms the culture liquid in the annular zone 15 turbulent rotational movement with mixing between the outer side of the cylindrical surface of the glass And and the container 1.
  • the annular floating element 18 with blades 19 rotates in the same direction and with the same angular velocity as the culture fluid and is kept afloat.
  • the blades 19 contribute to the mixing of the suspension, and the needle elements 20 contribute to a more intense surface bubbling of the culture liquid.
  • the gas aerates cells on the surface of the liquid. This provides optimal conditions for the cultivation of photosynthetic microorganisms.
  • the source 12 provides sufficient uniform illumination of the annular zone 15 of the chlorella suspension, the thickness of which is 30-50 mm.
  • the cultivation process is recorded by the video recording unit 13, which facilitates and accelerates the development and optimization of cultivation conditions.
  • Example 5 A multiplatform gas-vortex bioreactor is prepared for operation without an annular floating baffle 7 (Fig. 9, platform No. 5) with an annular floating element 18 with removable blades 19 (without needle elements 20 for surface bubbling).
  • the device works similarly to the description in example 4.
  • the annular floating element 18 with blades 19 rotates in the same direction and at the same angular velocity as the culture fluid and is kept afloat. Blades 19 provide more intensive mixing of the suspension.
  • the light source 12 provides sufficient uniform illumination of the annular zone 15 of the chlorella suspension. If necessary, the cultivation process is recorded by the video recording unit 13, which can be used for educational or scientific purposes.
  • This version of the bioreactor is used both in conjunction with bottom bubblers to create ascending aerating flows - airlift mode, and without them when working with media with a high protein content and, therefore, prone to abundant foaming.
  • the horizontal impeller 5 in addition to forming a gas vortex above the surface of the culture medium, functions as a mechanical defoamer.
  • Example 6 A laboratory multiplatform gas-vortex bioreactor without an annular floating baffle 7 (Fig. 10, platform No. 6) with an annular floating element 18 and needle elements 20 (aerators) for surface bubbling (without removable blades 19) is prepared for operation.
  • the device works similarly to the description in example 4.
  • the annular floating element 18 with blades 19 rotates in the same direction and at the same angular velocity as the culture fluid and is kept afloat. Needle elements 20 contribute to a more intense surface bubbling of the culture fluid.
  • the gas aerates the cells on the surface of the liquid. This provides optimal conditions for the cultivation of photosynthetic microorganisms.
  • the light source 12 provides sufficient uniform illumination of the annular zone 15 of the chlorella suspension. If necessary, the cultivation process is recorded by the video recording unit 13, which can also be used for educational or scientific purposes.
  • needle elements 20 perform two functions:
  • the claimed laboratory multiplatform bioreactor depending on the selected platform No. 1, 2, 3, 4, 5 or 6, provides the possibility of cultivating microorganisms, cell cultures or microalgae (lower phototropes) in the laboratory for research purposes, visually observe biotechnological processes and record them on a video camera, scale these processes, which confirms its versatility and thus ensures the claimed technical result.

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  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

L'invention concerne un bioréacteur à tourbillons de gaz à plateformes multiples pour la culture en suspension dans des conditions de laboratoire de micro-organismes, de cultures et de cellules végétales et animales et de micro-algues; ce bioréacteur comprend un récipient optiquement transparent (1) avec un couvercle, des conduits (3, 4), une roue à aubes horizontale (5), une cloison flottante annulaire (7), un dispositif d'aération et de mélange du milieu, un mécanisme de stabilisation (7) de la position de la cloison flottante annulaire par rapport à la surface de la suspension de cellules, une coupelle cylindrique creuse transparente et amovible (11) pour y placer une source d'éclairage et/ou une unité d0enregistrement vidéo, de mesure et de contrôle su processus industriel de culture. Le mécanisme de stabilisation (7) comprend un axe disposé coaxialement à la roue à aubes horizontale de manière à pouvoir être fixé sur la paroi d'extrémité (17) de la coupelle cylindrique (11). Le dispositif d'aération et de mélange du milieu comprend un élément flottant annulaire disposé dans le jour entre la paroi cylindrique du récipient et la cloison annulaire flottante. Au corps de l'élément annulaire flottant sont fixées des pales amovibles disposées radialement (19) et/ou des éléments en aiguilles formés régulièrement sur la surface d'extrémité supérieure de l'élément annulaire flottant en vue d'un barbotage de la surface du milieu nutritif liquide.
PCT/RU2021/000512 2021-01-12 2021-11-17 Bioréacteur à tourbillons de gaz à plateformes multiples de laboratoire WO2022154690A1 (fr)

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RU2021100420A RU2763318C1 (ru) 2021-01-12 2021-01-12 Лабораторный мультиплатформенный газовихревой биореактор

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CN115242951A (zh) * 2022-07-27 2022-10-25 国家海洋环境监测中心 一种基于图像扫描***的浮游动物快速检测方法

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RU2203938C1 (ru) * 2001-11-14 2003-05-10 Богданов Николай Иванович Установка для выращивания одноклеточных водорослей
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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115242951A (zh) * 2022-07-27 2022-10-25 国家海洋环境监测中心 一种基于图像扫描***的浮游动物快速检测方法

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