Classifications

• • 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
  • C12M23/00—Constructional details, e.g. recesses, hinges
  • C12M23/02—Form or structure of the vessel
  • C12M23/06—Tubular
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01D—SEPARATION
  • B01D19/00—Degasification of liquids
  • B01D19/0042—Degasification of liquids modifying the liquid flow
  • B01D19/0052—Degasification of liquids modifying the liquid flow in rotating vessels, vessels containing movable parts or in which centrifugal movement is caused
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01D—SEPARATION
  • B01D19/00—Degasification of liquids
  • B01D19/0042—Degasification of liquids modifying the liquid flow
  • B01D19/0052—Degasification of liquids modifying the liquid flow in rotating vessels, vessels containing movable parts or in which centrifugal movement is caused
  • B01D19/0057—Degasification of liquids modifying the liquid flow in rotating vessels, vessels containing movable parts or in which centrifugal movement is caused the centrifugal movement being caused by a vortex, e.g. using a cyclone, or by a tangential inlet
• • 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
  • C12M21/14—Bioreactors or fermenters specially adapted for specific uses for producing enzymes
• • 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
• • 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
  • C12M29/24—Recirculation of gas
• • 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
  • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
  • C12M41/48—Automatic or computerized control
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
  • Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
  • Y02E50/00—Technologies for the production of fuel of non-fossil origin
  • Y02E50/30—Fuel from waste, e.g. synthetic alcohol or diesel

Definitions

• the invention relates to biotechnology, namely, to methods and devices for cultivating microorganisms, including methylotrophic and methanotrophic bacteria, as well as their companions. More specifically, the invention relates to loop bioreactors and methods for their operation.
• a device for carrying out the fermentation process of methanotrophic bacteria which is a loop bioreactor, is known.
• the device contains horizontal and vertical sections made in the form of pipes forming a closed loop. In the upper part there is a cylindrical tank for gas separation. In this case, the diameter of the cylinder is significantly larger than the diameter of the pipes of the main circuit.
• Fluid circulation means are provided in the form of a pump.
• the plant is equipped with means for draining the culture liquid and exhaust gas, as well as inputs for the nutrient medium, titrating agent, and gases.
• the horizontal sections are equipped with static mixers to prevent bubbles from rising and coalescing.
• the degasser of this device is located in one place of the circuit, while the gas is supplied along the entire length. With an increase in the volume of the circuit, the volume of gas also increases, and the degassing section still remains in one zone. This leads to an increase in the volume of gas relative to the volume of bacteria and to high rates of movement of the mixture through the pipe (which is associated with additional losses).
• Known apparatus for growing microorganisms on natural gas (RU2738849C1).
• the apparatus is combined into a circulation circuit containing a jet aerator located vertically above a cylindrical barrel, a circulation means in the form of a pump, a vertical section with heat exchange jackets and mixers, a degassing tank, pipes for supplying and discharging liquid media and gases.
• the most important problem of the known design is to provide heat and mass transfer.
• heat removal from the installation is carried out in the pipeline part, while the main volume of the culture liquid and the main mass transfer, and, consequently, the main heat release, occur in the barrel.
• Due to the larger cross-sectional area of the barrel the velocity of the liquid is very low, resulting in a long residence time for the bacteria.
• Heat removal from the barrel is possible through additional internal heat exchangers, however, the placement of heat exchangers can change the flow regime and worsen mass transfer.
• the prior art fermenter for cultivating the biomass of methane-oxidizing microorganisms (RU2739528).
• the installation consists of two chambers, each of which contains reaction tubes, bubblers for introducing the gas phase, and nozzles located coaxially with the tubes.
• the nozzles are located under the pipes and directed upwards, deflectors are located above the pipes.
• the circulation in the apparatus is provided by a pump.
• the main disadvantage of the design is the length of the circulation section, which is less than a meter, which severely limits the conversion of methane, thereby worsening the comparative characteristics of the plant. To ensure the operation of the installation, a little more energy is required, so with a declared productivity of 4 g/l/h, which is 20% lower than the productivity achieved by the claimed invention, the energy consumption is 1 kWh/kg DIA.
• U-shaped fermenter including two vertical sections for downward and upward flows; U-shaped bend having a horizontal connecting section; an upper part in the form of a cylinder located above the U-shaped part and connecting the upper ends of the vertical sections; in this case, the diameter of the cylinder is significantly larger than the diameter of the pipes of the U-part.
• liquid circulation means are provided in the U-part of the fermenter. The withdrawal of the fermentation liquid is carried out through the outlet located in the upper part or horizontal connecting section of the U-shaped part of the fermenter.
• the fermenter is equipped with gas supply means, static mixing elements, water and nutrient salt supply means, temperature and pH sensors or analyzers, sensors for determining the concentrations of the components of the fermentation gas-liquid mixture, and pressure control devices.
• This device is characterized by the use of a container in the upper part as a degasser. Due to the shape of the container (“barrel”), the flow cross section changes, which leads to a decrease in the fluid velocity. This leads to the fact that, under the action of the Archimedean force, gas bubbles have time to rise to the surface and form a gas cavity from which gas can be removed, which reduces the efficiency of gas exchange in the circulating liquid.
• Significant disadvantages of this solution are problems with barrel scaling, the presence of a low mass transfer area in which bacteria reside for a long time, the presence of a significant volume filled with methane-air mixture, and the relatively high metal consumption of the structure.
• the fermenter includes four blocks interconnected to form a closed loop for the movement of the cultural liquid (CL), a pump connected to the closed loop, bubblers, mixers, supply means components of QOL, selection of QOL, removal of gases, supply and removal of coolant, means of measuring QOL parameters.
• the first and third blocks are made vertically oriented, they include a pipe made with the possibility of thermostatting the CL, the second and fourth blocks are made horizontally oriented, they include containers made with the possibility of degassing the CL.
• the fermentation plant includes the specified fermenter, as well as lines for water treatment, preparation of a nutrient medium, preparation of a titrating agent, preparation of seed material, preparation of a gas medium, purification of off-gases from CO2, separation, sterilization and drying.
• the degassing tanks used in this installation are characterized by a change in the direction of the CL flow when it passes through the degasser tank.
• the gas bubbles stick together and stray to one side, forming a volume with a low mass transfer, which reduces the efficiency of the installation.
• a certain amount of energy is spent on turning the flow, additional resistance is created, which also negatively affects the performance of the installation.
• the volume of the degasser grows faster than the useful volume of the fermenter, which makes it difficult to scale up the plant industrially.
• the technical problem solved by the claimed invention lies in the need to overcome the above disadvantages inherent in analogues and the prototype by creating a scalable, energy-efficient, highly productive industrial fermenter and fermentation unit.
• the technical result achieved by using the claimed invention is to increase the rate and efficiency of gas exchange in the process of circulating the culture liquid along the fermenter circuit while making it possible to scale the fermenter and the fermentation plant as a whole to industrial volumes of more than 150 m 3 .
• the technical advantage of the claimed invention is also the provision of a methane conversion of more than 65%, as well as a reduction in the energy consumption of the fermenter and the installation as a whole in industrial use due to the modification of the degassing and mixing units of the fermenter.
• the fermenter is also characterized by the possibility of carrying out service maintenance without stopping the technical process.
• At least one bubbler located in the lower or upper part of the vertically oriented block, installed after the pump in the direction of flow along the circuit, at the inlet for supplying gaseous media to the first or third,
• the degassing block is installed in the part of the vertically oriented block opposite from the bubbler to ensure that the direction of movement of the CL flow is maintained when passing through the degassing block, while this block includes at least one once-through degasser that provides degassing of the flow passing through the cross-section area up to 400 square meters. cm., and static mixers are installed along the length of a vertically oriented block between the bubbler and the degassing unit to provide the possibility of forming gas bubbles in the CL flow with a diameter of up to 5 mm at a distance of not more than 45 cm from each installed mixer in the direction of the flow.
• the horizontally oriented blocks of this fermenter can be formed by two interlocking 90° bends from the vertically oriented blocks, while the length of the horizontally oriented block is not less than the diameter of the pipeline of the vertically oriented block.
• the degassing unit may contain at least two direct-flow degassers installed in parallel between the inlet and outlet adapters, which ensure the distribution of the incoming flow over the degassers and the collection of outflows from the degassers, respectively.
• the degasser can be made in the form of a flow-through cylindrical body having end inlet and outlet for the flow of liquid and degassed medium, respectively, inlet and outlet screws installed in the case coaxially to each other near the inlet and outlet, respectively, located from each other.
• the input screw of such a degasser is a central cylindrical sleeve, on the outer side surface of which evenly spaced blades are fixed, each of which is a curved surface, its input the edges are made straight and oriented radially to the hub axis, the trailing and side edges are curved, while the angle a between the hub axis and the tangent drawn to the blade at each point of the trailing edge is determined in accordance with the following relationship: where the dimensionless coefficient K is equal to 4-7, r is the distance from the point of measurement of the angle a to the axis of the bushing, R is a constant value characterizing the distance from the axis of the bushing to the outer side edge, while the diameter of the central bushing is 0.1–0.5 from input screw diameter equal to 2R.
• the distance between the inlet and outlet screws of the degasser is determined by the following
• L is the distance between the screws
• N is 10 ⁇ -15
• R is the radius of the degasser body
• angle a is the angle between the axis of the input screw and the tangent to the blade at each point of the trailing edge of the input screw.
• a static mixer can be a flow-through cylindrical body, inside which blades are fixedly mounted around a cylindrical sleeve, the leading edges of which are straight and oriented radially to the axis of the sleeve, and the trailing edges are made in the form of a zigzag and have a length that is more than 2 times greater than the length of the leading edge , while the blades are completely described by the following parametric equation in cylindrical coordinates: where z is the distance from the leading edge to a given point of the blade surface, 0 ⁇ z ⁇ z m , z m is the height of the mixer (corresponds to the distance from the leading edge to the trailing edge of the blades), Rhub > bushing radius, r is the distance from the bushing axis to this point blade surface, R ⁇ ub ⁇ r ⁇ R, where R is the radius of the mixer body.
• the fermenter pump circulates the liquid around the circuit at a speed of 0.8 to 1.5 m/s.
• the bubbler for supplying gaseous media is made with the possibility of forming gas bubbles in the CL with a diameter of not more than 5 mm.
• Each vertically oriented block can be made of pipe sections with a heat exchange jacket, interconnected through static mixers, while a vertically oriented section with a heat exchange jacket has a diameter of 200 to 1000 mm and a length of 200 to 1000 mm.
• Means for supplying CL components are at least one inlet or branch pipe for supplying liquid media to the main circuit with CL; at least one inlet or branch pipe for supplying gaseous media, located in the block with the bubbler.
• the means for removing residual and gases formed during the life of microorganisms represent at least one outlet or branch pipe located in the unit with the degasser.
• the means of selection of QOL is at least one outlet or branch pipe located in a horizontally oriented block.
• Means for measuring QOL parameters are means for measuring temperature, including temperature sensors associated with the control controller; means for measuring the pH of the medium, including pH sensors associated with the control controller; means for measuring the composition of gases, including sensors for measuring the concentrations of outgoing gaseous media associated with the control controller; sensors for measuring the flow of incoming gaseous media, including gas flow meters associated with the control controller; means for measuring the liquid flow through the fermenter, including a flow meter for the liquid flowing out of the fermenter, connected to the control controller; means for measuring the chemical composition of CL, including sensors for the concentration of dissolved oxygen associated with the control controller; means for measuring the volume of liquid in the fermenter, including a level sensor connected to the control controller; means for measuring the density of QOL, including an automatic optical flow cell, the
• the control controller based on the readings of the means for measuring QOL parameters, their combination, as well as data on the position of the valves and the frequencies of the pumps, within the framework of the execution of the specified algorithms, issues commands to the valves, pumps and system elements, thereby regulating the fermentation parameters and automatically maintaining the temperature, pH, gas flow rate, QOL volume, dissolved oxygen level, pressure and QOL level in the separator according to the setpoint set by the algorithm or the operator.
• the fermenter may include two degassing units and two bubblers, each of the vertically oriented units being provided with a bubbler and a degassing unit.
• the claimed technical result is also achieved by the solution of the fermentation installation, including a fermenter made according to claim 1 of the claims, as well as the following devices connected in a technological sequence to ensure a closed cycle for the cultivation of methanotrophic microorganisms:
• the outlet from the fermenter, intended for the selection of QOL is connected to the concentration line, which in turn is connected to the sterilizer, which turns into a spray dryer,
• the water treatment line includes a filtration plant that provides treatment of incoming water to bring its quality in line with technological requirements, including mechanical cleaning, biological treatment, and desalination.
• the nutrient medium preparation line is configured to supply a nutrient medium of a given concentration prepared from a concentrate or individual components automatically or manually.
• the titrating agent preparation line includes a container configured to thermostat the prepared titrant solution and equipped with a stirrer.
• the gas medium preparation line consists of two sub-lines - methane and air, and includes compressors, devices for preparing natural gas, air to obtain a gas or gas mixture of the required quality in accordance with technological requirements, including the removal of water and unwanted impurities.
• the seed preparation line includes at least one fermenter.
• the concentration line includes a membrane filtration unit and a centrifuge for obtaining a paste with a moisture content of 77-80%, as well as a container for collecting the filtrate.
• the sterilizer provides sterilization of the culture liquid concentrate at 90-99 °C.
• the drying line includes a spray dryer.
• the gas outlet line provides the possibility of safe removal of residual gases and gases formed during the life of microorganisms and contains a pressure control valve in the fermenter, a dryer and a gas composition sensor.
• the plant also includes a separator connected to the degassing unit and designed for complete separation of the gas-liquid mixture leaving the degassing unit of the fermenter.
• the main volume of the fermenter is a closed predominantly vertically oriented pipe circuit, due to which there are no gas cavities and a constant interphase boundary, and high gas and mass transfer can be achieved due to regular crushing of bubbles by static mixers.
• the interfacial boundary is maintained in a separate separator, which can be replaced and sent for washing without stopping the production process and stopping the circulation of CL in the fermenter.
• the volume of the gas cavity in the separator of the proposed installation is insignificant compared to the volume of the degassing tank of the prototype.
• Horizontally oriented blocks of the inventive fermenter can be reduced to small sizes, as a result of which zones without gas are removed, that is, with low mass transfer.
• reducing the length of the horizontal blocks to 10% of the total length of the fermenter circuit improves the energy efficiency of the plant and reduces the area of its installation.
• the scalability of the plant is achieved, among other things, by overcoming structural problems associated with mass transfer, heat transfer, gas removal and pumping.
• FIG. 1 schematically shows the design of the inventive fermenter
• Fig. 2 schematically shows the flow diagram of an example of the implementation of the proposed fermentation plant
• Fig. 3 is a schematic representation of an implementation example of a degasser
• FIG. 4 shows an example of the implementation of a static mixer
• figure 5 shows a degassing unit, consisting of several parallel degassers
• figure 6 shows a graph of the distribution of dissolved oxygen along the circuit of the fermenter, the reading is from the pump.
• figure 7 shows a variant of the design of the fermenter with horizontal blocks formed by 90° bends
• figure 8 schematically shows the installation of several static mixers in the circuit when it is scaled.
• the inventive fermenter allows you to change the mode of cultivation in a controlled manner, for example, replace the process with a maximum growth rate and low bacterial density with a process with a low growth rate and maximum bacterial density. In this way, the nature of substrate consumption, the composition of biomass, the power of heat release, electricity consumption and other parameters can be changed.
• the claimed invention may be subject to various changes and modifications, clear to a specialist on the basis of the present description. Such changes do not limit the scope of the asserted claims.
• the fermentation plant of the proposed design is characterized by a balance of operational parameters, such as substrate conversion, fermenter productivity, its energy efficiency, and allows their flexible variation in a wide range without fundamental changes in the design.
• the fermenter includes at least four blocks interconnected to form a closed circuit for the movement of the culture fluid (CL), while the circuit is formed using alternately arranged horizontally and vertically oriented blocks.
• the first 7 and third 9 blocks are made predominantly vertically oriented.
• Each of these blocks includes at least one section (pipe) for QOL 4, made with the possibility of temperature control of QOL.
• said vertically oriented blocks can be made of several identical sections 4 (preferably, each vertically oriented block is made of seven sections covered with a heat exchange jacket), between which static mixers are installed, which also ensures uniform arrangement of mixers in vertically oriented blocks.
• the number of sections is generally determined based on the number of mixers and heat exchangers, as well as the convenience of their maintenance.
• the section with a heat exchange jacket has a diameter of 200 to 1000 mm and a length of 200 to 1000 mm.
• the second 8 - top, and the fourth 10 - bottom blocks are made mainly horizontally oriented.
• the blocks are connected to each other in a single closed circuit by means of L-shaped branches 26.
• the diameter of the branches coincides with the diameters of the second 8 and fourth 10 horizontal blocks, and with the first 7 and third 9 vertical blocks, the branches 26 are connected through confusers.
• the first and third vertical blocks are pipes with a diameter of 200 mm to 1000 mm, a length of 400 mm to 1000 mm.
• the length of the horizontal blocks 2 and 4 can be reduced up to a complete reduction. So, in one of the embodiments, horizontally oriented blocks can be formed by conjugated branches from vertically oriented blocks.
• the size of the pipes that make up the circuit is determined by the required working volume of the fermenter, and for a working volume of, for example, 50 m 3 , it can reach several tens of meters, while the height of the pipes can be 20 m or more.
• the dimensions of such a fermenter can reach 10 m in width and 25 m in height.
• the pump 1 is located at the inlet to the first block 7 and provides pumping of the QOL flow at a speed of 0.8 to 1.5 m/s.
• the number of pumps can be changed if it is necessary to increase the volume of pumped liquid.
• the design of the fermenter may include two bubblers to increase the length of the circuit, installed in different vertically oriented blocks.
• one bubbler is in the lower part of one block (at the flow inlet to this block), and the second is in the upper part of the other (also at the inlet of the QOL flow into this block).
• a degassing block 5 is installed, the design of which provides for the possibility of maintaining the direction of the CL flow when passing through the degasser.
• the QOL flow at the inlet and outlet of the degasser does not turn, but flows straight. Accordingly, in the presence of two bubblers, two degassing units are installed.
• the degassing unit is installed coaxially with the pipe of the vertically oriented unit. Each degassing unit is connected to a separator tank that completely separates the gas-liquid mixture leaving the degasser, installed nearby on the floor or frame.
• the degassing unit can be made of one flow-through cylindrical body having an end inlet for supplying the degassing medium and an end outlet for the degassed medium.
• the casing diameter is selected taking into account the diameter of the pipeline, in which the degasser is supposed to be installed and provides degassing of the flow with a cross-sectional area of up to 400 sq. see.
• the constructive solution of the degasser can be any available with the provision of degassing of the flow passing through the cross section of the degasser with an area of up to 400 square meters. cm without changing the flow direction.
• One of the variants of the degasser can be a solution that consists in installing a stationary screw near the inlet to the degasser body, without the possibility of rotation or translational movement of the inlet screw, and at a certain distance from the inlet screw in a cylindrical body - the outlet screw, which ensures straightening of the gas-liquid flow.
• the input and output screws are installed coaxially to each other.
• the output screw can be placed permanently or with the possibility of translational movement along the cylindrical body to select the optimal point for its placement relative to the input screw.
• the inlet screw of the degasser is a central cylindrical bushing, on the outer side surface of which uniformly spaced blades are fixed, each of which is a curved surface, its leading edges are made straight and oriented radially to the bushing axis, the outlet and side edges are made curvilinear, while the angle a between the hub axis and the tangent drawn to the blade at each trailing edge point, is determined in accordance with the following relation (1): where the dimensionless coefficient K is equal to 4-7, r is the distance from the point of measurement of the angle a to the axis of the sleeve, R is a constant value characterizing the distance from the axis of the sleeve to the outer side edge, while the diameter of the central sleeve is 0.1-0.5 from input screw diameter equal to 2R.
• the distance between the screws is chosen to ensure that at least 90% of the gas bubbles are drawn into the vortex air cord formed during the passage of the liquid medium flow through the inlet screw blades towards the outlet screw.
• the distance between the screws L is chosen such that the length of the air cord formed between the input and output screws is sufficient to draw the vast majority of the bubbles into the cord.
• the distance between the screws depends on the diameter of the body of the degassing device and can be determined from the relation (2): where L is the distance between the screws, the coefficient N is equal to 10 ⁇ -15, R is the radius of the device body (equal to the distance R from the axis of the input screw hub to the outer side edge of the input screw), angle a is the angle between the axis of the input screw hub and the tangent to blades at each point of the trailing edge of the input screw.
• the screw is installed in such a way that the outer side edges of its blades are conjugated with the inner cylindrical surface of the device body, and the screw axis coincides with the axis of the degasser.
• the degassing unit is formed by several degassers installed in parallel, having an identical design, for example, as described above, combined adapters 47 and 48, which ensure the distribution of the inlet flow over the bodies of individual degassers and, accordingly, the collection of degassed flows into a single stream.
• This approach makes it possible to maintain the length of the degassing unit while increasing the diameter of the pipes of the main circuit.
• the volume of the degassing block increases in proportion to the total volume of the fermenter by increasing the total cross-sectional area.
• At least one static mixer is installed along a vertically oriented block.
• static mixers are installed at a distance from each other along the entire length of the section between the bubbler and the degasser, so that bubbles up to 5 mm in size are formed at a distance of up to 45 cm from each mixer.
• Static mixers can have various design and layout options. Both well-known designs of Sulzer SMV, SPX FLOW Lightnin Inliner and original ones can be used. Static mixers create the main hydrodynamic resistance when pumping CL, since they create turbulence and provide a velocity gradient necessary for mixing and crushing bubbles. Thus, the main requirement for choosing a mixer is to maintain the quality of mixing and mass transfer, which makes it possible to obtain a productivity of 5 g/l/h using air, with a total energy efficiency of 1 kW*h/kg DIA.
• the mixer can be made in the form of an open body with a radius R and a height z m , inside which a central, predominantly cylindrical bushing is installed, along the side surface of which fixed blades are fixed.
• Each blade is a complex curved curvilinear plate bounded by four edges - inlet and outlet (in the direction of flow movement), as well as outer and inner side edges, conjugated, respectively, with the inner side surface of the housing and the outer side cylindrical surface of the central sleeve.
• the input edges are made straight, oriented radially and have a length equal to the difference between the radii of the mixer body and the bushing.
• the inner side edge is also made straight and is oriented along the side surface of the central sleeve coaxially with its axis.
• the blades are completely described by the following parametric equation in cylindrical coordinates: where z is the distance from the leading edge to a given point of the blade surface, 0 ⁇ z ⁇ z m , z m is the height of the mixer (corresponds to the distance from the leading edge to the trailing edge of the blades), Rhub > bushing radius, r is the distance from the bushing axis to this point blade surface, R bU b ⁇ r ⁇ R, where R is the radius of the mixer body.
• a version of the static mixer is shown in figure 4.
• the body of the mixer can be made hexagonal, which makes it possible to fill the section of a large diameter pipeline with mixers such as honeycombs (fig. 8).
• Mixer blades with such a housing can also be described in accordance with the above data.
• the design of mixers allows you to change the geometry of the blades to optimize the size, thereby increasing the useful section.
• the pressure drop across one section with mixers does not depend on the cross-sectional area.
• Scaling the fermenter implies changing the length of the outline and/or the cross-sectional area of its outline.
• the length of the fermenter circuit is determined by the rate of dissolution of gaseous substances in water. As the gas bubbles move, the concentration of nutrients substances, therefore, and the rate of their dissolution falls. When the dissolution rate becomes critically low, the depleted gas residues are taken off.
• the length of the passage of the gas portion determines the length of the fermenter and is fundamentally limited. There is profiling along the length of the contour. The sequential arrangement of several gas inlet and outlet points is impractical, as it excessively increases the height of the installation. Thus, an increase in the volume of the fermenter circuit is achieved by increasing the diameter of the pipes that make it up, that is, the cross-sectional area of the pipes of the fermenter circuit.
• the length (extension) L of the degassing block of the fermenter is proportional to the radius R of its body to the power of 3/2: z L-R2
• volume V of the degassing unit is proportional to 7 V-R2
• V nNLR 2
• the length of the degassing block remains fixed, and the volume grows in proportion to the cube of the linear size.
• the flow regime in each individual section remains unchanged.
• the design of the fermenter means that there are no stagnant zones or zones with increased residence time, so there is no risk of a critical temperature or concentration gradient.
• the inventive fermenter also contains means for supplying QOL components 13, selecting QOL 14, supplying gases 11, removing residual gases (exhaust gases) 12 formed during the life of microorganisms, supplying and removing coolant 16, means for measuring QOL parameters 17, 18, 19, 20, 21, 22.
• temperature measuring means 18 can be used, including temperature sensors associated with the control controller; means for measuring the pH of the environment 17, including pH sensors associated with the control controller; means for measuring the composition of gases 22, including sensors for measuring the concentrations of outgoing gaseous media associated with the control controller; sensors for measuring the flow of incoming gaseous media 23, including gas flow meters associated with the control controller; means for measuring the flow of liquid through the fermenter, including the flow meter flowing from the fermenter liquid 24 associated with the control controller; means for measuring the chemical composition of QOL, including sensors for the concentration of dissolved oxygen 20 associated with the control controller; means for measuring the volume of liquid in the fermenter, including a level sensor 19 associated with the control controller; means for measuring the density of QOL, including an automatic optical flow cell 25, the measurements of which are transmitted to the control controller; means for measuring pressure, including pressure sensors 21 associated with the control controller.
• At least one inlet or branch pipe for supplying liquid media 13 to the main loop with QOL can be used; at least one inlet or branch pipe for supplying gaseous media 11 located in the block with the bubbler.
• At least one outlet or branch pipe located in the fourth horizontally oriented block 10 is used as a means of selection of QOL 14.
• the inventive fermentation plant for continuous cultivation of microorganisms includes the following devices, connected in a technological sequence to ensure a closed cycle of cultivation of microorganisms: a fermenter of the claimed design, a water treatment line 35, including a filtration plant that provides treatment of incoming water to bring its quality in line with technological requirements, including mechanical cleaning, biological treatment, desalination.
• Line 35 is connected to a culture medium preparation line 37 and to a titrating agent preparation line 38, the outlets of which are connected to the respective fermenter inlets and to the seed preparation line 40; the gaseous medium preparation line 36, 39, which is connected to the corresponding inlet of the fermenter and the seed preparation line; 44, as well as the gas outlet line 45 and the temperature control circuit 46.
• the nutrient medium preparation line 37 has the ability to supply a nutrient medium of a given concentration, prepared from a concentrate or individual components automatically or manually.
• Seed preparation line 40 includes at least one fermenter.
• the titrating agent preparation line 38 includes a container configured to thermostat the prepared titrant solution and equipped with a stirrer.
• the gaseous medium preparation line consists of two sub-lines - methane 39 and air 36, and includes compressors, devices for preparing natural gas, air to produce gas or a gas mixture of the required quality in accordance with technological requirements, including the removal of water and unwanted impurities.
• the concentration line includes a 41 membrane filtration unit and a 42 centrifuge to obtain a paste with a moisture content of 77-80%, as well as a collection tank filtrate.
• the culture liquid concentrate is sterilized at 90-99 °C in the 43 sterilizer.
• the drying line includes a 44 spray dryer.
• the gas outlet line 45 allows for the safe disposal or recycling of residual and microbial gas, and contains a pressure control valve in the fermenter, a dryer and a gas composition sensor.
• the temperature control circuit 46 has the ability to maintain the specified temperature of the heat carrier when it is fed into the fermenter.
• the heat exchanger is a heat exchange jacket installed outside the pipe with CL.
• the coolant enters the jacket from the collector and flows through the spiral channels.
• the flow channel goes inside the fermenter. Inside, the channel changes shape or cross section to increase the heat exchange area.
• the optimal geometry of the heat exchange circuit is selected based on the heat release power of the CL, which depends on the cultivation mode.
• Inlet and outlet pipes and fittings are divided into two categories.
• the first category is rigidly attached to structural elements and cannot be moved along the contour. It includes a branch pipe for supplying incoming gases, outgoing gases, inlets and outlets of the coolant to the heat exchanger.
• the second category can be installed in an arbitrary place or based on the following considerations. Several titrating agent inlets are distributed along the length of the circuit as evenly as possible.
• the pH sensor is not recommended to be installed near the inlet of the titrating agent.
• An air outlet should be installed in the upper horizontal section, a fitting for the selection of QOL in the degassed zone.
• the fermenter has pressure and dissolved oxygen profiling by distance from the pump or bubbler. Profiling must be taken into account when interpreting the results obtained from the corresponding sensors, depending on the point of their installation. Additional pipes or fittings can also be built in, for example, a separate valve for sampling QOL for analysis.
• the inventive fermenter and fermentation unit operate as follows.
• the preparation of a nutrient medium for circulation along the fermenter circuit occurs by dissolving basic salts and trace elements in water. from the water treatment line, as well as by mixing the resulting solution with the filtrate from the separation line.
• a solution of basic salts for example, MgSO4 7H2O, KC1 and K2SO4
• a solution of trace elements for example, CuSO4 5H2O, FeSO4 7H2O, MnSO4 5H2O, NZVOZ, ZnSO4 7H2O, NiSO4 7H2O, CoSO4 7H2O, Na2MoO4 2H2O, H3PO4 - 85% solution
• trace elements for example, CuSO4 5H2O, FeSO4 7H2O, MnSO4 5H2O, NZVOZ, ZnSO4 7H2O, NiSO4 7H2O, CoSO4 7H2O, Na2MoO4 2H2O, H3PO4 - 85% solution
• the volume of containers and concentrations of solutions are determined by the requirements of the technological regulations, depending on the stages of the technological process and the microorganisms used.
• the inoculum is prepared by successive growth of the cell suspension using the same nutrient medium, natural gas and oxygen; under the same conditions as the main cultivation process: with stirring, thermostating, constant pH.
• the nutrient medium for seeding bioreactors is supplied from the PS preparation line, natural gas and air are supplied from the HS preparation line, and the titrating agent from the TA preparation line.
• the inoculum is a cell suspension grown in an Erlenmeyer flask.
• the cell suspension obtained in the bioreactor with the largest volume on the PS preparation line is fed to the fermentation line.
• the fermenter made in accordance with the previously described design features, is filled with a nutrient medium coming from the PS preparation line up to 97.5% of the OF. Then the instrumentation system is turned on and the QOL circulation is started with the built-in pump. Next, the temperature and pH of the medium are set to the operating values provided for by the technological regulations, the pH of the medium is maintained by introducing a titrant from the TA preparation line. Inoculation material from the PS preparation line is introduced into the nutrient medium in the form of a bacterial suspension of methanotrophic bacteria in the amount of 2.5% OF with a concentration of about 18 g/l, so that the starting concentration of bacteria in the fermenter reaches about 0.45 g/l. After that, the HS supply begins through the corresponding inlets (bubblers). As a gas mixture, as a rule, a mixture is used natural gas (methane) and air. Air and natural gas are fed into the fermenter separately.
• a gas mixture as a rule, a mixture is used
• the flow of culture fluid circulates in a closed circuit of the fermenter under the action of a centrifugal pump 1 in the direction from the first to the fourth block.
• Air and methane are fed through the gas treatment lines in separate streams to the bubbler.
• the gas flows are distributed equally.
• the gas passes into the culture liquid in the form of bubbles up to 5 mm in diameter.
• the total volume fraction of the gas phase in the fermenter varies from 0 to 12%.
• static mixers create a velocity gradient along the length of the first and third blocks, crushing the bubbles as they move with the flow.
• the distance between the static mixers is chosen to maintain the bubble diameter in the range of 0.1-5 mm.
• Dissolved methane and oxygen are the nutrient substrate.
• the rate of assimilation by bacteria is much higher than the rate of transition from the gas phase to the liquid phase, where the substance flux depends on the area of the interfacial surface and the concentration profile. Based on this, the characteristics of the mixers are optimized for the most complete and uniform dissolution of the gas along the length of the fermenter. Further, bubbles depleted in methane, oxygen and saturated with carbon dioxide must be removed from the volume of the fermenter. This function is provided by a direct-flow degasser.
• the flow running on the inlet screw of the degasser, twists, acquiring a velocity profile corresponding to the vortex motion.
• a centrifugal force arises, creating a pressure gradient, under the influence of which the liquid, as a denser fraction, collects at the pipe wall, and the bubbles accumulate in the center in the region of the lowest pressure, merging into a gas cord.
• a tube is connected to the gas cord, through which the gas is removed from the fermenter and enters the separator. This solution allows you to capture more than 95% of the gas. After passing through the degasser, the culture liquid enters the centrifugal pump and the cycle starts over.
• the device of the degasser allows operation in modes with different ratios of gas and liquid flows, as well as the volume fraction of gas in the fermenter. Depending on the selected mode, the degassing efficiency and the amount of liquid taken from the fermenter along with the flow of the outgoing gas change.
• the device allows while maintaining the quality of degassing above 95%, capture significantly less culture fluid than is required to ensure the flow.
• a mode is selected in which the outflow contains a significant amount of liquid.
• the gas-liquid mixture passes through a separator, which has a much smaller volume than the fermenter.
• the culture liquid descends to the bottom of the separator and is pumped back to the fermenter.
• the liquid level in the separator is kept constant, and the flow rate of the transfer pump is an important parameter characterizing the cultivation process.
• the purified gas enters the outlet gas line.
• the fermentation unit operates in the continuous cultivation mode with a flow.
• the nutrient medium containing dissolved mineral salts is fed through several injection points distributed along the fermenter circuit. Excess QOL enters the concentration line for further processing and isolation of dry matter.
• a titrating agent is supplied through several distributed injection points, which also contains a source of nitrogen nutrition.
• the nutrient medium is prepared by dissolving the main trace elements and salts in water, resulting in a concentrate.
• the concentrate is automatically mixed with water coming from the water treatment line or filtrate from the concentration line and pumped into the fermenter in a proportion set algorithmically. All initial chemical components can be class "T" in accordance with GOST 13867-68.
• Culture fluid with a dry weight content of 1-3% is concentrated in two stages, first by membrane filtration, then by centrifugation to 20-23% dry weight.
• the separated filtrate is an aqueous solution of residual PS components and soluble products of bacterial metabolism.
• the concentrate aged in the sterilization unit is sent to the dryer. Drying of biomass is carried out by spraying through a nozzle into streams of hot dry air. The dry matter thus obtained can be used as a final product or sent for granulation.
• Satellites are able to utilize the products of metabolism and lysis, as well as to isolate their own metabolites, such as B vitamins, which stimulate the growth of M. capsulatus.
• the main satellites are Cupriavidus gilardii, Thermomonas hydrothermalis, Brevibacillus fluminis, Paenibacillus lactis
• the plant volume of 378 l with a productivity of more than 5.5 g/l/h makes it possible to obtain 2079 gASV/h.
• the total height of the installation is approximately 5.8 m, the internal diameter of the main (vertical) section is 212 mm.
• a configuration was used with the installation of one degassing unit and one bubbler.
• the diameter of the degasser and horizontal blocks is 153 mm.
• the centrifugal pump circulates the culture liquid at a speed of 1.2 m/s.
• the main circuit and sections (in the amount of 7 pieces) of the fermenter are made of stainless steel, which is inert to the components of the QOL and washing liquids.
• the installation can withstand 6 atm. excess pressure. Static mixers of our own design in the amount of 15 pieces installed in the first, second and third blocks were used as mixers.
• FIG. 6 shows a graph of the distribution of the level of dissolved oxygen along the length of the fermenter circuit with a reading from the pump. From the graph it can be seen that for most of the length the level is close to average and can be averaged to a plateau, which is a consequence of the fight against profiling by optimizing the location and geometry of the mixers. On the right side of the graph, one can see a steady decrease in the concentration of dissolved oxygen, which indicates a critically decreasing gas concentration in the bubbles. If the concentration falls below the acceptable level, the gas must be renewed.
• the amount of micro and macro elements is supplied in accordance with the need of microorganisms for the synthesis of biomass, calculated on the basis of the elemental composition of DIA.
• the composition of the concentrate includes the following substances in g/l:
• Air and natural gas with a methane content of about 95% were supplied as gas.
• the pH of the medium was maintained at around 5.6 by introducing a 5% ammonia solution as a titrating agent.
• the culture liquid was maintained at a temperature of 42°C.
• the working pressure is 3.3 atm. izb. Energy efficiency was 1 kWh per kilogram of DIA.

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