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
  • C12M23/58—Reaction vessels connected in series or in parallel
• • C—CHEMISTRY; METALLURGY
  • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
  • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
  • C02F3/00—Biological treatment of water, waste water, or sewage
  • C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
  • C02F3/348—Biological treatment of water, waste water, or sewage characterised by the microorganisms used characterised by the way or the form in which the microorganisms are added or dosed
• • 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
  • C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
  • C12M33/22—Settling tanks; Sedimentation by gravity
• • 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
  • C12M37/00—Means for sterilizing, maintaining sterile conditions or avoiding chemical or biological contamination
• • C—CHEMISTRY; METALLURGY
  • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
  • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
  • C02F2101/00—Nature of the contaminant
  • C02F2101/30—Organic compounds
  • C02F2101/306—Pesticides
• • C—CHEMISTRY; METALLURGY
  • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
  • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
  • C02F2101/00—Nature of the contaminant
  • C02F2101/30—Organic compounds
  • C02F2101/34—Organic compounds containing oxygen
• • C—CHEMISTRY; METALLURGY
  • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
  • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
  • C02F2101/00—Nature of the contaminant
  • C02F2101/30—Organic compounds
  • C02F2101/36—Organic compounds containing halogen
  • C02F2101/363—PCB's; PCP's
• • C—CHEMISTRY; METALLURGY
  • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
  • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
  • C02F2101/00—Nature of the contaminant
  • C02F2101/30—Organic compounds
  • C02F2101/36—Organic compounds containing halogen
  • C02F2101/366—Dioxine; Furan
• • C—CHEMISTRY; METALLURGY
  • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
  • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
  • C02F3/00—Biological treatment of water, waste water, or sewage
• • 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 present invention relates to a method and a device for culturing living cells by coupling a bioreactor container with a selection automaton.
• Waste treatment is an increasingly constant concern of citizens and governments. A significant part of the treatments is carried out in factories using tanks inoculated with a bacterial flora. But the bacterial flora evolves over time and this evolution is often unfavorable to the reactions that one wishes to see intervening in the tanks.
• the problem of drift of bacterial cultures is a general problem that is found for example in the pharmaceutical industry. In this industry, drift is avoided by operating in a sterile manner. But it is inconceivable economically to work in such conditions for example in a waste treatment plant.
• the device described therefore behaves like a process automated selection which at the same time eliminates static variants of living cells, that is to say living cells which stagnate in pipes and containers and favors dynamic variants remaining in suspension, which are better and better adapted to growing conditions.
• biodegradation of wastewater implements to date diverse bacterial populations whose nature escapes the control of the operator, and without it being possible to use only the species selected for their performance or their effectiveness with respect to the substrate, that is to say to the compounds to be degraded present in the waste water. It would therefore be desirable to have a technique, in particular of biodegradation of wastewater, which makes it possible to use essentially the species selected for their performance or their effectiveness with respect to the compounds to be degraded present in the wastewater.
• the Applicant has discovered a process making it possible to reproduce and control the proliferation conditions in a large bioreactor container, or in a natural environment such as a lagoon or a body of water for example, and operating continuously, semi-continuously or discontinuously, without resorting to containment and sterilization, and by privileging the dynamic variants of living cells, which are better and better adapted to the culture conditions.
• This process is essentially based on cooperation between a bioreactor container and an automatic selection device for living cells.
• the present application relates to a continuous, semi-continuous or discontinuous treatment process for a substrate, in which said substrate installed in a bioreactor container is subjected to the action of a culture of living cells C1 making it possible to carry out a reaction R1 on said substrate and in which the medium is inoculated periodically and preferably regularly with the aid of living cells C2 improving said reaction, said living cells C2 being obtained from a selection carried out by a selection device automatic, preferably exclusively in suspension, of a population of dynamic living cells and said automatic living cell selection device being fed either by a different substrate or by the same substrate as the bioreactor container and being originally inoculated by the living cells C1 present in the tank of the bioreactor container, and in which advantageously o n takes living cells from the tank of the bioreactor container to transfer them to the automatic selection device.
• living cells can be added to the selection device or to the bioreactor container, if desired, at any time, for example to increase cell concentration or introduce new species.
• living dynamic cells designates living cells proliferating in suspension and subjected to a directed selection (unlike “static living cells”, designating living cells adhering to the surface containers and pipes, thus escaping selection).
• the static living cells are advantageously eliminated periodically.
• living cells proliferating in suspension and subjected to a directed selection will be used as living cells C2.
• C2 will be used as living cells, not dynamic living cells, but static living cells.
• the substrate allows the maintenance of cultures of living cells C1, C2, etc.
• the device for automatic selection of dynamic living cells comprises: two or more receptacles making it possible to receive and maintain cultures of living cells in suspension, a set of means making it possible to feed separately these containers in sterilization, cleaning or neutralization fluids a set of means for supplying these containers with gas a set of means for supplying these containers with substrate a set of means for transferring the contents of a container in the other and vice versa - a set of means making it possible to evacuate all or part of the content of these containers to another device such as a bioreactor container a set of means making it possible to evacuate all or part of the content of these containers to a trash can.
• living cells C1 are present in the bioreactor container tank and in the automatic selection device. Over time, the selection device favors (selects) the appearance and proliferation of dynamic living C2 cell variants, always better adapted to the culture conditions and counter-selected the less well adapted living C1 cells.
• the living cells C2 are transferred to the bioreactor container where they compete with the living cells C1 and then supplant them. At the end, it can be seen that the population of living cells C1 has been replaced by living cells C2.
• living cells are taken from the tank of the bioreactor container to transfer them to the automatic selection device.
• the automatic dynamic living cell selection device includes in particular (a) at least a first and at least a second culture container intended to receive a culture
• a system of conduits comprising means for connecting, as desired, one of the two culture vessels to the source of medium such as valves as well as the two culture vessels between them and for connecting, as desired, the other culture vessel. culture at the source of the sterilizing agent.
• the gas used can be adapted to aerobic or anaerobic living cells.
• two connecting pipes are provided between the two culture vessels which comprise a common section of pipe.
• an evacuation pipe is provided on the common pipe section through which the cultures can be taken from the culture vessels.
• the living C2 cells improving the reaction are preferably removed by this line.
• the bioreactor device and the automated selection device are supplied with the same substrate, the bioreactor operating continuously, the rate of supply of substrate applied to the line d he arrival of substrate is identical to that applied to the line of racking of culture medium.
• An automated genetic selection device for living cells C2 which can be used is in particular that described in WO-A-00/34433 and which can function according to culture conditions such that the selection device always favors variant living cells called “dynamic” which are better and better adapted to the culture conditions maintained in the bioreactor container.
• all or part of the culture present in the automated selection device is periodically transferred to the bioreactor container.
• the dynamic variant living cells with high growth rate selected by the selection device are periodically inoculated into the bioreactor container where they supplant the static living cells with lower growth rate present in the tank.
• the relationship between the growth rates of living cells present in the tank and living cells with increased growth rate selected by the selection device means that the living cells from the selection device rapidly supplant the living cells present in the tank. bioreactor.
• the growth rate of living cells from the selection device will always be at least equal to the maximum growth rate of living cells present in the bioreactor container.
• the growth rate of living cells from the selection machine is equal to the growth rate of living cells present in the bioreactor container, then all living cells will evolve at the same time ; if it is higher, then the living cells from the selection automaton will take precedence over those already present in the bioreactor container.
• the automated selection device by periodically transferring living cells from the bioreactor container to the automated selection device, it is certain to put the two populations of living cells in competition and to select from the possible variants originating from the bioreactor container and those from the selection device. automated living cells best suited to the conditions of the bioreactor.
• the performance of the bioconversion or biodegradation process conducted in the bioreactor container is therefore at worst maintained but usually continuously improved thanks to the periodic replacement of the active living cells present in the bioreactor container by active living cells originating from the automated selection device, always more efficient because they are always better adapted to the culture conditions. Furthermore, the inoculation being repeated periodically and consequently with living cells which are better and better adapted to the culture conditions, the automated selection device guarantees the preponderance of the most active living cells with respect to the substrate present in the container. bioreactor.
• a small automated selection device for example with culture vessels of only 25 ml, is sufficient to operate effectively a bioreactor container such as the aeration tank of a 100 m 3 wastewater treatment plant . It is possible, of course, to use culture vessels of larger volume, for example 1 liter.
• the living cells C2 used improving the bioconversion reaction can in particular be produced by implementing a method comprising the following steps:
• biomass container designates for example the aeration basin of a treatment plant, the methanization basin of an anaerobic biological treatment unit, a lagoon, a plan of water, a tank for example of 0.5 liters to 100 m 3 , in particular of 1 liter to 100 m 3 , particularly of 5 liters to 50 m 3 and very particularly of 10 liters to 50 m 3 or a fermenter for example from 0.5 liters to 100 m 3 , in particular from 1 liter to 100 m 3 , particularly from 5 liters to 50 m 3 and very particularly from 10 liters to 50 m 3 .
• the term “bioreactor container” designates for example the aeration basin of a treatment plant, the methanization basin of an anaerobic biological treatment unit, a lagoon, a plan of water, a tank for example of 0.5 liters to 100 m 3 , in particular of 1 liter to 100 m 3 , particularly of 5 liters to 50 m 3 and very
• “Substrate” designates a medium containing a compound whose metabolic conversion is envisaged, in particular water of industrial origin such as, for example, washing water from hydrocarbon storage tanks, washing water from production facilities for pharmaceutical intermediates, rinsing water from filter cakes, fume washing water from chemical production, effluents from de-icing of aircraft, water of municipal origin such as domestic waste water, an accidental pollutant from l environment such as the presence at sea of a slick of hydrocarbons or other chemicals originating respectively from the sinking of an oil tanker or a chemical tanker, chemical effluents spilled on the ground following an accident involving a tank transport (road or rail), soil polluted with heavy metals or dioxin.
• substrate also denotes a compound whose metabolic conversion is envisaged, for example glucose used for the production of biomolecules of industrial interest such as lysine, xanthan, alginates, polyols such as glycerol, hygromycin, ethanol used for the production of vinegar by acetic fermentation, oxalic acid used for biohydrometallurgy applications , pectins and carrageenans.
• biomolecules of industrial interest such as lysine, xanthan, alginates, polyols such as glycerol, hygromycin, ethanol used for the production of vinegar by acetic fermentation, oxalic acid used for biohydrometallurgy applications , pectins and carrageenans.
• substrate also designates a medium containing living or dead cells whose metabolic conversion is envisaged, for example an activated sludge from urban or industrial waste water, lignocellulosic derivatives derived from the paper industry, solid or pasty products from the food industry, for example plant biomass, in particular cut grass), grains, yeasts, molasses, or by-products from the fishing industry such as chitinous derivatives, for example those from crab or shrimp shells.
• substrate also designates polluting molecules such as volatile organochlorine compounds (such as chlorinated solvents and CFCs), organochlorine pesticides (such as DDT); halogenated polycyclic aromatic hydrocarbons (such as PCBs, dioxins and furans); solvents (such as benzene, toluene, xylene), organochlorine or organophosphorus phytosanitary compounds.
• volatile organochlorine compounds such as chlorinated solvents and CFCs
• organochlorine pesticides such as DDT
• halogenated polycyclic aromatic hydrocarbons such as PCBs, dioxins and furans
• solvents such as benzene, toluene, xylene
• organochlorine or organophosphorus phytosanitary compounds such as benzene, toluene, xylene
• living cells designates for example one or more bacterial flora such as Sphingomonas wittichii (which catalyzes the bioconversion of dioxin), Pseudomonas putida (which catalyzes the bioconversion of cyanides and cyanates), Agrobacterium radiobacter (which catalyzes the bioconversion of pesticides like bromoxynil), certain strains of Alcanivorax or Acinetobacter (capable of biodegrading many aliphatic hydrocarbons), Xanthomonas campestris (which is involved in the biosynthesis of xanthan) or Sphingomonas paucimobilis (which is involved in the biosynthesis of gellan).
• Sphingomonas wittichii which catalyzes the bioconversion of dioxin
• Pseudomonas putida which catalyzes the bioconversion of cyanides and cyanates
• Agrobacterium radiobacter which
• living cells also refers to animal cells such as mammalian cells (such as HEK-293 cells) for the production of monoclonal antibodies, cellular growth factors of insect cells for the production of recombinant proteins or entomopathogenic viral particles (such as Sf9 cells).
• mammalian cells such as HEK-293 cells
• the term “living cells” also designates plant cells such as Datura plant cells for the production of tropane alkaloids (atropine, hyosciamine and scopolamine), transgenic plant cells for the production of molecules of industrial interest (such as the overproduction of starch by the potato).
• living cells also designates algae such as green algae belonging to the Spirogyra species involved in the biological treatment of effluents containing dyes such as Reactive Yellow 22, cultures of the micro-alga Scenedesmus quadricauda used for the bioconversion of the progesterone, the micro-algae Chlorella vulgaris and Coenochloris pyrenoidosa involved in the biodegradation of p-chlorophenol, the macro-alga Microspora capable of eliminating lead.
• algae such as green algae belonging to the Spirogyra species involved in the biological treatment of effluents containing dyes such as Reactive Yellow 22
• cultures of the micro-alga Scenedesmus quadricauda used for the bioconversion of the progesterone the micro-algae Chlorella vulgaris and Coenochloris pyrenoidosa involved in the biodegradation of p-chlorophenol, the macro-alga Microspora capable of eliminating lead.
• living cells similarly designates yeasts such as Saccharomyces cerevisiae used for the production of bioethanol from glucose or for the production of xylitol from glucose, Candida tropicalis YMEC14 used for the biodegradation of phenol compounds (from production olive oil), Candida famata used for the biodegradation of nitrilated compounds.
• living cells designates fungi such as Penicillium janthinellum capable of producing a xylanase, an enzyme depolymerizing xylan, Streptomyces clavuligerus capable of producing cephalosporin C from glucose as the sole carbon source, or Phanerochaete chrysosporium capable of biodegrading dioxin di- and tetrachlorinated.
• living cells also refers to protozoa such as Euglena mutabilis (acidophilic protozoan) involved in the bioconversion of arsenic.
• living cells also denotes a mixture of all the types of living cells mentioned above.
• Periodic inoculation from the automatic living cell selection device is for example carried out every 48 hours, preferably at least once a week, particularly in the at least once a month and especially after each significant improvement in the growth rate of living C2 cells.
• the continuous, semi-continuous or discontinuous treatment processes of a substrate which is the subject of the present invention have very advantageous qualities. They make it possible in particular to biologically control the functioning of a bioreactor of conventional design by exercising control of the living cells present by elimination of the living cells less adapted to the culture medium as a contaminant having a growth rate lower than that of the living cells present in the bioreactor container for example. It is therefore possible to overcome the constraints of sterility.
• a small selection device for example provided with one-liter bioreactor containers, is sufficient to efficiently operate a container such as a body of water with a volume of 4000 m 3 .
• the invention also makes it possible to improve the efficiency of a culture method of conventional design by increasing the activity of the living cells used in the method without recasting the devices used. It is thus possible to increase the production yields of a molecule of interest and / or the rate of degradation of substrates. These qualities are illustrated below in the experimental part. They justify the use of the processes described above, for example in the biodegradation of recalcitrant compounds.
• the device of the invention makes it possible to maintain living cells in a bioreactor container dedicated to the bioconversion of waste. specifically adapted and efficient vis-à-vis the compounds present in the waste and thus to make possible the bioconversion of waste traditionally destroyed by incineration.
• the invention makes it possible to improve the efficiency of a culture method of conventional design by increasing the activity of the living cells used. in the process without redesigning the implementing devices.
• These qualities also justify the use of the processes described above, for example in improving the operation of biological effluent treatment stations. Indeed, the proper functioning of treatment plants for urban or industrial effluents can be affected by the accidental presence in the effluents of recalcitrant compounds vis-à-vis the living cells present. Provision can be made to remedy this problem by adding a device as described above.
• the bioreactor container is in this case materialized by the existing aeration tank of the treatment plant.
• the automated selection device can be fed by a tapping located upstream of the aeration system in a primary settling tank for example.
• the reciprocal inoculation of the selection device and the bioreactor container is carried out as illustrated below in FIG. 1.
• An external connection line can also be used to supply the automatic device with a substrate modified with respect to the medium taken upstream of the aeration basin.
• This device can be used to enrich the aeration basins with living cells suitable for the biodegradation of any recalcitrant compounds present in the effluents. These qualities also justify the use of the methods described above, for example in improving the performance of biosyntheses.
• the invention can for this purpose be used for improving the performance (yield, growth time) of industrial processes of synthesis by biocatalysis.
• osmotolerant yeasts involved in the production of polyols (sorbitol, mannitol, xylitol, etc.)
• culture times varying from 4 to 5 days are observed on glucose concentrations close to 30 g / l.
• the invention it is possible to bring the yeasts into contact with increasing concentrations of glucose so as to orient their natural metabolism towards the production of the desired extracellular or intracellular metabolite, while increasing their growth rate, that is, by reducing the time required for cultivation.
• This improvement results in an increase in the productivity of the equipment used and in a reduction in the cost price of biosynthesis.
• the method according to the invention can be implemented over long periods and even indefinitely.
• the present application also relates to a device for culturing living cells comprising: A: a selection device preferably comprising two or more receptacles making it possible to receive and maintain cultures of living cells in suspension, - a set of means making it possible to supply these containers separately with cleaning or neutralizing sterilization fluids, a set of means making it possible to supply these containers with gas, a set of means for supplying these containers with substrate, a set of means for transferring the content from one container to the other and vice-versa, - optionally a set of means for evacuating all or part of the content from these containers to another device such as a bioreactor container, a set of means making it possible to evacuate all or part of the content of these containers towards a trash can.
• A a selection device preferably comprising two or more receptacles making it possible to receive and maintain cultures of living cells in suspension, - a set of means making it possible to supply these containers separately with cleaning or neutralizing sterilization fluids, a set of means making it
• - B a bioreactor container
• C a system of means for transferring living cells between the selection device and the bioreactor container
• D optionally a pipe comprising means for connecting the bioreactor container to a solid-liquid separation device such as a settling tank
• E optionally a pipe for discharging the treated fluid (water for example)
• F optionally a temperature regulation device.
• the means for transferring the contents of one container into the other and vice versa can be physical means such as pipes or human means taking samples from one to transfer them to the other.
• the present application more particularly relates to a device for culturing living cells by coupling with an automatic machine for selecting living cells comprising: A: a device for selecting living cells comprising (a) at least a first and at least a second container culture intended to receive a culture (b) a source of gas, (c) a source of medium, (d) a source for a sterilizing agent; and (e) a system of conduits comprising means for connecting, as desired, one of the two culture vessels to the source of medium such as valves as well as the two culture vessels between them and for connecting, as desired, the other culture vessel. culture at the source of the sterilizing agent.
• - B a bioreactor container
• C a system of means for transferring living cells between the selection device and the bioreactor container
• D optionally a pipe comprising means for connecting the bioreactor container to a solid-liquid separation device such as '' a settling tank
• E optionally a pipe for discharging the treated fluid (water for example).
• F optionally a temperature regulation device.
• a draw-off line is installed between the bioreactor container and the automated selection device to allow the removal of living cells present in the bioreactor container in order to make them evolve in the device for automated selection
• an inoculation line is installed between the automated selection device and the bioreactor container to allow the bioreactor container to be sown repeatedly and regularly with living cells having evolved in the automated selection device
• an additional line enriches the automated culture medium with one or more additives
• a tank for collecting rinsing and sterilization effluents collects sterilization and rinsing fluids from the automated selection device
• a set of pumps allows the transfer of different fl uid.
• FIG. 1 represents a schematic view of a device of the invention
• FIG. 2 represents a schematic view of a device for biological purification of wastewater
• FIG. 3 represents a schematic view of an automated selection device described in WO 00/34433.
• FIG. 1 we can observe a bioreactor container 1 connected by a system of return lines 5 and go 6 to an automated crop selection device 2. Pumps 13, 14 are provided on these lines.
• the device can in particular operate as follows: The bioreactor container 1 and the automated selection device 2 are supplied with the same substrate respectively by channels 3 and 4. Since the bioreactor 1 operates continuously, the substrate feed rate applied to line 3 is identical to that applied to line 7 corresponding to the withdrawal of culture medium.
• Line 7 can lead to a solid-liquid separation device, not shown, such as a decanter.
• An inoculation line 5 installed between the automated selection device 2 and the bioreactor container 1 makes it possible to inoculate the bioreactor container 1 repeatedly and regularly with living cells which have evolved in the automated selection device 2.
• a withdrawal line 6 installed between the bioreactor container 1 and the automated selection device 2 makes it possible to take living cells present in the bioreactor container 1 in order to make them evolve in the automated selection device 2.
• An additional line 12 makes it possible to enrich the culture medium of the automaton with one or more additives.
• a bin 8 collects the sterilization and rinsing fluids from the automated selection device.
• a set of pumps 9, 10, 13 and 14 allows the transfer of the various fluids.
• a device for biological purification of waste water can be observed.
• a bioreactor container 1 which is an aeration tank and an automated selection device 2 supplied with the same substrate respectively by channels 3 and 4, inoculation lines and withdrawal 5 and 6 installed between the bioreactor container 1 and the automated selection device 2, and a pipe 15 comprising means for connecting the bioreactor container 1 to a solid-liquid separation device, in this case a decanter 16.
• a pipe 15 comprising means for connecting the bioreactor container 1 to a solid-liquid separation device, in this case a decanter 16.
• FIG 3 shows a first and a second culture container 20, 21, intended to receive a culture 22, a source of gas 23, a source of medium 24, a source 25 for a sterilizing agent, and a system of pipes comprising means for connecting, as desired, one of the two culture vessels 20 or 21 to the source of medium 24 such as valves as well as the two culture vessels 20, 21 between them and for re optionally link the other culture container 20 or 21 to the source 25 of the sterilizing agent.
• the bold lines represent the active pipes during one of the phases of implementation of the process.
• This device allows the provision of a culture 22 in at least a first culture container 20, the continuous supply of the culture 22 in the first culture container 20 with gas from a gas source 23 and regular replenishment of liquids from a source of medium 24, the transfer of culture 22 from the first culture container 20 by connecting lines 28-31 in at least one second culture container 21 by means of a circuit appropriate pipe, connecting the first container of culture 20 with a source 25 for a sterilizing agent, to sterilize the first container the culture 20, the removal of the sterilizing agent from the first culture container 20, the continuous feeding of the culture 22 to the second culture container 21 with gas from the gas source 23 and regular replenishment of liquids from the medium source 24, the return of culture 22 from the second culture container 21 through the connecting lines 28-31 in the first container culture 20 by means of a suitable circuit, the connection of the second culture container 21 with the source 25 for the sterilizing agent, to sterilize the second culture container 21 and the removal of the sterilizing agent from the second container of culture 21.
• Example 1 Bioconversion of wastes from the production of pesticide products
• a 5-liter bioreactor container useful is continuously fed at a fixed flow rate of 0.75 ml / min with a substrate comprising waste from the production of pesticide products.
• the analysis of this waste highlights the following chemical compounds: alcohols (ex: 2-butoxyethanol), alkanes (ex: propane-2,2-dimethoxy), chlorophenols (ex: 2,4-dichlorophenol), aromatics (e.g. 1, 1'-biphenyl, 1-methylnaphthalene, 2-methylnaphthalene, 2-ethylnaphthalene), bromine compounds (e.g. benzonitrile-3,3-dibromo-4-hydroxy) and pesticides (e.g.
• the bioreactor container is inoculated with 10 mL of a mixture of living cells of microorganisms isolated from samples from different ecological niches or activated sludge from treatment plants; these living cells are selected because they are capable of degrading waste.
• the continuous regime is maintained by setting a residual COD of
• an automated selection device of the type described in FIG. 1 of WO-A-00/34433 is also fed with 25 ml culture vessels with the same substrate and inoculated with the same mixture of living cells as above. At the start, in the two containers we therefore have the same living cells. As soon as the turbidity setpoint is reached for the selection system (detection at the turbidostat), the bioreactor container is automatically inoculated with 10 mL of the medium present in the automated selection device.

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• 2003
  • 2003-10-01 FR FR0311502A patent/FR2860510B1/fr not_active Expired - Fee Related
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