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
  • C12M21/00—Bioreactors or fermenters specially adapted for specific uses
  • C12M21/02—Photobioreactors
• • 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
• • 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

Definitions

• the present invention relates to the field of bioluminescence illumination of a device, in particular a decorative element, a facade of a building or a piece of street furniture.
• Bioluminescence is a chemical reaction governed by a gene that allows certain living organisms to produce light, including marine organisms, algae, jellyfish, squid, fish, or shrimp.
• genes coding for bioluminescence for example, in symbiotic squid bacteria can be inserted into common, non-pathogenic and nontoxic bacteria, encapsulated in a container, with a nutrient solution comprising everything they need to live and produce their light.
• Patent Application US2013045531 describes a lighting assembly comprising a submergible bioreactor and an optical waveguide having a cylindrical structure with a refractive index known to cause the propagation of light longitudinally through a length of the guide. optical wave at an angle of reflection.
• a bioreactor medium having a refractive index different from the known refractive index produces light in the optical waveguide to diffuse the diffusing structure through the inner surface of the optical waveguide and into the medium to another angle of incidence.
• the prior art is known in particular from the international patent application WO2012125953 describing light emitting systems from algae.
• the algae By preparing the algae and applying an appropriate electromagnetic wave, the algae emit light.
• the prepared luminescent algae can also be incorporated into luminescent panels, which can then be integrated in a variety of applications.
• these luminescent panels can be controlled individually and remotely, allowing ease of use and installation.
• these luminescent panels also consume a relatively small amount of energy compared to conventional light sources, creating the potential to save significant amounts of energy and energy costs.
• US patent US5554035 relates to an apparatus for visualizing the luminescence of dinoflagellate algae, incorporated in an aqueous solution contained in a container shaped translucent light bulb.
• bioluminescence activity results in the solutions of the state of the art of a stress which disturbs the survival of the microorganisms, due to the agitation of the biomass observed in the solutions of the prior art.
• the present invention relates to an illumination system of a device, in particular a decorative element, a facade of a building, or a piece of street furniture produced by bioluminescence, characterized in that it includes a bioreactor comprising an arrival of a culture medium and at least one means for sampling a portion of the biomass, as well as connections for connecting a circulation circuit of a medium of culture loaded with bioluminescent microorganisms (that is to say a medium in which the microorganisms evolve), said circulation circuit comprising a set of at least partially transparent conduits and at least one circulation pump and a pump to air, said system comprising a control circuit of the supply of culture medium, the contribution of biomass and removal of biomass and at least one sensor of the biological state of the circulating microorganisms.
• said system comprises other sensors making it possible to intervene in an optimized manner on the biomass culture conditions, such as a biomass oxygenation level sensor, a temperature sensor, a turbidity sensor of the medium. and a biomass
• FIG. 1 shows a schematic view of a system according to the invention.
• the plant comprises a reaction vessel (or bioreactor) (1) containing the biomass and the culture medium and an agitator.
• the reaction vessel (1) receives biomass supplements from a reservoir (2) and liquid culture medium from a reservoir (3).
• the reaction vessel comprises intermittently stirred and oxygenated means, for example with an activation / deactivation time ratio of less than 10% so as to limit the stress of the microorganisms as well as the electrical consumption.
• the removal of excess biomass in the reaction vessel (1) is transferred to an auxiliary container (4).
• the biomass contained in said container (4) can be recovered for the production of biomaterials, energy or heat.
• This container may be equipment for treating biomass, for example by incineration.
• the system may include means for recovering molecules of interest. This recovery step may relate to proteins and / or monoclonal antibodies secreted by the microorganisms, and separated from the biomass by filtration.
• the reaction vessel (1) may have tappings for connecting a circulation circuit (5) of the liquid culture medium containing the biomass.
• the circulation circuit (5) is transparent in the illumination zone.
• a circulation pump (6) provides the flow of biomass in the circulation circuit (5).
• this circuit is formed by a tubular assembly.
• the diameter of such a tubular system can be adapted to the type of device.
• the section may for example be between 10 and 100 millimeters, but may be smaller, for example 5 millimeters or larger, for example 150, 300, 500 millimeters or more.
• the circulation circuit can also take other forms.
• the conduit may have a circular, rectangular, triangular or any other shape of its choice.
• the circulation circuit may consist of two closed plates at their ends between which circulates the solution containing the biomass.
• the term "conduit" means a set of partitions delimiting the passage of the solution containing the biomass regardless of the form (pipe, pipe, passage between two plates ...)
• the circulation circuit (5) is fluid tight and can be gas permeable. It comprises an air pump (13) to ensure the oxygenation of the culture medium.
• the supply of oxygen is indeed essential for the emission of bioluminescence by microorganisms. Oxygen supply may be intermittent or continuous during biomass growth phases but must be continuous during periods during which bioluminescence emission is required.
• said circulation circuit is fluid-tight and gas-permeable, at least partially, to allow CO 2 exchanges with the ambient air.
• the biomass is preferably composed of cyanobacteria, so as to capture the ambient CO 2.
• An electronic circuit also called control system (11) controls the operation of:
• valve (7) controlling the flow rate in the circulation circuit (5)
• valve (8) controlling the addition to the reaction vessel (1) of culture medium from the reservoir (3)
• valve (9) controlling the addition in the reaction vessel (1) of biomass coming from the tank (2)
• valve (10) controlling the evacuation of biomass from the reaction vessel (1)
• valve (14) controlling the air flow.
• the control system (11) may further be configured to automatically control at least one variable of the process.
• the control system (11) may be configured to automatically control at least one parameter among the temperature within the bioreactor, the pressure in the bioreactor, the pH level, the nutrient flow rate, the culture of the medium flow, gas flow, carbon dioxide gas flow, oxygen gas flow, light supply, etc.
• the system can be equipped with sensors to monitor different parameters.
• This control system (11) notably receives signals coming from an optical density sensor (12) arranged on the circulation circuit (5) and / or physicochemical sensors.
• a major advantage of the bioreactor according to the invention lies in the fact that it allows a continuous or semi-continuous culture.
• the good management of the culture in the long term is advantageously ensured thanks to different sensors.
• a culture of bioluminescent bacteria in "batch” mode has no effluent, resulting in an exponential accumulation of bacteria in the reactor until the nutrients are depleted.
• the bacteria then reach a stationary growth phase which is accompanied by metabolic changes preventing the expression of the proteins of interest (in particular bioluminescent proteins).
• metabolic products secreted by bacteria There is also accumulation of metabolic products secreted by bacteria; these products modify the medium and may prevent the bioluminescence reaction.
• different sensors may be associated with the bioreactor of the illumination system according to the invention in order to control the biomass and ensure a good quality of luminescence, among which: a biomass oxygenation level sensor .
• the aeration can be activated or interrupted.
• the interruption of the aeration can, for example, be obtained via a pinch valve which can be provided with a solenoid which makes it possible to cut off the aeration by putting pressure on the air hose.
• a temperature sensor may be an external sensor to monitor the ambient temperature outside the reactor and anticipate its impact on the indoor temperature to maintain an appropriate temperature inside the bioreactor. It is also possible to immerse a temperature sensor in the crop with a special probe.
• a medium turbidity sensor (or optical density as described above). It may be a set of photodiodes consisting of a transmitter and a receiver, both at 940 nm. This type of sensor makes it possible to evaluate the growth of the biomass by using the infrared light coming from the transmitter. The more microorganisms there are, the more the medium becomes murky and less infrared light can pass through the medium to the receptor. With an initial calibration, we can have a relationship between the value obtained with this sensor and the value obtained using a conventional spectrophotometer to calculate the number of bacteria in the medium.
• an irradiance sensor or luminescence sensor. This sensor may be of the light-frequency converter type.
• the sensor emits a frequency that is directly proportional to the intensity of the light that hits the detector. This frequency (Hz) can be translated into irradiance radiometric units ( ⁇ / ⁇ 2 ) using the sensor's technical specifications.
• the sensors can be connected to an interface for displaying the data. These sensors are connected to a control system as described above, which controls the different valves and pumps of the bioreactor in real time.
• the illumination system comprises a set of sensors, namely a biomass oxygenation level sensor, a temperature sensor, a turbidity sensor of the medium and a sensor. irradiance.
• the sensors can be connected to the control system for automated control of the various valves and pumps of the control circuit.
• the temperature sensor should be placed away from heat sources to avoid false reading.
• Infrared LED photodiodes must be placed perfectly in front of each other and the pipes inside the reactor must not block the path between the LEDs.
• the irradiance sensor should be placed in a suitable location where the light produced by the reactor can reach the detector.
• the bioreactor system makes it possible to maintain the bioluminescence for a duration greater than 2 weeks and more generally an unlimited duration.
• the sensors play a major role in this system since they allow optimized management of the biomass by permanently maintaining optimal conditions for the growth of microorganisms and bioluminescence.
• Bioluminescent biomass is a naturally bioluminescent biomass or, preferably, biomass genetically modified to produce bioluminescence, such as:
• photosynthetic organisms algae, microalgae, cyanobacteria
• the culture medium is constituted by way of non-limiting example of a composition comprising:
• the culture media may be freshwater, estuarine or brackish type media or specific marine bacterial species, plankton or algae and / or other microorganisms.
• Culture media may include salts such as sodium chloride and / or magnesium sulfate, macronutrients such as nitrogen and phosphorus-containing compounds, trace elements such as trace metals, e.g. iron and compounds containing molybdenum and / or vitamins such as vitamin B. sub.12.
• Culture media can be modified to accommodate different species and / or to optimize the different characteristics of the crop species, such as growth rate, protein production, lipid production and carbohydrate production. .
• Those skilled in the art will be able to choose a culture medium adapted to the microorganism (s) constituting the biomass.
• the biomass consists of cyanobacteria. These microorganisms have the particularity of capturing C0 2 and emitting 0 2 , and to clean up some dirty water. Thus, in addition to the bioluminescent effect, the device can have a depolluting action.
• the system comprises optically active dopants of the photoluminescent type.
• These dopants are, according to a first variant, constituted by microbeads incorporated in the biomass.
• These materials consist of a polymeric matrix in which are incorporated one or more types of luminescent organic compounds.
• the compounds exhibit an absorption spectrum and an emission spectrum respectively, and the emission spectrum of one of two types of luminescent organic compounds overlaps the spectrum of absorption of the other luminescent organic compound.
• the two spectra corresponding to two curves of intensity of a radiation as a function of the wavelength, have a surface of recovery.
• micro-beads consist for example of nanoparticles of an optically active compound encapsulated in a silicone polymer matrix, ethylene-vinyl acetate (EVA), polyolefins polymethyl methacrylate (PMMA) elastomer, polyurethane, polyamide, ethylene tetrafluoroethylene ( ETFE), polytetrafluoroethylene (PTFE).
• EVA ethylene-vinyl acetate
• PMMA polyolefins polymethyl methacrylate
• ETFE ethylene tetrafluoroethylene
• PTFE polytetrafluoroethylene
• the nanoparticles are, for example, silica nanoparticles, or luminescent organic compounds incorporated inside porous silica, either grafted onto the surface of the nanoparticles.
• the optically active materials are incorporated in a polymer matrix of the biomass circulation conduit.
• conduit and / or the biomass may comprise a diffusing filler, for example silica powder.
• At least one of the faces of the ducts is reflective, for example by depositing a metallized coating; it may be the back side of the ducts.

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• 2016
  • 2016-11-14 FR FR1661006A patent/FR3058867B1/fr active Active
• 2017
  • 2017-11-14 EP EP17804253.7A patent/EP3538637A1/de active Pending
  • 2017-11-14 WO PCT/FR2017/053115 patent/WO2018087504A1/fr active Application Filing

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