WO2016080487A1 - Appareil de réaction biologique, procédé de réaction biologique, structure poreuse soutenant un micro-organisme aérobie ou éventuellement anaérobie utilisée dans l'appareil de réaction biologique et procédé permettant la fabrication de la structure poreuse - Google Patents

Appareil de réaction biologique, procédé de réaction biologique, structure poreuse soutenant un micro-organisme aérobie ou éventuellement anaérobie utilisée dans l'appareil de réaction biologique et procédé permettant la fabrication de la structure poreuse Download PDF

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WO2016080487A1
WO2016080487A1 PCT/JP2015/082548 JP2015082548W WO2016080487A1 WO 2016080487 A1 WO2016080487 A1 WO 2016080487A1 JP 2015082548 W JP2015082548 W JP 2015082548W WO 2016080487 A1 WO2016080487 A1 WO 2016080487A1
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porous structure
biological reaction
microorganism culture
culture solution
micro
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PCT/JP2015/082548
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English (en)
Japanese (ja)
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信秀 国友
宏記 藤井
伸宏 田中
正守 樋口
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三菱化学エンジニアリング株式会社
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Publication of WO2016080487A1 publication Critical patent/WO2016080487A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
    • C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
    • C12N11/082—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer obtained by reactions only involving carbon-to-carbon unsaturated bonds
    • C12N11/084—Polymers containing vinyl alcohol units
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/04—Apparatus for enzymology or microbiology with gas introduction means
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/12—Apparatus for enzymology or microbiology with sterilisation, filtration or dialysis means
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof

Definitions

  • the present invention relates to a biological reaction apparatus and a biological reaction method using the biological reaction apparatus, and a porous medium in which an aerobic or facultative anaerobic microorganism is previously supported in a microorganism culture solution before the biological reaction.
  • the microorganism culture solution contains micro-nano bubbles.
  • aerobic or facultative anaerobic microorganisms are referred to as “aerobic microorganisms, etc.”
  • porous structures that do not carry aerobic or facultative anaerobic microorganisms are simply referred to as “porous structures”.
  • the present invention also relates to a support structure and a method for producing the support structure used in this bioreaction apparatus or bioreaction apparatus, a porous structure, a culture solution containing aerobic microorganisms and micro-nano bubbles, and the like. By contacting the aerobic microorganisms or the like to the depths of the pores of the biological porous structure before use in biological reactions.
  • Biological reactions differ from chemical reactions in that the reaction itself is slow, but because it does not use much energy and many chemical substances, it is a mild and meaningful reaction for the environment.
  • the biological reaction generally has a problem that the reaction is mild and slow. That is, for chemical reactions, a reaction within one hour is often sufficient, whereas in the case of biological reactions, a reaction time of several days or longer, several days or particularly several weeks or longer is required. There is also. For this reason, it is required to perform biological reactions efficiently and economically.
  • Patent Documents 1 to 3 disclose that in the cultivation of microorganisms, the presence of micro-nano bubbles or nano-bubbles in the culture medium promotes the activation of microorganisms and the like. It is disclosed that the efficiency and the reaction time are reduced.
  • Patent Document 1 in a batch method, a culture solution is extracted from a culture tank, filtered with a bacterial cell filter to obtain a filtrate, and micronano bubbles are mixed with the filtrate and returned to the culture tank. It is described to do.
  • Patent Document 2 describes that micro-nano bubbles and nano bubbles are mixed with the culture liquid before supplying the culture liquid to the culture tank.
  • Patent Document 3 supplies the culture liquid to the culture tank. It is described that the micro / nano bubbles are mixed in the previous stage.
  • Patent Documents 4 to 5 that a microorganism is supported by a porous carrier in a biological reaction.
  • Patent Document 4 describes that a microorganism used for wastewater treatment is attached to and supported on a polyvinyl alcohol-containing hydrogel
  • Patent Document 5 describes a citrate-fermenting bacterium.
  • treated water containing saccharides and saccharides is brought into contact with a polyvinyl alcohol-based porous gel so that citric acid-fermenting bacteria are supported on the polyvinyl alcohol-based porous gel.
  • the aerobic microorganism can be transformed into a polyvinyl alcohol-based hydrogel or polyvinyl by simply bringing the bacteria into contact with and supporting the polyvinyl alcohol-based hydrogel or polyvinyl alcohol-based porous gel. It cannot be supported to the back of the pores of the alcoholic porous gel.
  • the subject of the biological reaction apparatus of this invention and the biological reaction method using this biological reaction apparatus is the organism which can perform a biological reaction using an aerobic microorganism etc. efficiently and economically further using a micro nano bubble. It is to provide a reaction apparatus and a biological reaction method.
  • Another object of the porous structure carrying the aerobic microorganisms and the method for producing the porous structure of the present invention is to be used in a biological reaction apparatus or a biological reaction method to efficiently and economically perform a biological reaction.
  • An object of the present invention is to provide a porous structure and a method for producing the porous structure that can be performed.
  • the biological reaction apparatus of the present invention and the biological reaction method using this biological reaction apparatus have a supporting structure in which aerobic microorganisms and the like are supported at a high density in the microorganism culture solution.
  • the microorganism culture solution contains micro / nano bubbles.
  • the porous structure carrying the aerobic microorganisms of the present invention is characterized by carrying the aerobic microorganisms etc. at a high density, and the method for producing the porous structure comprises a porous structure.
  • the structure is brought into contact with a culture solution containing aerobic microorganisms and micro / nano bubbles.
  • micro nano bubble of the present invention means “micro bubble” and / or “nano bubble”. While “normal bubbles” rapidly rise in water and burst and disappear on the surface, microbubbles with a diameter of 50 ⁇ m or less called “microbubbles” shrink in water and disappear. Together with free radicals, “nanobubbles”, which are ultrafine bubbles having a diameter of 100 nm or less, are generated, and these “nanobubbles” remain in water for a certain amount of time.
  • Nano bubbles which are very small bubbles, are also called “ultra fine bubbles”.
  • ISO International Organization for Standardization
  • the creation of an international standard for fine bubble technology is being considered, and once the international standard is created, the name of “nanobubble”, which is currently commonly used, There is a possibility that it will be unified into “Ultra Fine Bubble”.
  • micro / nano bubbles of the present invention can be formed from air or oxygen, but oxygen is preferably used from the viewpoint of efficiently supplying oxygen necessary for respiration to aerobic microorganisms and the like.
  • Patent Documents 1 to 3 utilize these micro-nano bubbles and the action of nano bubbles activating microorganisms.
  • micro-nano bubbles are deep inside the pores of the porous structure.
  • the bioreaction using an aerobic microorganism etc. is performed efficiently and economically using the effect
  • a porous structure having a high loading density in which aerobic microorganisms or the like are loaded in advance into the pores before the biological reaction using micro-nano bubbles is used.
  • the density of aerobic microorganisms etc. in a microorganism culture tank can be made high in a porous structure, the efficiency of a biological reaction can be improved.
  • the filtration step of separating the aerobic microorganisms and the like from the microorganism culture solution and collecting the filtrate can be easily performed. Can be carried out efficiently and economically without causing any significant stress or damage to the body.
  • the microorganism culture solution containing micro-nano bubbles since the microorganism culture solution containing micro-nano bubbles is used, the micro-nano bubbles penetrate deep into the pores of the porous structure, and the aerobic microorganisms supported in the deep pores of the porous structure Since oxygen necessary for respiration can be sufficiently supplied to the living body, the efficiency of the biological reaction can be increased.
  • FIG. 1 It is a photograph which shows the cross section which passes along the center point of the porous structure after carrying aerobic microorganisms etc. by Example 1.
  • FIG. 2 It is a photograph which shows the cross section which passes along the center point of the porous structure after carrying an aerobic microorganism etc. by the comparative example 1.
  • FIG. 1 It is a photograph which shows the cross section which passes along the center point of the porous structure after carrying an aerobic microorganism etc. by the comparative example 1.
  • the main features of the biological reaction apparatus of the present invention and the biological reaction method using this biological reaction apparatus are as described above. 1) The presence of a supporting structure having a high supporting density in which aerobic microorganisms are supported at a high density in the microorganism culture solution; 2) Inclusion of micro / nano bubbles in the microorganism culture solution; However, these items will be described in turn below.
  • Support structure has a high support density in which aerobic microorganisms and the like are supported in advance before being used in biological reactions.
  • the support structure is composed of a porous structure and a support structure. It can be produced by bringing it into contact with an aerobic microorganism or the like and a culture solution containing micro-nano bubbles.
  • porous structure it is preferable to use those having pores, particularly those having a three-dimensional solid network structure having continuous pores.
  • oxygen is not sufficiently supplied to the inside of the pores, so that aerobic microorganisms and the like can only be supported on the surface of the porous structure. .
  • a culture solution containing aerobic microorganisms and micro-nano bubbles as a culture solution to be brought into contact with the porous structure, oxygen can be supplied to the depths of the pores of the porous structure. Aerobic microorganisms and the like can be supported to the depths of the pores, and the density of aerobic microorganisms and the like can be increased.
  • target product a reaction product such as an aerobic microorganism obtained by a biological reaction
  • target product a biological reaction
  • the aerobic microorganism is removed from the microorganism culture solution.
  • a method for supporting aerobic microorganisms or the like in the porous structure a method of adding aerobic microorganisms or the like to the culture solution, culturing and growing the aerobic microorganisms or the like, and then contacting and supporting the porous structure, Alternatively, an aerobic microorganism or the like and a porous structure are added to the culture solution, and the aerobic microorganism or the like is supported while being cultured and grown.
  • the former method is performed by the following procedure. a1) First, sterilize a porous structure, a culture solution, etc. so that microorganisms other than the required aerobic microorganisms, bacteria, etc. do not exist. b1) Next, aerobic microorganisms or the like are cultured and grown while adding aerobic microorganisms or the like to the culture solution and containing the micro / nano bubbles in the culture solution. c1) At the stage where aerobic microorganisms or the like have grown to a certain concentration or higher, the medium is brought into contact with the porous structure while containing micro-nano bubbles.
  • the latter method is performed in the following procedure. a2) First, sterilize the porous structure, the culture solution and the like so that microorganisms other than the required aerobic microorganisms, bacteria and the like are not present. b2) Next, a porous structure is added to the culture solution, and the micronano bubbles are contained in the culture solution. c2) Add aerobic microorganisms or the like to the culture solution, and culture and proliferate the aerobic microorganisms or the like while containing micro-nano bubbles in the culture solution.
  • the culture solution used in the above method mainly contains saccharides and a nitrogen source.
  • saccharides saccharides such as maltose, sucrose, glucose, fructose, and mixtures thereof are usually used.
  • concentration of saccharides in the culture solution is not particularly limited, but is set to 0.1 to 10 w / v%. preferable.
  • nitrogen source ammonium chloride, ammonium sulfate, corn steep liquor, yeast extract, meat extract, peptone or the like is used, and it is preferably set to 0.1 to 10 w / v%.
  • vitamins, inorganic salts, and the like to the culture solution as needed in addition to the saccharides and the nitrogen source.
  • the concentration of the aerobic microorganism or the like added to the culture solution in step b1) and step c2) is not particularly limited, but is preferably 0.5 to 10 g / L, and is preferably 3.0 to 6.0 g / L. More preferably.
  • micronano bubbles are included in the treated water in order to promote the growth of aerobic microorganisms. -The inclusion of micro-nano bubbles can be omitted if the growth can be performed quickly enough.
  • the contact between the culture solution containing the micro-nano bubbles and the porous structure in the above steps c1) and b2) can be performed, for example, by placing the porous structure in the treated water and stirring. .
  • the aerobic microorganisms and the like in the present invention include koji molds (5 to 10 ⁇ m), Bacillus natto (2 to 3 ⁇ m), acetic acid bacteria (5 to 10 ⁇ m) and the like that are conventionally used in technical fields such as brewing and fermentation.
  • koji molds 5 to 10 ⁇ m
  • Bacillus natto (2 to 3 ⁇ m)
  • acetic acid bacteria 5 to 10 ⁇ m
  • various aerobic microorganisms created by gene recombination techniques can be used.
  • the pore size of the continuous pores of the porous structure having a three-dimensional three-dimensional network structure having continuous pores is usually 1 to 30 ⁇ m, preferably 5 to 10 ⁇ m, although it depends on the size of the aerobic microorganisms to be carried. More preferably, it is before and after.
  • Examples of the material of the porous structure suitably used in the present invention include vinyl alcohol resins such as polyvinyl alcohol, ether resins such as polyethylene glycol, acrylic resins such as polymethacrylic acid, acrylamide resins such as polyacrylamide, polyethylene, and polypropylene.
  • vinyl alcohol resins such as polyvinyl alcohol
  • ether resins such as polyethylene glycol
  • acrylic resins such as polymethacrylic acid
  • acrylamide resins such as polyacrylamide, polyethylene, and polypropylene.
  • Olefin resins such as polystyrene, ester resins such as polyethylene terephthalate and polybutylene terephthalate, acrylonitrile resins such as polyacrylonitrile, urethane resins such as polyurethane sponge, calcium alginate, ⁇ (kappa) carrageenan, agar, and cellulose derivatives Sugar, polyester acrylate, epoxy acrylate, urethane acrylate It said photocurable resin, and the like can be exemplified porous inorganic compounds such as activated carbon.
  • a polyvinyl alcohol-based porous gel is preferable in that it has a porous and mesh-like structure up to the inside and a large amount of water can be taken into the gel.
  • the mechanical strength of the porous gel can be improved sufficiently, and it has the strength that it can withstand even with strong agitation during a biological reaction.
  • Formalized polyvinyl alcohol porous gel and acetalization A polyvinyl alcohol-based porous gel is more preferable.
  • Specific examples of the polyvinyl alcohol-based porous gel include, for example, Kuraray Co., Ltd. trade name.
  • the carrying structure of the present invention is one in which the carrying density of aerobic microorganisms and the like is increased by adding micro-nano bubbles to the culture solution.
  • An element for efficiently performing a biological reaction is to increase the density of aerobic microorganisms or the like in a microorganism culture solution. However, if the density of cells is too high, nutrients and Since oxygen is not sufficiently provided, the efficiency of the biological reaction is reduced.
  • the density of cells capable of appropriately culturing aerobic microorganisms and the like is about 3 to 6 g / L.
  • the support structure of the present invention by containing micro-nano bubbles in the culture solution, aerobic microorganisms and the like can be supported deep inside the pores of the porous structure, Depending on the material of the porous structure, etc., the cell density can be about 5 to 6 times the cell density of the culture solution.
  • the shape of the supporting structure is not particularly limited, but for example, a granular shape such as a spherical shape, a rectangular parallelepiped shape or a cubic shape is preferable. If the powder is used, the surface area for fixing aerobic microorganisms and the like can be greatly increased, and the target product can be produced with higher efficiency.
  • the particle size (diameter) of the porous gel powder when dried is preferably 0.5 to 10 mm.
  • the usage structure of the supporting structure may be fixed in the microorganism culture solution with a column, a net or the like, or may be present in a dispersed state in the microorganism culture solution.
  • the support structure obtained in 1) above is housed in a microorganism culture tank together with the culture solution, and the microorganism culture solution contains micro-nano bubbles. Biological reactions take place.
  • a porous structure having a high loading density of aerobic microorganisms or the like in which aerobic microorganisms or the like are previously supported in the biological reaction before the biological reaction is provided.
  • micro-nano bubbles in the microorganism culture solution it is possible to sufficiently supply oxygen necessary for respiration to aerobic microorganisms and the like carried deep inside the pores of the porous structure. Can be improved.
  • a means for containing micro-nano bubbles in a microorganism culture solution a) means for releasing the micro / nano bubbles into the microorganism culture solution in the microorganism culture tank by the micro / nano bubble generator provided outside the microorganism culture tank; b) Means for containing micro / nano bubbles in the culture solution supplied to the microorganism culture tank by the micro / nano bubble generator provided in the pipeline for supplying the culture solution to the microorganism culture tank, c) Means for causing the microorganism culture solution extracted from the microorganism culture vessel to contain micro-nano bubbles by a micro-nano bubble generator and circulating the microorganism culture solution containing the micro-nano bubbles to the microorganism culture vessel d) Pre-containing the micro-nano bubbles A means for supplying the culture solution to the microorganism culture tank through a pipe line can be employed.
  • the means with the least stress and damage given to aerobic microorganisms are the means b) and d) above.
  • stress and damage are given to aerobic microorganisms and the like by the shear force generated by the release of micro-nano bubbles in the means a) and by the shear force generated during filtration in the c) means.
  • this stress and damage can be greatly reduced.
  • micro / nano bubble generating device a known or commercially available device can be used.
  • microbubble generator for example, a sufficient amount of gas is dissolved in water at a certain high pressure, and then the pressure is released to create a supersaturated condition of the dissolved gas.
  • a gas-liquid two-phase flow swirl type micro-bubble generator that utilizes the phenomenon of generating a vortex by generating a water flow, entraining a large bubble in the vortex, and then breaking the vortex into bubbles. Or the like.
  • micro / nano bubble generating apparatus examples include, for example, JP 2007-312690 A, JP 2006-289183 A, JP 2005-245817 A, JP 2007-136255 A, and JP 2009-39600 A. Can be used.
  • the recovery of the target product may be performed batchwise or continuously.
  • the culture tank pump is driven to transfer the filtered filtrate to the filtrate storage tank.
  • the batch type or the continuous type can be selected as appropriate in consideration of the efficiency of biological reaction, the required purity of the target product, economy, and the like.
  • filtration can be performed efficiently and economically, and stress and damage given to aerobic microorganisms can be reduced.
  • a porous membrane made of an organic polymer compound such as polyvinylidene fluoride, or a filtration membrane such as a metal wire mesh can be suitably used.
  • the biological reaction apparatus of the present invention and the biological reaction method using the biological reaction apparatus efficiently and economically perform biological reactions using aerobic microorganisms or the like using micro-nano bubbles. It can be applied not only to the production of foods, chemicals, chemicals and the like using biological reactions such as brewing and fermentation, but also to a biorefinery for producing bioethanol and the like using biomass.
  • porous structure carrying the aerobic microorganisms of the present invention is useful for performing a biological reaction efficiently and economically.
  • FIG. 1 schematically shows a first embodiment of the biological reaction apparatus of the present invention.
  • the first embodiment ⁇
  • the carrying structure 7 is fixed in the microorganism culture tank 1 by the fixing member 8;
  • the micro / nano bubble generating device 3 is provided in a pipeline for supplying the culture solution 6 to the microorganism culture tank 1, and the culture solution 6 supplied to the microorganism culture tank 1 contains micro / nano bubbles.
  • a micro-nano bubble generator 2 is provided outside the microorganism culture tank 1, and the micro-nano bubbles are blown into the microorganism culture solution 5 of the microorganism culture tank 1. It is characterized by.
  • the target product is generated and collected by the following steps. a) When supplying to the microorganism culture tank 1, the micro / nano bubble generator 3 causes the culture solution 6 to contain micro / nano bubbles. b) Even after supplying to the microorganism culture tank 1, the micro / nano bubbles are blown from the micro / nano bubble generator 2 to cause the microorganism culture solution 5 to contain the micro / nano bubbles. c) In the microorganism culture tank 1, a target product is generated by a biological reaction such as aerobic microorganisms supported on the support structure 7.
  • the culture tank pump 10 is driven, the supporting structure 7 detached from the fixing member 8 is separated by the filtration means 9, and the microorganism culture solution 5 containing the target product is collected in the filtrate storage tank 13.
  • the supporting structure 7 having a high supporting density such as aerobic microorganisms and the micro-nano bubbles, in which the aerobic microorganisms are supported in advance before the biological reaction, are provided.
  • the culture solution 5 By using the culture solution 5, a biological reaction can be advanced efficiently and a target object can be obtained efficiently.
  • FIG. 2 schematically shows a second embodiment of the biological reaction apparatus of the present invention.
  • the means for causing the culture solution 6 and the microorganism culture solution 5 to contain micro-nano bubbles in the first embodiment is extracted from the microorganism culture tank 1 by the circulation path 15 and the extracted microorganism culture is performed.
  • the liquid 5 contains micro-nano bubbles, and the microorganism culture solution 5 containing the micro-nano bubbles is changed to means for refluxing the microorganism culture tank 1 (hereinafter referred to as “circulated MNB-containing means”).
  • the microorganism culture solution 5 is extracted through the filtering means 9 and the extracted microorganism culture solution 5 is pressurized and sent to the next step, and the micro-nano bubbles containing the micro-nano bubbles in the pressurized microorganism culture solution 5
  • a generator 4 is provided.
  • the target product is generated and collected by the following steps.
  • a) The culture solution 6 is supplied to the microorganism culture tank 1.
  • the culture tank pump 10 is driven with the valve 11 opened and the valve 12 closed, the carrier structure 7 detached from the fixing member 8 is separated by the filtering means 9, and the microorganism extracted from the microorganism culture tank 1
  • the culture solution 5 is guided to the micro / nano bubble generator 4 to contain the micro / nano bubbles and then refluxed to the microorganism culture tank 1.
  • a target product is generated by a biological reaction such as aerobic microorganisms supported on the support structure 7.
  • Microbe containing target substance by driving culture tank pump 10 with valve 11 closed and valve 12 open, and separating supporting structure 7 detached from fixing member 8 by filtration means 9
  • the culture solution 5 is collected in the filtrate storage tank 13.
  • the circulating MNB containing means by adopting the circulating MNB containing means, 1) Since micro-nano bubbles are contained in the microorganism culture solution 5 from which the support structure 7 is separated, the stress and damage that aerobic microorganisms and the like are subjected to in the filtration step, the micro-nano bubble-containing step, etc. can be reduced. 2) Since aerobic microorganisms are supported on the porous structure, filtration can be performed efficiently and economically. 3) Since the microorganism culture solution 5 is pressurized by the culture tank pump 10 and supplied to the micro / nano bubble generator 4, the generation of micro / nano bubbles can be performed smoothly and economically.
  • the microorganism culture solution 5 in the microorganism culture tank 1 is circulated through the circulation path 15 (that is, extracted from the microorganism culture tank 1 and refluxed), the microorganism culture solution 5 in the microorganism culture tank 1 is agitated. It has the merit of being able to.
  • FIG. 3 schematically shows a third embodiment of the biological reaction apparatus of the present invention.
  • the culture solution 6 supplied to the microorganism culture tank 1 and the microorganism culture tank 1 used in the first embodiment are used.
  • the means for containing the micro-nano bubbles in the microorganism culture solution 5 and the circulating MNB-containing means used in the second embodiment are used in combination.
  • FIG. 4 schematically shows a fourth embodiment of the biological reaction apparatus of the present invention.
  • the support structure 7 is present in a dispersed state in the microorganism culture solution 5, and the microorganism culture vessel 1 further includes a culture vessel agitator 14 for stirring the microorganism culture solution 5. is set up.
  • the support structure 7 is present in a dispersed state in the microorganism culture tank 1 and is stirred by the culture tank agitator 14;
  • the micro / nano bubble generating device 3 is provided in a pipeline for supplying the culture solution 6 to the microorganism culture tank 1, and the culture solution 6 supplied to the microorganism culture tank 1 contains micro / nano bubbles.
  • a micro-nano bubble generator 2 is provided outside the microorganism culture tank 1, and the micro-nano bubbles are blown into the microorganism culture solution 5 of the microorganism culture tank 1. It is characterized by.
  • the object is generated and collected by the following steps. a) When supplying to the microorganism culture tank 1, the micro / nano bubble generator 3 causes the culture solution 6 to contain micro / nano bubbles. b) Even after supplying to the microorganism culture tank 1, the micro / nano bubbles are blown from the micro / nano bubble generator 2 to cause the microorganism culture solution 5 to contain the micro / nano bubbles. c) In the microorganism culture tank 1, a target product is generated by a biological reaction such as an aerobic microorganism supported on the stirred support structure 7.
  • the culture tank pump 10 is driven, the supporting structure 7 is separated by the filtration means 9, and the microorganism culture solution 5 containing the target product is collected in the filtrate storage tank 13.
  • the aerobic microorganism or the like is supported in advance before the biological reaction, and the supporting structure 7 such as the aerobic microorganism or the like and the micro-nano bubbles are contained.
  • the culture solution 5 By using the culture solution 5, a biological reaction can be advanced efficiently and a target object can be obtained efficiently.
  • the support structure 7 is dispersed and stirred in the microorganism culture solution 5 using the culture tank stirrer 14, the aerobic microorganisms or the like supported in the back of the pores of the porous structure are used.
  • the aerobic microorganisms or the like supported in the back of the pores of the porous structure are used.
  • nutrients and oxygen are sufficiently provided and a biological reaction can be promoted.
  • 1) the installation and operation of the culture tank agitator 14 is expensive, and 2) by the agitation of the culture tank agitator 14 Due to the shearing force, aerobic microorganisms and the like are susceptible to stress and damage, and aerobic microorganisms and the like are easily detached from the porous structure.
  • FIG. 5 schematically shows a fifth embodiment of the biological reaction apparatus of the present invention.
  • the means for containing micro-nano bubbles in the culture solution 6 and the microorganism culture solution 5 in the fourth embodiment is changed to circulating MNB-containing means.
  • the target object is generated and collected by the following steps.
  • a) The culture solution 6 is supplied to the microorganism culture tank 1.
  • the culture tank pump 10 is driven with the valve 11 opened and the valve 12 closed, the supporting structure 7 is separated by the filtering means 9, and the microbial culture solution 5 extracted from the microbial culture tank 1 is generated as micro-nano bubbles. After being guided to the apparatus 4 and containing micro-nano bubbles, it is refluxed to the microorganism culture tank 1.
  • a target product is generated by a biological reaction such as an aerobic microorganism supported on the stirred support structure 7.
  • the culture tank pump 10 is driven, the supporting structure 7 is separated by the filtration means 9, and the microorganism culture solution 5 containing the target product is collected in the filtrate storage tank 13.
  • the fifth embodiment has the merit that the biological reaction can be promoted as in the fourth embodiment.
  • the fifth embodiment is provided with a culture tank agitator 14 as in the fourth embodiment.
  • a culture tank agitator 14 as in the fourth embodiment.
  • microorganisms are used. Since the microorganism culture solution 5 in the culture tank 1 can be stirred, stirring using the culture tank stirrer 14 can be reduced.
  • the fifth embodiment uses the circulating MNB-containing means as in the fourth embodiment, it has the advantages 1) to 4) described in the second embodiment.
  • FIG. 6 schematically shows a sixth embodiment of the biological reaction apparatus of the present invention.
  • Means for containing micro-nano bubbles in the culture solution 6 or the microorganism culture solution 5 Means for containing micro-nano bubbles in the culture solution 6 supplied to the microorganism culture tank 1 and the microorganism culture solution 5 of the microorganism culture tank 1 used in the first embodiment, the third embodiment, and the fourth embodiment;
  • the circulating MNB-containing means used in the second embodiment, the third embodiment, and the fifth embodiment is used in combination.
  • the circulating MNB containing means used in the second embodiment, the third embodiment, the fifth embodiment and the sixth embodiment will be further described.
  • the microorganism culture solution 5 in the microorganism culture tank 1 is circulated through the circulation path 15 (that is, extracted from the microorganism culture tank 1 and refluxed), the microorganism culture solution 5 in the microorganism culture tank 1 is agitated. It has the merit that it can be.
  • the circulation path 15 is not limited to a single line, but around the microorganism culture tank 1. It is preferable to provide a plurality of series, and it is more preferable to provide a plurality of series at an equal angle with respect to the central axis of the microorganism culture tank 1. Thereby, the biological culture solution 5 in the biological culture tank 1 is agitated and the dissolved oxygen amount of the biological culture solution 5 is kept uniform without giving excessive stress or damage due to shearing force or the like to the aerobic microorganism. Can do.
  • the microorganism culture solution 5 is extracted from the microorganism culture tank 1 and the microorganism culture solution 5 containing micro-nano bubbles is returned to the microorganism culture tank 1.
  • the suction part and the part where the microorganism culture solution 5 is circulated (hereinafter referred to as “blowing part”), the suction and blowing direction, and the like, are the microorganism culture solution 5 in the microorganism culture tank 1. It can be designed so that the agitation can be performed optimally.
  • a suction part is provided above the microorganism culture tank 1, and the blow part is arranged in the vertical direction in the vicinity of the bottom of the culture tank on the side of the microorganism culture tank 1.
  • an upward flow is generated on the side surface side of the cylindrical microorganism culture tank 1 from the lower blowing part side toward the upper suction part side, and descends toward the center part side of the cylindrical microorganism culture tank 1. A flow can be generated.
  • the vertical and vertical agitation can be uniformly performed over the entire cylindrical microorganism culture tank 1. Well done.
  • the enzyme or the like can be returned to the upper side of the cylindrical microorganism culture tank 1.
  • this circulating MNB-containing means can be used effectively not only for biological reactions, but also for the production of porous structures having a high loading density in the same way as biological reactions.
  • porous structure having a high loading density in which aerobic microorganisms and the like are loaded in advance to the back of the pores before the biological reaction using micro-nano bubbles will be further described.
  • Example 1 Briefly explaining the main points of Example 1 and Comparative Example 1, a case where aerobic microorganisms, a microorganism culture solution, a porous structure and the like are accommodated in a microorganism culture tank and micro / nano bubbles are supplied to the culture tank is implemented.
  • Comparative Example 1 is a case where bubbles having a particle size larger than that of micro-nano bubbles are supplied.
  • a microorganism culture apparatus (microbe culture apparatus BMZ-P manufactured by Able Co., Ltd., internal volume 1000 ml) is used as a microorganism culture tank, and aerobic microorganisms and the like [standard strain of coryneform bacteria (corynebacterium glutamicum)], microorganisms Contains culture solution [synthetic medium containing ammonium sulfate as the main component, glucose concentration: 4%], porous structure [Kuraray Aqua Co., Ltd. “Kragale (registered trademark)”, polyvinyl alcohol-based spherical gel, size diameter of about 4 mm] The liquid volume was 500 ml.
  • this microbial culture tank was reduced to an air flow rate of 250 ml / min and dissolved oxygen concentration below 1 mg / L using a micro / nano bubble generator [nozzle-type micro / nano bubble generator manufactured by OK Engineering, model number: OK-MB 200 ml].
  • the microorganisms were supported on the porous structure for 6 hours by automatically controlling the rotation speed of the culture tank stirrer.
  • coryneform bacteria corynebacterium glutamicum
  • corynebacterium glutamicum which are aerobic microorganisms and the like used in Example 1 and Comparative Example 1
  • the porous structure is cut in a cross section passing through the center point, and the cut surface is By observing the yellow colored state, the state where aerobic microorganisms and the like are supported on the porous structure was evaluated.
  • FIGS. 7 to 9 Porous structure before supporting aerobic microorganisms, porous structure after supporting aerobic microorganisms in Example 1, and porous structure after supporting aerobic microorganisms in Comparative Example 1 Photographs showing a cross section passing through the center point of the body are shown in FIGS. 7 to 9, respectively.
  • the saturation of the cross section of the porous body shown in these photographs was measured.
  • the saturation value of the cross section of the porous structure before supporting the aerobic microorganisms is represented by 1
  • the saturation of the cross section of the porous structure after supporting the aerobic microorganisms in Example 1 is shown.
  • the degree of saturation is about 4, and the saturation of the cross section of the porous structure after supporting aerobic microorganisms or the like in Comparative Example 1 is about 2.
  • the time average value of the rotation speed of the culture vessel agitator automatically controlled so that the dissolved oxygen concentration does not fall below 1 mg / L was 330 rpm in Example 1 and 470 rpm in Comparative Example 1. From this, it is possible to reduce agitation by supporting aerobic microorganisms and the like on the porous structure using micro-nano bubbles as in Example 1, and 1) the operating energy of the culture tank agitator can be reduced. It can be seen that 2) the stress and damage received by the aerobic microorganisms can be reduced, and 3) the aerobic microorganisms and the like are easily carried on the porous structure.
  • a porous structure having a high loading density in which aerobic microorganisms and the like are carried to the depths of the pores is economically, efficiently and stressed by aerobic microorganisms and the like. It can be manufactured with reduced damage.
  • the porous structure is loaded with aerobic microorganisms that are not subject to much stress or damage to the depths of the pores and at a high density, and 2) the micro-nano bubbles are fine in the porous structure.
  • the oxygen necessary for breathing can be sufficiently supplied to aerobic microorganisms and the like carried deep inside the pores of the porous structure, Therefore, the biological reaction can be performed efficiently.
  • aerobic microorganisms and the like are supported on the porous structure, it is possible to reduce stress and damage that aerobic microorganisms and the like are subjected to in a filtration process in a biological reaction, a micro-nano bubble-containing process, and the like.
  • Example 1 and Comparative Example 1 the use of micro-nano bubbles can reduce stirring to keep the dissolved oxygen concentration constant, so that microorganisms can be used using a culture tank stirrer in biological reactions. Even when the culture solution is agitated, 1) the operating energy of the culture vessel agitator can be reduced, 2) the stress and damage to the aerobic microorganisms can be reduced, and 3) aerobicity is released from the porous structure. There are merits such as being difficult.

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Abstract

L'invention porte sur un appareil de réaction biologique et sur un procédé de réaction biologique utilisant l'appareil de réaction biologique, afin d'effectuer de façon efficace et économique une réaction biologique à l'aide d'un micro-organisme aérobie ou éventuellement anaérobie (appelé ci-dessous « micro-organisme aérobie ou similaire »), dans lesquels une structure poreuse ayant une haute densité de charge sur laquelle le micro-organisme aérobie ou similaire est soutenu en profondeur à l'intérieur des pores avant la réaction biologique est placée dans un liquide de culture de micro-organisme et le liquide de culture de micro-organisme est amené à contenir des microbulles/nanobulles. L'invention porte également sur une structure poreuse soutenant un micro-organisme aérobie ou similaire, permettant d'effectuer la réaction biologique de façon efficace et économique, laquelle est amenée à soutenir le micro-organisme aérobie ou similaire à une haute densité de charge avant la réaction biologique. L'invention porte également sur un procédé permettant la fabrication d'une structure poreuse sur laquelle un micro-organisme aérobie ou similaire est soutenu, permettant au micro-organisme aérobie ou similaire d'être soutenu à une haute densité de charge, qui comprend la mise en contact d'une structure poreuse et d'un liquide de culture contenant des microbulles/nanobulles et un micro-organisme aérobie ou similaire.
PCT/JP2015/082548 2014-11-19 2015-11-19 Appareil de réaction biologique, procédé de réaction biologique, structure poreuse soutenant un micro-organisme aérobie ou éventuellement anaérobie utilisée dans l'appareil de réaction biologique et procédé permettant la fabrication de la structure poreuse WO2016080487A1 (fr)

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PCT/JP2014/080574 WO2016079820A1 (fr) 2014-11-19 2014-11-19 Dispositif de réaction biologique, procédé de réaction biologique, structure poreuse porteuse de micro-organisme aérobie à utiliser dans le dispositif de réaction biologique, et procédé de production de structure poreuse

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PCT/JP2015/082548 WO2016080487A1 (fr) 2014-11-19 2015-11-19 Appareil de réaction biologique, procédé de réaction biologique, structure poreuse soutenant un micro-organisme aérobie ou éventuellement anaérobie utilisée dans l'appareil de réaction biologique et procédé permettant la fabrication de la structure poreuse

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US5534143A (en) * 1990-09-18 1996-07-09 Louisiana State University Board Of Supervisors, A Governing Body Of Louisiana State University Agricultural And Mechanical College Microbubble generator for the transfer of oxygen to microbial inocula, and microbubble generator immobilized cell reactor
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