WO2004011659A1 - Procede de production microbienne d'hydrogene et appareil de production d'hydrogene - Google Patents

Procede de production microbienne d'hydrogene et appareil de production d'hydrogene

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Publication number
WO2004011659A1
WO2004011659A1 PCT/JP2003/009413 JP0309413W WO2004011659A1 WO 2004011659 A1 WO2004011659 A1 WO 2004011659A1 JP 0309413 W JP0309413 W JP 0309413W WO 2004011659 A1 WO2004011659 A1 WO 2004011659A1
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WIPO (PCT)
Prior art keywords
tank
microalgae
hydrogen
bacteria
carrier
Prior art date
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Ceased
Application number
PCT/JP2003/009413
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English (en)
Japanese (ja)
Inventor
Yoshiharu Miura
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Individual
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Individual
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Publication date
Priority claimed from JP2002217929A external-priority patent/JP2004057045A/ja
Priority claimed from JP2002264655A external-priority patent/JP2004097116A/ja
Application filed by Individual filed Critical Individual
Publication of WO2004011659A1 publication Critical patent/WO2004011659A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • 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
    • C12M21/00—Bioreactors or fermenters specially adapted for specific uses
    • C12M21/04—Bioreactors or fermenters specially adapted for specific uses for producing gas, e.g. biogas
    • 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
    • C12M27/00—Means for mixing, agitating or circulating fluids in the vessel
    • C12M27/18—Flow directing inserts
    • C12M27/24—Draft tube
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P3/00—Preparation of elements or inorganic compounds except carbon dioxide

Definitions

  • the present invention relates to a method and an apparatus for producing hydrogen using microorganisms, and more particularly, to a method for producing hydrogen using microalgae having photosynthetic ability and bacteria having photosynthetic ability. It aims to mass-produce and industrialize. Background art
  • the present invention provides a production method, and further discloses a step of culturing microalgae having photosynthetic ability under light and aerobic conditions to obtain a photosynthetic product in Japanese Patent Application Laid-Open No. 2000-102793. Culturing the microalgae under dark anaerobic conditions to obtain a fermented solution of the photosynthesis; A step of allowing bacteria having photosynthetic ability to act on the fermentation broth under anaerobic conditions.
  • the step of culturing the microalgae under the bright and aerobic conditions and / or the step of causing bacteria to act on the fermentation solution under the bright and anaerobic conditions is carried out in a transparent tower type air-lift tank. Has proposed a production method.
  • Japanese Patent Application Laid-Open No. 10-42881 discloses a hydrogen production system using a marine algae / marine photosynthetic bacterium and the like, in which carbon dioxide is used as a purge gas in a fermentation tank of a green algae fermentation process. What has been proposed.
  • Japanese Patent Application Laid-Open No. 2000-102703 uses a transparent tower-type air lift tank for culturing microalgae under light aerobic conditions and for producing hydrogen by photosynthetic bacteria under light anaerobic conditions. This enables efficient hydrogen production.
  • microalgae cultivation and the production of photosynthetic products such as starch are performed at the same time, so a large amount of light energy is required for cultivation and starch synthesis. Is still not enough. Therefore, there is still room for improvement in terms of mass production of z-elements and industrialization, and further simplification of the production process and reduction of manufacturing equipment are desired.
  • H10-42881 discloses a purge gas. Although carbon dioxide is used to improve carbon dioxide production, green algae are cultivated in a culture tank and starch is accumulated using the cultured green algae fluid, which is necessary for cultivation and starch synthesis. Large amounts of light energy are required, the efficiency of nitrogen production is low, and there is a problem that mass production is not possible. In addition, there is a problem that it cannot respond to mass production and industrialization because it is operated in a batch processing operation.
  • the present invention has been made in view of the above-described problems, and in the production of hydrogen using microalgae, the efficiency of hydrogen production has been improved, and the mass production of hydrogen The task is to plan.
  • the present invention firstly comprises a step of obtaining a hydrogen-producing assimilate by a microalgae having photosynthetic ability, and using the assimilable substance to produce hydrogen by a bacteria having photosynthetic ability.
  • a hydrogen production method comprising a series of steps including a step,
  • the microalgae and / or the cultivation of the bacterium is performed in advance, and the microalgae cells and the bacterial cells are supplied to the above step in a state where the cell concentration is increased by promoting the growth of the cells. It provides a method for producing hydrogen by using microorganisms.
  • the microalgae and bacteria having photosynthetic ability for culturing the cells in advance are microalgae and bacteria in the late logarithmic phase to the stationary phase in which the cell growth has progressed, and have a cell concentration of 1.0 ⁇ 10 8 ce 11. It is present in a saturated state in a solution containing seawater or / and at least a nitrogen supply source, an inorganic component, and a vitamin so that s / ml or more.
  • the above solution preferably contains a small amount of nitrogen source, and the concentration of the nitrogen source such as ammonium chloride is preferably 0.1 mM or less.
  • the salt concentration is set to 1.0% to 10.0%, preferably 3.0% to 5.0%.
  • Salinity refers to the concentration of NaCl.
  • the microalgae can efficiently secrete the seven-source as an extracellular source.
  • microalgae and bacteria in the late logarithmic phase to the stationary phase, the rise time of the reaction in each step can be shortened, and high production efficiency can be obtained from the initial state. Also, when microalgae and bacteria whose cell concentration has been increased by culturing in advance are introduced into the hydrogen production line, the process of culturing microalgae and bacteria in the line is not required, and the 7_K element is produced very efficiently. can do.
  • microalgae and bacteria whose cells have been pre-cultured to increase the cell concentration, light energy is not used for cell growth of microalgae and bacteria in the cell production line, so the final production
  • the production efficiency of hydrogen, which is a product, can be improved, and mass production and industrialization of hydrogen can be realized.
  • the present invention comprises a series of steps including a step of obtaining a hydrogen-producing assimilable substance from a microalga having photosynthetic ability, and a step of producing hydrogen from a photosynthesizing bacterium using the assimilable substance.
  • a hydrogen generation method characterized in that the microalga or the bacterium and the bacterium are fixed to a liquid-insoluble carrier and are present in a solution.
  • the microalgae or bacteria and bacteria immobilized on the carrier are obtained by culturing the cells in advance and increasing the cell concentration.
  • microalgae and bacteria immobilized on the carrier are not limited to microalgae and bacteria that have been cultured in advance to increase the cell concentration, and can be applied to microalgae and bacteria supplied to the above-described series of hydrogen production steps.
  • the microalgae may be used in a carrier of 30 mg dry weight / cm 3 (carrier) to 70 mg dry weight / cm 3 (carrier), preferably 40 mg dry weight / cm 3 (carrier) to 5 O mg dry It is immobilized with a weight of Z cm 3 (carrier).
  • the above bacterium is added to the carrier in an amount of 10 mg dry cell weight / cm 3 (carrier) to 50 mg. It is immobilized at a dry cell weight / cm 3 (carrier), preferably 20 mg dry cell weight / cm 3 (carrier) to 40 mg dry cell weight / cm 3 (carrier).
  • a dry cell weight / cm 3 carrier
  • immobilizing microalgal and bacterial cells on a carrier rather than using them as floating cells, not only stresses the cells but also increases the cell density per unit volume, The activity can be stabilized, and the action of microalgae and bacteria can be maintained for a long time.
  • the immobilized microalgae or bacteria die, they are easily detached from the carrier, and then the microalgae or bacteria are newly grown and adhere to the carrier.
  • the carrier on which the microalgae or bacteria are immobilized can be easily separated from the liquid by solid-liquid separation using a filter or the like. Can be. Therefore, the carrier on which the microalgae or bacteria are immobilized can be collected and recycled, or only the necessary solution can be circulated, or the microalgae and bacteria can be separated so that they do not mix.
  • the above bacteria have an activity of nitrogenase of 200 to 1000 nmo 1 C 2 H 4 / g cell dry wt / hr (this activity is measured by the reduction rate of acetylene to ethylene). It is used in a state of high induction.
  • the nitrogenase may be induced in the bacteria at a high level and then the bacteria may be immobilized on the carrier, or the bacteria may be immobilized on the carrier and the nitrogenase induced in the bacteria at a high level.
  • a porous body is suitably used as a carrier for immobilizing the microalgae and bacteria.
  • the porous body include porous glass beads, polyvinyl alcohol, polyurethane foam, polystyrene foam, polyacrylamide, polyvinyl formal resin L-form, silicon foam, and cellulose L-form.
  • the pore size of the porous body is preferably about 10 to 500 ⁇ m.
  • the shape of the carrier is not limited, but considering the strength and culture efficiency of the carrier, the shape is spherical or cubic. If the size is spherical, the diameter is l mn! 550 mm. In the case of a cubic shape, a square of 1 mm mm50 mm is preferred.
  • the carrier binding method includes a chemical adsorption method or a physical adsorption method in which the resin is adsorbed on an ion-exchangeable resin.
  • Microalgae especially blue algae, are generally sticky and many can be easily adhered and fixed to carriers.
  • the carrier can be easily immobilized by coating the carrier with a positively charged polylysine in advance, and binding the bacteria to the lysine by electrical attraction.
  • ⁇ algae When immobilized by the above method, ⁇ algae can be immobilized at about 5 Omg dry weight / cm 3 (carrier). Bacteria can be immobilized at about 3 O mg dry weight / cm 3 (carrier).
  • the microalgae having photosynthetic ability used in the present invention include: Cyanobacteria and the like are preferred.
  • green algae include Chlamydomonas reinhardtii, Chlamydomonas reinhardtii, Chlamydomonas' moebushii (Chlamydomonas moewusi i), Chlamydomonas moewusi i, Chlamydomonas sp.
  • Chlamydoraonas eugametos Chlamydoraonas eugametos (Chlamydoraonas eugametos) yd Chlamydomo-nas segnis Chlorel la vulgaris belonging to the genus Chlorel la; 1 guus) and Dunaliella laterro-lecta belonging to the genus Dunaliella (Duna-1 iel la).
  • Anabaena variabilis belonging to the genus Anabaena (Anabanena variabi l is) ATCC 29413, Cyanothece sp. ATCC 51142 from the genus Cy-anothece, Synechococcus (Synec- hococcus)
  • the genus includes Synechococcus sp. PCC 7942 belonging to the genus and Anacystis nidulans belonging to the genus Anacystis, and one or more species may be used.
  • Chlamydomonas reinhardi belonging to the genus Chlamydomonas is preferable because of its excellent secretion ability of hydrogen source.
  • the alga Chlamydomonas MG A161 strain and Chlamydomonas W-80 strain are strains isolated from seawater by a conventional method, and have high photosynthetic ability under light and aerobic conditions, as well as dark anaerobic conditions. It is preferable because it has an excellent ability to rapidly adapt to a photosynthetic system when the fermentation under the conditions is fast and when the conditions are returned to the light and aerobic conditions, and it is easy to immobilize on a carrier.
  • Chlamydomonas MG A161 and Chlamydomonas W-80 strains or equivalents are strains that can be easily isolated from seawater samples by a person skilled in the art with photosynthetic ability and resolution.
  • the bacteria having the photosynthetic ability there are used photosynthetic vegetative bacteria and photosynthetic organotrophic bacteria (such as red-colorless sulfur-free bacterium and blue-sliding bacterium), and one or more species may be used.
  • a photosynthetic organic vegetative bacterium is preferably used because a photosynthetic organic substance is used as a substrate (electron donor).
  • a red sulfur-free bacterium belonging to the family Rhodospi-ril laceae is used.
  • green gliding bacteria belonging to the Chloro lexaceae family is used.
  • the photosynthetic organotrophic bacterium can be obtained, for example, by selecting a microorganism that generates hydrogen from a seawater sample using lactic acid (eg, 0.3 mM) as a substrate.
  • lactic acid eg, 0.3 mM
  • photosynthetic bacteria include, for example, Rhodoseudomonas palustris belonging to the genus Rhodoseudomonas and Rhodopseudomonas acidopni la belonging to the genus Rhodoseudomonas.
  • the solution containing seawater and / or a nitrogen supply source, an inorganic component, and a vitamin in which the microalgae and bacteria are present may be a solution containing an appropriate nitrogen supply source, an inorganic component, and a vitamin.
  • Seawater is a medium containing inorganic components.
  • Modified Okamoto medium preferably used is the force ⁇ following composition (hereinafter, referred to as MOM medium): NaC "30 g, C a C 1 2 ⁇ 2H 2 0 ⁇ 2 ⁇ 0 mg, MgSO 4 '7H 2 O ⁇ 250mg , FeSO 4 '7H 2 O ⁇ 20mg, KH 2 P0 4 -40. 8mg, K 2 HP0 4 -495mg, vitamin ... 100 ⁇ g, vitamin ⁇ 12 ⁇ 1 ⁇ g, 1 ⁇ NE C doctor "5m 1, Fine metal mixture A5... 1.0 ml, distilled water... 1000 ml pH 8.0 is also used.
  • composition of the trace metal mixture A5 is as follows.
  • the present invention relates to a microalgae in which cells are cultured in advance and the cell concentration is increased in a state of being fixed to the carrier, or a z-element production method using Z and bacteria.
  • a first step of obtaining a photosynthetic product a second step of using the obtained photosynthetic product by the microalgae to obtain a source of hydrogen generation under anaerobic conditions, and having a photosynthetic ability using the source of photosynthesis. It provides a hydrogen production method that includes three steps: a third step in which hydrogen is produced by bacteria under light and anaerobic conditions.
  • a suspension in which microalgae fixed to the carrier and having a cell concentration increased in advance are present in a solution is supplied.
  • a suspension in which bacteria having a cell concentration increased in advance and fixed to the carrier are present in a solution is supplied. .
  • microalgae and bacteria culture step is not required in a series of hydrogen production steps, and the efficiency is extremely high. Produce hydrogen be able to.
  • microalgae whose cell concentration has been increased are supplied in advance, so that light energy obtained under bright and aerobic conditions is not used for cell growth of microalgae, and generation of photosynthetic products The photosynthetic product can be obtained efficiently. As a result, it is possible to increase the production efficiency of a source of hydrogen production obtained using photosynthesis products.
  • bacteria with an increased cell concentration are supplied in advance, so the light energy obtained under light-anaerobic conditions is not used for bacterial cell growth, but is concentrated on hydrogen generation. Is spent on, and you can get 7 elements efficiently.
  • a first step of obtaining photosynthetic products from the microalgae a second step of obtaining a utilization source by fermentation and decomposition by the microalgae under anaerobic conditions, and a third step of producing hydrogen by bacteria from the purification source Continuous production of three micro-algae with photosynthetic ability used in the first step to the second step, thereby increasing the production speed and increasing the mass production of hydrogen be able to.
  • the above-mentioned assimilable source and microalgae are separated, and the assimilable source and the bacteria having photosynthetic ability that are separated in the third step are allowed to act on the microalgae.
  • the algae is circulated to the first step.
  • the microalgae can be circulated and reused, so that the microalgae having a stable and high cell concentration can always be used.
  • a carbon source is required, so it is preferable to ventilate air or a mixed gas of air and carbon dioxide.
  • the concentration of carbon dioxide in the gas is preferably about 2 to 20%, preferably 2 to; L 0%, and more preferably about 2 to 5%.
  • the cell concentration is determined by preliminarily culturing cells in a state where the cells are fixed to the carrier.
  • Another method of producing hydrogen using enhanced microalgae and / or bacteria is the production of hydrogen by microalgae with photosynthetic ability under light and aerobic conditions, with a liquid phase osmotic pressure of 5.0 atm or more.
  • the present invention provides a hydrogen production method including two steps of producing hydrogen by bacteria having photosynthetic ability using the above-mentioned assimilation source under light and anaerobic conditions.
  • a suspension containing microalgae whose cell concentration has been increased by cell culture in advance in a state of being fixed to the carrier is supplied, and in the second step, the cell is cultured in advance in a state of being fixed to the carrier. It supplies a suspension containing bacteria with increased cell concentration.
  • the method of the present invention omits the second step of obtaining a hydrogen-producing material under anaerobic conditions in the method described above.
  • the omission of the second step is based on the results obtained by the present inventor by repeating experiments.
  • microalgae having photosynthetic ability accumulate photosynthetic products consisting of organic substances such as starch obtained by photosynthesis in cells.
  • photosynthetic products consisting of organic substances such as starch obtained by photosynthesis in cells.
  • increasing the osmotic pressure of the liquid phase, immobilizing the microalgae on the carrier, and increasing the concentration of carbon dioxide in the gas phase may cause stress on the microalgae.
  • the microalgae use light energy to generate a utilization source of hydrogen generation mainly composed of low-molecular-weight organic substances such as glycerol by photosynthesis, and secrete the utilization source out of the cell.
  • a micro-algae with photosynthetic ability can be used as a source of hydrogen production such as glycerol without using the micro-algae under the anaerobic conditions, without going through an enzymatic and decomposition process.
  • (1) the hydrogen production process can be simplified and the 7_K element production efficiency can be improved.
  • the reason why the osmotic pressure is set to 5.0 atm or more is that if the osmotic pressure is lower than this, photosynthetic products generated by the microalgae are accumulated in the microalgae, and the photosynthetic products are used as a source of hydrogen generation. This is because they are not secreted out of the microalgae, and the above-mentioned resources are insufficient, and the hydrogen generation efficiency is reduced.
  • the osmotic pressure of the liquid phase is preferably at least 10.0 atm, more preferably at least 20.0 atm. The higher the osmotic pressure of the liquid phase is, the more effective it is to obtain the assimilation source.
  • the osmotic pressure of the liquid phase is preferably 10.0 atm to 30.0 atm.
  • the volume concentration of carbon dioxide in the first step is 1.0% to 20.0%, preferably 2%.
  • a gas that is between 0% and 15.0% is ventilated.
  • the present invention provides an element production apparatus based on the above-described hydrogen production method.
  • the first of the hydrogen production equipment is
  • the first tank, the second tank, and the third tank are sequentially connected via a pipe or a filter, and the suspension in which the photosynthetic product and the microalga are present is passed from the first tank to the second tank. Then, the suspension from which the microalgae has been removed is circulated from the second tank to the third tank, and a pipe for taking out the hydrogen generated without flowing the bacteria out of the tank is continuously connected to the third tank.
  • a suspension containing the microalgae fixed to a liquid-insoluble carrier and having a cell concentration increased in advance is supplied to the first tank, and the third tank is supplied with a liquid-insoluble carrier.
  • the cell is fixed and the cell concentration is increased in advance.
  • An element production apparatus characterized in that a suspension in which bacteria are present is supplied.
  • the pipe between the second tank and the third tank is provided with a pipe that circulates the separated microalgae to the first tank while interposing the microalgae separating means.
  • the first tank Under aerobic conditions, under the condition that the osmotic pressure of the liquid phase is 5.0 atm or more, the first tank to obtain a source of 7_ nitrogen production by photosynthetic microalgae,
  • a second tank for producing hydrogen by bacteria having photosynthetic ability using the above-mentioned assimilable source under light and anaerobic conditions is provided.
  • the first tank and the second tank are sequentially connected via a pipe or a filter, and
  • the suspension from which the microalgae was removed was circulated from the first tank to the second tank, and a pipe for taking out the generated hydrogen without flowing the bacteria out of the tank was connected to the second tank.
  • the first tank is supplied with a suspension in which the microalgae are fixed in a liquid-insoluble carrier and the cell concentration is increased in advance, and the second tank is fixed on a liquid-insoluble carrier.
  • the present invention also provides a hydrogen production apparatus characterized in that a suspension in which the above-mentioned bacteria whose cell concentration has been increased in advance is supplied.
  • Filters are arranged at the outlets of the first tank and the second tank, respectively, so that the microalgae and the bacteria do not flow out of the tank.
  • a filter made of polysulfone, which is most preferably used, or a filter made of polyethylene, polypropylene, polyvinylidenefluorite or the like is also used.
  • first and third tanks of the above-described hydrogen production apparatus and the first and second tanks of the hydrogen production apparatus to be described later provide a flow for transmitting light energy and heat energy evenly to microalgae and bacteria. Proposed by the applicant It is preferable to use a transparent type air lift tank having a helical flow described in the above-mentioned JP-A No. 2000-110297, but it is also possible to use a stirring type tank.
  • All of the above hydrogen production devices are preferably installed in places where sunlight can be used.
  • light irradiation equipment may be provided as a light energy source when the amount of light is insufficient such as in cloudy weather or at night.
  • Each of the tanks has a discharge port for discharging unnecessary gas.
  • FIG. 1 is a schematic configuration diagram of the hydrogen production process of the first invention.
  • FIG. 2 is a schematic configuration diagram of the hydrogen production process of the second invention.
  • FIG. 3 is a schematic configuration diagram of a transparent tower type airlift tank used in the hydrogen production apparatus of the present invention.
  • FIG. 1 shows a schematic configuration of a hydrogen production device 10 of the first embodiment.
  • the hydrogen production device 10 is a first tank 11 that performs the process of obtaining photosynthetic products under light and aerobic conditions using microalgae with photosynthetic ability.
  • the microalgae with photosynthetic ability using photosynthetic products is used under dark anaerobic conditions.
  • a second tank 12 is provided for performing the step of obtaining a source of hydrogen generation
  • a third tank 13 is provided for performing a step of generating hydrogen under light and anaerobic conditions using bacteria having photosynthetic ability using the source of hydrogen. I have. With these three tanks, the process of obtaining photosynthesis products, the process of obtaining a source of hydrogen generation, and the process of generating z-elements are sequentially performed.
  • the first tank 11 and the second tank 12 are connected by a pipe ⁇ 1, and the second tank 12 and the third tank 13 are connected by pipes ⁇ 2 and ⁇ 3 through a separation device 14. Separation group 14 is also connected to the first tank 11 by pipe # 4.
  • the third tank 13 is provided with a pipe P5 for hydrogen recovery.
  • the temperature of the above tanks 11 to 13 is 15 ° C to 35 ° C, the pressure of the gas phase is about 1 atm, which is normal pressure, and the dilution rate of the suspension is 0.05 hr- 1 to lhr—the average of the suspension.
  • the residence time ranges from 1 ⁇ 11 "to 20111: the pH of the suspension circulating in the first tank 11 is 5 to 8, the pH of the suspension in the second tank 12 is 3 to 6, 3 The pH of the suspension in the tank 13 is set to be 5-8.
  • carbon dioxide as a carbon source air (oxygen), seawater, and microalgae such as green algae having photosynthetic ability are supplied from the inlet of the first tank 11.
  • the microalga to be supplied to the first tank 11 is in a state in which cells are cultured in another container in advance and the cell concentration is increased.
  • the microalgae is fixed in a suspension mainly composed of seawater on a carrier insoluble in the suspension to obtain microalgae in the late logarithmic growth phase or stationary phase in which cell growth has progressed, Cell concentration
  • 1. is a saturated state where the O xl 0 8 cells / ml or more.
  • the first tank 11 is irradiated with light, such as sunlight, so that the conditions are bright and aerobic.
  • the microalgae having the photosynthetic ability described above uses carbon dioxide as a carbon source and uses energy to obtain a photosynthetic product containing starch, and the starch obtained by the photosynthesis is stored in the microalgae.
  • the microalgae are not proliferating, but almost only synthesize starch, and the starch concentration is increased.
  • the starch concentration in the suspension was increased to 1.0 to 3.0 mol 1 Zm1, preferably 1.8 to 2.5 mol Zml, and the second concentration was passed through the pipe P 1 from the first tank 11.
  • the concentration of carbon dioxide is reduced by the amount used for starch synthesis, and the concentration of microalgae is about the same as at the inlet.
  • the second tank 12 no light such as sunlight is irradiated, and no oxygen is present. And 100% carbon dioxide gas is ventilated.
  • Microalgae containing starch which is a photosynthetic product, are degraded under anaerobic conditions, fermented, and ethanol, glycerol, acetic acid, etc. are released from microalgae into suspension.
  • organic acids such as formic acid and lactic acid and alcohols are also released outside the microalgae. Ethanol, glycerol, acetic acid, etc. released in this way are used as sources of hydrogen generation.
  • the organic matter of the utilization source is used as an electron donor for hydrogen production by bacteria having photosynthetic ability.
  • the anaerobic condition in the second tank 12 is a condition under which fermentative production of organic matter is smoothly performed, and particularly preferably a condition under which a large amount of lactic acid is fermented.
  • the anaerobic condition by passing carbon dioxide gas promotes the decomposition of photosynthetic products, while minimizing the generation of hydrogen by microalgae.
  • the time required for fermentation in the second tank 12 may be substantially the same as the time required for starch synthesis in the first tank 11.
  • the product of the concentration of the source at the outlet of the second tank 12 and the flow rate of the suspension is the production rate of the source, and the second tank 12 is the production rate of this source. Is the highest condition.
  • the suspension containing assimilable resources and microalgae flowing out of the second tank 12 is sent to the separation device 14 through the pipe P2, and is fixed to the carrier by the filter 1 provided in the separation device 14.
  • the resulting microalgae are cut off and separated from the suspension containing assimilable resources.
  • the separated microalgae is circulated through the pipe P4 to the first tank 11 and reused in the step of obtaining photosynthetic products.
  • the suspension containing the separated resources flows into the third tank 13 through the pipe P3.
  • the source of inflow from the second tank 12 is reacted with bacteria having photosynthetic ability to generate ⁇ -element.
  • bacteria having photosynthetic ability are immobilized on a carrier insoluble in the suspension, and cells are cultured in advance to increase the cell concentration.
  • the above bacteria are bacteria in the late logarithmic growth phase or stationary phase in which cell growth has progressed, and are present in a suspension mainly composed of seawater, and have a cell concentration of 1.0 X 1 It is saturated so that it is not less than 0 8 cells / ml.
  • This ditrogenase is activated by an activity of 200 to 100 nmol C 2 H 4 / mg cell dry wet / hr (this activity is measured by the reduction rate of acetylene to ethylene).
  • the third tank 13 is irradiated with light, such as sunlight, and is in a light-anaerobic condition.
  • the above bacteria produce hydrogen from light sources such as ethanol using light energy, and the obtained hydrogen is recovered through pipes. Note that a part of the generated hydrogen may be circulated to the third tank 13.
  • microalgae and bacteria having photosynthetic ability are supplied in a state where the cell concentration is increased in advance, so that products required for hydrogen production can be efficiently obtained.
  • the process of obtaining photosynthetic products in the first tank 11, the step of obtaining a source of hydrogen generation in the second tank 12, and the step of generating hydrogen in the third tank 13 are being continuous. Therefore, hydrogen can be produced very efficiently in the end, and mass production of zK element and industrialization of hydrogen production can be realized.
  • FIG. 2 shows a schematic configuration of a hydrogen production apparatus 20 of the second embodiment.
  • the hydrogen production apparatus 20 includes a first tank 21 for generating a source of hydrogen generation using microalgae having photosynthetic ability, and hydrogen generating by bacteria having a photosynthetic ability using the above-mentioned sources.
  • a second tank 22 is provided.
  • the first tank 21 and the second tank 22 are configured to be able to transmit light energy and heat energy evenly to the microalgae and bacteria.
  • a supply pipe 23 for supplying seawater is connected to the first tank 21 and the first tank 21 is connected to the first tank 21.
  • the outlet of the tank 21 is connected to the second tank 22 via the first filter F 1 and the first pipe P ⁇ .
  • the microalgae in the first tank 21 is prevented from flowing out of the tank by the first filter F 1, and only the suspension in the first tank 21 flows out to the second tank 22.
  • the osmotic pressure of the liquid phase is set to 5. 0 atm or more and 30.0 atm or less, and in the present embodiment, 24.6 atm (atmospheric pressure). Further, a gas having a volume concentration of carbon dioxide of 1.0% to 20.0% (5.0% in the present embodiment) is ventilated.
  • the second tank 22 is connected to a second pipe P 2 ′ for hydrogen recovery, and is provided with a discharge pipe 24 for discharging seawater.
  • a second filter F2 is provided at the outlet from the second tank 22 to the second pipe P2 ', and a third filter F3 is provided at the outlet of the discharge pipe 24 from the second tank 22. Is not allowed to flow out of the second tank 22 out of the tank.
  • the temperatures of the tanks 21 and 22 and the residence time of the suspension were the same as in the first embodiment, and the pH of the suspensions in the first tower 21 and the second tank 22 was 5 to 8. It is set to be.
  • carbon dioxide as a carbon source air (oxygen), seawater, microalgae such as algae, etc.
  • air oxygen
  • seawater water
  • microalgae such as algae, etc.
  • the volume concentration of carbon dioxide in the gas to be ventilated is 5.0.
  • the microalgae is the same as in the first embodiment, is present in a suspension in a state of being fixed to a carrier, and has a previously increased cell concentration.
  • the microalgae uses carbon dioxide as a carbon source, generates starch of a photosynthetic product using photoenergy, accumulates in a cell, Glycerite is secreted from microalgal cells into the suspension. Secreted glycerol, etc., is a source of hydrogen production.
  • the suspension containing the microalgae in the first tank 21 and a source of hydrogen production secreted from the microalgae is separated by the filter F1, and the microalgae remains in the first tank 21.
  • the suspension containing the utilization source flows through the first pipe P ⁇ and continuously flows out to the second tank 22.
  • the microalgae since the microalgae are fixed to the carrier, the microalgae can be surely separated by the filter F1.
  • the product of the concentration of the source of hydrogen generation at the outlet of the first tank 21 and the outflow rate of the suspension is the production rate of the source of hydrogen generation, and the first tank 21
  • the conditions are such that the speed is highest.
  • the source of hydrogen generation flowing from the first tank 21 and the bacteria having the photosynthetic ability are allowed to act to generate zK element.
  • the bacteria in the second tank 22 are the same as the bacteria supplied to the third tank of the first embodiment, are present in a suspension in a state of being fixed to a carrier, and have a predetermined cell concentration. It is said that the condition has been enhanced and that ditrogenase has been highly induced.
  • the second tank 22 is in a light-anaerobic condition in which there is no oxygen present due to irradiation with light such as sunlight.
  • the bacteria produce hydrogen using light energy from a source of hydrogen generation such as glycerol and ethanol. ing.
  • the obtained hydrogen is recovered through the second pipe 'P 2' and continuously produces hydrogen.
  • the suspension containing the bacteria is separated by the filter F3, and the bacteria remain in the second tank 22.Seawater, which is the main component of the suspension, is discharged from the discharge pipe 24. I have. A part of the generated hydrogen may be circulated to the second tank 22.
  • the microalgae since the osmotic pressure of the liquid phase of the microalgae having photosynthetic ability is 24.6 atm under bright and aerobic conditions, the microalgae obtains a photosynthetic product by photosynthesis, and under dark anaerobic conditions.
  • a source of hydrogen generation such as glycerol can be obtained by photosynthesis without going through the fermentation process below.
  • the fermentation step performed under dark anaerobic conditions of the first embodiment can be omitted, and a product required for hydrogen production can be efficiently obtained with a simple system.
  • the microalgae and the bacteria are not fixed to the carrier, but are preliminarily cultured to increase the cell concentration. Then, as in the second embodiment, the suspension of the microalgae with the increased cell concentration is increased. Suspended liquid is supplied to the first tank, and the bacteria whose cell concentration has been increased are supplied to the second tank.
  • the cell concentration of the microalgae and bacteria is defined as microalgae and bacteria in the late logarithmic phase to the stationary phase in which the growth of cells has progressed, and the cell concentration is not less than 1.0 ⁇ 10 8 eel Is / ml.
  • the concentration of a nitrogen source such as ammonium chloride is set to 0.1 mM or less.
  • the salt concentration is set to 1.0% to 10.0%.
  • the suspension of the microalgae which is not fixed to the carrier but has a previously increased cell concentration, is similarly used.
  • the solution may be supplied to the first tank, and the bacteria with an increased cell concentration may be supplied to the third tank.
  • the microalgae and the bacterium having photosynthetic ability are not previously raised in cell concentration, but are present in a suspension in a state of being fixed to a carrier insoluble in a suspension.
  • the hydrogen is supplied to the first tank and the second tank of the same hydrogen production apparatus as in the second embodiment. Note that the first embodiment of the hydrogen production apparatus of the first embodiment It may be supplied to the tank and the third tank.
  • the microalgae is 30 mg dry weight / cm 3 (carrier) to 70 mg dry weight / cm 3 (carrier) in the carrier, and the above bacteria is 10 mg dry cell weight / cm 3 (carrier) in the carrier. 5 O mg dry cell weight Z cm 3 (carrier), fixed in this embodiment, ⁇ ⁇ algae as microalgae, fixed at 50 mg dry weight / cm 3 (carrier), bacteria Fixed at mg dry weight / cm 3 (carrier).
  • the carrier is made of a porous polymer material consisting of polynivir formal and has a diameter of 1 mn! It is a sphere of up to 5 O mm or a cube of 1 mm to 5 O mm square. To 500 m. A microalgae having adhesiveness is used as the microalgae for this carrier, and the microalgae is fixed by a carrier binding method.
  • the above-mentioned bacterium has a negatively charged cell surface.
  • the carrier is coated with a positively charged polylysine in advance, and the bacterium is bound to the lysine by electrical attraction and fixed.
  • the nitrogenase is activated by the activity of 500 to 1000 nmo 1 C 2 H 4 / mg cell dry wtZhr and is highly induced. Used. At this time, it is good to induce nitrogenase in the above bacteria at a high level and then immobilize the bacteria on the carrier, but it is also possible to immobilize the bacteria on the carrier and then induce a high level of ditrogenase in the bacteria. .
  • the cell concentration of the microalgae and bacterial cells has not been increased in advance, the cells are not used as floating cells but are immobilized on a carrier to not only stress the cells, but also to increase the cell density per unit volume.
  • the cell activity can be stabilized, and the action of microalgae and bacteria can be maintained for a long period of time.
  • immobilized microalgae When the microbes and bacteria die, they are easily detached from the carrier, and then the microalgae and bacteria are newly grown and adhere to the carrier, so that the action of the microalgae and bacteria can be continuously obtained.
  • the carrier is insoluble in the liquid in which microalgae and bacteria are present, the outflow to the outside of the tank can be blocked by a filter provided at the outlet of the supplied tank.
  • the microalgae can be circulated to the first tank without flowing into the third tank by a separation device interposed between the second tank and the third tank.
  • the first tank 11 to the third tank 13 of the first embodiment, the first tank 21 and the second tank 22 of the second to fourth embodiments are each a transparent tower type airlift tank 30 shown in FIG. Power
  • the air lift tank 30 is installed so as to be irradiated with sunlight in the tank under the bright aerobic condition and the bright and anaerobic condition, and is shielded from the sunlight in the tank under the dark anaerobic condition.
  • each of the above-mentioned tanks is not limited as long as the structure can perform the required bright and aerobic conditions, bright and anaerobic conditions, or ⁇ anaerobic conditions.
  • the transparent tower type airlift tank 30 has a double cylindrical structure in which an inner cylinder 32 is arranged below the outer cylinder 31 with a space enough for liquid to flow.
  • the space between the outer cylinder 31 and the inner cylinder 32 and the inside of the inner cylinder 32 are used as circulation channels for the suspension 33, respectively.
  • a ventilation device 34 is arranged in the lower part inside the inner cylinder 32 to ventilate, and a suspension 33 flows from the lower part to the upper part inside the inner cylinder 32 to generate a flow of the suspension 33.
  • the suspension 33 is circulated downward from the upper surface inside 2 to the flow path between the inner cylinder 32 and the outer cylinder 31.
  • the suspension 33 is configured to be irradiated with the strongest light when flowing through the outer peripheral flow path between the outer cylinder 31 and the inner cylinder 32.
  • a helical mouth promoter 36 for generating a spiral or radial flow is arranged.
  • the transparent tower type airlift tank 30 having the above-mentioned configuration is designed to uniformly apply heat energy to tanks under anaerobic conditions, heat energy and light energy to cells under bright and aerobic conditions and light and anaerobic conditions. Because of the provision of a flow path that can be provided, a suspension containing microalgae having photosynthetic ability circulating in the flow path and a suspension containing bacteria having photosynthetic ability can uniformly and cyclically emit light energy. As a result, photosynthetic ability is enhanced. In addition, the irradiation time of light on microalgae and bacteria can be made longer and the amount of photosynthesis can be increased because microalgae and bacteria are circulated more efficiently than in the case of using a conventional stirred-type culture tank.
  • Example 1 and Comparative Example 1 according to the hydrogen production method of the first embodiment will be described in detail.
  • an air-lift bioreactor with a helical flow promoter with an actual volume of 50 L was used.
  • the height of the transparent tower type air lift tank made of acryl resin was 1.8 m
  • the outer diameter was 0.2 m
  • the diameter of the inner cylinder made of acrylic resin was 0.125 m.
  • a gas inlet which is a ventilation device, was installed at a position of 0.05 m from the bottom of the tank. Specifically, 50 holes with an inner diameter of 0.001 m were drilled in a 0.01 m inner diameter circular tube (circle diameter: 0.1 m), and gas was blown from there. Examples and Comparative Examples are the same.
  • the bright and aerobic conditions were performed using sunlight and a light source (18 W / m 2 ).
  • the main component of the suspension was seawater.
  • the alga Chlamydomonas W-80 strain was used as the microalgae having photosynthetic ability.
  • the bacteria having photosynthetic ability used were oral Dobram sulphide dophyllum W-1S.
  • a porous polymer material made of polyvinyl formal was used as the carrier.
  • Example 1 As shown in Table 1 below, microalgae and bacteria were used in a state where the cell concentration was increased in advance. Using stationary phase microalgae and bacteria, they were immobilized on a carrier.
  • microalgae and bacteria were used in a state where the cell concentration was not high and cells could grow.
  • Microalgae and bacteria in logarithmic growth phase were used and used as floating cells without immobilization on a carrier.
  • Hydrogen was produced by the methods of the above Examples and Comparative Examples, and the rate of hydrogen production and the rate of hydrogen production were measured.
  • the dilution ratio was 0.1 lhr- 1 .
  • Table 2 shows the measurement results.
  • Example 1 had a much higher hydrogen production rate than Comparative Example 1 and could efficiently produce hydrogen. Therefore,
  • 7_ nitrogen can be mass-produced, and it has been confirmed that the method is very useful for industrialization of element production by microorganisms.
  • Example 2 and Comparative Example 2 of the hydrogen production method using the hydrogen production apparatus of the second embodiment will be described in detail.
  • Hydrogen was produced under the conditions shown in Table 3 below, and the hydrogen production rate was measured. Cells used
  • the following experiment was conducted on the hydrogen production method of the second embodiment.
  • an airlift bioreactor with a helical mouth and one promoter having an actual volume of 50 L was used as a first tank and a second tank.
  • the height of the transparent tower type air lift tank made of acryl resin was 1.8 m
  • the outer diameter was 0.2 m
  • the diameter of the inner cylinder made of acrylic resin was 0.125 m.
  • the gas inlet which is a ventilation device, was installed at a position of 0.05 m from the bottom of the tank.
  • 50 holes with an inner diameter of 0.001 m were made in a circular tube with an inner diameter of 0.0 lm (diameter of a circle of 0.1 lm), and gas was blown from there.
  • the bright and aerobic conditions were performed using sunlight and a light source (18 W / m 2 ).
  • the main component of the suspension was seawater.
  • a green alga Chlamydomonas W-80 was used as a microalga having photosynthetic ability.
  • Bacteria having photosynthetic ability were oral doplum sulphide dophyllum W-1S.
  • the osmotic pressure of the liquid phase in the first column was 1.9 atm. Others were the same as in Example 2.
  • Example 2 since the osmotic pressure of the liquid phase in the first column was high, the hydrogen production rate was much higher than that in Comparative Example 2, and the z-element could be produced efficiently. Was confirmed. Therefore, according to the method of the second invention, hydrogen can be mass-produced by a simple system in which the number of columns is small and the number of fermentation steps is reduced, which is very useful for industrialization of hydrogen production by microorganisms. Was confirmed.
  • the present invention uses microalgae and bacteria in a state in which cells have been cultured in advance and the cell concentration has been increased.
  • the light energy obtained below is concentrated on the production of photosynthetic products and the production of hydrogen, so that hydrogen can be obtained efficiently and mass production of hydrogen can be achieved.
  • the cell density per unit volume can be increased, and the cell activity can be stabilized. It has various advantages such that the action of bacteria can be maintained for a long time.
  • microalgae with photosynthetic ability when used under conditions where the osmotic pressure of the liquid phase is high, the microalgae can use light energy to utilize hydrogen as a main source of low-molecular-weight organic substances such as glycerol. Since photosynthesis is performed and secreted outside the cells, the fermentation process can be reduced.
  • carbon dioxide can be recycled and used in flue gas such as thermal power plants.
  • Carbon dioxide etc. can be used as a carbon source, which is very friendly to the global environment.
  • hydrogen can be generated using seawater, and water can be finally obtained by using the generated hydrogen as an energy source, so it can be applied to desalination of seawater.
  • the apparatus for producing hepatocellular microorganisms of the present invention can produce hydrogen very easily and efficiently using microorganisms, and can reduce unnecessary production facilities and the like, thereby greatly reducing production costs. You can also.

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Abstract

L'invention concerne un procédé de production d'hydrogène comportant une série d'étapes consistant à obtenir une source métabolique de microalgue photosynthétique et à faire produire de l'hydrogène par une bactérie photosynthétique au moyen de la source métabolique. Dans ce procédé, la microalgue et/ou la bactérie sont fixées sur un support insoluble dans un liquide et mises en préculture dans ce liquide de façon à élever la densité cellulaire avant de procéder aux étapes décrites ci-dessus. La densité cellulaire à laquelle la microalgue et la bactérie sont amenées correspond à la fin de la phase de croissance logarithmique ou à la phase stationnaire, la croissance des cellules étant réalisée, et se trouvant à l'état saturé, dans de l'eau de mer et/ou dans une solution contenant une source d'azote, des matières inorganiques et des vitamines.
PCT/JP2003/009413 2002-07-26 2003-07-24 Procede de production microbienne d'hydrogene et appareil de production d'hydrogene Ceased WO2004011659A1 (fr)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010030658A3 (fr) * 2008-09-09 2010-07-01 Battelle Memorial Institute Production de matériaux biosourcés au moyen de photobioréacteurs avec des cultures binaires
US8017377B1 (en) * 2008-04-11 2011-09-13 Agoil International, Llc Mass culture of microalgae for lipid production
CN109929897A (zh) * 2019-03-13 2019-06-25 河南农业大学 一种促进hau-m1光合细菌菌群产氢的培养基及其应用
CN109929898A (zh) * 2019-03-28 2019-06-25 河南农业大学 一种促进hau-m1光合菌群节能高效产氢的方法

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EP0077014A2 (fr) * 1981-10-08 1983-04-20 Yoshiharu Miura Procédé de production de l'hydrogène à partir d'algues dans un cycle alternant de clair et obscur
JPH1042881A (ja) * 1996-07-30 1998-02-17 Tadashi Mizoguchi 微生物を用いた有用ガス生産プロセス
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YOKOI H. ET AL.: "Simultaneous production of hydrogen and bioflocculant by enterobacter sp. BY-29", WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY, vol. 17, no. 6, 2001, pages 609 - 613, XP002973396 *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8017377B1 (en) * 2008-04-11 2011-09-13 Agoil International, Llc Mass culture of microalgae for lipid production
WO2010030658A3 (fr) * 2008-09-09 2010-07-01 Battelle Memorial Institute Production de matériaux biosourcés au moyen de photobioréacteurs avec des cultures binaires
CN109929897A (zh) * 2019-03-13 2019-06-25 河南农业大学 一种促进hau-m1光合细菌菌群产氢的培养基及其应用
CN109929897B (zh) * 2019-03-13 2022-08-30 河南农业大学 一种促进hau-m1光合细菌菌群产氢的培养基及其应用
CN109929898A (zh) * 2019-03-28 2019-06-25 河南农业大学 一种促进hau-m1光合菌群节能高效产氢的方法
CN109929898B (zh) * 2019-03-28 2022-08-30 河南农业大学 一种促进hau-m1光合菌群产氢的方法

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