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WO2018159577A1 - Composition contenant des microbes destinée au remplissage, et son application - Google Patents

Composition contenant des microbes destinée au remplissage, et son application Download PDF

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WO2018159577A1
WO2018159577A1 PCT/JP2018/007124 JP2018007124W WO2018159577A1 WO 2018159577 A1 WO2018159577 A1 WO 2018159577A1 JP 2018007124 W JP2018007124 W JP 2018007124W WO 2018159577 A1 WO2018159577 A1 WO 2018159577A1
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urea
composition
mineral
filling
present
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PCT/JP2018/007124
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English (en)
Japanese (ja)
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洋平 濱田
辰彦 星野
丈洋 廣瀬
亘 谷川
暁 井尻
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国立研究開発法人海洋研究開発機構
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Publication of WO2018159577A1 publication Critical patent/WO2018159577A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C04—CEMENTS; CONCRETE; ARTIFICIAL STONE; CERAMICS; REFRACTORIES
    • C04B—LIME, MAGNESIA; SLAG; CEMENTS; COMPOSITIONS THEREOF, e.g. MORTARS, CONCRETE OR LIKE BUILDING MATERIALS; ARTIFICIAL STONE; CERAMICS; REFRACTORIES; TREATMENT OF NATURAL STONE
    • C04B28/00—Compositions of mortars, concrete or artificial stone, containing inorganic binders or the reaction product of an inorganic and an organic binder, e.g. polycarboxylate cements
    • C04B28/02—Compositions of mortars, concrete or artificial stone, containing inorganic binders or the reaction product of an inorganic and an organic binder, e.g. polycarboxylate cements containing hydraulic cements other than calcium sulfates
    • C04B28/10—Lime cements or magnesium oxide cements
    • C—CHEMISTRY; METALLURGY
    • C04—CEMENTS; CONCRETE; ARTIFICIAL STONE; CERAMICS; REFRACTORIES
    • C04B—LIME, MAGNESIA; SLAG; CEMENTS; COMPOSITIONS THEREOF, e.g. MORTARS, CONCRETE OR LIKE BUILDING MATERIALS; ARTIFICIAL STONE; CERAMICS; REFRACTORIES; TREATMENT OF NATURAL STONE
    • C04B12/00—Cements not provided for in groups C04B7/00 - C04B11/00
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K17/00—Soil-conditioning materials or soil-stabilising materials
    • C09K17/40—Soil-conditioning materials or soil-stabilising materials containing mixtures of inorganic and organic compounds
    • C09K17/42—Inorganic compounds mixed with organic active ingredients, e.g. accelerators
    • 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
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • 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
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales

Definitions

  • the present invention relates to a filling composition containing microorganisms and a method of using the composition.
  • a high-viscosity filler has been used as a method for filling cracks and gaps in soil and formations.
  • a highly viscous asphalt material is generally used as a pavement on a road surface in view of its flatness and durability.
  • cement-based or lime-based concrete is mainly used as a material for hardening exposed rocks for protecting high slope slope rocks and reducing coastal erosion.
  • a small crack of about several millimeters at most may be filled.
  • Epoxy-based and urethane-based resins are used for filling such cracks.
  • a filler suitable for organic soil and high water content soil a material having light burnt magnesia or phosphate as a main raw material has been developed.
  • Patent Document 1 is a method using resident urea-decomposing bacteria that already exist in minerals. Further, the method described in Patent Document 2 is a method of using a urea degrading bacteria having a predetermined urea hydrolysis rate, Sporosarcina-Pasuturi (Sporosarcina pasteurii) is used as the microorganism.
  • Sporosarcina-Pasuturi Sporosarcina pasteurii
  • urea decomposing bacteria and many known as microorganisms present in the soil, for example, in addition to Sporosarcina microorganism, Bacillus (Bacillus) microorganisms, scan Polo Lactobacillus (Sporolactobacillus) microorganisms, clostripain di
  • Bacillacae such as microorganisms belonging to the genus Clostridium and those belonging to the genus Desulfotomacrum , are capable of hydrolyzing urea into carbonate ions in the presence of water.
  • a wide variety of fillers have been developed for filling and curing the surface of soil and strata.
  • many of them are highly viscous.
  • Such a highly viscous filler is unsuitable for filling objects having gaps or cracks, particularly microcracks such as faults in the formation. That is, a highly viscous filler has a problem that it is difficult to fill a microcrack, and it is difficult to impregnate when press-fitted, making it difficult to perform a wide range of construction.
  • a low-viscosity filler having high penetrating power such as polypropylene glycol diglycyl ester
  • it has a problem in versatility because it is required to be prepared at the time of use. It is not suitable for curing.
  • Patent Document 1 cannot be used for minerals in which resident urea-decomposing bacteria do not exist, and there is a problem that the application target is limited and versatility is poor.
  • the method described in Patent Document 2 uses Sporosarcina pasturi as a urea-degrading bacterium, and according to a study by the present inventors, Sporosarcina pasturi can proliferate under high pressure. There is a problem that the urea resolution is very low. As a result, the method described in Patent Document 2 cannot be used for high-pressure soil or strata, and there is a problem that versatility is poor.
  • the problem to be solved by the present invention is to provide a composition that can be used for filling a gap such as soil or strata, and a method for using the composition, which have a wider range of applications compared to conventional fillers. It is to provide.
  • the present inventors have found that some urea-decomposing microorganisms can grow under high pressure and can decompose urea. Therefore, the present inventors have succeeded in creating a composition that can be used as a filler using the microorganism. Surprisingly, when the simulated fault was filled with the composition, it was found that, in addition to filling the gap in the simulated fault, the strength of the obtained simulated fault was increased over time. . The present invention has been completed based on such successful examples and knowledge.
  • compositions (1) to (3) are provided.
  • the following methods (4) to (7) are provided.
  • a method for filling a mineral comprising a step of filling a gap in or between minerals by bringing the composition of one embodiment of the present invention into contact with the mineral.
  • a method for increasing the strength of a mineral comprising a step of strengthening the mineral by bringing the composition of one embodiment of the present invention into contact with the mineral.
  • a method for producing a mineral reinforcement comprising a step of obtaining a mineral reinforcement by bringing the composition of one embodiment of the present invention into contact with the mineral.
  • composition of one embodiment of the present invention Since the composition of one embodiment of the present invention has a very low viscosity and high penetrability compared to conventional fillers, it is widely used in microcracks and mineral particle gaps in soil and formations. Can be spread and filled.
  • the composition of one embodiment of the present invention can be expected to increase the filling range due to the growth and diffusion of the urea-decomposing microorganisms themselves.
  • each component contained in the composition of one embodiment of the present invention has an advantage that the environmental load is small, and the pH of the composition after filling is about 8, so that the amount of change on the surroundings is small.
  • the method of one embodiment of the present invention it is possible to reinforce the soil or the formation by filling microcracks or gaps in the soil or the formation.
  • the method according to one embodiment of the present invention is applicable to soils and strata under high pressure, for example, the lower and bottom portions of tall stone walls formed by stacking a large number of rocks and stones, deep underground faults, and trench walls.
  • soils and strata under high pressure
  • disaster prevention purposes such as preventing earthquakes and landslides, stabilization of excavation holes, ground improvement, conservation of island coasts, research on fault mechanisms, etc. Can be expected to be promoted.
  • FIG. 1 a shows the electron microscope image (magnification 50) of the rock after immersion in the filling solution as described in the examples.
  • FIG. 1b shows the electron microscope image (magnification 50) of the rock after immersion in a control solution as described in the examples.
  • FIG. 1c shows an electron microscope image (magnification 1000) of the rock after immersion in the filling solution as described in the examples.
  • a portion surrounded by a square in the figure indicates an observation site in FIG.
  • FIG. 1d shows an electron microscope image (magnification 7500) of the rock after immersion in the filling solution as described in the examples.
  • the arrow in the figure indicates a spherical substance estimated to be a urea-decomposing microorganism.
  • FIG. 1 a shows the electron microscope image (magnification 50) of the rock after immersion in the filling solution as described in the examples.
  • FIG. 1b shows the electron microscope image (magnification 50) of the rock after immersion in a control solution as described in the examples.
  • FIG. 2a is a schematic view of a rotary shear friction tester. A portion surrounded by a square in the figure indicates a sample holder portion.
  • FIG. 2b is a schematic view of a sample holder part of a rotary shear friction tester.
  • FIG. 3a is a diagram showing the measurement result of the friction coefficient in the slide-hold-slide test as described in the examples. CM + bacteria in the figure indicates the results when the filling solution is used, and CM_run1 and run2 indicate the results when the control solution is used.
  • FIG. 3b is a diagram showing the measurement result of the maximum friction coefficient in the slide-hold-slide test as described in the example. CM + bacteria in the figure indicates the results when the filling solution is used, and CM_run1 and run2 indicate the results when the control solution is used.
  • composition of one embodiment of the present invention contains at least a urea-decomposing microorganism capable of growing under high pressure, urea, a calcium salt, and components suitable for the growth of the urea-decomposing microorganism.
  • composition of one embodiment of the present invention decomposes urea into carbonate ions in an aqueous solution system by the action of urea-decomposing microorganisms as follows. (NH 2 ) 2 CO + 2H 2 O ⁇ 2NH 4 ⁇ + CO 3 2 ⁇
  • the resulting calcium carbonate exhibits crystalline polymorphism and solidifies to fill the gaps in or with the object to which the composition of one aspect of the invention is applied.
  • Sporosarcina ureae JCM2577 which is a preferred embodiment of a urea-decomposing microorganism that can grow under high pressure, it is well established in minerals, and further, the formation of calcium carbonate is fast.
  • the resulting calcium carbonate has a high hardness and low solubility of calcite, thereby increasing the strength of the solidified object.
  • composition of one embodiment of the present invention is not particularly limited as long as it can be solidified or increased in strength by the formation of calcium carbonate.
  • the geology such as sand, stone, rock, soil, and stratum Examples include inorganic substances generated by the scientific action. In the present specification, these inorganic substances may be collectively referred to as “minerals”. Mineral may refer to a crystalline inorganic substance for academic purposes, but includes an amorphous inorganic substance in the present specification. Further, the mineral is not limited to a naturally occurring one, but includes an artificially manufactured one.
  • the mineral is not particularly limited as long as it is as described above, but is preferably a mineral under high pressure, that is, a pressure higher than atmospheric pressure, more preferably a mineral under a pressure of 0.5 MPa or more, and a pressure of 1 MPa or more. Minerals underneath are more preferred, and minerals under a pressure of 2 MPa or more are even more preferred. A specific example of the mineral is a mineral under a pressure of about 3 MPa.
  • the mineral under high pressure a part or all of the mineral only needs to be under high pressure.
  • the pressure of cracks or gaps in the mineral or between the minerals may be high.
  • minerals under high pressure are high slope slope rocks, exposed rocks to reduce coastal erosion, and the bottom and bottom of stone walls, deep underground faults, trench walls, deep seabeds, drilling holes, etc. But not limited to these.
  • the urea-decomposing microorganism that can be grown under high pressure is not particularly limited as long as it is a microorganism such as an eubacteria, archaea, or fungus having urea decomposing activity that can be maintained or grown under high pressure.
  • the urea degrading microorganisms for example, as eubacteria Sporosarcina microorganism belonging to the genus Bacillus (Bacillus) a microorganism belonging to the genus, vinegar polo Lactobacillus (Sporolactobacillus) a microorganism belonging to the genus, Clostridium (Clostridium) microorganisms of the genus, Death Le photo Mak Lamb (Desulfotomaculum And Bacilliaceae microorganisms such as genus microorganisms, and the like.
  • Bacillus Bacillus
  • Vinpolo Lactobacillus Sorolactobacillus
  • Clostridium (Clostridium) microorganisms of the genus Clostridium (Clostridium) microorganisms of the genus
  • Death Le photo Mak Lamb Desulfotomaculum And Bacilliaceae microorganisms such as genus microorganism
  • the urea-decomposing microorganism that can grow under high pressure is preferably a urea-decomposing microorganism that can be maintained or proliferated under anaerobic conditions and anaerobic conditions from the viewpoint that the object can be a mineral such as a trench or the seafloor, Sporosarcina microorganisms are more preferable, Sporosarcina urea is more preferable, Sporosarcina urea JCM2577 strain is still more preferable.
  • Urea-decomposing microorganisms that can grow under high pressure can be used alone or in combination of two or more.
  • the amount of the urea-decomposing microorganism that can be grown under high pressure in the composition of one embodiment of the present invention is not particularly limited as long as it can form calcium carbonate finally in the aqueous solution system.
  • Urea-decomposing microorganisms that can be grown under high pressure may be those stored at room temperature or low temperature, or may be those in the state of a culture solution in which the stored microorganisms are pre-cultured in advance. A part or all of the preculture solution can be used.
  • the composition according to one embodiment of the present invention is obtained.
  • the content of can be reduced.
  • Sporosarcinina urea JCM2577 strain when Sporosarcinina urea JCM2577 strain is used, the composition of one embodiment of the present invention can be economically improved and the viscosity in an aqueous solution system can be reduced. The advantage is that it can be applied.
  • Urea is not particularly limited as long as it is normally known as having a formula of (NH 2 ) 2 CO, and is in the form of a salt or the like as long as carbonate ions are generated by the action of urea-decomposing microorganisms. May be.
  • the calcium salt is not particularly limited as long as it can generate calcium ions (Ca 2+ ) in an aqueous solution, and examples thereof include calcium inorganic salts such as calcium chloride, calcium hydroxide, calcium nitrate, calcium sulfate, and calcium phosphate. Calcium chloride is preferred from the viewpoint of economy, dissolution rate, and dissolution solution being nearly neutral.
  • the amount of urea and calcium salt used in the composition of one embodiment of the present invention is such that calcium carbonate is finally formed by the action of urea-decomposing microorganisms that can grow under high pressure in an aqueous solution system, and can grow under high pressure.
  • the amount is not particularly limited as long as it does not inhibit the growth of ureolytic microorganisms. For example, it is 100 to 500 mM, preferably 150 to 250 mM, and more preferably 190 to 220 mM.
  • the molar ratio of urea and calcium salt is not particularly limited, and either the molar amount of either one may be large or both may be the same molar amount.
  • Ingredients suitable for the growth of urea-decomposing microorganisms that can be grown under high pressure are particularly limited as long as they are components that promote the growth of urea-decomposing microorganisms, such as those used for culturing urea-decomposing microorganisms as is generally known.
  • components contained in a medium usually used for culturing the urea-decomposing microorganisms such as peptone, polypepton, bacto peptone, fish peptone, animal meat peptone, fish meat extract, animal meat extract, yeast extract, corn steep liquor
  • Animal and plant-derived components such as soybean powder and soybean meal are preferred.
  • the animal and plant derived components can be used alone or in combination of two or more.
  • Ingredients suitable for the growth of urea-degrading microorganisms that can grow under high pressure include, in addition to animal and plant derived components, commonly known microbial medium components such as carbon sources, nitrogen sources, minerals, vitamins, trace amounts A combination of nutrients and the like may also be used. These components may be natural products or synthetic products.
  • the amount of the component suitable for the growth of the urea-decomposing microorganism that can be grown under high pressure is not particularly limited as long as it is an amount usually used for culturing the microorganism for each component.
  • Specific examples of components suitable for the growth of urea-degrading microorganisms that can grow under high pressure include NB medium (Nutrient Broth), but are not limited thereto.
  • the composition according to one aspect of the present invention includes a urea-decomposing microorganism that can grow under high pressure, urea, a calcium salt, and the like, as long as the formation of calcium carbonate from urea and the growth of a urea-decomposing microorganism that can grow under high pressure are not inhibited.
  • other components may be contained.
  • a pH adjuster generally known in the composition of one embodiment of the present invention or A pH buffer or the like may be added.
  • the amount of other components used can be appropriately set by those skilled in the art depending on the purpose of use of the components.
  • composition of one embodiment of the present invention is not particularly limited with respect to its form, and can take the form of, for example, a solid composition or a liquid composition.
  • the composition of one embodiment of the present invention is a solid composition
  • the composition of one embodiment of the present invention is mixed with water or an aqueous solvent at the time of use to obtain an aqueous solution.
  • the solid composition can be prepared by making each component separately, or by making a part or all of them into a solid state and subjecting them to a processing treatment such as a drying treatment if necessary.
  • the liquid composition is preferably prepared as an aqueous solution in which each component of the composition of one embodiment of the present invention is dissolved in water or an aqueous solvent.
  • the state in which the urea-decomposing microorganism capable of growing under high pressure and a component suitable for the growth of the urea-decomposing microorganism are mixed may cause the urea-decomposing microorganism to grow. That is not preferable. Therefore, the composition of one embodiment of the present invention maintains a urea-decomposing microorganism capable of growing under high pressure and a component suitable for the growth of the urea-decomposing microorganism separately, or a mixture of these components. It is preferable to maintain the temperature at such a level that does not grow and die.
  • the composition of one embodiment of the present invention can be a container-packed composition sealed in a container.
  • a container-packed composition By using a container-packed composition, there is an advantage that the composition can be easily stored, transported, and prepared.
  • the container is not particularly limited as long as each component contained in the composition of one embodiment of the present invention is not altered or inactivated.
  • metal such as aluminum, paper, plastic such as PET or PTP, glass, and the like Examples include single-layer or laminated (laminate) film bags, retort pouches, vacuum packs, aluminum containers, plastic containers, bottles, cans, and other packaging containers.
  • a container using a material that blocks passage is preferable.
  • the container to be used is preferably one that has been previously subjected to a conventionally known sterilization treatment such as autoclave or UV.
  • a conventionally known sterilization treatment such as autoclave or UV.
  • the urea-decomposing microorganisms that can be grown under high pressure and the components suitable for the growth of the urea-decomposing microorganisms may be separately packed in two or more containers, and are inserted during use. You may be in the state packed separately in two or more divisions in one container separated by such a partition.
  • composition of one embodiment of the present invention is not particularly limited in its use method.
  • a solid composition it is prepared as an aqueous solution, or in the case of a liquid composition, each component is good.
  • the components contained in the composition of one embodiment of the present invention are individually packaged, they can be used to contact the mineral simultaneously or sequentially.
  • composition of one embodiment of the present invention When the composition of one embodiment of the present invention is brought into contact with minerals, urea is decomposed into ammonium ions and carbonate ions in a metabolic process of a urea-decomposing microorganism that can grow under high pressure, and these ions in water with time. Concentration increases.
  • the composition of one embodiment of the present invention has an advantage that since the viscosity is low, the penetration into the mineral is high and the environmental load is relatively small.
  • the contact between the composition of one embodiment of the present invention and the mineral is caused by carbonation in a portion where the composition of one embodiment of the present invention is in contact with the surface of the mineral, a portion in the mineral, or a crack or gap between the minerals.
  • the contact conditions such as the amount of use, temperature, and time of the composition of one embodiment of the present invention, the contact means, and the like as long as the contact forms calcium.
  • the temperature is preferably 10 to 40 ° C., more preferably about room temperature, and calcium carbonate is stably formed over time.
  • the composition of one embodiment of the present invention in aqueous solution particularly suitable for the growth of calcium salts and ureolytic microorganisms that can grow under high pressure It is preferable to continue press-fitting.
  • composition of one embodiment of the present invention By bringing the composition of one embodiment of the present invention into contact with a mineral, cracks or gaps in the mineral or between the minerals can be filled, and the mineral can be further strengthened. Therefore, specific embodiments of the composition of one embodiment of the present invention are a filling composition and a reinforcing composition, and more specifically, a mineral filling composition and a mineral reinforcing composition.
  • the composition of one embodiment of the present invention can be used as a cement material because it is mixed and bonded together with minerals such as sand and stone and cured.
  • Another embodiment of the present invention is a method and a manufacturing method using the composition of one embodiment of the present invention.
  • Another specific aspect of the present invention is a method for filling minerals, the method comprising filling a gap in or between minerals by bringing the composition of one aspect of the present invention into contact with the mineral.
  • Another specific aspect of the present invention is a method for strengthening a mineral, comprising the step of strengthening the mineral by bringing the composition of one aspect of the present invention into contact with the mineral.
  • the degree of mineral filling and the degree of mineral strengthening are not particularly limited, and slides using an electron microscope observation or a rotary shear friction tester as described in Examples described later.
  • Another specific embodiment of the present invention is a method for producing a mineral fortified product, which comprises a step of obtaining a mineral fortified product by bringing the composition of one embodiment of the present invention into contact with a mineral.
  • Minerals that are 2% or more Examples include reinforcements.
  • Specific embodiments of the production method of the present invention are as follows, for example. Sporosarcina urea JCM2577 strain 1-10mg (wet cell weight), 100-500mM urea, 100-500mM calcium chloride and NB Medium is immersed in a container containing an aqueous solution, or the aqueous solution is cracked Alternatively, the aqueous solution and the mineral are brought into contact with each other by injecting into a portion having a gap and allowing to stand or press-fit at 10 to 40 ° C., preferably room temperature, for several hours to several tens of hours, preferably 48 hours or more.
  • the atmospheric pressure is atmospheric pressure or higher, preferably 1 MPa or higher, more preferably about 3 MPa
  • the Sporosarcinina urea JCM2577 strain can be grown while suppressing the growth of other microorganisms.
  • the formation of calcium carbonate can be confirmed by visual observation or the like by the deposition of calcium carbonate on the surface of the mineral. For minerals in which calcium carbonate deposition has been observed, by confirming the increase in shear friction coefficient and shear maximum friction coefficient by using the rotary shear friction tester described in the examples described later, the deposition of calcium carbonate can be confirmed.
  • the observed mineral can be obtained by evaluating it as a mineral reinforcement.
  • Example 1 Screening for urea-degrading microorganisms
  • Test organism Sporosarcina ureae JCM2577 strain (RIKEN BRC; Bioresource Center, RIKEN Bioresource Center), Sporosarcina pasteurii ATCC11859 strain Lysinibacillus sphaericus) LCM 2502 strain (RIKEN BRC) and Harobachirusu-halophilus (Halobacillus halophilus) LCM20832 shares (RIKEN BRC) was used.
  • microorganisms other than Halobacillus halophyllus used NB medium (Nutrient Broth; Difco; beef extract 0.3 wt%, peptone 0.5 wt%) containing 200 mM urea and 200 mM calcium chloride. .
  • MB medium containing 200 mM urea and 200 mM calcium chloride (Marine Broth; Difco; peptone 0.5 wt%, yeast extract 0.1 wt%, iron citrate 0.01 wt%, sodium chloride 1.945 wt% , Magnesium chloride 0.88 wt%, Sodium sulfate 0.324 wt%, Calcium chloride 0.18 wt%, Potassium chloride 0.055 wt%, Sodium bicarbonate 0.016 wt%, Potassium bromide 0.008 wt%, Strontium chloride 0.0034 wt% %, Boric acid 0.0022 wt%, sodium silicate 0.0004 wt%, sodium fluoride 0.00024 wt%, ammonium nitrate 0.00016 wt%, sodium hydrogen phosphate 0.0008 wt%).
  • Autoclave-sterilized standard rock (Berea sandstone) was immersed in an autoclave-sterilized non-sealed glass bottle containing a medium containing one platinum loop (about 2 mg) of the test microorganism and allowed to stand at room temperature for 12 days. The rock surface after standing was confirmed visually.
  • test microorganisms growth of microorganisms and formation of precipitates could be confirmed.
  • test microorganisms other than Sporosarcina urea JCM2577 were used, the amount thereof was very small. In contrast, the amount of sporosarcina urea urea JCM2577 microbial growth and precipitate formation was significantly greater.
  • Example 2 Evaluation of rock void filling using a filling solution containing urea-decomposing microorganisms.
  • One filling solution Sporosarcina urea JCM2577 strain is pre-cultured aseptically at 30 ° C. in JCM MD22 agar medium (Nutient Agar No. 2; beef extract 10%, peptone 10%, NaCl 5%, agar 15%) did.
  • One platinum loop (about 2 mg) of the cultured Sporosarcina urea was added to an autoclave-sterilized test tube containing 50 ml of NB medium containing 200 mM urea and 200 mM calcium chloride to prepare a filling solution. Further, among the components contained in the filling solution, a solution prepared by removing Sporosarcina urea was used as a control solution.
  • microcrystals of several ⁇ m to several tens of ⁇ m were confirmed on the rock surface when the filling solution containing urea decomposing microorganisms was used.
  • the diameter of the microcrystals was about 10 ⁇ m, and the maximum was about 30 ⁇ m. This is considered to be a microcrystal of calcium carbonate precipitated by the action of urea decomposing microorganisms.
  • the microcrystal of calcium carbonate covered the rock surface and filled the space
  • the imaging result (magnification 1000) of the electron microscope when the filling solution is used is shown in FIG. 1c, and the further enlarged imaging result (magnification 7500) is shown in FIG. 1d.
  • the formed calcium carbonate crystals mainly had a breccia shape, and some of them had a rose shape or a donut shape.
  • a spherical substance having a size of about 1 ⁇ m was observed in the adjacent portion of the breccia-like calcium carbonate crystal (see the arrow in the figure). This is presumed to be a urea-decomposing microorganism.
  • the pH of the solution was 6.20 to 6.34 before standing, and 8.03 after standing. That is, it was confirmed that the solution after standing increased in pH and became weakly basic.
  • the pH of the solution after standing was significantly lower than the pH (12 to 13) when the cementitious material was eluted when a general cementitious material was used. From this, it was shown that the filling solution containing urea-decomposing microorganisms is useful from the viewpoint of environmental load.
  • Example 3 Simulated fault hardening and friction strength evaluation using a filling solution containing urea-decomposing microorganisms] 1.
  • the filling solution of Example 2 and the control solution were used.
  • FIG. 2b shows an outline of the sample holder portion of the testing machine.
  • the sample holder was previously subjected to a sterilization treatment using ethanol.
  • About 15 g of 125 ⁇ m powder sand as a simulated fault was placed on the sample holder, and this was immersed in 4 ml of the filling solution or 4 ml of the control solution.
  • an axial pressure of 3 MPa was applied from the vertical direction of the sample holder, and a slide-hold-slide test (water-containing gouge friction test) was performed.
  • the average displacement speed during sliding was 5 ⁇ m per second, and the hold time was 2, 5, 10, 20, 50, 100, 200, 500, 1,000, 3,600, 10,800 and 43,200 seconds.
  • the sample holder was sealed so that the gas and liquid in the sample holder were not exchanged with the outside.
  • FIGS. 3a and 3b show the results of measuring the friction strength after each hold time.
  • FIG. 3a shows the friction coefficient at the time of sliding
  • FIG. 3b shows the friction coefficient at the time of sliding
  • CM + bacteria in the figure indicates the result when the filling solution is used
  • CM_run1 and run2 indicate the results when the control solution is used.
  • the solution containing urea-decomposing microorganisms, urea, calcium salts and medium components that can grow under high pressure has the effect of filling the gaps of minerals, and surprisingly has the effect of increasing mineral strength. I found it.
  • Compositions and methods according to one aspect of the present invention include minerals under high pressure, such as microcracks and gaps in soils and strata such as the bottom and bottom of tall stone walls, deep underground faults, and trench walls. Because it can be filled and strengthened, it can be used not only for building materials and stones, but also for cementing undersea drilling and disaster prevention.

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Abstract

L'objet de la présente invention est de fournir : une composition qui peut être utilisée dans des espaces de remplissage dans la terre, des formations souterraines, etc., ladite composition ayant une viscosité relativement faible et étant économiquement avantageuse dans une plus large mesure que les matériaux de remplissage précédents, étant peu susceptible de générer des effets secondaires, et présentant en outre une large gamme d'applications; et un procédé d'application de la composition. L'objet susmentionné est atteint grâce : à une composition contenant des microbes décomposant l'urée qui sont aptes à croître sous une pression élevée, de l'urée, un sel de calcium, et des constituants appropriés pour la croissance des microbes décomposant l'urée; et à un procédé de remplissage de minéraux comprenant une étape de remplissage d'espaces dans un minéral par la mise en contact de la composition avec le minéral.
PCT/JP2018/007124 2017-02-28 2018-02-27 Composition contenant des microbes destinée au remplissage, et son application WO2018159577A1 (fr)

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CN114517167A (zh) * 2022-01-20 2022-05-20 吉林省农业科学院 一种秸秆发酵产热方法

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113563015A (zh) * 2021-08-04 2021-10-29 上海天互智慧健康科技发展有限公司 经济环保、高水稳性的工程渣土路面基层的制备方法
CN114517167A (zh) * 2022-01-20 2022-05-20 吉林省农业科学院 一种秸秆发酵产热方法
CN114517167B (zh) * 2022-01-20 2024-02-27 吉林省农业科学院 一种秸秆发酵产热方法

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