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WO1993013213A1 - Procede de fermentation - Google Patents

Procede de fermentation Download PDF

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Publication number
WO1993013213A1
WO1993013213A1 PCT/JP1987/000156 JP8700156W WO9313213A1 WO 1993013213 A1 WO1993013213 A1 WO 1993013213A1 JP 8700156 W JP8700156 W JP 8700156W WO 9313213 A1 WO9313213 A1 WO 9313213A1
Authority
WO
WIPO (PCT)
Prior art keywords
gas
reaction vessel
microorganisms
microorganism
methane
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/JP1987/000156
Other languages
English (en)
Japanese (ja)
Inventor
Shinko Kitaura
Yoshimasa Takahara
Shiro Nagai
Naomichi Nishio
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Individual
Original Assignee
Individual
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from JP61054697A external-priority patent/JPS62215395A/ja
Priority claimed from JP61054696A external-priority patent/JPS62236489A/ja
Application filed by Individual filed Critical Individual
Priority to US07/093,497 priority Critical patent/US4921799A/en
Anticipated expiration legal-status Critical
Publication of WO1993013213A1 publication Critical patent/WO1993013213A1/fr
Ceased legal-status Critical Current

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Classifications

    • 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
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
    • 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
    • C12M21/00—Bioreactors or fermenters specially adapted for specific uses
    • C12M21/12—Bioreactors or fermenters specially adapted for specific uses for producing fuels or solvents
    • 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
    • C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/141—Feedstock

Definitions

  • the present invention relates to a method for producing a substance using a microorganism, and more particularly, to a novel method for biosynthesizing a target substance from a gaseous raw material using an immobilized microorganism.
  • the present invention plays an important role in the technical field of biotechnology such as fermentation industry, microbial industry, enzyme industry, and food industry.
  • the present invention can also be used extensively in the technical field of the chemical industry, because methane, hydrogen cyanide, acetylene and other various organic industrial chemicals can be produced using the methane and the like obtained by this method as a raw material. Is what is done.
  • a system for producing methane using an apparatus as shown in Fig. 2 has been developed. It consists of methanotrophs in a liquid medium containing supplementary nutrients such as nitrogen sources and inorganic salts.
  • carbon dioxide gas and hydrogen gas are forcibly supplied from the outside of the fermenter 13 into the liquid medium, and mechanically stirred by the stirring blades 14 (aeration-stirred fermenter 15) or draft.
  • the air bubble 19 from the vent hole 18 is atomized by the air tube 16 (bubble column type fermenter 17) to increase the gas-liquid interface, and at the same time, the air bubble is retained in the culture medium for a long time, thereby increasing the culture medium.
  • This system accelerates the rate of dissolution of carbon dioxide and hydrogen gas into the gas and causes a biochemical reaction by the methane-producing bacteria to obtain a product gas.
  • This system also converts methane from the raw material gas. However, it cannot be biosynthesized by a direct gas phase reaction, and as described later, it cannot be used industrially because of its drawbacks such as low methane production rate.
  • This method also does not biosynthesize the desired substance directly from the raw material gas by a gas phase reaction, and has the disadvantage that the production rate of the desired substance is low. I can not do such a thing.
  • FIG. 1 shows an example of an apparatus for carrying out the present invention.
  • Fig. 2 shows a conventional methane fermentation apparatus.
  • FIG. 3 illustrates an example of an apparatus for producing formic acid in the present invention.
  • the present invention has been made for the purpose of developing an industrial system for directly producing a target product from a gaseous substrate, and firstly uses the above-described aeration type or bubble column type fermenter. We focused on the system.
  • Aeration-type fermenters require large power for mechanical stirring.
  • the present invention has been made to solve the above-mentioned drawbacks and to develop an industrial production method for producing the target substance in a large amount and economically.
  • Reactor l contains the surface-treated microorganisms. Immobilization of microorganisms is carried out by a conventional method, and any of the carrier binding methods can be used.
  • microorganisms are immobilized in a spherical, cylindrical, granular or other suitable shape and then filled into the reactor 1, fixed directly to the reactor wall, or immobilized on the inner and / or outer surface.
  • a large number of hollow fibers with immobilized microorganisms are filled into the reactor, and one or more (porous) plates with immobilized microorganisms are vertically or horizontally filled into the reactor.
  • the above-mentioned molded and immobilized cells are filled into a small column and then filled into a large number of reactors to constitute a reactor.
  • any microorganism can be used as long as it produces the target product using a gaseous substrate.
  • a methane-producing bacterium of a type utilizing carbon dioxide gas, hydrogen gas, or the like as a substrate can be advantageously used, but the present invention is not limited to only these microorganisms. All microorganisms can be used as long as the target product can be biosynthesized using the substrate.
  • the methane-producing bacterium is a gram-negative methane-producing bacterium HU strain isolated from digested sludge at a sewage treatment plant in Hiroshima City (a strain preserved in the Nagai Laboratory, Faculty of Engineering, Hiroshima University; freely available for sale). ,
  • Methanosaliina used alone or in combination it can.
  • bacteria sources such as culture solutions, wet cakes, activated sludge, and digested sludge and use them in the present invention.
  • An aqueous solution 2 containing supplementary nutrients such as a nitrogen source and inorganic salts is sprayed, dropped or dropped from a squirt tube 4 onto a carrier on which organisms or microorganisms are fixed by a control valve 3. If necessary, these nutrient solutions may be stored in a carrier in advance.
  • aqueous solution When the nutrient solution used in Examples 1 to 3 is used as the aqueous solution to be supplied to the microorganisms in the present invention, methane gas is generated, and when the solution in Example 4 is used, formic acid is generated. It is. At the same time when the aqueous solution 2 is dropped, a substrate gas having an appropriate composition is supplied from the lower pipe 5 of the reactor via the regulating valve 6 and brought into contact with the microorganisms and nutrient solution fixed on the carrier to produce the desired product. Is generated. Since the type and composition of the substrate gas used as a raw material differ depending on the bacterium used, it is necessary to select an optimum one according to the bacterium used.
  • the H 2 / CO 2 ratio is preferably larger than 1.
  • a gas analyzer (not shown) is installed at the product gas outlet 7 to analyze the product gas, It is preferable to operate the control valve 6 provided at the inlet so as to adjust the mixing ratio of the substrate gas and / or the supply amount and the supply speed thereof to the optimum values for the production of methane.
  • the control valve 6 is controlled according to the data of the gas analyzer to supply the substrate in accordance with the substrate consumption rate of the microorganism.
  • Reactor 1 is surrounded by a jacket for heating or keeping the temperature warm, in which hot water or temperature-adjusted gas is flown, and a heating wire is provided to conduct biosynthesis reaction. May be promoted. Conversely, it is also possible to supply the substrate gas from above the reactor and take out the generated gas from below the reactor. Also, the aqueous solution that has fallen into the liquid collecting tank 8 is not discarded as it is but is passed from the liquid outlet 9 to the aqueous solution tank 2 via a pump and a pipe (not shown). Recycling further increases its economic efficiency.
  • the product is a water-soluble substance such as formic acid
  • it is dissolved in the aqueous solution and falls into the liquid collecting tank 8, so that the aqueous solution is withdrawn from the liquid outlet 9 to recover the water-soluble substance.
  • pressurizing the inside of the reactor can increase the gas solubility and increase the reaction rate. If the reactor is kept airtight and the base gas content in the reactor is adjusted to the optimal value for the immobilized microorganisms, the production rate of the target product can be maximized.
  • the target substance thus generated is a gas
  • the target substance is collected in the gas storage 10 through the generation gas outlet 7.
  • the target substance If is a water-soluble substance, it is recovered from the liquid collection tank 8.
  • Zeolite, foam brick, and inorganic foam (particle size: 7.1 to 12.6 mm) were used as carriers, and the HU strain, which had been isolated in the laboratory of Hiroshima University's Faculty of Engineering, was used as the carrier. It was fixed by the adsorption method.
  • the methane-producing bacteria immobilized on the carrier in this manner are filled in the reactor shown in Fig. 1 (reactor capacity: 75 mJi), and the gaseous substrate is obtained using the equipment shown in Fig. 1 under the following specifications. Methane fermentation was performed. That is, in the reactor 1 filled with the carrier on which the microorganisms are immobilized, the nutrient solution 2 shown in Table 1 is dropped from the upper part of the carrier from the ejection pipe 4 by the control valve 3 onto the surface of the carrier, and the lower part of the reactor is An appropriate flow of the substrate gas was supplied from the pipe 5 by the control valve 6, and the gas generated by the microorganisms on the carrier was obtained from the upper outlet 7. A jacket was provided around the reactor, and the temperature was adjusted to the optimal temperature (37 ° C) for microbial reaction by passing temperature-controlled water.
  • Example 3 Inorganic foam 0.604g-dry cell
  • Substrate gas supply rate 4760mfi / day
  • a suspension of HU strain dry cell concentration of 10.86 g / ⁇
  • the resulting solution containing formic acid and the unreacted gas were recovered.
  • 115 is a formic acid-containing solution outlet
  • 116 is a gas outlet
  • 117 is a temperature control water inlet at 32
  • 118 is a temperature control water outlet
  • 119 is a jacket.
  • Methyl viologen 7.5 mMol / fi Table 3 shows the fermentation conditions for formic acid. Under these conditions 32 to 2 weeks anti The results are shown in Table 3.
  • the formic acid was quantified by the method of Lang et al. (Lang E, Lang H., Z. Anal. Chem., 260, 8-10 (1972)), and the gas composition was gas chromatograph, gas chromatograph. The flow rate was determined by the stone membrane method.

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Sustainable Development (AREA)
  • General Chemical & Material Sciences (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Immunology (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

L'invention se rapporte à un procédé de fermentation pour produire un produit en fermentant un substrat gazeux au moyen de micro-organismes. Ce procédé consiste à retenir un micro-organisme immobilisé sur un support dans un récipient de réaction, à réintroduire une solution aqueuse pour qu'au moins une partie de la surface du micro-organisme s'humidifie, et à faire passer le substrat gazeux à travers les vides de l'aggrégat du micro-organisme, pour faire réagir directement le micro-organisme avec le substrat gazeux. Ainsi, du méthane, de l'acide formique ou similaires peuvent être efficacement biosynthétisés.
PCT/JP1987/000156 1986-03-14 1987-03-13 Procede de fermentation Ceased WO1993013213A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US07/093,497 US4921799A (en) 1986-03-14 1987-03-13 Fermentation method

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
JP61/54696 1986-03-14
JP61/54697 1986-03-14
JP61054697A JPS62215395A (ja) 1986-03-14 1986-03-14 発酵方法
JP61054696A JPS62236489A (ja) 1986-03-14 1986-03-14 メタンの製造方法

Publications (1)

Publication Number Publication Date
WO1993013213A1 true WO1993013213A1 (fr) 1993-07-08

Family

ID=26395495

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP1987/000156 Ceased WO1993013213A1 (fr) 1986-03-14 1987-03-13 Procede de fermentation

Country Status (1)

Country Link
WO (1) WO1993013213A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE4438023C1 (de) * 1994-10-25 1995-12-07 Buna Gmbh Verfahren zur Substratversorgung bei der Kultivierung von Mikroorganismen

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS58223496A (ja) * 1982-06-18 1983-12-26 Mitsubishi Heavy Ind Ltd メタン発酵装置

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS58223496A (ja) * 1982-06-18 1983-12-26 Mitsubishi Heavy Ind Ltd メタン発酵装置

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE4438023C1 (de) * 1994-10-25 1995-12-07 Buna Gmbh Verfahren zur Substratversorgung bei der Kultivierung von Mikroorganismen

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