Classifications

• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
  • C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
  • C12N11/04—Enzymes or microbial cells immobilised on or in an organic carrier entrapped within the carrier, e.g. gel or hollow fibres
• • 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
  • C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
• • 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
  • C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
  • C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
  • C12P7/06—Ethanol, i.e. non-beverage
• • 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
  • Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
  • Y02E50/00—Technologies for the production of fuel of non-fossil origin
  • Y02E50/10—Biofuels, e.g. bio-diesel

Definitions

• the present invention relates to a process in the microbial fermentation of an organic medium and to a microbial catalyst suitable for use in said process.
• Immobilized microorganisms have been used earlier for the preparation of i.a. ethanol.
• Cf. The immobilization of microbial cells, subcellular organelles and enzymes in calcium alginate gels" by M. Kierstan and C. Bucke, Bio- technol. Bioeng. 19, 387 (1977).
• Immobilization of a microorganism means that it is associated with a carrier material, e.g. polymeric materials, such as polyacrylamide or calcium alginate.
• the invention relates to a process in microbial fermentation of organic substrate, the fermentation being carried out with a microorganism that is immobilized in a carrier material together with a magnetically influenced substance so that the ratio between the weight of the micro organism and the weight of the magnetically influenced substance is within the range 5-500, preferably 10-200, an the microorganism and the fermented organic substrate being separated upon completion of the fermentation by means of a magnetic field.
• the separation can e.g. be effected by keeping the microbial catalyst (being understood to comprise the micro- organism immobilized in the carrier material together with a magnetically influenced substance) in the reactor by magnetic forces while the reactor is emptied.
• Another way of effecting separation is to catch the microbial catalyst outside the reactor and then return it to the reactor for use in the fermentation of the next substrate batch.
• Still another variant is to keep the microbial catalyst in the reactor by means of a magnetic field while the organic substrate continuously is allowed to flow through the reactor at an adjustable rate.
• One way is to extract continuously the reactor contents during the course of the fermentation with an organic solvent removing the fermentation product formed.
• the solvent must of course be adapted to the fermentation product.
• frequently occuring solvents are alcohols such as ethanol, octanol, decanol and dodecanol; esters such as dibutyl phthalate and tributyl phosphate; aliphatic and/or aromatic hydrocarbons; chlorinated hydrocarbons such as bromoethane, 2-bromopropane and dichloroethane.
• the very combination of immobilized microorganisms and solvent extraction is essential since immobilization protects the microorganisms from being harmfully influenced by the solvent used.
• the microbial catalyst is magnetically influenced the catalyst and the organic substrate can be separated magnetically.
• a magnetic field can keep the microbial catalyst in the reactor while the organic substrate containing the fermentation product is pumped to an extraction unit and thereupon is returned to the reactor.
• the principle of magnetic separation allows a rapid and safe circulation of the organic substrate for a long. time. If instead a filter disc is used as separator, the pores of the filter disc are easily clogged by the suspended particles present leading to operating interruptions
• Another way of keeping the concentration of the fermentation product low in the reactor is to boil off the same continuously from the organic medium in vacuum.
• the method of boiling off the fermentation product in vacuum from the organic substrate has been disclosed earlier for non-immobilized microorganisms (G.R. Cysewski and C.R. Wilke, Bio technol, Bioeng. 19 , 583 (1977)) but said technique has not been considered to be economically justifiable owing to the large amounts of energy required in order to pump away the carbon dioxide formed.
• the microbial catalyst according to the invention is charac- terized in that the microorganism is immobilized in a carrier material together with a magnetically influenced substance, the ratio between the weight of the microorganism and the weight of the magnetic substance being 5-500, pre- ferably 10-200.
• a magnetically influenced substance the ratio between the weight of the microorganism and the weight of the magnetic substance being 5-500, pre- ferably 10-200.
• the carrier material is suitably a polymer, such as calcium alginate, polyacrylamide, polymethacrylate and carragenate whereas the magnetically influenced substance (being understood to be a material attracted by a magnetic field) can be iron filings, magnetite, ferrite, etc., preferably ferrite powder.
• the microorganisms to be used can be of widely different types. Thus, it has been shown that fungi and bacteria well maintain their catalytic activity when applying the present invention but use can also be made of other kinds of microorganisms, such as virus, algae and protozoa.
• a microbial catalyst was prepared in accordance with Example 1, however with the exception that the ferrite powder used in Example 1 and coated with organic material was replaced by 2 grams of ferrite powder.
• a microbial catalyst prepared in accordance with Example 2 in an amount of 2 grams was mixed with 10 ml of 5% glucose solution containing 10 mM calcium chloride. The mixture was stirred in a vessel provided with a cover at 30o and the formation of ethanol was followed by means of gas chromatography.
• the corresponding non-immobilized microorganism as well as microorganism immobilized according to Example 2 but in the absence of ferrite were used.
• the magnetic catalyst is as effective as the non-magnetic one and almost as effective as free microorganism.
• the magnetic microbial catalyst was prepared by means of a strong permanent magnet, was washed with water while still attached to the magnet and then transferred to the next fermentation cycle. This separation could also be carried out in the presence of great amounts of solid particles. As solid particles use were made of sand, glas and wood chips, in the order mentioned, in amounts of 20% by volume.
• a 5% sugar solution in an amount of 10 ml was fermented with a microbial catalyst according to claim 1 (5 grams) in the presence of (test A) and in the absence of (test B) 50 ml of dodecanol.
• a microbial catalyst according to claim 1 5 grams
• the two fermen- tation processes proceeded equally fast but during the later stage of the fermentations the fermentation in test A proceeded about 20% fasterthan the fermentation in test B dependent on the beneficial effect of the removal of the ethanol from the aqueous phase.
• Example 5 A fermentation process similar to that of Example 5 was carried out but using decanol instead of dodecanol while tests A and B were carried out using a starting concentration of 7% by weight of ethanol. During the continued fer- mentation of glucose test A proceeded more than twice as fast as test B. This illustrates the importance of extracting ethanol at high ethanol concentrations.
• a microbial catalyst according to Example 2 was kept magnetically in a reactor of the volume 100 ml through which there was continuously pumped a 10% glucose solution or a 10% glucose solution containing nutrients (0.25% yeast extract, 0.025% NH 4 H 2 PO 4 , 0.0025% MgSO 4 ). The temperature was 30oC.
• a 10% glucose solution or a 10% glucose solution containing nutrients 0.25% yeast extract, 0.025% NH 4 H 2 PO 4 , 0.0025% MgSO 4 .
• the temperature was 30oC.
• the ethanol- yielding capacity as a function of time. In the presence of nutrients the catalyst according to the invention has a good ethanol-yielding capacity during a long time.
• a microbial catalyst according to Example 8 immobilized Zymomonas mobilis but without magnetic substance and free Zymomonas mobilis cells were compared with respect to the ethanol-yielding capacity according to Example 3.
• the capacity of the immobilized bacterium was not influenced nega tively by the incorporation of magnetic material in the carrier material and was about 75% of the capacity of the free bacterium.

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• Chemical & Material Sciences (AREA)
• Organic Chemistry (AREA)
• Life Sciences & Earth Sciences (AREA)
• Wood Science & Technology (AREA)
• Engineering & Computer Science (AREA)
• Zoology (AREA)
• Health & Medical Sciences (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Genetics & Genomics (AREA)
• Biochemistry (AREA)
• Microbiology (AREA)
• Biotechnology (AREA)
• General Engineering & Computer Science (AREA)
• General Health & Medical Sciences (AREA)
• Chemical Kinetics & Catalysis (AREA)
• General Chemical & Material Sciences (AREA)
• Biomedical Technology (AREA)
• Dispersion Chemistry (AREA)
• Mycology (AREA)
• Preparation Of Compounds By Using Micro-Organisms (AREA)
• Immobilizing And Processing Of Enzymes And Microorganisms (AREA)

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• 1980
  • 1980-08-22 WO PCT/SE1980/000215 patent/WO1981000575A1/en not_active Application Discontinuation
  • 1980-08-22 EP EP80901624A patent/EP0041065A1/en not_active Ceased
  • 1980-08-22 JP JP50193880A patent/JPS56501191A/ja active Pending
  • 1980-08-22 BR BR8009026A patent/BR8009026A/pt unknown
  • 1980-08-25 FI FI802671A patent/FI802671A/fi not_active Application Discontinuation
• 1981
  • 1981-04-23 DK DK182781A patent/DK182781A/da unknown

Non-Patent Citations (1)

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