KR20160124477A - Method for producing organic acid through organic acid fermentation by microorganism, and device for production of organic acid - Google Patents

Method for producing organic acid through organic acid fermentation by microorganism, and device for production of organic acid Download PDF

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KR20160124477A
KR20160124477A KR1020150055125A KR20150055125A KR20160124477A KR 20160124477 A KR20160124477 A KR 20160124477A KR 1020150055125 A KR1020150055125 A KR 1020150055125A KR 20150055125 A KR20150055125 A KR 20150055125A KR 20160124477 A KR20160124477 A KR 20160124477A
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organic acid
fermentation
microorganism
carbon dioxide
fermentation broth
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KR101812383B1 (en
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이태호
상병인
전재성
전병승
박효정
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한양대학교 산학협력단
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    • C12P7/00—Preparation of oxygen-containing organic compounds
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    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/04—Filters; Permeable or porous membranes or plates, e.g. dialysis
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/18—External loop; Means for reintroduction of fermented biomass or liquid percolate
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    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/20—Degassing; Venting; Bubble traps
    • 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
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/26—Means for regulation, monitoring, measurement or control, e.g. flow regulation of pH

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Abstract

The present method relates to a method for producing organic acid by microbial organic acid fermentation, and to an apparatus for performing the method. More specifically, the present invention relates to a method for producing organic acid by microbial organic acid fermentation comprising: a) a step of separating a fermentation reaction solution in which a fermentation reaction is carried out by microorganisms into the microorganisms and a fermented liquor that is a fermentation reaction output; b) a step of lowering the pH of the fermented liquor; c) a step of contacting the pH-lowered fermented liquor of the step b) with an organic solvent for organic acid extraction to extract organic acid from the pH-lowered fermented liquor; and d) a step of raising the pH of the fermented liquor from which the organic acid is extracted of the step c), and joining the pH-raised fermented liquor with the fermentation reaction solution in which the fermentation reaction is carried out by the microorganisms of the step a). The present invention also relates to an apparatus for performing the method. A method for producing organic acid according to the present invention is capable of producing the organic acid consistently and highly efficiently without lowering the production efficiency of the organic acid by organically controlling the pH at optimal conditions throughout the entire process. Furthermore, the method for producing organic acid according to the present invention is capable of offering expense saving, and is free from a salt accumulation problem in the reaction solution since a separate pH controlling agent is not additionally added.

Description

TECHNICAL FIELD [0001] The present invention relates to a method for producing an organic acid by microbial organic acid fermentation and an apparatus for producing the same,

The present invention relates to a method for producing an organic acid by microbial organic acid fermentation and an apparatus for carrying out the method.

Conventional microbial fermentation reactions are used to produce various organic acid or alcohol-based compounds. For example, various organic acids such as acetic acid, propionic acid, butyric acid, and succinic acid can be produced through microbial fermentation reaction, and various alcohol compounds such as ethanol and butanol can also be produced through a microbial fermentation reaction. The resulting organic acids or alcohol compounds can be used as platform chemicals in a variety of chemical processes and can be used to synthesize materials with higher value through subsequent catalytic processes.

On the other hand, the term "organic acid fermentation by microorganisms" refers to a process for producing various organic acids by incomplete oxidation of a carbon compound through a fermentation reaction with microorganisms. At this time, the concentration of the organic acid in the fermentation reaction solution Is increased. Thus, if the concentration of organic acid in the fermentation reaction solution increases, the pH of the fermentation reaction solution will be lowered, and the lowered pH will again be a factor for inhibiting the fermentation reaction of microorganisms.

Accordingly, a means for returning the lowered pH to a pH suitable for the fermentation reaction as the reaction progresses is required. To adjust the pH, a substance such as an alkali salt is added to artificially raise the pH of the lowered fermentation reaction solution , The added alkali salt reacts with the acid in the fermentation reaction liquid to form a salt, and the resulting salt is accumulated in the fermentation reaction liquid, and the volume of the fermentation reaction solution also increases.

Korean Patent Laid-Open Publication No. 10-2013-0020037 discloses an anaerobic digestion system capable of adjusting the pH. According to the present invention, an anaerobic digestion system capable of adjusting the pH of a digestive juice is provided without adding a buffer. Specifically, in an anaerobic digestion system including an acid fermentation tank, a digestive juice discharge line, a methane fermentation tank, a digestion gas discharge pipe, a CO 2 absorption tower, and a digestion gas supply pipe, digestion gas generated in an acid fermentation tank is introduced into the CO 2 absorption tower and it processed and reducing the CO 2 concentration is, by being fed to a methane fermentation tank for fire extinguishing gas CO 2 concentration decreases starting the anaerobic digestion system, capable of adjusting pH is the pH of the methane fermentation tank control.

On the other hand, as described above, in the fermentation reaction, when the pH is high, the activity of the microorganism is not inhibited. Therefore, when the organic acid produced by the fermentation reaction is extracted using an organic solvent or the like, Do.

Korean Patent Laid-Open Publication No. 10-2012-0025998 discloses a method for producing alkylbutyrate from a fermentation broth of a microorganism. Specifically, a fermentation broth is produced using a butyric acid producing strain, and then carbon dioxide Converting the butyrate salt to butyric acid, and extracting it with an extraction solvent, and the like.

However, the above-mentioned techniques require a device such as a separate CO 2 absorption tower for pH adjustment by CO 2 concentration control, and an artificial rise in pH or fermentation product extraction to prevent inhibition of microbial fermentation reaction But it does not disclose how to effectively control the pH in a continuous process leading to extraction of fermentation product from the fermentation reaction.

Korean Patent Publication No. 10-2013-0020037 Korean Patent Publication No. 10-2012-0025998

SUMMARY OF THE INVENTION Accordingly, the present invention has been made in view of the above problems, and it is an object of the present invention to provide a method for producing organic acids by microbial organic acid fermentation by adjusting the pH to an optimum condition organically throughout the process, A method of producing an organic acid by fermentation of an organic acid by a microorganism which can produce an organic acid with excellent efficiency and can further reduce the cost by eliminating the pH adjuster added separately and without the problem of salt accumulation in the reaction solution, And to provide a device for such use.

In order to solve the above problems,

A method for producing an organic acid by fermentation of an organic acid by a microorganism,

a) separating the fermentation reaction solution in which the fermentation reaction by the microorganism is performed into a fermentation solution which is the result of the fermentation reaction with the microorganism;

b) lowering the pH of the fermentation broth;

c) extracting the organic acid from the fermentation broth having the lowered pH by contacting the fermentation broth having the lowered pH in the step b) with an organic solvent for extracting organic acid; And

d) adding the fermentation broth having the increased pH to the fermentation broth in which the fermentation reaction with the microorganism in step a) is performed, after raising the pH of the fermentation broth from which the organic acid has been extracted in step c)

A method for producing an organic acid by microbial organic acid fermentation.

According to an embodiment of the present invention, the separation of the microorganism and the fermentation broth in step a) may be performed by passing the fermentation reaction solution through a membrane separation device.

According to another embodiment of the present invention, the membrane separation device may be a ceramic material film or a polymer material film having pores having a diameter of 0.2 to 0.5 μm.

According to another embodiment of the present invention, the pH drop in step b) may be performed by gas-liquid contacting the carbon dioxide gas with respect to the fermentation broth.

According to another embodiment of the present invention, the gas-liquid contacting is performed by stirring the carbon dioxide gas at a pressure of 5 to 50 bar and a temperature of 4 to 40 DEG C for 10 to 20 minutes with the fermentation broth at 500 rpm or less Lt; / RTI > conditions.

According to another embodiment of the present invention, the carbon dioxide gas may be a carbon dioxide gas generated from the fermentation reaction in the step a).

According to another embodiment of the present invention, the pH of the fermentation broth after the pH drop of step b) may be 4 to 5.

According to another embodiment of the present invention, the organic solvent in step c) may be selected from the group consisting of butyl butyrate, dodecanol, oleyl alcohol, and mixtures thereof.

According to another embodiment of the present invention, the pH increase in step d) may be performed by degassing the carbon dioxide from the fermentation liquid whose pH has been lowered by gas-liquid contact with carbon dioxide in step b).

According to another embodiment of the present invention, the degassing of the carbon dioxide can be performed by applying a pressure of 100 mmHg to 760 mmHg and a temperature condition of 20 ° C to 30 ° C to the fermentation broth.

According to another embodiment of the present invention, the pH of the fermentation broth after the increase of the pH of step d) may be 5.5 to 6.5.

According to another embodiment of the present invention, the organic acid may be selected from the group consisting of butyric acid, hexanoic acid, and mixtures thereof.

According to another embodiment of the present invention, the microorganism may be a single microorganism or a mixed microorganism producing an organic acid.

According to another embodiment of the present invention, the fermentation broth in contact with the organic solvent for extracting organic acid in step c) may be separated into an organic solvent containing the organic acid and a fermentation broth from which the organic acid is extracted through an oil-water separator.

Meanwhile, the present invention also relates to an apparatus for carrying out the above method,

An organic acid production fermenter for containing a microorganism culture liquid for performing organic acid fermentation;

A membrane separation device for separating the culture medium transferred from the organic acid production fermenter into a fermentation broth and a microorganism;

A microorganism return line for carrying the microorganism from the membrane separation apparatus to the organic acid production fermenter;

An extractor for extracting the organic acid from the fermentation broth transferred from the membrane separation apparatus using an organic solvent;

An oil-water separator for separating a mixture of the fermentation broth and the organic solvent transferred from the extractor;

A carbon dioxide inflow line for transferring the carbon dioxide gas produced by the organic acid production fermenter to the extractor;

A fermentation liquid transfer line for transferring the fermentation liquid from which the organic acid has been extracted from the oil water separator to the organic acid production fermenter; And

The present invention also provides an apparatus for producing organic acid by microbial organic acid fermentation, which comprises a carbon dioxide degassing tank installed in the middle of the fermentation liquid return line for performing degassing reaction of carbon dioxide.

According to an embodiment of the present invention, a surge tank may be further provided between the extractor and the oil-water separator to supply a mixed solution of the fermentation broth and the organic solvent.

According to another embodiment of the present invention, the carbon dioxide degassing vessel may further include a stirrer or a vacuum pump for facilitating the degassing of the carbon dioxide.

According to the present invention, it is possible to produce organic acids continuously and with high efficiency without deteriorating organic acid production efficiency by adjusting the pH to an optimum condition organically throughout the entire process, and further, there is no pH adjusting agent added separately, And it is possible to provide an organic acid production method free from the problem of salt accumulation in the reaction solution.

1 is a schematic process diagram of a method for producing organic acids by microbial organic acid fermentation according to the present invention.
2 is a diagram showing an example of a device design diagram for manufacturing the device according to the present invention.
Fig. 3 shows a photograph of a device actually manufactured in accordance with the design of Fig.
FIG. 4 is a graph showing the extraction efficiency according to pH when organic acids are extracted from a non-fermented culture solution using pH control according to a conventional method.
FIG. 5 is a graph showing the difference in extraction efficiency depending on the extraction time, while varying the extraction time for the culture medium having the pH adjusted to 4.5, by extracting organic acids from the non-fermented culture using the pH control according to the conventional method Fig.
6 is a graph showing extraction efficiency according to pressure conditions applied to a high-pressure extractor in extracting organic acids from a non-fermented culture liquid through pH control using carbon dioxide in the extraction step according to the present invention.
FIG. 7 is a graph showing extraction efficiency according to pressure conditions applied to a high-pressure extractor in extracting organic acids from a fermented culture liquid through pH control using carbon dioxide in the extraction step according to the present invention. FIG.
8 is a schematic view showing a process flow chart for the microbial membrane separation process according to the third embodiment.
9 is a graph showing the concentration of butyric acid and the concentration of butyric acid in an extraction solvent over time according to Example 3 when fermentation of microbial organic acids and extraction of the produced organic acid are performed.

Hereinafter, the present invention will be described in more detail with reference to the drawings and examples.

FIG. 1 shows a schematic process diagram of a method for producing organic acids by microbial organic acid fermentation according to the present invention.

The method according to the invention,

a) separating the fermentation reaction solution in which the fermentation reaction by the microorganism is performed into a fermentation solution which is the result of the fermentation reaction with the microorganism;

b) lowering the pH of the fermentation broth;

c) extracting the organic acid from the fermentation broth having the lowered pH by contacting the fermentation broth having the lowered pH in the step b) with an organic solvent for extracting organic acid; And

d) bringing the pH of the fermentation broth in which the organic acid extracted in the step c) is increased to the pH of the fermentation broth and joining the fermentation broth with the fermentation reaction solution in which the fermentation reaction with the microorganism in the step a) is performed.

Referring to FIG. 1, a fermentation reaction by a microorganism is performed in an organic acid production fermenter 110, and a microorganism and a microorganism culture solution for performing organic acid fermentation are accommodated in the fermenter 110. The process according to the present invention is suitable for producing various organic acids, but organic acids selected from the group consisting of butyric acid, hexanoic acid and mixtures thereof can be produced according to the present invention. In order to produce the organic acid, it is possible to use the organic acid producing microorganism known in the art alone, but it is also possible to use a mixed microorganism of such organic acids.

In the organic acid producing fermenter 110, a microorganism that performs an organic acid fermentation and a fermentation liquid, which is a result of fermentation by the microorganism, are mixed. In the present invention, a process of separating the mixed liquid into a microorganism and a fermentation liquid is performed , Which is performed in the microbial / fermentor separator 120 shown in FIG.

The reason why the microorganism and the fermentation broth are separated from each other in the present invention is that the organic acid produced is accumulated in the organic acid production fermenter 110 as the microbial fermentation process proceeds, The pH of the fermentation reaction liquid in the organic acid production fermenter 110 is lowered, and the lowered pH is again a factor for inhibiting the fermentation reaction of the microorganism. On the other hand, low pH may be a factor for inhibiting the fermentation reaction of microorganisms. However, in the process of extracting the organic acid produced later, the extraction reaction may facilitate the extraction reaction. Therefore, in the present invention, the microorganism and the fermentation broth are separated from each other, and the microorganism is returned to the organic acid production fermenter 110. The fermentation broth is further reduced in pH for extracting organic acid, and then the extraction process is performed. And returned to the fermenter (110). As a result, according to the method of the present invention, the pH of the fermentation reaction liquid in the organic acid production fermenter 110 can be prevented from being lowered, so that the fermentation reaction of the microorganism can be prevented from being inhibited, and at the same time, The organic acid can be extracted more easily in the organic acid extraction process by the organic solvent.

Meanwhile, in the present invention, the membrane separation apparatus 120 may be used for separating the microorganism from the fermentation broth in step a). The membrane separator 120 separates microorganisms from the fermentation reaction liquid to return only the microorganisms to the organic acid production fermenter 110. For example, the membrane separator 120 may be a membrane made of a ceramic material having pores having diameters of 0.2 μm to 0.5 μm Or a film made of a polymer material can be used.

Subsequently, in the step a), the fermentation liquid separated from the microorganism is subjected to an additional pH-lowering step. This is a process for facilitating organic acid extraction by an organic solvent in the future, as described above, and may be performed, for example, by gas-liquid contacting a carbon dioxide gas with respect to the fermentation broth. In the present invention, the pH lowering step in step b) can be performed by gas-liquid contacting the carbon dioxide gas with the fermentation liquid rather than adding a separate pH lowering agent. As a result, the carbon dioxide gas The pH of the fermentation broth is lowered. Specifically, the gas-liquid contact can be performed by contacting the carbon dioxide gas at a pressure of 5 bar to 50 bar and a temperature of 4 ° C to 40 ° C for 10 minutes to 20 minutes under stirring conditions of 500 rpm or less with the fermentation broth Liquid contact reactor and may be temporarily stored in the reservoir 130 after passing through the membrane separator 120 from the organic acid production fermenter 110 and then into a high pressure extractor 140 for extracting organic acid Or may be carried out and carried out. The reason why the high-pressure extractor 140 is used in the present invention is to facilitate dissolution of the carbon dioxide in the fermentation broth.

On the other hand, it is also possible to supply external carbon dioxide to lower the pH of the fermentation broth. Preferably, the carbon dioxide gas may be carbon dioxide gas generated from the fermentation reaction in the step a). In this case, it is possible to effectively recycle the carbon dioxide produced in the fermentation reaction for the purpose of lowering the pH of the fermentation broth. By this process, the pH of the fermentation broth into which carbon dioxide is injected can be lowered to 4 to 5 levels. The pH level is an extremely advantageous range for extracting the target organic acid using an organic solvent. Specifically, the solvent that can be used for the extraction of the organic acid includes, but is not limited to, butylbutyrate, dodecanol, oleyl alcohol, And mixtures thereof can be used.

1, the extraction organic solvent flowing into the high pressure extractor 140 flows from the first extraction solvent reservoir 160. The extracted extraction solvent is subjected to an organic acid extraction process in the high pressure extractor 140, The extraction solvent is separated into the fermentation liquid and the extraction solvent from which the organic acid is extracted through the oil-water separator 150, and the separated extraction solvent is transferred to the second extraction solvent reservoir 170. The extraction solvent stored in the second extraction solvent reservoir, Is an extraction solvent containing organic acids. Therefore, the organic acid is separated from the extraction solvent through a separate catalytic process, and then the extraction solvent in which the organic acid is separated is transferred to the first extraction solvent reservoir 160 again. Those skilled in the art will readily understand how to separate organic acids using an extraction solvent and separate the fermentation broth and extraction solvent using an oil-water separator.

In step d), in the present invention, the fermentation liquid extracted from the organic acid separated from the oil water separator 150 is merged with the fermentation reaction solution in step a), that is, As described above, since the pH of the fermentation broth is lowered by the microorganisms, the pH of the fermentation broth must be increased before the fermentation broth is returned to the fermentation broth.

Preferably, the pH increase in step d) can be performed by degassing the carbon dioxide from the fermentation broth whose pH has been lowered by gas-liquid contact with carbon dioxide in step b), because the fermentation broth can be removed from the carbon dioxide degassing bath (180), and applying a pressure of 100 mmHg to 760 mmHg to the fermentation broth and a temperature condition of 20 ° C to 30 ° C. Thus, the pH of the fermentation broth in which carbon dioxide has been degassed may be 5.5 to 6.5.

As a result, the method of producing organic acid by microbial organic acid fermentation according to the present invention can produce the target organic acid with excellent efficiency by repeating the above-described steps a) to d).

Meanwhile, the present invention also relates to an apparatus for carrying out the above method,

An organic acid production fermenter for containing a microorganism culture liquid for performing organic acid fermentation;

A membrane separation device for separating the culture medium transferred from the organic acid production fermenter into a fermentation broth and a microorganism;

A microorganism return line for carrying the microorganism from the membrane separation apparatus to the organic acid production fermenter;

An extractor for extracting the organic acid from the fermentation broth transferred from the membrane separation apparatus using an organic solvent;

An oil-water separator for separating a mixture of the fermentation broth and the organic solvent transferred from the extractor;

A carbon dioxide inflow line for transferring the carbon dioxide gas produced by the organic acid production fermenter to the extractor;

A fermentation liquid transfer line for transferring the fermentation liquid from the oil water separator to the organic acid production fermenter; And

The present invention also provides an apparatus for producing organic acid by microbial organic acid fermentation, which comprises a carbon dioxide degassing tank installed in the middle of the fermentation liquid return line for performing degassing reaction of carbon dioxide.

FIG. 2 shows an example of a device design diagram for manufacturing the device according to the present invention, and FIG. 3 shows a photograph of a device actually manufactured according to this design scheme. In the apparatus according to the present invention, the organic acid production fermenter, the membrane separation device (the organic acid production fermenter and the membrane separation device in the device design diagram of FIG. 2 are not shown), the organic acid production fermenter, A microbial return line, an extractor 240, an oil water separator 250, a carbon dioxide inflow line, a fermentation liquid return line, a first extraction solvent reservoir 260, a second extraction The description of the solvent reservoir 270 and the carbon dioxide degassing vessel 280 is the same as in the method according to the present invention described above.

2, a surge tank 290 may be further provided between the extractor 240 and the oil-water separator 250 to supply a mixed solution of the fermentation broth and the organic solvent. have. Particularly, when a high pressure is applied to the extractor 240 in order to smoothly dissolve carbon dioxide, the surge tank needs to use a pump for transferring the fluid from the extractor 240 to the oil water separator 250 In this case, a separate surge tank 290 may be required for smooth fluid transfer.

The carbon dioxide degassing vessel 280 may further include a stirrer and / or a vacuum pump to facilitate the degassing of the carbon dioxide.

EXAMPLES Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are intended to assist the understanding of the present invention and should not be construed as limiting the scope of the present invention.

Example

Comparative Example  1. According to the conventional method pH  Fermented through control Culture medium From organic acid extraction

Butyric acid was added at a concentration of 20 g / L into the non-fermentation broth at a concentration of 20 g / L, and the pH of the broth was adjusted to 4.0 and 4.5, respectively, using a sodium hydroxide solution. 5.0 and 5.5, respectively. 10 ml of each culture solution and 10 ml of butylbutyrate as an extraction solvent were added at room temperature and normal pressure, and the mixture was stirred at a speed of 700 rpm using a magnetic stirrer to completely mix the culture medium and the extraction solvent. After the extraction reaction, the mixture solution was allowed to stand for 5 minutes or more to separate the extraction solvent and the culture solution. The concentration of butyric acid present in the culture broth before and after the extraction was analyzed and the extraction efficiency was calculated by the following equation: (concentration of butyric acid before extraction - concentration of butyric acid after extraction) / (concentration of butyric acid before extraction) × 100. Respectively.

On the other hand, the extraction experiment was carried out according to the same method as above, except that the extraction time was changed to 10 minutes, 20 minutes, 30 minutes and 40 minutes for the culture medium having pH adjusted to 4.5, The results are shown in Fig. Referring to FIG. 5, it can be seen that, at an extraction time of 10 minutes or more, no improvement in extraction efficiency is observed with an increase in extraction time.

Example  1. Using carbon dioxide in the extraction step pH  Organic acid extraction from non-fermented culture medium

Butyric acid was added at a concentration of 20 g / L to the non-fermentation broth having a sugar concentration of 20 g / L, and the pH of the culture broth was adjusted to 6.0. 150 ml of the culture solution and 150 ml of butylbutyrate as an extraction solvent were put into a high-pressure extractor according to the present invention, and carbon dioxide was supplied to apply pressure of 10 bar, 20 bar, 30 bar, 40 bar and 50 bar, respectively. The temperature control was not performed separately, and the room temperature condition was maintained. Thereafter, the mixture was stirred for 10 minutes at a speed of 700 rpm using an impeller mounted on a high-pressure extractor so that the culture solution and the extraction solvent were completely mixed. After the extraction reaction, the mixture solution was allowed to stand for 5 minutes or more to separate the extraction solvent and the culture solution. The concentration of butyric acid present in the culture broth before and after the extraction was analyzed and the extraction efficiency was calculated by the following formula: (concentration of butyric acid before extraction - concentration of butyric acid after extraction) / (concentration of butyric acid before extraction) × 100. In FIG. 6, the extraction efficiency according to the pressure condition applied to the high-pressure extractor is shown in a graph. Referring to FIG. 6, it can be seen that the extraction efficiency increases as the pressure increases.

Example  2. In the extraction step, pH  From the fermented broth through control, organic acid extraction

A 2 L anaerobic fermenter was charged with 1.5 L of culture medium (glucose concentration 60 g / L) and clostridium tiobutiricum Tyrobutyricum ) were inoculated. Culture conditions were maintained at 37 ° C, pH 6.0, and cultured for 24 hours. After the fermentation, the microorganisms were separated from the fermentation broth and the concentration of butyric acid was measured to be 23 g / L in the fermentation broth filtered with microorganisms, and the pH was 6.0. 150 ml of the filtered fermentation broth and 300 ml of butylbutyrate as an extraction solvent were put into a high-pressure extractor according to the present invention, and carbon dioxide was supplied to apply pressure of 10 bar, 20 bar, 30 bar, 40 bar and 50 bar, respectively. The temperature control was not performed separately, and the room temperature condition was maintained. Thereafter, the mixture was stirred for 10 minutes at a speed of 700 rpm using an impeller mounted on a high-pressure extractor so that the culture solution and the extraction solvent were completely mixed. After the extraction reaction, the mixture solution was allowed to stand for 5 minutes or more to separate the extraction solvent and the culture solution. The concentration of butyric acid present in the culture broth before and after the extraction was analyzed and the extraction efficiency was calculated by the following formula: (concentration of butyric acid before extraction - concentration of butyric acid after extraction) / (concentration of butyric acid before extraction) × 100. In FIG. 7, the extraction efficiency according to the pressure condition applied to the high-pressure extractor is shown in a graph. Referring to FIG. 7, it can be seen that the extraction efficiency increases as the pressure increases.

Example  3. Microbial fermentation process through membrane separation process and carbon dioxide high pressure extraction process according to the present invention

A 2 L anaerobic fermenter was charged with 1.5 L of culture medium (sugar concentration 80 g / L) and clostridium tirobutiricum Tyrobutyricum ) were inoculated. The fermentation process was carried out at 37 ° C and pH 6.0. When the fermentation proceeded for 11 hours, the microorganisms in the 300 ml volume culture were filtered using a pore having an average diameter of 0.45 μm and a plate membrane having a surface area of 0.1 m 2 . At this time, the operating conditions of the flat plate membrane were as follows.

The fermented culture was fed to the plate membrane at a rate of 500 ml / min (flow rate of 300 L / m 2 / hr), and the microorganisms were recovered at a rate of 480 ml / min (flow rate of 288 L / m 2 / hr) , And the filtered culture was obtained at a rate of 20 ml per minute (flow rate of 12 L / m 2 / hr). For reference, FIG. 8 schematically shows a process flow chart for the microbial membrane separation process according to the third embodiment. In this embodiment, a flat membrane having a surface area of 0.1 m 2 is used, but the use of a flat membrane having a larger surface area can increase the volume of the culture liquid that can be treated. The recovered microorganisms were transported to a fermenter and used for culturing, and the subsequent organic acid extraction process was performed only on the filtered culture liquid as described below.

A 600 ml volume of extraction solvent (butyl butyrate) was preliminarily fed into the high pressure extractor and the pressure was increased to 50 bar by feeding the fermentation gas containing carbon dioxide. The temperature control was not performed separately, and the room temperature condition was maintained. Thereafter, 300 ml of the culture liquid filtered by the above procedure was supplied to the high-pressure extractor using a high-pressure pump.

The membrane process and the extraction process described above were maintained for 26 hours. As a result, no fouling phenomenon occurred inside the membrane. In the high-pressure extractor, the culture fluid circulation was performed at a rate of 1200 ml per hour (20 ml per minute) It went on. At this time, the extraction solvent was not circulated separately. After the extraction, the culture solution was re-fed to the fermenter through the degassing of carbon dioxide.

FIG. 9 shows the concentration of butyric acid and the concentration of butyric acid in the extraction solvent over time. From FIG. 9, it can be seen that the continuous production of organic acid and the continuous extraction of the produced organic acid by extraction solvent are performed. In this example, the volume of the culture medium and the extraction solvent were set to 1.5 L and 600 ml, respectively, but higher extraction efficiency could be achieved when the relative amount of the extraction solvent was increased or circulated.

Claims (17)

A method for producing an organic acid by fermentation of an organic acid by a microorganism,
a) separating the fermentation reaction solution in which the fermentation reaction by the microorganism is performed into a fermentation solution which is the result of the fermentation reaction with the microorganism;
b) lowering the pH of the fermentation broth;
c) extracting the organic acid from the fermentation broth having the lowered pH by contacting the fermentation broth having the lowered pH in the step b) with an organic solvent for extracting organic acid; And
d) adding the fermentation broth having the increased pH to the fermentation broth in which the fermentation reaction with the microorganism in step a) is performed, after raising the pH of the fermentation broth from which the organic acid has been extracted in step c)
Wherein the organic acid is fermented by a microorganism. The method according to claim 1, wherein the separation of the microorganism from the fermentation broth in step a) is performed by passing the fermentation reaction solution through a membrane separation device and filtering the microorganism. The method for producing organic acid by fermentation of microbial organic acids according to claim 2, wherein the membrane separation device is a film of a ceramic material or a polymer material having pores having a diameter of 0.2 to 0.5 m. The method according to claim 1, wherein the pH drop in step b) is carried out by bringing carbon dioxide gas into gas-liquid contact with the fermentation broth. 5. The method according to claim 4, wherein the gas-liquid contact is carried out by contacting the carbon dioxide gas with the fermentation broth at a pressure of 5 bar to 50 bar and at a temperature of 4 DEG C to 40 DEG C for 10 minutes to 20 minutes Wherein the microbial organic acid is fermented by a microorganism. The method of claim 4, wherein the carbon dioxide gas is carbon dioxide gas generated from the fermentation reaction in the step a). The method according to claim 1, wherein the pH of the fermentation broth after the pH reduction in step b) is 4 to 5. The method according to claim 1, wherein the organic solvent in step c) is selected from the group consisting of butyl butyrate, dodecanol, oleyl alcohol, and mixtures thereof. [7] The method according to claim 4, wherein the pH increase in step d) is carried out by degassing the carbon dioxide from the fermentation broth whose pH has been lowered by gas-liquid contact with carbon dioxide in step b) Production method. [Claim 11] The method according to claim 9, wherein the degassing of the carbon dioxide is performed by applying a pressure of 100 mmHg to 760 mmHg and a temperature condition of 20 [deg.] C to 30 [deg.] C to the fermentation broth. The method according to claim 1, wherein the pH of the fermentation broth after raising the pH of step (d) is 5.5 to 6.5. The method according to claim 1, wherein the organic acid is selected from the group consisting of butyric acid, hexanoic acid, and mixtures thereof. [Claim 3] The method according to claim 1, wherein the microorganism is a sole microorganism or a mixed microorganism that produces an organic acid. The method according to claim 1, wherein the fermentation liquid contacted with the organic solvent for extracting organic acid in step c) is separated into an organic solvent containing the organic acid and a fermentation liquid from which the organic acid is extracted through an oil water separator. Method of Producing Organic Acid by. An organic acid production fermenter for containing a microorganism culture liquid for performing organic acid fermentation;
A membrane separation device for separating the culture medium transferred from the organic acid production fermenter into a fermentation broth and a microorganism;
A microorganism return line for carrying the microorganism from the membrane separation apparatus to the organic acid production fermenter;
An extractor for extracting the organic acid from the fermentation broth transferred from the membrane separation apparatus using an organic solvent;
An oil-water separator for separating a mixture of the fermentation broth and the organic solvent transferred from the extractor;
A carbon dioxide inflow line for transferring the carbon dioxide gas produced by the organic acid production fermenter to the extractor;
A fermentation liquid transfer line for transferring the fermentation liquid from the oil water separator to the organic acid production fermenter; And
And a carbon dioxide degassing tank installed in the middle of the fermentation liquid return line for performing degassing reaction of carbon dioxide. The organic acid production apparatus according to claim 15, further comprising a surge tank between the extractor and the water-oil separator for constantly supplying a mixture of the fermentation broth and the organic solvent. . 16. The apparatus for producing organic acid by fermenting microbial organic acids according to claim 15, wherein the carbon dioxide degassing vessel is further provided with a stirrer or a vacuum pump for facilitating the degassing of the carbon dioxide.
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Citations (2)

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KR20120025998A (en) 2010-09-08 2012-03-16 에스케이이노베이션 주식회사 Method for preparing alkylbutyrate from microbial broth
KR20130020037A (en) 2011-08-18 2013-02-27 장희현 Anaerobic digestion system

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KR100329019B1 (en) 1999-04-13 2002-03-18 윤덕용 Method for Manufacturing Organic Acid by High-Efficiency Fermentation

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20120025998A (en) 2010-09-08 2012-03-16 에스케이이노베이션 주식회사 Method for preparing alkylbutyrate from microbial broth
KR20130020037A (en) 2011-08-18 2013-02-27 장희현 Anaerobic digestion system

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