WO2021015587A1 - Appareil de production de biotensioactif et procédé de production de biotensioactif l'utilisant - Google Patents

Appareil de production de biotensioactif et procédé de production de biotensioactif l'utilisant Download PDF

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Publication number
WO2021015587A1
WO2021015587A1 PCT/KR2020/009785 KR2020009785W WO2021015587A1 WO 2021015587 A1 WO2021015587 A1 WO 2021015587A1 KR 2020009785 W KR2020009785 W KR 2020009785W WO 2021015587 A1 WO2021015587 A1 WO 2021015587A1
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Prior art keywords
foam
air
bio
culture
bubbles
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PCT/KR2020/009785
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English (en)
Korean (ko)
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조양래
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주식회사 프록스엔렘
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Priority claimed from KR1020190090419A external-priority patent/KR102240151B1/ko
Priority claimed from KR1020200083635A external-priority patent/KR102432456B1/ko
Application filed by 주식회사 프록스엔렘 filed Critical 주식회사 프록스엔렘
Publication of WO2021015587A1 publication Critical patent/WO2021015587A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • 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
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/04—Apparatus for enzymology or microbiology with gas introduction means
    • C12M1/06—Apparatus for enzymology or microbiology with gas introduction means with agitator, e.g. impeller
    • 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
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/21—Froth suppressors
    • 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
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/34—Measuring or testing with condition measuring or sensing means, e.g. colony counters

Definitions

  • the present invention relates to an apparatus for producing a bio-surfactant and a method for producing a bio-surfactant using the same, and more specifically, to a culture unit for culturing a microorganism; A foam sensing unit positioned at the top of the culture unit and configured to sense a bubble generated from the culture unit; A first air injection tube for injecting air into the culture solution in the culture unit; A foam discharge unit through which the bubbles generated from the culture unit are discharged; A second air inlet tube for injecting air from the upper part of the culture unit toward the culture solution to dispel bubbles generated in the culture process to increase density; And a foam collection unit connected to the foam discharge unit and configured to collect the generated foam.
  • a bio-surfactant production device comprising: a bio-surfactant production device and a bio-surfactant production device comprising controlling the operation of the first and second air injection pipes according to bubble sensing of the bubble sensing unit. It relates to a method for producing a surfactant.
  • Bio-surfactants are natural surfactants present in animals and plants, such as lecithin, saponin, and bile acids, and surfactants produced by metabolism of microorganisms.
  • Surfactin a bio-surfactant, is known to be the most powerful material among bio-surfactants.
  • Bacillus subtilis include Bacillus Belle Zen sheath (B.velezensis), Bacillus amyl kwipe sieon Lowry's (B. amyloliquefaciens) to produce the seopaek tin (surfactin) bio-surfactant (biosurfactants) consisting of a lipid and an amino acid, Bacillus piece nipo miss (B licheniformis ), B. mojavensis , B. pumilus , and B. subtilis are known (Chen et al., 2015).
  • Surfactin is known to exhibit strong antiviral, antifungal, and antimicroplasma effects, and among them, antibiotic efficacy is the most known. Surfactin is in high demand in the pharmaceutical industry, environmental purification industry, cosmetics industry, etc. because it exhibits various effects as described above, but it has not been largely successful in commercialization because it is difficult to mass-produce due to low production efficiency.
  • surfactin has a strong emulsifying function, but it is used only as an expensive cosmetic and drug additive because its production efficiency is low and its production cost is high when it is used as a kitchen detergent, laundry detergent, and bath detergent, which are mainly used in everyday life.
  • synthetic detergents which are the cause of environmental pollution, into eco-friendly detergents
  • various Bacillus bacteria culture devices have been devised to increase the production efficiency of surfactin and lower the production cost. Became.
  • Korean Registered Patent No. 1139702 relates to a microbial complex culture device.
  • Korean Patent Registration No.0737709 uses a silicone foam control agent to prevent bubbles from occurring by causing a malfunction of the incubator or preventing the overflow of the incubator by attaching bubbles generated during cultivation of microorganisms to various sensors.
  • Korean Patent Registration No. 0737709 Korean Patent Registration No. 1120093 is a culture system that passively collects bubbles generated during microbial cultivation, and uses a passive method of connecting the bubble storage tank to the culture tank so that the bubbles generated in the culture tank pass to the bubble storage tank through a connection pipe. Use.
  • the culture system has a problem that the culture solution is moved to the bubble storage tank together with bubbles due to the increased pressure by the narrow connection pipe, and the loss of the culture solution is severe.Even if air is continuously injected, the growth efficiency of microorganisms is increased or the surfactant It is difficult to improve your productivity.
  • a bio-surfactant production device equipped with a foam sensing unit, a first air injection pipe, a second air injection pipe, and a foam discharge unit
  • foam By providing a sensing unit, a first air injection pipe and a second air injection pipe, mass production of surfactants is possible and bubbles can be discharged because only high-density bubbles can be moved to the foam collection unit by controlling the amount of bubbles and concentrating the bubbles.
  • the main object of the present invention is to control the amount of foam and concentrate the foam to move only high-density foam to the foam collection unit, so that mass production of surfactant is possible, and the foam overflows or the culture solution is discharged with the foam.
  • the main object of the present invention is to control the amount of foam and concentrate the foam to move only high-density foam to the foam collection unit, so that mass production of surfactant is possible, and the foam overflows or the culture solution is discharged with the foam.
  • Another object of the present invention is to provide a method for producing a bio-surfactant using the bio-surfactant production device.
  • the present invention provides a culture unit for culturing microorganisms, a foam sensing unit positioned at the top of the culture unit and for sensing bubbles generated from the culture unit, and injecting air into the culture solution in the culture unit.
  • a foam collection unit connected to the foam discharge unit and configured to collect generated bubbles, wherein the first and second air injection pipes are controlled according to the foam sensing unit. It provides a bio-surfactant production device characterized by.
  • the present inventors reduce the loss of the culture solution by reducing the pressure due to the movement of the bubbles through the bubble discharge unit, and control the operation of the first and second air injection pipes through the bubble sensing unit to control the amount of bubbles and By preventing collection in this storage tank, a bio-surfactant production apparatus was invented that could efficiently produce a bio-surfactant.
  • the first air injection pipe continuously injects air into the foam sensing unit until the foam is sensed, and after the foam is sensed, air is supplied at intervals of 1 to 10 minutes. It is characterized in that air is periodically injected by inflow and blocking.
  • the amount of air introduced into the first air injection pipe is 2.5 to 10 VVM until the foam is sensed, and 0.5 to 2.0 VVM after the foam is sensed.
  • the second air injection pipe when the foam is sensed in the foam sensing unit, the second air injection pipe is operated or blocked.
  • the second air injection pipe when the foam sensing unit senses the foam once, the second air injection pipe is operated (on), so that air is introduced in an amount of 0.5 to 1.5 VVM, and the foam is sensed twice.
  • the amount of air introduced into the second air injection pipe is increased to 2 to 10 VVM, and when the bubbles are sensed three times, the second air injection pipe is blocked (off).
  • the second air injection pipe is in the form of a spray.
  • the bubble discharge part is configured in a bottleneck shape, and the diameter of the upper end of the bubble discharge part is wider than the diameter of the lower end of the bubble discharge part through which bubbles are introduced.
  • the diameter of the upper end of the foam discharge part is 2 to 10 times wider than the diameter of the lower end of the foam discharge part into which the foam is introduced.
  • the culture unit is provided in a culture tank for culturing microorganisms, a propeller for agitation of a culture solution for agitating the culture solution, and a low density produced by being provided on the inside of the culture tank It characterized in that it comprises a propeller for removing bubbles for removing bubbles, and a control sensor provided inside the culture tank and for controlling the state of the culture solution.
  • the air injection unit is connected to an air pump equipped with a pressure gauge for injecting air, and is connected to the air pump and is connected to the inside of the culture tank and the bubble discharge unit on the other side connected to the air pump.
  • An air movement pipe for moving the introduced air, a first valve for controlling air inflow to the first air injection pipe, and a second valve for controlling air inflow to the second air injection pipe.
  • the foam collection unit is connected to the upper portion of the foam discharge unit, and is connected to a foam transfer pipe for moving the concentrated foam through the foam discharge unit, and to a foam transfer pipe to collect bubbles. It characterized in that it comprises a container, a foam liquefaction member provided in the interior of the collection container for liquefying bubbles, and an air outlet provided at one end of the collection container and for adjusting the air pressure.
  • the present invention provides a first step of culturing microorganisms and a culture solution in a culture tank injected with air, a second step of sensing bubbles generated in the culture tank with a foam sensor, and bubbles generated in the culture tank. It provides a method for producing a bio-surfactant comprising a third step of collecting and recovering the culture solution, and a fourth step of obtaining the bio-surfactant from the collected foam.
  • the first step is characterized in that during culturing, air in an amount of 2.5 to 10 VVM is continuously injected into the first air inlet tube.
  • the air introduced into the first air injection pipe is adjusted to an amount of 0.5 to 2.0 VVM and is introduced at intervals of 1 to 10 minutes. do.
  • the second air injection pipe is operated to inject air, and the air injection is performed in the amount of 0.5 to 1.5 VVM air per bubble sensing first time. Is injected, air is injected in the amount of 2.0 to 10 VVM in the second bubble sensing, and air is not injected in the third bubble sensing.
  • the third step of collecting the generated bubbles is step 3-1 of moving the generated bubbles to the foam discharge unit, and the bubbles moved to the foam discharge unit are moved to the foam transfer pipe. It characterized in that it comprises a step 3-2, and a third step of recovering the culture medium moved to the bubble discharge unit to the culture tank.
  • the fourth step of obtaining the bio-surfactant is characterized in that the bubble collected in the foam collection container is liquefied to obtain the bio-surfactant.
  • the bio-surfactant of the present invention is characterized in that it further comprises the step of additionally supplying a culture solution in the same amount as the culture solution in which the bubbles are liquefied after removing the bubbles in the fourth step.
  • the bio-surfactant production apparatus includes a foam sensing unit, first and second air injection pipes, and controls the operation of the first air injection pipe according to the sensing of the foam sensing unit, thereby controlling the amount of foam generated during microbial culture. It can be adjusted, and by controlling the operation of the second air inlet pipe according to the sensing of the foam sensing unit, it is possible to control the discharge of unconcentrated bubbles to obtain a high-concentrated foam, and the foam generated during microbial culture by having a foam discharge unit Loss of the culture medium can be reduced by preventing the overflow or the culture medium from being discharged with bubbles.
  • the bio-surfactant secreted in the culture solution especially surfactin
  • the bio-surfactant secreted in the culture solution is removed by collecting bubbles generated during cultivation, thereby reducing autotoxicity, thereby preventing the culture bacteria. Can protect.
  • FIG. 1 is a view showing an apparatus for producing a bio-surfactant according to an embodiment of the present invention.
  • FIG. 2 is a diagram showing an apparatus for producing a bio-surfactant according to another embodiment of the present invention.
  • FIG. 3 is a diagram showing a bubble discharge part in the bio-surfactant production apparatus according to an embodiment of the present invention, when the diameter of the upper end of the bubble discharge part is a, and the diameter of the lower end of the bubble discharge part into which the bubble flows is b, a Indicates that the diameter of is wider than the diameter of b.
  • the bio-surfactant production apparatus 10 of the present invention is a device for producing a bio-surfactant, particularly surfactin, by culturing a microorganism that produces a surfactant, and can produce a bio-surfactant economically and with improved efficiency.
  • the bio-surfactant production apparatus 10 includes an air injection unit 100 including a first air injection pipe 107 and a second air injection pipe 109, and a culture unit. (200), consisting of a bubble sensing unit 204, a bubble discharge unit 300 and a bubble collection unit 400, the bubble sensing unit 204 is the first air injection pipe 107 and the 2 Controls the operation of the air injection pipe 109.
  • an air injection unit 100 including a first air injection pipe 107 and a second air injection pipe 109
  • a culture unit. (200) consisting of a bubble sensing unit 204, a bubble discharge unit 300 and a bubble collection unit 400
  • the bubble sensing unit 204 is the first air injection pipe 107 and the 2 Controls the operation of the air injection pipe 109.
  • the culture unit 200 is for culturing microorganisms, and accommodates and cultures a culture solution containing a microorganism for producing a bio-surfactant, particularly Bacillus Bacillus for producing surfactin.
  • a bio-surfactant particularly Bacillus Bacillus for producing surfactin.
  • Microorganisms for the production of the bio-surfactants Bacillus Belle Zen sheath (B. velezensis), Bacillus amyl kwipe sieon Lowry's (B.amyloliquefaciens), Bacillus piece nipo miss (B. licheniformis), Bacillus cap Ben sheath (B. mojavensis), Bacillus pumi Ruth (B. pumilus may be), and Bacillus subtilis at least one microorganism selected from Bacillus subtilis consisting of (B.subtilis).
  • the culture unit 200 includes a culture tank 201 for culturing microorganisms, a propeller 202 for agitation of a culture solution provided at the bottom of the inside of the culture tank and for agitation of the culture solution, a low density foam provided at the top of the inside of the culture tank It is configured to include a propeller 203 for removing bubbles for extinguishing (non-concentrated bubbles), and a control sensor 206 provided inside the culture tank and for controlling the state of the culture solution.
  • a propeller 202 for stirring the culture medium a propeller 203 for removing bubbles, and a control sensor 206 are provided inside the culture tank 201.
  • the culture solution supplied to the culture tank 201 is preferably supplied at 30 to 70% of the capacity of the culture tank 201, and when cultured by supplying 70% or more of the culture solution, the culture solution overflows and the effect of collecting bubbles decreases. .
  • the propeller 202 for stirring the culture solution to agitate the culture solution the cells are prevented from sinking in the culture solution, and the air injected by the first air injection tube 107 of the air injection unit 100 described later is transferred to the culture solution. By dissolving it increases the efficiency of the culture.
  • the foam discharge unit 300 by activating the foam removal propeller 203 and blowing out the unconcentrated foam, that is, the low-density foam, together with the air injection by the second air injection pipe 109 to be described later, the foam discharge unit 300 ) To control the movement.
  • the temperature sensor 206a or the pH sensor 206b is operated as the control sensor 206 to measure the temperature and pH of the culture solution.
  • the amount of dissolved oxygen in the culture medium may be measured and the amount of foam generated may be adjusted.
  • a temperature sensor 206a or a pH sensor 206b is provided as the control sensor 206 provided in the culture tank 201, but Any control sensor installed to control the culture medium in the microbial culture apparatus in the industry may be provided, preferably a pressure sensor, a water level sensor, or a foam sensor, but is not limited thereto.
  • a foam sensing unit 204 for sensing bubbles generated from the culture unit is provided at an upper end of the culture unit 200.
  • the foam sensing unit 204 controls the operation of the first air injection pipe 107 and the second air injection pipe 109 to be described later through foam sensing, and continuously according to microbial culture in the culture tank 201
  • the generated bubbles are sensed by the bubble sensing unit 204 to operate or block the first and second air injection pipes, or to control the amount of incoming air.
  • the foam sensing unit 204 is a contact sensor that senses foam by contacting the foam generated from the surface of the culture medium of the culture tank 201 and moved to the top of the culture tank, or a laser sensor that senses the foam by recognizing the generated foam.
  • it is not limited thereto.
  • the sensor When the sensor is a contact sensor, it is preferable to be located at the top of the inside of the culture tank because the foam must directly contact the sensor, and only one contact sensor may be provided, but a plurality of contact sensors may be provided for accurate foam sensing. If the sensor is a laser sensor, it should be recognized that the bubbles generated on the surface of the culture medium move to the top of the culture tank, so it is preferable to be located on the upper side of the outside of the culture tank, and only one laser sensor is required, but accurate foam sensing is performed. For this purpose, two or more laser sensors may be provided to be located on both sides of the upper end of the culture tank.
  • a contact sensor is provided as the foam sensing unit 204 provided in the culture tank 201, but in addition to sensing materials such as foam in the art Any sensor for this can be provided.
  • the air injection unit 100 including the first air injection pipe 107 and the second air injection pipe 109 is for injecting air into the culture unit 200 or the bubble discharge unit 300, It is connected to the culture unit 200 or the discharge unit 300 to continuously or periodically inject air.
  • the first air injection pipe 107 of the air injection unit 100 is for injecting the air introduced from the air injection unit 100 into the culture solution in the culture unit 200 to the culture unit, and the bubble sensing unit 204 Until the bubble is sensed, air of 2.5 to 10 VVM is continuously introduced through the first air injection pipe 107, and after the bubble is sensed once in the bubble sensing unit 204, air of 0.5 to 2.0 VVM is Inflow and blocking are repeated at intervals of 1 to 10 minutes through the first air injection pipe 107.
  • the production amount of bio-surfactant may be increased compared to the case of continuously injecting air.
  • One or more second air injection pipes 109 of the air injection unit 100 are connected to one end of the upper end of the culture tank 201 or connected to one end of the foam discharge unit 300 to provide a foam discharge unit 300
  • the second air injection pipe 109 is formed inside, and the second air injection pipe 109 is generated from the culture unit 200 by spraying the air introduced from the air injection unit 100 from the top to the bottom of the culture tank (toward the culture solution). This is for high density by removing low-density bubbles (non-concentrated bubbles), and when the bubbles are sensed by the bubble sensing unit 204, the second air injection pipe is operated or blocked. In addition, low-density bubbles are first selectively blown out by the air injected from the second air injection pipe.
  • the second air injection pipe is activated (on) and air is introduced in an amount of 0.5 to 1.5 VVM, and when the bubble is sensed the second time, the second air is injected.
  • the amount of air introduced into the tube is increased to 2 to 10 VVM, and when the bubbles are sensed 3 times, the second air injection tube is cut off.
  • the operation control of the second air injection pipe 109 by the foam sensing unit 204 is for collecting only the concentrated foam (high-density foam) by the foam collection unit 400, which is sensed at the first and second times.
  • Low-density bubbles are first selectively extinguished by spraying air flowing into the second air injection pipe 109 by blowing unconcentrated bubbles (low-density bubbles), and the concentrated bubbles (high-density bubbles) sensed in the third time are second air.
  • the injection pipe 109 By blocking (off) the injection pipe 109, it is moved to the foam discharge unit 300 and collected by the foam collection unit 400. Since the second air injection pipe 109 needs to blow out the generated bubbles, it is preferable that air is sprayed over the largest area, and for this purpose, the second air injection pipe 109 may be in the form of a spray 111.
  • the second air injection pipe 109 has been exemplified that one second air injection pipe 109 is provided at one end of the foam discharge unit 300.
  • the second air injection pipe 109 may be provided at any position capable of extinguishing low-density bubbles generated from the culture unit 200 using a spray-type second air nozzle, and at least one second air injection A tube 109 may be provided.
  • the air injection unit 100 includes an air pump 101, an air movement pipe 105, a first valve 108 and a second valve 110.
  • the air pump 101 is equipped with a pressure gauge, is connected to the air pump 101, the air fastened to be connected to the inside of the culture tank 201 and the bubble discharge unit 300 on the other side connected to the air pump 101 Air is moved and injected into the culture tank 201 and the bubble discharge unit 300 through the moving pipe 105, in particular, the first air injection pipe 107 and the second air injection pipe 109.
  • the first valve 108 and the second valve 110 connected to the other side of the air moving pipe connected to the air pump 101 move to the first air injection pipe 107 and the second air injection pipe 109, respectively. And controlling incoming air.
  • the air injection unit 100 a first air flow meter 102 for adjusting the flow of air injected from the air pump 101, an air filter 103 for filtering the supplied air, A water tank 104 for supplying moisture to the air, and a second air flow meter 106 for measuring the injected air flow may be additionally provided in the air injection unit 100 as necessary, and the above member By providing it, it is possible to control the flow of air.
  • the foam discharge unit 300 is for collecting the concentrated foam discharged to the foam discharge unit inlet 301 and recovering the culture solution to the culture unit 200, the foam discharge port 205 of the culture tank 201 ) And the generated bubbles are continuously or intermittently collected, and the culture solution that has moved to the bubble discharge unit 300 along with the bubbles is recovered to the culture tank 201.
  • the foam discharge unit 300 is connected to the upper portion of the culture tank 201 and is concentrated by the foam sensing unit 204 and the first and second air injection pipes 107 and 109 to the culture tank 201
  • the concentrated foam (high-density foam) that has moved from the lower part of the to the upper part is collected, and some of the culture liquid that has moved together with the foam is recovered to the culture tank 201 again.
  • the high-density foam (concentrated foam) collected in the foam discharge unit 300 is collected into the foam collection container 402 through a foam transfer pipe 401 connected to the upper portion of the foam discharge unit 300 to be described later.
  • the foam discharge unit 300 and the foam transfer pipe 401 are vertically connected to the top of the culture tank 201 to prevent the phenomenon that some of the culture liquid moved along the foam moves to the foam collection container 402 due to gravity. I can.
  • the bubble discharge part 300 is connected to the bubble discharge part inlet 301 of the bubble discharge part and the bubble discharge port 205 of the culture tank 201. In addition, as shown in FIGS.
  • the bubble discharge unit 300 may be configured in a bottleneck shape, and the diameter of the upper end of the bubble discharge unit 300 is greater than the diameter of the lower end of the bubble discharge unit 300 into which the bubbles are introduced. It may be formed to be wide. Specifically, when the top diameter of the bubble discharge part 300 is a, and the diameter of the lower end of the bubble discharge part 300 into which the bubble flows is b, the diameter of a may be larger than the diameter of b, and the The diameter of a may be 2 to 10 times wider than the diameter of b.
  • the bubble collection unit 400 is for collecting and collecting the bubbles generated in the culture tank 201 by the bubble discharge unit 300, and continuously or intermittently collects the generated bubbles.
  • the foam collection unit 400 includes a foam moving pipe 401, a foam collection container 402, a foam liquefaction member 403, and an air outlet 404.
  • the bubble collection container 402 is provided with a bubble liquefaction member 403 for liquefying bubbles, and a propeller 403a for liquefying bubbles and/or a blower 403b to liquefy the collected bubbles to thereby liquefy the collected bubbles and biointerface
  • An active agent, in particular surfactin, is obtained.
  • a foam liquefaction propeller 403a and/or a blowing device 403b as a foam liquefaction member 403 provided in the foam collection container 402
  • the collected foam may be transferred to another container and allowed to liquefy by leaving it in a refrigerator for a certain period of time.
  • an air outlet 404 for adjusting air pressure is provided at one end of the collection container 402.
  • bio-surfactant production apparatus 10 By using the bio-surfactant production apparatus 10 as described above, the production efficiency of a bio-surfactant, particularly surfactin, can be improved, and a specific method is as follows.
  • the method for producing a bio-surfactant includes a first step of culturing microorganisms and a culture medium in a culture tank injected with air, a second step of sensing bubbles generated in the culture tank with a foam sensor, and collecting bubbles generated in the culture tank. And, it can be divided into a third step of recovering the culture solution, and a fourth step of obtaining a bio-surfactant from the collected foam.
  • the first step is a step of culturing by supplying a microorganism producing a bio-surfactant and a culture solution to the inside of the culture tank 201 and injecting air.
  • the culture solution supplied to the culture tank 201 is 30 compared to the capacity of the culture tank 201 It is preferable to supply in an amount of 70% to 70%, and when the culture solution is cultured by supplying 70% or more of the culture solution, the culture solution overflows and the effect of collecting bubbles decreases.
  • microorganisms that produce bio-surfactants, especially surfactin are Bacillus velezensis , Bacillus amyloliquefaciens, Bacillus licheniformis , and Bacillus mobbenscis . (B.
  • Bacillus pumi loose may be a (B. pumilus), and Bacillus subtilis Bacillus subtilis one or more kinds selected from the group consisting of (B.subtilis), not limited to this.
  • Bacillus subtilis Bacillus subtilis one or more kinds selected from the group consisting of (B.subtilis), not limited to this.
  • bubbles may be generated by supplying air, and a specific method of generating bubbles will be described in detail below.
  • the method of generating bubbles by culturing the culture solution injected with air is to generate bubbles by injecting air according to the operation of the air pump 101 and activating the propeller 202 for stirring the culture solution provided in the culture tank 201.
  • air injection according to the operation of the air pump 101 air in an amount of 2.5 to 10 VVM is continuously introduced into the culture tank through the first air injection pipe 107.
  • the time for generating the foam is not particularly limited, but foam may be repeatedly generated for a short time, such as 20 to 30 minutes, or may be continuously generated for 6 hours or more.
  • the second step is a step of sensing the foam generated in the culture tank with a foam sensor, which is the most essential step for improving the production efficiency of a bio-surfactant, particularly surfactin, in the present invention.
  • the initial foam generated during cultivation is an unconcentrated foam (low density foam), and since the amount of the bio-surfactant contained in the initial foam is very small, it is difficult to collect the low-density foam to obtain a bio-surfactant in high yield. Accordingly, as described later, a method for collecting only the concentrated foam was devised.
  • the operation of the first and second air injection pipes 107 and 109 is controlled. Specifically, when the bubble is sensed, the amount of air continuously flowing into the first air injection pipe 107 is adjusted to 0.5 to 2.0 VVM in the first step and periodically flows in, preferably 0.5 to 2.0 VVM amount of air. Is injected and blocked at intervals of 1 to 10 minutes. Such control of the first air injection pipe 107 may increase the production amount of bio-surfactant.
  • the second air injection pipe 109 is operated to inject or block air into the culture tank 201.
  • air in an amount of 0.5 to 1.5 VVM is injected in the first time of bubble sensing, and the second time of bubble sensing is injected by increasing the amount of air in an amount of 2.0 to 10 VVM, and the third time of bubble sensing From then on, air injection is stopped.
  • Control of the second air inlet pipe 109 as described above prevents unconcentrated bubbles (low-density bubbles) from being discharged to the bubble discharge unit 300 and collected by the bubble collection unit 400 and concentrates the bubbles. It can increase the production of surfactants.
  • the third step is a step of collecting the generated bubbles in the bubble collection container 402, which includes the step of collecting the concentrated bubbles (high density bubbles) by moving the generated bubbles to the bubble discharge unit 300, , This step is also essential in order to obtain a bio-surfactant in high yield.
  • surfactin a bio-surfactant having an antibiotic effect
  • Bacillus bacteria do not grow further and die because of the surfactin produced by themselves, so there is a difficulty in increasing the production of surfactin.
  • cells are essential for the production of surfactin, and the number of cells must be increased to increase the production of surfactin, but if the number of cells is too large, it results in disturbing the production of surfactin through quorum sensing.
  • the concentration of surfactin is lowered during culturing so that host cells are not killed by surfactin, or the number of cells is appropriately reduced or the cells do not grow further during culturing. It was determined that the production of cotton surfactin could be increased, and a method of collecting foam was devised as described later.
  • the third step of collecting the generated bubbles is a step 3-1 of moving the generated bubbles to the bubble discharge unit 300 through the bubble discharge port 205, to the bubble discharge unit 300 Bubbles generated through the 3-2 step of moving the moved bubbles to the bubble transfer pipe 401 and the 3-3 step of recovering the culture solution moved to the bubble discharge unit 300 to the culture tank 201 It is collected in the collection container 402.
  • the collection container 402. In general, when bubbles are collected without the foam discharge unit 300, some of the bubbles overflow, and when the bubbles are recovered, some culture solutions are also recovered, so that the culture solution and the produced bio-surfactant are lost.
  • bio-surfactant is low, and bio-surfactant-producing bacteria do not survive due to the bio-surfactant produced at high concentration in the culture medium, making it difficult to continuously produce bio-surfactant.
  • bubbles are collected through the bubble discharge unit 300 as in the present invention, some cells present in the collected bubbles are removed to delay the formation of durable bodies through quorum sensing, thereby prolonging the period for producing surfactants.
  • it is possible to continuously produce bio-surfactants, and because a high-concentration bio-surfactant is present in the collected foam, the concentration of the bio-surfactant produced in the culture medium can be lowered, thereby forming conditions in which the bio-surfactant can continue to grow. , It can improve the production efficiency of bio-surfactants.
  • the fourth step is a step of collecting the bio-surfactant, and through the third step, the bubble collected in the bubble collection container 402 may be liquefied to obtain a bio-surfactant.
  • the liquefaction of the foam is generally any method capable of liquefying the foam, but preferably, a foam liquefaction propeller (403a) or a blower (403b) installed inside the foam collection container 402 of the present invention is used. To liquefy it, or if the bubble liquefaction member is not installed in the bubble collection container 402, the bubble may be collected and allowed to stand in a refrigerating chamber to liquefy.
  • the bio-surfactant generating device of the present invention and the foam sensing unit and the second air-injection pipe are both provided. It was confirmed that surfactin was generated by a bio-surfactant generating device without an air injection tube. The results are shown in Table 2 below. Indicated.
  • Bubble sensing unit 2nd air inlet pipe Surfactin production Main invention ⁇ ⁇ 2.2g Comparative Example 3 ⁇ ⁇ 0.9 ⁇ 1.6g
  • the bio-surfactant generating device of the present invention equipped with a foam sensing unit and a second air inlet tube generates concentrated foam (high-density foam) to increase the production of surfactin, whereas the foam sensing unit And in the apparatus of Comparative Example not provided with the second air inlet tube, the foam was not concentrated, so the production amount of surfactin decreased.
  • the amount of air injected through the first air injection pipe and the amount of surfactin produced are inversely proportional.
  • the second air inlet pipe was used to confirm the surfactin generation under various air inlet conditions.
  • the air inflow conditions and the surfactin production amount of the second air injection pipe are shown in Table 3 below.
  • Bubble sensing unit 2nd air inlet pipe First round air volume Secondary air volume Surfactin production Main invention ⁇ ⁇ 1.0 VVM 5.0 VVM 2.2g Comparative Example 4 ⁇ ⁇ 1.0 VVM 1.0 VVM 0.9g
  • the concentration of the foam was calculated by comparing the amount of the foam produced once after culturing for 60 hours and the amount of cells and surfactin in the remaining culture solution, and the results are shown in Table 4 below.

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Abstract

La présente invention concerne un appareil de production de biotensioactif comprenant un premier et un second tuyau d'injection d'air pour concentrer la mousse et récupérer un milieu de culture, une partie de détection de mousse, et une partie d'évacuation de mousse. L'invention concerne une partie de détection de mousse et un premier et un second tuyau d'injection d'air, l'appareil de production de biotensioactif peut obtenir une mousse hautement concentrée en régulant une quantité et une concentration de mousse et peut réduire la perte du milieu de culture en empêchant le débordement de la mousse générée pendant la culture de micro-organismes ou la décharge du milieu de culture conjointement avec la mousse. En outre, lorsqu'elle est cultivée à l'aide de l'appareil de production de biotensioactif selon la présente invention, le micro-organisme peut être protégé. Certaines cellules présentes dans la mousse collectée sont éliminées pour prolonger la durée de production d'un tensioactif, ce qui permet d'améliorer le rendement de production du biotensioactif.
PCT/KR2020/009785 2019-07-25 2020-07-24 Appareil de production de biotensioactif et procédé de production de biotensioactif l'utilisant WO2021015587A1 (fr)

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KR1020190090419A KR102240151B1 (ko) 2019-07-25 2019-07-25 바이오 계면활성제의 생산 장치 및 이를 이용한 바이오 계면활성제의 생산 방법
KR10-2019-0090419 2019-07-25
KR10-2020-0083635 2020-07-07
KR1020200083635A KR102432456B1 (ko) 2020-07-07 2020-07-07 바이오 계면활성제의 생산 장치 및 이를 이용한 바이오 계면활성제의 생산 방법

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Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4883759A (en) * 1983-07-12 1989-11-28 Phillips Petroleum Company Fermentation method and apparatus
US5476573A (en) * 1991-11-22 1995-12-19 Ajinomoto Co., Inc. Apparatus for defoaming and controlling aerobic culture fermentation
KR100737709B1 (ko) * 1999-08-13 2007-07-11 다우 코닝 에스. 아. 실리콘 거품 조절제
KR20140019406A (ko) * 2011-03-29 2014-02-14 다니스코 유에스 인크. 거품 제어 방법
CN208378874U (zh) * 2018-05-21 2019-01-15 南京工业大学 一种循环消泡系统
KR101989348B1 (ko) * 2018-03-02 2019-06-20 한국에너지기술연구원 생물반응기 내부의 폼을 제거하는 트랩 및 이를 사용하여 미생물 배양시 발생하는 폼을 제거하는 방법

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4883759A (en) * 1983-07-12 1989-11-28 Phillips Petroleum Company Fermentation method and apparatus
US5476573A (en) * 1991-11-22 1995-12-19 Ajinomoto Co., Inc. Apparatus for defoaming and controlling aerobic culture fermentation
KR100737709B1 (ko) * 1999-08-13 2007-07-11 다우 코닝 에스. 아. 실리콘 거품 조절제
KR20140019406A (ko) * 2011-03-29 2014-02-14 다니스코 유에스 인크. 거품 제어 방법
KR101989348B1 (ko) * 2018-03-02 2019-06-20 한국에너지기술연구원 생물반응기 내부의 폼을 제거하는 트랩 및 이를 사용하여 미생물 배양시 발생하는 폼을 제거하는 방법
CN208378874U (zh) * 2018-05-21 2019-01-15 南京工业大学 一种循环消泡系统

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