CN219907680U - Random sweep frequency ultrasonic wave industrial bioreactor - Google Patents
Random sweep frequency ultrasonic wave industrial bioreactor Download PDFInfo
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- CN219907680U CN219907680U CN202223352479.4U CN202223352479U CN219907680U CN 219907680 U CN219907680 U CN 219907680U CN 202223352479 U CN202223352479 U CN 202223352479U CN 219907680 U CN219907680 U CN 219907680U
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- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The utility model discloses a random sweep frequency ultrasonic industrialized bioreactor, and belongs to the field of biological manufacturing. The utility model relates to a random sweep frequency ultrasonic industrialized bioreactor which consists of a tank body, a stirrer, an ultrasonic generator, an ultrasonic transducer, an ultrasonic mixed flow baffle, a control cabinet, a PLC touch screen and the like. The ultrasonic generator has three working modes of fixed frequency, periodic frequency sweep and random frequency sweep, wherein the random frequency sweep working mode eliminates resonance effect and damage to microorganism cells. The ultrasonic mixed flow baffle combines the ultrasonic vibration plate and the traditional mixed flow baffle into one component, thereby solving the problem of long-distance transmission of ultrasonic waves in a large-size industrial bioreactor. The device can be applied to not only microbial fermentation, but also strengthening of liquid processing processes such as chemical reaction, enzyme activation, emulsification modification, separation extraction and the like.
Description
Technical Field
The utility model relates to a random sweep ultrasonic industrialized bioreactor and application thereof, belonging to the field of biological manufacturing.
Background
In recent years, in order to improve the microbial fermentation efficiency, more and more scholars are investigating the application of ultrasonic waves to the promotion of the fermentation process. However, the current research work and solutions suffer from two disadvantages. Firstly, the grasping of the sound field intensity is very difficult, the activating effect of the microorganism is not obvious due to the fact that the sound intensity is too low, and the activity of the microorganism can be inhibited due to the fact that the sound intensity is too high; secondly, the large size of the industrial bioreactor brings great difficulty to the long-distance effective transmission of ultrasonic waves.
For the problem of sound field intensity regulation, the current problem is mainly realized by changing power and frequency. The adjustment of the ultrasonic power is simpler and the adjustment of the ultrasonic frequency is more complex. Besides the traditional single-frequency continuous working modes, the ultrasonic wave also comprises pulse ultrasonic wave and multi-frequency ultrasonic wave, and the multi-frequency ultrasonic wave is divided into two working modes of sequential ultrasonic wave and synchronous ultrasonic wave. In order to expand the spectrum of ultrasonic waves, the inventor of the present patent invented a swept-frequency ultrasonic device in 2009, and the device emits a periodic swept-frequency ultrasonic (for short, periodic swept-frequency ultrasonic) (ZL 200910212707.8). The sweep frequency ultrasonic wave means that the frequency of the ultrasonic wave fluctuates up and down around the center frequency within a small amplitude range according to a certain rule along with the extension of the working time. According to the principle of acoustics, a strong vibration force is generated when the frequency of ultrasonic waves passes through the center frequency, so that the strong vibration force of the sweep ultrasonic waves has better activating effect on microorganisms under the same power. However, the periodic vibration force generated by the periodic sweep ultrasonic wave can further form a stronger resonance effect, and the resonance effect is easy to cause serious damage to the microbial cells, so that the activity of the microbial cells is reduced. Therefore, there is a need for further improvements in swept frequency ultrasound.
Regarding the problem of large size of industrial bioreactors, at present, ultrasonic waves are applied to bioreactors, and there are schemes of inserting an ultrasonic probe into a tank body center (ZL 201720365631.2), immersing an ultrasonic vibration plate into a tank bottom (ZL 201620687544.4) and the like, wherein a mechanical stirrer cannot be installed at the center of the tank body, and the ultrasonic wave transmission distance of the immersed tank bottom is limited, so that the requirements of the large-size industrial bioreactors cannot be met. Therefore, it is necessary to design a mechanism that can more satisfy the addition of ultrasonic waves to large-sized industrial bioreactors.
Disclosure of Invention
The utility model aims to provide a random sweep ultrasonic industrialized bioreactor, which designs a random sweep ultrasonic working mode (called random sweep ultrasonic for short) on the premise of keeping the advantages of sweep ultrasonic, and the time of the ultrasonic frequency passing through the center frequency is not periodic and is random, so that the vibration force generated by the ultrasonic at the center frequency can not form a resonance effect, thus the microbial cells can not be damaged, and the aim of remarkably activating the microbial activity by utilizing the sweep ultrasonic is effectively realized.
In order to solve the problem of effective ultrasonic transmission in large-size industrial bioreactors, the utility model designs the mixed flow baffle vertically arranged in the tank wall into a hollow baffle with an ultrasonic transducer. The fermentation liquor rotates in the bioreactor under the pushing of the stirrer and continuously impacts the mixed flow baffle plate to receive the action of ultrasonic waves. Therefore, the baffle plate with the built-in ultrasonic transducer plays a role in mixing flow of the traditional baffle plate, plays a role in promoting the growth of microorganisms by ultrasonic waves on the other hand, can conveniently transfer ultrasonic energy from the periphery of the inner wall of the bioreactor to the center, and overcomes the difficulty of long-distance ultrasonic wave transfer in large-size industrial bioreactors.
The utility model aims at realizing the following technical scheme:
the utility model relates to a random sweep frequency ultrasonic industrialized bioreactor, which consists of a tank body, a stirrer, an ultrasonic transducer, an ultrasonic mixed flow baffle, a control cabinet, a PLC touch screen and an ultrasonic generator, wherein the stirrer is vertically arranged at the central axis of the tank body; the ultrasonic transducer is connected with the ultrasonic generator through a cable, and the sensor interface is arranged on the tank wall of the lower half part of the tank body.
The ultrasonic generator has three working modes of fixed frequency, periodic frequency sweep and random frequency sweep, and can be set through a PLC touch screen on a control cabinet. The frequency of the ultrasonic wave can be single frequency or multi-frequency; the multiple frequencies can work synchronously or sequentially. The frequency of the ultrasonic wave is between 20kHz and 80kHz, and the power density is between 20W/L and 160W/L; the sweep frequency amplitude is between 0 and +/-2 kHz.
The number of the ultrasonic mixed flow baffles is 4-10.
The random sweep ultrasonic industrialized bioreactor can be applied to efficient fermentation of various microorganisms. After the raw material liquid is pumped into the bioreactor, microorganism strains are inoculated, parameters such as fermentation temperature, rotation speed of a stirrer, ultrasonic working mode, power density, sweep frequency amplitude, ultrasonic starting time and working time, fermentation time and the like are set on a PLC touch screen of a control cabinet, the stirrer is started, heating circulating water is started for heating, pH value is regulated, and fermentation is started.
The bioreactor can be applied to microbial fermentation and strengthening of other liquid processing processes such as chemical reaction, enzyme activation, emulsification modification, separation and extraction and the like.
Drawings
FIG. 1 is a schematic diagram of a random swept ultrasonic industrial bioreactor, wherein: 1. a tank body; 2. a stirrer; 3. a sensor interface; 4. a cable; 5. an ultrasonic transducer; 6. an ultrasonic mixed flow baffle; 7. a control cabinet; 8. a PLC touch screen; 9. an ultrasonic generator;
fig. 2 is a schematic view of an ultrasonic mixed flow baffle structure, wherein: 4. a cable; 5. an ultrasonic transducer; 6. an ultrasonic mixed flow baffle;
fig. 3 is a schematic diagram of the ultrasonic frequency variation of the fixed frequency (a), the periodic sweep (b) and the random sweep (c).
The utility model has the advantages that:
(1) The adoption of random sweep frequency ultrasonic wave to strengthen the fermentation of microorganisms not only maintains the advantage of strong effect of the sweep frequency ultrasonic wave, but also eliminates the occurrence of resonance effect and avoids the damage of the resonance effect to microorganism cells.
(2) The ultrasonic vibration plate and the mixed flow baffle are combined into one component, so that the problem of long-distance transmission of ultrasonic waves in a large-size industrial bioreactor is solved, and the ultrasonic vibration plate and the mixed flow baffle are suitable for industrial amplification application of technology.
Detailed Description
Example 1:
the utility model discloses a random sweep frequency ultrasonic industrialized bioreactor which is shown in figure 1 and consists of a tank body 1, a stirrer 2, a sensor interface 3, a cable 4, an ultrasonic transducer 5, an ultrasonic mixed flow baffle 6, a control cabinet 7, a PLC touch screen 8, an ultrasonic generator 9 and the like. The stirrer 2 is vertically arranged at the central axis of the tank body 1, the sensor interface 3 is arranged on the tank wall of the lower half part of the tank body 1, and the ultrasonic mixed flow baffles 6 are uniformly distributed and vertically arranged on the inner wall of the tank body. The structure of the ultrasonic mixed flow baffle is shown in figure 2, the ultrasonic transducer 5 is stuck in the mixed flow baffle 6, the ultrasonic generator 9 is arranged in the control cabinet 7, and the ultrasonic transducer 5 is connected with the ultrasonic generator 9 through the cable 4.
The ultrasonic generator has three working modes of fixed frequency, periodic frequency sweep and random frequency sweep (see figure 3), and can be set through a PLC touch screen on a control cabinet. The frequency of the ultrasonic wave can be single frequency or multi-frequency; the multiple frequencies can work synchronously or sequentially. The frequency of the ultrasonic wave is between 20kHz and 80kHz, and the power density is between 20W/L and 160W/L; the sweep frequency amplitude is between 0 and +/-2 kHz.
Example 2: liquid fermentation of bacillus subtilis
The strain is bacillus subtilis Bacillus subtilis 10160 and is purchased from China industry microbiological culture collection center. This is an example of the use of the random swept ultrasonic industrial bioreactor of the present utility model to increase biomass, soluble protein production and polypeptide production during the fermentation of bacillus subtilis. The fermentation medium consists of: 20% of soybean meal, 2% of corn gluten meal and 1% of KH 2 PO 4 The inoculation amount is 10%. The fermentation conditions are as follows: the fermentation temperature is 36 ℃, the ultrasonic power density is 80W/L, the ultrasonic frequency is 33+/-1 kHz, and the disposable ultrasonic treatment is carried out for 1h on the 3 rd day after inoculation. Under these conditions, the biomass was 4.67×10 9 The cfu/mL is respectively increased by 146.78 percent, 62.89 percent and 31.65 percent compared with a control group without ultrasonic, fixed frequency ultrasonic (33 kHz) and periodic sweep frequency ultrasonic (33+/-1 kHz); the content of soluble protein in the fermentation product is 22.72mg/mL, which is respectively improved by 17.21%, 11.01% and 7.91% compared with a control group without ultrasound, fixed frequency ultrasound and periodic sweep frequency ultrasound; the polypeptide content of the fermentation product is 22.72mg/mL, which is respectively increased by 27.28, 18.21 percent and 14.19 percent compared with the control group without ultrasonic, fixed-frequency ultrasonic and periodic sweep frequency ultrasonic.
Example 3: phellinus liquid fermentation
The strain is Phellinus (preservation number is CGMCC No.17078, china center for Industrial microorganism culture Collection). The method is an embodiment for improving the synthesis yield of mycelium flavonoids and polysaccharides in the fermentation process of Phellinus by using the random sweep ultrasonic industrialized bioreactor. The components of the fermentation medium are as follows (g/L): flour (50), bran (15), mulberry twig powder (10), KH 2 PO 4 (2)，MgSO 4 (1). The inoculation amount is 8%. The fermentation conditions are as follows: the inoculation amount of the strain is 10%, the fermentation temperature is 26 ℃, the fermentation time is 10 days, ultrasonic waves are applied on the 4 th to 8 th days of fermentation, the ultrasonic waves are applied for 1 time each day for 10 minutes, the ultrasonic power density is 120W/L, and the ultrasonic frequency is 40+/-2 kHz. Under the condition, the mycelium flavone yields are 55.81mg/L respectively, which are higher than those of non-ultrasonic, fixed-frequency ultrasonic (40 kHz) and periodic sweep ultrasonic (40+ -2 k)Hz) control group increased by 68.82%, 46.23%, 38.17%, respectively; the mycelium polysaccharide yields are 2912.20mg/L respectively, which are improved by 24.06%, 19.21% and 16.25% respectively compared with the control group without ultrasound, fixed frequency ultrasound and periodic sweep frequency ultrasound.
Example 4: bacillus amyloliquefaciens liquid fermentation
The strain is bacillus amyloliquefaciens (CICC 10160) purchased from China industry microbiological culture collection center. The method is an embodiment of strengthening liquid fermentation seed production by adopting the random sweep ultrasonic industrialized bioreactor before solid state fermentation, and finally aims to improve the protease yield, the soluble protein content and the polypeptide content of a fermentation product in solid state fermentation of the oil cake. Liquid fermentation: inoculating mother liquor into 50L nutrient broth culture medium according to inoculum size of 1% (mL/mL), fermenting at 37deg.C for 10 hr, and starting ultrasonic generator to continue fermentation for 20min. The ultrasonic treatment adopts a double-frequency synchronous random sweep frequency working mode, the frequency combination is 40+/-2/60+/-2 kHz, and the power density is 40W/L. Solid state fermentation: inoculating the strain liquid into soybean meal according to the amount of 100mL/kg, and supplementing the insufficient water amount with hot water at 37 ℃ according to the ratio of the total feed liquid to 1:1 (g/mL). After all is ready, solid state fermentation is started. The polypeptide content of the ultrasonic treatment group is 153.68g/kg in the fermentation 48, which is improved by 11.91 percent, 4.21 percent and 2.29 percent respectively compared with the control group without ultrasonic, fixed frequency ultrasonic (40/60 kHz) and periodic sweep frequency ultrasonic (40+/-2/60+/-2 kHz), and is 7.2 times of the soybean meal raw material.
Claims (4)
1. The random sweep frequency ultrasonic industrialized bioreactor is characterized by comprising a tank body, a stirrer, an ultrasonic transducer, an ultrasonic mixed flow baffle, a control cabinet, a PLC touch screen and an ultrasonic generator, wherein the stirrer is vertically arranged at the central axis of the tank body; the ultrasonic transducer is connected with the ultrasonic generator through a cable, and the sensor interface is arranged on the tank wall of the lower half part of the tank body.
2. The ultrasonic industrialized reactor with random frequency sweep according to claim 1, wherein the ultrasonic generator has three working modes of fixed frequency, periodic frequency sweep or random frequency sweep, and the frequency of the ultrasonic wave is single frequency or multi-frequency; the multi-frequency is synchronous or sequential; the frequency of the ultrasonic wave is between 20kHz and 80kHz, and the power density is between 20W/L and 160W/L; the sweep frequency amplitude is between 0 and +/-2 kHz.
3. The ultrasonic industrial bioreactor with random frequency sweep according to claim 1, wherein the number of the ultrasonic mixed flow baffles is 4-10.
4. The random sweep ultrasonic industrialized bioreactor according to claim 1, wherein the ultrasonic mixed flow baffle combines the ultrasonic vibration plate and the traditional mixed flow baffle into one component, thereby solving the problem of long-distance transmission of ultrasonic waves in large-size industrialized bioreactors.
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| CN202223352479.4U CN219907680U (en) | 2022-12-13 | 2022-12-13 | Random sweep frequency ultrasonic wave industrial bioreactor |
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| CN202223352479.4U CN219907680U (en) | 2022-12-13 | 2022-12-13 | Random sweep frequency ultrasonic wave industrial bioreactor |
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