Directional micro-ecological bioreactor

Abstract

The utility model relates to a directional micro-ecological bioreactor. The directional micro-ecological bioreactor comprises a reactor tank body and a reactor cover body, the interior of the reactor tank body is hollow, and a containing cavity with one closed end and the other open end is formed; the reactor cover body is detachably connected to the reactor tank body and is used for sealing the open end of the accommodating cavity; a pressure valve port, a temperature sensor probe pipe port, a sampling pipe port and an aeration pipe port are formed in the covering part of the reactor cover body; a pressure valve is arranged at the pressure valve port, a temperature sensing detector is inserted into the temperature sensor probe pipe port, a sampling pipe is inserted into the sampling pipe port, and an aeration pipe is inserted into the aeration pipe port. The directional micro-ecological bioreactor is simple in structure and convenient to operate, and has fermentation stability and reliability.

Description

Technical Field

[0001] The present disclosure relates to the field of fermentation technology, and in particular, to a directional microecological bioreactor. Background Art

[0002] A bioreactor is a device used for in vitro culture of microorganisms and cells, and obtaining products through biochemical reactions or fermentation processes. It is one of the key devices in the fields of biosynthesis, biomanufacturing, and biopharmaceuticals. A bioreactor can control multiple units in the same system to maintain a high degree of consistency in various parameters. It is an important link in the key technology process design of the industrialization of synthetic biology such as microorganisms. Its core functions are high-throughput screening and process amplification. Although there are many types of bioreactors and rich models currently, these bioreactors are costly.

[0003] In the prior art, Patent CN220246127U discloses an aeration-magnetic stirring composite microalgae culture photobioreactor, which is a reactor for algae culture. It does not consider the problem of contamination by miscellaneous bacteria during the bacteria culture process, and there are problems such as the overall reactor tank cannot be sterilized uniformly and the temperature cannot be controlled constantly, resulting in low culture efficiency. In addition, the ventilation disc and the stirring element are located on both sides, and the stirring speed and the ventilation volume cannot be quantitatively controlled, seriously affecting the aeration efficiency.

[0004] Currently, commercially available bioreactors are expensive and have high usage costs. The scale-up process of laboratory bioreactors is unstable, requires repeated testing, involves a large amount of work, is complex to operate, has high requirements for personnel, and cannot meet the current market demand. Summary of the Utility Model

[0005] The purpose of the present disclosure is to provide a directional microecological bioreactor, which has a simple structure, is convenient to operate, and has fermentation stability and reliability.

[0006] To achieve the above object, the present disclosure provides a directional microecological bioreactor, which includes a reactor tank body and a reactor cover body;

[0007] The interior of the reactor tank body is hollow and forms a receiving cavity that is closed at one end and open at the other end; the reactor cover body is detachably connected to the reactor tank body for closing the open end of the receiving cavity;

[0008] A pressure valve port, a temperature sensor probe port, a sampling pipe port, and an air supply pipe port are provided on the covering part of the reactor cover body; a pressure valve is provided at the pressure valve port, a temperature sensing detector is inserted into the temperature sensor probe port, a sampling pipe is inserted into the sampling pipe port, and an air supply pipe is inserted into the air supply pipe port.

[0009] Optionally, the temperature measuring part of the temperature sensing detector is located inside the reactor tank, and the fixing part of the temperature sensing detector is detachably connected to the reactor cover at the temperature sensor probe port.

[0010] Optionally, the inlet of the sampling tube is located inside the reactor tank; the outlet of the sampling tube is located outside the reactor tank and is connected with a sample guiding tube, and a first straight-through valve is arranged on the sample guiding tube; the fixing part of the sampling tube is detachably connected to the reactor cover at the sampling tube port.

[0011] Optionally, the outlet of the air supply pipe is located inside the reactor tank; the inlet of the air supply pipe is located outside the reactor tank and is connected with an air guiding pipe, and the inlet of the air guiding pipe is connected with an air guiding pump; a bacteria filter, an air flow meter and a second straight-through valve are sequentially arranged along the direction from the inlet to the outlet of the air guiding pipe; the fixing part of the air supply pipe is detachably connected to the reactor cover at the air supply pipe port.

[0012] Optionally, the air supply pipe includes a gas introduction part and an aeration part, the aeration part is a pipeline with an annular structure, and a plurality of air holes are arranged on the aeration part.

[0013] Optionally, the air holes are circular, with a pore diameter of 0.5 - 1.0 mm; the number of the air holes is 10 - 14.

[0014] Optionally, a magnetic rotor is further installed inside the reactor tank.

[0015] Optionally, the number of the magnetic rotors is 1.

[0016] Optionally, a magnetic stirring constant temperature water bath is further included for heating the reactor tank body.

[0017] Optionally, the volume of the reactor tank body is 300 - 500 mL.

[0018] Through the above technical solutions, the present disclosure provides a directional microecological bioreactor, which has a simple structure and convenient operation, can realize the amplification of bacterial strains, and has the stability and reliability of fermentation.

[0019] Other features and advantages of the present disclosure will be described in detail in the subsequent specific implementation part. Description of the Drawings

[0020] The drawings are used to provide a further understanding of the present disclosure, and constitute a part of the specification, and are used to explain the present disclosure together with the following specific implementation manners, but do not constitute a limitation to the present disclosure. In the drawings:

[0021] Figure 1It is a schematic structural diagram of the directional microecological bioreactor of the present disclosure.

[0022] Figure 2 It is a schematic structural diagram after the reactor tank body and the reactor cover body of the present disclosure are assembled.

[0023] Figure 3 It is a schematic structural diagram of the covering part of the reactor cover body of the present disclosure.

[0024] Figure 4 It is a schematic structural diagram of the aeration pipe of the present disclosure.

[0025] Explanation of reference numerals

[0026] 1 Pressure valve port, 2 Temperature sensor probe port, 3 Sampling pipe port

[0027] 4 Aeration pipe port, 5 Magnetic rotor, 6 Reactor cover body

[0028] 7 Reactor tank body, 8 Magnetic stirring constant temperature water bath, 9 Air guide pump

[0029] 10 Bacteria filter, 11 Air flow meter, 12 Air guide pipe

[0030] 13 Second straight-through valve, 14 First straight-through valve, 15 Accommodating cavity

[0031] 16 Introduction part, 17 Aeration part, 18 Sampling guide pipe Detailed implementation manners

[0032] The following will describe the detailed implementation manners of the present disclosure with reference to the accompanying drawings. It should be understood that the detailed implementation manners described herein are only for explaining and interpreting the present disclosure, and are not used to limit the present disclosure.

[0033] As Figure 1 and 2 shown, the present disclosure provides a directional microecological bioreactor, which includes a reactor tank body 7 and a reactor cover body 6;

[0034] The interior of the reactor tank body 7 is hollow and forms an accommodating cavity 15 that is closed at one end and open at the other end; the reactor cover body 6 is detachably connected to the reactor tank body 7 and is used to close the open end of the accommodating cavity 15;

[0035] The covering part of the reactor cover body 6 is provided with a pressure valve port 1, a temperature sensor probe port 2, a sampling pipe port 3 and an aeration pipe port 4; a pressure valve is provided at the pressure valve port 1, a temperature sensing detector is inserted into the temperature sensor probe port 2, a sampling pipe is inserted into the sampling pipe port 3, and an aeration pipe is inserted into the aeration pipe port 4.

[0036] In the present disclosure, when the directional microecological bioreactor is in use, the reactor cover 6 and the reactor tank 7 are in a connected state. The present disclosure does not specifically limit the connection state between the reactor cover 6 and the reactor tank 7. For example, it can be a screw connection, as long as it can ensure the airtightness of the accommodation cavity 15 after the reactor cover 6 and the reactor tank 7 are connected. This directional microecological bioreactor has a simple structure and convenient operation, can achieve the amplification of microorganisms, and has fermentation stability and reliability.

[0037] As Figure 2 shown, where H7 represents the height of the reactor tank 7, the reactor tank 7 is cylindrical, and φ7 represents the diameter of the cylindrical reactor tank 7. As Figure 3 shown, represents the diameter of the reactor cover 6. The pressure valve port 1, the temperature sensor probe port 2, the sampling port 3, and the aeration pipe port 4 are each independently circular. Among them, represents the diameter of the pressure valve port 1, represents the diameter of the temperature sensor probe port 2, represents the diameter of the sampling port 3, represents the diameter of the aeration pipe port 4.

[0038] According to the present disclosure, when the directional microecological bioreactor is in use, the pressure valve can release the gas in the accommodation cavity 15 of the reactor tank 7 to prevent the pressure in the accommodation cavity 15 from being too high. Preferably, the diameter of the pressure valve port 1 can be 15 mm.

[0039] In one embodiment, when the directional microecological bioreactor is in use, the temperature measuring part of the temperature sensing detector is located inside the reactor tank 7 and below the liquid level of the culture medium in the reactor tank 7; the fixing part of the temperature sensing detector is detachably connected to the reactor cover 6 at the temperature sensor probe port 2. In the present disclosure, the connection method of the fixing part of the temperature sensing detector being detachably connected to the reactor cover 6 is not specifically limited. When the fixing part of the temperature sensing detector is connected to the reactor cover 6 at the temperature sensor probe port 2, the airtightness of the accommodation cavity 15 can be ensured. Preferably, the diameter of the temperature sensor probe port 2 can be 4 mm. When the directional microecological bioreactor is in use, the temperature sensing detector can continuously feedback the temperature of the culture medium in the accommodation cavity 15 of the reactor tank 7.

[0040] In one embodiment, the inlet of the sampling tube is located inside the reactor tank 7 and below the liquid level of the culture medium in the reactor tank 7; the outlet of the sampling tube is located outside the reactor tank 7 and is connected to a sample guiding tube 18, and a first on-off valve 14 is provided on the sample guiding tube 18; the fixing part of the sampling tube is detachably connected to the reactor cover 6 at the sampling tube opening 3. In the present disclosure, the connection method of the fixing part of the sampling tube detachably connected to the reactor cover 6 is not specifically limited. When the fixing part of the sampling tube is connected to the reactor cover 6 at the sampling tube opening 3, the airtightness of the accommodation cavity 15 can be ensured. Preferably, the diameter of the sampling tube opening 3 can be 4 mm. When the directional microecological bioreactor is in use, the culture in the reactor tank 7 can be sampled and detected through the sampling tube after a certain period of time, so as to know the fermentation degree of the sample in the reactor tank 7. The first on-off valve 14 provided on the sample guiding tube 18 is opened during sampling and closed when not sampling, thereby ensuring the airtightness of the accommodation cavity of the reactor tank 7.

[0041] In one embodiment, the outlet of the aeration tube is located inside the reactor tank 7 and at the bottom of the reactor tank 7; the inlet of the aeration tube is located outside the reactor tank 7 and is connected to an air guiding tube 12, and the inlet of the air guiding tube 12 is connected to an air pump 9; a bacteria filter 10, an air flow meter 11 and a second on-off valve 13 are arranged in sequence along the direction from the inlet to the outlet of the air guiding tube 12; the fixing part of the aeration tube is detachably connected to the reactor cover 6 at the aeration tube opening 4. In the present disclosure, the connection method of the fixing part of the aeration tube detachably connected to the reactor cover 6 is not specifically limited. When the fixing part of the aeration tube is connected to the reactor cover 6 at the aeration tube opening 4, the airtightness of the accommodation cavity 15 can be ensured. Preferably, the diameter of the aeration tube opening 4 can be 2 mm. When the directional microecological bioreactor is in use, oxygen required for fermentation can be introduced into the accommodation cavity of the reactor tank 7 through the air guiding tube 12; when oxygen is required for fermentation, the second on-off valve 13 is opened and the air pump 9 is started, so that air enters from the inlet of the air guiding tube 12 and bacteria and other substances are filtered out in the bacteria filter 10, and then enters the reactor tank 7; when oxygen does not need to be introduced for fermentation, the second on-off valve 13 is closed to ensure the airtightness of the accommodation cavity of the reactor tank 7. The material of the air guiding tube 12 can be silica gel, and the diameter of the air guiding tube 12 can be 2 mm.

[0042] In a preferred embodiment, the aeration pipe includes a gas inlet portion 16 and an aeration portion 17; wherein, the inlet of the inlet portion 16 (i.e., the inlet of the aeration pipe) is located outside the reactor tank body 7 and is connected to the outlet of the air guide pipe 12; the inlet portion 16 can be a pipeline with a height of 180 mm and a diameter of 2 mm; the inlet of the aeration portion 17 is connected to the outlet of the inlet portion 16, and gas can enter the aeration portion 17 through the inlet portion 16; the aeration portion 17 is a pipeline with an annular structure, and a plurality of aeration holes are provided on the aeration portion 17. As Figure 4 shown, The outer represents the outer diameter of the annular structure of the aeration portion 17, The inner represents the inner diameter of the annular structure of the aeration portion 17, The inner can be 54 mm, The outer can be 58 mm; H16 represents the height of the inlet portion 16, represents the diameter of the pipeline of the inlet portion 16, can be 2 mm. In the present disclosure, the aeration portion is located at the bottom of the reactor tank body, and oxygen can enter the reactor tank body from the aeration holes of the aeration portion. On the one hand, it provides sufficient oxygen for fermentation, and on the other hand, when gas enters the culture medium, it has gas disturbance and can play a stirring role. The aeration holes can be circular, with a pore diameter of 0.5 - 1.0 mm, preferably 0.5 mm; the number of the aeration holes can be 10 - 14. Preferably, the aeration holes are circular with a diameter of 0.5 mm, and one aeration hole is provided every 15 mm. Every 15 mm refers to the arc length between two adjacent aeration holes.

[0043] In one embodiment, a magnetic rotor 5 is further installed in the reactor tank body 7. Preferably, the number of the magnetic rotors 5 is 1. In the present disclosure, the magnetic rotor can be conventionally used by those skilled in the art, and will not be elaborated herein.

[0044] In one embodiment, a magnetic stirring constant temperature water bath 8 is further included for heating the reactor tank body 7. In the present disclosure, the magnetic stirring constant temperature water bath 8 can, on the one hand, heat the reactor tank body 7, which helps to maintain the temperature required for fermentation, for example, the temperature can be controlled at 20 - 100 °C; on the other hand, it can also control the rotation speed of the magnetic rotor 5 to ensure that the magnetic rotor gives an appropriate stirring rate to the culture medium. For example, the rotation speed of the magnetic rotor can be 0 - 800 rpm.

[0045] In one embodiment, the volume of the reactor tank body 7 is 300 - 500 mL. Preferably 500 mL.

[0046] A directional microecological bioreactor of the present disclosure, each component of the device is arranged as Figure 1For assembly, the directional microecological bioreactor includes a reactor tank body 7 and a reactor cover body 6;

[0047] The interior of the reactor tank body 7 is hollow and forms a receiving cavity 15 that is closed at one end and open at the other end; the reactor cover body 6 is detachably connected to the reactor tank body 7 for closing the open end of the receiving cavity 15;

[0048] A pressure valve port 1, a temperature sensor probe port 2, a sampling pipe port 3, and an air supply pipe port 4 are provided on the covering part of the reactor cover body 6; a pressure valve is provided at the pressure valve port 1, a temperature sensing detector is inserted into the temperature sensor probe port 2, a sampling pipe is inserted into the sampling pipe port 3, and an air supply pipe is inserted into the air supply pipe port 4.

[0049] The temperature measuring part of the temperature sensing detector is located inside the reactor tank body 7, and the fixing part of the temperature sensing detector is detachably connected to the reactor cover body 6 at the temperature sensor probe port 2.

[0050] The inlet of the sampling pipe is located inside the reactor tank body 7; the outlet of the sampling pipe is located outside the reactor tank body 7 and is connected to a sample guiding pipe 18, and a first straight-through valve 14 is provided on the sample guiding pipe 18; the fixing part of the sampling pipe is detachably connected to the reactor cover body 6 at the sampling pipe port 3.

[0051] The outlet of the air supply pipe is located inside the reactor tank body 7; the inlet of the air supply pipe is located outside the reactor tank body 7 and is connected to a gas guiding pipe 12, and the inlet of the gas guiding pipe 12 is connected to a gas guiding pump 9; a bacteria filter 10, an air flow meter 11, and a second straight-through valve 13 are arranged in sequence along the direction from the inlet to the outlet of the gas guiding pipe 12; the fixing part of the air supply pipe is detachably connected to the reactor cover body 6 at the air supply pipe port 4.

[0052] The air supply pipe includes a gas introduction part 16 and an aeration part 17. The aeration part 17 is a pipeline with an annular structure, and 14 circular aeration holes with a diameter of 0.5 mm are provided on the aeration part.

[0053] One magnetic rotor 5 is also installed inside the reactor tank body 7.

[0054] It also includes a magnetic stirring constant temperature water bath 8 for heating the reactor tank body 7.

[0055] The volume of the reactor tank body 7 is 350 mL. A culture medium is loaded into the reactor tank body 7, and the loading amount of the culture medium is 60% of the receiving cavity.

[0056] The following further details the present disclosure in conjunction with embodiments, but the scope of the present disclosure is not limited to the following embodiments.

[0057] Without special instructions, the raw materials used in the present disclosure are all commercially available products.

[0058] Example 1

[0059] As shown Figure 1 in the figure, the reactor tank body 7 (500 mL) is filled with a culture medium, and the filling amount of the culture medium is 60% (300 mL) of the accommodation cavity. The culture medium is a Bacillus subtilis liquid culture medium (formula: glucose 8.50 g / L, peptone 5.83 g / L, yeast extract 2.50 g / L, potassium dihydrogen phosphate 0.35 g / L, calcium carbonate 0.25 g / L). A magnetic rotor is placed in the reactor, the reactor cover body 6 is hermetically connected to the reactor tank body 7, and a pressure valve, a temperature sensing detector, a sampling tube and an aeration tube are assembled with the reactor cover body 6, and sterilized at 121 °C for 30 min by moist heat sterilization.

[0060] After sterilization, weighing and inoculating Bacillus subtilis is carried out according to the initial inoculation amount of 0.5×10 8 CFU / mL. Calculated according to the liquid loading amount of 300 mL, the required amount of Bacillus subtilis (bacterial amount: 10 11 CFU / g) is 0.15 g. After weighing, the bacterial powder of Bacillus subtilis is added into the directional microecological bioreactor, and after adding, tighten the cover. This process is completed in a clean room.

[0061] Place the reactor tank body 7 in the magnetic stirring constant temperature water bath 8, adjust the temperature to 37 °C, adjust the rotation speed to 300 rpm, connect the aeration tube 4 to the conduit, and open the bacterium filter 10, the air flow meter 11 and the second straight-through valve 13. Connect the power supply of the air pump, and adjust the air flow meter 11 to make the aeration gas volume 900 / min.

[0062] Sampling is carried out after culturing for 20 h: When sampling, just open the first straight-through valve 14 to directly sample, and close the straight-through valve 14 after sampling.

[0063] Dilute coating counting is carried out on the culture solution. The results are shown in Table 1. The initial inoculation amount is about 0.5×10 8 CFU / mL, which is basically consistent with the estimation. After 20 hours of incubation and aeration fermentation at 37 °C, the viable count of Bacillus subtilis is 45×10 8 CFU / mL.

[0064] Comparative Example 1

[0065] DME50 is a product of Beijing Lvdan Biotechnology Co., Ltd., and the national invention patent number is: ZL202310943775.1.

[0066] The same process parameters as in Example 1 are adopted, except that the fermentation is carried out in DME50. The results are shown in Table 1.

[0067] Table 1

[0068]

[0069] As can be seen from Table 1, the directed microecological bioreactor of the present disclosure can achieve an amplification of nearly 100 times that of Bacillus subtilis, and has a high consistency with the fermentation of DME50 in Comparative Example 1.

[0070] The preferred embodiments of the present disclosure have been described in detail above in conjunction with the accompanying drawings. However, the present disclosure is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present disclosure, various simple modifications can be made to the technical solutions of the present disclosure, and these simple modifications all fall within the protection scope of the present disclosure.

[0071] In addition, it should be noted that, in the above specific embodiments, the various specific technical features described can be combined in any suitable manner without contradiction. To avoid unnecessary repetition, the present disclosure will not separately describe various possible combination methods.

[0072] Furthermore, any combination can be made between the various different embodiments of the present disclosure, as long as it does not violate the idea of the present disclosure, and it should also be regarded as the content disclosed by the present disclosure.

Claims

1. A directional microecological bioreactor, characterized in that: The directional microecological bioreactor comprises a reactor tank (7) and a reactor cover (6); The reactor tank body (7) is hollow inside and forms a receiving cavity (15) with one end closed and the other end open; the reactor cover body (6) is detachably connected to the reactor tank body (7) and is used to close the open end of the receiving cavity (15); A pressure valve port (1), a temperature sensor probe port (2), a sampling tube port (3) and an aeration tube port (4) are arranged on the covering portion of the reactor cover body (6); a pressure valve is arranged at the pressure valve port (1), a temperature sensor probe is inserted into the temperature sensor probe port (2), a sampling tube is inserted into the sampling tube port (3), and an aeration tube is inserted into the aeration tube port (4).

2. The directional microecological bioreactor according to claim 1, characterized in that: The temperature measuring part of the temperature sensor detector is located in the reactor tank (7), and the fixing part of the temperature sensor detector is detachably connected to the reactor cover (6) at the temperature sensor probe port (2).

3. The directional microecological bioreactor according to claim 1, characterized in that: The inlet of the sampling tube is located in the reactor tank (7); The outlet of the sampling tube is located outside the reactor tank (7) and is connected to a sample guide tube (18), and a first straight valve (14) is provided on the sample guide tube (18); The fixing portion of the sampling tube is detachably connected to the reactor cover (6) at the sampling tube opening (3).

4. The directional microecological bioreactor according to claim 1, characterized in that: The outlet of the aeration pipe is located in the reactor tank (7); The inlet of the aeration pipe is located outside the reactor tank (7) and is connected to an air guide pipe (12), and the inlet of the air guide pipe (12) is connected to an air guide pump (9); a bacteria filter (10), an air flow meter (11) and a second through valve (13) are sequentially arranged along the direction from the inlet to the outlet of the air guide pipe (12); The fixing portion of the aeration pipe is detachably connected to the reactor cover (6) at the aeration pipe opening (4).

5. The directional microecological bioreactor according to claim 4, characterized in that: The aeration pipe comprises a gas introduction portion (16) and an aeration portion (17); the aeration portion (17) is a pipeline with an annular structure, and a plurality of aeration holes are arranged on the aeration portion (17).

6. The directional microecological bioreactor according to claim 5, characterized in that: The aeration holes are circular, with a hole diameter of 0.5-1.0 mm; the number of the aeration holes is 10-14.

7. The directional microecological bioreactor according to claim 1, characterized in that: The reactor tank (7) is also equipped with a magnetic rotor (5).

8. The directional microecological bioreactor according to claim 7, characterized in that: The number of the magnetic rotor (5) is one.

9. The directional microecological bioreactor according to claim 1, characterized in that: It also includes a magnetic stirring constant temperature water tank (8) for heating the reactor tank (7).

10. The directional microecological bioreactor according to claim 1, characterized in that: The volume of the reactor tank (7) is 300-500 mL.

Images