Stirring paddle and cell-free synthesis reaction device

Abstract

The utility model discloses a kind of stirring paddle and cell-free synthesis reaction device. Stirring paddle includes stirring shaft and at least one paddle of being arranged on the stirring shaft, the inside of the stirring shaft is provided with first gas passage, the inside of the paddle has a cavity, the first gas passage is communicated with the cavity inside the paddle, and, the paddle has gas permeation diffusion structure, gas in the cavity from the first gas passage can be slowly diffused to the outside of the paddle via the gas permeation diffusion structure. The stirring paddle provided in the utility model embodiment gives consideration to oxygen supply efficiency and bubble control, realizes the uniform dispersion of gas in liquid, avoids the formation of bubble, thereby solve the problem of bubble aggregation and overflow in conventional aeration mode.

Description

Technical Field

[0001] This utility model specifically relates to a stirring paddle and a cell-free synthesis reaction device, belonging to the field of bioreactor technology. Background Technology

[0002] Proteins, as potential therapeutic biopharmaceuticals, drug targets, and biocatalysts, hold significant scientific and practical importance. While existing technologies have enabled the successful expression and production of numerous proteins, many crucial proteins remain difficult to express effectively for various reasons, such as cytotoxic proteins and poorly soluble proteins. With advancements in science and technology, cell-free protein expression technology, due to its unique open system characteristics—allowing for control of reaction conditions, reactant components, and direct monitoring of reaction expression—has become a powerful protein expression method. Cell-free protein synthesis (CFPS) systems are in vitro gene expression systems that use exogenous DNA or mRNA as templates, artificially adding necessary raw materials and energy substances, and using cell extracts as conditions to synthesize proteins. This overcomes cellular limitations, enabling convenient and rapid expression of various proteins. Specifically, CFPS uses DNA as a template, and under the action of RNA polymerase and transcription factors, transcribes the corresponding mRNA; then, using mRNA as a template, it translates and synthesizes proteins using ribosomes, amino acid substrates, tRNA, and energy substances within the system. Cell-free protein expression does not require the maintenance of cell metabolism, making it more efficient. Due to its unique open system characteristics, it allows for control of reaction conditions, reactant components, and direct monitoring of reaction expression, and has the potential to express various toxic and difficult-to-express proteins.

[0003] In recent years, cell-free protein synthesis systems have demonstrated significant advantages in fields such as biomedicine, industrial enzyme production, and synthetic biology. However, with the expansion of application scenarios and the increasing demands of industrialization, these systems face numerous challenges in areas such as oxygen supply, reaction uniformity, and large-scale scaling. Oxygen is a core element for energy metabolism and redox balance in cell-free protein expression systems: on the one hand, as the terminal acceptor of the electron transport chain, it drives ATP regeneration through oxidative phosphorylation, maintaining the continuous progress of energy-consuming reactions such as ribosome translation and amino acid activation; on the other hand, oxygen directly participates in the regulation of oxidoreductase activity and disulfide bond formation, ensuring the correct folding and functional integrity of the target protein. However, the open nature of cell-free systems leads to a significantly higher oxygen consumption rate than traditional cell culture systems, and the lack of physical barriers such as cell membrane structures in the reaction solution directly affects the reaction process due to the incompatibility of oxygen and the efficiency of mass transfer.

[0004] Existing mature solutions mostly employ rigid stirring impellers (such as turbine or anchor impellers) combined with bottom gas distributors (such as microporous discs) to achieve mixing and aeration. However, this approach suffers from problems such as generating a large number of bubbles, requiring defoaming agents, uneven gas distribution, and low dissolved oxygen efficiency. For example, CN203200277U discloses a bioreactor with a hollow stirring shaft and several vents on its surface, featuring at least two stirrers to increase dissolved oxygen levels. However, the amount of oxygen provided by the vents on the stirring shaft is limited, making it difficult to meet the oxygen requirements of cell-free reactions. Furthermore, it still relies on bubble oxygen supply, which can lead to excessive bubble volume causing the reaction liquid to overflow and abnormally terminating the reaction. In particular, it suffers from the following drawbacks: a contradiction between oxygen supply efficiency and bubble control: traditional cell-free reaction systems require a high aeration rate strategy to achieve sufficient oxygen supply, but this operation mode leads to a dramatic increase in bubble generation, making it easy for the reaction liquid to overflow from the container and affecting the normal progress of the reaction. Limited gas-liquid mass transfer interface: traditional aeration methods generate large-diameter bubbles, resulting in a limited gas-liquid contact area, making the dissolved oxygen rate the rate-limiting step in the reaction process. Experiments show that while increasing ventilation rate can temporarily boost oxygen transfer, it shortens the effective gas-liquid contact time, thus reducing the volumetric mass transfer coefficient. Furthermore, increased reaction liquid viscosity promotes bubble coalescence, further reducing the mass transfer interface area. The use of defoamers presents a dilemma: current technologies largely rely on chemical defoamers to suppress foam, but high concentrations can inhibit target protein expression or interfere with cell metabolism, while low-concentration solutions are insufficient to effectively control foam formation.

[0005] In summary, while high aeration rates can alleviate mass transfer bottlenecks, they exacerbate foaming problems; and while the addition of defoamers can suppress foaming, it may introduce new protein expression inhibitors. This technical contradiction limits the application potential of cell-free reaction systems in large-scale production. Existing oxygen supply methods rely on a separate design of rigid impellers and independent aeration devices (such as microporous disks), which generates a large number of bubbles while improving dissolved oxygen efficiency. This can easily lead to overflow of the reaction solution, preventing the reaction from proceeding normally. At the same time, bubbles can interfere with the homogeneity of the reaction system, hindering sufficient contact between the substrate, template, and key components, thus reducing reaction efficiency. If fluorescence monitoring is used, bubbles can also cause signal distortion, affecting data accuracy. Furthermore, bubbles may clog microfluidic devices or channels, triggering false alarms, interfering with automated operation processes, and in high-concentration systems, they may cause cascade protein inactivation, ultimately leading to problems such as decreased product yield, reduced activity, and poor experimental reproducibility. Utility Model Content

[0006] The main purpose of this invention is to provide a stirring paddle and a cell-free synthesis reaction device, thereby overcoming the shortcomings of the prior art.

[0007] To achieve the aforementioned objectives, the technical solution adopted by this utility model includes:

[0008] A first aspect of this utility model provides a stirring paddle, including a stirring shaft and at least one blade disposed on the stirring shaft. The stirring shaft has a first gas channel inside, and the blade has a cavity inside. The first gas channel communicates with the cavity inside the blade, and the blade has a gas permeation and diffusion structure, through which gas entering the cavity from the first gas channel can slowly diffuse to the outside of the blade.

[0009] A second aspect of this utility model provides a cell-free synthetic reaction apparatus, comprising: a stirring paddle, a reaction container, a driving mechanism, and a gas source. The stirring paddle is disposed within the reaction container, the driving mechanism is throttle-connected to the stirring paddle and is used to drive the stirring paddle to rotate, and the gas source is connected to a first gas channel within the stirring paddle and is used to provide gas.

[0010] Compared with the prior art, the advantages of this utility model include:

[0011] The stirring paddle provided in this embodiment of the invention balances oxygen supply efficiency and bubble control, achieving uniform dispersion of gas in the liquid and avoiding bubble formation, thereby solving the problems of bubble aggregation and overflow in traditional aeration methods.

[0012] The stirring paddle provided in this embodiment improves the gas-liquid mass transfer efficiency. By utilizing the gas permeation and diffusion structure material on the paddle blades, the gas can be uniformly permeated into the reaction liquid, significantly increasing the gas-liquid contact area and dissolved oxygen rate. This overcomes the problem of low gas-liquid mass transfer efficiency in traditional aeration methods and meets the high oxygen consumption requirements of CFPS.

[0013] The stirring paddle provided in this embodiment of the invention avoids the generation of foam by improving the gas transmission structure / method, thereby eliminating the dependence on chemical defoamers and further reducing production costs. Attached Figure Description

[0014] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments recorded in this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0015] Figure 1 This is a schematic diagram of a cell-free synthetic reaction device based on a stirring paddle provided in a typical embodiment of this utility model;

[0016] Figure 2 This is a schematic diagram of the internal structure of the blade in a typical embodiment of this utility model;

[0017] Figure 3 This is a schematic diagram of the internal structure of a stirring paddle provided in a typical embodiment of this utility model;

[0018] Figure 4 , Figure 5 This is a schematic diagram of the structure of a stirring paddle with a single spiral blade provided in a typical embodiment of this utility model;

[0019] Figure 6 This is a top view of a single helical blade and its internal air passage provided in a typical embodiment of this utility model;

[0020] Figure 7 This is a schematic diagram of the internal structure of the spiral blade provided in a typical embodiment of this utility model;

[0021] Figure 8 This is a top view of a single helical blade provided in a typical embodiment of this utility model. Detailed Implementation

[0022] In view of the shortcomings of the prior art, the inventor of this case, through long-term research and extensive practice, has come up with the technical solution of this utility model. The following will further explain the technical solution, its implementation process, and its principles.

[0023] A first aspect of this utility model provides a stirring paddle, including a stirring shaft and at least one blade disposed on the stirring shaft. The stirring shaft has a first gas channel inside, and the blade has a cavity inside. The first gas channel communicates with the cavity inside the blade, and the blade has a gas permeation and diffusion structure, through which gas entering the cavity from the first gas channel can slowly diffuse to the outside of the blade.

[0024] Furthermore, the internal cavity of the blade is provided with one or more second gas channels, each with multiple gas outlets. The second gas channel is connected to the cavity via the gas outlets, and the second gas channel is directly connected to the first gas channel. The first gas channel is connected to the cavity only via the second gas channel.

[0025] Furthermore, the multiple gas outlets on each of the second gas channels are evenly distributed circumferentially along the length of the second gas channel.

[0026] Furthermore, a second gas channel is provided in the cavity inside the blade, and the second gas channel has a wave-shaped or spiral structure.

[0027] Alternatively, the internal cavity of the blade may be provided with multiple second gas channels, which are radially distributed with the connection point with the first gas channel as the center. Alternatively, one selected second gas channel may be directly connected to the first gas channel, and the remaining multiple second gas channels may be spaced apart along the length of the selected second gas channel and connected to the selected second gas channel respectively. The second gas channel may be a straight cylindrical, wavy, or spiral structure.

[0028] In a more specific embodiment, the stirring paddle includes a plurality of blades, and the cavities inside the plurality of blades are independently connected to the first gas channel, or the cavities inside the plurality of blades are connected sequentially, and the first gas channel is directly connected to the cavity inside at least one of the blades.

[0029] Furthermore, the second gas channels inside the multiple blades are independently connected to the first gas channel, or the second gas channels inside the multiple blades are connected sequentially.

[0030] Furthermore, the multiple blades are combined into multiple blade groups, each blade group includes at least two blades, the blades in the same blade group are located at the same axial position of the stirring shaft, and the multiple blade groups are spaced apart along the axial direction of the stirring shaft.

[0031] In a more specific implementation, a third gas channel is further provided inside the stirring shaft. The third gas channel is connected to the cavity inside the impeller. The first gas channel serves as an inlet channel, and the third gas channel serves as an outlet channel. The first gas channel and the third gas channel are arranged in parallel. The inlet at the beginning of the first gas channel and the outlet at the end of the third gas channel are located at the same end. Gas supplied by an external gas source enters the first gas channel through the inlet, enters the cavity inside the impeller through the second gas channel, diffuses some of the gas through the gas permeation and diffusion structure on the impeller to the outside of the impeller, and the remaining gas is discharged through the outlet of the third gas channel.

[0032] In a more specific embodiment, the agitator includes: a first vent pipe, a second vent pipe, and a third vent pipe. The first vent pipe and the third vent pipe are disposed inside the agitator shaft. The first gas channel is disposed inside the first vent pipe, and the third gas channel is disposed inside the third vent pipe. The second vent pipe is disposed in the cavity inside the blade, and the second gas channel is disposed inside the second vent pipe.

[0033] Furthermore, the blade includes a support frame and a gas-permeable membrane covering the support frame, the support frame and the gas-permeable membrane together forming the cavity.

[0034] Furthermore, the blades are helical blades.

[0035] Furthermore, the spiral blade has a spiral radius of 0.5cm-1.4cm, a pitch of 1.2cm-5.5cm, and a spiral angle of 25°-45°.

[0036] Furthermore, the central axis of the spiral blade is parallel or perpendicular to the central axis of the stirring shaft.

[0037] A second aspect of this utility model provides a cell-free synthetic reaction apparatus, comprising: a stirring paddle, a reaction container, a driving mechanism, and a gas source. The stirring paddle is disposed within the reaction container, the driving mechanism is throttle-connected to the stirring paddle and is used to drive the stirring paddle to rotate, and the gas source is connected to a first gas channel within the stirring paddle and is used to provide gas.

[0038] The following will further explain the technical solution, its implementation process and principle in conjunction with the accompanying drawings and specific implementation examples. Unless otherwise specified, the air source, motor and other components involved in the embodiments of this utility model are known in the art and are not specifically limited here.

[0039] In a typical implementation, a cell-free synthesis reaction device based on a stirrer includes a stirrer, a reaction container 10, a drive mechanism, and a gas source. The stirrer is disposed inside the reaction container 10. The drive mechanism is connected to the stirrer and is used to drive the stirrer to rotate. The gas source is connected to a first gas channel 410 inside the stirrer and is used to provide gas.

[0040] Specifically, the reaction vessel 10 is used to contain the reaction liquid. The drive mechanism and the gas source are located outside the reaction vessel 10. For example, the volume of the reaction vessel 10 can be 15 mL. The specific structure of the reaction vessel 10 is not an improvement of this utility model, and its structure can be known in the art and is not specifically limited here. Specifically, the drive mechanism can include a motor, etc. Of course, the drive mechanism can also include other auxiliary accessories that enable the motor to drive the stirring paddle, etc. These are all conventional structures known in the art and are not specifically limited here. For example, the rotation speed of the stirring paddle during operation can be 50 rpm to 200 rpm. Specifically, the gas source can be a gas storage container and an air pump, etc. The structure and model of the gas storage container and the air pump are not limited.

[0041] Specifically, as the main improvement of this utility model, the structure and advantages of the stirring paddle will be described in detail below.

[0042] Please refer to the following: Figure 1 and Figure 2 The stirring paddle includes a stirring shaft 20 and one or more blades 30 mounted on the stirring shaft 20. The stirring shaft 20 is rotatably coupled to the reaction vessel 10, specifically through a coupling / mechanical seal structure, etc., to achieve both sealing and rotatable coupling. The stirring shaft 20 is connected to a drive mechanism, and all the blades 30 are located inside the reaction vessel 10. Specifically, each blade 30 has a cavity inside and a gas permeation and diffusion structure. The stirring shaft 20 has parallel first gas channels 410 and third gas channels 430 inside. The first gas channel 410 serves as an inlet channel directly connected to a gas source, and the third gas channel 430 serves as an outlet channel. The first gas channels 410 and third gas channels 430 are respectively connected to the cavities inside the blades 30. Gas supplied by the gas source enters the cavity through the first gas channel 410. Part of the gas entering the cavity slowly diffuses to the reaction liquid outside the blades 30 via the gas permeation and diffusion structure, while the other part returns to the gas storage container of the gas source via the third gas channel 430. Through this design, the impeller 30 with a gas permeation and diffusion structure achieves uniform dispersion of gas in the liquid, avoiding the formation of bubbles, thus solving the problems of bubble aggregation and overflow in traditional aeration methods. In addition, the gas permeation and diffusion structure enables gas to permeate evenly into the reaction liquid, significantly improving the gas-liquid contact area and dissolved oxygen rate, thereby overcoming the problem of low gas-liquid mass transfer efficiency in traditional aeration methods and meeting the high oxygen consumption requirements of CFPS.

[0043] Please refer to the following for details. Figure 1 and Figure 3 The stirring shaft 20 can be hollow. The first gas channel 410 and the third gas channel 430 can be integrally formed with the stirring shaft 20. Of course, the stirring shaft 20 can also be additionally provided with a first vent pipe and a third vent pipe. The cavities inside the first vent pipe and the third vent pipe serve as the first gas channel 410 and the third gas channel 430, respectively. The end ports of the first gas channel 410 and the third gas channel 430 near the top of the stirring shaft 20 (the connection end between the stirring shaft 20 and the drive mechanism is the top) serve as the air inlet 4001 and the air outlet 4002, respectively. That is, the air inlet 4001 and the air outlet 4002 of the gas passage 40 formed by the first gas channel 410, the third gas channel 430 and the cavities inside the multiple blades 30 are located at the same end.

[0044] Specifically, the cavities inside the multiple blades 30 are located between the first gas channel 410 and the third gas channel 430 along the gas flow direction within the gas passage 40. In a first embodiment, the cavities inside the multiple blades 30 can be connected in series, that is, the cavities inside the multiple blades 30 are connected end-to-end sequentially, with one cavity at the beginning directly connected to the first gas channel 410, and one cavity at the end directly connected to the third gas channel 430 (e.g., ...). Figure 1 , Figure 3 (As shown). In a second embodiment, the cavities inside the multiple blades 30 are connected in parallel, that is, the cavities inside the multiple blades 30 are simultaneously connected to the first gas channel 410 and the third gas channel 430. In a third embodiment, the cavities inside the multiple blades 30 are connected by a combination of series and parallel connections. This embodiment includes many specific connection structures, which will not be described one by one here.

[0045] For details, please refer to the following document again. Figure 2 , Figure 7 To improve the uniformity of gas permeation / diffusion from the interior of the blade 30, one or more second gas channels 3002 are provided in the cavity inside the blade 30. Each second gas channel 3002 has multiple gas outlets. The second gas channel 3002 communicates with the cavity via these gas outlets. The second gas channel 3002 is directly connected to the first gas channel 410, which is connected to the cavity only via the second gas channel 3002. Similarly, a third gas channel 430 is also connected to the second gas channel 3002. That is, gas exits sequentially through the first gas channel 410, the second gas channel 3002, and the third gas channel 430. When passing through the second gas channel 3002, the gas enters the cavity inside the blade 30 through the gas outlets on the second gas channel 3002. Specifically, to ensure the uniformity of gas permeation / diffusion from the interior of the blade 30, the multiple gas outlets on each second gas channel 3002 are evenly distributed circumferentially along the length of the second gas channel 3002. It should be noted that the connection relationship of the second gas channels 3002 inside the multiple blades 30 is the same as the connection relationship between the cavities inside the multiple blades 30, that is, the second gas channels 3002 inside each blade 30 can be connected in series, in parallel or in a series-parallel combination.

[0046] It should be noted that the second gas passage 3002 can be integrally formed with the blade 30, or the blade 30 can have a second vent pipe inside, with the cavity of the second vent pipe serving as the second gas passage 3002.

[0047] Specifically, to ensure the uniformity of the gas permeating / diffused from the inside of the blade 30, a second gas channel 3002 is provided in the cavity inside each blade 30. The second gas channel 3002 has a wavy or spiral structure. Alternatively, multiple second gas channels 3002 are provided in the cavity inside the blade 30. The multiple second gas channels 3002 are radially distributed with the connection point with the first gas channel 410 as the center. Alternatively, one selected second gas channel 3002 is directly connected to the first gas channel 410, and the remaining multiple second gas channels 3002 are spaced apart along the length direction of the selected second gas channel 3002 and are respectively connected to the selected second gas channel 3002. The second gas channel 3002 has a straight cylindrical, wavy, or spiral structure.

[0048] Specifically, multiple blades 30 are combined into multiple blade 30 groups, each blade 30 group includes at least two blades 30, and the blades 30 included in the same blade 30 group are located at the same axial position of the stirring shaft 20. Multiple blade 30 groups are spaced apart along the axial direction of the stirring shaft 20. Specifically, the gas permeation diffusion structure may be distributed only in a local area on the surface of the blade 30, preferably covering the entire surface of the blade 30. Specifically, the blade 30 includes a shell 3001 and a cavity formed by the shell. The shell 3001 may include a supporting frame and a gas permeation membrane covering the supporting frame. The gas permeation membrane is preferably made of a breathable polymer material, and the breathable polymer material is selected from at least one of polydimethylsiloxane (PDMS), polyurethane (PU), Teflon AF-2400, porous silica gel, and modified polyurethane.

[0049] Specifically, such as Figures 4-8 As shown, the stirring paddle includes a helical blade 30. The helical blade has a helical radius of 0.5cm-1.4cm, a pitch of 1.2cm-5.5cm, and a helical angle of 25°-45°. The central axis of the helical blade is parallel to the central axis of the stirring shaft 20. The thickness of the blade 30 is 0.1-0.6mm, and the diameter of the blade can be 0.8cm. Figure 4 and Figure 5 The main difference between the impellers shown is the orientation and structure of the second gas channel 3002 inside the impeller blades.

[0050] The stirring paddle provided in this embodiment solves the problems of low oxygen supply efficiency, difficulty in bubble control, and difficulty in using defoamers in traditional cell-free reaction systems. It realizes a cell-free synthesis reaction device with high efficiency, bubble-free aeration, and integrated stirring structure, providing a new solution for the large-scale application of cell-free protein synthesis technology.

[0051] In traditional cell-free reaction systems, a high aeration rate strategy is typically employed to meet high oxygen demand. However, this operating mode easily leads to the generation of a large number of bubbles, causing problems such as reaction liquid overflow and decreased mass transfer efficiency. The stirring paddle provided in this embodiment of the invention balances oxygen supply efficiency and bubble control, achieving uniform gas dispersion in the liquid and avoiding bubble formation, thereby solving the problems of bubble aggregation and overflow in traditional aeration methods.

[0052] Traditional aeration methods generate large-diameter bubbles with limited gas-liquid contact area, making the dissolved oxygen rate the rate-limiting step in the reaction process. The stirring impeller provided in this embodiment improves gas-liquid mass transfer efficiency. By utilizing the gas permeation and diffusion structure material on the impeller blades, gas can uniformly permeate into the reaction liquid, significantly increasing the gas-liquid contact area and dissolved oxygen rate. This overcomes the problem of low gas-liquid mass transfer efficiency in traditional aeration methods and meets the high oxygen demand of CFPS.

[0053] Existing technologies often rely on chemical defoamers to suppress foam, but high concentrations of defoamers may inhibit target protein expression, while low concentrations are difficult to effectively control foam formation. The stirring paddle provided in this embodiment of the invention, by improving the gas transmission structure / method, avoids foam generation, thereby eliminating dependence on chemical defoamers and further reducing production costs.

[0054] It should be understood that the above embodiments are merely illustrative of the technical concept and features of this utility model, and are intended to enable those skilled in the art to understand the content of this utility model and implement it accordingly. They should not be construed as limiting the scope of protection of this utility model. All equivalent changes or modifications made in accordance with the spirit and essence of this utility model should be included within the scope of protection of this utility model.

Claims

1. A stirring impeller, comprising a stirring shaft and at least one impeller blade disposed on the stirring shaft, characterized in that: The stirring shaft has a first gas channel inside, and the blade has a cavity inside. The first gas channel communicates with the cavity inside the blade. Furthermore, the blade has a gas permeation and diffusion structure, so that the gas entering the cavity from the first gas channel can slowly diffuse to the outside of the blade through the gas permeation and diffusion structure.

2. The stirring paddle according to claim 1, characterized in that: The internal cavity of the blade is further provided with one or more second gas channels, each with multiple gas outlets. The second gas channel is connected to the cavity via the gas outlets. The second gas channel is directly connected to the first gas channel, while the first gas channel is connected to the cavity only via the second gas channel.

3. The stirring paddle according to claim 2, characterized in that: The multiple gas outlets on each of the second gas channels are evenly distributed circumferentially along the length of the second gas channel.

4. The stirring paddle according to claim 2 or 3, characterized in that: A second gas channel is provided in the cavity inside the blade, and the second gas channel has a wave-shaped or spiral structure. Alternatively, the internal cavity of the blade may be provided with multiple second gas channels, which are radially distributed with the connection point with the first gas channel as the center. Alternatively, one selected second gas channel may be directly connected to the first gas channel, and the remaining multiple second gas channels may be spaced apart along the length of the selected second gas channel and connected to the selected second gas channel respectively. The second gas channel may be a straight cylindrical, wavy, or spiral structure.

5. The stirring paddle according to claim 2, characterized in that: The stirring paddle includes a plurality of blades, and the cavities inside the plurality of blades are independently connected to the first gas channel, or the cavities inside the plurality of blades are connected sequentially, and the first gas channel is directly connected to the cavity inside at least one of the blades. And / or, the second gas channels inside the plurality of blades are independently connected to the first gas channel, or the second gas channels inside the plurality of blades are sequentially connected; And / or, the plurality of blades are combined into a plurality of blade groups, each blade group including at least two blades, the blades included in the same blade group being located at the same axial position of the stirring shaft, and the plurality of blade groups being spaced apart along the axial direction of the stirring shaft.

6. The stirring paddle according to claim 2 or 5, characterized in that: The stirring shaft is also provided with a third gas channel, which is connected to the cavity inside the impeller. The first gas channel serves as an inlet channel, and the third gas channel serves as an outlet channel. The first gas channel and the third gas channel are arranged in parallel. The inlet at the beginning of the first gas channel and the outlet at the end of the third gas channel are located at the same end. Gas supplied by an external gas source enters the first gas channel through the inlet, enters the cavity inside the impeller through the second gas channel, and part of the gas diffuses to the outside of the impeller through the gas permeation and diffusion structure on the impeller. The remaining gas is discharged through the outlet of the third gas channel.

7. The stirring paddle according to claim 6, characterized in that, include: A first vent pipe, a second vent pipe, and a third vent pipe are provided. The first vent pipe and the third vent pipe are disposed inside the stirring shaft. The first gas channel is disposed inside the first vent pipe, and the third gas channel is disposed inside the third vent pipe. The second vent pipe is disposed in the cavity inside the impeller, and the second gas channel is disposed inside the second vent pipe.

8. The stirring paddle according to claim 1, 2, or 5, characterized in that: The blade includes a support frame and a gas permeable membrane covering the support frame, and the support frame and the gas permeable membrane together enclose the cavity.

9. The stirring paddle according to claim 1, 2, or 5, characterized in that: The blades are helical blades; And / or, the helical blade has a helical radius of 0.5cm-1.4cm, a pitch of 1.2cm-5.5cm, and a helical angle of 25°-45°. And / or, the central axis of the spiral blade is parallel to or perpendicular to the central axis of the stirring shaft.

10. A cell-free synthetic reaction apparatus, characterized in that, include: The stirring paddle according to any one of claims 1-9, and a reaction vessel, a drive mechanism, and a gas source, wherein the stirring paddle is disposed inside the reaction vessel, the drive mechanism is throttle-connected to the stirring paddle and is used to drive the stirring paddle to rotate, and the gas source is connected to a first gas channel inside the stirring paddle and is used to provide gas.

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