WO2023098697A2 - 体外生物合成的反应装置以及体外生物合成方法 - Google Patents

体外生物合成的反应装置以及体外生物合成方法 Download PDF

Info

Publication number
WO2023098697A2
WO2023098697A2 PCT/CN2022/135274 CN2022135274W WO2023098697A2 WO 2023098697 A2 WO2023098697 A2 WO 2023098697A2 CN 2022135274 W CN2022135274 W CN 2022135274W WO 2023098697 A2 WO2023098697 A2 WO 2023098697A2
Authority
WO
WIPO (PCT)
Prior art keywords
reaction
chamber body
reaction chamber
partition
exhaust
Prior art date
Application number
PCT/CN2022/135274
Other languages
English (en)
French (fr)
Other versions
WO2023098697A3 (zh
Inventor
郭敏
陈文芳
于雪
Original Assignee
康码（上海）生物科技有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from CN202210095041.8A external-priority patent/CN114307940A/zh
Application filed by 康码（上海）生物科技有限公司 filed Critical 康码（上海）生物科技有限公司
Publication of WO2023098697A2 publication Critical patent/WO2023098697A2/zh
Publication of WO2023098697A3 publication Critical patent/WO2023098697A3/zh

Links

Images

Definitions

  • the invention belongs to the field of biotechnology, and in particular relates to a reaction device for in vitro biosynthesis and an in vitro biosynthesis method.
  • in vitro biosynthesis is compared to the synthesis in cells, such as in vitro cell-free protein synthesis (in vitro protein synthesis, in vitro cell-free protein synthesis): mainly rely on cell-free expression system, which is based on exogenous target DNA as protein Synthetic templates are artificially controlled to add substances such as substrates required for protein synthesis and cofactors for transcription and translation-related proteins, that is, the in vitro synthesis of the target protein is achieved through the reaction solution.
  • in vitro cell-free protein synthesis in vitro protein synthesis, in vitro cell-free protein synthesis
  • the purpose of the present invention is to solve the problem that the current in vitro biosynthesis cannot be scaled up. Therefore, the present invention provides the following technical solutions.
  • the invention provides a reaction device for in vitro biosynthesis, which is characterized in that it comprises: a reaction chamber body, the reaction chamber body is used to accommodate a synthetic reaction liquid for in vitro biosynthesis reaction, the reaction chamber body is provided with an opening, the opening The part has a feed port for allowing the synthesis reaction liquid to enter the reaction chamber body and a discharge port for discharging the reacted product; the partition part is used to incompletely separate the internal space of the reaction chamber body.
  • the reaction device provided by the present invention also has such a feature: wherein, the reaction chamber body is a tank body, preferably a horizontal tank body, and the tank body is spherical or ellipsoidal.
  • the reaction device provided by the present invention also has such a feature: wherein, the partition part includes at least one partition, preferably, the partition is in the shape of a sheet.
  • the reaction device provided by the present invention also has the feature that a plurality of partitions are evenly spaced.
  • the reaction device provided by the present invention also has such a feature: wherein, the length of the separator extends in a curved line or a broken line.
  • the reaction device provided by the present invention also has such a feature: wherein, there is an included angle between the line formed between the two ends of the length of the partition and the central axis of the inner space of the reaction chamber body whose length extends in the same direction. a.
  • the included angle a is less than or equal to 60°, preferably less than or equal to 45°, more preferably less than or equal to 20°.
  • the reaction device provided by the present invention also has such a feature: wherein at least one partition is rotatably and/or non-rotatably fixedly arranged on the inner wall of the reaction chamber body.
  • the reaction device provided by the present invention also has such a feature that when the partition is arranged on the inner wall, one side of the sheet-shaped partition faces the inner wall of the reaction chamber body.
  • the reaction device provided by the present invention also has such a feature: when the partition is arranged on the inner wall, one side of the sheet-shaped partition faces the inner wall of the reaction chamber body, and the other side of the partition away from the inner wall is set There are sunken areas.
  • the reaction device provided by the present invention also has such a feature that when one side of the sheet-shaped partition faces the inner wall of the reaction chamber body, the angle between the sheet-shaped surface of the partition and the opposite inner wall is not equal to 90°.
  • the reaction device provided by the present invention also has the feature that, in use, the reaction is carried out by rotating the reaction chamber body, preferably, the horizontal center axis of the reaction chamber body deviates from the horizontal state.
  • the reaction device provided by the present invention also has such a feature: wherein, at least one flow hole is arranged on the separator. Preferably, the flow holes in at least one partition are evenly distributed.
  • the reaction device provided by the present invention also has such a feature: when the partition is arranged on the inner wall, there is at least one discontinuity between the partition and the inner wall.
  • the discontinuities corresponding to at least one partition are equally spaced.
  • the reaction device provided by the present invention also has such a feature: wherein, the opening part also has a ventilation unit, and the ventilation unit has an air inlet and/or an exhaust port, and the air inlet is used to allow external gas to enter the reaction chamber body
  • the internal space, the exhaust port is used to discharge the gas in the internal space, preferably, one or more of the feed port, the discharge port, the air inlet and the exhaust port are the same opening.
  • the reaction device provided by the present invention also has such a feature, and further includes: an air intake unit and/or an exhaust unit, wherein the air inlet allows the outside gas to enter the internal space through the air intake unit, and the exhaust port passes through the air intake unit.
  • the unit lets the air out of the interior space exhaust the unit.
  • the reaction device provided by the present invention also has such features: wherein, the air intake unit includes at least one air intake pipe, one end of the air intake pipe faces the air inlet to deliver outside gas to the inner space, or one end of the air intake pipe passes from the inlet to the inner space.
  • the air port stretches into the inner space, at this time: one preferably, let at least one inlet pipe extend along the horizontal direction of the reaction chamber body; another preferably, at least one inlet pipe enters the part of the inner space on the pipe wall At least one air outlet hole, preferably, the at least one air outlet hole is evenly distributed, and/or one end of the air inlet pipe entering the inner space is sealed.
  • the exhaust unit includes an exhaust pipe, preferably, a part is arranged in the exhaust pipe between the two ends of the air inlet pipe, and one end of the air inlet pipe is separated from the exhaust pipe. One end protrudes into the inner space, and the other end of the air intake pipe passes through the other end of the exhaust pipe to communicate with the gas source.
  • the reaction device provided by the present invention also has such characteristics: it also includes: a cover body assembly, and the cover body assembly has a cover body adapted to cover the feed port with the feed port.
  • the lid assembly further includes a sealing ring, a lid lock and a locking handle.
  • the reaction device provided by the present invention also has such a feature: wherein, when the cover assembly is included, the exhaust unit also includes an exhaust hole, the exhaust hole is arranged on the cover, and the exhaust pipe passes through the exhaust hole and the inner space connected.
  • the reaction device provided by the present invention also has such a feature, and further includes: a rotating part, which is used to rotate the reaction chamber body, preferably, the rotating part rotates the reaction chamber body by being rotatably connected with the reaction chamber body, and/or rotates
  • the part is also provided with a runner, the reaction chamber body is arranged on the runner, and the runner rotates with the rotation of the reaction chamber body.
  • the reaction device provided by the present invention also has such a feature: wherein, the rotating part includes a driving unit, and the reaction chamber body is driven to rotate by the driving unit.
  • the reaction device provided by the present invention also has such a feature, and further includes: a temperature regulating device for keeping the reaction at a predetermined temperature; preferably, the temperature regulating device is used for controlling the gas entering the reaction chamber body. Tempering to maintain predetermined temperature conditions.
  • the reaction device provided by the present invention also has such a feature, and further includes: a housing, the housing is adapted to the reaction chamber body and is used to accommodate the reaction chamber body,
  • the housing has any one or a combination of the following features:
  • the housing is provided with an opening and closing door, preferably, the opening and closing door is provided with a ventilation hole;
  • the outer side of the housing is provided with a display screen, preferably, the display screen is located on the outer surface of the top of the housing;
  • the outer side of the housing is provided with a storage platform for placing articles.
  • the storage surface of the storage platform is provided with anti-skid lines, and preferably, the outer surface of the upper top of the housing is a storage platform;
  • the casing is provided with any one or more of the following: an air inlet passage for allowing external air to enter the inner space of the reaction chamber body, an exhaust passage for allowing the gas in the inner space of the reaction chamber body to discharge,
  • the reaction liquid for synthesis enters the inner space of the reaction chamber body to carry out the feed channel for the in vitro biosynthesis reaction, and the discharge channel for discharging the reacted product.
  • the present invention also provides an in vitro biosynthesis method, which is characterized in that: the aforesaid reaction device is used to accommodate the synthesis reaction solution for in vitro biosynthesis.
  • the in vitro biosynthesis method provided by the present invention also has the feature that the volume of the synthetic reaction solution used for the reaction accounts for less than or equal to 30% of the volume of the reaction chamber body.
  • the in vitro biosynthesis method provided by the present invention also has the feature that during the reaction process, the external gas is introduced into the internal space, and/or the gas in the internal space is exhausted.
  • the in vitro biosynthesis method provided by the present invention also has the feature that, during the reaction, the reaction chamber body of the reaction device is rotated, preferably, the rotation speed is less than or equal to 120r/min.
  • the invention provides a bioreactor, which is used for in vitro biosynthesis reaction, and is characterized in that it comprises: a reaction chamber body, which is used for accommodating a synthetic reaction solution for in vitro biosynthesis reaction, and an opening is arranged on the reaction chamber body , the opening has a feed inlet, a feed outlet and a ventilation unit.
  • the feed inlet is used to allow the reaction liquid for synthesis to enter the reaction chamber body
  • the discharge outlet is used to allow the product obtained after the reaction to be discharged from the reaction chamber body.
  • the unit has an air inlet and/or an exhaust port, the air inlet is used to allow external gas to enter the internal space of the reaction chamber body, and the exhaust port is used to discharge the gas in the internal space, preferably, the reaction chamber body is a tank .
  • the bioreactor provided by the present invention also has the feature that at least one first protrusion is arranged on the inner wall of the reaction chamber body.
  • the bioreactor provided by the present invention also has such a feature: wherein, one or more of the feed port, the feed port, the gas inlet and the exhaust port are the same opening.
  • the bioreactor provided by the present invention also has such a feature, and further includes: a rotating installation part, which is used for rotating connection with the rotating part of the rotating reaction chamber body, preferably, the rotating installation part is located on the opposite side of the opening , when the connection is rotated, the opening is inclined upward.
  • the present invention also provides a partition for biological reaction, which is characterized in that it is used for incompletely partitioning the internal space of the reaction chamber body of the aforementioned bioreactor.
  • the separator provided by the present invention also has the feature that the length of the separator extends in a curved line or a broken line.
  • the spacer provided by the present invention also has such a feature: wherein, the spacer is sheet-shaped, has two opposite sheet-shaped surfaces and side edges on both sides of the two opposite sheet-shaped surfaces.
  • the separator provided by the present invention also has the feature that a recessed area is provided on one side of the separator.
  • the separator provided by the present invention also has the feature that at least one flow hole is arranged on the separator.
  • the separator provided by the present invention also has the feature that at least one second protrusion is arranged on the outer surface of the separator.
  • the separator provided by the present invention also has the feature that the separator is integrally formed with the bioreactor.
  • the present invention also provides a shell, which is characterized in that: the shell is adapted to the reaction chamber body and is used for accommodating the reaction chamber body.
  • the housing provided by the present invention has any one or a combination of the following features:
  • the housing is provided with an opening and closing door, preferably, the opening and closing door is provided with a ventilation hole;
  • the outside of the housing is provided with a display screen, preferably, the display screen is located on the outer surface of the top of the housing;
  • the outer side of the housing is provided with a storage platform for placing articles.
  • the storage surface of the storage platform is provided with anti-skid lines, and preferably, the outer surface of the upper top of the housing is a storage platform;
  • the casing is provided with any one or more of the following: an air inlet channel for allowing external gas to enter the inner space of the reaction chamber body, an air outlet channel for allowing the gas in the inner space of the reaction chamber body to be discharged, and a gas outlet channel for the synthesis
  • the present invention also provides a reaction device, which is characterized in that it includes: a bioreactor and at least one partition, wherein at least one of the partitions incompletely separates the internal space of the reaction chamber body of the bioreactor.
  • the reactor and the partition are integrally formed, wherein the bioreactor is the aforementioned bioreactor, and the partition is the aforementioned partition.
  • the reaction device provided by the present invention also has the following features, and further includes: the aforementioned casing.
  • in vitro biosynthesis in the present invention mainly refers to in vitro biosynthesis such as in vitro protein synthesis reaction and in vitro mRNA synthesis reaction.
  • reaction liquid in the "flowing state" can flow from one space area to another The space area flows, and the reaction solution flowing through the partition will be blocked by the partition to increase the mixing intensity of the reaction solution during the in vitro biosynthesis reaction, thereby improving the reaction efficiency, and the overall structure is simple, and can be made of stainless steel, which is convenient for expansion chemical manufacturing and production;
  • the partition is sheet-shaped, which increases the contact area of the reaction solution, and also improves the degree of dilution of the reaction solution;
  • the partition is extended by a curve or a broken line.
  • the contact area of the reaction solution is also increased, which is beneficial to the dilution of the reaction solution;
  • the air inlet is used to allow external gas to enter the inner space of the reaction chamber body, so that oxygen-containing gas, such as air, can be introduced to facilitate in vitro biosynthesis reactions;
  • the exhaust port is used to discharge the gas in the internal space, so that the unfavorable gas generated in the reaction process, such as ethanol, etc., can be discharged in time to avoid affecting the synthesis efficiency and quality;
  • Fig. 1 is the overall structure schematic diagram of the first reaction device involved in the present invention
  • Fig. 2 is a schematic diagram for another perspective of Fig. 1;
  • Fig. 3 is a schematic structural view of the reaction chamber body involved in the present invention.
  • Fig. 4 is a structural schematic diagram showing only the partition of the reaction device of the first structure involved in the present invention.
  • Fig. 5 is an explanatory diagram illustrating that the internal space of the reaction chamber involved in the present invention is divided;
  • FIG. 6 is a schematic structural view of the reaction chamber body in which the internal space of the reaction device with the first structure is partially separated by partitions according to the present invention
  • Fig. 7 is a kind of structural representation of the spacer of the present invention.
  • Fig. 8 is the overall structure schematic diagram of the reaction device of the second structure involved in the present invention.
  • FIG. 9 is a schematic diagram of another viewing angle for FIG. 8;
  • Fig. 10 is an assembly schematic diagram of an air intake unit, an exhaust unit and a cover assembly related to the present invention
  • Fig. 11 is a schematic diagram of the overall structure of the reaction device of the third structure involved in the present invention.
  • Fig. 12 is a side view of Fig. 11;
  • Fig. 13 is the experimental result diagram of Experimental Example 1 involved in the present invention.
  • FIG. 14 is a graph showing the experimental results of Experimental Example 2 of the present invention.
  • the direction in which the axial length of the reaction chamber body is longer is called the “length direction” of the reaction chamber body, and vice versa, it is called the “width direction”.
  • this horizontal direction is the length direction.
  • the “lengthwise direction” referred to herein is a generally equivalent direction, rather than an absolutely consistent direction.
  • the “width direction” also refers to a substantially equivalent direction.
  • Fig. 1 is the overall structure schematic diagram of the first reaction device involved in the present invention
  • Fig. 2 is a schematic diagram for another perspective of Fig. 1;
  • Fig. 3 is a schematic structural view of the reaction chamber body involved in the present invention.
  • Fig. 4 is a structural schematic diagram showing only partitions of the reaction device of the first structure involved in the present invention.
  • the reaction device 100 for in vitro biosynthesis includes: a reaction chamber body 10 and a partition 20 .
  • the reaction chamber body 10 is used for accommodating the reaction liquid for synthesis to carry out the reaction of in vitro biosynthesis.
  • the reaction chamber body 10 is provided with an opening 10a, and the opening 10a has an internal space 11 for allowing the reaction liquid for synthesis to enter the reaction chamber body to The inlet for reaction and the outlet for discharging the reacted product.
  • the reaction solution can be directly poured into the feed port, or the reaction product can be poured out from the feed port, or can be fed and discharged in other ways, for example, the feed pipe transports the reaction liquid for synthesis from the feed port to the internal space 11, Another example is to discharge the reacted product out of the inner space 11 through a discharge pipe.
  • the opening part 10a further has a ventilation unit, and the ventilation unit has an air inlet and/or an air outlet.
  • the air inlet is used to allow external gas to enter the internal space of the reaction chamber body, so that oxygen-containing gas, such as air, can be introduced to facilitate in vitro biosynthesis reactions;
  • the exhaust port is used to allow the gas in the internal space to be discharged, so that it can The unfavorable gas generated during the reaction, such as ethanol, etc., should be discharged in time, so as not to affect the synthesis efficiency and quality.
  • one or more of the inlet, the outlet, the inlet and the exhaust port are the same opening, as shown in Figure 1, the inlet, the outlet, the inlet and the exhaust port are all are the same opening, that is, the opening portion 10a, in other examples, these four openings may be different openings, or several of them may be the same opening.
  • FIG. 5 is an explanatory diagram illustrating that the internal space of the reaction chamber involved in the present invention is divided.
  • the partition 20 is used to incompletely partition the inner space of the reaction chamber body 10 .
  • the circle in the figure represents a section of the reaction chamber body
  • the inside of the circle represents the internal space
  • the arc in the middle of the circle represents the partition.
  • “Incomplete separation” is relative to “complete separation”.
  • “Complete separation” means that the entire internal space is divided into different spaces that are not connected, that is, the spaces formed by separation are completely separated. , such as using a board to completely partition a section of the entire interior space to form two spaces.
  • the section of the entire partition completely covers a section of the interior space; while "incompletely separated ", that is, it does not form a complete partition for the internal space, as shown in the right figure in Figure 5, although the left and right edges and the lower edge of the partition touch the inner wall respectively, but the upper edge does not touch the inner wall, this incomplete separation forms Interconnected space areas are specifically shown in the right figure in Figure 5.
  • the part covered by the partition is the space area, and the spaces are connected to each other through the blank part above.
  • reaction liquid in the "flowing state" can flow from one space area to another space area, and the reaction liquid flowing through the partition 20 will be separated by the partition.
  • 20 blocking increases the mixing intensity of the reaction solution during the in vitro synthesis reaction, thereby improving the reaction efficiency.
  • the “flowing state” here means that the reaction liquid is not stationary, but at least partly moves relative to the inner wall, and the reasons for this include but are not limited to the following:
  • the first type is caused only by the movement of the reaction chamber body 10, for example, only the reaction chamber body vibrates, rotates, etc.;
  • the second type flows only by other external forces:
  • the third type, the first type and the second type both play a role, for example, when the reaction chamber body 10 rotates, the partition part 20 also rotates.
  • reaction liquid when the reaction liquid is in a flowing state, due to the blocking of the partition 20, there is another function that will increase the chance of the reaction liquid contacting the inner wall of the reaction chamber body 10, and the reaction liquid that touches the inner wall, under certain conditions, such as Gravity or being rotated, etc., have the opportunity to be diluted on the inner wall, and the diluted reaction solution has a larger surface area and can be more fully in contact with the air, thereby improving the synthesis efficiency in vitro.
  • the reaction chamber body 10 is a tank body as shown in the figure. Since the tank body is spherical or ellipsoidal, compared with other shapes, the inner wall surface area is larger, and the reaction liquid has more chances to be diluted to the inner wall. And because there is no dead angle, it is not easy to produce foam during the reaction.
  • it is a horizontal tank body, that is, when the tank body is ellipsoidal, the long axis is in the horizontal direction, which avoids deposition of the reaction liquid at the bottom and is more conducive to dilution than in the vertical direction.
  • the reaction chamber body 10 in use, is allowed to rotate to complete the reaction, so that the reaction solution can be effectively diluted on the inner wall to increase the synthesis efficiency in vitro, and the degree of dilution of the reaction solution can be controlled by The size of the volume of the reaction solution added is adjusted.
  • the horizontal central axis of the reaction chamber body 10 deviates from the horizontal state by a certain angle, that is, the reaction chamber body 10 is placed obliquely, so that when the reaction chamber body 10 rotates, the reaction liquid can be more easily diluted to more inner walls superior.
  • FIG. 6 is a schematic structural view of the reaction chamber body in which the internal space of the reaction device with the first structure is partially separated by partitions according to the present invention.
  • At least one first protrusion 10b (as shown in FIG. 6 ) is provided on the inner wall of the reaction chamber body 10, and the surface area of the inner wall can be increased by the first protrusion 10b, thereby increasing the contact with the reaction liquid. The area also increases the degree to which the reaction solution is diluted.
  • the partition 20 includes at least one partition 21, thereby incompletely partitioning the internal space 11 to form a plurality of space regions, and the plurality of partitions 21 can increase the chance of mixing the reaction liquid and Chances of being diluted to the inner wall.
  • Fig. 7 is a schematic structural view of the separator of the present invention.
  • the separator 21 is in the shape of a sheet, and like this, when the reaction liquid can contact the sheet-like surface 22 of the separator 21, it can also be diluted onto the sheet-like surface 22, thus increasing Relatively speaking, the area where the same volume of reaction solution is diluted is relatively easy to be diluted, and the contact area with the air is larger, which improves the reaction efficiency.
  • the sheet-like surface 22 here, as shown in FIG. 7 , is opposite the side edge 23 . At this time, the more the number of separators 21 is, the more the area that can be in contact with the reaction solution is, the more the diluted area of the reaction solution can be increased, and the contact area between the reaction solution and the air as a whole can be increased.
  • too many separators 21 may also cause the diluted reaction solution to contact the air for too short a time, resulting in a longer reaction time, and too many separators 21, resulting in too much overall weight, which is not enough It is beneficial to use, and the overall increase in dilution will not be too large, resulting in cost waste.
  • a plurality of partitions 21 are evenly spaced, so that the internal space 11 can be incompletely divided into the same space area, so that the degree of dilution of the reaction liquid in each space area is equivalent, that is, the reaction liquid
  • the contact area is equal, the reaction efficiency is uniform, and the overall reaction effect is better.
  • the length of the separator 21 extends in a curved line or a broken line, which can increase the chance of the reaction liquid contacting the separator 21 in the same internal space, especially when the separator is sheet-shaped, the contact area can be increased. , thereby improving the synthesis efficiency.
  • the line 21A formed between the two ends of the length of the partition 21 is between the central axis 11A of the internal space 11 of the reaction chamber body 10 in the same direction as the extending direction of the length. There is an angle a between them.
  • included angle a that is, the line 21A of the connecting line is not parallel to the central axis 11A, but has an included angle, that is, it is set obliquely to the central axis 11A.
  • the included angle a Less than or equal to 60°, preferably less than or equal to 45°, more preferably less than or equal to 20°, so that the reaction liquid in each space area formed by separation can flow from one end of the reaction chamber body 10 to the other end, so that when the separator When the end of 21 is connected with the adjacent space area, it is convenient to flow from this end to the adjacent space area, improving the mixing efficiency.
  • the included angle a should not be too large, that is, the connecting line 21a should not deviate too much from the axis, otherwise the reaction liquid will accumulate at one end of the reaction chamber body 10, which is not conducive to dilution.
  • the partition 20 is arranged on the inner wall of the reaction chamber body 10. In practice, it may be rotatably fixed on the inner wall, or may be non-rotatably fixed on the inner wall. When there are multiple partitions 21 , can be completely rotatably arranged on the inner wall, and can also be partially rotatably arranged on the inner wall.
  • the partition 21 is in the shape of a sheet, one side 23 of the sheet-shaped partition 21 faces the inner wall of the reaction chamber body 10, that is, the rotatable or non-rotatable arrangement between the side 23 and the inner wall on the inner wall.
  • the included angle is not equal to 90°.
  • the flaky surface is not completely perpendicular to the inner wall, but deviates from the vertical plane by a certain angle b, which can reduce the foam generated during the reaction, improve the reaction effect, and also Allows the reaction solution to be diluted better.
  • the length of the partition 21 extends along the length direction of the reaction chamber body 10, and extends from one end to the other end of the length direction of the reaction chamber body, that is, the length of the partition is roughly equivalent to the length of the reaction chamber body. , which can ensure a larger contact area, thereby improving the synthesis efficiency.
  • At least one flow hole (not shown in the figure) is provided on the separator 21, so as to facilitate the flow of the reaction liquid from one space area to another space area.
  • the flow holes on at least one partition 21 are evenly arranged on the partition 21, for example, there are three partitions 21, and each partition 21 is provided with a plurality of flow holes, It may be that the flow holes on any one or more of the three partitions are uniformly arranged.
  • the partition 21 when the partition 21 is arranged on the inner wall, there is at least one discontinuity between the partition and the inner wall, that is, there is at least one gap between the partition 21 and the inner wall, so that it is more convenient for the reaction liquid to flow from a space area to the inner wall. Another area of space flows.
  • at least one spacer 21 is arranged equidistantly between the gaps between the inner wall, for example, three spacers 21 are arranged on the inner wall, each spacer 21 has at least one gap between the inner wall, three The discontinuities between at least one of the partitions 21 and the inner wall are arranged equidistantly.
  • At least one second protrusion 21b is arranged on the outer surface of the separator 21.
  • the surface area of the separator 21 can be increased by the second protrusion 21b, thereby increasing The contact area with the reaction solution is increased, and the degree to which the reaction solution is diluted is increased.
  • the reaction device 100 further includes an air intake unit.
  • the aforementioned air intake allows outside air to enter the inner space 11 through the air intake unit.
  • the air intake unit 30 includes at least one air intake pipe, as shown in FIG. 1 , an air intake pipe 30a is shown in FIG. 1 , and one end of the air intake pipe faces the air inlet to deliver outside air to the inner space, That is, there is a certain distance between the intake pipe and the inner space 11 .
  • the reaction device 100 further includes an exhaust unit through which the aforementioned exhaust port allows the gas in the internal space 11 to be exhausted.
  • the exhaust unit includes at least one exhaust pipe, which can discharge the gas in the internal space 11 by communicating with the aforementioned exhaust port or extending into the internal space 11 .
  • the reaction device 100 further includes: a rotating part, the rotating part is used to rotate the reaction chamber body 10, preferably, the rotating part rotates the reaction chamber body 10 by being rotatably connected with the reaction chamber body, preferably, is connected with the reaction chamber body 10
  • the rotating mounting part located on the opposite side of the opening is rotatably connected. After being rotatably connected, the opening position of the opening is inclined upward (as shown in FIG. 1 ), so that the reaction liquid can be in contact with the inner wall and the reaction chamber body 10 during rotation.
  • Partition 20 has better contact and thinning, and even if the opening is large, there will be no reaction liquid overflow, so when the opening is used as a feed port or a discharge port, it is more convenient for feeding or discharging, and when it is used as an air inlet Or when the exhaust port is used, it can ensure a larger air exchange volume.
  • the rotating part is also provided with a rotating wheel 62
  • the reaction chamber body 10 is arranged on the rotating wheel 62
  • the rotating wheel 62 rotates with the rotation of the reaction chamber body 10, so that even the reaction chamber body 10 has a very large self-weight, and can be well supported and rotated, making it easier to realize large-scale in vitro biological reaction production.
  • the rotating wheel 62 in the figure is arranged on the frame 62 a, and the reaction chamber body 10 is arranged on the rotating wheel 62 .
  • the rotating part includes a driving unit 63 , which drives the reaction chamber body to rotate.
  • the driving unit 63 drives the reaction chamber body 10 to rotate through a rotating shaft.
  • the reaction device 100 also includes a temperature adjustment device (not shown in the figure), which is used to keep the in vitro biosynthesis reaction under a predetermined temperature condition, for example, heat preservation measures can be taken for the reaction chamber body 10 , heating or cooling, etc., preferably, the temperature adjustment device is used to adjust the temperature of the gas entering the interior of the reaction chamber body to maintain a predetermined temperature condition, for example, heating the gas entering the inner space.
  • a temperature adjustment device (not shown in the figure), which is used to keep the in vitro biosynthesis reaction under a predetermined temperature condition, for example, heat preservation measures can be taken for the reaction chamber body 10 , heating or cooling, etc.
  • the temperature adjustment device is used to adjust the temperature of the gas entering the interior of the reaction chamber body to maintain a predetermined temperature condition, for example, heating the gas entering the inner space.
  • an in vitro biosynthesis method uses the aforementioned reaction device 100 to contain the synthesis reaction solution for in vitro biosynthesis. Specifically, it will be described in conjunction with the aforementioned reaction device 100 .
  • the volume of the synthesis reaction liquid used for the in vitro biosynthesis reaction accounts for less than or equal to 30% of the volume of the reaction chamber body, which can ensure that the reaction liquid is Dilute enough to ensure reaction efficiency.
  • external gas such as air
  • the inner space which facilitates the oxygen required for the reaction.
  • the gas in the internal space is exhausted, so as to prevent the waste gas generated by the reaction from affecting the synthesis.
  • the reaction chamber body 10 of the reaction device is rotated so that the reaction liquid can be diluted on the inner wall and the partition, and can be mix.
  • the speed of rotation is less than or equal to 120r/min, so that it is not easy to generate foam and ensure the quality of synthesis.
  • Fig. 8 is the overall structure schematic diagram of the reaction device of the second structure involved in the present invention.
  • FIG. 9 is a schematic view of another viewing angle of FIG. 8 .
  • FIGS. 8 and 9 there are six partitions 21 distributed evenly at intervals, and an identical space area 21 a is formed between every two adjacent partitions 21 .
  • a recessed area 24a is provided on the other side 24 of the sheet-shaped separator 21 away from the inner wall, so that when the separator 21 is arranged on the inner wall, it is convenient for the reaction liquid to flow
  • the flow between the various spatial areas improves the overall mixing efficiency.
  • one end of the air intake pipe 30 a of the air intake unit extends into the inner space 11 from the air intake.
  • At least one air outlet hole 31 is provided on the pipe wall of the part where at least one intake duct 30a enters the interior space 11, that is, when there are multiple intake ducts 30a, each intake duct is partly Entering the inner space, among these air intake ducts 30a, at least one air outlet hole 31 is provided on the wall of the inlet part of at least one air intake duct 30a, preferably, these air outlet holes 31 are evenly distributed.
  • one end of the air intake duct 30a entering the inner space may be sealed.
  • the exhaust unit includes an exhaust duct 41, and a portion between the two ends of the intake duct of the intake duct 30a is arranged in the exhaust duct, that is, the part of the intake duct 30a is arranged in the exhaust duct.
  • One end passes through one end of the exhaust pipe 41 to enter the interior space, and the other end of the air intake pipe 30a passes through the other end of the exhaust pipe 41 to communicate with the gas source, that is, a part of the air intake pipe 30a is arranged on the exhaust pipe.
  • the air intake and the exhaust are separated, so that the air intake pipe and the exhaust pipe can be arranged at the same part of the reaction chamber body, as shown in Figure 8, the air inlet and the air outlet are both arranged at the left end.
  • the reaction device 100 further includes: a cover assembly, and the cover assembly has a cover adapted to cover the feed inlet.
  • the lid assembly further includes a sealing ring, a lid lock and a locking handle.
  • the exhaust unit further includes an exhaust hole 42 disposed on the cover 51 , and the exhaust pipe 41 communicates with the internal space through the exhaust hole 42 .
  • Fig. 10 is an assembly diagram of an air intake unit, an exhaust unit and a cover assembly related to the present invention.
  • the cover assembly 1 , the air intake unit 30 and the exhaust unit 40 are an integral structure, so that, in use, by extending one end of the air intake pipe 31 into the reaction body 10, The other end of the intake pipe 31 protrudes from the exhaust pipe 41 to connect to the gas source, and the cover body 51 provided with the exhaust hole 42 is fitted and installed on the feed port, that is, the entire configuration is completed, the structure is extraordinar, and the installation is convenient .
  • Fig. 11 is a schematic diagram of the overall structure of the reaction device of the third structure involved in the present invention.
  • FIG. 12 is a side view of FIG. 11 .
  • the reaction device 300 of this embodiment further includes a casing 70 , which is adapted to the reaction chamber body 10 and used for accommodating the reaction chamber body 10 .
  • the housing 70 has any one or a combination of the following features:
  • the housing 70 is provided with an opening and closing door 71.
  • the opening and closing door 71 is provided with a ventilation hole 71a, so that the internal space 11 of the reaction chamber body 10 can be discharged from the gas or the external gas can enter the internal space 11;
  • the outside of the housing 70 is provided with a display screen 72, that is, not inside the housing 70, but on the outside, for example, on the outer surface.
  • the display screen 72 is located outside the top 73 of the housing 70 On the surface, for example, it is embedded on the outer surface.
  • the display screen 72 for example, the display of the reaction process, status and conditions can be realized, and the touch operation can also be performed to complete the reaction and discharge of the reaction product;
  • the outer side of the housing 70 is provided with a storage platform for placing articles.
  • the storage surface of the storage platform is provided with anti-slip lines 74, and preferably, the outer surface of the upper top of the housing is the storage platform;
  • the casing is provided with any one or more of the following: air intake passage, exhaust passage, feeding passage, and discharging passage.
  • the internal space 11 of the reaction chamber body 10 that allows external gas to enter by setting an air inlet passage (not shown in the figure), for example, allows gas to enter in a manner that the air inlet passage and the air inlet are communicated through the aforementioned air inlet pipe;
  • the gas in the inner space of the reaction chamber body 10 be discharged by setting the exhaust channel, for example, the gas can be removed by connecting the exhaust port with the exhaust channel through the aforementioned exhaust channel; by setting the feed channel 11a, the synthetic reaction solution Enter the internal space 11 of the reaction chamber body 10 to perform in vitro biosynthetic reactions (as shown in FIG. 12 ); and discharge the reacted product by setting a discharge channel 11 b (as shown in FIG. 12 ).
  • FIG. 3 is a schematic structural view of the reaction chamber body involved in the present invention.
  • the bioreactor provided in this embodiment includes: a reaction chamber body 10 .
  • the reaction chamber body 10 is used for accommodating the reaction liquid for synthesis to carry out the reaction of in vitro biosynthesis.
  • the reaction chamber body 10 is provided with an opening 10a, and the opening 10a has an internal space 11 for allowing the reaction liquid for synthesis to enter the reaction chamber body to The inlet for reaction and the outlet for discharging the reacted product.
  • the reaction solution can be directly poured into the feed port, or the reaction product can be poured out from the feed port, or can be fed and discharged in other ways, for example, the feed pipe transports the reaction liquid for synthesis from the feed port to the internal space 11, Another example is to discharge the reacted product out of the inner space 11 through a discharge pipe.
  • the opening part 10a further has a ventilation unit, and the ventilation unit has an air inlet and/or an air outlet.
  • the air inlet is used to allow external gas to enter the internal space of the reaction chamber body, so that oxygen-containing gas, such as air, can be introduced to facilitate the biosynthesis reaction;
  • the exhaust port is used to allow the gas in the internal space to be discharged, so that the Gases that are unfavorable to the reaction, such as ethanol, etc., generated during the reaction, should be discharged in time to avoid affecting the synthesis efficiency and quality.
  • one or more of the inlet, the outlet, the inlet and the exhaust port are the same opening, as shown in Figure 1, the inlet, the outlet, the inlet and the exhaust port are all are the same opening, that is, the opening portion 10a, in other examples, these four openings may be different openings, or several of them may be the same opening.
  • the reaction chamber body 10 is a tank body as shown in FIG. 1 .
  • the tank body is spherical or ellipsoidal, compared with other shapes, the surface area of the inner wall is larger, and the chances of the reaction liquid being diluted to the inner wall are more. , and because there is no dead angle, it is not easy to produce foam during the reaction.
  • it is a horizontal tank body, that is, when the tank body is ellipsoidal, the long axis is in the horizontal direction, which avoids deposition of the reaction liquid at the bottom and is more conducive to dilution than in the vertical direction.
  • the tank body can be made of stainless steel.
  • the reaction chamber body 10 in use, is allowed to rotate to complete the reaction, so that the reaction solution can be effectively diluted on the inner wall to increase the efficiency of biosynthesis, and the degree of dilution of the reaction solution can be controlled by The size of the volume of the reaction solution added is adjusted.
  • the horizontal central axis of the reaction chamber body 10 deviates from the horizontal state by a certain angle, that is, the reaction chamber body 10 is placed obliquely, so that when the reaction chamber body 10 rotates, the reaction liquid can be more easily diluted to more inner walls superior.
  • At least one first protrusion is provided on the inner wall of the reaction chamber body 10, through the first protrusion, the surface area of the inner wall can be increased, thereby increasing the contact area with the reaction liquid, which also increases the reaction liquid being absorbed. degree of dilution.
  • the bioreactor further includes a rotating installation part for rotating connection with the rotating part of the rotating reaction chamber body.
  • the rotating installation part is located on the opposite side of the opening part, and after the rotating connection, the opening part is inclined upward. This can make the reaction liquid better contact and dilute with the inner wall of the reaction chamber body 10 and the partition 20 during rotation, and even if the opening is large, the reaction liquid will not overflow, so when the opening is used as a feed port or discharge When it is used as an air inlet or exhaust port, it is more convenient to feed or discharge materials, and when it is used as an air inlet or exhaust port, it can ensure a larger air exchange rate.
  • the bioreactor provided in this embodiment has a simple structure and is easy to manufacture.
  • the volume of the reaction chamber body can be greater than 10 L, even reaching tens of thousands of liters, which is convenient for expansion, and the in vitro biosynthesis reaction can be completed by rotation, which is beneficial to popularization and development. application.
  • Fig. 5 is an explanatory diagram illustrating that the internal space of the reaction chamber involved in the present invention is isolated
  • FIG. 6 is a schematic structural view of the reaction chamber body in which the internal space of the reaction device with the first structure is partially separated by partitions according to the present invention.
  • the partition 21 is used to incompletely partition the inner space of the reaction chamber body 10 involved in Embodiment 4 (as shown in FIG. 6 ). (10b in Fig. 6 refers to the first protrusion mentioned in embodiment 1)
  • the circle in the figure represents a section of the reaction chamber body
  • the inside of the circle represents the internal space
  • the arc in the middle of the circle represents the partition.
  • “Incomplete separation” is relative to “complete separation”.
  • “Complete separation” means that the entire internal space is divided into different spaces that are not connected, that is, the spaces formed by separation are completely separated. , such as using a board to completely partition a section of the entire interior space to form two spaces.
  • the section of the entire partition completely covers a section of the interior space; while "incompletely separated ", that is, it does not form a complete partition to the internal space, as shown in the right figure in Figure 5, although the left and right edges and the lower edge of the partition touch the inner wall respectively, but the upper edge does not touch the inner wall, this incomplete separation forms Interconnected space areas are specifically shown in the right figure in Figure 5.
  • the part covered by the partition is the space area, and the spaces are connected to each other through the blank part above.
  • reaction liquid in the "flowing state” can flow from one space area to another space area, and the reaction liquid flowing through the partition 21 will be separated by the partition. 21 blocks and increases the mixing intensity of the reaction solution during the biosynthesis reaction, thereby improving the reaction efficiency.
  • the “flowing state” here means that the reaction liquid is not stationary, but at least partly moves relative to the inner wall, and the reasons for this include but are not limited to the following:
  • the first type is caused only by the movement of the reaction chamber body 10, for example, only the reaction chamber body vibrates, rotates, etc.;
  • the second type flows only by other external forces:
  • the third type, the first type and the second type both play a role, for example, when the reaction chamber body 10 rotates, the partition member 21 also rotates.
  • reaction liquid when the reaction liquid is in a flowing state, due to the barrier of the separator 21, another function is to increase the chance of the reaction liquid contacting the inner wall of the reaction chamber body 10, and the reaction liquid touching the inner wall, under certain conditions, such as Gravity or being rotated, etc., have the opportunity to be diluted on the inner wall, and the diluted reaction solution has a larger surface area and can more fully contact with the air, thereby improving the biosynthesis efficiency.
  • a plurality of partitions 21 can increase the mixing opportunity of the reaction liquid and be diluted to the inner wall Opportunity.
  • Fig. 7 is a schematic structural view of a separator involved in the present invention.
  • the separator 21 is sheet-shaped, and has two opposite sheet-shaped surfaces 22 and side edges 23 disposed on both sides of the two opposite sheet-shaped surfaces.
  • the reaction solution can contact the sheet-like surface 22 of the separator 21, it can also be diluted onto the sheet-like surface 22.
  • the area where the same volume of the reaction solution is diluted is increased, and relatively speaking, it is also easy to be spread. The thinner it is, the larger the contact area with the air, which improves the reaction efficiency.
  • the sheet-like surface 22 here, as shown in FIG. 7 is opposite the side edge 23 .
  • too many separators 21 may also cause the diluted reaction solution to contact the air for too short a time, resulting in a longer reaction time, and too many separators 21, resulting in too much overall weight, which is not enough It is beneficial to use, and the overall increase in dilution will not be too large, resulting in cost waste.
  • a plurality of partitions 21 are evenly spaced, so that the internal space 11 can be incompletely divided into the same space area, so that the degree of dilution of the reaction liquid in each space area is equivalent, that is, the reaction liquid
  • the contact area is equal, the reaction efficiency is uniform, and the overall reaction effect is better.
  • the length of the separator 21 extends in a curved line or a broken line, which can increase the chance of the reaction liquid contacting the separator 21 in the same internal space, especially when the separator is sheet-shaped, the contact area can be increased. , thereby increasing the efficiency of in vitro biological reactions, for example, increasing the efficiency of in vitro protein synthesis reactions.
  • the line 21A formed between the two ends of the length of the partition 21 is between the central axis 11A of the internal space 11 of the reaction chamber body 10 in the same direction as the extending direction of the length. There is an angle a between them.
  • included angle a that is, the line 21A of the connecting line is not parallel to the central axis 11A, but has an included angle, that is, it is set obliquely to the central axis 11A.
  • the included angle a Less than or equal to 60°, preferably less than or equal to 45°, more preferably less than or equal to 20°, so that the reaction liquid in each space area formed by separation can flow from one end of the reaction chamber body 10 to the other end, so that when the separator When the end of 21 is connected with the adjacent space area, it is convenient to flow from this end to the adjacent space area, improving the mixing efficiency.
  • the included angle a should not be too large, that is, the connecting line 21a should not deviate too much from the axis, otherwise the reaction liquid will accumulate at one end of the reaction chamber body 10, which is not conducive to dilution.
  • the partition 20 is arranged on the inner wall of the reaction chamber body 10. In practice, it may be rotatably fixed on the inner wall, or may be non-rotatably fixed on the inner wall. When there are multiple partitions 21 , can be completely rotatably arranged on the inner wall, and can also be partially rotatably arranged on the inner wall.
  • the partition 21 is in the shape of a sheet, one side 23 of the sheet-shaped partition 21 faces the inner wall of the reaction chamber body 10, that is, the rotatable or non-rotatable arrangement between the side 23 and the inner wall on the inner wall.
  • the included angle is not equal to 90°.
  • the flaky surface is not completely perpendicular to the inner wall, but deviates from the vertical plane by a certain angle b, which can reduce the foam generated during the reaction, improve the reaction effect, and also Allows the reaction solution to be diluted better.
  • the length of the partition 21 extends along the length direction of the reaction chamber body 10, and extends from one end to the other end of the length direction of the reaction chamber body, that is, the length of the partition is roughly equivalent to the length of the reaction chamber body. , which can ensure a larger contact area, thereby improving the synthesis efficiency.
  • FIG. 9 shows a side sectional view of a structure in which the reaction chamber body is not completely separated by a partition.
  • a side 24 of a sheet-shaped separator 21 is provided with a recessed region 24 a, so that when the separator 21 is disposed on the inner wall, the recessed region 24 a is formed as shown in FIG. 9 .
  • the display is set away from the inner wall, which can facilitate the flow of the reaction liquid between the various space areas and improve the overall mixing efficiency.
  • At least one flow hole (not shown in the figure) is provided on the separator 21, so as to facilitate the flow of the reaction liquid from one space area to another space area.
  • the flow holes on at least one partition 21 are evenly arranged on the partition 21, for example, there are three partitions 21, and each partition 21 is provided with a plurality of flow holes, It may be that the flow holes on any one or more of the three partitions are uniformly arranged.
  • At least one second protrusion 21b is arranged on the outer surface of the separator 21.
  • the surface area of the separator 21 can be increased by the second protrusion 21b, thereby increasing The contact area with the reaction solution is increased, and the degree to which the reaction solution is diluted is increased.
  • Fig. 11 is a schematic diagram of the overall structure of the reaction device of the third structure involved in the present invention.
  • FIG. 12 is a side view of FIG. 11 .
  • the casing 70 provided in this embodiment is adapted to the reaction chamber body 10 and is used for accommodating the reaction chamber body 10 .
  • the housing 70 has any one or a combination of the following features:
  • the housing 70 is provided with an opening and closing door 71.
  • the opening and closing door 71 is provided with a ventilation hole 71a, so that the internal space 11 of the reaction chamber body 10 can be discharged from the gas or the external gas can enter the internal space 11;
  • the outside of the housing 70 is provided with a display screen 72, that is, not inside the housing 70, but on the outside, for example, on the outer surface.
  • the display screen 72 is located outside the top 73 of the housing 70 On the surface, for example, it is embedded on the outer surface.
  • the display screen 72 for example, the display of the reaction process, status and conditions can be realized, and the touch operation can also be performed to complete the reaction and discharge of the reaction product;
  • the outer side of the housing 70 is provided with a storage platform for placing articles.
  • the storage surface of the storage platform is provided with anti-slip lines 74, and preferably, the outer surface of the upper top of the housing is the storage platform;
  • the casing is provided with any one or more of the following: air intake passage, exhaust passage, feeding passage, and discharging passage.
  • the internal space 11 of the reaction chamber body 10 that allows external gas to enter by setting an air inlet passage (not shown in the figure), for example, allows gas to enter in a manner that the air inlet passage and the air inlet are communicated through the aforementioned air inlet pipe;
  • the gas in the inner space of the reaction chamber body 10 be discharged by setting the exhaust channel, for example, the gas can be removed by connecting the exhaust port with the exhaust channel through the aforementioned exhaust channel; by setting the feed channel 11a, the synthetic reaction solution Enter the internal space 11 of the reaction chamber body 10 to perform in vitro biosynthetic reactions (as shown in FIG. 12 ); and discharge the reacted product by setting a discharge channel 11 b (as shown in FIG. 12 ).
  • Fig. 1 is the overall structural representation of the reaction device of the first kind of structure that the present invention relates to;
  • FIG. 2 is a schematic diagram of another perspective of FIG. 1 .
  • the reaction device 100 for in vitro biosynthesis includes: a bioreactor and a partition 20, wherein the bioreactor is the bioreactor of Example 4, and the partition 20 has at least A partition 21 of Embodiment 5, at least one partition 21 performs the aforementioned incomplete partition on the internal space of the reaction chamber body 10 of the bioreactor.
  • the partition 21 when the partition 21 is arranged on the inner wall, there is at least one discontinuity between the partition and the inner wall, that is, there is at least one gap between the partition 21 and the inner wall, so that it is more convenient for the reaction liquid to flow from a space area to the inner wall. Another area of space flows.
  • at least one spacer 21 is arranged equidistantly between the gaps between the inner wall, for example, three spacers 21 are arranged on the inner wall, each spacer 21 has at least one gap between the inner wall, three The discontinuities between at least one of the partitions 21 and the inner wall are arranged equidistantly.
  • the reaction device 100 further includes an air intake unit.
  • the aforementioned air intake allows outside air to enter the inner space 11 through the air intake unit.
  • the air intake unit 30 includes at least one air intake pipe, as shown in FIG. 1 , an air intake pipe 30a is shown in FIG. 1 , and one end of the air intake pipe faces the air inlet to deliver outside air to the inner space, That is, there is a certain distance between the intake pipe and the inner space 11 .
  • the reaction device 100 further includes an exhaust unit through which the aforementioned exhaust port allows the gas in the internal space 11 to be exhausted.
  • the exhaust unit includes at least one exhaust pipe, which can discharge the gas in the internal space 11 by communicating with the aforementioned exhaust port or extending into the internal space 11 .
  • the reaction device 100 further includes: a rotating part, the rotating part is used to rotate the reaction chamber body 10, preferably, the rotating part rotates the reaction chamber body 10 by being rotatably connected with the reaction chamber body, preferably, is connected with the reaction chamber body 10, the rotating mounting part on the opposite side of the opening is rotatably connected, and after being rotatably connected, the opening position of the opening is inclined upward (as shown in Figure 1), so that the reaction liquid can be in contact with the inner wall of the reaction chamber body 10 during rotation.
  • the opening position of the opening is inclined upward (as shown in Figure 1), so that the reaction liquid can be in contact with the inner wall of the reaction chamber body 10 during rotation.
  • the rotating part is also provided with a wheel 62
  • the reaction chamber body 10 is arranged on the rotating wheel 62
  • the rotating wheel 62 rotates with the rotation of the reaction chamber body 10, so that even the reaction chamber body 10 has a very large self-weight, and can be well supported and rotated, making it easier to realize large-scale in vitro biological reaction production.
  • the rotating wheel 62 in the figure is arranged on the frame 62 a, and the reaction chamber body 10 is arranged on the rotating wheel 62 .
  • the rotating part includes a driving unit 63 , which drives the reaction chamber body to rotate.
  • the driving unit 63 drives the reaction chamber body 10 to rotate through a rotating shaft.
  • the reaction device 100 also includes a temperature adjustment device (not shown in the figure), which is used to keep the in vitro biosynthesis reaction under a predetermined temperature condition, for example, heat preservation measures can be taken for the reaction chamber body 10 , heating or cooling, etc., preferably, the temperature adjustment device is used to adjust the temperature of the gas entering the interior of the reaction chamber body to maintain a predetermined temperature condition, for example, heating the gas entering the inner space.
  • a temperature adjustment device (not shown in the figure), which is used to keep the in vitro biosynthesis reaction under a predetermined temperature condition, for example, heat preservation measures can be taken for the reaction chamber body 10 , heating or cooling, etc.
  • the temperature adjustment device is used to adjust the temperature of the gas entering the interior of the reaction chamber body to maintain a predetermined temperature condition, for example, heating the gas entering the inner space.
  • the reaction device of this embodiment further includes a casing adapted to the reaction chamber body 10 for accommodating the reaction chamber body, wherein the casing is the casing of the sixth embodiment.
  • an in vitro biosynthesis method uses the aforementioned reaction device 100 to contain the synthesis reaction solution for in vitro biosynthesis. Specifically, it will be described in conjunction with the aforementioned reaction device 100 .
  • the volume of the synthesis reaction liquid used for the in vitro biosynthesis reaction accounts for less than or equal to 30% of the volume of the reaction chamber body, which can ensure that the reaction liquid is Dilute enough to ensure reaction efficiency.
  • external gas such as air
  • the inner space which facilitates the oxygen required for the reaction.
  • the gas in the internal space is exhausted, so as to prevent the waste gas generated by the reaction from affecting the synthesis.
  • the reaction chamber body 10 of the reaction device is rotated so that the reaction liquid can be diluted on the inner wall and the partition, and can be mix.
  • the rotation speed is less than or equal to 120r/min, which is not easy to generate foam and ensures the quality of in vitro biosynthesis.
  • the first type of experimental device adopting the reaction device of the first type of structure, the inner wall of the reaction chamber body is provided with first protrusions, and the outer surface of the partition is equipped with second protrusions;
  • the second experimental device the difference with the first experimental device is that the inner wall of the reaction chamber body and the outer surface of the partition are smooth;
  • the samples tested for EGFP fluorescence values after 1h, 2h, 3h, 4h, 5h, and 6h of reaction were taken above, and the results are shown in Figure 13.
  • the figure shows that the reaction device provided by the present invention has a higher expression activity of the target protein than a 24-well plate , can be maintained, and has a raised reaction device, the fluorescence value of the IVTT product is higher, that is, the IVTT activity is better.
  • Figure 14 is the reaction device adopting the second structure, through the in vitro biosynthesis method of the present invention, the activity test results of the target protein EGFP prepared by reacting for 1h, 3h and 6h respectively, the volume of the reaction chamber body is 50L, and the added IVTT reaction The volume of the solution is 10L, and the control group is the target protein prepared in the laboratory using a 24-well plate. Add 300uL of the same IVTT reaction solution to the 24-well plate, and perform in vitro protein synthesis for the two at the same rotational speed and temperature conditions. biological response.
  • the reaction device for in vitro biosynthesis of the present invention using the reaction device for in vitro biosynthesis of the present invention, under three reaction times, the activity of the target protein EGFP obtained is equivalent to that of the control group, and the activity is well maintained, confirming that the in vitro biosynthesis of the present invention
  • the synthetic reaction device can carry out the scale-up production of in vitro biosynthesis.
  • both the intake and feed can be accomplished by pumping, and the exhaust and discharge can also be accomplished by suction.

Landscapes

  • Peptides Or Proteins (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

本发明提供一种体外生物合成的反应装置以及体外生物合成方法，以解决目前体外生物合成不能扩大化生产的问题，其中的体外生物合成的反应装置，其特征在于，包括：反应本体，反应本体用于容纳蛋白合成反应液以进行体外生物合成的反应，反应本体上设置有开口部，开口部具有用于让合成用反应液进入反应本体内的进料口和让反应后的产物排出的出料口；分隔部，分隔部用于对反应本体的内部空间进行不完全地分隔。

Description

体外生物合成的反应装置以及体外生物合成方法 技术领域
本发明属于生物技术领域，具体涉及一种体外生物合成的反应装置以及体外生物合成方法。
背景技术
各种体外生物合成，是相比细胞体内的合成而言，例如体外无细胞法合成蛋白质(体外蛋白质合成，体外无细胞蛋白质合成)：主要依赖于无细胞表达体系，是以外源目的DNA为蛋白质合成模板，通过人工控制补加蛋白质合成所需的底物和转录、翻译相关蛋白的辅助因子等物质，也即通过反应液实现目的蛋白质的体外合成。
然而，目前是在实验室小型完成体外合成，还没有能实现扩大化的生产装置。
亟待研制一种既能大幅放大容积，又能使反应活性得到最大限度保持的生产放大设备。
发明内容
本发明是为了解决目前体外生物合成不能扩大化生产的问题，为此，本发明提供了以下的技术方案。
本发明提供一种体外生物合成的反应装置，其特征在于，包括：反应腔本体，反应腔本体用于容纳合成用反应液以进行体外生物合成的反应，反应腔本体上设置有开口部，开口部具有用于让合成用反应液进入反应腔本体内的进料口和让反应后的产物排出的出料口；分隔部，分隔部用于对反应腔本体的内部空间进行不完全地分隔。
本发明提供的反应装置，还具有这样的特征：其中，反应腔本体为罐体，优选地，为卧式罐体，罐体为球形或椭球形。
本发明提供的反应装置，还具有这样的特征：其中，分隔部包括至少一个分隔件，优选地，分隔件呈片状。
本发明提供的反应装置，还具有这样的特征，多个分隔件均匀间隔分布。
本发明提供的反应装置，还具有这样的特征：其中，分隔件的长度呈曲线或折线延伸。
本发明提供的反应装置，还具有这样的特征：其中，分隔件的长度的两端之间形成的连线与该长度延伸方向同向的反应腔本体的内部空间的中心轴之间具有夹角a，优选地，夹角a小于等于60°，优选为小于等于45°，进一步优选为小于等于20°。
本发明提供的反应装置，还具有这样的特征：其中，至少一个分隔件可转动和/或不可 转动地固定设置在反应腔本体的内壁上。
本发明提供的反应装置，还具有这样的特征，分隔部设置在内壁上时，片状的分隔件的一个侧边面向反应腔本体的内壁。
本发明提供的反应装置，还具有这样的特征：分隔部设置在内壁上时，片状的分隔件的一个侧边面向反应腔本体的内壁，而分隔件的背离内壁的另一个侧边上设置有凹陷区域。
本发明提供的反应装置，还具有这样的特征，片状的分隔件的一个侧边面向反应腔本体的内壁时，分隔件的片状表面与相对的内壁之间的夹角不等于90°。
本发明提供的反应装置，还具有这样的特征：其中，在使用中，通过让反应腔本体转动进行反应，优选地，让反应腔本体的水平中心轴偏离水平状态。
本发明提供的反应装置，还具有这样的特征：其中，分隔件上设置有至少一个流动孔。优选地，至少一个分隔件上的流动孔均匀分布。
本发明提供的反应装置，还具有这样的特征：当分隔件设置在内壁上时，分隔件与内壁之间具有至少一个间断。优选地，至少一个分隔件对应的间断之间等距分布。
本发明提供的反应装置，还具有这样的特征：其中，开口部还具有换气单元，换气单元具有进气口和/或排气口，进气口用于让外部的气体进入反应腔本体的内部空间，排气口用于让内部空间的气体排出，优选地，进料口、出料口、进气口和排气口中的一个或多个为同一个开口。
本发明提供的反应装置，还具有这样的特征，还包括：进气单元和/或排气单元，其中，进气口通过该进气单元让外面气体进入内部空间，排气口通过该排气单元让让内部空间的气体排出排气单元。
本发明提供的反应装置，还具有这样的特征：其中，进气单元包括至少一个进气管道，进气管道的一端面向进气口而向内部空间输送外面气体，或进气管道的一端从进气口伸入内部空间，此时：一个优选地，让至少一个进气管道沿反应腔本体的水平方向延伸；再一个优选地，至少一个进气管道进入内部空间的部分的管壁上设置有至少一个出气孔，优选地，至少一个出气孔均匀分布，和/或进入内部空间的进气管道的一端为密封。
本发明提供的反应装置，还具有这样的特征：排气单元包括排气管道，优选地，进气管道两端之间有部分设置在排气管道中，进气管道的一端从排气管道的一端穿出伸入内部空间，进气管道的另一端从排气管道的另一端穿出与气源相连通。
本发明提供的反应装置，还具有这样的特征：还包括：盖体组件，盖体组件具有与进料 口适配用于盖合进料口的盖体。优选地，盖体组件还包括密封圈、罐盖锁扣和锁紧手柄。
本发明提供的反应装置，还具有这样的特征：其中，当包括盖体组件时，排气单元还包括排气孔，排气孔设置在盖体上，排气管道通过排气孔与内部空间相连通。
本发明提供的反应装置，还具有这样的特征，还包括：转动部，转动部用于转动反应腔本体，优选地，转动部通过与反应腔本体转动连接而转动反应腔本体，和/或转动部还设置有转轮，反应腔本体设置在转轮上，且转轮随反应腔本体转动而转动。
本发明提供的反应装置，还具有这样的特征：其中，转动部包括驱动单元，通过驱动单元驱动使得反应腔本体转动。
本发明提供的反应装置，还具有这样的特征，还包括：调温装置，用于让反应保持在预定的温度条件下进行；优选地，调温装置用于对进入反应腔本体内部的气体进行调温以保持预定的温度条件。
本发明提供的反应装置，还具有这样的特征，还包括：壳体，壳体与反应腔本体相适应而用于容纳反应腔本体，
优选地，壳体具有以下任意一个或多个特征的组合：
(1)壳体上设置有开闭门，优选地，开闭门上设置有通气孔；
(2)壳体的外侧设置有显示屏，优选地，显示屏位于壳体的上顶的外表面上；
(3)壳体的外侧设置有用于置放物品的置物台，优选地，置物台的置物表面设置有防滑纹路，再一优选地，壳体的上顶的外表面为置物台；
(4)壳体上设有以下中的任意一种或多种：让外部气体进入的反应腔本体的内部空间的进气通道、让反应腔本体的内部空间的气体排出的排气通道、让合成用反应液进入反应腔本体的内部空间进行体外生物合成的反应的进料通道、让反应后的产物排出的出料通道。
本发明还一种体外生物合成方法，其特征在于：采用前述的反应装置以容纳合成用反应液进行体外生物合成的反应。
本发明提供的体外生物合成方法，还具有这样的特征：其中，用于进行反应的合成用反应液的体积占反应腔本体的容积的占比小于等于30％。
本发明提供的体外生物合成方法，还具有这样的特征：其中，在反应的过程中引入外部气体到内部空间，和/或将内部空间的气体排出。
本发明提供的体外生物合成方法，还具有这样的特征：其中，在反应的过程中，让反应装置的反应腔本体进行转动，优选地，转动的速度小于等于120r/min。
本发明提供一种生物反应器，用于体外生物合成的反应，其特征在于，包括：反应腔本体，用于容纳合成用反应液以进行体外生物合成的反应，反应腔本体上设置有开口部，开口部具有进料口、出料口和换气单元，进料口用于让合成用反应液进入反应腔本体内，出料口用于让反应后得到产物从反应腔本体排出，换气单元具有进气口和/或排气口，进气口用于让外部的气体进入反应腔本体的内部空间，排气口用于让内部空间的气体排出，优选地，反应腔本体为罐体。
本发明提供的生物反应器，还具有这样的特征：其中，反应腔本体的内壁上布置有至少一个第一凸起。
本发明提供的生物反应器，还具有这样的特征：其中，进料口、出料口、进气口和排气口中的一个或多个为同一个开口。
本发明提供的生物反应器，还具有这样的特征，还包括：转动安装部，该转动安装部用于与转动反应腔本体的转动部转动连接，优选地，转动安装部位于开口部的对侧，当转动连接后，开口部倾斜向上。
本发明还提供一种用于生物反应的分隔件，其特征在于：用于对前述的生物反应器的反应腔本体的内部空间进行不完全地分隔。
本发明提供的分隔件，还具有这样的特征：其中，分隔件的长度呈曲线或折线延伸。
本发明提供的分隔件，还具有这样的特征：其中，分隔件呈片状，具有两个相对的片状表面和设置该两个相对的片状表面两侧的侧边。
本发明提供的分隔件，还具有这样的特征：其中，分隔件的一个侧边上设置有凹陷区域。
本发明提供的分隔件，还具有这样的特征：其中，分隔件上设置有至少一个流动孔。
本发明提供的分隔件，还具有这样的特征：其中，分隔件的外表面上布置有至少一个第二凸起。
本发明提供的分隔件，还具有这样的特征：其中，分隔件与生物反应器一体成型。
本发明还提供一种壳体，其特征在于：壳体与反应腔本体相适应而用于容纳反应腔本体。
本发明提供的壳体，具有以下任意一个或多个特征的组合：
(1)壳体上设置有开闭门，优选地，开闭门上设置有通气孔；
(2)壳体的外侧设置有显示屏，优选地，显示屏位于壳体的上 顶的外表面上；
(3)壳体的外侧设置有用于置放物品的置物台，优选地，置物台的置物表面设置有防滑纹路，再一优选地，壳体的上顶的外表面为置物台；
(4)壳体上设有以下中的任意一种或多种：让外部气体进入的反应腔本体的内部空间的进气通道、让反应腔本体的内部空间的气体排出的出气通道、让合成用反应液进入反应腔本体的内部空间进行体外生物合成的反应的进料通道、让反应后的产物排出的出料通道。
本发明还提供一种反应装置，其特征在于，包括：生物反应器和至少一个分隔件，其中，至少一个所述分隔件对生物反应器的反应腔本体的内部空间进行不完全的分隔，生物反应器与分隔件与一体成型，其中，所述生物反应器为前述的生物反应器，所述分隔件为前述的分隔件。
本发明提供的反应装置，还具有如下的特征，还包括：前述的壳体。
另外，本发明的体外生物合成，主要指例如体外蛋白质合成反应、mRNA体外合成反应等的体外生物合成。
发明作用与效果
本发明提供的体外生物合成的反应装置以及体外生物合成方法，至少具有以下作用与效果：
(1)通过分隔部对反应腔本体的内部空间的不完全分隔，当反应腔本体中容纳有用于体外生物合成反应的反应液时，“流动状态”的反应液能从一个空间区域向另一个空间区域流动，而流经分隔部的反应液会被分隔部阻挡而增加了体外生物合成反应过程中反应液的混合强度，从而提高反应效率，并且整体结构简单，可以采用不锈钢制造，便于进行扩大化制造和生产；
(2)反应过程中让反应腔本体转动，让反应液能有效地被摊薄在内壁上，增加体外生物合成效率；
(3)反应腔本体为罐体时，相比例如长方体等其他形状，由于内壁具有弧度，且没有死角，在反应中，不容易产生泡沫，保证了合成质量，并且，相比其他形状来说，同样的容积，具有更大的内壁表面积，能将反应液摊得更薄，更有利合成；
(4)分隔部为片状，增加了反应液接触面积，也就提高了反应液被摊薄的程度；
(5)分隔部为曲线延伸或折线延伸，在同样的内部空间，相比直线，同样增加了反应液 接触面积，有利反应液的摊薄；
(6)进气口用于让外部的气体进入反应腔本体的内部空间，这样，可以引入含氧气体，例如空气，从而有利体外生物合成反应；
(7)排气口用于让内部空间的气体排出，这样，可以将反应过程中产生的对反应不利的气体，例如乙醇等，及时排出，以免影响合成效率和质量；
附图说明
图1为本发明的涉及的第一种反应装置的整体结构示意图；
图2为针对图1的另一视角的示意图；
图3为本发明的涉及的反应腔本体的一种结构示意图；
图4为本发明的涉及的第一种结构的反应装置的只显示分隔部的结构示意图；
图5为本发明的涉及的反应腔体的内部空间被分隔的解释示意图；
图6为本发明的涉及的第一种结构的反应装置的内部空间被分隔部不完全分隔的反应腔本体的结构示意图；
图7为本发明的分隔件的一种结构示意图；
图8为本发明涉及的第二种结构的反应装置的整体结构示意图；
图9为针对图8的另一视角的示意图；
图10为本发明的涉及的一种进气单元、排气单元以及盖体组件的装配示意图；
图11为本发明的涉及的第三种结构的反应装置的整体结构示意图；
图12为图11的一个侧视图；
图13为本发明涉及的实验例1的实验结果图；
图14为本发明的实验例2的实验结果图。
具体实施方式
以下结合附图来说明本发明的具体实施方式。对于实施例中所用到的具体方法或材料，本领域技术人员可以在本发明技术思路的基础上，根据已有的技术进行常规的替换选择，而不仅限于本发明实施例的具体记载。
本文中，反应腔本体的轴线长度较长的方向称为反应腔本体的“长度方向”，反之，称“宽度方向”，例如，一个椭圆球体，长轴在水平状态时，这个水平方向就是长度方向。
本文所指的“长度方向”都是大体相当的方向，而不是绝对一致的方向，同样地，“宽度方向”也是指大体相当的方向。
实施例1
图1为本发明的涉及的第一种反应装置的整体结构示意图；
图2为针对图1的另一视角的示意图；
图3为本发明的涉及的反应腔本体的一种结构示意图；
图4为本发明的涉及的第一种结构的反应装置的只显示分隔部的结构示意图。
如图1-图4所示，本实施例提供的体外生物合成的反应装置100包括：反应腔本体10和分隔部20。
反应腔本体10用于容纳合成用反应液以进行体外生物合成的反应，反应腔本体10上设置有开口部10a，开口部10a具有用于让合成用反应液进入反应腔本体的内部空间11以进行反应的进料口和让反应后的产物排出的出料口。可以直接往进料口倾倒入反应液，或从出料口倾倒出反应产物，也可以通过其他方式进料和出料，例如进料管道从进料口向内部空间11输送合成用反应液，又例如通过出料管道将反应后的产物排出内部空间11。
在一示例中，开口部10a还具有换气单元，换气单元具有进气口和/或排气口。进气口用于让外部的气体进入反应腔本体的内部空间，这样，可以引入含氧气体，例如空气，从而有利体外生物合成反应；排气口用于让内部空间的气体排出，这样，可以将反应过程中产生的对反应不利的气体，例如乙醇等，及时排出，以免影响合成效率和质量。
在一示例中，进料口、出料口、进气口和排气口中的一个或多个为同一个开口，如图1中，进料口、出料、进气口和排气口都为同一开口，也即就是开口部10a，在其他示例中，这四个开口可以分别为不同的开口，或其中几个为同一开口。
图5为本发明的涉及的反应腔体的内部空间被分隔的解释示意图。
图5中，斜线覆盖的部分表示分隔部。
分隔部20用于对反应腔本体10的内部空间进行不完全地分隔。
结合图5进行解释，图中的圆代表反应腔本体的一个截面圆的内部代表内部空间，圆中间的弧形代表分隔部。“不完全地分隔”是相对“完全地分隔”来说，“完全地分隔”就是，将整个内部空间分隔成不连通的不同空间，也即指被分隔形成的空间之间是完全被隔断地，例如用一块板沿整个内部空间的某个截面全部隔断形成两个空间，此时，例如图5中左图，整个分隔部的截面完全覆盖了内部空间的一个截面；而“不完全地分隔”，就是不对内部空间形成全部的隔断，如图5中右图所示，分隔部的左右和下边缘虽然分别接触到内侧壁，但 上边缘未接触到内侧壁，这种不完全分隔形成了相互连通的空间区域，具体如图5中右图所示，截面上，分隔部覆盖的部分为空间区域，通过上面空白的部分，使得空间去也之间相互连通。
这样，由于是不完全地分隔，当容纳有前述的反应液时，“流动状态”的反应液能从一个空间区域向另一个空间区域流动，而流经分隔部20的反应液会被分隔部20阻挡而增加了体外合成反应过程中反应液的混合强度，从而提高反应效率。
这里的“流动状态”是指反应液是非静止不动地，而是至少部分发生了相对内壁的位置移动，导致这样的原因包括但不限于以下几种：
第一种，仅仅由于反应腔本体10运动而导致，例如仅仅是反应腔本体震动、转动等；
第二种，仅仅受其他外力而流动：
(a)例如，被外力搅动而导致，例如分隔部20转动而搅动反应液使得反应液进入流动状态；
(b)又例如，通入高压气流或超声震动引起的反应液波动使得反应液进入流动状态等。
第三种，第一种和第二种均发挥作用，例如反应腔本体10转动的同时，分隔部20也进行转动。
另外，当反应液在流动状态，由于分隔部20的阻挡，还有一个作用是，会增加反应液接触到反应腔本体10的内壁的机会，接触到内壁的反应液，在一定条件下，例如重力或被旋转等，有被摊薄在内壁上的机会，而被摊薄的反应液，具有更大的表面积，能更充分地与空气接触，从而提高了体外合成效率。
在一个示例中，反应腔本体10为如图所示的罐体，由于罐体为球形或椭球形，相比其他形状来说，内壁表面积更大，反应液被摊薄到内壁的机会更多，而且由于无死角，反应中，不容易产生泡沫。优选地，为卧式罐体，也即当罐体为椭球形时，长轴是在水平方向，相比在竖直方向，避免了反应液在底部沉积，更有利于摊薄。
在一个示例中，在使用中，让反应腔本体10进行转动以完成反应，这样，可让反应液能有效地被摊薄在内壁上，增加体外合成效率，而反应液被摊薄的程度可通过控制加入的反应液的体积的大小来调节。优选地，让反应腔本体10的水平中心轴偏离水平状态一定角度，也即反应腔本体10是倾斜放置的，这样当反应腔本体10转动时，能让反应液更容易摊薄到更多的内壁上。
图6为本发明的涉及的第一种结构的反应装置的内部空间被分隔部不完全分隔的反应腔本体的结构示意图。
在一个示例中，反应腔本体10的内壁上设置有至少一个第一凸起10b(如图6所示)，通过第一凸起10b，可以增加内壁的表面积，从而增加了与反应液的接触面积，也就增加了反应液被摊薄的程度。
在一个示例中，如图4所示，分隔部20包括至少一个分隔件21，从而将内部空间11不完全地分隔而形成多个空间区域，多个分隔件21可以增加反应液的混合机会和被摊薄到内壁的机会。
图7为本发明的分隔件的一种结构示意图。
如图7所示，在一示例中，分隔件21呈片状，这样，当反应液能接触到分隔件21的片状表面22时，也能被摊薄到片状表面22上，这样，增加了同样体积的反应液被摊薄的面积，相对来说，也容易被摊得更薄，与空气的接触面积也就更大，提高反应效率。这里的片状表面22，如图7所示，是相对侧边23来说。此时，分隔件21数量越多，与反应液能接触的面积也就越多，也就越能增加反应液的摊薄面积，进而增加了反应液整体与空气的接触面积。当然，分隔件21的数量过多，也可能会使得每次被摊薄的反应液接触空气的时间太短，导致需要更长的反应时间，并且分隔件21太多，导致整体重量太大，不利于使用，而整体上对摊薄增加不会太大，造成成本浪费。
在一示例中，多个分隔件21均匀间隔分布，这样可以将内部空间11不完全地分隔为相同的空间区域，使得每个空间区域内的反应液的摊薄程度是相当的，即反应液的接触面积相当，反应效率均匀，使整体的反应效果更好。
在一示例中，分隔件21的长度呈曲线或折线延伸，这样能在同样的内部空间内，增加反应液与分隔件21接触的机会，特别当分隔件是片状时，更能增加接触面积，从而提高合成效率。
在一示例中，如图6所示，分隔件21的长度的两端之间形成的连线所在线21A与该长度的延伸方向同向的反应腔本体10的内部空间11的中心轴11A之间具有夹角a。
这里的“与该长度的延伸方向同向…的中心轴”，具体解释就是，当分隔件21的长度是大致地沿反应腔本体10的长度方向延伸时，那么，这个中心轴11A就是这个反应腔本体10的长度方向的中心轴，当分隔件21的长度是大致地沿反应腔本体的宽度方向延伸时，那么这个中心轴11A就是这个反应腔本体10的宽度方向的中心轴，这里的同向，是指大致相 同，不是绝对相同。
这里的具有夹角a的含义，也即连线所在线21A与中心轴11A之间不是平行的，而是具有夹角的，也即倾斜于中心轴11A设置，优选地，所述夹角a小于等于60°，优选为小于等于45°，进一步优选为小于等于20°，这样可方便被分隔形成的各空间区域的反应液从反应腔本体10的一端流动至另一端，这样，当分隔件21的端部与相邻空间区域之间是连通的时，就方便从这个端部能流至相邻的空间区域，提高混合效率。夹角a不宜过大，也即连线21a不宜偏离轴线过多，否则反应液将会堆积到反应腔本体10的一端，不利于摊薄。
在一示例中，分隔部20设置在反应腔本体10的内壁上，实施中，可以是能转动地固定在内壁上，也可以是不可转动地固定在内壁上，当有多个分隔件21时，可以全部可转动地设置在内壁上，也可以部分可转动地设置在内壁上。优选地，当分隔件21是片状时，片状的分隔件21的一个侧边23面向反应腔本体10的内壁，也即该通过该侧边23与内壁之间的可转动或不可转动设置在内壁上。
在一示例中，当片状的分隔件21的一个侧边23面向反应腔本体10的内壁时，片状的分隔件的片状表面与相对(指侧边23位于的内壁)的内壁之间的夹角不等于90°，其实就是片状表面并不是完全垂直于内壁的，而是偏离垂直面一定角度b，这样相对垂直来说，可以减少反应中产生的泡沫，提高反应效果，并且也能让反应液更好地被摊薄。
在一示例中，分隔件21的长度沿反应腔本体10的长度方向延伸，且从该反应腔本体的长度方向的一端延伸到另一端，也即分隔件的长度与反应腔本体的长度大致相当，这样能保证更大的接触面积，从而提高合成效率。
在一示例中，分隔件21上设置有至少一个流动孔(图中未显示)，这样，便于反应液从一个空间区域向另一个空间区域流动。优选地，当有多个流动孔时，至少一个分隔件21上的流动孔均匀布置在分隔件21上，例如具有三个分隔件21，每个分隔件21上都设置有多个流动孔，可以是三个分隔件中的任意一个或多个上的流动孔均匀布置。
在一示例中，当分隔件21设置在内壁上时，分隔件与内壁之间具有至少一个间断，也即分隔件21与内壁之间存在至少一个间隙，这样更方便反应液从一个空间区域向另一个空间区域流动。优选地，至少一个分隔件21与内壁之间的间断之间等距排布，例如具有三个分隔件21设置在内壁上，每个分隔件21与内壁之间都具有至少一个间断，3个分隔件21中至少有一个与内壁之间的间断是等距排布的。
在一示例中，如图6和7所示，分隔件21的外表面上布置有至少一个第二凸起21b， 同样地，通过第二凸起21b，可以增加分隔件21的表面积，从而增加了与反应液的接触面积，也就增加了反应液被摊薄的程度。
在一示例中，反应装置100还包括进气单元。前述的进气口通过该进气单元让外面气体进入内部空间11。
在一示例中，进气单元30包括至少一个进气管道，如图1所示，图1中显示了一个进气管道30a，进气管道的一端面向进气口而向内部空间输送外面气体，也即进气管道与内部空间11之间具有一定距离。
在一示例中，反应装置100还包括排气单元，前述的排气口通过该排气单元让内部空间11的气体排出。
在一示例中，排气单元包括至少一个排气管道，排气管道可以通过与从前述排气口连通或者伸入内部空间11的方式，将内部空间11的气体排出。
在一示例中，反应装置100还包括：转动部，转动部用于转动反应本体10，优选地，转动部通过与反应腔本体转动连接而转动反应腔本体10，优选地，与反应腔本体10上位于开口部的对侧的转动安装部转动连接，当转动连接后，开口部的开口位置倾斜向上(如图1所示)，这样能使得反应液在转动中与反应腔本体10的内壁和分隔部20更好的接触和摊薄，并且可以即使开口较大也不会有反应液溢出，这样当开口作为进料口或排料口时，更便于进料或排料，当作为进气口或排气口时，又能保证更大的换气量。
在一示例中，如图1所示，转动部还设置有转轮62，反应腔本体10设置在转轮62上，且转轮62随反应腔本体10转动而转动，这样使得即使反应腔本体10自重非常大，也能被很好地支撑和转动，更容易实现大规模的体外生物反应生产。如图1所示，图中的转轮62设置在机架62a上，反应腔本体10设置在转轮62上。
在一示例中，转动部包括驱动单元63，通过驱动单元63驱动使得反应腔本体转动，如图2中所示，驱动单元63通过转轴驱动反应腔本体10转动。
在一示例中，反应装置100还包括调温装置(图中未显示)，调温装置用于让体外生物合成的反应保持在预定的温度条件下进行，例如可以对反应腔本体10采取保温措施、加热或降温等实现，优选地，调温装置用于对进入反应腔本体内部的气体进行调温以保持预定的温度条件，例如对进入内部空间的气体进行加热。
本实施例中，还提供一种体外生物合成方法，该体外生物合成方法采用前述的反应装置100以容纳合成用反应液进行体外生物合成的反应，具体地，结合前述的反应装置100进行 说明。
在一示例中，本实施例的体外生物合成方法中，用于进行体外生物合成的反应的合成用反应液的体积占反应腔本体的容积的占比小于等于30％，这样能保证反应液被足够地摊薄，保证反应效率。
在一示例中，本实施例的体外生物合成方法中，在体外生物合成的反应的过程中，引入外部气体到内部空间，比如空气进入，这样有利反应需要的氧气。
在一示例中，本实施例的体外生物合成方法中，在体外生物合成的反应的过程中，将内部空间的气体排出，以免反应产生的废气影响合成。
在一示例中，本实施例的体外生物合成方法中，在体外生物合成的反应的过程，让反应装置的反应腔本体10进行转动，以让反应液摊薄到内壁和分隔件上，并能被混合。另外，转动的速度小于等于120r/min，这样不容易产生泡沫，保证合成质量。
实施例2
本实施例中，与实施例1相同的零部件采用相同的图号，并省略相同的说明和解释。
图8为本发明涉及的第二种结构的反应装置的整体结构示意图；
图9为针对图8的另一视角的示意图。
如图8和9中所示，图中有6个分隔件21，均匀间隔分布，每两个相邻分隔件21之间，形成一个相同的空间区域21a。
在一示例中，如图9所示，片状的分隔件21的背离内壁的另一个侧边24上设置有凹陷区域24a，这样，当分隔件21设置在内壁上时，可方便反应液在各空间区域之间流动，提高整体的混合效率。
在一个示例中，如图8，进气单元的进气管道30a的一端从进气口伸入内部空间11。此时，一个优选地，让至少一个进气管道沿反应腔本体的水平方向X延伸，这样，可以让进入的气体尽量伸入到反应腔本体10的容易缺少有氧气体的另一端。
更优选地，如图8，至少一个进气管道30a进入内部空间11的部分的管壁上设置有至少一个出气孔31，也即当有多个进气管道30a时，各个进气管道都部分进入内部空间，这些进气管道30a中，有至少一个进气管道30a的进入的部分的管壁上设置至少一个出气孔31，优选地，这些出气孔31均匀分布。
在一示例中，当设置有出气孔时，可以让进入内部空间的进气管道30a的一端为密封。
在一示例中，排气单元包括排气管道41，而进气管道30a的进气管道的两端之间有部 分设置在排气管道中设置在排气管道中，也即进气管道30a的一端从排气管道41的一端穿出进入内部空间，进气管道30a的另一端从排气管道41的另一端穿出与气源相连通，也就是进气管道30a的一部分设置在排气管道41中，这样，将进气和排气之间隔离开，使得进气管道和排气管道可以设置在反应腔本体的同一个部位，如图8所示，进气和出气均设置在左端。
在一示例中，反应装置100还包括：盖体组件，盖体组件具有与进料口适配用于盖合进料口的盖体。
优选地，盖体组件还包括密封圈、罐盖锁扣和锁紧手柄。
在一示例中，排气单元还包括排气孔42，排气孔42设置在盖体51上，排气管道41通过排气孔42与内部空间相连通。
图10为本发明的涉及的一种进气单元、排气单元以及盖体组件的装配示意图。
在一示例中，如图10所示，盖体组件1、进气单元30以及排气单元40为一个整体结构，这样，使用中，通过将进气管道31的一端伸入反应本体10中，进气管道31的另一端从排气管道41中伸出接气源，并将设置有排气孔42的盖体51适配安装到进料口上，即完成了整个配置，结构精巧，安装方便。
实施例3
图11为本发明的涉及的第三种结构的反应装置的整体结构示意图；
图12为图11的一个侧视图。
如图11和12所示，本实施例中，与实施例1相同的零部件采用相同的图号，并省略相同的说明和解释。本实施例的反应装置300还包括壳体70，壳体70与反应腔本体10相适应而用于容纳反应腔本体10。
优选地，壳体70具有以下任意一个或多个特征的组合：
(1)壳体70上设置有开闭门71，优选地，开闭门71上设置有通气孔71a，这样可以实现反应腔本体10的内部空间11气体排出或外部的气体进入内部空间11；
(2)壳体70的外侧设置有显示屏72，也即不是在壳体70的内部，而是在外侧，例如设置在外表面，优选地，显示屏72位于壳体70的上顶73的外表面上，再例如，嵌入设置在外表面，通过该显示屏72，可以实现例如对反应过程、状态以及条件等的显示，也可以实现进行触摸操作完成反应和反应产物的排出等；
(3)壳体70的外侧设置有用于置放物品的置物台，优选地，置物台的置物表面设置有 防滑纹路74，再一优选地，壳体的上顶的外表面即为置物台；
(4)壳体上设有以下中的任意一种或多种：进气通道、排气通道、进料通道、出料通道。具体地，通过设置进气通道(图中未显示)让外部气体进入的反应腔本体10的内部空间11，例如通过前述的进气管道将进气通道和进气口连通的方式让气体进入；通过设置排气通道让反应腔本体10的内部空间的气体排出，例如通过前述的排气管道将排气口与排气通道连通的方式可排除气体；通过设置进料通道11a让合成用反应液进入反应腔本体10的内部空间11进行体外生物合成的反应(如图12所示)；通过设置出料通道11b让反应后的产物排出(如图12)。
实施例4
图3为本发明的涉及的反应腔本体的一种结构示意图。
如图3所示，本实施例提供的生物反应器包括：反应腔本体10。
反应腔本体10用于容纳合成用反应液以进行体外生物合成的反应，反应腔本体10上设置有开口部10a，开口部10a具有用于让合成用反应液进入反应腔本体的内部空间11以进行反应的进料口和让反应后的产物排出的出料口。可以直接往进料口倾倒入反应液，或从出料口倾倒出反应产物，也可以通过其他方式进料和出料，例如进料管道从进料口向内部空间11输送合成用反应液，又例如通过出料管道将反应后的产物排出内部空间11。
在一示例中，开口部10a还具有换气单元，换气单元具有进气口和/或排气口。进气口用于让外部的气体进入反应腔本体的内部空间，这样，可以引入含氧气体，例如空气，从而有利生物合成反应；排气口用于让内部空间的气体排出，这样，可以将反应过程中产生的对反应不利的气体，例如乙醇等，及时排出，以免影响合成效率和质量。
在一示例中，进料口、出料口、进气口和排气口中的一个或多个为同一个开口，如图1中，进料口、出料、进气口和排气口都为同一开口，也即就是开口部10a，在其他示例中，这四个开口可以分别为不同的开口，或其中几个为同一开口。
在一个示例中，反应腔本体10为如图1所示的罐体，由于罐体为球形或椭球形，相比其他形状来说，内壁表面积更大，反应液被摊薄到内壁的机会更多，而且由于无死角，反应中，不容易产生泡沫。优选地，为卧式罐体，也即当罐体为椭球形时，长轴是在水平方向，相比在竖直方向，避免了反应液在底部沉积，更有利于摊薄。
罐体可以采用不锈钢制备得到。
在一个示例中，在使用中，让反应腔本体10进行转动以完成反应，这样，可让反应液 能有效地被摊薄在内壁上，增加生物合成效率，而反应液被摊薄的程度可通过控制加入的反应液的体积的大小来调节。优选地，让反应腔本体10的水平中心轴偏离水平状态一定角度，也即反应腔本体10是倾斜放置的，这样当反应腔本体10转动时，能让反应液更容易摊薄到更多的内壁上。
在一个示例中，反应腔本体10的内壁上设置有至少一个第一凸起，通过第一凸起，可以增加内壁的表面积，从而增加了与反应液的接触面积，也就增加了反应液被摊薄的程度。
在一示例中，生物反应器还包括转动安装部，用于与转动反应腔本体的转动部转动连接，优选地，转动安装部位于开口部的对侧，当转动连接后，开口部倾斜向上。这样能使得反应液在转动中与反应腔本体10的内壁和分隔部20更好的接触和摊薄，并且可以即使开口较大也不会有反应液溢出，这样当开口作为进料口或排料口时，更便于进料或排料，当作为进气口或排气口时，又能保证更大的换气量。
本实施例提供的生物反应器，结构简单，容易制造，反应腔本体的容积实现大于10L以上的容积，甚至达到上万升，便于扩大化，通过转动就能完成体外生物合成反应，利于推广和应用。
实施例5
图5为本发明的涉及的反应腔体的内部空间被隔离的解释示意图；
图6为本发明的涉及的第一种结构的反应装置的内部空间被分隔部不完全分隔的反应腔本体的结构示意图。
图5中，斜线覆盖的部分表示分隔部。
分隔件21用于对实施例4涉及的反应腔本体10的内部空间进行不完全地分隔(如图6所示)。(图6中的10b所指即为实施例1中提到的第一凸起)
结合图5进行解释，图中的圆代表反应腔本体的一个截面圆的内部代表内部空间，圆中间的弧形代表分隔部。“不完全地分隔”是相对“完全地分隔”来说，“完全地分隔”就是，将整个内部空间分隔成不连通的不同空间，也即指被分隔形成的空间之间是完全被隔断地，例如用一块板沿整个内部空间的某个截面全部隔断形成两个空间，此时，例如图5中左图，整个分隔部的截面完全覆盖了内部空间的一个截面；而“不完全地分隔”，就是不对内部空间形成全部的隔断，如图5中右图所示，分隔件的左右和下边缘虽然分别接触到内侧壁，但上边缘未接触到内侧壁，这种不完全分隔形成了相互连通的空间区域，具体如图5中右图所示，截面上，分隔件覆盖的部分为空间区域，通过上面空白的部分，使得空间去也之间相互 连通。
这样，由于是不完全地分隔，当容纳有前述的反应液时，“流动状态”的反应液能从一个空间区域向另一个空间区域流动，而流经分隔件21的反应液会被分隔件21阻挡而增加了生物合成反应过程中反应液的混合强度，从而提高反应效率。
这里的“流动状态”是指反应液是非静止不动地，而是至少部分发生了相对内壁的位置移动，导致这样的原因包括但不限于以下几种：
第一种，仅仅由于反应腔本体10运动而导致，例如仅仅是反应腔本体震动、转动等；
第二种，仅仅受其他外力而流动：
(a)例如，被外力搅动而导致，例如分隔件21转动而搅动反应液使得反应液进入流动状态；
(b)又例如，通入高压气流或超声震动引起的反应液波动使得反应液进入流动状态等。
第三种，第一种和第二种均发挥作用，例如反应腔本体10转动的同时，分隔部件21也进行转动。
另外，当反应液在流动状态，由于分隔件21的阻挡，还有一个作用是，会增加反应液接触到反应腔本体10的内壁的机会，接触到内壁的反应液，在一定条件下，例如重力或被旋转等，有被摊薄在内壁上的机会，而被摊薄的反应液，具有更大的表面积，能更充分地与空气接触，从而提高了生物合成效率。
在一个示例中，如图6所示，至少一个分隔件21，从而将内部空间11不完全地分隔而形成多个空间区域，多个分隔件21可以增加反应液的混合机会和被摊薄到内壁的机会。
图7为本发明发明涉及的分隔件的一种结构示意图。
如图7所示，在一示例中，分隔件21呈片状，具有两个相对的片状表面22和设置该两个相对的片状表面两侧的侧边23。当反应液能接触到分隔件21的片状表面22时，也能被摊薄到片状表面22上，这样，增加了同样体积的反应液被摊薄的面积，相对来说，也容易被摊得更薄，与空气的接触面积也就更大，提高反应效率。这里的片状表面22，如图7所示，是相对侧边23来说。此时，分隔件21数量越多，与反应液能接触的面积也就越多，也就越能增加反应液的摊薄面积，进而增加了反应液整体与空气的接触面积。当然，分隔件21的数量过多，也可能会使得每次被摊薄的反应液接触空气的时间太短，导致需要更长的反应时间，并且分隔件21太多，导致整体重量太大，不利于使用，而整体上对摊薄增加不 会太大，造成成本浪费。
在一示例中，多个分隔件21均匀间隔分布，这样可以将内部空间11不完全地分隔为相同的空间区域，使得每个空间区域内的反应液的摊薄程度是相当的，即反应液的接触面积相当，反应效率均匀，使整体的反应效果更好。
在一示例中，分隔件21的长度呈曲线或折线延伸，这样能在同样的内部空间内，增加反应液与分隔件21接触的机会，特别当分隔件是片状时，更能增加接触面积，从而提高体外生物反应的效率，例如，提高体外蛋白质合成反应的效率。
在一示例中，如图6所示，分隔件21的长度的两端之间形成的连线所在线21A与该长度的延伸方向同向的反应腔本体10的内部空间11的中心轴11A之间具有夹角a。
这里的“与该长度的延伸方向同向…的中心轴”，具体解释就是，当分隔件21的长度是大致地沿反应腔本体10的长度方向延伸时，那么，这个中心轴11A就是这个反应腔本体10的长度方向的中心轴，当分隔件21的长度是大致地沿反应腔本体的宽度方向延伸时，那么这个中心轴11A就是这个反应腔本体10的宽度方向的中心轴，这里的同向，是指大致相同，不是绝对相同。
这里的具有夹角a的含义，也即连线所在线21A与中心轴11A之间不是平行的，而是具有夹角的，也即倾斜于中心轴11A设置，优选地，所述夹角a小于等于60°，优选为小于等于45°，进一步优选为小于等于20°，这样可方便被分隔形成的各空间区域的反应液从反应腔本体10的一端流动至另一端，这样，当分隔件21的端部与相邻空间区域之间是连通的时，就方便从这个端部能流至相邻的空间区域，提高混合效率。夹角a不宜过大，也即连线21a不宜偏离轴线过多，否则反应液将会堆积到反应腔本体10的一端，不利于摊薄。
在一示例中，分隔部20设置在反应腔本体10的内壁上，实施中，可以是能转动地固定在内壁上，也可以是不可转动地固定在内壁上，当有多个分隔件21时，可以全部可转动地设置在内壁上，也可以部分可转动地设置在内壁上。优选地，当分隔件21是片状时，片状的分隔件21的一个侧边23面向反应腔本体10的内壁，也即该通过该侧边23与内壁之间的可转动或不可转动设置在内壁上。
在一示例中，当片状的分隔件21的一个侧边23面向反应腔本体10的内壁时，片状的分隔件的片状表面与相对(指侧边23位于的内壁)的内壁之间的夹角不等于90°，其实就是片状表面并不是完全垂直于内壁的，而是偏离垂直面一定角度b，这样相对垂直来说，可以减少反应中产生的泡沫，提高反应效果，并且也能让反应液更好地被摊薄。
在一示例中，分隔件21的长度沿反应腔本体10的长度方向延伸，且从该反应腔本体的长度方向的一端延伸到另一端，也即分隔件的长度与反应腔本体的长度大致相当，这样能保证更大的接触面积，从而提高合成效率。
图9中显示了反应腔本体被分隔件不完全分隔的一种结构的侧剖图。
在一示例中，如图9所示，片状的分隔件21的一个侧边24上设置有凹陷区域24a，这样，当分隔件21设置在内壁上时，将凹陷区域24a如图9中所示背离内壁设置，可方便反应液在各空间区域之间流动，提高整体的混合效率。
在一示例中，分隔件21上设置有至少一个流动孔(图中未显示)，这样，便于反应液从一个空间区域向另一个空间区域流动。优选地，当有多个流动孔时，至少一个分隔件21上的流动孔均匀布置在分隔件21上，例如具有三个分隔件21，每个分隔件21上都设置有多个流动孔，可以是三个分隔件中的任意一个或多个上的流动孔均匀布置。
在一示例中，如图2和4所示，分隔件21的外表面上布置有至少一个第二凸起21b，同样地，通过第二凸起21b，可以增加分隔件21的表面积，从而增加了与反应液的接触面积，也就增加了反应液被摊薄的程度。
实施例6
本实施例中，与前述实施例中相同的零部件采用相同的图号，并省略相同的说明和解释。
图11为本发明的涉及的第三种结构的反应装置的整体结构示意图；
图12为图11的一个侧视图。
如图中的反应装置300所示，本实施例中提供的壳体70，与反应腔本体10相适应，而用于容纳反应腔本体10。
优选地，壳体70具有以下任意一个或多个特征的组合：
(1)壳体70上设置有开闭门71，优选地，开闭门71上设置有通气孔71a，这样可以实现反应腔本体10的内部空间11气体排出或外部的气体进入内部空间11；
(2)壳体70的外侧设置有显示屏72，也即不是在壳体70的内部，而是在外侧，例如设置在外表面，优选地，显示屏72位于壳体70的上顶73的外表面上，再例如，嵌入设置在外表面，通过该显示屏72，可以实现例如对反应过程、状态以及条件等的显示，也可以实现进行触摸操作完成反应和反应产物的排出等；
(3)壳体70的外侧设置有用于置放物品的置物台，优选地，置物台的置物表面设置有防滑纹路74，再一优选地，壳体的上顶的外表面即为置物台；
(4)壳体上设有以下中的任意一种或多种：进气通道、排气通道、进料通道、出料通道。具体地，通过设置进气通道(图中未显示)让外部气体进入的反应腔本体10的内部空间11，例如通过前述的进气管道将进气通道和进气口连通的方式让气体进入；通过设置排气通道让反应腔本体10的内部空间的气体排出，例如通过前述的排气管道将排气口与排气通道连通的方式可排除气体；通过设置进料通道11a让合成用反应液进入反应腔本体10的内部空间11进行体外生物合成的反应(如图12所示)；通过设置出料通道11b让反应后的产物排出(如图12)。
实施例7
图1为发明本发明涉及的第一种结构的反应装置的整体结构示意图；
图2为针对图1的另一视角的示意图。
如图1和图2所示，本实施例提供的体外生物合成的反应装置100包括：生物反应器和分隔部20，其中，生物反应器为实施例4的生物反应器，分隔部20具有至少一个实施例5的分隔件21，至少一个分隔件21对生物反应器的反应腔本体10的内部空间进行前述的不完全的分隔。
在一示例中，当分隔件21设置在内壁上时，分隔件与内壁之间具有至少一个间断，也即分隔件21与内壁之间存在至少一个间隙，这样更方便反应液从一个空间区域向另一个空间区域流动。优选地，至少一个分隔件21与内壁之间的间断之间等距排布，例如具有三个分隔件21设置在内壁上，每个分隔件21与内壁之间都具有至少一个间断，三个分隔件21中至少有一个与内壁之间的间断是等距排布的。
在一示例中，反应装置100还包括进气单元。前述的进气口通过该进气单元让外面气体进入内部空间11。
在一示例中，进气单元30包括至少一个进气管道，如图1所示，图1中显示了一个进气管道30a，进气管道的一端面向进气口而向内部空间输送外面气体，也即进气管道与内部空间11之间具有一定距离。
在一示例中，反应装置100还包括排气单元，前述的排气口通过该排气单元让内部空间11的气体排出。
在一示例中，排气单元包括至少一个排气管道，排气管道可以通过与从前述排气口连通或者伸入内部空间11的方式，将内部空间11的气体排出。
在一示例中，反应装置100还包括：转动部，转动部用于转动反应腔本体10，优选地， 转动部通过与反应腔本体转动连接而转动反应腔本体10，优选地，与反应腔本体10上位于开口部的对侧的转动安装部转动连接，当转动连接后，开口部的开口位置倾斜向上(如图1所示)，这样能使得反应液在转动中与反应腔本体10的内壁和分隔部20更好的接触和摊薄，并且可以即使开口较大也不会有反应液溢出，这样当开口作为进料口或排料口时，更便于进料或排料，当作为进气口或排气口时，又能保证更大的换气量。
在一示例中，如图1所示，转动部还设置有轮轮62，反应腔本体10设置在转轮62上，且转轮62随反应腔本体10转动而转动，这样使得即使反应腔本体10自重非常大，也能被很好地支撑和转动，更容易实现大规模的体外生物反应生产。如图1所示，图中的转轮62设置在机架62a上，反应腔本体10设置在转轮62上。
在一示例中，转动部包括驱动单元63，通过驱动单元63驱动使得反应腔本体转动，如图2中所示，驱动单元63通过转轴驱动反应腔本体10转动。
在一示例中，反应装置100还包括调温装置(图中未显示)，调温装置用于让体外生物合成的反应保持在预定的温度条件下进行，例如可以对反应腔本体10采取保温措施、加热或降温等实现，优选地，调温装置用于对进入反应腔本体内部的气体进行调温以保持预定的温度条件，例如对进入内部空间的气体进行加热。
在一示例中，本实施例的反应装置还包括与反应腔本体10相适应而用于容纳所述反应腔本体的壳体，其中，壳体为实施例6的壳体。
本实施例中，还提供一种体外生物合成方法，该体外生物合成方法采用前述的反应装置100以容纳合成用反应液进行体外生物合成的反应，具体地，结合前述的反应装置100进行说明。
在一示例中，本实施例的体外生物合成方法中，用于进行体外生物合成的反应的合成用反应液的体积占反应腔本体的容积的占比小于等于30％，这样能保证反应液被足够地摊薄，保证反应效率。
在一示例中，本实施例的体外生物合成方法中，在体外生物合成的反应的过程中，引入外部气体到内部空间，比如空气进入，这样有利反应需要的氧气。
在一示例中，本实施例的体外生物合成方法中，在体外生物合成的反应的过程中，将内部空间的气体排出，以免反应产生的废气影响合成。
在一示例中，本实施例的体外生物合成方法中，在体外生物合成的反应的过程，让反应装置的反应腔本体10进行转动，以让反应液摊薄到内壁和分隔件上，并能被混合。另外， 转动的速度小于等于120r/min，这样不容易产生泡沫，保证体外生物合成的质量。
实验例1
本实施例中，分别采用以下，采用体外转录翻译体系对目的蛋白EGFP进行表达测试：
(1)第一种实验装置：采用第一种结构的反应装置，反应腔本体的内壁均有布置有第一凸起，分隔件的外表面均有布置有第二凸起；
(2)第二种实验装置：与第一种实验装置的区别是反应腔本体的内壁以及分隔件的外表面均是光滑的；
(3)第三种实验装置：24孔板。
向第一种实验装置和第二种实验装置各自的反应腔本体的内部空间分别加入1L的同样的IVTT反应液(体外转录翻译体系，体外转录和翻译偶联的体系)在相同的转速和温度条件进行体外蛋白质合成的生物合成反应。
在24孔板中加入300uL的同样的IVTT反应液在同样的温度进行同样的生物反应作为对照。
以上取分别反应1h、2h、3h、4h、5h、6h后的样品测试EGFP荧光值，结果如图13所示，图中显示，本发明提供的反应装置，目标蛋白表达活性相比24孔板，能够得到保持，并且有凸起的反应装置，IVTT产物的荧光值更高，也即IVTT活性更优。
实验例2
图14为采用第二种结构的反应装置，通过本发明体外生物合成方法，分别反应1h、3h和6h制备得到的目标蛋白EGFP的活性测试结果，反应腔本体的容积为50L，加入的IVTT反应液的体积为10L，其中对照组为在实验室使用24孔板制备的目标蛋白，在24孔板中加入300uL的同样的IVTT反应液，对两种在相同的转速和温度条件进行体外蛋白质合成的生物反应。
由图14可见，采用本发明的体外生物合成的反应装置，在三种反应时间下，得到的目标蛋白EGFP的活性与对照组均相当，活性得到了很好的保持，证实本发明的体外生物合成的反应装置，能进行体外生物合成的扩大化生产。
需要说明的是，上述实施例中，进气和进料都可以通过泵送的方式完成，排气和排料也都可以通过抽吸的方式完成。

Claims (37)

  1. 一种体外生物合成的反应装置，其特征在于，包括：
    反应腔本体，所述反应腔本体用于容纳合成用反应液以进行体外生物合成的反应，所述反应腔本体上设置有开口部，所述开口部具有用于让所述合成用反应液进入所述反应腔本体内的进料口和让反应后的产物排出的出料口；
    分隔部，所述分隔部用于对所述反应腔本体的内部空间进行不完全地分隔。
  2. 根据权利要求1所述的反应装置，其特征在于：
    其中，所述反应腔本体为罐体，优选地，为卧式罐体。
  3. 根据权利要求1或2所述的反应装置，其特征在于：
    其中，所述分隔部包括至少一个分隔件，
    优选地，所述分隔件呈片状；和/或多个所述分隔件均匀间隔分布。
  4. 根据权利要求1-3任意一项所述的反应装置，其特征在于：
    其中，所述分隔件的长度呈曲线或折线延伸，和/或所述分隔件的长度的两端之间形成的连线所在线与该长度延伸方向同向的所述反应腔本体的内部空间的中心轴之间具有夹角a，优选地，优选地，所述夹角a小于等于60°，优选为小于等于45°，进一步优选为小于等于20°。
  5. 根据权利要求1-4所述的反应装置，其特征在于：
    其中，至少一个所述分隔件可转动和/或不可转动地固定设置在所述反应腔本体的内壁上，
    优选地，片状的分隔件的一个侧边面向所述反应腔本体的内壁，和/或片状的分隔件的背离所述内壁的另一个侧边上设置有凹陷区域，
    再优选地，片状的分隔件的一个侧边面向所述反应腔本体的内壁时，所述分隔件的片状表面与相对的所述内壁之间的夹角不等于90°。
  6. 根据权利要求1-5任意一项所述的反应装置，其特征在于：
    其中，在使用中，通过让所述反应腔本体转动进行所述反应，
    优选地，让所述反应腔本体的水平中心轴偏离水平状态。
  7. 根据权利要求3-7任意一项所述的反应装置，其特征在于：
    其中，所述分隔件上设置有至少一个流动孔；和/或，当所述分隔件设置在所述内壁上时，所述分隔件与所述内壁之间具有至少一个间断；优选地，至少一个所述分隔件上的所述流动孔均匀分布和/或至少一个所述分隔件对应的所述间断之间等距分布。
  8. 根据权利要求1-7任意一项所述的反应装置，其特征在于：
    其中，所述开口部还具有换气单元，所述换气单元具有进气口和/或排气口，所述进气口用于让外部的气体进入所述反应腔本体的内部空间，所述排气口用于让所述内部空间的气体排出；
    优选地，所述进料口、所述出料口、所述进气口和所述排气口中的一个或多个为同一个开口。
  9. 根据权利要求8所述的反应装置，其特征在于，还包括：
    进气单元和/或排气单元，其中，所述进气口通过该进气单元让外面气体进入所述内部空间，所述排气口通过该排气单元让所述内部空间的气体排出排气单元。
  10. 根据权利要求9所述的反应装置，其特征在于：
    其中，所述进气单元包括至少一个进气管道，所述进气管道的一端面向所述进气口而向所述内部空间输送所述外面气体，
    或所述进气管道的一端从所述进气口伸入所述内部空间，此时：一个优选地，让至少一个所述进气管道沿所述反应腔本体的水平方向延伸；再一个优选地，至少一个所述进气管道进入所述内部空间的部分的管壁上设置有至少一个出气孔，优选地，至少一个所述出气孔均匀分布，和/或进入所述内部空间的所述进气管道的一端为密封。
  11. 根据权利要求10所述的反应装置，其特征在于：
    所述排气单元包括排气管道，优选地，所述进气管道两端之间有部分设置在所述排气管道中，所述进气管道的一端从所述排气管道的一端穿出伸入所述内部空间，所述进气管道的另一端从所述排气管道的另一端穿出与气源相连通。
  12. 根据权利要求1-11任意一项所述的反应装置，其特征在于，还包括：
    盖体组件，所述盖体组件具有与进料口适配用于盖合所述进料口的盖体，
    优选地，所述盖体组件还包括密封圈、罐盖锁扣和锁紧手柄。
  13. 根据权利要求12所述的反应装置，其特征在于：
    其中，当引用所述权利要求11时，所述排气单元还包括排气孔，所述排气孔设置在所述盖体上，所述排气管道通过所述排气孔与所述内部空间相连通。
  14. 根据权利要求1-13任意一项所述的反应装置，其特征在于，还包括：
    转动部，所述转动部用于转动所述反应腔本体，
    优选地，所述转动部通过与所述反应腔本体转动连接而转动所述反应腔本体；
    和/或，所述转动部还设置有转轮，所述反应腔本体设置在所述转轮上，且所述转轮随所述反应腔本体转动而转动。
  15. 根据权利要求14所述的反应装置，其特征在于：
    其中，所述转动部包括驱动单元，通过所述驱动单元驱动使得所述反应腔本体转动。
  16. 根据权利要求1-15任意一项所述的反应装置，其特征在于，还包括：
    调温装置，用于让所述反应保持在预定的温度条件下进行；
    优选地，所述调温装置用于对进入所述反应腔本体内部的气体进行调温以保持所述预定的温度条件。
  17. 根据权利要求1-16任意一项所述的反应装置，其特征在于：
    其中，所述反应腔本体的内壁上布置有至少一个第一凸起，
    和/或所述分隔件的外表面上布置有至少一个第二凸起。
  18. 根据权利要求1-17任意一项所述的反应装置，其特征在于，还包括：
    壳体，所述壳体与所述反应腔本体相适应而用于容纳所述反应本体，
    优选地，所述壳体具有以下任意一个或多个特征的组合：
    所述壳体上设置有开闭门，优选地，所述开闭门上设置有通气孔；
    所述壳体的外侧设置有显示屏，优选地，所述显示屏位于所述壳体的上顶的外表面上；
    所述壳体的外侧设置有用于置放物品的置物台，优选地，所述置物台的置物表面设置有防滑纹路，再一优选地，所述壳体的上顶的外表面为所述置物台；
    所述壳体上设有以下中的任意一种或多种：让外部气体进入的所述反应腔本体的内部空间的进气通道、让所述反应腔本体的内部空间的气体排出的排气通道、让合成用反应液进入所述反应腔本体的内部空间进行体外生物合成的反应的进料通道、让反应后的产物排出的出料通道。
  19. 一种体外生物合成方法，其特征在于：
    采用权利要求1-18任意一项所述的反应装置以容纳合成用反应液进行体外生物合成的反应。
  20. 根据权利要求19所述的体外生物合成方法，其特征在于：
    其中，用于进行所述反应的合成用反应液的体积占反应腔本体的容积的比例小于等于30％。
  21. 根据权利要求19或20所述的体外生物合成方法，其特征在于：
    其中，在所述反应的过程中引入外部气体到内部空间，和/或将所述内部空间的气体排出。
  22. 根据权利要求19-21任意一项所述的体外生物合成方法，其特征在于：
    其中，在所述反应的过程中，让所述反应装置的反应腔本体进行转动，优选地，让所述反应腔本体的水平中心轴偏离水平状态；
    另一优选地，转动的速度小于等于120r/min。
  23. 一种生物反应器，用于体外生物合成的反应，其特征在于，包括：
    反应腔本体，用于容纳合成用反应液以进行体外蛋白合成的反应，所述反应腔本体上设置有开口部，所述开口部具有进料口、出料口和换气单元，所述进料口用于让所述合成用反应液进入所述反应腔本体内，所述出料口用于让反应后得到产物从所述反应腔本体排出，所述换气单元具有进气口和/或排气口，所述进气口用于让外部的气体进入所述反应腔本体的内部空间，所述排气口用于让所述内部空间的气体排出。
  24. 根据权利要求23所述的生物反应器，其特征在于：
    其中，所述反应腔本体的内壁上布置有至少一个第一凸起。
  25. 根据权利要求23或24所述的生物反应器，其特征在于：
    其中，所述进料口、所述出料口、所述进气口和所述排气口中的一个或多个为同一个开口。
  26. 根据权利要求25所述的生物反应器，其特征在于，还包括：
    转动安装部，该转动安装部用于与转动所述反应腔本体的转动部转动连接，优选地，所述转动安装部位于所述开口部的对侧，当转动连接后，所述开口部倾斜向上。
  27. 根据权利要求23-25任意一项所述的生物反应器，其特征在于：
    其中，所述反应腔本体为罐体。
  28. 一种用于生物反应的分隔件，其特征在于：
    用于对23-27任意一项所述的生物反应器的反应腔本体的内部空间进行不完全地分隔。
  29. 根据权利要求28所述的分隔件，其特征在于：
    其中，所述分隔件的长度呈曲线或折线延伸。
  30. 根据权利要求28或29所述的分隔件，其特征在于：
    其中，所述分隔件呈片状，具有两个相对的片状表面和设置该两个相对的片状表面两侧的侧边。
  31. 根据权利要求30所述的分隔件，其特征在于：
    其中，所述分隔件的一个侧边上设置有凹陷区域。
  32. 根据权利要求28-31任意一项所述的分隔件，其特征在于：
    其中，所述分隔件上设置有至少一个流动孔。
  33. 根据权利要求28-32任意一项所述的分隔件，其特征在于：
    其中，所述分隔件的外表面上布置有至少一个第二凸起。
  34. 一种壳体，其特征在于：
    所述壳体与生物反应器的反应腔本体相适应而用于容纳所述反应腔本体，其中，所述生物反应器为权利要求23-27任意一项所述的生物反应器。
  35. 根据权利要求34所述的壳体，具有以下任意一个或多个特征的组合：
    所述壳体上设置有开闭门，优选地，所述开闭门上设置有通气孔；
    所述壳体的外侧设置有显示屏，优选地，所述显示屏位于所述壳体的上顶的外表面上；
    所述壳体的外侧设置有用于置放物品的置物台，优选地，所述置物台的置物表面设置有防滑纹路，再一优选地，所述壳体的上顶的外表面为所述置物台；
    所述壳体上设有以下中的任意一种或多种：让外部气体进入的所述反应腔本体的内部空间的进气通道、让所述反应腔本体的内部空间的气体排出的出气通道、让合成用反应液进入所述反应腔本体的内部空间进行体外生物合成的反应的进料通道、让反应后的产物排出的出料通道。
  36. 一种反应装置，其特征在于，包括：
    生物反应器和至少一个分隔件，其中，至少一个所述分隔件对生物反应器的反应腔本体的内部空间进行不完全的分隔，生物反应器与分隔件与一体成型，其中，所述生物反应器为权利要求23-27任意一项所述的生物反应器，所述分隔件为权利要求28-33任意一项所述的分隔件。
  37. 根据权利要求36所述的反应装置，其特征在于，还包括：
    权利要求34或35所述的壳体。
PCT/CN2022/135274 2021-11-30 2022-11-30 体外生物合成的反应装置以及体外生物合成方法 WO2023098697A2 (zh)

Applications Claiming Priority (12)

Application Number Priority Date Filing Date Title
CN202111448968 2021-11-30
CN202111448968.7 2021-11-30
CN202210095041.8 2022-01-26
CN202210095041.8A CN114307940A (zh) 2021-11-30 2022-01-26 一种卧式自搅拌体外蛋白合成反应器的罐体装置
CN202210093587.XA CN114247415A (zh) 2021-11-30 2022-01-26 一种卧式自搅拌体外蛋白合成反应器及体外蛋白合成方法
CN202210093573.8 2022-01-26
CN202210093573.8A CN114471367A (zh) 2021-11-30 2022-01-26 一种卧式自搅拌体外蛋白合成反应器的罐盖集成式进排气装置
CN202210093587.X 2022-01-26
CN202220528915.XU CN219279864U (zh) 2021-11-30 2022-03-11 生物反应器以及相应的分隔件、反应装置和壳体
CN202210239438.X 2022-03-11
CN202220528915.X 2022-03-11
CN202210239438.XA CN114405455A (zh) 2021-11-30 2022-03-11 体外生物合成的反应装置以及体外生物合成方法

Publications (2)

Publication Number Publication Date
WO2023098697A2 true WO2023098697A2 (zh) 2023-06-08
WO2023098697A3 WO2023098697A3 (zh) 2023-08-17

Family

ID=86613191

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2022/135274 WO2023098697A2 (zh) 2021-11-30 2022-11-30 体外生物合成的反应装置以及体外生物合成方法

Country Status (1)

Country Link
WO (1) WO2023098697A2 (zh)

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2550173A (en) * 2016-05-11 2017-11-15 Perkins Engines Co Ltd Mixer for after-treatment system
CN106902663A (zh) * 2017-04-01 2017-06-30 四川群青新材料科技有限公司 流化式高效混匀静态混合器
CN207520990U (zh) * 2017-09-22 2018-06-22 地龙力生物工程（天津）有限公司 一种地龙蛋白多肽缩水合成装置
DE102017010229A1 (de) * 2017-11-06 2019-05-09 Michael Niederbacher Pfropfenstrom-Fermenter einer Biogasanlage
CN108636333A (zh) * 2018-05-18 2018-10-12 苏州市金翔钛设备有限公司 一种混料氧化反应设备
CN212335181U (zh) * 2020-04-30 2021-01-12 天津二商迎宾肉类食品有限公司 一种生物发酵合成一体反应装置
CN114247415A (zh) * 2021-11-30 2022-03-29 康码（上海）生物科技有限公司 一种卧式自搅拌体外蛋白合成反应器及体外蛋白合成方法

Also Published As

Publication number Publication date
WO2023098697A3 (zh) 2023-08-17

Similar Documents

Publication Publication Date Title
CN219279864U (zh) 生物反应器以及相应的分隔件、反应装置和壳体
CN103007865B (zh) 搅拌釜
JP5648986B2 (ja) 流体処理装置及び流体処理方法
US20190211297A1 (en) Large circulating fluidized bed cell bioreactor and method for culturing animal cells
US20230348834A1 (en) Fixed bed bioreactor vessel and methods of using the same
WO2023098697A2 (zh) 体外生物合成的反应装置以及体外生物合成方法
KR20200078221A (ko) 회분식 반응기
CN206902142U (zh) 一种复合微生物发酵装置
KR101817770B1 (ko) 고효율 유체 혼합 또는 반응 장치
CN218951490U (zh) 一种原子层沉积反应装置
CN106732290A (zh) 一种连续生产硝基甲烷的反应器
CN209057688U (zh) 一种食用菌生产用物料混合发酵装置
CN109763168A (zh) 一种单晶碳晶纳米镀膜方法及反应器
CN210545081U (zh) 一种微孔反应釜
CN212524038U (zh) 一种新型一体式气液连续流反应器
CN211725506U (zh) 一种涂料生产加工用原料混合搅拌装置
CN212417800U (zh) 搅拌槽设备改良结构
JP2002045173A (ja) 細胞塊の培養方法およびバイオリアクター
CN217454534U (zh) 塑料颗粒冷却均化装置
CN216856427U (zh) 一种化工物料玻璃搅拌釜
CN207193264U (zh) 发酵用搅拌罐
CN216663041U (zh) 密封发酵装置
CN216062092U (zh) 一种诱导冷却结晶器
CN213132790U (zh) 一种带有废气处理的化工用搅拌机
CN212357229U (zh) 发酵罐