Apparatus and method for catalytic production of phosphatidylserine

Abstract

This invention relates to the field of chemical preparation, and more particularly to an apparatus and method for the catalytic preparation of phosphatidylserine. The apparatus includes a fixed base, a reaction vessel fixedly mounted on the fixed base, a stirring device disposed within the reaction vessel, and a heating device disposed within the reaction vessel. The reaction vessel is provided with a first inlet, a second inlet, and an outlet. An observation window is provided on the peripheral side of the reaction vessel, extending into the inner cavity of the reaction vessel and covered by tempered glass. A first-type temperature sensor is provided within the inner cavity of the reaction vessel. The stirring device includes a central stirring device and an edge stirring device. The central stirring device is driven by a rotary motor and simultaneously drives the edge stirring device to mix the materials. This invention solves the technical problem of low conversion rates in existing catalytic preparation methods of phosphatidylserine due to incomplete mixing of raw materials and catalyst during the catalytic process.

Description

Technical Field

[0001] This invention relates to the field of chemical preparation, and more particularly to an apparatus and method for catalytically preparing phosphatidylserine. Background Technology

[0002] Phosphatidylserine is an important component of cell membranes, especially abundant in brain cells, where it can improve nerve cell function, enhance memory, and relieve stress. The human body can synthesize it, but the amount decreases with age, requiring acquisition through food or supplements. The main methods for preparing phosphatidylserine include enzymatic reactions, chemical synthesis, liposome preparation technology, and molecular imprinting purification. Among these, enzymatic reactions are the preferred method for industrial preparation. Enzymatic methods utilize phospholipase D to catalyze the phosphate transfer reaction between lecithin and serine, efficiently synthesizing phosphatidylserine under mild conditions. This method offers advantages such as high selectivity, environmental friendliness, and cost-effectiveness.

[0003] In the phosphotransferase reaction between lecithin and serine catalyzed by phospholipase D, the catalytic process is particularly crucial. This process requires a pH of 6.5-7.5 and a temperature of 37.5°C, allowing phospholipase D to catalyze the replacement of the choline group in phosphatidylcholine with serine, generating phosphatidylserine. During this process, it is essential to ensure uniform mixing of the materials for complete reaction, and to maintain precise and constant temperature and pH; otherwise, the conversion rate will be significantly reduced.

[0004] Chinese patent application number 202410632769.9 discloses a method for preparing phosphatidylserine. This invention optimizes specific reaction conditions and utilizes phospholipase B to convert phosphatidylcholine into phosphatidylserine, which helps reduce production costs and improve conversion rates. The invention employs a method of first heating and then rapidly cooling to promote the rapid precipitation and aggregation of phosphatidylserine, thus improving the yield of the final product. In the purification process of phosphatidylserine, the invention uses organic solvent extraction combined with ethanol slurrying, which is simple to operate, effectively reduces residual organic solvents, and allows for flexible control of solvent residues. The preparation method is simplified, and both the reaction solution and solvent can be recycled, further reducing production costs and being environmentally friendly and pollution-free, which is conducive to its widespread application in production practice. However, the technical solution proposed in the above invention does not address how to ensure uniform mixing of raw materials and catalysts to maximize conversion rates during the catalytic process, thus resulting in a lack of guaranteed conversion rates during the catalytic process. Summary of the Invention

[0005] Therefore, in view of the above problems, the present invention proposes an apparatus and preparation method for the catalytic preparation of phosphatidylserine, which solves the technical problem that the conversion rate is low in the existing catalytic preparation of phosphatidylserine due to incomplete mixing of raw materials and catalyst in the catalytic process.

[0006] To achieve the above objectives, the present invention adopts the following technical solution: an apparatus for catalytic preparation of phosphatidylserine, comprising a fixed base, a reaction vessel fixedly mounted on the fixed base, a stirring device disposed within the reaction vessel, and a heating device disposed within the reaction vessel. The reaction vessel is provided with a first inlet, a second inlet, and an outlet. An observation window is provided on the peripheral side of the reaction vessel, extending into the inner cavity of the reaction vessel and covered by tempered glass. A first-type temperature sensor is provided within the inner cavity of the reaction vessel. The stirring device includes a central stirring device and an edge stirring device. The central stirring device is driven by a rotary motor and simultaneously drives the edge stirring device to mix and stir the materials. The central stirring device is located at the central axis of the reaction vessel, and the edge stirring device is located at the inner side wall of the reaction vessel. The central stirring device and the edge stirring device work in conjunction to perform stirring operations.

[0007] Furthermore, the central stirring device includes a rotary motor, a rotating shaft fixedly connected to the drive end of the rotary motor, and a stirring ring assembly disposed on the rotating shaft. The stirring ring assembly includes several central support rods fixedly connected to the rotating shaft and a central stirring ring fitted onto each of the central support rods. The central support rods are inclined to the horizontal plane, and the angle between them and the rotating shaft is 100°-120°. The central stirring ring is fixedly connected to the free end of each of the central support rods. A first top block is disposed on one side of the central stirring ring near the inner cavity of the reactor.

[0008] Furthermore, the edge stirring device includes an annular locking groove formed on the inner side of the reactor, an annular functional plate locked in the annular locking groove, and a functional component disposed on the annular functional plate. The annular functional plate has an annular functional cavity, and the functional component is disposed in the annular functional cavity. A plurality of first connecting grooves are arranged around the side of the annular functional plate near the rotating shaft, and each first connecting groove extends into the annular functional cavity. The upper end face and the lower end face of the annular functional plate are respectively provided with a second connecting groove and a third connecting groove. The functional component extends outward through the first connecting groove, the second connecting groove and the third connecting groove, and the functional component is driven to work by the first top block.

[0009] Furthermore, the functional ornament is provided with a rotating shaft, which is rotatably disposed in the annular functional cavity. The functional ornament includes an upper swing area, a central rotation area, and a lower swing area. A torsion spring is provided on the functional ornament, which controls the movement stroke of the functional ornament and resets it.

[0010] Furthermore, the upper swing area of ​​the functional ornament extends outward through the second connecting groove, and a second top block is provided on the central rotating area of ​​the functional ornament. The second top block extends outward through the first connecting groove, and the lower swing area of ​​the functional ornament extends outward through the third connecting groove. The diameter of the opening of the second connecting groove is greater than the width of the upper swing area, and the diameter of the opening of the third connecting groove is greater than the width of the lower swing area.

[0011] Furthermore, in the initial state, the central rotation area is set perpendicular to the horizontal plane, the upper swing area has an angle of 45°-60° with the horizontal plane, and the lower swing area has an angle of 65°-85° with the horizontal plane. The functional ornament is rotated and swung by the first top block pressing against the second top block.

[0012] Furthermore, when the first top block and the second top block come into contact with each other, the first top block pushes the second top block inward, causing the rotatable functional ornament to rotate. The contact between the first top block and the second top block is intermittent, causing the functional ornament to rotate intermittently. The upper swing area rotates away from the rotation axis, and the lower swing area rotates towards the rotation axis.

[0013] Furthermore, a second type of temperature sensor is provided at the free end of both the upper swing region and the free end of the lower swing region.

[0014] A method for catalytically preparing phosphatidylserine includes the following steps:

[0015] S1. Raw material preparation: Phospholipase D is prepared by microbial fermentation using soybean lecithin and L-serine as raw materials.

[0016] S2. The apparatus for catalytic preparation of phosphatidylserine operates at pH 6.5-7.5 and 37℃. Phospholipase D catalyzes the substitution of the choline group in phosphatidylcholine with serine to generate phosphatidylserine, with simultaneous stirring during the conversion process.

[0017] S3. Separation was performed using column chromatography, and purification was achieved by adjusting the proportion of n-hexane as the mobile phase.

[0018] Furthermore, in step S1, the microbial fermentation uses Streptomyces cinnamon, and in step S2, the catalytic reaction time is 6-8 hours. During the stirring process, the temperature of the mixture in different areas of the reactor is monitored by the second type of temperature sensor set on the functional component.

[0019] By adopting the aforementioned technical solution, the beneficial effects of the present invention are:

[0020] 1. Compared with existing catalytic chemical reagent preparation methods, this invention, especially for catalytic reaction reagents with high requirements for temperature and mixing, delivers superior performance. Furthermore, due to its uniform mixing and higher temperature sensing accuracy, it achieves a higher catalytic conversion rate. The specific structural improvement lies in the addition of an improved stirring device to enhance the uniformity of material mixing, ensuring thorough mixing and complete reaction of all chemical reagents during the re-catalytic process. The stirring device includes a central stirring device and an edge stirring device, which work together to ensure uniform material mixing. The central stirring device is responsible for mixing the material in the center of the reactor. Simultaneously, its central support rod extends inclined towards the reactor, reducing stirring resistance compared to a vertical rotation axis design and preventing deformation of the central support rod due to excessive pressure. To reinforce the central support rod during stirring, a central stirring ring is provided around it, connecting the individual central support rods to form a unified whole with higher strength. The central stirring ring serves two purposes: enhancing the strength of the central stirring device and acting as an intermediary for cooperation with the edge stirring device.

[0021] 2. The edge stirring device in this invention serves as an auxiliary to the central stirring device. It enhances the stirring and mixing of materials at the edge of the reactor, ensuring uniformity. A second type of temperature sensor is installed above it. The cooperation between the first and second type of temperature sensors enhances real-time monitoring of the temperature in all areas of the reactor, ensuring the catalytic conversion rate of the materials. This is crucial because precise temperature and pH values ​​are essential for the catalytic preparation of phosphatidylserine; otherwise, the catalytic conversion rate will be directly affected. The edge stirring device in this invention primarily utilizes an annular functional plate on the inner wall of the reactor and a functional ornament mounted above it. The rotation of the ornament stirs the mixture. The ornament itself does not have a drive source, thus saving energy. Its operation relies mainly on the central stirring device for external drive. Specifically, the first top block of the central stirring device intermittently abuts against the second top block of the edge stirring device, controlling the intermittent rotation of the ornament. The rotation and reset of the ornament are determined by… This is achieved through a torsion spring. During rotation, the upper and lower swing zones of the functional ornament rotate in opposite directions vertically, moving towards or away from the central stirring device. The ornament stirs the mixture at the edge of the reactor, working in sync with the central stirring device to ensure a more thorough and complete catalytic reaction, thus improving the catalytic conversion rate. Second-type temperature sensors are installed on both the upper and lower swing zones, working in conjunction with a first-type temperature sensor on the inner side of the reactor to monitor the temperature in various areas. Because the ornament rotates vertically, and the second-type temperature sensors are located at the free ends of the upper and lower swing zones, their horizontal height changes as the ornament rotates. This allows for temperature monitoring of the mixture at different heights within the reactor, combined with the first-type temperature sensor for more precise temperature monitoring.

[0022] 3. In the preparation method proposed in this invention, by strictly controlling the catalytic reaction conditions, temperature and pH value in step S2, and simultaneously carrying out central main stirring and edge auxiliary stirring in the reaction vessel, the conversion rate of catalytic preparation of phosphatidylserine is greatly improved. Attached Figure Description

[0023] Other features, objects, and advantages of the present invention will become more apparent from the following detailed description of non-limiting embodiments with reference to the accompanying drawings:

[0024] Figure 1 This is a front view of the overall structure of the present invention;

[0025] Figure 2 This is a schematic diagram of the three-dimensional structure of the reaction vessel of the present invention;

[0026] Figure 3 This is a schematic diagram of the internal structure of the reactor of the present invention;

[0027] Figure 4 for Figure 3 Enlarged view of point A in the middle;

[0028] Figure 5 This is a top view of the central stirring device of the present invention;

[0029] Figure 6 This is a schematic diagram of the functional ornament structure of the present invention;

[0030] Figure 7 This is a flowchart of the preparation method of the present invention;

[0031] Reference numerals: 1. Fixed base; 2. Reactor; 21. First feed inlet; 22. Second feed inlet; 23. Discharge outlet; 24. Observation window; 25. Inner cavity; 26. Type I temperature sensor; 3. Central stirring device; 31. Rotary motor; 32. Rotating shaft; 33. Stirring ring assembly; 331. Central support rod; 332. Central stirring ring; 3321. First top block; 4. Edge stirring device; 41. Annular locking groove; 42. Annular functional plate; 421. Annular functional cavity; 422. Second connecting groove; 423. First connecting groove; 424. Third connecting groove; 43. Functional ornament; 431. Upper swing area; 432. Central rotation area; 4321. Second top block; 4322. Rotating shaft; 433. Lower swing area; 5. Heating device; 6. Type II temperature sensor. Detailed Implementation

[0032] To make the technical means, creative features, objectives and effects of this invention easier to understand, the invention will be further described below in conjunction with specific embodiments.

[0033] Please see Figures 1-7This invention provides an apparatus for the catalytic preparation of phosphatidylserine, comprising a fixed base 1, a reaction vessel 2 fixedly mounted on the fixed base 1, a stirring device disposed within the reaction vessel 2, and a heating device 5 disposed within the reaction vessel 2. The reaction vessel 2 is provided with a first inlet 21, a second inlet 22, and an outlet 23. An observation window 24 is provided on the peripheral side of the reaction vessel 2, extending into the inner cavity 25 of the reaction vessel 2 and covered by tempered glass. A first-type temperature sensor 26 is provided in the inner cavity 25 of the reaction vessel 2. The stirring device includes a central stirring device 3 and an edge stirring device 4. The central stirring device 3 is driven by a rotary motor 31 and simultaneously drives the edge stirring device 4 to mix and stir the materials. The central stirring device 3 is located at the central axis of the reaction vessel 2, and the edge stirring device 4 is located at the inner side wall of the reaction vessel 2. The central stirring device 3 and the edge stirring device 4 work together to stir.

[0034] The central stirring device 3 includes a rotary motor 31, a rotating shaft 32 fixedly connected to the drive end of the rotary motor 31, and a stirring ring assembly 33 disposed on the rotating shaft 32. The stirring ring assembly 33 includes six central support rods 331 fixedly connected to the rotating shaft 32, and a central stirring ring 332 fitted onto each of the central support rods 331. The central support rods 331 are inclined to the horizontal plane, and the angle between them and the rotating shaft 32 is 110°. The central stirring ring 332 is fixedly connected to the free end of each of the central support rods 331. A first top block 3321 is provided on the side of the central stirring ring 332 near the inner cavity 25 of the reactor 2; the edge stirring device 4 includes an annular locking groove 41 formed on the inner side of the reactor 2, an annular functional plate 42 locked in the annular locking groove 41, and a functional ornament 43 disposed on the annular functional plate 42. An annular functional cavity 421 is provided in the annular functional plate 42, and the functional ornament 43 is disposed in the annular functional cavity 421. A ring is arranged around the side of the annular functional plate 42 near the rotating shaft 32. Six first connecting slots 423 extend into the annular functional cavity 421. The upper and lower surfaces of the annular functional plate 42 are respectively provided with second connecting slots 422 and third connecting slots 424. The functional ornament 43 extends outward through the first connecting slots 423, second connecting slots 422, and third connecting slots 424. The functional ornament 43 is driven by the first top block 3321. The functional ornament 43 is rotatably mounted within the annular functional cavity 421 via a rotating shaft 4322. 3 includes an upper swing area 431, a central rotation area 432, and a lower swing area 433. A torsion spring is provided on the functional swing component 43, controlling its movement and resetting. The torsion spring is built into the functional swing component 43 and connected to a rotating shaft 4322. The functional swing component 43 rotates vertically via the rotating shaft 4322. When the upper swing area 431 approaches the rotating shaft 32, the lower swing area 433 moves away from the rotating shaft 32; when the upper swing area 431 moves away from the rotating shaft 32, the lower swing area 433 approaches the rotating shaft 32. See [reference needed for rotation state] for details. Figure 6 .

[0035] The upper swing area 431 of the functional ornament 43 extends outward through the second connecting groove 422. A second top block 4321 is provided on the central rotating area 432 of the functional ornament 43. The second top block 4321 extends outward through the first connecting groove 423. The lower swing area 433 of the functional ornament 43 extends outward through the third connecting groove 424. The diameter of the opening of the second connecting groove 422 is greater than the width of the upper swing area 431, and the diameter of the opening of the third connecting groove 424 is greater than the width of the lower swing area 433.

[0036] In its initial state, the central rotation area 432 is perpendicular to the horizontal plane. The upper swing area 431 forms a 45° angle with the horizontal plane, and the lower swing area 433 forms an 80° angle with the horizontal plane. The functional swing component 43 rotates and swings by the first top block 3321 pressing against the second top block 4321. When the first top block 3321 and the second top block 4321 contact each other, the first top block 3321 pushes the second top block 4321 inward, causing the rotatable functional swing component 43 to rotate. The contact between the first top block 3321 and the second top block 4321 is intermittent, causing the functional swing component 43 to rotate intermittently. The upper swing area 431 rotates away from the rotation axis 32, and the lower swing area 433 rotates towards the rotation axis 32. A second type of temperature sensor 6 is provided at the free end of both the upper swing area 431 and the lower swing area 433.

[0037] This embodiment also proposes a method for the catalytic preparation of phosphatidylserine, which involves the above-mentioned apparatus and includes the following steps:

[0038] S1. Raw material preparation: Phospholipase D is prepared by microbial fermentation using soybean lecithin and L-serine as raw materials.

[0039] S2. The apparatus for catalytic preparation of phosphatidylserine operates at pH 6.5-7.5 and 37℃. Phospholipase D catalyzes the substitution of the choline group in phosphatidylcholine with serine to generate phosphatidylserine, with simultaneous stirring during the conversion process.

[0040] S3. Separation was performed using column chromatography, and purification was achieved by adjusting the proportion of n-hexane as the mobile phase.

[0041] In step S1, microbial fermentation uses Streptomyces cinnamon, and in step S2, the catalytic reaction time is 6-8 hours. During the stirring process, the temperature of the mixture in different areas of the reactor 2 is monitored by the second type of temperature sensor 6 set on the functional component 43.

[0042] The foregoing has shown and described the basic principles, main features, and advantages of the present invention. It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the invention can be implemented in other specific forms without departing from its spirit or essential characteristics. Therefore, the embodiments should be considered illustrative and non-limiting in all respects, and the scope of the invention is defined by the appended claims rather than the foregoing description. Thus, all variations falling within the meaning and scope of equivalents of the claims are intended to be included within the present invention. No reference numerals in the claims should be construed as limiting the scope of the claims.

[0043] Furthermore, it should be understood that although this specification describes embodiments, not every embodiment contains only one independent technical solution. This narrative style is merely for clarity. Those skilled in the art should consider the specification as a whole, and the technical solutions in each embodiment can also be appropriately combined to form other embodiments that can be understood by those skilled in the art.

Claims

1. An apparatus for the catalytic preparation of phosphatidylserine, characterized in that: The reactor includes a fixed base (1), a reactor (2) fixedly mounted on the fixed base (1), a stirring device installed inside the reactor (2), and a heating device (5) installed inside the reactor (2). The reactor (2) is provided with a first inlet (21), a second inlet (22), and an outlet (23). An observation window (24) is provided on the peripheral side of the reactor (2). The observation window (24) extends into the inner cavity (25) of the reactor (2) and is covered by tempered glass. The inner cavity (25) of the reactor (2) is... 5) A first-class temperature sensor (26) is provided. The stirring device includes a central stirring device (3) and an edge stirring device (4). The central stirring device (3) is driven by a rotary motor (31) and simultaneously drives the edge stirring device (4) to mix and stir the materials. The central stirring device (3) is located at the central axis of the reactor (2), and the edge stirring device (4) is located at the inner side wall of the reactor (2). The central stirring device (3) and the edge stirring device (4) work together to stir. The central stirring device (3) includes a rotary motor (31), a rotating shaft (32) fixedly connected to the drive end of the rotary motor (31), and a stirring ring assembly (33) disposed on the rotating shaft (32). The stirring ring assembly (33) includes several central support rods (331) fixedly connected to the rotating shaft (32) and a central stirring ring (332) fitted onto each of the central support rods (331). The central support rods (331) are inclined to the horizontal plane, and the angle between them and the rotating shaft (32) is 100°-120°. The central stirring ring (332) is fixedly connected to the free end of each of the central support rods (331). A first top block (3321) is disposed on one side of the central stirring ring (332) near the inner cavity (25) of the reactor (2). The edge stirring device (4) includes an annular locking groove (41) opened on the inner side of the reactor (2), an annular functional plate (42) locked in the annular locking groove (41), and a functional ornament (43) disposed on the annular functional plate (42). The annular functional plate (42) is provided with an annular functional cavity (421), and the functional ornament (43) is disposed in the annular functional cavity (421). The annular functional plate (42) is surrounded on one side near the rotating shaft (32). There are several first connecting slots (423), each of which extends into the annular functional cavity (421). The upper and lower surfaces of the annular functional plate (42) are respectively provided with second connecting slots (422) and third connecting slots (424). The functional ornament (43) extends outward through the first connecting slots (423), the second connecting slots (422) and the third connecting slots (424). The functional ornament (43) is driven to work by the first top block (3321).

2. The apparatus for catalytic preparation of phosphatidylserine according to claim 1, characterized in that: The functional ornament (43) is provided with a rotating shaft (4322), which is rotatably disposed in the annular functional cavity (421) through the rotating shaft (4322). The functional ornament (43) includes an upper swing area (431), a central rotation area (432) and a lower swing area (433). The functional ornament (43) is provided with a torsion spring, which controls the movement stroke of the functional ornament (43) and resets it.

3. The apparatus for catalytic preparation of phosphatidylserine according to claim 2, characterized in that: The upper swing area (431) of the functional ornament (43) extends outward through the second connecting groove (422). A second top block (4321) is provided on the central rotating area (432) of the functional ornament (43). The second top block (4321) extends outward through the first connecting groove (423). The lower swing area (433) of the functional ornament (43) extends outward through the third connecting groove (424). The diameter of the opening of the second connecting groove (422) is greater than the width of the upper swing area (431), and the diameter of the opening of the third connecting groove (424) is greater than the width of the lower swing area (433).

4. The apparatus for catalytic preparation of phosphatidylserine according to claim 3, characterized in that: In the initial state, the central rotation area (432) is set perpendicular to the horizontal plane, the upper swing area (431) has an angle of 45°-60° with the horizontal plane, and the lower swing area (433) has an angle of 65°-85° with the horizontal plane. The functional ornament (43) is rotated and swung by the first top block (3321) pressing against the second top block (4321).

5. The apparatus for catalytic preparation of phosphatidylserine according to claim 4, characterized in that: When the first top block (3321) and the second top block (4321) come into contact with each other, the first top block (3321) pushes the second top block (4321) inward, and the rotatable functional ornament (43) rotates. The contact between the first top block (3321) and the second top block (4321) is intermittent, which drives the functional ornament (43) to rotate intermittently. The upper swing area (431) rotates away from the rotation axis (32), and the lower swing area (433) rotates closer to the rotation axis (32).

6. The apparatus for catalytic preparation of phosphatidylserine according to claim 5, characterized in that: A second type of temperature sensor (6) is provided at the free end of both the upper swing region (431) and the lower swing region (433).

7. A method for catalytically preparing phosphatidylserine, wherein the preparation is carried out using the apparatus described in claim 6, characterized in that: Includes the following steps: S1. Raw material preparation: Phospholipase D is prepared by microbial fermentation using soybean lecithin and L-serine as raw materials. S2. The apparatus for catalytic preparation of phosphatidylserine operates at pH 6.5-7.5 and 37℃. Phospholipase D catalyzes the substitution of the choline group in phosphatidylcholine with serine to generate phosphatidylserine, with simultaneous stirring during the conversion process. S3. Separation was performed using column chromatography, and purification was achieved by adjusting the proportion of n-hexane as the mobile phase.

8. The method for preparing phosphatidylserine by catalysis according to claim 7, characterized in that: In step S1, microbial fermentation uses Streptomyces cinnamon. In step S2, the catalytic reaction time is 6-8 hours. During the stirring process, the temperature of the mixture in different areas of the reactor (2) is monitored by the temperature sensor set on the functional component (43).

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