(R)-citronellal separation device and method

Abstract

The invention discloses a (R)-citronellal separation device and method. The (R)-citronellal separation device comprises a pretreatment mechanism, a first filtering mechanism, a coalescence separator, an adsorption mechanism, a second filtering mechanism, a rectifying tower, a light component removal tower and a heavy component removal tower which are sequentially connected. According to the invention, the problems of incomplete impurity removal, low product purity, complex product recovery process, difficulty in realizing continuous operation and inadaptability to industrial large-scale production requirements in the prior art can be solved, so that the purposes of efficiently removing impurities, improving the product purity and yield, and being mature in process and easy to industrialize are achieved. The method can be used for separating and purifying the (R)-citronellal, the purity of the (R)-citronellal product can reach 99.5% or above, and continuous operation can be achieved.

Description

Technical Field

[0001] This application relates to the fields of biosynthesis and separation technology, and in particular to (R)-citronellol separation apparatus and method. Background Technology

[0002] (R)-Citronellol is an important monoterpene aldehyde compound with a refreshing lemon aroma. It is not only a key component of high-end fragrances but also a core intermediate in the synthesis of menthol, ionone, and other pharmaceutical and daily chemical products. The market demand for high-purity (>99.5%) (R)-citronellol is increasingly urgent. The preparation methods for (R)-citronellol mainly include chemical synthesis and biocatalysis (enzymatic methods). Among these, biocatalysis has become a research hotspot in recent years due to its advantages such as mild reaction conditions, high stereoselectivity, and environmental friendliness. However, in the biocatalytic preparation process, the cell catalytic solution has a complex composition, containing a large number of bacterial cell fragments, nucleic acids, proteins, and polysaccharides, among other impurities. These impurities have similar physicochemical properties to (R)-citronellol, making subsequent separation extremely difficult.

[0003] Traditional separation and purification processes often employ single extraction, filtration, or distillation methods. These methods have the following problems: impurities are not completely removed, especially viscous impurities such as proteins and nucleic acids, which easily adhere to the product, making it difficult to achieve a product purity of over 99%; the product loss rate during purification is high, and the yield is usually below 80%; the process steps are scattered, making it difficult to achieve continuous operation and adapt to the needs of large-scale industrial production. Summary of the Invention

[0004] Therefore, it is necessary to provide a (R)-citronellol separation device that can efficiently remove impurities, improve product purity and yield, and is easy to industrialize.

[0005] One embodiment of this application provides an (R)-citronellol separation device.

[0006] A (R)-citronellol separation device includes a pretreatment unit, a first filtration unit, a coalescing separator, an adsorption unit, a second filtration unit, a distillation column, a light-weight removal column, and a heavy-weight removal column connected in sequence. The pretreatment unit is used for enzyme pretreatment by adding nuclease and protease to a cell catalytic solution. The first filtration unit is used for a first solid-liquid separation of the pretreated solution after enzyme pretreatment to obtain a pretreated filtrate. The coalescing separator is used for continuous phase separation of the pretreated filtrate to obtain an organic phase solution. The adsorption unit is used for decolorizing and removing impurities from the organic phase solution. The second filtration unit is used for a second solid-liquid separation of the decolorized and impurity-removed mixture to obtain a purified solution. The distillation column is used for distillation of the purified solution to obtain a hexane-removed solution. The light-weight removal column is used for light-weight removal treatment of the hexane-removed solution to obtain light-weight citronellol. The heavy-weight removal column is used for heavy-weight removal treatment of the light-weight citronellol to obtain (R)-citronellol.

[0007] In some embodiments, the (R)-citronellol separation device further includes a filtrate storage tank connected between the first filtration unit and the coalescing separator, the filtrate storage tank being used to store the pretreated filtrate.

[0008] In some embodiments, the filtrate storage tank is connected to a stirring mechanism for stirring the pretreated filtrate inside the filtrate storage tank.

[0009] In some embodiments, the (R)-citronellol separation device further includes an aqueous phase storage tank connected to the coalescing separator, the aqueous phase being used to store the aqueous phase after continuous phase separation treatment by the coalescing separator.

[0010] In some embodiments, the (R)-citronellol separation device further includes an organic phase storage tank connected between the coalescing separator and the second filtration mechanism, the organic phase storage tank being used to store the organic phase solution after continuous phase separation treatment by the coalescing separator.

[0011] In some embodiments, the (R)-citronellol separation device further includes a post-purification solution storage tank connected between the second filtration mechanism and the distillation column, the post-purification solution storage tank being used to store the post-purification solution after the second solid-liquid separation.

[0012] In some embodiments, the (R)-citronellol separation device further includes a product storage tank connected to the deweighting tower, the product storage tank being used to store the (R)-citronellol obtained from the deweighting treatment.

[0013] In some embodiments, the first filtration mechanism and the second filtration mechanism each independently include a plate and frame filter press.

[0014] In some embodiments, the distillation column has 6 to 10 theoretical plates, and the packing includes triangular spiral packing with a particle size of 3 mm to 5 mm.

[0015] In some embodiments, the light-weight removal tower has 20 to 30 theoretical trays and is filled with 3mm to 5mm triangular spiral packing.

[0016] In some embodiments, the theoretical number of trays in the deweight removal tower is 20 to 30, and the packing is 3mm to 5mm triangular spiral packing.

[0017] An embodiment of this application also provides a method for separating (R)-citronellol.

[0018] A method for separating (R)-citronellol includes the following steps:

[0019] Nuclease and protease were added to the cell catalytic solution for enzyme pretreatment to obtain a pretreated solution;

[0020] The pretreated liquid is subjected to a first solid-liquid separation to obtain a pretreated filtrate;

[0021] The pretreated filtrate was denatured and then subjected to continuous phase separation to obtain an organic phase solution.

[0022] The organic phase solution is subjected to decolorization and impurity removal treatment to obtain a purified mixture;

[0023] The impurity-removed mixture is subjected to a second solid-liquid separation to obtain a purified solution;

[0024] The purified solution is subjected to distillation to obtain a hexane-free solution;

[0025] The hexane-removed solution was subjected to a light-removal treatment to obtain light-removed citronellol;

[0026] Furthermore, the de-lightened citronellol is subjected to a de-heavy treatment to obtain (R)-citronellol.

[0027] In some embodiments, the amount of nuclease added is 0.01% to 0.5% of the mass of the cell catalytic solution.

[0028] In some embodiments, the amount of nuclease added is 0.05% to 0.1% of the mass of the cell catalytic solution.

[0029] In some embodiments, the amount of protease added is 0.1% to 2% of the mass of the cell catalytic solution.

[0030] In some embodiments, the amount of protease added is 0.2% to 0.5% of the mass of the cell catalytic solution.

[0031] In some embodiments, after adding the nuclease to the cell catalytic solution, the mixture is stirred at a temperature of 20°C to 40°C for at least 1 hour.

[0032] In some embodiments, after the protease is added to the cell catalytic solution, the mixture is stirred at a temperature of 20°C to 40°C for at least 1 hour.

[0033] In some embodiments, the denaturation treatment of the pretreated filtrate includes the following steps: heating the pretreated filtrate to 70°C~90°C for 1h~2h.

[0034] In some embodiments, continuous phase separation includes the following steps: controlling the residence time of the pretreated filtrate after denaturation treatment in the coalescence separator to be 0.2h to 1h.

[0035] In some embodiments, the filter element of the coalescing separator is controlled to have a filter element precision of 1μm to 5μm, made of polyester fiber, with a filter element structural strength > 0.7MPa, and the working pressure of the coalescing separator is controlled to be 0.3MPaA to 1MPaA.

[0036] In some embodiments, the decolorization and impurity removal treatment of the organic phase solution includes the following steps: adding activated carbon to the organic phase solution and heating to a temperature of 50°C~80°C, stirring for 1h~3h; wherein the activated carbon has a specific surface area of ​​1000m². 2 / g~1700m 2 / g, methylene blue adsorption value >180.

[0037] In some embodiments, the mass ratio of the organic phase solution to the activated carbon is (20~50):1.

[0038] In some embodiments, the first solid-liquid separation and the second solid-liquid separation are performed independently using a plate and frame filter press.

[0039] In some embodiments, the distillation process uses a distillation column, and the number of theoretical plates in the distillation column is controlled to be 6 to 10. The packing includes triangular spiral packing with a particle size of 3 mm to 5 mm. The top pressure of the column is 100 kPaA to 120 kPaA, and the bottom temperature is 95°C to 105°C.

[0040] In some embodiments, the light-light removal process uses a light-light removal tower, and the theoretical number of trays in the light-light removal tower is controlled to be 20 to 30, the packing is 3mm to 5mm triangular spiral packing, the top pressure is 1.5kPaA to 2kPaA, the bottom temperature is 120℃ to 130℃, and the reflux ratio is (10 to 15):1.

[0041] In some embodiments, the deweighting process uses a deweighting tower, and the theoretical number of trays in the deweighting tower is controlled to be 20 to 30, the packing is 3mm to 5mm triangular spiral packing, the top pressure is 1.5kPaA to 2kPaA, the bottom temperature is 120℃ to 130℃, and the reflux ratio is 1:(15 to 20).

[0042] In some embodiments, the (R)-citronellol separation method employs the (R)-citronellol separation device described in any of the above embodiments.

[0043] The aforementioned (R)-citronellol separation device solves the problems of incomplete impurity removal, low product purity, complex product recovery processes, and difficulty in achieving continuous operation, which are unsuitable for large-scale industrial production. It achieves efficient impurity removal, improved product purity and yield, and is a mature process easily scalable for industrial application. The (R)-citronellol separation device of this application can be used to separate and purify (R)-citronellol, producing a high-purity product that can be operated continuously.

[0044] The method for separating (R)-citronellol in this application includes: adding nuclease and protease to a cell catalytic solution containing (R)-citronellol for enzyme pretreatment; heating and denaturing the enzyme-pretreated (R)-citronellol cell catalytic solution, followed by separation of bacterial residue and other insoluble substances; thoroughly mixing the filtrate and performing continuous phase separation; adding activated carbon to the obtained organic phase solution, heating and stirring thoroughly for decolorization and impurity removal, and separating and removing the waste activated carbon; after impurity removal, the organic phase solution is subjected to continuous distillation to recover the solvent, separating light and heavy components of impurities, and finally obtaining (R)-citronellol with high purity. This method for separating (R)-citronellol can efficiently remove impurities from the (R)-citronellol cell catalytic solution, effectively improving product purity and solving the problems of high separation difficulty and low purification yield in traditional enzymatic preparation of citronellol. The process steps are simple, the technology is mature, and it is easy to industrialize. Attached Figure Description

[0045] To more clearly illustrate the technical solutions in the embodiments of this application, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0046] To gain a more complete understanding of this application and its beneficial effects, the following description will be provided in conjunction with the accompanying drawings. In the following description, the same reference numerals denote the same parts.

[0047] Figure 1 This is a schematic diagram of the (R)-citronellol separation device according to an embodiment of this application;

[0048] Figure 2 This is a schematic diagram of the (R)-citronellol separation method according to an embodiment of this application.

[0049] Explanation of reference numerals in the attached figures

[0050] 10. (R)-Citronellol Separation Unit; 100. Pretreatment Unit; 200. First Filtration Unit; 300. Coalescing Separator; 400. Adsorption Unit; 500. Second Filtration Unit; 600. Distillation Column; 700. Light Phase Removal Column; 800. Heavy Phase Removal Column; 901. Filtrate Storage Tank; 902. Aqueous Phase Storage Tank; 903. Organic Phase Storage Tank; 904. Impurity-Removed Solution Storage Tank; 905. Product Storage Tank; 1000. Stirring Unit. Detailed Implementation

[0051] To make the above-mentioned objectives, features, and advantages of this application more apparent and understandable, the specific embodiments of this application are described in detail below with reference to the accompanying drawings. Many specific details are set forth in the following description to provide a thorough understanding of this application. However, this application can be implemented in many other ways different from those described herein, and those skilled in the art can make similar modifications without departing from the spirit of this application. Therefore, this application is not limited to the specific embodiments disclosed below.

[0052] In the description of this application, it should be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", "axial", "radial", "circumferential", etc., indicating the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings, are only for the convenience of describing this application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation of this application.

[0053] In this application, unless otherwise expressly specified and limited, the terms "installation," "connection," "joining," and "fixing," etc., should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral part; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; they can refer to the internal communication of two components or the interaction between two components, unless otherwise expressly limited. Those skilled in the art can understand the specific meaning of the above terms in this application according to the specific circumstances.

[0054] In this application, unless otherwise expressly specified and limited, "above" or "below" the second feature can mean that the first feature is in direct contact with the second feature, or that the first feature is in indirect contact with the second feature through an intermediate medium. Furthermore, "above," "on top of," and "over" the second feature can mean that the first feature is directly above or diagonally above the second feature, or simply that the first feature is at a higher horizontal level than the second feature. "Below," "below," and "under" the second feature can mean that the first feature is directly below or diagonally below the second feature, or simply that the first feature is at a lower horizontal level than the second feature.

[0055] In the description of this application, "several" means one or more, "multiple" means two or more, "greater than," "less than," and "exceeding" are understood to exclude the stated number, while "above," "below," and "within" are understood to include the stated number. The use of "first" and "second" in the description is merely for distinguishing technical features and should not be construed as indicating or implying relative importance, or implicitly indicating the number of indicated technical features, or implicitly indicating the order of the indicated technical features.

[0056] In this document, "optionally," "optionally," and "optional" mean that something is optional, that is, it is selected from either "with" or "without." If multiple "options" appear in a technical solution, unless otherwise specified and there are no contradictions or mutual constraints, each "option" is independent. In this application, descriptions such as "optionally contains" and "optionally includes" indicate "contains or does not contain."

[0057] In this application, unless otherwise stated, the sum of the parts of each component in the composition may be 100 parts by weight. Unless otherwise specified, the percentages (including weight percentages) in this application are based on the total weight of the composition, and "wt%" in this document means mass percentage.

[0058] In this document, unless otherwise stated, the reaction steps may be performed in the order described herein or not. For example, other steps may be included between reaction steps, and the order of reaction steps may be appropriately interchanged. This is something that those skilled in the art can determine based on conventional knowledge and experience. Preferably, the reaction methods described herein are performed sequentially.

[0059] In this application, when numerical intervals (i.e., numerical ranges) are mentioned, unless otherwise specified, the distribution of selectable numerical values ​​within the numerical interval is considered continuous, and includes the two endpoints of the numerical interval (i.e., the minimum and maximum values), as well as every numerical value between these two endpoints. Unless otherwise specified, when a numerical interval refers only to integers within that numerical interval, it includes the two endpoint integers of the numerical range, as well as every integer between the two endpoints, which is equivalent to directly listing every integer. When multiple numerical ranges are provided to describe features or characteristics, these numerical ranges can be merged. In other words, unless otherwise specified, the numerical ranges disclosed in this application should be understood to include any and all subranges included therein. The "numerical value" in the numerical interval can be any quantitative value, such as a number, percentage, ratio, etc. The term "numerical interval" can be broadly included to include percentage intervals, ratio intervals, proportion intervals, etc.

[0060] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0061] This application provides an embodiment of a (R)-citronellol separation device 10 to solve at least one of the following technical problems in the preparation of (R)-citronellol using biocatalysis (enzymatic method) in the traditional technology: (1) The cell catalytic solution has a complex composition, containing a large number of bacterial cell fragments, nucleic acids, proteins and polysaccharides, etc. These impurities are similar to the physicochemical properties of (R)-citronellol, making subsequent separation extremely difficult. (2) When using a single extraction, filtration or distillation method for separation, the impurities are not completely removed, especially viscous impurities such as proteins and nucleic acids, which are easy to adhere to the product, making it difficult for the product purity to exceed 99%. (3) The product loss rate is high during the purification process, and the yield is usually less than 80%. (4) The process steps are scattered, making it difficult to achieve continuous operation and adapt to the needs of large-scale industrial production. The (R)-citronellol separation device 10 will be described below with reference to the accompanying drawings.

[0062] The (R)-citronellol separation device 10 provided in one embodiment of this application is exemplary; please refer to [link to example]. Figure 1As shown, Figure 1 This is a schematic diagram of the (R)-citronellol separation device 10 provided in one embodiment of this application. The (R)-citronellol separation device 10 of this application can be used to separate and purify (R)-citronellol, and the purity of the (R)-citronellol product can reach more than 99.5%. The process steps are simple and can be operated continuously.

[0063] To more clearly illustrate the structure of the (R)-citronellol separation device 10, the (R)-citronellol separation device 10 will be described below in conjunction with the accompanying drawings.

[0064] For example, please refer to Figure 1 As shown, an (R)-citronellol separation device 10 includes a pretreatment unit 100, a first filtration unit 200, a coalescing separator 300, an adsorption unit 400, a second filtration unit 500, a distillation column 600, a light component removal column 700, and a heavy component removal column 800 connected in sequence.

[0065] The system includes the following components: a pretreatment unit 100 for enzyme pretreatment by adding nucleases and proteases to the cell catalytic solution; a first filtration unit 200 for performing a first solid-liquid separation on the pretreated solution to obtain a pretreated filtrate, bacterial residue, and insoluble matter; a coalescence separator 300 for continuous phase separation of the pretreated filtrate to obtain an organic phase solution; an adsorption unit 400 for decolorizing and removing impurities from the organic phase solution; a second filtration unit 500 for performing a second solid-liquid separation on the decolorized and impurity-removed mixture to obtain a purified solution; a distillation column 600 for distilling the purified solution to obtain a hexane-removed solution; a light-weight removal column 700 for removing light-weight components from the hexane-removed solution to obtain light-weight citronellal; and a heavy-weight removal column 800 for removing heavy-weight components from the light-weight citronellal to obtain (R)-citronellal.

[0066] The aforementioned (R)-citronellol separation device 10 solves the problems of incomplete impurity removal, low product purity, complex product recovery processes, and difficulty in achieving continuous operation, which are unsuitable for large-scale industrial production. It achieves efficient impurity removal, improved product purity and yield, and is a mature process easily industrialized. The (R)-citronellol separation device 10 of this application can be used to separate and purify (R)-citronellol, and the purity of the (R)-citronellol product can reach over 99.5%, enabling continuous operation.

[0067] In some embodiments, an adsorption mechanism 1000 is connected to the adsorption mechanism 400.

[0068] In some of these implementations, please refer to Figure 1As shown, the (R)-citronellol separation device 10 also includes a filtrate storage tank 901. The filtrate storage tank 901 is connected between the first filtration mechanism 200 and the coalescing separator 300, and is used to store the pretreated filtrate.

[0069] In some of these implementations, please refer to Figure 1 As shown, the filtrate storage tank 901 is connected to a stirring mechanism 1000 for stirring the pretreated filtrate in the filtrate storage tank 901.

[0070] In some of these implementations, please refer to Figure 1 As shown, the (R)-citronellol separation device 10 also includes an aqueous phase storage tank 902. The aqueous phase storage tank 902 is connected to the coalescing separator 300 and is used to store the aqueous phase after continuous phase separation treatment by the coalescing separator 300.

[0071] In some of these implementations, please refer to Figure 1 As shown, the (R)-citronellol separation device 10 also includes an organic phase storage tank 903. The organic phase storage tank 903 is connected between the coalescing separator 300 and the second filtration unit 500. The organic phase storage tank 903 is used to store the organic phase solution after continuous phase separation treatment by the coalescing separator 300.

[0072] It should be noted that when the organic phase storage tank 903 is provided, the adsorption mechanism 400 can be omitted, that is, the organic phase storage tank 903 can be used as the adsorption mechanism 400. In this case, after omitting the adsorption mechanism 400, the corresponding stirring mechanism 1000 is provided on the organic phase storage tank 903.

[0073] In some of these implementations, please refer to Figure 1 As shown, the (R)-citronellol separation device 10 also includes a post-purification solution storage tank 904. The post-purification solution storage tank 904 is connected between the second filtration mechanism 500 and the distillation column 600, and is used to store the post-purification solution after the second solid-liquid separation.

[0074] In some of these implementations, please refer to Figure 1 As shown, the (R)-citronellol separation device 10 also includes a product storage tank 905. The product storage tank 905 is connected to the deweighting tower 800 and is used to store the (R)-citronellol obtained from the deweighting treatment.

[0075] In some embodiments, the first filtration unit 200 and the second filtration unit 500 each independently include a plate and frame filter press.

[0076] In some embodiments, the distillation column 600 has 6 to 10 theoretical plates, and the packing includes triangular spiral packing with a particle size of 3 mm to 5 mm.

[0077] In some embodiments, the light-removal tower 700 has 20 to 30 theoretical trays and is filled with 3mm to 5mm triangular spiral packing.

[0078] In some embodiments, the theoretical number of trays in the deweight removal tower 800 is 20 to 30, and the packing is 3mm to 5mm triangular spiral packing.

[0079] In some embodiments, the pretreatment unit 100 includes a reaction vessel, the volume of which can be set as needed, such as 30L, 50L, etc.

[0080] An embodiment of this application also provides a method for separating (R)-citronellol.

[0081] Please see Figure 2 As shown, Figure 2 This is a schematic flowchart of a (R)-citronellol separation method according to an embodiment of this application. The (R)-citronellol separation method includes the following steps:

[0082] S10. Nuclease and protease are added to the cell catalytic solution for enzyme pretreatment to obtain the pretreated solution.

[0083] S20. Perform a first solid-liquid separation on the pretreated liquid to obtain pretreated filtrate, bacterial residue, and insoluble matter.

[0084] S30. After denaturation treatment of the pretreated filtrate, continuous phase separation treatment is performed to obtain an organic phase solution.

[0085] S40. The organic phase solution is decolorized and impurity removed to obtain a purified mixture.

[0086] S50. Perform a second solid-liquid separation on the impurity-removed mixture to obtain the impurity-removed solution.

[0087] S60. The purified solution is subjected to distillation to obtain a hexane-free solution.

[0088] S70. The hexane-removed solution is subjected to a light-removing treatment to obtain light-removed citronellal.

[0089] S80. (R)-citronellol is obtained by removing the heavy weight of citronellol.

[0090] In some embodiments, the cell catalytic solution comprises 2.5wt%~3wt% wet bacterial cells, 6wt%~12wt% n-hexane, 8wt%~10wt% sodium gluconate, 70wt%~75wt% pure water, 5wt%~8wt% citronellal, and 0.06wt%~0.1wt% impurities.

[0091] In some embodiments, during S10, when adding nuclease and protease, nuclease can be added first, followed by protease.

[0092] In some embodiments, the amount of nuclease added is 0.01% to 0.5% of the mass of the cell catalytic solution. For example, the amount of nuclease added as a percentage of the mass of the cell catalytic solution includes, but is not limited to: 0.01%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, or any range between the foregoing.

[0093] In some embodiments, the amount of nuclease added is 0.05% to 0.1% of the mass of the cell catalytic solution.

[0094] In some embodiments, the amount of protease added is 0.1% to 2% of the mass of the cell catalytic solution. For example, the amount of protease added as a percentage of the mass of the cell catalytic solution includes, but is not limited to: 0.1%, 0.15%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, or any range between the foregoing.

[0095] In some embodiments, the amount of protease added is 0.2% to 0.5% of the mass of the cell catalytic solution.

[0096] In some embodiments, after adding nuclease to the cell catalytic solution, the mixture is stirred at 20°C to 40°C and at a speed of 300 rpm to 500 rpm for at least 1 hour.

[0097] In some embodiments, after adding the protease to the cell catalytic solution, the mixture is stirred at a temperature of 20°C to 40°C and a speed of 300 rpm to 500 rpm for at least 1 hour.

[0098] In some embodiments, the denaturation treatment of the pretreated filtrate includes the following steps: heating the pretreated filtrate to 70°C~90°C for 1h~2h. For example, the heating temperature of the pretreated filtrate during denaturation treatment includes, but is not limited to, 70°C, 75°C, 80°C, 85°C, 90°C, or any range between the foregoing. For example, the denaturation time of the pretreated filtrate during denaturation treatment includes, but is not limited to, 1h, 1.2h, 1.5h, 1.8h, 2h, or any range between the foregoing.

[0099] In some embodiments, continuous phase separation includes the following steps: controlling the residence time of the pretreated filtrate after denaturation treatment in the coalescing separator 300 to be 0.2h to 1h, preferably 0.3h to 0.5h. For example, the value of the residence time of the pretreated filtrate after denaturation treatment in the coalescing separator 300 includes, but is not limited to: 0.2h, 0.3h, 0.5h, 0.8h, 1h, or any range between the foregoing.

[0100] In some embodiments, the filter element precision of the coalescing separator 300 is controlled to be 1μm to 5μm, made of polyester fiber, with a filter element structural strength > 0.7MPa, and the operating pressure of the coalescing separator 300 is controlled to be 0.3MPaA to 1MPaA. For example, the value of the filter element precision of the coalescing separator 300 includes, but is not limited to: 1μm, 2μm, 3μm, 4μm, 5μm, or any range between the aforementioned. Similarly, the value of the operating pressure of the coalescing separator 300 includes, but is not limited to: 0.3MPaA, 0.4MPaA, 0.5MPaA, 0.6MPaA, 0.7MPaA, 0.8MPaA, 0.9MPaA, 1MPaA, or any range between the aforementioned.

[0101] In some embodiments, the decolorization and impurity removal treatment of the organic phase solution includes the following steps: adding activated carbon to the organic phase solution and heating to a temperature of 50°C~80°C, stirring for 1h~3h; wherein the specific surface area of ​​the activated carbon is 1000m². 2 / g~1700m 2 / g, methylene blue adsorption value >180. For example, when decolorizing and removing impurities from an organic phase solution, the temperature at which activated carbon is added and heated is included, but not limited to: 50℃, 55℃, 60℃, 65℃, 70℃, 75℃, 80℃, or any range between the foregoing. For example, when decolorizing and removing impurities from an organic phase solution, the stirring time after adding activated carbon is included, but not limited to: 1h, 1.2h, 1.5h, 1.8h, 2h, or any range between the foregoing.

[0102] In some embodiments, the decolorization and impurity removal treatment of the organic phase solution includes the following steps: adding activated carbon to the organic phase solution and heating it to a temperature of 60°C~70°C, and stirring for 1.5h~2h.

[0103] In some embodiments, the mass ratio between the organic phase solution and activated carbon is (20~50):1. For example, the mass ratio between the organic phase solution and activated carbon includes, but is not limited to: 20:1, 25:1, 30:1, 35:1, 40:1, 45:1, 50:1 or any range between the foregoing.

[0104] In some embodiments, the first solid-liquid separation and the second solid-liquid separation are performed independently using a plate and frame filter press.

[0105] In some embodiments, during distillation, a distillation column 600 is used, and the number of theoretical plates in the distillation column 600 is controlled to be 6 to 10. The feed plate is located at position 3, and the packing includes triangular spiral packing with a particle size of 3 mm to 5 mm. The top pressure is 100 kPaA to 120 kPaA, and the bottom temperature is 95°C to 105°C. The bottom temperature is preferably 100 ± 2°C.

[0106] In some embodiments, during the removal of light components, a light component removal tower 700 is used, and the theoretical number of trays in the light component removal tower 700 is controlled to be 20 to 30, the feed tray position is 15, the packing is 3mm to 5mm triangular spiral packing, the top pressure is 1.5kPaA to 2kPaA, the bottom temperature is 120℃ to 130℃, and the reflux ratio is (10 to 15):1.

[0107] In some embodiments, during the deweighting process, a deweighting tower 800 is used, and the theoretical number of trays of the deweighting tower 800 is controlled to be 20 to 30, the feed tray position is 10, the packing is 3mm to 5mm triangular spiral packing, the top pressure is 1.5kPaA to 2kPaA, the bottom temperature is 120℃ to 130℃, and the reflux ratio is 1:(15 to 20).

[0108] In some of these embodiments, the (R)-citronellol separation method employs the (R)-citronellol separation device 10 described in any of the above embodiments.

[0109] Example 1

[0110] This embodiment provides a method for separating (R)-citronellol.

[0111] The relevant equipment and raw material parameters in this embodiment are as follows:

[0112] R-citronellol cell catalytic solution was prepared according to the enzyme-catalyzed preparation method of (R)-citronellol in Example 4 of patent CN118272331B. Specific examples are as follows:

[0113] 100 mL reaction system: 500 mM (E / Z)-citral ((E / Z)-citral was added as a 3 M substrate solution prepared with n-hexane), 510 mM glucose (added as an aqueous glucose solution), 2 g / L glucose dehydrogenase, 0.2 mM NADH (added as an aqueous NADH solution), n-hexane was added to make the volume percentage of n-hexane in the system 20%, 7.5 g of frozen stem cells of mutant CvDH-A181R obtained in Example 3 (equivalent to 0.99 g / g of (E / Z)-citral), 100 mM phosphate buffer (pH 8.5) was added to make up to 100 mL, and the reaction was carried out at 35 °C and stirred at 200 rpm for 4 hours.

[0114] The composition of R-citronellol cell catalytic solution is shown in Table 1 below:

[0115] Table 1

[0116]

[0117] Activated carbon: specific surface area of ​​1000 m² 2 / g~1700m 2 / g, methylene blue adsorption value >180, Yantai Tongyi Environmental Protection Technology Co., Ltd.

[0118] Coalescing separator 300: Filter element size is 1μm~5μm, made of polyester fiber; filter element structural strength is >0.7MPa, and the working pressure of coalescing separator 300 is 0.3MPaA~1MPaA.

[0119] Protease: Brand name: BLAZE EVITY 100T, Product name: Yongwei Protease 100T, Nominal activity >100KNPU-B / g, Manufacturer: Novozymes (China) Biotechnology Co., Ltd.

[0120] Nuclease: Brand name Everis TM Next, liquid, manufactured by Novozymes (China) Biotechnology Co., Ltd.

[0121] The (R)-citronellol separation method in this embodiment adopts... Figure 1 The (R)-citronellol separation device 10 shown includes the following steps:

[0122] S100. Take 15 kg of cell catalytic solution containing (R)-citronellol (C: the mass concentration of (R)-citronellol in the cell catalytic solution is 6%) and add it to a 30 L pretreatment unit 100 (reactor). Turn on the stirring mechanism 1000 connected to the pretreatment unit 100, and control the speed at 300 rpm to ensure sufficient mass transfer. Control the temperature of the cell catalytic solution at 22℃. After the temperature stabilizes, add 7.5 g of nuclease to the cell catalytic solution and stir at 22℃ for 1.5 h. Add 30 g of protease to the cell catalytic solution and stir at 22℃ for 1.5 h to obtain the pretreatment solution.

[0123] S200. The pretreated filtrate is heated to 70℃ and denatured for 2 hours. A plate and frame filter press is used to perform the first solid-liquid separation on the denatured pretreated filtrate to obtain the pretreated filtrate, bacterial residue, and insoluble matter. The bacterial residue and insoluble matter are discharged. The pretreated filtrate weighs approximately 14256g. The composition of the pretreated filtrate is shown in Table 2.

[0124] Table 2

[0125]

[0126] S300: The filter element of the coalescing separator 300 is set to a filter accuracy of 4μm, made of polyester fiber, with a filter element structural strength > 0.7MPa. The operating pressure of the coalescing separator 300 is controlled at 0.5MPaA. The residence time of the pretreated filtrate after denaturation treatment in the coalescing separator 300 is controlled at 0.3h, and continuous phase separation is performed to obtain an organic phase solution and an aqueous phase. The aqueous phase enters the aqueous phase storage tank 902 for storage.

[0127] The aqueous phase enters the aqueous phase storage tank 902, and the composition of the aqueous phase is shown in Table 3-1:

[0128] Table 3-1

[0129]

[0130] The organic phase solution enters the organic phase storage tank 903. The mass of the organic phase solution in the organic phase storage tank 903 is approximately 2165g. The composition of the organic phase solution is shown in Table 3-2.

[0131] Table 3-2

[0132]

[0133] S400. Add 65g of activated carbon to the organic phase storage tank 903, start stirring at 300rpm, heat to 60℃, and continue stirring for 1.5h to decolorize and remove impurities from the organic phase solution.

[0134] S500 uses a plate and frame filter press to perform a second solid-liquid separation on the decolorized and impurity-removed mixture (also known as the activated carbon-containing organic phase) to obtain a post-impurity-removed solution and waste activated carbon. Approximately 120g of waste activated carbon is discharged for treatment. Approximately 2000g of post-impurity-removed solution enters the post-impurity-removed solution storage tank 904, and the remaining post-impurity-removed solution remains in the equipment.

[0135] The composition of the solution after impurity removal is shown in Table 4:

[0136] Table 4

[0137]

[0138] The composition of the waste activated carbon is shown in Table 5:

[0139] Table 5

[0140]

[0141] Repeat steps S100 to S500 three times to obtain approximately 6 kg of purified solution. Use approximately 1 kg of purified solution to establish equilibrium in the continuous distillation system, and use 5 kg of purified solution for the entire process material balance calculation.

[0142] The S600 distillation column, after impurity removal, performs distillation on the purified solution to obtain a dehexane solution and n-hexane, with the n-hexane being discharged externally. The distillation column 600 has 6 theoretical trays, with the feed tray at position 3. The packing consists of triangular spiral packing with a particle size of 4 mm. The top pressure is 100 kPaA, and the bottom temperature is 100℃. At the top of the distillation column 600, 3226.69 g of n-hexane is obtained, and the n-hexane content in the bottoms is <50 mg / kg. At the bottoms, 1952.84 g of the dehexane solution is obtained.

[0143] S700 and the light component removal tower 700 process the hexane removal solution to obtain light-removed citronellal and light component impurities, which are discharged externally. The light component removal tower 700 has 20 theoretical trays, feed tray position 15, 3mm triangular spiral packing, a top pressure of 1.5 kPaA, a bottom temperature of 125℃, and a reflux ratio of 10:1. The top of the light component removal tower 700 yields 102.82 g of R-citronellal and light component impurities, while the bottom yields 1850 g of light-removed citronellal.

[0144] The S800 and 800 heavy component removal towers process citronellol to obtain (R)-citronellol and heavy components, with the heavy component impurities being discharged externally. The S800 heavy component removal tower has a theoretical number of 20 trays, a feed tray position of 10, and uses 3mm triangular spiral packing. The top pressure is 1.5 kPaA, the bottom temperature is 125℃, and the reflux ratio is 1:15. The top of the S800 heavy component removal tower yields 1720g of R-citronellol product with a purity of 99.85%, while the bottom yields 128g of citronellol and heavy component impurities.

[0145] The purity of (R)-citronellol prepared in this embodiment was tested by gas chromatography: HP-5 column (30m×0.32mm×0.25μm), split ratio 10:1, injection temperature: 225℃; FID detector temperature: 250℃; carrier gas: nitrogen, 50mL / min; air flow rate: 300mL / min; hydrogen flow rate: 20mL / min; column flow rate: 0.5mL / min; injection volume: 1μL; initial column temperature: 100℃, maintained for 2min, then increased to 150℃ at a rate of 10℃ / min and maintained for 5min. The purity of (R)-citronellol was calculated using the area method.

[0146] The yield of (R)-citronellol in the entire process is calculated by multiplying the yield (Y1) of (R)-citronellol in steps S100-S500 and the yield (Y2) of (R)-citronellol in steps S600-S800. 总 ).

[0147] The formula for calculating the purification yield of (R)-citronellol is as follows:

[0148] Y 总 =Y1×Y2;

[0149] Y1= ;

[0150] Y2= ;

[0151] Wherein, m1: weight of the solution after impurity removal in steps S100-S500; m2: weight of (R)-citronellol solution in steps S500-S800; m3: weight of the solution after impurity removal in steps S500-S800; w1: purity of (R)-citronellol in the solution after impurity removal; w2: purity of (R)-citronellol in the (R)-citronellol solution; M: volume of cell catalytic liquid; C: mass concentration of (R)-citronellol in cell catalytic liquid.

[0152] Calculation in this embodiment

[0153] Y1=(2071.81×37.25%) / (15000×6%)×100%=85.75%;

[0154] Y2=(1720×99.85) / (5000×37.25%)=92.21%;

[0155] (R)-Citronellol purification yield (%) Y 总 =Y1×Y2=79%.

[0156] Comparative Example 1

[0157] Comparative Example 1 provides a method for separating and purifying (R)-citronellol from R-citronellol cell catalytic fluid.

[0158] The composition of the cell catalytic solution in the reactor is shown in Table 6:

[0159] Table 6

[0160]

[0161] The temperature of the cell catalytic solution was raised to 70°C and maintained for 2 hours. After the heat treatment, the solution in the reactor was transferred to a small plate and frame filter press for filtration. During the separation process, it was found that the catalytic solution had a high viscosity and was difficult to separate.

[0162] The (R)-citronellol separation method of this application includes: sequentially adding nuclease and protease to a cell catalytic solution containing (R)-citronellol for enzyme pretreatment; heating and denaturing the enzyme-pretreated (R)-citronellol cell catalytic solution, followed by separation of bacterial residue and other insoluble substances; performing continuous phase separation after thorough mixing of the filtrate; adding activated carbon to the obtained organic phase solution, heating and stirring thoroughly for decolorization and impurity removal, and removing the waste activated carbon; and continuously distilling the organic phase solution after impurity removal to recover the solvent, separating light and heavy components of impurities, and finally obtaining (R)-citronellol with a purity >99.5%. This (R)-citronellol separation method can efficiently remove impurities from the R-citronellol cell catalytic solution, effectively improving product purity and solving the problems of high separation difficulty and low purification yield in traditional enzymatic preparation of citronellol. The process steps of this application are simple, the process is mature, and it is easy to industrialize.

[0163] In the above embodiments, the descriptions of each embodiment have different focuses. For parts not described in detail in a certain embodiment, please refer to the relevant descriptions in other embodiments.

[0164] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0165] The embodiments described above are merely illustrative of several implementation methods of this application, and while the descriptions are specific and detailed, they should not be construed as limiting the scope of this patent application. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this application, and these all fall within the protection scope of this application. Therefore, the protection scope of this patent application should be determined by the appended claims.

Claims

1. A (R)-citronellol separation device, characterized in that, The system comprises, in sequence, a pretreatment unit, a first filtration unit, a coalescing separator, an adsorption unit, a second filtration unit, a distillation column, a light-weight removal column, and a heavy-weight removal column. The pretreatment unit is used to obtain a pretreatment solution by adding nuclease and protease to a cell catalytic solution for enzyme pretreatment. The first filtration unit is used to perform a first solid-liquid separation on the pretreatment solution to obtain a pretreatment filtrate. The coalescing separator is used to perform continuous phase separation on the pretreatment filtrate to obtain an organic phase solution. The adsorption unit is used to decolorize and remove impurities from the organic phase solution to obtain a purified mixture. The second filtration unit is used to perform a second solid-liquid separation on the purified mixture to obtain a purified solution. The distillation column is used to distill the purified solution to obtain a hexane-removed solution. The light-weight removal column is used to remove light-weight components from the hexane-removed solution to obtain light-weight citronellal. The heavy-weight removal column is used to remove heavy components from the light-weight citronellal to obtain (R)-citronellal.

2. The (R)-citronellol separation device according to claim 1, characterized in that, It also meets at least one of the following conditions: (1) It also includes a filtrate storage tank, which is connected between the first filtration mechanism and the coalescing separator, and the filtrate storage tank is used to store the pretreated filtrate; Optionally, the filtrate storage tank is connected to a stirring mechanism for stirring the pretreated filtrate in the filtrate storage tank; (2) It also includes an aqueous phase storage tank, which is connected to the coalescing separator and is used to store the aqueous phase after continuous phase separation treatment by the coalescing separator; (3) It also includes an organic phase storage tank, which is connected between the coalescing separator and the second filtration mechanism. The organic phase storage tank is used to store the organic phase solution after continuous phase separation treatment by the coalescing separator. (4) It also includes a storage tank for the purified solution, which is connected between the second filtration mechanism and the distillation column, and is used to store the purified solution after the second solid-liquid separation; (5) It also includes a product storage tank, which is connected to the deweighting tower and is used to store the (R)-citronellol obtained by the deweighting treatment.

3. The (R)-citronellol separation device according to claim 1 or 2, characterized in that, It also meets at least one of the following conditions: (1) The first filtration mechanism and the second filtration mechanism each independently include a plate and frame filter press; (2) The theoretical number of plates in the distillation column is 6 to 10, and the packing includes triangular spiral packing with a particle size of 3 mm to 5 mm; (3) The theoretical number of trays in the light-light removal tower is 20 to 30, and the packing is 3mm to 5mm triangular spiral packing; (4) The theoretical number of trays in the deweight removal tower is 20 to 30, and the packing is 3mm to 5mm triangular spiral packing.

4. A method for separating (R)-citronellol, characterized in that, Includes the following steps: Nuclease and protease were added to the cell catalytic solution for enzyme pretreatment to obtain a pretreated solution; The pretreated liquid is subjected to a first solid-liquid separation to obtain a pretreated filtrate; The pretreated filtrate was denatured and then subjected to continuous phase separation to obtain an organic phase solution. The organic phase solution is subjected to decolorization and impurity removal treatment to obtain a purified mixture; The impurity-removed mixture is subjected to a second solid-liquid separation to obtain a purified solution; The purified solution is subjected to distillation to obtain a hexane-free solution; The hexane-removed solution was subjected to a light-removal treatment to obtain light-removed citronellol; Furthermore, the de-lightened citronellol is subjected to a de-heavy treatment to obtain (R)-citronellol.

5. The (R)-citronellol separation method according to claim 4, characterized in that, It also satisfies at least one of the following conditions: (1) The amount of nuclease added is 0.01% to 0.5% of the mass of the cell catalytic solution; Optionally, the amount of nuclease added is 0.05% to 0.1% of the mass of the cell catalytic solution; (2) The amount of the protease added is 0.1% to 2% of the mass of the cell catalytic solution; Optionally, the amount of the protease added is 0.2% to 0.5% of the mass of the cell catalytic solution.

6. The (R)-citronellol separation method according to claim 4, characterized in that, It also satisfies at least one of the following conditions: (1) After adding the nuclease to the cell catalytic solution, stir at 20℃~40℃ for at least 1 hour; (2) After adding the protease to the cell catalytic solution, stir at 20°C to 40°C for at least 1 hour.

7. The (R)-citronellol separation method according to any one of claims 4 to 6, characterized in that, It also satisfies at least one of the following conditions: (1) When denaturing the pretreated filtrate, the following steps are included: heating the pretreated filtrate to 70℃~90℃ and denaturing it for 1h~2h. (2) When performing continuous phase separation, the following steps are included: controlling the residence time of the pretreated filtrate after denaturation treatment in the coalescence separator to be 0.2h~1h; Optionally, the filter element precision of the coalescing separator is controlled to be 1μm~5μm, the filter element structural strength is >0.7MPa, and the working pressure of the coalescing separator is controlled to be 0.3MPaA~1MPaA.

8. The (R)-citronellol separation method according to any one of claims 4 to 6, characterized in that, It also satisfies at least one of the following conditions: (1) The decolorization and impurity removal treatment of the organic phase solution includes the following steps: adding activated carbon to the organic phase solution and heating it to a temperature of 50℃~80℃, stirring for 1h~3h; wherein the specific surface area of ​​the activated carbon is 1000m². 2 / g~1700m 2 / g, methylene blue adsorption value >180; Optionally, the mass ratio of the organic phase solution to the activated carbon is (20~50):1; (2) When performing the first solid-liquid separation and the second solid-liquid separation, a plate and frame filter press is used independently.

9. The (R)-citronellol separation method according to any one of claims 4 to 6, characterized in that, It also satisfies at least one of the following conditions: (1) During the distillation process, a distillation column is used, and the theoretical number of plates of the distillation column is controlled to be 6 to 10. The packing includes triangular spiral packing with a particle size of 3 mm to 5 mm. The pressure at the top of the column is 100 kPaA to 120 kPaA, and the temperature at the bottom is 95℃ to 105℃. (2) During the removal of light components, a light component removal tower is used, and the theoretical number of trays of the light component removal tower is controlled to be 20 to 30, the packing is 3mm to 5mm triangular spiral packing, the top pressure of the tower is 1.5kPaA to 2kPaA, the bottom temperature is 120℃ to 130℃, and the reflux ratio is (10 to 15):

1. (3) During the deweighting process, a deweighting tower is used, and the theoretical number of trays of the deweighting tower is controlled to be 20 to 30, the packing is 3mm to 5mm triangular spiral packing, the top pressure is 1.5kPaA to 2kPaA, the bottom temperature is 120℃ to 130℃, and the reflux ratio is 1: (15 to 20).

10. The (R)-citronellol separation method according to any one of claims 4 to 9, characterized in that, The (R)-citronellol separation device according to any one of claims 1 to 3 is used.

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