Method for producing material

Abstract

A method of producing a final matrix selected from the group consisting of esters, fatty acid esters, acids, alcohols, aldehydes, apocarotenoids and sesquiterpenes; wherein a suitable precursor fluid material is added to a rotating spiral bioreactor having a spiral channel coated with a biofilm; wherein the biofilm material comprises a material as a catalyst for converting the precursor material to the final matrix material; wherein the catalyst material is selected from the group consisting of: yeast, mould, fungus, bacterium or enzyme, preferably yeast, bacterium or enzyme, more preferably yeast; the method is characterized in that a spiral channel rotates for a period of time while a precursor fluid material is in the spiral channel coated with a biological membrane; the resulting final substrate is then removed.

Description

[0001] TECHNICAL FIELD This invention relates to a method for producing materials, and in particular a method for producing a final matrix selected from esters, fatty acid esters, acids, alcohols, aldehydes, apocarotenoids, and sesquiterpenes. BACKGROUND This invention relates to methods for producing materials, particularly methods for producing a final matrix selected from esters, fatty acid esters, acids, alcohols, aldehydes, apocarotenoids, and sesquiterpenes. These materials are commonly used as fragrance ingredients and are therefore high-value materials, despite the generally lack of pathways for their production. For example, 2-phenylethanol (2PE), which has a “rose-like” aroma, is an important molecule in the fragrance and flavoring industry. Conventional 2PE production methods involve extraction from the essential oils of many flowering plant species—most notably rose oil, which contains up to 60% 2PE. Although extraction is still practically used to obtain natural products, due to the high cost and poor scalability of the method, most 2PE production is now carried out via chemical synthesis from petrochemical feedstocks. While cheaper, 2PE production in this manner is non-renewable and unsustainable, and also uses carcinogenic precursors (e.g., benzene) as feedstocks. Alternatively, 2PE can be produced using established biotechnological methods via microbial fermentation or extracted from rose petals. However, conventional biotechnological methods result in low yields of 2-phenylethanol, often because concentrations exceeding 2 g / L are toxic to microorganisms. Furthermore, extracting oil from rose petals relies on using a large amount of land to grow roses, and typically requires more than 30,000 rose petals to produce less than 1 kilogram of oil, thus incurring extremely high costs.

[0003] Gao and Daugulis reported productivity results for a CSTR (continuous stirred tank reactor) using the yeast strain Kluyveromyces marxianus (CBS 600) with and without ISPR (in-situ product removal). The specific production rate was 0.05 g / L / h without product removal (typical in current industrial practice), compared to 0.43 g / L / h with product removal (Gao, f. and AJ Daugulis, 2009, Bioproduction of the Aroma Compound 2-Phenylethanol in a Solid-Liquid Two-Phase Partitioning Bioreactor System, Biotechnology and Bioengineering 104:2, 332-339).

[0004] Other such fragrance / aromatic molecules, particularly esters, fatty acid esters, alcohols, aldehydes, apocarotenoids, and sesquiterpenes, are similarly difficult to produce in the required quantities through inefficient industrial processes or due to the large amounts of land and other resources required to extract these substances from nature. The question remains: how can these materials be produced in a more efficient, renewable, and sustainable manner?

[0005] This problem can be overcome by the method of the present invention described herein. SUMMARY This invention relates to a method for producing a final matrix selected from esters, fatty acid esters, acids, alcohols, aldehydes, apocarotenoids, and sesquiterpenes; In this process, a suitable precursor fluid material is added to a rotating spiral bioreactor with spiral channels coated with a biofilm. Biomembrane materials include materials that serve as catalysts for converting precursor materials into final matrix materials; The catalyst material is selected from yeast, mold, fungus, bacteria or enzyme, preferably yeast, bacteria or enzyme, more preferably yeast; The feature is that the helical channel rotates for a period of time simultaneously with the precursor fluid material being coated with a biofilm within the helical channel; The resulting final matrix is ​​then removed.

[0007] An example has been given of using a rotating helical bioreactor to demonstrate that the final matrix material yield can be 15 times higher than that of conventional production processes using a continuous stirred tank reactor.

[0008] A further advantage is that the biofilm material containing the catalyst is not lost in the effluent, which occurs in conventional production processes using continuous stirred tank reactors.

[0009] Furthermore, compared to conventional production processes using continuous stirred tank reactors, much higher particle loading can be achieved in helical bioreactors, resulting in much higher final product yields.

[0010] Another advantage of helical bioreactors over conventional stirred tank continuous flow reactors includes increased productivity through precise control of mass transfer conditions, inherent reactor temperature uniformity control, and simultaneous removal of product substances. This also allows for easier and faster product removal, thus limiting any negative impact of the products on the system, such as if the final products poison the reaction or organism used.

[0011] The rotation of the helical channel offers numerous advantages. Two main advantages are the control of flow throughout the fluid's passage through the reactor, and primarily due to this control, the ability to theoretically capture flow rate, mass transfer, and reaction characteristics. The first advantage allows the suspension to remain homogeneous during injection, separates the fluid phase into parallel flows of constant thickness along the channel length, controls the interface between fluid phases, balances the feed liquid and gas to apply gas species concentrations, and facilitates efficient heat transfer in the gas throughout the shell surrounding the rotating assembly (thus allowing for uniform temperature application within the reactor and maintaining optimal mass transfer conditions throughout). The second advantage allows for the avoidance of trial-and-error design and enables faster reactor optimization.

[0012] Preferably, the biofilm coating of the spiral channel is carried out in the following manner: a) i) Loading a suspension containing particles of catalyst material into a spiral channel; and ii) Rotate the spiral channel for a period of time while the suspension is inside the spiral channel; iii) Optionally, but preferably, the supernatant is removed; or b) i) While the helical channel rotates, a suspension containing particles of catalyst material is loaded into the helical channel; and ii) Continue the rotation of the spiral channel for a period of time; iii) Optionally, but preferably, the supernatant is removed.

[0013] Preferably, the final matrix is ​​selected from ethyl acetate, isoamyl acetate, isoamyl alcohol, benzaldehyde, 2-phenylethyl acetate, 2-phenylethyl butyrate, phenylethyl propionate, methyl jasmonic acid, 2-phenylethanol, C2-C6 carboxylic acid, hexanal, octanal, decanal, dodecaldehyde, vanillin, apocarotenoids (preferably α- and β-ionones), and sesquiterpenes. More preferably, the final matrix is ​​selected from ethyl acetate, isoamyl acetate, isoamyl alcohol, benzaldehyde, 2-phenylethyl acetate, 2-phenylethyl butyrate, phenylethyl propionate, methyl jasmonic acid, 2-phenylethanol, C2-C6 carboxylic acid, hexanal, octanal, decanal, dodecaldehyde, vanillin, α- and β-ionones, and sesquiterpenes. More preferably, the final matrix is ​​selected from ethyl acetate, isoamyl acetate, isoamyl alcohol, benzaldehyde, 2-phenylethyl acetate, 2-phenylethyl butyrate, phenylethyl propionate, methyl jasmonic acid, and 2-phenylethanol. Most preferably, 2-phenylethanol.

[0014] Preferably, the yeast material is selected from *Corydalis lucida* (Portunus lucida). Clavispora lusitaniae Kluyveromyces martensii, species of the genus Kluyveromyces ( Kluyveromyces sp. Dobkluyces yeast ( Kluyveromyces dobzhanskii ), brewer's yeast (Saccharomyces cerevisae ), Delbbu's spore-forming yeast ( Torulaspora delbruekii ), non-yeast yeasts from Durango agave, brewer's yeast ( Saccharomyces cerevisiae ), Saccharomyces cerevisiae ( Schizosaccharomyces pombe ), Little Red Yeast ( Rhodotorula minuta ), species of the genus Pistil ( Pithomyces sp. ), Beauveria bassiana ( Beauveria bassiana ), Crimson Fungi ( Pycnoporus cinnabarinus Aspergillus niger ( ) Aspergillus niger ) and Yersinia lipophila ( Yarrowia lipolytica Preferably, the bacteria are selected from Escherichia coli (Escherichia coli). E. Coli Streptomyces ( Streptomyces ), Pseudomonas ( Pseudomonas Acetobacter () Acetobacter Staphylococcus aureus ( Gluconobacter ) and lactic acid streptococci ( Lactic streptococci Preferably, the enzyme is selected from alcohol dehydrogenase (ADH), flavin-dependent alcohol oxidase, copper radical oxidase, lipase, and acetyltransferase. Attached Figure Description

[0015] Figure 1 It is a comparison of usage Figures 3-5 The graph shows the rate of 2-phenylethanol production using a rotating helical bioreactor compared to that using CSTRs (comparison). Figure 2A -B shows a plan view of the spiral channel during use, where Figure 2B Showing Figure 2A A magnified view of a portion; Figure 3 This is a perspective view of the rotating helical bioreactor used in this invention, with the positioned side not shown to show the reactor bed; Figure 4A and Figure 4B This is a photograph of the rotating helical bioreactor used in this invention; Figure 5 This is a cross-sectional view of the rotating helical bioreactor used in this invention, wherein the lines indicate the liquid flow path; Figure 6 This is a cross-sectional view of the rotating helical bioreactor used in this invention, which has a six-helical-channel reactor bed and is configured to allow a second liquid to flow in a countercurrent manner (e.g., for product extraction). Figure 7A -C displays something similar to Figure 6 The diagram shows a cross-sectional view of the reactor. Figure 8 yes Figure 7A -C is a photograph of the rotary helical bioreactor used in the present invention, which is configured for an extraction flow with six helical channels; Figure 9A -F shows a schematic cross-sectional view of the rotating helical bioreactor used in this invention, illustrating the various fluid networks; Figure 10 A schematic cross-sectional view of the rotating helical bioreactor used in this invention is shown, illustrating a fluid network with three reactor beds; Figure 11 Shown in Figures 3-5 The diagram shows a plan view of the bed of watermelon seed powder and deionized water flow in the spiral channel of the rotating spiral bioreactor used in this invention. Figure 12 Showing the use of, for example Figures 3-5 The graph shown illustrates the effect of bed thickness on particle loading in the initial suspension of watermelon seed powder in a rotating helical bioreactor. Figure 13A -B shows the following plan view: A) Growth of the initial bed of Max Kluyveromyces cells, B) In such a plan view Figures 3-5 The growth of Max Kluwer yeast cells in the spiral channel of the rotating spiral bioreactor shown at the end of an 8-hour production run; Figure 14 Shown in Figures 3-5 A plan view of oxygen generated by a bed of watermelon seed powder in the spiral channel of a rotating spiral bioreactor. DETAILED DESCRIPTION Unless otherwise stated, the indefinite article “a” or “an” and its corresponding definite article “the” as used herein mean at least one or more. As described herein, any preferred subject matter may be explicitly combined with other preferred subject matter.

[0017] Other advantages of using a spiral bioreactor to produce the selected final substrate Unrestricted by theory, using a rotary screw reactor offers several advantages. Two main advantages of a rotary screw reactor are controllability and predictability.

[0018] Regarding control, the rotary helical reactor allows for control over the manner of physical phase contact. For example, the thickness of the particle bed can be controlled independently of the fluid flow rate. Therefore, it is possible to control the mass transfer rate, particularly optimizing it during the period of phase contact (i.e., during the reaction process).

[0019] Rotary helical reactors can handle high particle volume fractions. This is achieved by preventing particles from distributing into the fluid or forcing the supply fluid through areas occupied by particles. Instead, the particles are arranged along one side of the channel in the bed, i.e., in a layer, and the fluid flows adjacent to rather than through the bed. Mass transfer between the fluid flow and the bed is achieved through diffusion. To control the mass transfer rate, the layer thickness can be controlled. The layer thickness can depend on factors such as the reaction rate and the diffusion rate. A solid layer thickness of a few millimeters has been found to be suitable. Depending on the required flow width of the supply bed, particle volume fractions up to 20% have been achieved, and it is thought that particle volume fractions up to 40% can be achieved.

[0020] Regarding predictability, phase behavior is more predictable due to the relatively simple phase configuration within the spiral reactor. This enhanced predictability allows users to reliably design new devices and facilitates system optimization of devices associated with specific reaction systems (e.g., particle type and / or the reaction to be carried out).

[0021] Compared to conventional CSTRs, the advantages of helical reactors associated with improved control and predictability include increased productivity. This can be achieved through precise control of mass transfer conditions, inherent control of reactor temperature uniformity, and simultaneous removal of product substances. Simultaneous removal of product substances also allows for easier and faster product removal and can limit any negative impacts of the product on the system, such as if the product's presence affects the equilibrium of product formation or otherwise inhibits the reaction (e.g., by denaturing enzymes and / or poisoning the organisms used in the reaction).

[0022] In contrast, CSTRs encounter problems as the particle volume fraction increases. In the literature and likely in industrial practice, the volume fraction is kept below 10%. For mass transfer purposes, the particles are in suspension, and with a volume fraction approaching 10%, the typical space between particles is less than one particle diameter, which increases the apparent viscosity of the suspension. Therefore, as the particle volume fraction increases, the effective viscosity of the reaction mixture also increases accordingly. Consequently, the ability to introduce further reagent supply decreases, and the mass transfer rate decreases.

[0023] Therefore, compared to CSTRs, rotary helical reactors can achieve higher particle volume fractions and more efficient mass transfer of reagents, nutrients, products, and waste. Compared to CSTRs, using rotary helical channel reactors yields orders of magnitude improvements.

[0024] In addition to these advantages, the benefits associated with reactions in the absence of particles can be further enhanced by the additional controlled operation of the fluid phase, namely gas operation and liquid extraction.

[0025] Another advantage of spiral reactors is less disruption to heavier phases (i.e., denser, e.g., solid phases). For example, in reactions where lighter phases (e.g., gases released during the reaction) must be separated from other phases, the reactor design allows the lighter phase to flow along the inner wall of the spiral channel. This does not significantly disturb the heavier phase on the relative (outer) wall of the spiral channel. In CSTRs, disruption is significant. Therefore, phase isolation becomes possible. This is crucial because gas generation typically increases disruption to heavier phases (e.g., solid phases). This provides particular benefit when the phase is used for extraction and / or the phase needs to be separated.

[0026] Description of an example of a rotating spiral bioreactor A rotating helical bioreactor should be understood as a reactor designed for biotransformation reactions that can rotate during use. "Rotating" and "rotatable" are interchangeable.

[0027] The description herein is a description of an example of a rotating helical bioreactor. This illustrative rotating helical bioreactor is a preferred example of a rotating helical bioreactor that can be used in the methods of the present invention; however, it is merely an example and is intended as a non-limiting example of such a rotating helical bioreactor for use in the methods of the present invention. Any language indicating the nature of the rotating helical bioreactor described herein is intended only to relate to this specific exemplary embodiment of the rotating helical bioreactor and not to the nature of the more general method claimed, which can use any helical bioreactor that provides a channel that is spiraled while a precursor fluid material is situated within a biofilm-coated helical channel.

[0028] A non-limiting example of a rotating helical bioreactor is described herein, and particularly in the following description with reference to the accompanying drawings.

[0029] Preferably, the rotating helical bioreactor comprises: At least one reactor bed comprising a helical channel, wherein the helical channel extends from an inner end to an outer end, and wherein the reactor comprises: • At least one receiver unit comprising an annular reservoir, wherein the receiver unit rotates together with the reactor bed or each reactor bed, and wherein the receiver unit is configured to receive liquid from a non-rotating source and transfer the liquid to a helical channel; and / or • At least one output unit comprising an open-ended conduit, wherein the conduit rotates with the reactor bed or each reactor bed, and wherein the conduit is configured to form a liquid jet from a helical channel such that, in use, the jet can be collected by a non-rotating annular collector.

[0030] The reactor may include at least one receiver unit and at least one output unit, for example, one or two receiver units and one or two output units. Preferably, the reactor has two receiver units and two output units.

[0031] By using one or more receiver units and / or output units to introduce one or more liquids into and / or unload them from a rotatable spiral reactor, the spiral reactor is simpler to use (and therefore more readily available to obtain the benefits of the spiral reactor) than a conventional rotatable spiral reactor, requires less maintenance, and has lower energy requirements.

[0032] It should be understood that a reactor may include one or more reactor beds, one or more helical channels, one or more receiver units, and one or more output units. While this disclosure may be made explicitly for each of these components, it is also intended to relate to more than one of each of these components, e.g., all of them, unless otherwise incompatible. It should be understood that the number of helical channels and / or reactor beds may be increased to increase the reactor's production capacity. It should be understood that the number of receiver units and output units may correspond to the amount of liquid phase the reactor is configured to process during use.

[0033] The rotatable body may include components in the reactor intended to rotate, such as the receiver unit and / or output unit or each receiver unit and / or output unit, the reactor bed or each reactor bed, the helical channel or each helical channel, and optional components, such as devices for controlling liquid pressure. The rotatable body may, for example, be rotatably coupled to a fixed frame via bearings.

[0034] The rotatable body may include a shaft, such as an elongated shaft. The (intended) axis of rotation may be centered on the axis of the shaft. The reactor components may be coaxial with each other, i.e., configured to rotate about substantially the same axis. For example, the (or each) receiver unit, spiral channel, reactor bed, and output unit may be coaxial with each other and / or with the shaft.

[0035] Technicians will understand that the acceleration on the reactor and the fluid therein depends on the radius of the apparatus and the rotational speed. The rotatable body and / or rotatable assembly of the reactor can be configured to withstand accelerations of 50 g or greater, or 100 g or greater, such as 400 g or greater, or 500 g or greater, such as 1000 g or greater (e.g., 100,000 g or less, such as 10,000 g or less, or 50 g to 100,000 g). The rotatable body and / or rotatable assembly of the reactor can be configured to withstand rotations at speeds of 200 RPM or higher, such as 300 RPM or higher, such as 400 RPM or higher, such as 2000 RPM or higher (e.g., 100,000 RPM or lower, such as 20,000 RPM or lower, or 200 RPM to 100,000 RPM).

[0036] Receiver unit The reactor may include at least one receiver unit. The receiver unit, or each receiver unit, includes an annular reservoir. The receiver unit, or each receiver unit, rotates together with (i.e., in sync with) the reactor bed. Therefore, in use, the receiver unit can rotate in sync with the helical channel / reactor bed, or each helical channel / reactor bed. The receiver unit is configured to receive liquid from a non-rotating source. The receiver unit is configured to transfer the liquid into the helical channel (i.e., into one end of the helical channel). The liquid can then flow through the helical channel.

[0037] The annular reservoir may include annular sidewalls. The annular sidewalls are preferably substantially centered on the (intended) axis of rotation. The annular sidewalls preferably extend along and around (i.e., always around) the (intended) axis of rotation.

[0038] The annular reservoir may have a base (e.g., a platform). The base is preferably generally perpendicular to the (intended) axis of rotation. The reservoir may be fed by a liquid flow from the base.

[0039] The receiver unit can have a substantially U-shaped cross-section, comprising a base and annular sidewalls. The receiver unit can ideally be configured to prevent liquid from escaping before entering the helical channel.

[0040] The delivery unit configured to supply liquid to the receiver unit may be (or may not be) part of the reactor. The delivery unit may include means for controlling the liquid flow rate (e.g., a syringe pump) and / or conduits arranged to supply liquid to the receiver unit.

[0041] Liquid entering the receiver unit (e.g., via a conduit or flow) typically first contacts the base of the receiver unit. Preferably, the base is circular. Preferably, the receiver unit (e.g., annular reservoir) is substantially rotationally symmetric about an axis of (intended) rotation.

[0042] Upon contact with the receiver unit, the liquid accelerates towards the annular sidewall of the annular reservoir due to rotation. The degree of acceleration towards the annular sidewall depends on the rotational rate of the receiver unit and / or the liquid flow rate. Those skilled in the art will understand that, in the case of initial contact with the base (platform), it is possible to calculate the depth of the liquid layer at the base as a function of radial position, and the average residence time of the liquid at the base before reaching the sidewall. Transfer of heat and / or mass (of gaseous matter) can occur during the time the liquid is in contact with the receiver unit. The rate of this transfer can depend on the flow rate, rotational rate, and platform arrangement. This can be controlled by adjusting the gas composition on the platform to allow the feed liquid to be saturated with gaseous matter and to ensure that the liquid entering the helical channel is at reactor temperature.

[0043] As the liquid is laterally accelerated due to rotation, the annular reservoir prevents liquid from escaping from the receiver unit. The cross-section of each side of the annular reservoir can be substantially J-shaped, C-shaped, I-shaped, or L-shaped. Preferably, each side of the annular reservoir has a substantially J-shaped cross-section. Preferably, the sidewalls are integral with the base.

[0044] The annular reservoir may include a top (e.g., a lid) to help prevent liquid from leaving the annular reservoir once it has entered and / or to prevent contaminants from entering the annular reservoir. The top may have an orifice (e.g., an annular orifice) to allow liquid to enter as the annular reservoir rotates. Preferably, the top is integral with the sidewalls of the annular reservoir.

[0045] Technicians will understand that the inner radius of the annular container edge can ideally be small enough that the receiver does not overflow at any stage or under any normal operating conditions. Higher flow rates ideally require a more radially arranged edge in the design. Gas operations may require increased receiver capacity because gas may exit the reactor through the receiver unit. Furthermore, if pulsating pumping (characteristic of positive displacement pumps) is used to introduce liquid into the receiver unit, the reservoir capacity can be increased to mitigate the velocity variations flowing into the helical channel.

[0046] The receiver unit is configured to receive liquid from a non-rotating source. Therefore, the receiver unit may have an annular opening to allow liquid to be introduced from the non-rotating source. This allows a static conduit to load liquid into the receiver unit. Preferably, the annular opening is centered on the axis of rotation and extends around the axis of rotation in a plane perpendicular to the axis of rotation.

[0047] The receiver unit is configured to transfer liquid into a spiral channel. The receiver unit has one or more outlets (e.g., conduits) that connect the inner surface of the reservoir to the spiral channel or one end of each spiral channel. This allows liquid received by the receiver unit to be conveyed into the spiral channel. Preferably, the outlets of the receiver unit are located on the base (e.g., outside the outer edge of the base) and / or on the annular sidewall (e.g., at the edge of the annular sidewall contacting the base). The reservoir may have a separate outlet for each spiral channel; for example, a receiver unit for a 10-spiral channel stack may have 10 outlet connection passages equidistantly spaced around the annular reservoir. A single connection passage from a shared reservoir is considered to allow for optimal space utilization and to accommodate necessary structural elements.

[0048] The receiver unit is configured to receive liquid and to transfer the liquid to a spiral channel. The receiver unit may be configured to transfer the liquid to the inner or outer end of the spiral channel. Preferably, at least one receiver unit of the reactor is configured to transfer the liquid to the inner end of the spiral channel. Preferably, the reactor has two or more receiver units, wherein a first receiver unit is configured to transfer the liquid to the inner end of the spiral channel, and wherein a second receiver unit is configured to transfer the liquid to the outer end of the spiral channel.

[0049] Reactor bed The reactor comprises at least one reactor bed containing helical channels. In practice, the reactor bed is the body defining one or more helical channels, for example, a conduit. It should be understood that more than one helical channel is typically arranged in different planes relative to each other. The helical channels generally do not intersect each other; in other words, they are typically independent of each other. The reactor may comprise multiple stacked reactor beds.

[0050] The reactor comprises one or more, such as two or more, or four or more reactor beds. The reactor may include 200 or fewer, such as 100 or fewer, or 50 or fewer, such as 40 or fewer, or 20 or fewer. The reactor may include 1 to 50 reactor beds, such as 2 to 40, or 2 to 20. The reactor bed, or each reactor bed, includes one or more helical channels, such as two or more, or four or more. The reactor bed, or each reactor bed, may include 50 or fewer helical channels, such as 40 or fewer, or 20 or fewer. The reactor may include 1 to 50 helical channels, such as 2 to 40, or 2 to 20.

[0051] Each reactor bed may have a fluid connection for the inner end of each helical channel within the reactor bed. Each reactor bed may have a fluid connection for the inner ends of some or all of its contained helical channels. Each reactor bed may have a fluid connection for the outer end of each helical channel within the reactor bed. Each reactor bed may have a fluid connection for the outer ends of some or all of its contained helical channels.

[0052] In reactors comprising more than one helical channel, the reactor may include conduits connecting the inner or outer ends of the helical channels. These conduits ensure that the feed liquid is evenly distributed to each helical channel. There may be one such conduit connected to the inner end and another connected to the outer end. Such conduits may be referred to as columns. In use, such conduits may contain a column of static liquid. The conduit connected to the inner end of the helical channel is used for the lower dense phase in use. The conduit connected to the outer end of the helical channel is used for the higher dense phase.

[0053] The reactor bed can be substantially cylindrical. The reactor bed may include conduits to fluidly connect the helical channel or each helical channel to the receiver unit, the inner or outer end of the conduit or each connected helical channel, the output unit, and / or the weir.

[0054] Each helical channel can be sealed (across the entire cross-section of the helical channel) to prevent fluid from escaping the helical channel, especially under rotation. The reactor may include a cover for the reactor bed or each reactor bed to seal the helical channel or each helical channel. One reactor bed may serve as a cover for a second reactor bed (of the helical channels). The cover for the helical channels may include orifices and / or conduits configured for inlet and / or outlet flow.

[0055] Spiral channel Rotary reactors offer particular advantages when used with multiphase systems, such as those where phases are in contact within a rotating helical channel. The rotating helical channel allows solid phases (e.g., cells, such as biofilms) to be stabilized by centrifugation. The solid phase can be continuously supplied with a sustaining medium, such as a reagent, flowing over the liquid phase.

[0056] The reactor may include one or more helical channels. Preferably, the rotary helical reactor includes multiple helical channels. The reactor may include two or more helical channels, or four or more, or ten or more helical channels. The reactor may include 100 or fewer helical channels, for example, 50 or fewer helical channels, or 20 or fewer helical channels. The reactor may include 1 to 100 helical channels, for example, 2 to 100 helical channels, or 4 to 50 helical channels.

[0057] In reactors with two or more helical channels, the helical channels are preferably connected in parallel. Preferably, the two or more helical channels have a common interconnection at one or both ends of each channel to provide equal flow to each individual helical channel. Preferably, the same ends (i.e., the inner and / or outer ends) of each helical channel are connected together as a common connection. Preferably, the common connection between the helical channels is equidistant from the center and / or axis of rotation of the helical channels (at the point where it connects to each helical channel). Preferably, the common connection between the helical channels is equidistant from the axis of rotation of the helical channels along the path between its connections to each helical channel.

[0058] Preferably, the reactor includes means for controlling the liquid pressure in the helical channel (e.g., the pressure at the outer end of the helical channel). In examples of helical bioreactors used in the method of the present invention, the liquid level in the reservoir can be self-regulating by using means for controlling the liquid pressure. The means for controlling the liquid pressure in the helical channel is particularly useful for liquid flows to an output unit containing an open conduit at the end, to prevent liquid from flowing directly through the reactor.

[0059] The device for controlling liquid pressure is preferably a "weir" fluidly connected to the end of a helical channel, wherein the top of the weir is positioned closer to the center of the (in use) rotating shaft and / or the helix (defined by the helical channel) than the end of the helical channel to which the weir is connected. Preferably, the device (e.g., the weir) is fluidly connected to the outer end of the helical channel. The reactor may include a first device (e.g., a weir) connected to the inner end of the helical channel and a second device (e.g., a weir) connected to the outer end of the helical channel. Preferably, the top of the weir is positioned closer to the center of the (in use) rotating shaft and / or the helix than the inner end of the helical channel. Preferably, the weir / weir is fluidly connected to the end of the helical channel using a conduit. The conduit is arranged to guide liquid flowing out of the outer end of the helical channel to the device / weir, i.e., to guide liquid flowing out of the outer end to the center of the (in use) rotating shaft and / or the helix. It should be understood that the liquid flow rate is proportional to the liquid flow rate entering the receiver unit, which can be controlled by an external pump and / or a pressure regulator of the delivery unit. In the case of gas flow, an external pressure source is required to generate a difference relative to the shell pressure or other reference pressure.

[0060] The weir, or each weir, can establish a fixed hydrodynamic reference (combining pressure, gravity, and centrifugal energy) with respect to the radial level of the liquid or gas in the receiver unit. The weir can force the reservoir level to exceed this reference by an amount proportional to the frictional losses in the helical flow. The liquid velocity is determined by the rate at which the liquid is fed into the receiver unit. The radial position of the weir (i.e., its distance from the axis of rotation) and the incoming liquid velocity determine the liquid level in the annular receiver (which is developed to drive forced flow). In the presence of multiple liquid phases and multiple phases, it should be understood that the radial position of the weir and the incoming liquid velocity determine the liquid level in the annular receiver (which is developed to drive forced flow).

[0061] Those skilled in the art will understand that, in the case of liquid-liquid flow, the radial position of the weir can be ideally configured such that the separated phases flow out of the reactor separately, i.e., the heavy liquid phase does not flow out of the light phase outlet, and the light phase does not flow out of the dense phase outlet.

[0062] The benefits associated with introducing and / or unloading liquids from a rotary screw reactor allow users to more easily access the benefits associated with rotary screw reactors, such as controllability. The screw reactor promotes phase retention in distinct continuous phases, thus avoiding the formation of slugs and / or foaming and / or emulsification of individual phases. Flow and interface shapes are simple and predictable.

[0063] Spiral reactors can be used for in-situ product removal, demonstrating improved efficiency compared to conventional reactors.

[0064] A (separate / separate) device (e.g., a weir) may be present to control the liquid pressure between each helical channel and the common connection. Preferably, a device is present to control the liquid pressure between the outer end of each helical channel and the outer common connection. Once past the device for controlling the liquid pressure (e.g., a weir), individual outlet flows are preferably merged to use a common output unit, but they can use separate output units. Additionally or alternatively, a device (e.g., a weir) may be present to control the liquid pressure between the inner end of each helical channel and the inner common connection, particularly in cases where a lower-density liquid phase flows countercurrently with a higher-density liquid phase.

[0065] A device (e.g., a weir) for controlling the liquid pressure of each common connection, fluidly connected between the common connection and the output unit, may be present. Therefore, more preferably, a device (e.g., a weir) for controlling the liquid pressure between the common connection and the output unit (e.g., fluidly connected between the common connection and the output unit) may be present. A device for controlling the liquid pressure between each helical channel and the common connection may not be present. More preferably, a device (e.g., a weir) for controlling the liquid pressure between the outer common connection and the output unit (e.g., fluidly connected therein) and a device (e.g., a weir) for controlling the liquid pressure between the inner common connection and the output unit (e.g., fluidly connected therein) may be present. A device for controlling the liquid pressure between each helical channel and the common connection may not be present.

[0066] Preferably, two or more helical channels have a common connection at both ends of the channel, which can be referred to as the "inner common connection" and the "outer common connection" for the inner and outer ends of the helical channel, respectively. In a system in which fluid flows through the helical channel in only one direction, the inner common connection is usually located at the inlet of the helical channel, and the outer common connection is usually located at the outlet of the helical channel. However, it is possible that the inner common connection is the output of the helical channel and the outer common connection is the input of the helical channel. In a system in which fluid flows through the helical channel in two directions (countercurrent), the inner common connection and the outer common connection each act as the input and output of the helical channel, respectively.

[0067] In a system with a single liquid phase flow, the common inlet condition is a single receiver reservoir free surface hydraulic reference, and the common outlet condition is a single outlet weir located exactly in the same radial position to provide a common hydraulic reference. In the case of two immiscible liquid phases (co-current or counter-current), four different common connections may be involved: two different reservoir levels and two sets of outlet weir locations, although the common input of one phase can be shared with the common output of another phase, and vice versa.

[0068] The center of the helix can be called the origin of the helix. The helical channel, or each helical channel, can be centered on an axis of rotation (i.e., the origin coaxial with it), around which the reactor bed rotates in use. In the case of a reactor having two or more helical channels, the helical channels preferably have the same axis of rotation (i.e., coaxial with each other).

[0069] In the case of two or more helical channels and / or reactor beds, each helical channel and / or reactor bed may be offset relative to each other in azimuth (e.g., around an axis of rotation) while remaining coaxial with each other. The offset can be configured to rotatably balance the helical channels and / or reactor beds. For example, the helical channels may be offset relative to each other by 360 / n degrees in azimuth (e.g., around an axis of rotation), where n represents the number of helical channels.

[0070] The helical channel can be substantially flat (e.g., flat), meaning it extends in two dimensions, i.e., in a plane. The helix can extend beyond the plane, but preferably it does not. The helix can extend to the axis of rotation, but typically the helix does not.

[0071] The spiral channel can extend one or more full turns, such as two or more, preferably four or more, or six or more, such as seven or more, or eight or more full turns. The spiral channel can extend 100 or fewer full turns, such as 80 or fewer, or 60 or fewer, such as 40 or fewer, preferably 20 or fewer, for example, 15 or fewer, or 12 or fewer, or 10 or fewer, such as 9 or fewer, or 8 or fewer full turns. The spiral channel can extend from 1 to 100 full turns, such as 2 to 40 full turns, or 4 to 20 full turns.

[0072] The minimum inner diameter of a spiral, measured as the minimum distance along the spiral axis from the center of one side of the spiral channel to the center of the other side (i.e., twice the minimum radius of the inner end of the spiral channel), can be 1 cm or larger, such as 5 cm or larger, or 8 cm or larger, or 10 cm or larger. The minimum inner diameter of a spiral can be 2 meters or smaller, such as 1 meter or smaller, or 50 cm or smaller, for example, 40 cm or smaller, or 30 cm or smaller. The minimum inner diameter of a spiral can range from 1 cm to 2 meters, such as 5 cm to 1 meter, or 8 cm to 40 cm.

[0073] The maximum outer diameter of a helix, measured as the maximum distance from the center of one side of the helical channel to the center of the other side through the helical axis (i.e., twice the maximum radius of the outer end of the helical channel), can be 2 cm or greater, or 5 cm or greater, such as 10 cm or greater, or 20 cm or greater, or 30 cm or greater, or 70 cm or greater, or 1 meter or greater, such as 2 meters or greater. The maximum outer diameter of a helix can be 10 meters or less, such as 5 meters or less, or 2 meters or less, such as 1 meter or less, for example, 80 cm or less, or 60 cm or less, such as 40 cm or less. The maximum outer diameter of a helix can be from 10 cm to 10 meters, for example, from 20 cm to 1 meter, or from 20 cm to 80 cm.

[0074] The inner end of the helical channel is closer to the (intended) axis of rotation than the outer end. Preferably, the radius of the helical channel from the (intended) axis of rotation increases progressively from the inner end to the outer end. The increase in radius is preferably uniform along the length of the helical channel. Preferably, the helical channel has a smooth wall surface (i.e., neither uneven nor rough).

[0075] The type of helix can be chosen such that a substantially constant centrifugal force acts on the fluids as one or more fluids pass through the helical channel. This is important for ensuring constant contact conditions along the entire channel. These benefits are particularly evident in multiphase (e.g., multifluid phase) systems, where consistency of contact conditions along the length of the helical channel can be crucial. The helical channel can be in the form of an Archimedean spiral, a hyperbolic spiral, or a Fermat spiral. Preferably, the spiral is an Archimedean spiral, which provides a constant centrifugal force on the phases along the helical channel.

[0076] The channel can have any suitable depth. The channel depth can be determined as the cross-sectional dimension at any location along the channel length parallel to the intended axis of rotation and / or perpendicular to the helical plane. The channel depth can be 0.01 mm or greater, preferably 0.05 mm or greater, or 0.08 mm or greater, such as 0.1 mm or greater, such as 0.5 mm or greater, or 1 mm or greater, such as 1.3 mm or greater. The depth can be 50 mm or less, or 20 mm or less, preferably 10 mm or less, or 8 mm or less, such as 5 mm or less, for example, 3 mm or less, or 2 mm or less. The depth can range from 0.01 mm to 50 mm, such as 0.05 mm to 10 mm, preferably 0.05 mm to 5 mm.

[0077] The channel can have any suitable width. The width of the channel can be determined as the cross-sectional dimension at any location along the length of the channel that is perpendicular to the intended axis of rotation (and / or parallel to the helical plane). The width of the channel can be 0.01 mm or greater, such as 0.05 mm or greater, preferably 0.1 mm or greater, or 0.2 mm or greater, such as 0.5 mm or greater, or 1 mm or greater, such as 1.3 mm or greater. The width can be 50 mm or less, or 20 mm or less, such as 10 mm or less, preferably 5 mm or less, for example, 3 mm or less, or 2 mm or less, such as 1.8 mm or less, or 1.2 mm or less, or 1 mm or less. The width can be from 0.01 mm to 50 mm, such as 0.05 mm to 10 mm, preferably 0.1 mm to 5 mm.

[0078] The cross-sectional shape of the channel perpendicular to the helix length can be any suitable shape. Preferably, the shape is circular, elliptical, or rectangular (e.g., square, optionally with rounded or non-circular corners). Those skilled in the art will understand that when a solid phase is used in the reactor, the thickness of the solid phase at each location is determined by the height of the suspension directly above it (in the radial direction). Thus, for example, a circular cross-section results in the thickest solid phase at the center of the outer wall, with the thickness decreasing away from this center. Furthermore, particles settling on the inclined portions of the outer wall may slide towards the center. This corresponds to relatively poor channel utilization. Preferably, the channel shape is rectangular, especially when a solid phase is present within the reactor, thereby allowing the formation of a solid phase of uniform thickness.

[0079] Controlling the flow rate of the fluid The flow rate of fluid through the helical channel can be varied depending on the action taken. For example, a high flow rate may be preferred during suspension injection and / or cleaning.

[0080] The reactor may include means for controlling the flow rate of liquid through the spiral channel.

[0081] Technicians will understand that the flow rate through the reactor (during its steady state, when the channels are filled) depends on the rate at which fluid is introduced into the helical channels. Therefore, controlling the flow rate requires controlling the rate at which fluid is introduced into the helical channels. Increasing the rate at which fluid is introduced into the helical channels increases the flow rate of fluid through the helical channels. The liquid level within the helical channels can be controlled by a device that controls the pressure within the helical channels (e.g., a weir), thereby establishing a reference that is also present in the reservoir of the receiver unit. Therefore, similar to a U-bend in conventional piping applications, introducing liquid into the receiver unit at a faster rate can increase the flow rate through the helical channels. This device may include, for example, an external pump (e.g., for liquids) or a pressure regulator (e.g., for gases) to control the rate at which fluid flows into the receiver unit or through the gas passage.

[0082] In the presence of two immiscible liquid phases in a helical channel stack, providing a quasi-static liquid column of either the heavy or light phase can be used to ensure a uniform distribution of fluid from the receiver unit. These are described above as being interconnected.

[0083] Controlling the pressure in the spiral channel The reactor may include one or more means for controlling the pressure within the helical channel, such as a weir. Preferably, the means for controlling the pressure within the helical channel is (fluidically) connected to the outer end of the helical channel. The reactor may additionally or optionally include means for controlling the pressure within the helical channel (fluidly) connected to the inner end of the helical channel.

[0084] The device for controlling liquid pressure can be a conduit with a cross-sectional area smaller than that of the helical channel, wherein the conduit is fluidly connected to the inner end and / or outer end of the helical channel. The cross-sectional area of ​​the helical channel can be smaller than the cross-sectional area of ​​the conduit connecting the helical channel to one or both of the receiver and output units.

[0085] Preferably, the device, or each device, is a weir. The weir is fluidly connected to the (inner / outer) end of the helical channel. The top of the weir is positioned closer to the (in use) axis of rotation and / or the center of the helix. The weir makes the reactor somewhat similar to a U-bend in a conventional piping application, where the liquid level on one side is substantially the same as the liquid level on the other side when no liquid is added. In a U-bend, gravity acts on the liquid. In the reactor, rotation increases the centrifugal force acting on the liquid, and the weir can be used in combination with the rotational speed to control the pressure within the helical channel.

[0086] The reactor may include a conduit arranged to direct liquid flowing out of the helical channel toward the center of the rotating shaft and / or the helix in use. The conduit may fluidly connect the outer end of the helical channel to a weir. Preferably, the conduit is substantially parallel to the plane of the helical channel. Preferably, the conduit is substantially parallel to the radius of the (intended) rotating shaft, i.e., perpendicular to the (intended) rotating shaft.

[0087] The weirs discussed here (i.e., the top of the weir) typically establish one or more hydrodynamic references, with the reservoir water level supplying the helical channel. A fixed hydrodynamic reference combining pressure, gravity, and centrifugal energy allows the fluid within the reactor to withstand high forces caused by the rotation of the helical channel in use.

[0088] When the reactor includes more than one helical channel, each helical channel may have a corresponding device for controlling the pressure inside the helical channel, or two or more helical channels may share a device for controlling the pressure inside these helical channels.

[0089] In the case of two or more phases, a device with a higher density phase (e.g., a weir) can be located further away from the axis of rotation compared to a lower density phase.

[0090] The common connection discussed above can be used to connect each helical channel together, so that one or two devices for controlling the pressure within the helical channel (e.g., a weir) can be used for multiple helical channels.

[0091] Output unit The reactor includes at least one output unit comprising an open-ended conduit. The open-ended conduit rotates with the reactor bed or each reactor bed. The conduit is configured to form a liquid jet from a helical channel such that, in use, the jet can be collected by an annular collector.

[0092] Liquid typically enters the output unit from the helical channel or each helical channel through conduits. The output unit is fluidly connected to the inner or outer end of the helical channel or each helical channel, for example, by means of a device (e.g., a weir) for controlling the liquid pressure in the helical channel or each helical channel. In use, the output unit rotates in sync with the helical channel.

[0093] The output unit, or each output unit, includes an open-ended conduit that rotates with the reactor bed. The conduit can be, for example, a tube, pipe, or channel. The conduit may terminate at a nozzle or overflow outlet. The open-ended conduit is arranged to form a liquid jet from the end of the helical channel, such that, in use, the jet can be received by an annular collector.

[0094] The open end of the conduit is typically substantially perpendicular to and away from the axis of rotation, such that during use, the jet formed from the open end of the conduit travels away from the axis of rotation to be captured by the inner surface of the annular collector. The open end of the conduit can be at an angle of ±50° or less to the direction perpendicular to the axis of rotation, such as ±45° or less, preferably ±40° or less, or ±35° or less, such as ±30° or less, such as ±25° or less, such as ±15° or less (i.e., in the case where the axis of rotation is vertical rather than horizontal). This angle can be ±5° or greater, such as ±15° or greater, such as ±20° or greater, or ±25° or greater. This angle can be ±5° to ±50°, such as ±45° to ±15°, or ±40° to ±20°. This allows for higher jet elasticity and reduces jet breakup and / or atomization, thereby improving reactor efficiency. Generally, the smaller the angle, the thinner the liquid jet, which makes the jet more prone to breakup before collection. The actual flight angle is the vector sum of the angles under the tangential components of the average outflow velocity and rotational velocity (r). ω). Generally, the latter is much larger than the former, so the overflow angle has a relatively small impact on the jet direction.

[0095] It should be understood that aspects of the non-rotating annular collector can be configured in a similar manner to the annular reservoir. However, those skilled in the art will understand that design differences may exist between the annular collector and the annular reservoir, since the annular collector is preferably independent of the rotation of the output unit, for example, non-rotatable, while the annular reservoir rotates with the helical channel / reactor bed or each helical channel / reactor bed.

[0096] The annular collector may or may not be part of the reactor. Preferably, the annular collector is independent of the rotation of the output unit, for example, it is non-rotatable. It should be understood that the inner surface of the annular collector is arranged to collect the jet. The annular collector has a curved upper section to collect the jet. The cross-section of the annular collector may be substantially U-shaped. The annular collector can ideally be configured to prevent liquid from escaping before it can be emptied from the collector.

[0097] The annular collector may include annular sidewalls arranged to capture the jet formed by the output unit. The annular collector prevents liquid from escaping the collector as it decelerates upon impact with the collector surface. The sides of the annular collector may be substantially shaped with a lateral U-shaped cross-section. Preferably, each side of the annular reservoir has a substantially J-shaped cross-section. Preferably, the sidewalls are integral with the base.

[0098] The annular collector may include a top (e.g., a lid) to help prevent liquid from leaving the annular collector once it has entered and / or to prevent contaminants from entering the annular collector. The annular collector (e.g., the top) may have an orifice (e.g., annular or circular orifice) to allow liquid to enter as the annular collector rotates. Preferably, the top is integral with the sidewalls of the annular collector.

[0099] Once the jet is captured by the collector, the liquid is radially decelerated and falls into the annular collector under gravity. Flow under gravity can be aided by using a negative gradient (i.e., a slope from a high point to a low point). The annular collector may include (e.g., at a low point or fluid-connected to a low point) one or more discharge orifices and / or reservoirs, or fluid-connected to a reservoir. The annular collector (e.g., a U-shaped section) may be axisymmetric about the (intended) axis of rotation, such that the flow along the collector is driven by the liquid height difference toward the discharge orifice and / or reservoir. Therefore, the annular collector can collect liquid that has passed through the reactor and store or transfer it to another component of the overall system.

[0100] Auxiliary components The reactor can be equipped with a motor to rotate the reactor bed. The reactor can also be equipped with a controller, such as a motor controller, to control the rotational speed.

[0101] The reactor may be equipped with devices for monitoring the reaction, such as detectors like cameras, NIR detectors, and / or diode array detectors. The reactor may be equipped with lighting equipment (e.g., LED lighting) to illuminate the helical channel through a at least partially transparent covering. Preferably, the reactor has a camera configured such that the frame rate is synchronized with the reactor's rotation frequency. This allows observation of a fixed area of ​​the helical channel, enabling photographs to be recorded for biofilm thickness measurement and / or continuous observation.

[0102] The reactor may include an airtight housing configured to meter the gas entering and exiting the helical channel.

[0103] The reactor may include a manifold to switch between different liquid input streams.

[0104] The reactor may include means for controlling the temperature of the reaction conditions within the helical channel. This may be means for raising the temperature of the helical channel, such as a heater (e.g., a thermoelectric heater) and / or means for lowering the temperature, such as an ice bath or thermoelectric cooler. The means for controlling the temperature may include conduits for a temperature-controlled fluid (e.g., water), wherein the temperature-controlled fluid may be heated or cooled by an external device. Preferably, the temperature within the helical channel is 0 to 100°C, such as 1 to 80°C, or 5 to 50°C, preferably 20 to 50°C, or 20 to 45°C, more preferably 25 to 40°C, such as 30 to 40°C, or 35 to 40°C. As those skilled in the art will understand, the selected temperature may be chosen for the reaction that is suitable to occur. The reactor may include insulation to maintain the temperature within the helical channel. Heat transfer and / or subsequent temperature uniformity throughout the reactor may be aided by any gas flow generated and driven by the rotating reactor unit within the reactor housing.

[0105] The reactor may include a rotatable shaft configured to rotate together (i.e., in unison) with the reactor bed, receiver unit, and output unit, or each of the reactor bed, receiver unit, and output unit. Alternatively, the reactor bed, receiver unit, and output unit, or each of the reactor bed, receiver unit, and output unit, may be configured to rotate together about a static shaft.

[0106] The reactor may include a gas inlet and / or a gas outlet, for example, having corresponding gas inlet conduits and / or gas outlet conduits. The gas inlet and / or gas outlet may be fluidly connected to a helical channel. The gas inlet conduit and / or gas outlet conduit may be coaxial with the intended axis of rotation. The gas inlet conduit and gas outlet conduit may be coaxial with each other. The gas inlet conduit and / or gas outlet conduit may be at least partially defined by a shaft. The gas inlet (conduit) may be fluidly connected to the outer end (e.g., outer common connection) of the helical channel or each helical channel, and / or the gas outlet (conduit) may be fluidly connected to the inner end (e.g., inner common connection) of the helical channel or each helical channel. The gas outlet (conduit) may additionally or optionally be fluidly connected to the inner end of a device (e.g., a weir) for controlling the pressure in the helical channel or each helical channel.

[0107] The amount and pressure of gas fed into the spiral channel can be controlled by devices such as regulators and / or valves. When introducing and / or collecting a gas phase from the spiral channel, sealed connections are typically used at the inlet and / or outlet to create the required pressure differential and / or ensure that the outlet gas can be collected independently.

[0108] Technicians will understand that various structures can be used to support the reactor and its rotating components. The reactor may be supported at both ends or may be cantilevered. The reactor may include a shell, for example, for housing and / or supporting the reactor bed. The shell may include one or more bearings and flanges supporting the reactor bed. When the bearings and / or flanges are located at both ends of the reactor bed (e.g., above and below), multiple supports may space the flanges. The structure of the reactor shell can ideally be designed to avoid mechanical resonance within the operating range of rotational speeds.

[0109] In use, the reactor can be positioned and / or configured such that the helical channel is substantially horizontal and the axis of rotation of the reactor is substantially vertical. Preferably, the axis of rotation is substantially perpendicular to the plane of the helical channel.

[0110] The reactor components can be formed from any suitable material. This material can be a metal, such as stainless steel, aluminum and / or chromium, resin, plexiglass, glass, or a mixture thereof. The material can be a metal selected from the list of metals composed of stainless steel, aluminum, and titanium and their alloys. The material can be a resin, plastic (e.g., thermoplastic and / or acrylic resin), glass (e.g., borosilicate glass), or a composite thereof. Composite materials can be glass fiber and / or carbon fiber composites.

[0111] Suitable thermoplastics include polyaryletherketone (PAEK), polyetherketoneketone (PEKK), and polyetheretherketone (PEEK). Other suitable plastic materials include polyetherimide (PEI), polysulfone (PPSU, PSU, PES), and polyphenylene sulfide (PPS). PEEK is preferred due to its high chemical resistance and structural stability.

[0112] Final substrate and precursor material The final matrix of the method of the present invention is selected from esters, fatty acid esters, acids, alcohols, aldehydes, apocarotenoids, and sesquiterpenes. Preferably, the final matrix of the method of the present invention is selected from ethyl acetate, isoamyl acetate, isoamyl alcohol, benzaldehyde, 2-phenylethyl acetate, 2-phenylethylbutyrate, phenylethylpropionate, methyl jasmonic acid, 2-phenylethanol, C2-C6 carboxylic acids, hexanal, octanal, decanal, dodecaldehyde, vanillin, apocarotenoids (preferably α- and β-ionones), and sesquiterpenes. More preferably, the final matrix is ​​selected from ethyl acetate, isoamyl acetate, isoamyl alcohol, benzaldehyde, and 2-phenylacetic acid. Ethyl esters, 2-phenylethyl butyrate, phenylethyl propionate, methyl jasmonic acid, 2-phenylethanol, C2-C6 carboxylic acids, hexanal, octanal, decanal, dodecaldehyde, vanillin, α- and β-ionones and sesquiterpenes, and even more preferably the final matrix is ​​selected from ethyl acetate, isoamyl acetate, isoamyl alcohol, benzaldehyde, 2-phenylethyl acetate, 2-phenylethyl butyrate, phenylethyl propionate, methyl jasmonic acid, 2-phenylethanol, and most preferably 2-phenylethanol.

[0113] Suitable and preferred yeast strains for producing the final substrate described above include *Clavulatus lucida*, *Kluyveromyces marsupialis*, species of *Kluyveromyces*, *Kluyveromyces dobii*, *Saccharomyces cerevisiae*, *Clavulatus delbrueckii*, non-yeast yeasts from Durango agave, *Saccharomyces cerevisiae*, *Schizosaccharomyces cerevisiae*, *Rhodotorula spp.*, species of *Pyrrosia*, *Beauveria bassiana*, *Porphyra yezoensis*, *Aspergillus niger*, and *Yersinia lipolytica*.

[0114] Suitable and preferred bacteria for producing the aforementioned final matrix are selected from Escherichia coli, Streptomyces, Pseudomonas, Acetobacter, Staphylococcus, and Lactococcus. These can be particularly useful materials for producing vanillin, apocarotenoids, and C2-C6 carboxylic acids.

[0115] Suitable and preferred enzymes for producing the final matrix described above are selected from alcohol dehydrogenases (ADHs) (e.g., those derived from horse ( E. Caballus ) or thermophilic denitrifying Bacillus ( G. Thermodenitrificans ), flavin-dependent alcohol oxidases (e.g., from S. Chinensis ), copper free radical oxidase (e.g., from Anthracnose bacterium granatum ( C. Graminicola ), lipases, and acetyltransferases. These may be particularly useful for the production of hexanal, octanal, decanal, dodecaldehyde, and other materials such as menthol and cinnamyl alcohol.

[0116] Preferred sesquiterpenes that can be produced by the method of the present invention include α-farnescene, β-farnescene, trans-nerolidol, α-santalol, patchulol, longifolene, α-humulene, elemol, cedrol, cubenol, guaiacol, α-linalofuran, β-linalofuran, aristolochine, linaloene, valenene, and noookatone. These can be suitably produced using brewer's yeast (α-farnescene to linaloene) and lipolytic yeast (valenene and noookatone). The final matrix sesquiterpenes α-farnescene and β-farnescene can be suitably produced by brewer's yeast or lipolytic yeast.

[0117] The precursor material used depends on the desired final matrix. The precursor material can be suitably selected from materials that, under biotransformation, particularly by yeast, bacteria, or enzyme biocatalysts, produce the desired final matrix. These biotransformations are known in the art.

[0118] Examples of suitable precursor materials include L-phenylalanine for the production of 2-phenylethanol, glucose, maltose, or maltotriose for the production of esters, glucose or farnesyl diphosphate for the production of sesquiterpenes, glucose, glycerol, xylose, eugenol, or ferulic acid for the production of vanillin, pyruvate for the production of C2-C6 carboxylic acids from glucose, hexan-1-ol for the production of hexanal, octan-1-ol for the production of octanal, decan-1-ol for the production of decanal, and dodecan-1-ol for the production of dodecaldehyde.

[0119] To produce apocarotenoids, such as α- and β-ionones, precursor materials include β-carotene. Typical methods for preparing culture media and fed-batch media can be found in Zhang et al., Biotechnology and Bioengineering. 2018;115:174-183.

[0120] The precursor material is a fluid (e.g., a liquid or a gas), preferably a liquid.

[0121] Precursor materials can typically be added to a spiral bioreactor as part of the growth medium composition, which may also contain materials that contribute to the conversion reaction.

[0122] The growth medium is typically specific to the yeast and the reaction to be transformed, a fact readily available in the art. The reactions and media are known, and the application of these known reaction pathways in a helical bioreactor offers advantages over the claimed method compared to conventional continuous stirred tank reactors.

[0123] An exemplary growth medium composition for converting L-phenylalanine (L-PA) to 2-phenylethanol (2-PE) using the yeast Kluyveromyces martensii (CBS 600) is: Glucose, L-phenylalanine, magnesium sulfate, citric acid, dipotassium hydrogen phosphate, yeast nitrogenous base without amino acids and ammonium sulfate, sodium acetate buffer.

[0124] Liquid growth media containing precursor materials can be suitably pretreated with a gas selected from air, oxygen, and / or carbon dioxide. Such pretreatment may be advantageous in preparing growth media for reaction with biocatalysts.

[0125] Biofilm coating of the spiral channel For the term biofilm, we define it as a biological entity. In this invention, yeast, mold, fungus, bacteria, or enzymes are catalytic materials present in the biofilm. Preferably, the catalytic material present in the biofilm is yeast, bacteria, or enzymes, more preferably yeast.

[0126] The coating of helical channels with biofilms is preferably carried out in the following manner: a) i) Loading a suspension containing particles of catalyst material into a spiral channel; and ii) While the suspension is inside the spiral channel, rotate the spiral channel for a period of time; iii) Optionally, but preferably, the supernatant is removed; or b) i) While the helical channel rotates, a suspension containing particles of catalyst material is loaded into the helical channel; and ii) Continue the rotation of the spiral channel for a period of time; iii) Optionally, but preferably, the supernatant is removed.

[0127] The biofilm coating process may include removing the supernatant (i.e., the liquid obtained after particle deposition).

[0128] Preferably, the coating of the helical channel with a biofilm is carried out via b) above, that is, while the helical channel is rotating, a suspension containing particles of catalyst material is loaded into the helical channel. Loading during the rotation of the helical channel helps to prevent the fixed particles on the outer wall from becoming loose under gravity and to be discharged between the injection of the suspension and the rotation to a certain speed.

[0129] The average particle size can be 10 nanometers or larger, such as 100 nanometers or larger, or 500 nanometers or larger, such as 800 nanometers or larger. The average particle size can be 100 micrometers or smaller, such as 50 micrometers or smaller, or 20 micrometers or smaller, such as 10 micrometers or smaller, such as 1 micrometer or smaller. The average particle size can range from 10 nanometers to 100 micrometers, such as from 500 nanometers to 50 micrometers. The average particle size can be determined, for example, by laser diffraction. The average value can be a number, volume, surface area, or other average value (preferably a number average value).

[0130] The rotational speed can be 100 RPM or higher, such as 200 RPM or higher, or 300 RPM or higher, preferably 400 RPM or higher, such as 600 RPM or higher, such as 1000 RPM or higher, or 1500 RPM or higher. The rotational speed can be 100,000 RPM or lower, such as 50,000 RPM or lower, or 20,000 RPM or lower, or 10,000 RPM or lower, such as 8000 RPM or lower, or 4000 RPM or lower, or 3000 RPM or lower. The rotational speed can be from 100 RPM to 100,000 RPM, such as 400 RPM to 100,000 RPM, or 400 RPM to 20,000 RPM, or 1000 RPM to 10,000 RPM.

[0131] Those skilled in the art will understand that during suspension injection, high flow rates (and therefore Reynolds numbers) are ideally used to achieve sufficient agitation, and high rotational speeds are the most efficient way to achieve high flow rates. Various operations, such as cleaning spirals, benefit from high flow rates.

[0132] As extraction or gas generation occurs, the phase meniscus decreases at high rotational speeds. This causes the sliding gas pocket to press against the inner wall, without obstructing the particle bed, and makes the extraction layer more uniform, thus placing the phase interface at an equidistant distance from the particle bed in all locations.

[0133] The method may include filling the helical channel with a suspension through the inner end of the helical channel. The method may also include sealing the outer end of the helical channel before the suspension is loaded—this is possible because rotation allows gas initially in the helical channel to pass through the incoming suspension, thus preventing gurgling and inward flow restriction. The positioning of the outlet weir required for reactor operation ensures that the suspension will be contained, provided that the volume of the injected suspension matches the volume of the helical channel leading to the outlet weir and the outlet passage. When multiple helical channels are in operation, the suspension volume can ideally be substantially matched to the combined volume to fill all channels. Preferably, the volume of suspension loaded into the reactor is approximately the same as the volume of the reactor's helical channels (e.g., differing from the actual volume by ±10%, ±5%, or ±2%).

[0134] This period allows the particles to become deposits on the outer wall of the spiral channel. The method may include keeping the outer end of the spiral channel closed for a period of time during which particles are allowed to settle under rotation. This period may be 1 second or longer, such as 5 seconds or longer, or 10 seconds or longer, such as 15 seconds or longer, or 20 seconds or longer, such as 30 seconds or longer. The period may also be 24 hours or less, such as 10 hours or less, or 1 hour or less, such as 10 minutes or less. Furthermore, the period may range from 1 second to 24 hours, such as 10 seconds to 10 hours, or 15 seconds to 1 hour.

[0135] More specifically, a biofilm is coated onto the helical channel. Alternatively, the biofilm can be considered a bed of cells or, alternatively, a particle bed containing a biocatalyst. An example using yeast as a catalytic material is that yeast is a biocatalyst for the reaction that converts precursor materials into the final matrix. Before the introduction of the precursor material, yeast is introduced and coated onto the helical channel. The yeast material is thus contained within a biofilm located on the helical channel. The helical channel is pre-coated with a yeast-containing biofilm before the precursor material is added. This can be done some time before the addition of the precursor material, or preferably as a first step of the method. For example, a preferred first step is to coat the helical channel with biofilm material (also known as particle bed material), preferably while the helical channel is rotating (at a speed of 200 RPM or higher, such as 300 RPM or higher, or 400 RPM or higher), preferably above 400 RPM, while the volume of the material ideally matches the volume of the available helical channel. Rotation allows the yeast material to flow downwards and backfill the helical channel, settling into the coated biofilm on the helical channel.

[0136] Biofilm coating (so-called bed formation) is a separate process from the operation of the helical bioreactor that produces the final substrate, and it has its own characteristics. Coating can be performed by injecting a suspension and using a flow rate and rotation speed that provides sufficient Coriolis secondary flow motion to keep cells suspended at the cell inflow location. Thus, the bed is formed by the sedimentation of a homogeneous suspension under centrifugal acceleration, resulting in a uniform bed on the outer wall and along the length of the helical channel. The bed can grow from a very thin initial bed to the desired bed thickness. If cell growth continues, the desired thickness can be maintained as cells are removed when a critical fluid shear stress (critical shielding number) is reached.

[0137] In a preferred embodiment, the biofilm material is injected as a suspension at a sufficiently high flow rate such that the secondary motion associated with Coriolis acceleration is strong enough to counteract the particle settling rate. Particularly suitable conditions are a flow rate of 50 mL / min and a rotation speed of 1200 rpm. Once injected, the particles, in the case of yeast, settle onto the wall within seconds to provide a helical channel for biofilm coating. This provides the advantage of a particularly uniform biofilm coating along the length of the helical channel. The time required for sediment (biofilm coating) formation depends on the particle size, shape, and density in the given liquid medium.

[0138] The biofilm coating (so-called bed volume) can be increased by using a thicker cell bed, thus increasing reactor yield. Reactor yield can also be increased by stacking multiple helical channels from a common fluid inlet to parallel operation of the reactor, and by using longer helical channels (i.e., each helical channel rotates more times).

[0139] Biofilm coatings for helical channels can provide coated helical channels with a granular layer on the outer wall of the helical channel. This layer can be 10 nanometers thick or more, such as 1 micrometer thick or more, 100 micrometers thick or more, or 200 micrometers thick or more. The thickness of this layer can be 5 millimeters or less, such as 1 millimeter or less, 500 micrometers or less, or 400 micrometers or less, such as 300 micrometers or less. The thickness of this layer can range from 10 nanometers to 1 millimeter, or from 1 micrometer to 500 micrometers. Biofilm-coated helical channels may or may not contain liquid.

[0140] In a similar manner, other catalyst materials can be coated onto the helical channels.

[0141] The preferred method for introducing enzymes is to immobilize them on high surface area solids such as silica, alumina, or clay. The preferred particle size for such immobilized enzymes is in the range of 5-50 micrometers.

[0142] Reaction conditions It should be understood that reactors can be used for reactions involving two or three or more different stages. For example, a rotary helical reactor can facilitate reactions and / or mixing between two different gaseous phases; a gaseous and a liquid phase; a gaseous and a solid phase; two different liquid phases; and / or a liquid and a solid phase. Reactors can be used for three-phase systems, such as liquid-liquid-solid systems and / or gas-liquid-solid systems, or for four-phase systems: liquid-liquid-gas-solid. The solid phase can substantially coat the entire inner surface of the helical channel and / or can coat only a portion of the inner surface of the helical channel (e.g., the outermost wall of the helical channel).

[0143] The reactor can be loaded (e.g., pre-loaded) with a solid phase, thereby forming a layer on one or more walls of the helical channel. Preferably, any solid phase is present on the outer wall of the helical channel. Solid reactants can be loaded onto the reactor by injecting a suspension into the receiver unit. Such a suspension can be injected into the receiver unit, for example, using a syringe (e.g., driven by an injection pump).

[0144] The bioreactor rotation speed and temperature, reaction time, and any other reaction conditions used in the method of this invention can be adjusted around the conditions already described above.

[0145] DETAILED DESCRIPTION OF THE DRAWINGS Figure 1 The results (dashed line) show the increased productivity of 2-phenylethanol (2-PE) per reactor volume achieved using the spiral bioreactor (circle symbol) used in this invention at a stable temperature of 33°C, compared to the continuous stirred tank reactor (CSTR) with and without Gao and Dauglis (2009) at 35°C. Figure 1 The results showed that there was no product removal in the CSTR (which is typical in current industrial practice), compared to a production rate of 0.05 g / L / h (lower than...). Figure 1 (dashed line in the text), and CSRT-ISPR with product removal (more than 0.43 g / L / h production rate (higher than Figure 1 (The dotted line in the image). All results were obtained using the same yeast strain, Kluyveromyces martensii (CBS 600). The x-axis represents time in minutes, and the y-axis represents the reactor's productivity per unit volume in grams per liter per hour (g / L / h).

[0146] Figure 2A and 2B A photograph of the spiral channel is shown, revealing a 280-micrometer-thick Max Kluyveromyces biofilm on the outer wall, which was formed by sedimentation under centrifugal acceleration of 50 g. Figure 2B yesFigure 2A An enlarged and annotated version of a portion. Figure 11 The biofilm surface can be clearly seen in b (20). The biofilm is 280 micrometers thick and extends over a spiral channel with a length of 79 cm and a depth of 4 mm; the spiral channel is 1.5 mm wide.

[0147] The attached diagram Figure 3 A reactor 30 for use in this invention is shown. The reactor 30 has a reactor bed 32; a motor 34 for driving the rotation of the reactor bed; a flow valve 36 for controlling the liquid inlet; a product outlet 38 for sampling; a motor controller 300 for controlling the rotational speed; a thermocouple 302 inserted into a collector through a bottom flange of the housing to determine the reactor temperature; and a camera 304 for monitoring the reaction under illumination by a lamp 306. The motor controller 300 allows the camera 304 and the lamp 306 to be triggered synchronously with the rotation of the reactor bed 32.

[0148] Figure 3 The main features of the rotary screw reactor are shown. Electronic components (motor 34, motor controller 300, camera 304, and LED light 306) are located at the top along with the liquid inlet and outlet. Supports are also present at the top for mounting valves required to switch inlets between the culture medium flow, inoculum (suspension) injection, and air (part of flow valve 36). Three cylindrical pillars 308 separate the top flange 3000a and the bottom flange 3000b, housing bearings and seals (not shown) to protect the bearings from liquid and prevent leakage from the shell during sterilization of the device using liquid solutions. Figure 3 The rotating spiral reactor is shown, with no fixed sides, so the rotating units can be seen.

[0149] The attached diagram Figure 4A and 4B The reactor 40 used in the method of the present invention is shown. Figure 4A The reactor 40 displays a motor 42; a controller 44; a camera 46; a light-emitting diode (LED) lamp 48; a selector valve 400; a housing priming / exhaust port 402; a compressed air inlet 404; a liquid feed bottle 406; a dual injection pump 408; a check valve network 4000; a back pressure valve 4002; an inlet heater 4004; a reactor outlet 4006a; a housing discharge 4006b; a housing overflow 4006c; a wastewater discharge 4006d; and an inoculation pump 4008.

[0150] Figure 4A and 4BA broader arrangement is shown, including auxiliary equipment for reactor 40, including an injection pump 4004 for injecting inoculum, a dual injection pump 408 combined with a check valve network 400 to allow for a continuous flow of culture medium over extended periods, and a compressed air connection 404 to apply pressure for rapid discharge of clean liquid and to provide priming air for oxidizing the liquid. Reactor 40 also includes a camera 46 to allow photographic visualization of the biofilm material deposition within the helical channel. It should be understood that the dual injection pump 408 can be replaced by a continuous pump, and heat exchangers (e.g., to the top and bottom flanges) can be added to control the reactor temperature (e.g., the temperature within the helical channel).

[0151] The attached diagram Figure 5 A cross-section of the reactor 50 used in the method of the present invention is shown, which reveals as follows: Figure 3 The internal operation of reactors 30 and 40 is shown in Figures 4 and 5. Reactor 50 has a liquid inlet pipe 52; a feed reservoir (annular reservoir of the receiver unit) 54; an internal connection 56; a spiral channel 58 in the reactor bed 580; an external connection 500; a weir 502; an open channel 504; an overflow channel 506; and a collector / discharge pipe 508. Fluid flow path A is shown as passing through reactor 50 from liquid inlet pipe 52 to overflow channel 506. Reactor 50 is supported at both ends of a rotatable shaft.

[0152] The attached diagram Figure 6 With similar Figure 5 The cross-section of the reactor 60 used in this invention is shown in the diagram. The reactor 60 has six reactor beds 62. Each reactor bed 62 has a helical channel 64. The helical channels 64 are connected in parallel to each other. Figure 6 A motor 66 for rotating the reactor bed is also shown.

[0153] Reactor 60 includes extraction flow capacity in a stack of 6 contactors. It can be used with particles and react simultaneously with extraction.

[0154] Figure 6The inlet pipe 68 (one for each liquid phase) into reservoir 600 and the connection pathway from reservoir 600 and from the lowest spiral channel 62 at either end of the spiral channel to the outlet of the corresponding phase are shown. Five other spiral channels operate in the same manner by connecting pathways spaced at 60-degree angles around the axis of rotation of the reactor. The overflow channel forming the liquid flow into the receiver is not shown. Reactor 60 includes an annular receiver scoop 602 (one for each liquid phase) that feeds into discharge channels 604 (one for each liquid phase, only one shown). Discharge channels 604 extend downwards from a high point to a discharge outlet, extending 180 degrees in both directions from the high point. Those skilled in the art will understand that the slope and width of discharge channels 604 can be optimized for desired flow rates. Reactor 60 is cantilevered (i.e., supported at one end of a rotatable axis).

[0155] like Figure 6 As shown, the flow from the six weirs at each phase outlet is drained into two overflow channels. Combining the flow from multiple weirs into the same overflow channel and / or increasing the flow rate allows the user to retain more angular momentum in the annular collector when removing particles, which may help remove debris from the collector.

[0156] The attached diagram Figure 7A -C shows a cross-sectional view of reactor 70, which is an alternative design for the reactor that can be used in this invention. Figure 7A It shows weir 72, and Figure 7B Overflow channel 74 and discharge port 76 are shown. For each liquid phase, reactor 70 includes two weirs 72, two overflow channels 74 and one discharge port 76. Figure 7C A plan view of the spiral channel 78 in the reactor bed 700 is shown. In general, Figure 7 further illustrates the fluid flow path through the reactor that can be used in this invention.

[0157] The attached diagram Figure 8 Is it like this? Figure 6 Photograph of reactor 70 shown in Figure 7.

[0158] Figure 9A -F shows a schematic cross-sectional view of different configurations of the reactor that can be used in this invention. Five helical channels are shown in each case, although only the fluid connection for one helical channel is shown. Due to the rotational displacement between the channels, the cross-sections of the helical channels appear staggered.

[0159] The attached diagram Figure 9A It showed something similar to Figure 6 The diagram shows a cross-sectional view of the reactor shown in Figure 7. Reactor 90 is configured to process a liquid phase. A particle bed (i.e., a solid phase) may be present on the outer wall of the channel.

[0160] In use, the helical coil is positioned substantially horizontally, and the axis of rotation is substantially vertical. For example... Figure 9A As shown, at the top, liquid (91) is fed through inlet pipe 92, causing the liquid to deposit on the platform 94a of the rotating feed reservoir 94b. Due to the rotation of the feed reservoir 94b, the liquid accelerates towards the outer edge of the feed reservoir 94b. This liquid merges with the reservoir liquid, filling to position "a" which depends on factors such as weir position and liquid feed rate.

[0161] Liquid flows downward through vertical conduit 96a and through helical channels 98 to their inner ends. Only one of the five helical channels 98 shown is depicted connected to vertical conduit 96a, but all helical channels (fluid) are connected to feed reservoir 94b. After moving through the helical channels 94b, the liquid flows downward along vertical connection 96b to radial channel 900, returning the liquid to the radial position of weir 902, and then vertically downward over weir 902. Each helical channel 98 shown is connected to a separate weir 902. Each weir 902 establishes a fixed hydrodynamic reference “d” (combining pressure, gravity, and centrifugal energy), over which the reservoir level at the top (a) is forced to exceed the reference depending on the amount of frictional loss in the helical flow to establish the flow of liquid through the helical channels. The choice to provide a weir for each helical channel, rather than for multiple or all helical channels, prioritizes design simplicity, as the structure is simply repeated every 360 / n degrees for n helical channels, with the helical channels arranged on different reactor beds in a stack. Furthermore, the identical flow path of each helical channel in the stack obtained by this method promotes a more even distribution of flow to the helical channels and can better adapt to structural requirements.

[0162] Once past the weir, the liquid flow enters directly into the overflow channel 902 (although the annular open channel can be used to deliver liquid to the overflow channel 902), causing the liquid to form a liquid jet. The jet is captured by the cup-shaped wall of the fixed annular collector 904. The bottom of the annular collector slopes downwards in two directions from the highest point to the discharge outlet, which is the reactor outlet and leads to the sampling device (e.g., Figure 7B As shown and combined Figure 7B (Describe it).

[0163] Figure 9A The flowchart depicts the fluid network of the rotating helical reactor. Liquid is introduced into the system through liquid inlet pipe 92, which fills the feed reservoir 94b. The liquid travels down along the inner connection to enter the helical channel at the inner end "b" and exit the helical channel at the outer end "c", and then proceeds to the weir 902 through the outer connection.

[0164] A gas (99) outlet (vent) is included at the inner end of the spiral "g" to allow displaced gas to leave during the injection of the particulate suspension to form a bed, and to allow any accumulated gas pockets to escape. The vent also allows any trapped or generated gas (99) to escape without disrupting the liquid inlet flow. In cases where the reaction depends on gaseous substances absorbed into the liquid, this can be achieved by saturating the receiver unit with gas. This is positioned more centrally than the inner end of the spiral channel, weir 902, and reservoir level "a" to avoid liquid loss through the vent.

[0165] When fluid is not being transferred into / out of the helical channel using a receiver / output unit, an injection pump (e.g., a dual injection pump, such as one combined with a check valve network) and / or a gas regulator can be used, such as... Figure 1 The system shown in Figure 2 is used to control the fluid input.

[0166] Rotating helical channels allow biofilms to be stabilized by centrifugation, by maintaining a continuous supply of media, and by potentially regulating product concentration through parallel-flow solvent extraction.

[0167] The reactor used in the method of the present invention can be configured to process two fluid phases (e.g., of different densities). The acceleration caused by the rotation of the helical channel allows the denser phase to stabilize on the outer side of the helical channel. In other words, the denser phase is stabilized by centrifugal acceleration. The denser phase can be continuously supplied with a second, less dense phase (e.g., through a reaction process). The concentration of the product in the denser phase can be adjusted or controlled by extraction with a parallel or countercurrent solvent flow (e.g., in the case where the solvent has a lower density).

[0168] The attached diagram Figure 9B It showed something similar to Figure 9A The diagram shows a cross-sectional schematic of a reactor configured to process a single-phase liquid medium (9000) stream with a gas (9001) output. For example, the gas may be generated by a reaction occurring within the reactor.

[0169] Figure 9B An axial conduit in the shaft is shown to allow gas (9001) to flow out uncontaminated, and the gas can be collected. The gas slides upward along the spiral channel in the "bag," but is also partially absorbed into the medium, which results in gas formation in the radial medium outlet channel as the pressure decreases along the channel.

[0170] The attached diagram Figure 9C It showed something similar to Figure 9A The diagram shows a cross-sectional view of a reactor configured to process a single-phase liquid medium (9000) flow with a gas input (9002) and a gas output (9001).

[0171] Higher levels of supply gas than can be achieved by saturating the inlet medium at the platform require the gas to remain in contact with the medium along the length of the helical channel. This can be achieved by incorporating a gas inlet conduit (“bleed-in”) fluidly connected to the outer end of the helical channel. This can be achieved using a tube coaxial with the gas outlet orifice in the shaft, which typically requires a rotary seal.

[0172] The attached diagram Figure 9D It showed something similar to Figure 9A The diagram shows a cross-sectional view of a reactor configured to process two liquid phases (9003 and 9004) in countercurrent flow. This is precisely... Figure 6 And type 7 (although with 5 (instead of 6) spiral channels). This reactor can be used to extract components (e.g., solutes) from one phase into another. Extraction flows have been demonstrated in reactors based on this design, where bacteria or yeast are used as the solid phase. It should be understood that, depending on the interface location used (large or small interface location in the spiral channel), regulating reservoirs can be placed at either end of the spiral channel. The reactor has reservoirs at two input levels "a" and "e"; and weirs at two levels "d" and "f".

[0173] Figure 9D This illustrates the "breaking of gravitational links," whereby each phase is uniformly distributed to each helical channel in the reactor. The outlet pathway for the lighter (i.e., less dense) phase (9003) extends upwards from the common reference position "f" for each lighter phase weir to the top of the stack where the inlet for each heavier (i.e., more dense) phase (9004) ends. The column's dimensions allow for negligible frictional losses within the column of the lighter phase liquid it forms. This achieves two scenarios: First, the pressure at each heavy phase feed outlet is the same (equal to the hydrostatic pressure rise from the shell pressure at “f” to the pressure at the top of the light phase liquid column), and since the flow is also driven from the same reservoir reference through the same passage from “a”, the individual heavy feed flows are equal.

[0174] Secondly, as the flow moves vertically downwards along the stack from one channel to another, the pressure at points "b" and "c" increases due to the increase in hydrostatic pressure, affecting the flow velocity through each spiral channel. However, because the fluid has a uniform (light phase) density, the pressure difference between "b" and "c" remains constant, and this is what drives the flow.

[0175] The roles of the two phases can be reversed to achieve alternating interface positions within the helical channel segments (for liquid-liquid flow, there are always two distinct positions for the same flow velocities of the two phases). Thus, the static column can extend from the weir "d" to the top of the stack, where it contains the heavy liquid. The regulating interface at "g" is positioned in a similar manner at the inner end of the helix.

[0176] Figure 9D The unit version of the 6-channel design shown has been successfully operated across a range of extraction liquids in contact with water as the heavy phase, rotational speeds, and flow rates. The reactor has been successfully used for extraction liquids of butanol, octanol, isopropyl myristate, or oleic acid. The successful use of oleic acid, which has a viscosity 28 times higher than water, demonstrates that highly viscous liquids can be surprisingly tolerated by the reactor.

[0177] The attached diagram Figure 9E It shows the relationship with Figure 9D A cross-sectional schematic diagram of a similar reactor is shown, wherein the reactor is configured to process two liquid phases (9000 and 9005) and flow countercurrently with a gas output (9001) (e.g., by generation) (e.g., for extraction).

[0178] A gas escaping pathway (i.e., an exhaust port) can be added as before to allow for the generation of extracted gas.

[0179] The air bladder slides upward along the spiral channel as before, but now it will slide along with the extraction liquid (9005) layer. Therefore, there is a complex flow pattern in the two liquid phases.

[0180] The attached diagram Figure 9F It showed something similar to Figure 9A The diagram shows a cross-sectional view of a reactor configured to process two liquid phases (9000 and 9005) and flow countercurrently with the gas inlet and gas outlet (9001) (e.g., for extraction).

[0181] The reactor design is similar to Figure 9E As shown, but with gas feed (9002). This can be provided using gas evacuation at the outer end of the spiral channel. A tube coaxial with the gas outlet hole in the shaft is required, and this requires a rotary seal, as per [reference needed]. Figure 9C As stated above.

[0182] The attached diagram Figure 10A schematic cross-sectional view of a reactor with reactor beds A (101), B (102), and C (103) is shown, each reactor bed having multiple (five) helical channels. Each bed is fed to the next bed by adding some new reagent material as liquid reagents A (104), B (105), and C (106). Each reactor bed may contain a different solid phase (e.g., particle / cell type).

[0183] The attached diagram Figure 11 It is displayed as follows Figures 3-5 The image shows a bed of watermelon seed powder with a deionized (DI) water flow in a spiral channel of a reactor. Analysis of the image indicates that the bed depth uniformity is within 10%. Enzyme deposition in the watermelon seed powder is achievable.

[0184] Figure 12 This is a graph showing the initial thickness of the watermelon seed powder bed after suspension injection. Figure 12 The results show that the sedimentation of the suspension is proportional to the particle fraction.

[0185] The attached diagram Figure 13A The initial bed of Max Kluyveromyces cells was shown in, as Figures 3-5 Growth in the spiral channel of the reactor shown. The initial biofilm thickness was 120 micrometers.

[0186] The attached diagram Figure 13B Shown in such Figures 3-5 The growth of Max Kluwer yeast cells in the spiral channel of the reactor shown at the end of an 8-hour 2PE production run. The biofilm thickness was 200 micrometers after 8 hours.

[0187] The attached diagram Figure 14 Shown in such Figures 3-5 Oxygen is generated in the watermelon seed powder bed within the spiral channel of the reactor shown. Gas pockets can be observed on the inner wall of the spiral channel, and these pockets slide upwards along the channel when they reach a critical size. Gas is generated during continuous operation at 2400 rpm for 5 hours with a 7.4 mM H₂O₂ aqueous solution flowing at 1.1 mL / min. Watermelon seed powder contains catalase, which catalyzes the decomposition of hydrogen peroxide to produce water and oxygen.

[0188] EMBODIMENT Experimental methods and materials The reactor is used to prepare biofilm-coated spiral channels. An injection pump is used to drive the flow of the medium and extraction solvent. The liquid (medium) is saturated with oxygen to supply yeast. The pathways are sterilized using a 20 g / L sodium hypochlorite solution (10 min) followed by a 50 g / L sodium thiosulfate solution (10 min), and then filled with the liquid (medium). The initial biofilm is formed by the inoculum suspension injected along the medium flow path to fill the spiral channels and the pathways in and out of the spiral channels. A series of valves are then opened and closed to remove the inoculum remaining in the connecting pathways and replace it with the medium. Rotation is then initiated to force the cells in the suspension to settle onto the outer wall of the spiral channels, thus forming a sedimentation layer, i.e., the biofilm.

[0189] Example 1 Demonstration of improved performance compared to existing bioreactors As an example of the potential of the method of the present invention, the production rate of 2-phenylethanol was measured.

[0190] Yeast Kluyveromycin (CBS 600) Reaction The conversion of L-phenylalanine (L-PA) to 2-phenylethanol (2-PE) via the Ehrlich reaction pathway: C9H 11 NO2+ C5H6O5+ C 21 H 29 N7O 14 P2 to C8H 10 O + C5H9NO4 + CO2 + C 21 H 27 N7O 14 P2 Medium composition Glucose, 30 g / L L-Phenylanine, 7 g / L Magnesium sulfate, 0.5 g / L Citric acid, 10.3 g / L Dipotassium hydrogen phosphate, 27.1 g / L A yeast-based nitrogen source that is free of amino acids and ammonium sulfate, 6.8 g / L. Sodium acetate buffer, 100 mmol, pH 5.0 Experimental details Suspension: 0.03% cell volume fraction (resulting in an initial biofilm thickness of approximately 100 micrometers) Injection: 13 ml of cell suspension at 50 ml / min and 1200 rpm. Operating conditions: Media flow rate of 4 ml / min, 2000 rpm, reactor temperature of 33°C (steady state). Sampling: Collect 2 mL of sample from the reactor outlet every 15 minutes (HPLC determination of 2PE concentration). Running time: 5.5 to 8.5 hours Interpretation of results The performance of a bioreactor is measured by its productivity per unit volume (referred to here as "specific production rate"). Figure 1 The results show typical results of a curve plotted for this parameter relative to the time after reactor startup. The reactor is not preheated, and therefore most of the initial increase in production rate is due to the temperature rise.

[0191] By approximately 120 minutes, the reactor reached a stable temperature of 33°C for this run, and the continued increase in specific production rate was due to biofilm growth (the membrane thickness increased from approximately 100 μm post-inoculation to approximately 180 μm at the end of the 5.5-hour experiment). At the end of this period, the 2-PE specific production rate had reached 0.77 g / L / h. Gao and Daugulis reported the productivity results of CSTR (continuous stirred tank reactor) using the same yeast reaction system (Kluyveromyces martensii) with and without ISPR (in-situ product removal). Without product removal (which is typical in current industrial practice), the specific production rate was 0.05 g / L / h, while with product removal it was 0.43 g / L / h. Therefore, the results from the helical bioreactor are already twice that obtained with CSTR using ISPR and approximately 15 times higher than typical industrial practice.

[0192] The results from these stirred reactors are that the reagent concentrations are approximately four times higher than those used in the prototype, and if this higher concentration is used, the specific production rate of the prototype is expected to increase by four times. Therefore, the helical bioreactor used in the method of the present invention is expected to achieve a specific production rate nearly an order of magnitude higher than that of conventional reactors (even with product removal).

[0193] By selecting suitable precursor materials and yeast materials (which act as catalysts for converting precursor materials into final matrix materials), this successful demonstration of increased productivity of 2-phenylethanol materials can be extended to the production of other final matrix materials of this invention.

[0194] The product is readily obtained from the helical bioreactor through the liquid outlet. This arrangement allows for sampling of the effluent to observe the amount of product relative to the reactants, and the flow rate can be adjusted accordingly. In different but preferred embodiments, the helical bioreactor is arranged with countercurrent (parallel) flow of the extractant liquid to increase the yield by removing product material. Examples of such extractant liquids used for countercurrent flow are isopropyl myristate, oleic acid, and decane.

[0195] Rotating helical channels allow biofilms to be stabilized by centrifugation, supported by a continuous supply of media, and potentially modulated by extraction with parallel-flowing solvents.

[0196] Example 2 The attached diagram Figure 11 It shows in such Figures 3-5 The image shows a bed of watermelon seed powder with a deionized (DI) water flow in a spiral channel of a reactor. Analysis of the image indicates that the bed depth uniformity is within 10%. Enzyme deposition in the watermelon seed powder is achievable.

[0197] Figure 12 This is a graph showing the initial thickness of the watermelon seed powder bed after suspension injection. Figure 12 The results show that the sedimentation of the suspension is proportional to the particle fraction.

[0198] Example 3 The attached diagram Figure 13A The initial bed of Max Kluyveromyces cells was shown in, as Figures 3-5 Growth in the spiral channel of the reactor shown. The initial biofilm thickness was 120 micrometers.

[0199] The attached diagram Figure 13B Shown in such Figures 3-5 The growth of Max Kluwer yeast cells in the spiral channel of the reactor shown at the end of an 8-hour 2PE production run. The biofilm thickness was 200 micrometers after 8 hours.

[0200] Example 4 The attached diagram Figure 14 Shown in such Figures 3-5 EMBODIMENT Experimental methods and materials Demonstration of improved performance compared to existing bioreactors Yeast Reaction Medium composition Experimental details Interpretation of results Figure 1 Figure 11 Figures 3-5 Figure 12 Figure 12 Figure 13A Figures 3-5 Figure 13B Figures 3-5 Figure 14 Figures 3-5 The reactor shown depicts oxygen generation in a bed of watermelon seed powder within a spiral channel. Gas pockets are observed on the inner wall of the spiral channel, and these pockets slide upwards along the channel when they reach a critical size. Gas is generated during 5 hours of continuous operation at 2400 rpm with a 7.4 nM H₂O₂ aqueous solution flowing at 1.1 mL / min. Watermelon seed powder contains catalase, which catalyzes the decomposition of hydrogen peroxide to produce water and oxygen.

[0201] This example demonstrates the application of this method to enzymes used as catalyst materials.

Claims

1. A method for producing a final matrix selected from esters, fatty acid esters, acids, alcohols, aldehydes, apocarotenoids, and sesquiterpenes; In this process, a suitable precursor fluid material is added to a rotating spiral bioreactor with spiral channels coated with a biofilm. The biomembrane material includes a material that serves as a catalyst for converting the precursor material into the final matrix material; The catalyst material is selected from yeast, mold, fungus, bacteria or enzyme, preferably yeast, bacteria or enzyme, more preferably yeast; Its features The helical channel rotates for a period of time while the precursor fluid material is inside the biofilm-coated helical channel; The resulting final matrix is ​​then removed.

2. The method of claim 1, wherein coating the helical channel with the biofilm is performed in the following manner: a) i) Loading a suspension containing particles of the catalyst material into the helical channel; and ii) The spiral channel is rotated for a period of time while the suspension is inside the spiral channel; iii) Optionally, but preferably, the supernatant is removed; or b) i) While the spiral channel is rotating, a suspension containing particles of the catalyst material is loaded into the spiral channel; and ii) Continue the rotation of the spiral channel for a period of time; iii) Optionally, but preferably, the supernatant is removed.

3. The method according to claim 1 or claim 2, wherein the final matrix is ​​selected from ethyl acetate, isoamyl acetate, isoamyl alcohol, benzaldehyde, 2-phenylethyl acetate, 2-phenylethyl butyrate, phenylethyl propionate, methyl jasmonic acid, 2-phenylethanol, C2-C6 carboxylic acids, hexanal, octanal, decanal, dodecaldehyde, vanillin, apocarotenoids (preferably α- and β-ionones), and sesquiterpenes, preferably the final matrix is ​​selected from ethyl acetate, isoamyl acetate, isoamyl alcohol, ... Benzaldehyde, 2-phenylethyl acetate, 2-phenylethyl butyrate, phenylethyl propionate, methyl jasmonic acid, 2-phenylethanol, C2-C6 carboxylic acids, hexanal, octanal, decanal, dodecaldehyde, vanillin, α- and β-ionones, and sesquiterpenes, more preferably the final matrix is ​​selected from ethyl acetate, isoamyl acetate, isoamyl alcohol, benzaldehyde, 2-phenylethyl acetate, 2-phenylethyl butyrate, phenylethyl propionate, methyl jasmonic acid, 2-phenylethanol, and most preferably 2-phenylethanol.

4. The method according to any one of claims 1 to 3, wherein the yeast material is selected from *Clavulatus lucida*, *Kluyveromyces marsupialis*, species of *Kluyveromyces*, *Kluyveromyces dobii*, *Saccharomyces cerevisiae*, *Clavulatus delbrueckii*, non-yeast yeasts from Durango agave, *Saccharomyces cerevisiae*, *Schizosaccharomyces cerevisiae*, *Rhodotorula spp.*, species of *Pyrrosia*, *Beauveria bassiana*, *Porphyra yezoensis*, *Aspergillus niger*, and *Yersinia lipolytica*; wherein the bacteria are selected from *Escherichia coli*, *Streptomyces*, *Pseudomonas*, *Acetobacter*, *Staphylococcus*, and *Streptococcus lactis*; and wherein the enzyme is selected from alcohol dehydrogenase (ADH), flavin-dependent alcohol oxidase, copper radical oxidase, lipase, and acetyltransferase.

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