Chromatographic separation of three components of allulose from corn syrup

Abstract

The present invention is advantageous over conventional methods and products. In one embodiment, the method comprises separating a mixture of allulose, fructose, glucose, and glucooligosaccharides (wherein the separation comprises using simulated moving bed chromatography) and recovering allulose in high purity and yield. In one embodiment, simulated moving bed ("SMB") chromatography is used to separate the allulose-rich fraction from a fructose- and glucose-rich fraction, and to separate the fructose-rich fraction from a glucose-rich fraction. In one embodiment, the method provides for the separation of allulose from a mixture of allulose, fructose, and D-glucose, wherein the mixture is produced from high fructose corn syrup ("HFCS") when HFCS is contacted with allulose epimerase. In one embodiment, the method produces a high-quality allulose product. In one embodiment, the fructose-rich fraction can be recycled for contact with allulose epimerase.

Description

[Technical Field]

[0001] FIELD OF THE INVENTION The present invention relates to a process for producing allulose from corn syrup using simulated moving bed chromatography. [Background technology]

[0002] Background of the Invention

[0002] Allulose is a low-calorie sweetener. Conventionally, allulose is typically formed by isomerizing fructose to allulose using an immobilized epimerase enzyme. This process typically converts about 15-30% of fructose to allulose. Chromatographic fractionation has been used to separate allulose and fructose to produce high-purity allulose. Dissolved crystalline fructose is a typical feedstock for the enzymatic conversion of fructose to allulose. To produce crystalline fructose, a very high-purity fructose feedstock with a fructose purity greater than 96% is required.

[0003]

[0003] U.S. Patent No. 10,342,247 discloses the preparation of allulose from a crystalline fructose solution having a purity of approximately 99%. The patent discloses that crystalline fructose was dissolved in water, adjusted to a pH of 7.0, and the mixture was reacted with an epimerization enzyme at 55°C. After 40 hours of reaction, a syrup sample was collected having 25.2% allulose and 74.8% fructose as determined by standard HPLC. The patent discloses that the syrup was passed through microfiltration to remove the insoluble cell mass containing the enzyme, then subjected to carbon filtration to remove color, and then demineralized with cation and anion exchange columns to further remove minerals and other impurities. The patent discloses that the syrup was then concentrated using a conventional evaporator to approximately 60% dry solids. This patent discloses that the 25.2% concentrated allulose syrup produced in the above step was passed through a simulated moving bed chromatographic column (SMB) containing calcium-form resin, and the resulting syrup had an allulose content of 93%.

[0004]

[0004] A significant limitation to conventional methods is that crystalline fructose requires expensive and time-consuming processing for production, and crystalline fructose is a relatively high-cost feedstock. Summary of the Invention [Problem to be solved by the invention]

[0005]

[0005] There is a need for improved methods for producing allulose that do not have the drawbacks and limitations of conventional methods. [Means for solving the problem]

[0006] Brief Summary of the Invention

[0006] The present invention offers advantages over conventional methods and products. The present invention provides an efficient process that can convert readily available and lower-cost feedstocks, such as high fructose corn syrup ("HFCS"), to allulose. In an embodiment of the present invention, the method comprises separating a mixture of allulose, fructose, and glucose, and optionally gluco-oligosaccharides, wherein the separation comprises using simulated moving bed ("SMB") chromatography, and recovering allulose in high purity and yield. In one embodiment, the SMB chromatography disclosed herein is a ternary (also called "tertiary") chromatography that separates allulose from fructose and glucose in a first zone and simultaneously separates fructose from glucose in a second zone.

[0007]

[0007] In one aspect, a process for purifying allulose includes contacting a chromatographic bed material with water and a mixture of allulose, fructose (wherein fructose is amorphous fructose) and glucose, and simultaneously separating an allulose-rich fraction from a fructose- and glucose-rich fraction by adsorbent chromatography on the chromatographic bed material, and simultaneously separating a fructose-rich fraction from a glucose-rich fraction by adsorbent chromatography on the chromatographic bed material.

[0008]

[0008] In certain embodiments, a simulated moving bed apparatus including a plurality of column segments connected in series and containing chromatographic bed material comprises: a. feeding a mixture of fructose, dextrose, and allulose reaction products at a feed port location to contact the chromatographic bed material in the column segments of a first zone; b. feeding water into the apparatus at an eluent port location to contact the chromatographic bed material in the column segments of a second zone; c. flowing water in a first direction toward the first zone to contact the mixture of fructose, dextrose, and allulose reaction products and a first eluate enriched in allulose reaction product from a product port located in a second zone upstream of the feed port location in the first direction; e. a second eluate enriched in fructose from a first raffinate port location in the first zone downstream of the feed port in the first direction; and f. a third eluate enriched in dextrose from a second raffinate port location in a third zone downstream of the first raffinate port in the first direction.

[0009] In one aspect, the allulose production system comprises: a. a vessel containing immobilized allulose epimerase, wherein when high fructose corn syrup contacts the immobilized allulose enzyme, the allulose epimerase converts fructose to allulose; b. A simulated moving bed apparatus comprising a plurality of column segments connected in series and containing chromatographic bed material, the plurality of column segments being located downstream of a vessel containing immobilized allulose epimerase; c. a feed port for receiving a mixture of fructose, dextrose, and allulose from a vessel containing immobilized allulose epimerase to contact the chromatographic bed material in the column segment of the first zone; d. an eluent port in the column segment of the second zone for receiving water to contact the chromatographic bed material; e. the simulated moving bed is configured to flow water in a first direction toward the first zone to contact the mixture of fructose, dextrose, and allulose, and to move the plurality of column segments in a second direction opposite to the first direction; d. a product port disposed in a second zone upstream of the feed port location relative to the first direction for removing the allulose-rich first eluate; e. a first raffinate port for removing a fructose-rich second eluate, the first raffinate port being located in a first zone downstream of the feed port in the first direction; and f. a second raffinate port for removing a third dextrose-rich eluate, the second raffinate port being located in a third zone downstream of the first raffinate port with respect to the first direction; Includes:

[0010] In one aspect, the process involves contacting a chromatographic bed material comprising a resin with a mixture comprising fructose, dextrose (also known as D-glucose), and allulose reaction products. In one aspect, the mixture comprising fructose, dextrose, and allulose reaction products is produced by the enzymatic reaction of high fructose corn syrup with an immobilized epimerase to convert fructose in the syrup to allulose reaction products. The method includes separating an allulose reaction product-enriched fraction from a fructose- and glucose-enriched fraction, and separating the fructose-enriched fraction from a dextrose-enriched fraction. In one aspect, the fructose-enriched fraction has a larger volume than the dextrose-enriched fraction.

[0011] In one embodiment, the fructose-rich fraction can be recycled to the enzymatic reaction of high fructose corn syrup, so that at least a portion of the recycled fructose is converted to allulose. The advantage of recycling the enriched fructose fraction in this manner is that the enriched fructose fraction does not need to undergo evaporation before the enzymatic reaction to convert the fructose in the fraction to allulose.

[0012] In some embodiments, the dextrose-rich fraction can be recycled as a feedstock for an upstream fructose refining process in which dextrose is converted to fructose. In some embodiments, the upstream fructose refining process can be followed by an HFCS refining process. The HFCS refining process can produce an HFCS stream that can be used as a feedstock for the enzymatic reaction of high fructose corn syrup with an immobilized epimerase to convert the fructose in the syrup to an allulose reaction product.

[0013]

[0013] In one aspect, the chromatographic bed material is contained in a simulated moving bed apparatus. In a specific embodiment, the simulated moving bed apparatus comprises a plurality of movable column segments connected in a sequential, fluidly continuous manner, and comprises a desorbent port for introducing water into the apparatus to contact the chromatographic bed material, a feed port for introducing a mixture of fructose, dextrose, and allulose reaction products into the apparatus to contact the chromatographic bed material, a product port for removing high-purity allulose from the apparatus, a port for removing a fructose-rich fraction from the apparatus, and a port for removing a dextrose-rich fraction from the apparatus. Typically, the column segments are connected sequentially in a circular sequence. In this embodiment, water and a mixture containing fructose, dextrose, and allulose reaction products are each introduced into the simulated moving bed apparatus and passed through the apparatus in a first flow direction, and the column segments are collectively moved in a second direction opposite to the first direction.

[0014]

[0014] In another embodiment, the simulated moving bed apparatus includes a plurality of column segments fluidly interconnected by movable ports, wherein the movable ports are a desorbent port for introducing water into the apparatus to contact the chromatographic bed material, a feed port for introducing a mixture of fructose, dextrose, and allulose reaction products into the apparatus to contact the chromatographic bed material, a product port for removing high-purity allulose from the apparatus, a port for removing a fructose-rich fraction from the apparatus, and a port for removing a dextrose-rich fraction from the apparatus. In this embodiment, the movable ports are sequentially moved circularly relative to the column segments. Typically, water and a mixture containing fructose, dextrose, and allulose reaction products are each introduced into the simulated moving bed apparatus and pass through the apparatus in a first flow direction, and the movable ports are collectively moved in the same direction to adjacent column segments to simulate movement of the bed segments in a second direction opposite to the first direction.

[0015]

[0015] In one aspect, methods are provided for the continuous production of high-purity allulose by the simulated moving bed process described above. These methods operate by continuously contacting a chromatographic bed material with water and a mixture of fructose, dextrose, and allulose reaction products, along with the removal of a high-purity allulose-enriched eluate.

[0016]

[0016] According to an embodiment of the present invention, the chromatographic bed material is an ion exchange resin. Examples of suitable chromatographic bed materials include, but are not limited to, resins that are strong acid cation resins. Examples of suitable strong acid cation resins include those sold by Dow Chemical Co. (Midland, Michigan) under product numbers Dowex99 Ca / 320, Dowex99 Ca / 310, and Dowex99 Ca / 280; and those sold by Mitsubishi Chemical Co. (Tokyo, Japan) under the product name DIAION TM Examples include those sold at

[0017]

[0017] In yet another aspect, there is provided an allulose production facility including a simulated moving bed apparatus configured to operate any one of the processes provided herein. Such a facility may be characterized by having a simulated moving bed apparatus configured with ports that simultaneously direct the flow of water and trisaccharide feedstock to a chromatographic bed material, and by simultaneously removing the allulose product and fructose and dextrose in different zones of the simulated moving bed apparatus.

[0018]

[0018] These and other aspects, embodiments, and related advantages will become apparent from the following brief description of the drawings and detailed description.

[0019] BRIEF DESCRIPTION OF THE DRAWINGS

[0019] The features and advantages of the present disclosure may be better understood with reference to the accompanying drawings. [Brief explanation of the drawings]

[0020] [Figure 1] 1 is a schematic flow diagram of a simulated moving bed chromatography system having 12 columns in accordance with an embodiment of the present invention. [Figure 2]

[0021] 1 is a schematic flow diagram of a simulated moving bed chromatography system having 14 columns in accordance with an embodiment of the present invention. [Figure 3]

[0022] 1 is a schematic flow diagram of a simulated moving bed chromatography system having 20 columns in accordance with an embodiment of the present invention. [Figure 4]

[0023] 1 is a flow diagram illustrating the production of allulose from HFCS in accordance with an embodiment of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0021] Detailed Description of the Preferred Embodiments

[0024] Before describing the present invention in detail, in order to better distinguish between subtleties of meaning that may apply to different embodiments of the present invention, certain terms having meanings that are generally understood by those of ordinary skill in the art are nevertheless defined herein. The definitions provided herein are intended to encompass, without limitation, the ordinary meanings understood in the art, unless such meanings are in conflict with the definitions provided herein, in which case it is understood that the definitions provided shall control.

[0022]

[0025] "Chromatographic separation" and its variants refer to the velocity-based separation of chemical species on a stationary solid-phase chromatographic sorbent material by differentially partitioning the species between the stationary phase and the mobile eluent phase. "Velocity-based separation" means that a portion of each species always travels with the mobile phase eluent, but differences in partitioning between the species result in different migration rates of the species on the stationary phase, thereby achieving a separation that is time- and bed-volume-dependent. Thus, chromatographic separation can be achieved by using a single mobile phase without the need to change eluent conditions. In this regard, chromatographic separation can be characterized as a "continuous separation process" because the species being separated are in continuous motion.

[0023]

[0026] "Sorption" is the process by which a substance becomes physically bound to a substrate material by absorption or adsorption.

[0024]

[0027] "Absorption" is the process by which molecules or ions of a substance become physically bound within the substrate material by becoming entrapped within the matrix of the substrate material, i.e., the substrate material is an absorbent.

[0025]

[0028] "Adsorption" is the process by which molecules or ions of a substance attach to the surface of a substrate, thereby becoming physically bound to the substrate material, i.e., the substrate material is an adsorbent.

[0026]

[0029] "Adsorption / desorption separation" or "adsorption / desorption chromatography" and variations thereof are specifically meant to distinguish from chromatographic separation and refer to a process in which chemical species are separated by immobilizing one species on a solid-phase adsorbent while preferentially transporting another species with the eluent under first eluent conditions, and then changing the eluent conditions to second conditions in which the immobilized species desorbs from the adsorbent and preferentially partitions into the eluent. Thus, two features fundamentally distinguish adsorption / desorption separation from chromatographic separation: (i) first eluent conditions are selected to achieve as nearly complete immobilization as possible of at least one species on the chromatographic material, and (ii) there is a change to second eluent conditions selected to render the immobilized species mobile. In this regard, adsorption / desorption separation can be characterized as a "discontinuous" or "stepwise" separation process.

[0027]

[0030] Those skilled in the art will recognize that, depending on the choice of eluent and chemical species, the same solid-phase chromatographic material can be used to achieve chromatographic separation, adsorption / desorption separation, or both. For example, in a first step to separate a mixture containing species A, B, and C, an ion exchange material can be used as the adsorbent stationary phase with an eluent of a first pH selected to cause species A and B to immobilize on the adsorbent, while species C preferentially partitions into the mobile eluent phase, thereby achieving adsorption / desorption separation of species A and B from species C. In a second step, the eluent conditions can be changed to a second pH that causes species A and B to preferentially partition into the mobile phase. Furthermore, if the pH change causes species A and B to differentially partition between the mobile and stationary phases, species A and B will migrate at different rates on the stationary phase and be chromatographically separated on the same ion exchange material.

[0028]

[0031] "Adsorbent separation" or "adsorbent chromatography" and grammatical variations thereof generally refer to the separation of chemical species using a solid-phase adsorbent material and at least one mobile-phase eluent. These terms encompass both chromatographic separations and adsorption / desorption separations.

[0029]

[0032] "Chromatographic bed material," "chromatographic adsorbent," or "stationary phase" refers to a solid-phase adsorbent material used to separate chemical species by adsorbent separation.

[0030]

[0033] "Eluant" refers to the fluid mobile phase that is passed over a chromatographic bed material to achieve adsorbent separation.

[0031]

[0034] "Raffinate" is a general term that refers to the liquid eluate or fraction obtained from a separation procedure that is not enriched in the desired product allulose.

[0032]

[0035] "Continuously operated" or "continuously separated" in the context of the use of an adsorbent chromatographic separation process means that the process is carried out indefinitely with respect to time, with a continuous input of reactants and / or eluent, a continuous withdrawal of product and / or raffinate, and, optionally, a continuous flow of bed preparation material. In this regard, both adsorption / desorption separations and chromatographic separations can be operated continuously, the difference being that in adsorption / desorption separations, some sections of the chromatographic bed are disconnected from the system so that they can be treated with discrete, discrete changes in eluent conditions.

[0033]

[0036] "Concurrently contacting sequentially" means that one or more segments of a fluidly interconnected chromatographic bed are simultaneously contacted with at least two different materials, such that the flow of one material through the interconnected segments is continuous with the flow of at least another material, and together contribute to the overall material flow over the interconnected portions of the bed. This term is meant to contrast with "stepwise contacting," in which one or more segments are contacted with different materials at different times, regardless of whether the portions of the bed are fluidly interconnected. This term is also intended to distinguish from "concurrently contacting sequentially," which is a situation in which certain segments of a chromatographic bed are not connected to (or disconnected from) each other, such that the overall material flow through the entire bed is not affected by the material flow in the disconnected portions, even though different segments of the bed may be contacted with different materials at the same moment. This latter situation arises when performing continuous separations using adsorption / desorption chromatography.

[0034]

[0037] "Amorphous fructose" means a solution containing fructose and water, where the purification of the fructose in the solution is insufficient to form crystalline fructose by fructose crystallization techniques. Most commonly, amorphous fructose is less than 95% fructose, based on dissolved solids weight.

[0035]

[0038] Returning now to the present invention, the disclosure that follows is based on the discovery of an efficient process for converting readily available, low-cost feedstocks, such as HFCS, to allulose and separating the allulose-rich product eluate from the fructose-rich raffinate eluate. In one embodiment, the method involves separating a mixture of allulose, fructose, glucose, and glucooligosaccharides (wherein the separation involves using simulated moving bed chromatography) and recovering allulose in high purity and yield. In one embodiment, the simulated moving bed ("SMB") chromatography comprises five zones. In one embodiment, the method provides for the separation of allulose reaction products of allulose in high fructose corn syrup ("HFCS"). In another embodiment, the method produces a highly enriched allulose product having a dissolved solids purity of at least 95% wt / wt. In one embodiment, the apparatus produces a highly enriched allulose product having a dissolved solids purity of at least 98% wt / wt. In another embodiment, the method produces a high-purity fructose product with a dissolved solids purity of at least 80% wt / wt, preferably at least 88% wt / wt, more preferably at least 90% wt / wt, which can be recycled to a column containing immobilized epimerase to further convert fructose to allulose in an enzymatic reaction (also referred to herein as an allulose epimerase process).In a further embodiment, the glucose obtained as a raffinate from the SMB chromatography device (also referred to as a glucose-rich fraction) can be recycled to a glucose isomerase enzymatic reaction, for example, a column containing immobilized glucose isomerase, to convert part of the glucose to fructose and produce more HFCS.In yet another embodiment, the method produces a glucose-rich fraction that can be recycled to a glucose isomerase enzymatic reaction to convert part of the glucose to fructose.

[0036]

[0039] In some embodiments, each SMB chromatography zone comprises at least one chromatography column containing a resin. In some embodiments, each column has the same dimensions and contains the same resin. The first zone may be referred to as the feed zone, the second zone may be referred to as the desorbent zone, the third zone may be referred to as the dextrose-enriched zone (or split zone), the fourth zone may be referred to as the allulose-enriched zone, and the fifth zone may be referred to as the reload zone.

[0037]

[0040] One aspect of the methods provided herein is the use of a simulated moving bed chromatographic apparatus containing a chromatographic bed material to simultaneously separate the allulose-enriched product from the fructose- and dextrose-enriched product, and the fructose-enriched product from the dextrose-enriched product.

[0038]

[0041] Simulated moving bed chromatography apparatus containing ion exchange chromatographic bed material has been used in the separation of hydrophilic products, such as basic amino acids and acidic carboxylic acids, from fermentation broths using polar ion exchange chromatographic bed materials. In these processes, the simulated moving bed apparatus is configured to perform the separation by absorption / desorption chromatography, which requires a discontinuity in connections to switch from first eluent conditions, where the desired product binds to the chromatographic bed, to second conditions, where the desired product desorbs from the bed. While these are discontinuous separation techniques, simulated moving bed apparatus are typically mounted on a carousel; therefore, by appropriate use of valves and eluent streams at different stations, the process can be operated in a continuous manner.

[0039]

[0042] While certain embodiments provided herein may use absorption / desorption separations, more advantageous embodiments employ true simulated moving-bed chromatographic separations. True simulated moving-bed chromatographic separations are continuous chromatographic separation techniques in which the fluid-phase material containing the species to be separated may be stationary, but a simulated moving-bed apparatus is used to mimic the effect that would be observed if a solid-phase chromatographic material were to move through the fluid phase. The effect would be the separation of species into different zones within the bed material based on their relative partitioning between the fluid phase and the bed material. This effect is mimicked by dividing the chromatographic bed material into fluidly interconnected sections and moving the interconnected sections in a countercurrent direction opposite to the direction of flow of the feed material (and eluent, if different from the feed material). Thus, species that preferentially partition into the moving fluid phase will preferentially migrate in one direction, while species that preferentially partition into the solid phase will migrate in the opposite direction, thus resulting in separation into different column segments representing different zones.

[0040]

[0043] When true simulated moving bed chromatography is operated in a continuous manner, with stepwise segment movement, feed input, and product withdrawal from different zones, as provided in certain embodiments, a constant concentration gradient is established between the ends of the withdrawal zones, with one end preferentially enriched in a first product species, the opposite end enriched in the unseparated mixture, and intervening zones preferentially enriched in other product species. If products from the different zones are completely removed or optional regeneration zones are used to wash or replenish segments with input eluent, the process can be run indefinitely, providing continuous separation and isolation of multiple species from the feed without the need for intervention.

[0041]

[0044] In any simulated moving bed chromatographic device, the chromatographic bed material contained within the device is conceptually divided into zones, each zone being distinguishable from other zones by the flow of fluid within the chromatographic bed material of that zone. Zones may also be distinguished, for example, by the eluent introduced or withdrawn in that zone or the primary function occurring within that zone. In certain embodiments where different fluids are applied in different zones, a gradient is established in which the content of the first fluid increases and the content of the second fluid decreases (or vice versa) in opposite directions relative to the location of the input zone.

[0042]

[0045] In a typical simulated moving bed apparatus, multiple interconnected chromatographic bed segments are arranged in a sequential series, and fluid ports are provided so that feedstock, eluent, or other mobile phase materials can be introduced or removed from any selected segment or location within the apparatus. An arrangement of valves at the top and bottom of each segment manages the flow of fluids into and out of any number of interconnected segments in the same or different zones at independently controllable flow rates. Column segments are typically arranged in a carousel-type arrangement, which circulates the column segments by circularly moving their positions in discrete steps over the course of a cycle. In this configuration, the ports in contact with the column segments at the top and bottom of each segment are fixed, and thus the column segments circulate by circularly moving relative to the fixed ports. During a complete cycle, each column segment passes through a different set of positions and fixed ports, each with a different primary function occurring. The function occurring at any given position remains constant, and thus the position of the segment conceptually designates its zone. In an alternative carousel configuration, the column segments are fixed, and the ports that contact each column segment at its top and bottom move circularly about the column segments. In a complete cycle, the movement of the ports causes each column segment to pass through different positions where different primary functions occur. The function occurring at any given position remains constant, and thus the position of the segment conceptually designates its zone.

[0043]

[0046] The number of chromatographic beds, columns, or ports thereof connected in series is unlimited. The method of the present invention can be optimized by adjusting the flow rates for fluid input and output and the timing of segment (or port) movement to improve product yield. Another variable for optimizing the method of the present invention is the number of column segments used to define successive zones. In the series, each zone can have an optimized number of column segments. Thus, while the figures provided herein each show exemplary configurations with 12, 14, or 20 column segments for ease of illustration, the method provided herein is not limited to a specific number of chromatographic devices. In the series of column segments, one or more segments define the zones. Each zone can be defined by the number of column segments into which species separation occurs when an eluent stream is introduced or removed. Thus, the method is scalable to any practical dimensions by those skilled in the art.

[0044]

[0047] In one embodiment, an allulose production facility is provided, including a simulated moving bed apparatus configured to operate any one of the processes provided herein. Such a facility may be characterized by having a simulated moving bed apparatus configured with ports that simultaneously direct a stream of feedstock containing fructose, dextrose (i.e., D-glucose), and allulose to a chromatographic bed material, and by simultaneously removing allulose, fructose, and a dextrose-enriched product in different zones of the simulated moving bed apparatus. The allulose production facility may be part of, connected to, or incorporated into a high fructose corn syrup production facility in which dextrose is converted to a mixture containing glucose and fructose by contacting a dextrose solution with glucose isomerase immobilized on a column. The allulose production facility also preferably contains an allulose epimerase immobilized on a column, which converts fructose to allulose by contacting the high fructose corn syrup with the epimerase enzyme.

[0045]

[0048] In some embodiments, the apparatus includes a water source, a sugar source, and a simulated moving bed ("SMB") chromatography column. The column may include five zones. In some embodiments, the apparatus provides for the separation of allulose reaction products from other sugars and oligosaccharides present in high fructose corn syrup ("HFCS"). In some embodiments, the apparatus produces a highly enriched allulose product having a dissolved solids purity of at least 90% wt / wt. In some embodiments, the apparatus produces a highly enriched allulose product having a dissolved solids purity of at least 95% wt / wt. In some embodiments, the apparatus produces a highly enriched allulose product having a dissolved solids purity of at least 98% wt / wt. In another embodiment, the device produces a high-purity fructose product having a dissolved solids purity of at least 80% wt / wt, preferably at least 88% wt / wt, more preferably at least 90% wt / wt, which can be recycled to a column containing immobilized epimerase for further conversion of fructose to allulose in an enzymatic reaction (also referred to herein as an allulose epimerase process). In a further embodiment, the glucose obtained as a raffinate from the SMB device can be recycled to a column containing immobilized glucose isomerase to produce more HFCS.

[0046]

[0049] FIG. 1 illustrates an exemplary system for implementing the methods provided herein. In this exemplary embodiment, a simulated moving bed chromatography apparatus 100 includes a water source 102, a mixed sugar source 104, and an SMB apparatus containing five zones: a feed zone 106, a desorbent zone 108, an allulose-enrichment zone 110, a dextrose-enrichment zone 112, and a reload zone 114. The apparatus 100 includes a plurality of chromatography column segments 1-12 connected in sequential fluid series via a conduit 116 to form a continuous chromatographic bed, each of which has a chromatographic bed material 122. The chromatographic bed material 122 is composed of an ion exchange resin (most commonly, the calcium type). Examples of suitable chromatographic bed materials include, but are not limited to, strongly acidic cationic resins. Examples of suitable strong acid cation resins include those sold by Dow Chemical Co. (Midland, Michigan) under product numbers Dowex 99 Ca / 320, Dowex 99 Ca / 310, and Dowex 99 Ca / 280; and those sold by Mitsubishi Chemical Co. (Tokyo, Japan) under the product name DIAION TM , and those sold under product numbers SK1B, SK104H, SK110, SK112 ("SK Series"), UBK530, UBK550, UBK535, and UBK555 ("UBK500 Series").

[0047]

[0050] The sugar source 104 may be used to supply a mixture 124 of allulose, fructose, and dextrose to the feeding zone 106. The mixture 124 may be produced by the enzymatic reaction of high fructose corn syrup with an immobilized epimerase to convert the fructose in the syrup to the allulose reaction product.

[0048]

[0051] As shown in a first step 140, chromatographic bed material 122 is simultaneously and sequentially contacted with (i) a mixture 124 containing allulose, fructose, and dextrose in a first zone, i.e., feed zone 106 (column segment 7 is the first column segment defining feed zone 106), and (ii) water 130 from water source 102 in a second zone, i.e., desorbent zone 108 (column segment 1 defines the first column of desorbent zone 108). Mixture 124 is introduced into chromatographic bed material 122 through inlet port 128. Water 130 is introduced into chromatographic bed material 122 through inlet port 132. These material streams together accomplish separation by adsorbent chromatography using water as the eluent.

[0049]

[0052] In one embodiment, three pumps may be utilized to form the enrichment, split, and reload streams, as further identified below, and two additional pumps may be used to push the feed and eluent (i.e., water) through the system. As shown in Figure 1, pump 126 is used to pump feed 124 into apparatus 100, and pump 134 is used to pump water 130 into apparatus 100. As shown in Figure 1, pump 138 is used to form the enrichment stream from desorbent zone 108 to allulose enrichment zone 110, pump 158 is used to form the split stream from feed zone 106 to dextrose enrichment zone 112, and pump 160 is used to form the reload stream from dextrose enrichment zone 112 to reload zone 114.

[0050]

[0053] Pump 134 pumps water 130 from inlet port 132 to force water 130 into apparatus 100 in a first direction 136. As shown in FIG. 1 , first direction 136 is the liquid-phase direction indicated in the legend under "Process Direction." Eluate from desorbent zone 108 (i.e., the second zone) is pumped in first direction 136 by pump 138 toward feed zone 110 (i.e., the first zone) to contact a mixture 124 of allulose, fructose, and dextrose. Mixture 124 is pumped in first direction 136 by pump 126. Pump 126 pumps mixture 124 into feed zone 106 (i.e., the first zone).

[0051]

[0054] 1, second direction 144 is also the stationary phase direction indicated in the legend under "Process Directionality." In one embodiment, step 142 occurs after step 140. As shown in step 142, columns 1-12 are transitioned in second direction 144. As shown in step 142, each column is moved to the location where its adjacent column was located in step 140. As shown in step 142, column 1 is then where column 12 was previously located in step 140, then column 12 is where column 11 was previously located in step 140, then column 11 is where column 10 was previously located in step 140, then column 10 is where column 9 was previously located in step 140, then column 9 is where column 8 was previously located in step 140, then column 8 is where column 7 was previously located in step 140, then column 7 is where column 6 was previously located in step 140, then column 6 is where column 5 was previously located in step 140, then column 5 is where column 4 was previously located in step 140, then column 4 is where column 3 was previously located in step 140, then column 3 is where column 2 was previously located in step 140, and then column 2 is where column 1 was previously located in step 140.

[0052]

[0055] In some embodiments, the first step 140 is followed by a second step 142, and the second step 142 is followed by a subsequent step (not shown in FIG. 1 ), with columns 1-12 transitioning multiple times until each column returns to its original location in the first step 140. In some embodiments, each step occurs over the same period of time. In the embodiment shown in FIG. 1 , upon completion of the 12th step, each column has been positioned at every column location in the apparatus 100, completing one column rotation. In some embodiments, the columns may be transitioned more than 12 times. For example, the columns may be transitioned 37 times, or three complete rotations (12 steps per rotation), such that in the 37th step, each column is in the same location as in the first step 140.

[0053]

[0056] As shown in FIG. 1 , the simulated moving bed chromatography apparatus 100 includes a plurality of column segments 1-12 connected in series and containing a chromatographic bed material 122, and simultaneously: a. a fructose, dextrose, and allulose mixture 124 is fed at a feed port location, i.e., feed inlet port 128, to contact the chromatographic bed material 122 in the column segments of a first zone, i.e., feed zone 106; b. water 130 is fed into the apparatus 100 at an eluent port location, i.e., inlet port 132, to contact the chromatographic bed material 122 in the column segments of a second zone, i.e., desorbent zone 108; and c. the water 130 flows in a first direction 136 toward the first zone, to contact the fructose, dextrose, and allulose mixture 124, and moves through the plurality of column segments 1-12 in a second direction 144 opposite to the first direction 136. (The movement of the column segments is described by the column positions in step 140 and the next step 142); d. removing a first eluate 146 enriched in allulose reaction product from a product port 148 located in a second zone, i.e., desorbent zone 108, upstream of a feed port position, i.e., inlet port 128, relative to the first direction 136; e. removing a second eluate 150 enriched in fructose from a first raffinate port position, i.e., first raffinate port 152, in a first zone, i.e., feed zone 106, downstream of the feed inlet port 128, relative to the first direction 136; f. removing a third eluate 154 enriched in dextrose (also called D-glucose) from a second raffinate port position, i.e., second raffinate port 156, in a third zone, i.e., dextrose enrichment zone 112, downstream of the first raffinate port, relative to the first direction 136.

[0054]

[0057] Allulose is preferentially distributed to the solid-phase chromatographic bed material 122 through adsorption interactions. At separate times, column segments 1-12 are simultaneously moved one step in a second direction 144, which is opposite (counterflow) to the first direction 136 of liquid phase flow. Because allulose is preferentially distributed to the solid-phase chromatographic bed material 122, repeated periodic movement of column segments 1-12 causes allulose to be preferentially moved across the solid-phase chromatographic bed material 122 in the second direction 144 toward (step 140) a product port 148 shown in column segment 2, where allulose is removed from the chromatographic bed material 122. Because the allulose reaction product 146 is removed (desorbed) from the chromatographic bed material 122 at port 148, column segments 3-6 between port 148 and the feed inlet port 128 are designated as an allulose-enrichment zone 110. The allulose reaction product 146 moves in the second direction 144 with the phase chromatographic bed material 122, while the fructose and dextrose continue to move in the first direction 136 with the mobile liquid phase, and a fructose-enriched eluate 150 is removed from a first raffinate port 152 shown in column 9 (step 140), and a dextrose-enriched eluate 154 is removed from a second raffinate port 156 shown in column 11 (step 140).

[0055]

[0058] FIG. 2 illustrates an exemplary system for implementing the methods provided herein. The method is a schematic flow diagram of simulated moving bed chromatography according to an embodiment of the present invention. In one embodiment, simulated moving bed chromatography apparatus 200 includes column segments 1-14. Apparatus 200 differs from apparatus 100 shown in FIG. 1 in that apparatus 200 has two additional column segments. Apparatus 200 has five column segments in allulose enrichment zone 110, while apparatus 100 has four column segments in allulose enrichment zone 110. Apparatus 200 has three column segments in desorbent zone 108, while apparatus 100 has two column segments in desorbent zone 108.

[0056]

[0059] FIG. 3 illustrates an exemplary system for implementing the methods provided herein. The method is a schematic flow diagram of simulated moving bed chromatography according to an embodiment of the present invention. In one embodiment, simulated moving bed chromatography apparatus 300 includes column segments 1-20. Apparatus 300 differs from apparatus 100 shown in FIG. 1 in that the apparatus has eight additional column segments. Apparatus 300 has four column segments in feed zone 106, while apparatus 100 has three column segments in feed zone 106. Apparatus 300 has seven column segments in allulose enrichment zone 110, while apparatus 100 has four column segments in allulose enrichment zone 110. Apparatus 300 has four column segments in desorbent zone 108, while apparatus 100 has two column segments in desorbent zone 108. Apparatus 300 has three column segments in dextrose enrichment zone 112, while apparatus 100 has two column segments in dextrose enrichment zone 112. Apparatus 300 has two column segments in reload zone 114, while apparatus 100 has one column segment in reload zone 114.

[0057]

[0060] In an alternative embodiment, each zone has a corresponding inlet valve for the inlet of water from the water source 102, an inlet valve for the inlet of the sugar feed from the sugar source 104, and an inlet valve for the inlet of the feed from the immediately upstream column in the loop. The feed to the first column of each zone can be varied to simulate a moving bed. In Figure 1, the arrows identified as "stationary phase" pointing from right to left indicate that by opening and closing the inlet valves and varying the type of feed to the first column of each zone in a specific manner, a simulated moving bed is obtained that rotates in a circular clockwise direction, i.e., from left to right as shown in Figure 1, even though the columns and the resin within them do not move.

[0058]

[0061] The outlet stream from each column enters the top of the next downstream column, and all columns are connected in a flow loop.

[0059]

[0062] The sugar source 104 is allulose (A x ), fructose (F x ) and dextrose (D x ) mixture. x ) is also known as D-glucose. Allulose is also known as psicose.

[0060]

[0063] Figure 4 is a flow diagram illustrating the production of allulose from glucose in a corn processing facility in accordance with an embodiment of the present invention. Process 400 includes an allulose purification process 402 that includes a ternary chromatographic separation process 404 using simulated moving bed (SMB) chromatography as described hereinabove and illustrated in Figures 1-3, which simultaneously results in the separation of an allulose-enriched stream 406 from a fructose- and glucose-enriched stream (not shown in Figure 4) and the separation of a fructose-enriched stream 408 from a glucose-enriched stream 410. Those skilled in the art, having the benefit of this disclosure, will recognize that ternary chromatographic process 404 can be implemented using any suitable apparatus, such as apparatus 100 of Figure 1, apparatus 200 of Figure 2, or apparatus 300 of Figure 3. In addition to the allulose purification process 402, the overall process 400 also includes a dextrose purification process 470 in which dextrose is produced by liquefaction of corn starch; a fructose purification process 432 in which dextrose is converted to HFCS having a fructose content of about 42% DS; and a fructose enrichment process 434 in which a portion of the HFCS is enriched to about 90% dissolved solids fructose, and a portion of the HFCS undergoes the allulose purification process 402.

[0061]

[0064] 4, the allulose purification process 402 can include an allulose epimerase process 412, an ion exchange process 414, and a filtration process 416. In the allulose epimerase process 412, an HFCS feedstock 418 is placed in contact with a suitable epimerase enzyme to convert a portion of the fructose in the HFCS to allulose. In this embodiment, in the allulose epimerase process 412, a mixture 420 exiting the allulose epimerase process 412 includes fructose, dextrose, and allulose resulting from the enzymatic reaction of the HFCS feedstock 418 with the immobilized epimerase to convert a portion of the fructose in the syrup to an allulose reaction product. The immobilized epimerase used in the allulose epimerase process 412 can be any suitable epimerase that converts fructose to allulose. Such suitable epimerases include, but are not limited to, variants of Burkholderia multivorans CGD1 xylose isomerase engineered to have approximately 1.5- to 2-fold improved catalytic activity relative to the parent enzyme, as disclosed in PCT / US19 / 45400 (filed August 7, 2019), which is incorporated herein by reference. In ion exchange process 414 and filtration process 416, mixture 420 containing fructose, dextrose, and allulose reaction products can be processed to obtain filtered mixture 422 containing fructose, dextrose, and allulose.

[0062]

[0065] In one embodiment, food-grade magnesium sulfate can be added to the HFCS feedstock 418 before it is conveyed to the allulose epimerase process 412. The magnesium sulfate acts as a stabilizer for the epimerase. In an ion exchange process 414, magnesium salts are removed from the mixture 420 containing fructose, dextrose, and allulose. In a filtration process 416, entrained resin beads can be removed from the mixture 420 to obtain a filtered mixture 422 containing fructose, dextrose, and allulose reaction products.

[0063]

[0066] 4, the filtered mixture 422 containing the fructose, dextrose, and allulose reactants is conveyed to a ternary chromatography process 404. In the ternary chromatography process 404, the filtered mixture 422 containing fructose, dextrose (also known as D-glucose), and allulose is separated to produce an allulose-enriched stream 406, a fructose-enriched stream 408, and a glucose-enriched stream 410. In an embodiment of the invention, the fructose-enriched stream 408 has a larger volume than the glucose-enriched stream 410.

[0064]

[0067] As shown in FIG. 4 , additional processes may be downstream of the ternary chromatography process 404. The allulose-enriched stream 406 may undergo further processing to obtain allulose 426 from the allulose-enriched stream 406. Those skilled in the art, having the benefit of this disclosure, will recognize that any suitable processing may be used to obtain allulose 426 from the allulose-enriched stream 406. In certain embodiments, such processing may include an evaporation process 424 to evaporate water from the allulose-enriched stream 406, ion exchange processing 428, and an evaporation process 430 following the ion exchange processing 428 to further evaporate water from the allulose-enriched stream to obtain allulose 426. In certain embodiments, the evaporation process 424 may be used to increase the dry solids of the allulose-enriched stream 406 from about 10-15% DS to about 50% DS. The ion exchange process 428 may be used to remove color so that the allulose-enriched stream 406 is clear or more transparent. An evaporation process 430 can be used to increase the dry solids of the allulose-enriched stream 406 from about 50% DS to about 77% DS to obtain allulose 426. Allulose 426 can be characterized as allulose syrup or 90 allulose @ 77 DS.

[0065]

[0068] 4, the fructose-enriched stream 408 can be recycled to an enzymatic reaction of high fructose corn syrup, i.e., allulose epimerase process 412, where the recycled fructose can be converted to allulose. An advantage of recycling the fructose-enriched steam 408 (also referred to herein as the "enriched fructose fraction") in this manner is that the enriched fructose fraction does not need to undergo evaporation prior to the enzymatic reaction to convert the fructose in the fraction to allulose.

[0066]

[0069] As shown in Figure 4, the glucose-enriched stream 410 (also referred to herein as the "enriched dextrose fraction") can be recycled as a feedstock for an upstream fructose purification process 432, where the dextrose (also called D-glucose) is converted to HFCS. In one embodiment, the upstream fructose purification process 432 can be followed by a fructose enrichment process 434 (labeled "90 Fructose Purification" in Figure 4).

[0067]

[0070] As shown in Figure 4, fructose enrichment process 434 can be used to convert the HFCS stream 448 from fructose purification process 432 into a more highly enriched fructose stream. Fructose enrichment process 434 can include chromatography process 436, evaporation process 438, ion exchange process 440, and evaporation process 442 to produce an enriched fructose feedstock 418 that is then used as a feedstock for allulose purification process 402. Those skilled in the art, having the benefit of this disclosure, will recognize that chromatography process 436, evaporation process 438, ion exchange process 440, and evaporation process 442 are processes that can be used to produce enriched fructose 446, such as 90HFCS@77 dry solids ("DS"), and that filtration process 444 can also be used as a polishing step after evaporation process 442, as shown in Figure 4, to produce filtered 90HFCS@77DS that can be used in certain commercial applications. Chromatography process 436 may be a two-way chromatography process used to separate fructose from dextrose. Evaporation process 438 may be used to increase the dry solids in the fructose stream resulting from chromatography process 436 from about 10-15% DS to about 50% DS. Ion exchange process 440 may be used to remove color from the fructose stream. Evaporation process 442 may be used to increase the dry solids in the fructose stream from about 50% DS to obtain HFCS feedstock 418, which may be characterized as 90% HFCS @ 77% DS.

[0068]

[0071] As shown in FIG. 4, fructose purification process 432 may include a glucose isomerase enzyme process 450, an evaporation process 452, a carbon process 454, a filtration process 456, and an ion exchange process 458 to produce a fructose stream 448 that is then used as a feedstock for fructose enrichment process 434. In glucose isomerase enzyme process 450, dextrose (i.e., D-glucose) is converted to fructose. Evaporation process 452 may be used to increase the dry solids in the fructose-containing stream from glucose isomerase enzyme process 450. Carbon process 454 may be a process that uses granular carbon to remove flavors and odors. Filtration process 456 may be used to remove undesirable particulates. Ion exchange process 458 may be used to remove color. Fructose stream 448 may be conveyed from ion exchange process 458 to chromatography process 438 of fructose purification process 434.

[0069]

[0072] As shown in Figure 4, fructose purification process 432 can include a filtration process 460 and an evaporation process 462 to convert stream 464 from ion exchange process 458 to produce 42HFCS@71DS, identified as stream 466 in Figure 4. Stream 466, i.e., 42HFCS@71DS, can be used in certain commercial applications.

[0070]

[0073] As previously discussed, glucose-rich stream 410 can be recycled as a feedstock for an upstream fructose purification process 432, where dextrose (also known as D-glucose) is converted to fructose for the production of HFCS. In the embodiment shown in Figure 4, glucose-rich stream 410 can be conveyed to ion exchange process 468 of dextrose purification process 470. Those skilled in the art, having the benefit of this disclosure, will recognize that dextrose purification process 470 can convert corn starch 472 into dextrose feedstock stream 474, which can then be converted to fructose stream 448 as described above.

[0071]

[0074] Those skilled in the art with the benefit of this disclosure will recognize that dextrose purification process 470 can include, in addition to and upstream of ion exchange process 468 already discussed herein, converter process 476, liquefaction process 478, saccharification process 480, filtration process 482, evaporation process 484, carbon process 486, and filtration process 488. Ion exchange process 468 can be used to remove ions that may impair the enzyme in glucose isomerase enzyme process 450. Converter process 476 can convert corn starch 472 to a globular liquifactant. Liquefaction process 478 can include the addition of water, and acid to further solubilize the starch, and saccharification process 480 can include alpha amylase and glucoamylase to convert the solubilized starch to dextrose. Filtration process 482 can be used to remove unsolubilized starch. Evaporation process 484 can be used to increase dry solids. Carbon process 486 may be a process that uses granular carbon to remove flavors and odors. Filtration process 488 may be used to remove unwanted particulates from the process stream before it is conveyed to ion exchange process 468.

[0072]

[0075] As described above, embodiments of the present invention include SMB chromatography, a three-way (also referred to as "ternary") chromatography that separates allulose from fructose and glucose, and then separates fructose from glucose. In a two-way chromatography process in which allulose is separated from a mixture of fructose and D-glucose (i.e., dextrose), and the mixture of fructose and D-glucose is recycled to an ion exchange process 468 and then to a glucose isomerase enzyme process 450, it has been found that reprocessing the mixture of fructose and D-glucose, which is primarily D-glucose, reduces glucose isomerase productivity due to the high fructose concentration in the mixture. This reduction in glucose isomerase productivity has been found to substantially reduce the efficiency of processing corn starch to HFCS and HFCS to allulose.

[0073]

[0076] It has been discovered that the productivity of glucose isomerase enzymatic process 450 can be increased by separating fructose-rich stream 408 from glucose-rich stream 410 using three-way chromatography, SMB chromatography, and recycling only glucose-rich stream 410 to ion exchange process 468 and then to glucose isomerase enzymatic process 450 as part of dextrose feed stream 474. By using three-way chromatography, SMB chromatography, a stream containing greater than 85% DS weight fructose by weight (i.e., fructose-rich stream 408) is obtained, and another stream containing greater than 90% DS weight dextrose by weight (i.e., glucose-rich stream 410) is obtained. Fructose-rich stream 408 has a larger volume than glucose-rich stream 410. By recycling the fructose-rich stream 408 directly back to the allulose epimerase process 412 and recycling only the glucose-rich stream 410 to the ion exchange process 468 and then to the glucose isomerase enzyme process 450 as part of the dextrose feed stream 474, the overall process is much more efficient. In one embodiment, the fructose-rich stream 408 is the same as the fructose-rich eluate 150 shown in Figures 1-3 and described above.

[0074]

[0077] In summary, according to embodiments of the present invention, dextrose purification process 470 converts corn starch into dextrose feed stream 474, fructose purification process 432 converts dextrose feed stream 474 into HFCS stream 448, fructose enrichment process 434 converts HFCS stream 448 into 90% enriched fructose feed stream 418, and allulose purification process 402 involves converting enriched fructose feed stream 418 into allulose 426 and glucose enriched stream 410 and recycling fructose enriched stream 408 (with unconverted fructose) to allulose epimerase process 412 for further conversion of fructose to allulose.

[0075]

[0078] The present invention will be more specifically explained by the following examples. [Example]

[0076]

[0079] Example 1

[0080] The pilot SMB was plumbed in the configuration shown in Figure 1. The resin used was styrene-DVB, a resin for the chromatographic separation of sugars with strong acid cations and sulfonic acid functional groups in a gel matrix, specifically Dowex Monosphere TM The column volume was measured to be approximately 331 mL / column. Three pumps were utilized to create the enrichment, split, and reload streams, and two additional pumps were used to push the feed and desorbent through the system.

[0077]

[0081] To test this experiment, a pre-weighed container of allulose syrup (allulose concentration was 18-20% based on dry solids) made from HFCS90 was used as the feed, and DI water was used as the desorbent. The process described above was performed. Specifically, the system was allowed to reach equilibrium by rotating the column segments 37 times, i.e., three complete revolutions (12 steps per revolution), so that each column was in the same location in the 37th step as it was in the first step.

[0078]

[0082] Once the system reached equilibrium, the feed and desorbent were connected to the inlets of the feed and desorbent pumps, and three outlets were set up for collection in pre-weighed containers. The system was then run for 55.2 minutes, after which all containers were weighed and samples were sent analytically for analysis. Six steps of 9.2 minutes per step were performed during the 55.2 minute period. The bed volume in Table 1 below refers to the volume of resin per column.

[0079] [Table 1]

[0080]

[0084] In Table 1 above, (i) "Feed" is the flow of the allulose reaction product, fructose, and dextrose mixture 124 into the first zone, i.e., feed zone 106; (ii) "Desorption" is the flow of water 130 from the water source 102 to the second zone, i.e., desorbent zone 108; (iii) "Enrichment" is the flow from the desorbent zone 108 to the allulose enrichment zone 110; (iv) "Reload" is the flow from the dextrose enrichment zone 112 to the reload zone 114; and (v) "Split" is the flow from the feed zone 106 to the dextrose enrichment zone 112.

[0081]

[0085] In Table 1, f1 through f5 are flow rates, where f1 is the flow rate through the desorbent zone 108, f2 is the flow rate through the allulose enrichment zone 110, f3 is the flow rate through the feed zone 106, f4 is the flow rate through the dextrose enrichment zone 112, and f5 is the flow rate through the reload zone 114.

[0082]

[0086] Example 2

[0087] Table 2 shows the performance of a 12-column bed configuration ( FIG. 1 ), a 14-column bed configuration ( FIG. 2 ), and a 20-column bed configuration ( FIG. 3 ), all using a three-way chromatography process 404 ( FIG. 4 ), with a fructose-enriched stream 408 recycled to the allulose epimerase process 412 and a glucose-enriched stream 410 recycled to the ion exchange process 468 (upstream of the fructose purification process 432). In Table 2, "dsb" means dry solids basis.

[0083]

[0088] In some embodiments, dextrose (also called D-glucose) can be removed from the allulose purification process 402 as a glucose-enriched stream 410. In some embodiments, the glucose-enriched stream 410 is the same as the dextrose-enriched eluate 154 shown in Figures 1-3. The process can be performed in which the raffinate, i.e., the dextrose-enriched eluate 154, can be greater than 50% by weight, more preferably at least about 87% by weight dextrose (also called D-glucose) (and closer to 85% by weight dextrose, not counting DP). By removing dextrose in this manner using three-way chromatography, the dextrose-enriched eluate 154 can be sufficiently pure to be recycled as a glucose-enriched stream 410 for enzymatic conversion of dextrose to fructose. By recycling the fructose-rich stream 408 directly back to the allulose epimerase process 412 and recycling only the glucose-rich stream 410 to the ion exchange process 468 and then to the glucose isomerase enzyme process 450, the overall process is much more efficient than a process using dual chromatography in which all of the raffinate, after removal of allulose, is recycled upstream of the ion exchange process 468 and then to the glucose isomerase enzyme process 450.

[0084]

[0089] Table 2 shows that the 20-column bed configuration (FIG. 3), together with the three-way chromatography process 404 of the present invention in which 90% HFCS was used as the feedstock to the allulose epimerase process, resulted in a raffinate stream (i.e., glucose-enriched stream 410) of 2.9 ml / min. As shown in Table 2, the 10-column bed configuration using the two-way chromatography process in which highly purified dissolved crystalline fructose was used as the feedstock to the allulose epimerase process resulted in a raffinate stream of 15.6 ml / min. Thus, the 20-column bed configuration (FIG. 3), together with the three-way chromatography process 404, had over 81% less raffinate than the 10-column bed with the two-way chromatography process.

[0085]

[0090] As shown in Table 2, the 14-column bed configuration (FIG. 2), in conjunction with the three-way chromatography process 404 of the present invention in which 90% HFCS was used as the feedstock to the allulose epimerase process, resulted in a 3.0 ml / min raffinate stream (i.e., glucose-enriched stream 410), and therefore, more than 80% less raffinate than the 15.6 ml / min raffinate obtained from using a 10-column bed configuration in conjunction with a two-way chromatography process in which highly purified dissolved crystalline fructose was used as the feedstock to the allulose epimerase process.

[0086]

[0091] As shown in Table 2, the 12-column bed configuration (FIG. 1), in conjunction with the three-way chromatography process 404 of the present invention in which 90% HFCS was used as the feedstock to the allulose epimerase process, resulted in a 7.0 ml / min raffinate stream (i.e., glucose-enriched stream 410), and therefore, more than 55% less raffinate than the 15.6 ml / min raffinate obtained from using a 10-column bed configuration in conjunction with a two-way chromatography process in which highly purified dissolved crystalline fructose was used as the feedstock to the allulose epimerase process.

[0087] [Table 2]

[0088]

[0093] As described above, the invention disclosed herein provides a process in which three-way SMB chromatography is used to produce separate allulose-rich, fructose-rich, and dextrose-rich streams from a sugar solution. According to an embodiment of the present disclosure, allulose syrup and HFCS can be produced from the same separation system. The process can produce 85%+ HFCS and 90% allulose syrup. Operating below these parameters can result in allulose syrup that is unsuitable for most commercial applications.

[0089]

[0094] Those skilled in the art, with the knowledge gained from this disclosure, will recognize that various changes can be made to the disclosed processes to achieve these and other advantages without departing from the scope of the present disclosure. Accordingly, it should be understood that features of the present disclosure are susceptible to modification and / or substitution. The specific embodiments illustrated and described herein are for illustrative purposes only and do not limit the invention, which is set forth in the claims.

Claims

1. A process for purifying allulose, comprising contacting a chromatographic bed material with water and a mixture of allulose, amorphous fructose, and glucose, and separating an allulose-rich fraction from a fructose- and glucose-rich fraction by adsorbent chromatography on the chromatographic bed material, and simultaneously separating a fructose-rich fraction from a glucose-rich fraction by adsorbent chromatography on the chromatographic bed material; the chromatographic bed material is contained within a simulated moving bed apparatus; The simulated moving bed apparatus comprises a plurality of movable column segments connected in sequential fluid series, and comprises, in order, an eluent port for introducing water into the simulated moving bed apparatus to contact the chromatographic bed material, a product port for removing the allulose-rich fraction from the simulated moving bed apparatus, a feed port for introducing a mixture of allulose, fructose and glucose into the simulated moving bed apparatus to contact the chromatographic bed material, a first raffinate port for removing the fructose-rich fraction from the simulated moving bed apparatus, and a second raffinate port for removing the glucose-rich fraction from the simulated moving bed apparatus.

2. 2. The process of claim 1, wherein the contacting of the chromatographic bed material with the water and the mixture of allulose, fructose and glucose using a pseudo-adsorbent chromatographic separation is carried out sequentially with removal of the allulose-rich fraction, removal of the fructose-rich fraction, and removal of the glucose-rich fraction.

3. 2. The process of claim 1, wherein the mixture of allulose, fructose and glucose is contacted with the chromatographic bed material in a first zone and the allulose is removed from a second zone that is different from the first zone and upstream of the first zone with respect to the direction of water flow.

4. 4. The process of claim 3, further comprising contacting the chromatographic bed material with the water in the second zone.

5. 5. The process of claim 4, further comprising removing the fructose-rich fraction from a third zone downstream of the first zone relative to the direction of water flow.

6. 6. The process of claim 5, further comprising recycling the fructose-rich fraction after removal from the first zone and contacting it with an allulose epimerase to convert the fructose to allulose, wherein the allulose epimerase is located upstream of the chromatographic bed material.

7. 7. The process of claim 6, further comprising removing the glucose-rich fraction from a fourth zone, the fourth zone being located downstream of the third zone with respect to the direction of flow of the water and the mixture of allulose, fructose and glucose.

8. 8. The process of claim 7, further comprising recycling the glucose-rich fraction after removal from the fourth zone and contacting it with a glucose isomerase enzyme to convert the glucose to fructose, wherein the glucose isomerase enzyme is located upstream of the allulose epimerase.

9. 9. The process of claim 8, wherein the fructose-rich fraction has a larger volume than the dextrose-rich fraction.

10. 1. A simulated moving bed apparatus comprising a plurality of column segments connected in series and containing chromatographic bed material, a. feeding a mixture of fructose, dextrose, and allulose at a feed port location to contact the chromatographic bed material in a column segment of a first zone; b. feeding water into the device at an eluent port location to contact the chromatographic bed material in the column segment of a second zone; c) flowing the water in a first direction toward the first zone to contact the fructose, dextrose, and allulose mixture and moving the plurality of column segments in a second direction opposite the first direction; d. removing the allulose-rich first eluate from a product port located in the second zone upstream of the feed port location relative to the first direction; e. removing a fructose-rich second eluate from a first raffinate port location in the first zone downstream of the feed port in the first direction; and f. removing a dextrose-rich third effluent from a second raffinate port location in a third zone downstream of said first raffinate port with respect to said first direction.

2. The process of claim 1, comprising simultaneously:

11. a. a vessel containing an immobilized allulose epimerase, wherein when high fructose corn syrup is contacted with the immobilized allulose enzyme, the allulose epimerase converts fructose to allulose; b. A simulated moving bed apparatus comprising a plurality of column segments connected in series and containing chromatographic bed material, the plurality of column segments being located downstream of a vessel containing the immobilized allulose epimerase; c. a feed port in a column segment of a first zone for receiving a mixture of fructose, dextrose, and allulose from a vessel containing the immobilized allulose epimerase and contacting the chromatographic bed material; d. an eluent port in the column segment of the second zone for receiving water to contact the chromatographic bed material; e. the simulated moving bed apparatus is configured to flow the water in a first direction toward the first zone to contact the fructose, dextrose, and allulose mixture and to move the plurality of column segments in a second direction opposite the first direction; d. a product port disposed in the second zone upstream of the feed port location relative to the first direction for removing the allulose-rich first eluate; e. a first raffinate port for removing a fructose-rich second eluate, the first raffinate port being located in the first zone downstream of the feed port with respect to the first direction; and f. a second raffinate port for removing a third dextrose-rich eluate, the second raffinate port being located in a third zone downstream of the first raffinate port with respect to the first direction; 11. A system for carrying out the process of claim 10, comprising:

12. 12. The system of claim 11, further comprising an allulose production apparatus comprising a vessel containing an immobilized glucose isomerase enzyme configured to convert glucose to fructose, the allulose production system configured to recycle the glucose-rich third eluate to the allulose production apparatus.

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