A device for preparing glucosyl-steviol glycoside in multiple enzyme steps

Abstract

The utility model discloses a kind of multi-enzyme step-by-step preparation glucosyl steviol glycoside's device, it is related to glucose steviol glycoside production technical field, the inlet of first reaction tank is respectively communicated with raw material tank, dextrin tank, cyclodextrin glucoside transferase tank and first sodium hydroxide solution tank, the outlet of first reaction tank is communicated with hydrolysis tank, the outlet of hydrolysis tank is communicated with second reaction tank, the inlet of second reaction tank is respectively communicated with sucrose tank, sucrose synthase tank, uridine diphosphate tank, UGT glycosyltransferase tank and lye tank, the outlet of second reaction tank is communicated with concentration tank by pipeline, the lower outlet of concentration tank is communicated with drier, the outlet of drier is communicated with glucosyl steviol glycoside tank. Make residual substrate conversion is relatively more optimal steviol glycoside in taste, improve the taste, quality and yield of product.

Description

Technical Field

[0001] This utility model relates to the field of glucosylstevioside production technology, specifically to an apparatus for the stepwise preparation of glucosylstevioside using multiple enzymes. Background Technology

[0002] Glucosylstevioside is a natural sweetener, also known as steviol glycoside. It consists of two components, glucose and steviol glycoside, and is characterized by high sweetness, low calories, and non-cavity-causing properties. It is widely used in the food and beverage industry.

[0003] Currently, the enzymatic preparation of glucosylsteviosides mainly has the following problems: First, the content of high-content rebaudioside A or steviol monoglycosides in unreacted glycosides is greater than 4%, which cannot meet the national standard requirements and limits its market application as a single product; Second, the amount of other steviol glycosides added in combination is very large, which causes significant changes to the properties of the product. Summary of the Invention

[0004] The technical problem to be solved by this utility model is to provide an apparatus for the stepwise preparation of glucosylstevioside by multiple enzymes, which provides glucosylstevioside with good quality and taste, in order to address the shortcomings of the existing technology.

[0005] To solve the above-mentioned technical problems, the technical solution of this utility model is as follows:

[0006] An apparatus for the stepwise preparation of glucosylsteviosides using multiple enzymes includes a first reaction vessel. The inlet of the first reaction vessel is connected via pipes to a raw material vessel, a dextrin vessel, a cyclodextrin glucoside transferase vessel, and a first sodium hydroxide solution vessel. The outlet of the first reaction vessel is connected via pipes to a hydrolysis vessel. The inlet of the hydrolysis vessel is connected via pipes to a saccharifying enzyme vessel. The outlet of the hydrolysis vessel is connected via pipes to a second reaction vessel. The inlet of the second reaction vessel is connected via pipes to a sucrose vessel, a sucrose synthase vessel, a uridine diphosphate vessel, a UGT glycosyltransferase vessel, and a second sodium hydroxide solution vessel.

[0007] The outlet of the second reaction vessel is connected to a concentration tank via a pipeline, the lower outlet of the concentration tank is connected to a dryer via a pipeline, and the outlet of the dryer is connected to a glucosylstevioside tank.

[0008] As an improved technical solution, the outlet of the second reaction vessel is connected to an adsorption resin column via a pipeline, the inlet of the adsorption resin column is connected to a first ethanol tank, a second ethanol tank, and a third ethanol tank via pipelines, the outlet of the adsorption resin column is connected to an eluent tank via a pipeline, and the outlet of the eluent tank is connected to a concentration tank via a pipeline.

[0009] As an improved technical solution, the inlet of the adsorption resin column is connected to a purified water tank via a pipeline.

[0010] As an improved technical solution, the outlet of the adsorption resin column is connected to a water-top alcohol tank via a pipeline, and the outlet of the water-top alcohol tank is connected to the concentration tank via a pipeline.

[0011] As an improved technical solution, the top outlet of the concentration tank is connected to an ethanol recovery tank via a pipeline, and the outlet of the ethanol recovery tank is connected to the ethanol tank via a pipeline.

[0012] The outlet of the second reaction vessel is connected to a plate and frame filter press via a pipeline, and the liquid phase outlet of the plate and frame filter press is connected to the adsorption resin column via a pipeline.

[0013] As an improved technical solution, the liquid phase outlet of the plate and frame filter press is connected to an ultrafiltration membrane device via a pipeline, and the clear liquid outlet of the ultrafiltration membrane device is connected to the adsorption resin column via a pipeline.

[0014] As an improved technical solution, the first reaction vessel is equipped with a first online pH sensor, and the outlet of the first sodium hydroxide solution vessel is equipped with a first automatic shut-off valve. The first online pH sensor and the first automatic shut-off valve are interlocked to the control system.

[0015] As a preferred technical solution, the second reaction vessel is equipped with a second online pH sensor, and the outlet of the second sodium hydroxide solution vessel is equipped with a second automatic shut-off valve. The second online pH sensor and the second automatic shut-off valve are interlocked to the control system.

[0016] As a preferred technical solution, the outlet of the raw material tank is connected to a crushing tank via a pipeline, the outlet of the crushing tank is connected to a dryer via a pipeline, and the outlet of the dryer is connected to the first reaction tank via a pipeline.

[0017] Due to the adoption of the above technical solution, the beneficial effects of this utility model are:

[0018] This invention discloses a multi-enzyme stepwise preparation apparatus for glucosylstevioside, comprising a first reaction vessel. The inlet of the first reaction vessel is connected via pipes to a raw material tank, a dextrin tank, a cyclodextrin glucoside transferase tank, and a first sodium hydroxide solution tank. The outlet of the first reaction vessel is connected via pipes to a hydrolysis tank. The inlet of the hydrolysis tank is connected via pipes to a saccharifying enzyme tank. The outlet of the hydrolysis tank is connected via pipes to a second reaction vessel. The inlet of the second reaction vessel is connected via pipes to a sucrose tank, a sucrose synthase tank, a uridine diphosphate tank, a UGT glycoside transferase tank, and a second sodium hydroxide solution tank. The outlet of the second reaction vessel is connected via pipes to a concentration tank. The lower outlet of the concentration tank is connected via pipes to a dryer. The outlet of the dryer is connected to a glucosylstevioside tank. Rebaudioside A is used as the raw material, β-cyclodextrin or maltodextrin as the excipient, and cyclodextrin glucoside transferase (…) is added… A glycosylation reaction was carried out using 3.0L of rebaudioside A. The pH was adjusted to 5.5-5.8 with sodium hydroxide to obtain a glucosylstevioside solution with rebaudioside A as the substrate. Then, the solution was hydrolyzed by a saccharifying enzyme to obtain low-glycosylated chain-length glucosylstevioside. This solution was then reacted with sucrose, UGT glycosyltransferase UGT76G4 (derived from Chinese patent CN115094074B), sucrose synthase (derived from sucrose synthase AtSuSy in Chinese patent CN119220516A), and uridine diphosphate glucose. The pH was adjusted to 7-8 to further convert unreacted rebaudioside A in the solution into rebaudioside I. After ethanol analysis, the solution was concentrated and dried to obtain glucosylstevioside. This process transformed the residual substrate into a steviol glycoside with a relatively better taste, improving the taste and quality of the product.

[0019] The outlet of the second reaction vessel of this invention is connected to an adsorption resin column via a pipeline. The inlet of the adsorption resin column is connected to a first ethanol tank, a second ethanol tank, and a third ethanol tank via pipelines. The outlet of the adsorption resin column is connected to an eluent tank via a pipeline, and the outlet of the eluent tank is connected to a concentration tank via a pipeline. First, elution is performed using 10-15% ethanol from the first ethanol tank to remove residual dextrin, monosaccharides, and proteins. Then, elution is performed using 20-30% ethanol from the second ethanol tank to remove weakly adsorbed impurities such as highly glycosylated glycosides. Finally, elution is performed using 60-70% ethanol from the third ethanol tank to obtain the target product, glucosylstevioside. This gradient elution improves the purity of the product and enhances its taste.

[0020] The inlet of the adsorption resin column is connected to a purified water tank via a pipeline. The purified water can be used to clean the adsorption resin column, removing residual impurities, ensuring stable adsorption performance, extending its service life, and reducing equipment maintenance costs. The outlet of the adsorption resin column is connected to a water-to-alcohol tank via a pipeline, and the outlet of the water-to-alcohol tank is connected to the concentration tank via a pipeline. The purified water can be used to top-wash the desorbed adsorption resin column, and then concentrated to obtain glucosylstevioside, improving the product yield.

[0021] The top outlet of the concentration tank is connected to an ethanol recovery tank via a pipeline, and the outlet of the ethanol recovery tank is connected to the ethanol tank via a pipeline. This achieves ethanol recycling again, further reducing production costs and environmental pollution.

[0022] The outlet of the second reaction vessel is connected to a plate and frame filter press via a pipeline. The liquid phase outlet of the plate and frame filter press is connected to the adsorption resin column via a pipeline to remove insoluble substances such as proteins, obtaining a filtered clear liquid. The liquid phase outlet of the plate and frame filter press is connected to an ultrafiltration membrane device via a pipeline. The ultrafiltration membrane device has a molecular weight cutoff of 3-10 kDa. The clear liquid outlet of the ultrafiltration membrane device is connected to the adsorption resin column via a pipeline for further impurity removal, improving the purity of the product.

[0023] The first reaction vessel is equipped with a first online pH sensor, and the outlet of the first sodium hydroxide solution tank is equipped with a first automatic shut-off valve. The first online pH sensor and the first automatic shut-off valve are interlocked to the control system. The second reaction vessel is equipped with a second online pH sensor, and the outlet of the second sodium hydroxide solution tank is equipped with a second automatic shut-off valve. The second online pH sensor and the second automatic shut-off valve are interlocked to the control system. The control system adjusts the opening and closing of the first and second automatic shut-off valves based on the values ​​of the first and second online pH sensors to ensure that the pH value of the materials in the first and second reaction vessels remains stable, which is conducive to the reaction.

[0024] The outlet of the raw material tank is connected to a crushing tank via a pipeline, the outlet of the crushing tank is connected to a dryer via a pipeline, and the outlet of the dryer is connected to the first reaction tank via a pipeline. By crushing the raw material rebaudioside A into small particles of uniform size, the reaction efficiency and yield are improved. Attached Figure Description

[0025] The present invention will be further described below with reference to the accompanying drawings and embodiments.

[0026] Figure 1 This is a structural schematic diagram of an embodiment of the present utility model;

[0027] The components are as follows: 1. First reaction vessel; 2. Raw material vessel; 3. Dextrin vessel; 4. Cyclodextrin glucosyltransferase vessel; 5. First sodium hydroxide solution vessel; 6. Hydrolysis vessel; 7. Saccharifying enzyme vessel; 8. Second reaction vessel; 9. Sucrose vessel; 10. Sucrose synthase vessel; 11. Uridine diphosphate vessel; 12. UGT glycosyltransferase vessel; 13. Second sodium hydroxide solution vessel; 14. Concentrator; 15. Dryer; 16. Glucosyl steviol glycoside vessel; 17. Adsorption resin column; 18. First ethanol vessel; 19. Second ethanol vessel; 20. Third ethanol vessel; 21. Eluent vessel; 22. Purified water vessel; 23. Water-to-ethanol vessel; 24. Ethanol recovery vessel; 25. Plate and frame filter press; 26. Ultrafiltration membrane device; 27. First online pH sensor; 28. First automatic shut-off valve; 29. ​​Second online pH sensor; 30. Second automatic shut-off valve; 31. Crushing vessel; 32. Dryer. Detailed Implementation

[0028] The present invention will be further described below with reference to the accompanying drawings and embodiments.

[0029] like Figure 1 As shown, an apparatus for the stepwise preparation of glucosylstevioside using multiple enzymes includes a first reaction vessel 1. The inlet of the first reaction vessel 1 is connected via pipes to a raw material tank 2, a dextrin tank 3, a cyclodextrin glucoside transferase tank 4, and a first sodium hydroxide solution tank 5. The outlet of the first reaction vessel 1 is connected via a pipe to a hydrolysis tank 6. The inlet of the hydrolysis tank 6 is connected via a pipe to a saccharifying enzyme tank 7. The outlet of the hydrolysis tank 6 is connected via a pipe to a second reaction vessel 8. The inlet of the second reaction vessel 8 is connected via pipes to a sucrose tank 9, a sucrose synthase tank 10, a uridine diphosphate tank 11, a UGT glycoside transferase tank 12, and a second sodium hydroxide solution tank 13. The outlet of the second reaction vessel 8 is connected via a pipe to a concentration tank 14. The lower outlet of the concentration tank 14 is connected via a pipe to a dryer 15. The outlet of the dryer 15 is connected to a glucosylstevioside tank 16. Rebaudioside A is used as the raw material, β-cyclodextrin or maltodextrin as excipients, and cyclodextrin glucoside transferase (…) is added… A glycosylation reaction was carried out using 3.0L of rebaudioside A. The pH was adjusted to 5.5-5.8 with sodium hydroxide to obtain a glucosylstevioside solution with rebaudioside A as the substrate. Then, the solution was hydrolyzed by a saccharifying enzyme to obtain low-glycosylated chain-length glucosylstevioside. This solution was then reacted with sucrose, UGT glycosyltransferase UGT76G4 (derived from Chinese patent CN115094074B), sucrose synthase (derived from sucrose synthase AtSuSy in Chinese patent CN119220516A), and uridine diphosphate glucose. The pH was adjusted to 7-8 to further convert unreacted rebaudioside A in the solution into rebaudioside I. After ethanol analysis, the solution was concentrated and dried to obtain glucosylstevioside. This process transformed the residual substrate into a steviol glycoside with a relatively better taste, improving the taste and quality of the product.

[0030] The outlet of the second reaction vessel 8 is connected to an adsorption resin column 17 via a pipeline. The inlet of the adsorption resin column 17 is connected to a first ethanol tank 18, a second ethanol tank 19, and a third ethanol tank 20 via pipelines. The outlet of the adsorption resin column 17 is connected to an eluent tank 21 via a pipeline. The outlet of the eluent tank 21 is connected to the concentration tank 14 via a pipeline. First, elution is performed using 10-15% ethanol in the first ethanol tank 18 to remove residual dextrin, monosaccharides, and proteins. Then, elution is performed using 20-30% ethanol in the second ethanol tank 19 to remove weakly adsorbed impurities such as highly glycosylated glycosides. Finally, elution is performed using 60-70% ethanol in the third ethanol tank 20 to obtain the target product, glucosylstevioside. This gradient elution improves the purity of the product and enhances its taste.

[0031] The inlet of the adsorption resin column 17 is connected to a purified water tank 22 via a pipeline. The purified water can be used to clean the adsorption resin column 17, removing residual impurities, ensuring stable adsorption performance, extending its service life, and reducing equipment maintenance costs. The outlet of the adsorption resin column 17 is connected to a water-to-alcohol tank 23 via a pipeline, and the outlet of the water-to-alcohol tank 23 is connected to the concentration tank 14 via a pipeline. The purified water can be used to top-wash the desorbed adsorption resin column 17, and then concentrated to obtain glucosylstevioside, thus improving the product yield.

[0032] The top outlet of the concentration tank 14 is connected to an ethanol recovery tank 24 via a pipeline, and the outlet of the ethanol recovery tank 24 is connected to the ethanol tank via a pipeline. This achieves ethanol recycling again, further reducing production costs and environmental pollution.

[0033] The outlet of the second reaction vessel 8 is connected to a plate and frame filter press 25 via a pipeline. The liquid phase outlet of the plate and frame filter press 25 is connected to the adsorption resin column 17 via a pipeline to remove insoluble substances such as proteins, obtaining a filtered clear liquid. The liquid phase outlet of the plate and frame filter press 25 is connected to an ultrafiltration membrane device 26 via a pipeline. The ultrafiltration membrane device 26 has a molecular weight cutoff of 3-10 kDa. The clear liquid outlet of the ultrafiltration membrane device 26 is connected to the adsorption resin column 17 via a pipeline, further removing impurities and improving the purity of the product.

[0034] The first reaction vessel 1 is equipped with a first online pH sensor 27, and the outlet of the first sodium hydroxide solution tank 5 is equipped with a first automatic shut-off valve 28. The first online pH sensor 27 and the first automatic shut-off valve 28 are interlocked to the control system. The second reaction vessel 8 is equipped with a second online pH sensor 29, and the outlet of the second sodium hydroxide solution tank 13 is equipped with a second automatic shut-off valve 30. The second online pH sensor 29 and the second automatic shut-off valve 30 are interlocked to the control system. The control system adjusts the opening and closing of the first automatic shut-off valve 28 and the second automatic shut-off valve 30 according to the values ​​of the first online pH sensor 27 and the second online pH sensor 29 to ensure that the pH value of the materials in the first reaction vessel 1 and the second reaction vessel 8 is at a stable value, which is conducive to the reaction.

[0035] The outlet of the raw material tank 2 is connected to a crushing tank 31 via a pipeline. The outlet of the crushing tank 31 is connected to a dryer 32 via a pipeline. The outlet of the dryer 32 is connected to the first reaction tank 1 via a pipeline. By crushing the raw material rebaudioside A into small particles with uniform particle size, the reaction efficiency and yield are improved.

[0036] It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the present invention. Furthermore, it should be understood that after reading the teachings of this invention, those skilled in the art can make various alterations or modifications to the invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. An apparatus for the stepwise preparation of glucosylsteviosides using multiple enzymes, characterized in that: The system includes a first reaction vessel, whose inlet is connected via pipes to a raw material vessel, a dextrin vessel, a cyclodextrin glucosyltransferase vessel, and a first sodium hydroxide solution vessel. The outlet of the first reaction vessel is connected via pipes to a hydrolysis vessel, whose inlet is connected via pipes to a saccharifying enzyme vessel. The outlet of the hydrolysis vessel is connected via pipes to a second reaction vessel, whose inlet is connected via pipes to a sucrose vessel, a sucrose synthase vessel, a uridine diphosphate vessel, a UGT glycosyltransferase vessel, and a second sodium hydroxide solution vessel. The outlet of the second reaction vessel is connected to a concentration tank via a pipeline, the lower outlet of the concentration tank is connected to a dryer via a pipeline, and the outlet of the dryer is connected to a glucosylstevioside tank.

2. The apparatus for stepwise preparation of glucosylsteviosides using multiple enzymes as described in claim 1, characterized in that: The outlet of the second reaction vessel is connected to an adsorption resin column via a pipeline. The inlet of the adsorption resin column is connected to a first ethanol tank, a second ethanol tank, and a third ethanol tank via pipelines. The outlet of the adsorption resin column is connected to an eluent tank via a pipeline. The outlet of the eluent tank is connected to a concentration tank via a pipeline.

3. The apparatus for stepwise preparation of glucosylsteviosides using multiple enzymes as described in claim 2, characterized in that: The inlet of the adsorption resin column is connected to a purified water tank via a pipe.

4. The apparatus for stepwise preparation of glucosylsteviosides using multiple enzymes as described in claim 3, characterized in that: The outlet of the adsorption resin column is connected to a water-top alcohol tank via a pipeline, and the outlet of the water-top alcohol tank is connected to the concentration tank via a pipeline.

5. The apparatus for stepwise preparation of glucosylsteviosides using multiple enzymes as described in claim 2, characterized in that: The top outlet of the concentration tank is connected to an ethanol recovery tank via a pipeline.

6. The apparatus for stepwise preparation of glucosylsteviosides using multiple enzymes as described in claim 2, characterized in that: The outlet of the second reaction vessel is connected to a plate and frame filter press via a pipeline, and the liquid phase outlet of the plate and frame filter press is connected to the adsorption resin column via a pipeline.

7. The apparatus for stepwise preparation of glucosylsteviosides using multiple enzymes as described in claim 6, characterized in that: The liquid phase outlet of the plate and frame filter press is connected to an ultrafiltration membrane device via a pipeline, and the clear liquid outlet of the ultrafiltration membrane device is connected to the adsorption resin column via a pipeline.

8. The apparatus for stepwise preparation of glucosylsteviosides using multiple enzymes as described in claim 1, characterized in that: The first reaction vessel is equipped with a first online pH sensor, and the outlet of the first sodium hydroxide solution vessel is equipped with a first automatic shut-off valve. The first online pH sensor and the first automatic shut-off valve are interlocked to the control system.

9. The apparatus for stepwise preparation of glucosylsteviosides using multiple enzymes as described in claim 1, characterized in that: The second reaction vessel is equipped with a second online pH sensor, and the outlet of the second sodium hydroxide solution vessel is equipped with a second automatic shut-off valve. The second online pH sensor and the second automatic shut-off valve are interlocked to the control system.

10. The apparatus for stepwise preparation of glucosylsteviosides using multiple enzymes as described in claim 1, characterized in that: The outlet of the raw material tank is connected to a crushing tank via a pipeline, the outlet of the crushing tank is connected to a dryer via a pipeline, and the outlet of the dryer is connected to the first reaction tank via a pipeline.

Images