Extraction device of mogroside V
By optimizing the extraction device for mogroside V and utilizing glycosyltransferase fermentation and various separation technologies, the problems of low yield and high cost in the extraction process of mogroside V were solved, achieving efficient and low-cost extraction results.
Patent Information
- Application Number
- CN202423236048.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-26
- Publication Date
- 2025-12-30
- Estimated Expiration
- 2034-12-26
AI Technical Summary
Existing methods for extracting mogroside V are subject to the long fruit ripening cycle and significant impact on quality, resulting in low yields, high costs, and a large amount of manpower and resources required for separation and purification.
The fermentation broth of Escherichia coli containing glycosyltransferases UGT-MS1 and UGT-MS2 was reacted with mogroside IIE, uridine diphosphate glucose and calcium salt. The mixture was eluted and impurities removed by macroporous resin column and anion exchange resin column. Calcium chloride and diatomaceous earth were used as flocculants, and ultrafiltration membrane device was used for filtration to optimize the fermentation and separation process.
It improved the yield and purity of mogroside V, shortened the production cycle, reduced production costs, and improved separation efficiency and product purity through flocculants and ultrafiltration membranes.
Smart Images
Figure CN223732785U_ABST
Abstract
Description
Technical Field
[0001] This utility model relates to the field of biotechnology, specifically to an extraction device for mogroside V. Background Technology
[0002] Mogrosides are the main terpenoid active substances in monk fruit, belonging to the cucurbitane-type tetracyclic triterpenoid saponins. They are also the main sweeteners in monk fruit, with a sweetness approximately 300 times that of sucrose. In mature fruit, mogroside V is the main sweetener, while a small amount of mogroside IV is also present. This sweet mogroside is widely used in the market and is called mogroside sweetener. In immature fruit, mogrosides II and III are the main components, and at this stage, the fruit has no sweetness and may even have a strong bitter taste.
[0003] Currently, the main method for obtaining mogrosides is plant extraction. However, this method is significantly affected by the season and the quality of the mogrosides. Furthermore, the content of mogrosides varies greatly due to differences in the quality of the mogrosides, and the components also differ considerably. This results in a large consumption of manpower and resources during the separation and purification process, and the method is limited by the different separation methods available. Most importantly, the long ripening cycle of mogrosides means that the annual yield is far from meeting the growing demand, leading to high production costs and severely restricting its output. Summary of the Invention
[0004] The technical problem to be solved by this utility model is to provide an extraction device for mogroside V that has a high product yield and low production cost, 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 extraction apparatus for mogroside V includes a reaction tank, the inlet of which is connected to a fermentation broth tank, a mogroside IIE tank, a uridine diphosphate glucose tank and a salt tank via pipes, and the outlet of which is connected to a conversion broth tank via pipes.
[0007] The outlet of the conversion liquid tank is connected to a macroporous resin column via a pipe. The inlet of the macroporous resin column is connected to an eluent tank via a pipe. The outlet of the macroporous resin column is connected to an eluent tank. The outlet of the eluent tank is connected to an anion exchange resin column via a pipe. The outlet of the anion exchange resin column is connected to a concentration tank. The outlet of the concentration tank is connected to a dryer. The outlet of the dryer is connected to a mogroside V tank.
[0008] As an improved technical solution, the outlet of the fermentation broth tank is connected to a first centrifuge via a pipeline, the solid phase outlet of the first centrifuge is connected to a resuspension tank, the inlet of the resuspension tank is connected to a phosphate buffer solution tank via a pipeline, the outlet of the resuspension tank is connected to a homogenizer via a pipeline, the outlet of the homogenizer is connected to a second centrifuge via a pipeline, the liquid phase outlet of the second centrifuge is connected to an enzyme clear liquid tank, and the outlet of the enzyme clear liquid tank is connected to the reaction tank via a pipeline.
[0009] As an improved technical solution, the outlet of the conversion liquid tank is connected to an insulated tank via a pipeline, and the outlet of the insulated tank is connected to the macroporous resin column via a pipeline.
[0010] As an improved technical solution, the inlet of the heat preservation tank is connected to a calcium chloride tank and a diatomaceous earth tank through pipes, the outlet of the heat preservation tank is connected to a filter, the liquid phase outlet of the filter is connected to a filtrate tank, and the outlet of the filtrate tank is connected to the macroporous resin column through a pipe.
[0011] As an improved technical solution, the outlet of the filtrate tank is connected to an ultrafiltration membrane device via a pipeline, and the outlet of the ultrafiltration membrane device is connected to the macroporous resin column.
[0012] As an improved technical solution, the outlet of the ultrafiltration membrane device is connected to an regulating tank via a pipeline, and the outlet of the regulating tank is connected to the macroporous resin column.
[0013] As a preferred technical solution, the inlet of the macroporous resin column is connected to a deionized water tank and an ethanol solution tank via pipes.
[0014] Due to the adoption of the above technical solution, the beneficial effects of this utility model are:
[0015] This invention relates to an extraction device for mogroside V, comprising a reaction vessel. The inlet of the reaction vessel is connected via pipes to a fermentation broth tank, a mogroside IIE tank, a uridine diphosphate glucose tank, and a salt tank. The outlet of the reaction vessel is connected via pipes to a conversion broth tank. The outlet of the conversion broth tank is connected via pipes to a macroporous resin column. The inlet of the macroporous resin column is connected via pipes to an eluent tank. The outlet of the macroporous resin column is connected via pipes to an eluent tank. The outlet of the eluent tank is connected via pipes to an anion exchange resin column. The outlet of the anion exchange resin column is connected to a concentration tank. The outlet of the concentration tank is connected to a dryer. The outlet of the dryer is connected to a mogroside V tank. By culturing Escherichia coli containing glycosyltransferases UGT-MS1 and UGT-MS2 respectively, the culture broth is fermented to obtain a fermentation broth. The fermentation broth is then reacted with one or more of the following: mogroside IIE, uridine diphosphate glucose, and calcium and magnesium salts. The resulting conversion solution is eluted onto a macroporous resin column. The eluent is then passed through an anion exchange resin column for impurity removal. Finally, the effluent is concentrated and dried to obtain mogroside V. This process improves the product yield and purity. Furthermore, after passing through the anion exchange resin column, mogroside V changes from its original pale yellow or brownish-yellow color to a white product. Moreover, the eluent from the macroporous resin column does not require ethanol concentration and recovery and can be directly fed onto the column, shortening the production steps and cycle.
[0016] The specific fermentation process is as follows: the culture medium is inoculated with the bacterial culture at an inoculation rate of 5-10% v / v. The initial pH of the fermentation medium is 7.0, and the initial air flow rate is 0.5 m³ / h. 3 The fermentation process was carried out at a concentration of 0.05 MPa, a pressure of 35–38°C, and a rotation speed of 250–300 rpm. During fermentation, the stirring speed was automatically coupled to the dissolved oxygen level. The temperature was automatically controlled by the fermenter. The pH was maintained by adding 50% ammonia solution. The dissolved oxygen (DO) value was kept at 20–30% for the first 24 hours of fermentation. The culture was continued until the OD value of the bacterial culture reached a certain level. 600 The pH value was 30–60. IPTG was added to induce fermentation at a final concentration of 0.4% wt, and the mixture was fermented at 30°C for 24 hours to obtain the fermentation broth. The nucleotide sequence of UGT-MS1 is shown in SEQ ID NO.1, and the nucleotide sequence of UGT-MS2 is shown in SEQ ID NO.2.
[0017] UGT-MS1 nucleotide sequence (SEQ ID NO.1)
[0018]
[0019] UGT-MS2 nucleotide sequence (SEQ ID NO.2)
[0020]
[0021] The fermentation broth tank of this invention has its outlet connected to a first centrifuge via a pipeline. The solid phase outlet of the first centrifuge is connected to a resuspension tank, the inlet of which is connected to a phosphate buffer solution tank via a pipeline. The outlet of the resuspension tank is connected to a homogenizer via a pipeline, and the outlet of the homogenizer is connected to a second centrifuge via a pipeline. The liquid phase outlet of the second centrifuge is connected to an enzyme clear liquid tank, and the outlet of the enzyme clear liquid tank is connected to the reaction tank via a pipeline. After the fermentation broth enters the first centrifuge, the bacterial cells are collected. Then, the bacterial cells are resuspended in a phosphate buffer solution. The resulting resuspension is then homogenized, and finally separated by the second centrifuge to obtain the enzyme clear liquid. The resuspension and homogenization processes improve the dispersibility of the enzyme clear liquid, resulting in better enzyme catalytic effect on mogroside IIIE and increasing the product yield.
[0022] The outlet of the conversion liquid tank is connected to an insulated tank via a pipeline, and the outlet of the insulated tank is connected to the macroporous resin column via a pipeline. By maintaining the temperature at 80-90℃ for 20-40 minutes, the product yield is further improved.
[0023] The inlet of the insulated tank is connected to a calcium chloride tank and a diatomaceous earth tank via pipes. The outlet of the insulated tank is connected to a filter, and the liquid phase outlet of the filter is connected to a filtrate tank. The outlet of the filtrate tank is connected to the macroporous resin column via a pipe. Using calcium chloride and diatomaceous earth as flocculants is relatively inexpensive. These flocculants react with phosphates present in the conversion liquid to form calcium phosphate precipitates, which act as filter aids during filtration, accelerating the filtration speed of the filtrate and achieving phosphorus removal, thus reducing subsequent wastewater treatment costs.
[0024] The outlet of the filtrate tank is connected to an ultrafiltration membrane device via a pipeline, and the outlet of the ultrafiltration membrane device is connected to the macroporous resin column. Passing the obtained filtrate through the ultrafiltration membrane device can remove large protein molecules, thereby increasing the injection volume and separation efficiency of the resin in the later stages.
[0025] The outlet of the ultrafiltration membrane device is connected to an adjustment tank via a pipeline, and the outlet of the adjustment tank is connected to the macroporous resin column. By adjusting the pH of the obtained filtrate to 7.0-8.0 before loading it onto the column, subsequent elution is facilitated, thus improving the product yield.
[0026] The inlet of the macroporous resin column is connected to a deionized water tank and an ethanol solution tank via pipes. Washing with deionized water and ethanol improves the purity of the product. Attached Figure Description
[0027] The present invention will be further described below with reference to the accompanying drawings and embodiments.
[0028] Figure 1This is a structural schematic diagram of an embodiment of the present utility model;
[0029] The components include: 1. Reaction tank; 2. Fermentation broth tank; 3. Mogroside IIIE tank; 4. Uridine diphosphate glucose tank; 5. Salt tank; 6. Conversion broth tank; 7. Macroporous resin column; 8. Eluent tank; 9. Eluent tank; 10. Anion exchange resin column; 11. Concentrator; 12. Dryer; 13. Mogroside V tank; 14. First centrifuge; 15. Resuspension tank; 16. Phosphate buffer solution tank; 17. Homogenizer; 18. Second centrifuge; 19. Enzyme clearing solution tank; 20. Insulation tank; 21. Calcium chloride tank; 22. Diatomaceous earth tank; 23. Filter; 24. Filtrate tank; 25. Ultrafiltration membrane device; 26. Adjustment tank; 27. Deionized water tank; 28. Ethanol solution tank. Detailed Implementation
[0030] The present invention will be further described below with reference to the accompanying drawings and embodiments.
[0031] like Figure 1 As shown, an extraction apparatus for mogroside V includes a reaction tank 1. The inlet of the reaction tank 1 is connected via pipes to a fermentation broth tank 2, a mogroside IIIE tank 3, a uridine diphosphate glucose tank 4, and a salt tank 5. The outlet of the reaction tank 1 is connected via pipes to a conversion broth tank 6. The outlet of the conversion broth tank 6 is connected via pipes to a macroporous resin column 7. The inlet of the macroporous resin column 7 is connected via pipes to an eluent tank 8. The outlet of the macroporous resin column 7 is connected via pipes to an eluent tank 9. The outlet of the eluent tank 9 is connected via pipes to an anion exchange resin column 10. The outlet of the anion exchange resin column 10 is connected to a concentration tank 11. The outlet of the concentration tank 11 is connected to a dryer 12. The outlet of the dryer 12 is connected to a mogroside V tank 13. By culturing Escherichia coli containing glycosyltransferases UGT-MS1 and UGT-MS2 respectively, the resulting culture broth is fermented to obtain a fermentation broth. The fermentation broth is then reacted with one or more of the following: mogroside IIE, uridine diphosphate glucose, and calcium and magnesium salts, to obtain a conversion solution. This solution is then eluted onto a macroporous resin column 7. The eluent is then passed through an anion exchange resin column 10 for impurity removal. Finally, the effluent is concentrated and dried to obtain mogroside V as the final product. This process improves the product yield and purity. Furthermore, after passing through the anion exchange resin column 10, mogroside V changes from its original pale yellow or brownish-yellow color to a white product. Moreover, the eluent from the macroporous resin column 7 does not require ethanol concentration and recovery and can be directly fed onto the column, shortening the production steps and cycle.
[0032] The specific fermentation process is as follows: the culture medium is inoculated with the bacterial culture at an inoculation rate of 5-10% v / v. The initial pH of the fermentation medium is 7.0, and the initial air flow rate is 0.5 m³ / h. 3The fermentation process was carried out at a pressure of 0.05 MPa, a temperature of 35–38°C, and a rotation speed of 250–300 rpm. During fermentation, the stirring speed was automatically coupled to the dissolved oxygen level. The temperature was automatically controlled by the fermenter. The pH was maintained by adding 50% ammonia water. The dissolved oxygen (DO) value was kept at 20–30% before 24 hours of fermentation. When the OD600 value of the bacterial culture reached 30–60, IPTG was added at a final concentration of 0.4% wt to induce fermentation. The culture was then kept at a constant temperature of 30°C for 24 hours to obtain the fermentation broth.
[0033] The outlet of fermentation broth tank 2 is connected to a first centrifuge 14 via a pipeline. The solid phase outlet of the first centrifuge 14 is connected to a resuspension tank 15. The inlet of the resuspension tank 15 is connected to a phosphate buffer solution tank 16 via a pipeline. The outlet of the resuspension tank 15 is connected to a homogenizer 17 via a pipeline. The outlet of the homogenizer 17 is connected to a second centrifuge 18 via a pipeline. The liquid phase outlet of the second centrifuge 18 is connected to an enzyme clear liquid tank 19. The outlet of the enzyme clear liquid tank 19 is connected to the reaction tank 1 via a pipeline. After the fermentation broth enters the first centrifuge 14, the bacterial cells are collected. Then, the bacterial cells are resuspended in a phosphate buffer solution. The resulting resuspension is then homogenized. Finally, the enzyme clear liquid is obtained by separation in the second centrifuge 18. After resuspension and homogenization, the dispersibility of the enzyme clear liquid is improved, resulting in better enzyme catalytic effect on mogroside IIIE and increasing the product yield.
[0034] The outlet of the conversion liquid tank 6 is connected to an insulated tank 20 via a pipe, and the outlet of the insulated tank 20 is connected to the macroporous resin column 7 via a pipe. By maintaining the temperature at 80-90℃ for 20-40 minutes, the product yield is further improved.
[0035] The inlet of the insulated tank 20 is connected to a calcium chloride tank 21 and a diatomaceous earth tank 22 via pipes. The outlet of the insulated tank 20 is connected to a filter 23. The liquid phase outlet of the filter 23 is connected to a filtrate tank 24. The outlet of the filtrate tank 24 is connected to the macroporous resin column 7 via a pipe. Using calcium chloride and diatomaceous earth tank 22 as flocculants is relatively inexpensive. These flocculants react with the phosphates present in the conversion liquid to form calcium phosphate precipitate, which acts as a filter aid during filtration, accelerating the filtration speed of the filtrate and achieving phosphorus removal, thus reducing subsequent wastewater treatment costs.
[0036] The outlet of the filtrate tank 24 is connected to an ultrafiltration membrane device 25 via a pipeline, and the outlet of the ultrafiltration membrane device 25 is connected to the macroporous resin column 7. Passing the obtained filtrate through the ultrafiltration membrane device 25 can remove large protein molecules and improve the injection volume and separation efficiency of the subsequent resin.
[0037] The outlet of the ultrafiltration membrane device 25 is connected to an adjusting tank 26 via a pipe, and the outlet of the adjusting tank 26 is connected to the macroporous resin column 7. By adjusting the pH of the obtained filtrate to 7.0-8.0 before loading it onto the column, subsequent elution is facilitated, thus improving the product yield.
[0038] The inlet of the macroporous resin column 7 is connected to a deionized water tank 27 and an ethanol solution tank 28 via pipes. Washing with deionized water and ethanol improves the purity of the product.
[0039] 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. A device for extracting mogroside V, characterized in that: The reaction tank is communicated with the fermentation liquid tank, the mogroside IIE tank, the uridine diphosphate glucose tank and the salt tank through pipes respectively, and the outlet of the reaction tank is communicated with the conversion liquid tank through a pipe; The outlet of the conversion liquid tank is communicated with the macroporous resin column through a pipe, the inlet of the macroporous resin column is communicated with the eluent tank through a pipe, the outlet of the macroporous resin column is communicated with the eluate tank, the outlet of the eluate tank is communicated with the anion exchange resin column through a pipe, the outlet of the anion exchange resin column is communicated with the concentration tank, the outlet of the concentration tank is communicated with the dryer, and the outlet of the dryer is communicated with the mogroside V tank. 2.The mogroside V extraction device of claim 1, wherein: The outlet of the fermentation liquid tank is communicated with the first centrifuge through a pipe, the solid phase outlet of the first centrifuge is communicated with the resuspension tank, the inlet of the resuspension tank is communicated with the phosphate buffer solution tank through a pipe, the outlet of the resuspension tank is communicated with the homogenizer through a pipe, the outlet of the homogenizer is communicated with the second centrifuge through a pipe, the liquid phase outlet of the second centrifuge is communicated with the enzyme supernatant tank, and the outlet of the enzyme supernatant tank is communicated with the reaction tank through a pipe. 3.The mogroside V extraction device of claim 1, wherein: The outlet of the conversion liquid tank is communicated with the heat preservation tank through a pipe, and the outlet of the heat preservation tank is communicated with the macroporous resin column through a pipe. 4.The mogroside V extraction device of claim 3, characterized in that: The inlet of the heat preservation tank is respectively communicated with the calcium chloride tank and the diatomite tank through pipes, the outlet of the heat preservation tank is communicated with the filter, the liquid phase outlet of the filter is communicated with the filtrate tank, and the outlet of the filtrate tank is communicated with the macroporous resin column through a pipe. 5.The mogroside V extraction device of claim 4, characterized in that: The outlet of the filtrate tank is communicated with the ultrafiltration membrane device, and the outlet of the ultrafiltration membrane device is communicated with the macroporous resin column. 6.The mogroside V extraction device of claim 5, characterized in that: The outlet of the ultrafiltration membrane device is communicated with the adjustment tank, and the outlet of the adjustment tank is communicated with the macroporous resin column. 7.The mogroside V extraction device of claim 1, wherein: The inlet of the macroporous resin column is respectively communicated with the deionized water tank and the ethanol solution tank through pipes.