A highly efficient method for purifying natamycin
By using a mixed solvent system under neutral conditions for natamycin purification, the problems of complex processes, high costs, and excessive wastewater in existing processes have been solved, achieving efficient and environmentally friendly natamycin purification with a purity of over 98% and a yield of over 95%.
Patent Information
- Application Number
- CN202211336621.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2022-10-28
- Publication Date
- 2026-01-20
- Estimated Expiration
- 2042-10-28
AI Technical Summary
Existing natamycin purification processes are complex, costly, generate large amounts of wastewater, and have low yields. Furthermore, natamycin is easily degraded under alkaline conditions, resulting in low purity and yield.
Natamycin was purified using a mixed solvent system under neutral conditions, including crude pulping, filtration and rinsing, and drying. The solubility difference and pH of the mixed solvent were utilized to control impurities, and rinsing was used to remove impurities, thus avoiding degradation under alkaline conditions.
It simplifies the purification process, reduces production costs, improves the purity and yield of natamycin, reduces wastewater volume, and is suitable for industrial production.
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Figure CN115925771B_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the field of food preservatives, in particular to a high-efficiency natamycin purification method. BACKGROUND
[0002] In 1955, Struyk et al. isolated Streptomyces natalensis from the soil of Natal state, South Africa, and first isolated natamycin from it. So far, natamycin is a new biological preservative synthesized by fermentation of Streptomyces natalensis, Streptomyces chattanoogensis, or Streptomyces ahygroscopicus, etc. and refined by biotechnology.
[0003] Natamycin is a food preservative with no odor, no taste, low dose and high safety, also known as natamycin, is a kind of active polyene macrolide antibiotic, which has high inhibitory capacity to various molds and yeasts. Natamycin is a white to cream white crystalline powder without odor and taste, which is usually in enolic structure. Its mechanism of action is to combine with fungal ergosterol and other sterol groups, block ergosterol biosynthesis, distort cell membrane, eventually lead to leakage, and cause cell death.
[0004] Natamycin is mainly used for food preservation to prevent mold and yeast caused by corruption. It has different antibacterial range from nisin and epsilon-polylysine, and can be used in combination, which is the most widely used biological preservative in the world.
[0005] At present, the purification of natamycin in industry is mainly through resin resolution and then crystallization or dissolution and then recrystallization.
[0006] For example, "Macroporous Resin Separation and Purification of Natamycin Process Research" by Wang Haiyan uses resin purification process, which has the disadvantages of complex process, long production cycle, high cost, large amount of wastewater discharge, etc. For example, "Natamycin Crude Powder Refining Process Optimization" by Li Congxin uses recrystallization combined with elution method to improve the purity of crude product. The solvent for redissolution is 75% methanol aqueous solution with a concentration of 25g / L, the dissolution pH is 13.0. The crystallization pH is 6.0-7.0, and the elution solvent for the finished product is 75% acetone aqueous solution. Natamycin is easily degraded under alkaline conditions, resulting in loss of yield. This process has many steps, large amount of wastewater, complex process and low yield. SUMMARY
[0007] The purpose of the present application is to solve the problems existing in the prior art and provide a high-efficiency natamycin purification method.
[0008] To achieve the above purpose, the present application is implemented according to the following technical scheme:
[0009] An efficient natamycin purification method, comprising the following steps:
[0010] S1, crude pulp beating: two kinds of dichloromethane, chloroform, acetone, butanone, methanol, ethanol, petroleum ether, n-hexane are mixed to obtain a mixed solution, the pH of the mixed solution is adjusted to 6.0-7.0, then the natamycin crude product is added to the mixed solution to form a slurry;
[0011] S2, filter leaching: the slurry of step S1 is filtered to obtain a natamycin wet product, and the natamycin wet product after filtering is leached with an organic solvent;
[0012] S3, drying: the natamycin wet product after leaching in step S2 is dried to obtain a natamycin purified product.
[0013] Further, the volume ratio of the two kinds of dichloromethane, chloroform, acetone, butanone, methanol, ethanol, petroleum ether, n-hexane is 1:2-1:6.
[0014] Further, the pH of the mixed solution in step S1 is adjusted using hydrochloric acid.
[0015] Preferably, the mass and volume ratio of the natamycin crude product to the mixed solution in step S1 is 1:10-30.
[0016] Preferably, the organic solvent in step S2 is one of dichloromethane, chloroform, acetone, butanone, methanol, ethanol, petroleum ether, n-hexane, and the volume fraction of the organic solvent is 65%-85%.
[0017] Preferably, the natamycin wet product after leaching in step S3 is dried using a drying device, which is one of an oven, a flash dryer, and a double-cone dryer.
[0018] Compared with the prior art, the present application purifies natamycin according to its solubility in different solvents and pH, first controls the purification under neutral conditions, avoids the degradation of natamycin when redissolved under alkaline conditions in the recrystallization process, improves the overall yield, and reduces the amount of wastewater; discards the scheme of high solubility and high impurity content of a single solvent in the traditional purification process, uses a mixed solvent for beating, realizes low solubility of natamycin and easy dissolution of impurities in natamycin, and selects a leaching agent in the natamycin extraction process to remove symbiotic similar impurities during the leaching process, further improving the purity of natamycin.
[0019] In summary, this invention optimizes the existing natamycin purification process, simplifies the purification method, reduces production costs, and improves production efficiency. The product purity can reach over 98%, and the product yield can reach over 95%. This invention requires less equipment investment, is easy to operate, has minimal environmental pollution, and is simple to operate, making it suitable for industrial production. Attached Figure Description
[0020] Figure 1 The liquid chromatogram of natamycin purified in Example 1 is shown.
[0021] Figure 2 The liquid chromatogram of natamycin purified in Example 2 is shown.
[0022] Figure 3 The liquid chromatogram of natamycin purified in Example 3 is shown.
[0023] Figure 4 The liquid chromatogram of natamycin purified in Example 4 is shown. Detailed Implementation
[0024] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. The specific embodiments described herein are for illustrative purposes only and are not intended to limit the invention.
[0025] Example 1
[0026] For comparison, purification was performed using recrystallization. 10 grams of crude natamycin with a purity of 92.3% was weighed using a balance to conduct a purification test.
[0027] 10 g of the crude product was placed in a 250 mL beaker and dissolved in 75% methanol solution. Hydrochloric acid was added dropwise to adjust the pH to 6.5-7.0 for recrystallization. The product was then washed with 30 mL of methanol to obtain the purified product. The purified product was dried in a vacuum drying oven at 60 °C to obtain 8.16 g of the final product. High-performance liquid chromatography (HPLC) analysis showed that the purity of the final product reached 98.5%.
[0028] The dry matter content of the finished product prepared in this batch was 98.3%, as shown in the following figures. Figure 1 As shown, by Figure 1 It can be known that:
[0029]
[0030] The yield of each step was then calculated based on the solution volume and the measured solution concentration. The yield of natamycin in this embodiment was determined to be 87.10% based on the ratio of crude product * purity / purified product * purity: 8.16 * 0.983 / 10 * 0.923.
[0031] Example 2
[0032] 10 grams of natamycin crude product with purity of 92.3% was weighed by a balance for purification test.
[0033] 10 grams of crude product was put into a 250 mL beaker, 95 mL of ethanol was added and stirred for 30 min, then 75 mL of acetone solution was added and stirred for 50 min. Filtration was performed by a Buchner funnel, then 30 mL of methanol was used for elution to obtain the purified product. The purified product was dried in a vacuum drying oven at 60°C to obtain the finished product of 9.25 grams. The finished product was detected by high performance liquid chromatography and the purity reached 98.1%.
[0034] The finished product prepared in this batch had a dry content of 98.1%, and the results are shown in Table 1, and it can be seen from Table 1 that: Figure 2 Figure 2
[0035]
[0036] Further, the yield of each step was calculated according to the solution volume and the measured solution concentration, and the yield of natamycin in this example was determined to be: 9.25*0.981 / 10*0.923=98.30% according to the crude product purity / purified product purity.
[0037] Example 3
[0038] 10 grams of natamycin crude product with purity of 93.3% was weighed by a balance for purification test.
[0039] 10 grams of crude product was put into a 250 mL beaker, 90 mL of methanol was added and stirred for 30 min, then 75 mL of acetone solution was added and stirred for 50 min. Filtration was performed by a Buchner funnel, then 30 mL of ethanol was used for elution to obtain the purified product. The purified product was dried in a vacuum drying oven at 60°C to obtain the finished product of 9.16 grams. The finished product was detected by high performance liquid chromatography and the purity reached 98.5%.
[0040] The finished product prepared in this batch had a dry content of 98.5%, and the results are shown in Table 2, and it can be seen from Table 2 that: Figure 3 Figure 3
[0041]
[0042] Further, the yield of each step was calculated according to the solution volume and the measured solution concentration, and the yield of natamycin in this example was determined to be: 9.11*0.985 / 10*0.923=97.80% according to the crude product purity / purified product purity.
[0043] Example 4
[0044] 10 grams of natamycin crude product with purity of 94.5% was weighed by a balance for purification test.
[0045] Put 10 grams of the crude product into a 250 mL beaker, add 85 mL of methanol and stir for 30 min, then add 75 mL of acetone solution and stir for 50 min. Filter with a Buchner funnel, then rinse with 30 mL of methanol to obtain the purified product. Dry the purified product in a vacuum drying oven at 60°C to obtain the finished product 9.10 grams. The purity of the finished product is 98.9% as determined by high performance liquid chromatography.
[0046] The content of the finished product prepared in this batch is 98.9%, and the results are shown in Table 1. Figure 4 As can be seen from Table 1: Figure 4 As can be seen from Table 1:
[0047]
[0048] Further, according to the volume of the solution and the determined concentration of the solution, the yield of each step is calculated, and according to the purity of the crude product / purity of the purified product, the yield of natamycin in this embodiment is determined to be: 9.10*0.989 / 10*0.923=97.50%.
[0049] The technical solution of the present application is not limited to the above specific embodiments, and any technical modification made according to the technical solution of the present application falls within the protection scope of the present application.
Claims
1. A highly efficient method for purifying natamycin, characterized in that, Includes the following steps: S1. Crude product pulping: Take two of the following: dichloromethane, chloroform, acetone, butanone, methanol, ethanol, petroleum ether, and n-hexane to obtain a mixed solution. Adjust the pH of the mixed solution to 6.0-7.0, and then add the crude natamycin to the mixed solution and stir to form a slurry. S2, filtration and rinsing: The slurry from step S1 is filtered to obtain wet natamycin. The filtered wet natamycin is then rinsed with an organic solvent. S3. Drying: Dry the wet natamycin product after rinsing in step S2 to obtain the purified natamycin product.
2. The efficient natamycin purification method according to claim 1, characterized in that: The volume ratio of two of the following: dichloromethane, chloroform, acetone, butanone, methanol, ethanol, petroleum ether, and n-hexane is 1:2 to 1:
6.
3. The efficient natamycin purification method according to claim 1, characterized in that: In step S1, hydrochloric acid is used to adjust the pH of the mixed solution.
4. The efficient natamycin purification method according to claim 1, characterized in that: In step S1, the mass-to-volume ratio of crude natamycin to the mixed solution is 1:10-30.
5. The efficient natamycin purification method according to claim 1, characterized in that: The organic solvent in step S2 is one of dichloromethane, chloroform, acetone, butanone, methanol, ethanol, petroleum ether, and n-hexane, and the volume fraction of the organic solvent is 65%-85%.
6. The efficient natamycin purification method according to claim 1, characterized in that: In step S3, the rinsed natamycin wet product is dried using a drying device, which is one of an oven, a flash dryer, or a double cone dryer.