Method for recovering mother liquor of ergothioneine

Abstract

The invention relates to the technical field of biochemical engineering, in particular to an ergothioneine mother liquor recovery method. The method comprises the following steps: quantitatively passing diluted mother liquor through cationic resin, adsorbing the ergothioneine on the resin, washing the resin with water to remove impurities, then carrying out top washing on the ergothioneine on the resin by using a certain amount of methionine solution, collecting a top washing solution containing the ergothioneine, and concentrating the top washing solution to a certain proportion to obtain the ergothioneine. The concentrated solution passes through chromatographic separation resin to remove methionine, a qualified material containing ergothioneine is collected, and the collected qualified material is concentrated, subjected to alcohol precipitation and crystallized to obtain a recovered ergothioneine product. According to the method, the ergothioneine in the mother liquor can be effectively recycled, the technological process is simple, the treatment cost is low, the extraction yield is increased, the mother liquor recycling rate can reach 61.08%, the product purity can reach 98.63%, the treatment effect is good, and the method is suitable for large-scale industrialized production.

Description

Technical Field

[0001] The invention relates to the technical field of biochemical engineering, and in particular to a method for recovering a mother liquor of ergothioneine. Background Art

[0002] L-Ergothioneine (EGT) is a rare natural chiral amino acid antioxidant and the only natural 2-thioimidazole amino acid (2-thio-imidazole) discovered so far. It mainly exists in the form of thioketone under physiological pH conditions. The stability of the thiocarbonyl group is higher than that of the sulfhydryl group. Therefore, EGT will not spontaneously oxidize and has good pH stability and thermal stability.

[0003] Ergothioneine has the functions of scavenging free radicals, detoxifying, maintaining DNA growth, anti-radiation, and anti-aging. It is a unique cell physiological protector. Secondly, the presence of the transporter ETT makes EGT the only antioxidant that can be transported between cells, which can protect the nervous system, inhibit developmental defects, and protect the liver. In view of the physiological activity of EGT, it has broad application prospects in food, cosmetics, medicine and other fields.

[0004] The existing thioneine mother liquor recovery means are complex, have low universality, small single processing volume, high cost, involve many equipment and long process time; such as the Chinese invention patent "CN114262297A a method for preparing thioneine", with boiling combined with freezing plus film passing, reduced pressure evaporation treatment, resin treatment, etc., the equipment is multi-complicated operation; the Chinese invention patent "CN113666873A a method for industrial preparation of high-purity thioneine" uses resin columns for ion exchange many times, which takes a long time, has a large amount of wastewater and is costly; therefore, a more effective and practical thioneine mother liquor recovery method is an existing problem that needs to be solved. Summary of the invention

[0005] The object of the present invention is to provide a mother liquor recovery method of thioneine, to solve the problems that existing thioneine mother liquor recovery means are complicated, universality is low, single processing capacity is small, cost is high, involves many equipments and process is time-consuming.

[0006] To achieve the above object, the present invention adopts the following technical scheme: a mother liquor recovery method of thioneine, comprising the following steps:

[0007] S1, pass thioneine crystallization mother liquor through cation exchange resin, thioneine is adsorbed on the cation exchange resin;

[0008] S2, with methionine solution, the thioneine top wash on the cation exchange resin is washed down, collects top wash liquid, concentrates, obtains concentrated solution;

[0009] S3, concentrated solution is passed through chromatographic separation resin to remove methionine, and qualified materials containing thioneine are collected;

[0010] S4, the collected qualified materials are concentrated, alcohol precipitated and crystallized to obtain thioneine recovery products.

[0011] Further, in the S1, the content of ergothioneine in the crystallization mother liquor is 20-30 g / L.

[0012] Furthermore, in the S1, the cation exchange resin is a strongly acidic cation exchange resin, and the model is 001*16.

[0013] In the S1, the amount of pure ergothioneine in the crystallization mother liquor corresponding to each liter of cation exchange resin is 50-60 g.

[0014] Furthermore, in the S2, the content of methionine in the methionine solution is 30-40 g / L.

[0015] Furthermore, in the S1-S2, the molar ratio of methionine to ergothioneine is 1.2 to 1.3:1.

[0016] Furthermore, in the S2, the specific gravity of the concentrated liquid is 1.1 to 1.2.

[0017] Furthermore, in the S3, the chromatographic separation resin is a sodium type, the chromatographic separation resin feed amount is 40 to 50 g of concentrated liquid per liter of chromatographic separation resin, the chromatographic separation temperature is 30 to 60° C., the flow rate is 20 to 25 mL / min, and deionized water is used for desorption.

[0018] Furthermore, in the S3, the methionine content in the qualified material is less than 0.5 g / L.

[0019] Further, in described S4, the purity of the obtained thioneine recovered product is more than 98%, and the recovery rate of the mother liquor can reach more than 60%.

[0020] Beneficial effects of the present invention:

[0021] 1. The present invention adopts 001*16 type positive resin to adsorb thioneine in the mother liquor to effectively remove the impurities in the mother liquor, and adopts methionine to elute the thioneine on the positive resin to have a better effect, and does not introduce other impurities;

[0022] 2. The present invention adopts sodium type chromatographic separation resin to effectively remove methionine in the eluent, improve the purity of ergothioneine in the chromatographic qualified material, and effectively recover ergothioneine in the mother liquor;

[0023] 3. The present invention has good recovery effect, high universality, large single processing volume, high purity, and only needs simple subsequent processing of the recovered products to meet quality requirements, which is suitable for large-scale factory production. DETAILED DESCRIPTION

[0024] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention.

[0025] Example 1

[0026] Take 2.5L of the diluted mother liquor with a thioneine content of 20g / L, and pass it through 1 liter of 001*16 type cationic resin at a flow rate of 2BV / h. After the mother liquor is completed, rinse the resin with 2 times the resin volume of deionized water until the impurities are completely washed out.

[0027] The adsorbed resin was fed with 35 g / L methionine solution to elute ergothioneine, the molar ratio of methionine to ergothioneine being 1.25:1. After the methionine solution was fed, the resin was rinsed with deionized water twice the volume of the resin, and the eluate was collected and concentrated to a specific gravity of 1.15.

[0028] The concentrate was purified by using 2 liters of sodium-type chromatographic separation resin, with 90 g of concentrate fed each time. Qualified materials were collected based on the liquid phase results, and the methionine content in the qualified materials was controlled to be less than 0.5 g / L.

[0029] The collected qualified material was concentrated and crystallized by alcohol precipitation to obtain 30.6 g of pure ergothioneine, with a purity of 98.63% and a mother liquor recovery yield of 60.36%.

[0030] Example 2

[0031] Take 2.5L of the diluted mother liquor with a thioneine content of 20g / L, and pass it through 1 liter of 001*16 type cationic resin at a flow rate of 2V / h. After the mother liquor is completed, rinse the resin with deionized water twice the volume of the resin until the impurities are completely washed out.

[0032] The adsorbed resin was fed with 30 g / L methionine solution to elute ergothioneine, the molar ratio of methionine to ergothioneine being 1.25:1. After the methionine solution was fed, the resin was rinsed with deionized water twice the volume of the resin, and the eluate was collected and concentrated to a specific gravity of 1.15.

[0033] The concentrate was purified by using 2 liters of sodium-type chromatographic separation resin, with 80 g of concentrate fed each time. Qualified materials were collected based on the liquid phase results, and the methionine content in the qualified materials was controlled to be less than 0.5 g / L.

[0034] The collected qualified material was concentrated and crystallized by alcohol precipitation to obtain 31 g of recovered ergothioneine, with a purity of 98.52% and a mother liquor recovery yield of 61.08%.

[0035] Example 3

[0036] Take 2L of the diluted mother liquor with a content of 30g / L of thioneine, and pass it through 1 liter of 001*16 type cationic resin at a flow rate of 2V / h. After the mother liquor is completed, rinse the resin with deionized water twice the volume of the resin until the impurities are completely washed out.

[0037] The adsorbed resin was fed with 40 g / L methionine solution to elute ergothioneine, the molar ratio of methionine to ergothioneine was 1.25:1, and after the methionine solution was fed, the resin was rinsed with deionized water twice the volume of the resin, and the eluate was collected and concentrated to a specific gravity of 1.15.

[0038] The concentrate was purified by using 2 liters of sodium-type chromatographic separation resin, with 80 g of concentrate fed each time. Qualified materials were collected based on the liquid phase results, and the methionine content in the qualified materials was controlled to be less than 0.5 g / L.

[0039] The collected qualified material was concentrated and crystallized to obtain 36.6g of recovered ergothioneine, with a purity of 98.31% and a mother liquor recovery yield of 59.97%.

[0040] In the above-mentioned embodiment of the present invention, ergothioneine and methionine are detected by high performance liquid chromatography (HPLC).

[0041] The detection conditions of ergothioneine are as follows: the chromatographic column is a Yuexu LP-C18 chromatographic column, the volume ratio of the mobile phase components is acetonitrile: pure water = 30:70, an ultraviolet detector is used, the detection wavelength is 254nm, the column temperature is 35°C, the injection volume is 10μL, and the flow rate is 1.0mL / min.

[0042] The detection conditions of methionine are:

[0043] Mobile phase A: Take 0.82g of anhydrous sodium acetate and 3.8g of sodium tetraborate decahydrate, dissolve them in 1L of water, stir thoroughly to completely dissolve the borate crystals, adjust the pH to 8.2 with hydrochloric acid, filter with a 0.22um water filter membrane, put it in an ultrasonic cleaner for 30 minutes, and set aside.

[0044] Mobile phase B: Mix 450 mL of chromatographic-grade methanol, 450 mL of acetonitrile, and 100 mL of water, filter with a 0.22 um organic filter membrane, and place in an ultrasonic cleaner for 60 min for later use.

[0045] The chromatographic conditions were as follows:

[0046] Chromatographic column: YMC-Triart C18 (4.6*250mm, 5μm);

[0047] Flow rate: 1.5 mL / min;

[0048] Detection wavelength: 338nm;

[0049] Column temperature: 40℃.

[0050] The gradient elution program was:

[0051]

[0052] The present invention is not limited to the above-mentioned optimal implementation mode. Anyone can derive other various forms of products under the inspiration of the present invention. However, no matter what changes are made in the shape or structure, all technical solutions that are the same or similar to those of the present application fall within the protection scope of the present invention.

Claims

1. a mother liquor recovery method for thioneine, characterized in that, The following steps are involved: S1, pass thioneine crystallization mother liquor through cation exchange resin, thioneine is adsorbed on the cation exchange resin; S2, with methionine solution, the thioneine top wash on the cation exchange resin is washed down, collects top wash liquid, concentrates, obtains concentrated solution; S3, concentrated solution is passed through chromatographic separation resin to remove methionine, and qualified materials containing thioneine are collected; S4, the collected qualified materials are concentrated, alcohol precipitated and crystallized to obtain thioneine recovery products.

2. a mother liquor recovery method of thioneine according to claim 1, is characterized in that: In the described S1, the content of ergothioneine in the crystallization mother liquor is 20-30 g / L.

3. a mother liquor recovery method of thioneine according to claim 1, is characterized in that: In the S1, the cation exchange resin is a strongly acidic cation exchange resin, and the model is 001*16.

4. a mother liquor recovery method of thioneine according to claim 1, is characterized in that: In the S1, the amount of pure ergothioneine in the crystallization mother liquor corresponding to each liter of cation exchange resin is 50-60 g.

5. a mother liquor recovery method of thioneine according to claim 1, is characterized in that: In the S2, the content of methionine in the methionine solution is 30-40 g / L.

6. a mother liquor recovery method of thioneine according to claim 1, is characterized in that: In the S1-S2, the molar ratio of methionine to ergothioneine is 1.2-1.3:

1.

7. a mother liquor recovery method of thioneine according to claim 1, is characterized in that: In the S2, the specific gravity of the concentrated liquid is 1.1 to 1.

2.

8. a mother liquor recovery method of thioneine according to claim 1, is characterized in that: In the S3, the chromatographic separation resin is a sodium type, the chromatographic separation resin feed amount is 40 to 50 g of concentrated solution per liter of chromatographic separation resin, the chromatographic separation temperature is 30 to 60° C., the flow rate is 20 to 25 mL / min, and deionized water is used for desorption.

9. a mother liquor recovery method of thioneine according to claim 1, is characterized in that: In the S3, the methionine content in the qualified material is less than 0.5 g / L.

10. according to the mother liquor recovery method of a kind of thioneine described in claim 1-9 any one, it is characterized in that: In the described S4, the purity of the obtained thioneine recovered product is more than 98%, and the recovery rate of the mother liquor can reach more than 60%.

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