A kind of extraction process of vitamin K2

Abstract

The present invention proposes a vitamin K2 extraction process, comprising the following steps: (1) adding a filter aid to a vitamin K2 fermentation broth, stirring evenly, and then performing solid-liquid separation to obtain a fungus residue rich in vitamin K2; (2) drying and crushing the fungus residue to obtain fungus powder; (3) loading the fungus powder into a chromatography column, and compacting the fungus powder with clean gas; (4) adding an organic solvent for extraction from a feed port of the chromatography column at a set flow rate, collecting the effluent of the chromatography column, obtaining a vitamin K2 high-concentration extract, and after collecting 0.4-0.6 times the volume of the chromatography column, stopping the addition of the organic solvent, and pressing the liquid in the chromatography column with clean gas; (5) concentrating the vitamin K2 high-concentration extract to obtain a vitamin K2 ointment. Compared with the traditional stirring extraction process, the process of the present invention has the advantages of less use of extraction solvent, high extraction yield, and high content of vitamin K2 ointment, and is very valuable for industrialization.

Description

Technical Field

[0001] The invention relates to the technical field of vitamin extraction, and in particular to an extraction process of vitamin K2. Background Art

[0002] Vitamin K2, also known as menadione, is a fat-soluble vitamin that plays an important role in preventing osteoporosis and cardiovascular calcification. With the increasing market demand, the industrialization of vitamin K2 and the reduction of production costs have become the focus of attention. Vitamin K2 is currently produced mainly by chemical synthesis and biological fermentation. Due to the shortcomings of the chemical method, such as many by-products, low yield, and serious environmental pollution, and the presence of low-activity cis-isomers in the product, the product is not popular in the market. Highly active vitamin K2 can only be obtained through microbial fermentation, so fermentation production has a broader application prospect.

[0003] The prior art extracts vitamin K2 from fermentation broth mostly by organic solvent extraction, transferring vitamin K2 from the bacterial cell to an organic solvent, which is convenient for subsequent purification processes. Chinese patent CN109824496 discloses a method for extracting vitamin K2, wherein the fermentation broth is filtered and concentrated by a ceramic membrane, the bacterial cell is bombarded with a nitrogen ion beam to break the cell wall, and the cell wall is broken by adding 5-25 times the mass of ethanol and stirring extraction, and then the supernatant is collected by centrifugation to obtain a vitamin K2 extract. Chinese patent CN106631748 discloses a method for preparing vitamin K2, which uses centrifugal solid-liquid separation, wet bacterial cells are repeatedly frozen and thawed to break the cell wall, and n-hexane is added to stir and extract, and the vitamin K2 extract is further purified and separated by a resin column and a silica gel column, and then a pure K2 product is obtained by crystallization. Since the fermentation unit of vitamin K2 is very low (titer 100-200 mg / L), the bacterial content is not high after solid-liquid separation. In order to ensure the extraction yield, the traditional organic solvent stirring extraction process generally requires adding 4-6 times the solvent for 2-3 extractions, which results in an excessively large volume of the extract, a very low product content, and serious enrichment of other fat-soluble metabolites in the cell, making the subsequent purification process more difficult. Summary of the invention

[0004] The invention proposes a vitamin K2 extraction process, which solves the defects of the existing vitamin K2 extraction process that the amount of extraction solvent used is large and the extraction yield is low.

[0005] The technical solution of the present invention is achieved in this way:

[0006] A vitamin K2 extraction process comprises the following steps:

[0007] (1) adding a filter aid to the vitamin K2 fermentation broth, stirring evenly, and then performing solid-liquid separation to obtain a bacterial residue rich in vitamin K2;

[0008] (2) drying and crushing the fungus residue to obtain fungus powder;

[0009] (3) Loading the bacterial powder into the chromatography column and compacting the bacterial powder using clean gas;

[0010] (4) adding an organic solvent for extraction from the feed port of the chromatography column at a set flow rate, opening the discharge valve of the chromatography column, collecting the effluent from the chromatography column to obtain a high-concentration vitamin K2 extract, and after collecting 0.4-0.6 times the volume of the chromatography column, stopping the addition of the organic solvent, and pressing the liquid in the chromatography column with clean gas to dryness, and the extraction is completed;

[0011] (5) The high-concentration vitamin K2 extract is concentrated under vacuum to obtain a vitamin K2 ointment.

[0012] Furthermore, the filter aid in step (1) includes perlite or diatomaceous earth, and the mass of the added filter aid is 2% to 4% of the mass of the vitamin K2 fermentation broth.

[0013] Furthermore, the solid-liquid separation in step (1) is carried out by plate and frame filtration or centrifugal separation.

[0014] Furthermore, the mushroom residue in step (2) is dried by any one of air drying, vacuum drying and flash drying, and the drying temperature is 60-80°C.

[0015] Furthermore, the aspect ratio of the chromatography column in step (3) is 3:1 to 5:1.

[0016] Furthermore, the clean gas in step (3) and step (4) includes nitrogen.

[0017] Furthermore, the organic solvent in step (4) includes any one of ethanol, isopropanol, acetone, ethyl acetate, isopropyl acetate, butyl acetate, n-hexane, and n-heptane.

[0018] Furthermore, the organic solvent is ethyl acetate or acetone.

[0019] Furthermore, the addition flow rate of the organic solvent in step (4) is 0.25 to 1.0 BV / h.

[0020] Furthermore, the addition flow rate of the organic solvent is 0.5 BV / h.

[0021] Furthermore, the concentration in step (5) is vacuum concentration.

[0022] Beneficial effects of the present invention: The present invention innovatively uses a chromatography column to extract vitamin K2. Compared with the traditional stirring extraction process, it has the advantages of less extraction solvent usage, high extraction yield and high vitamin K2 ointment content, and is a production process with great industrial value. DETAILED DESCRIPTION

[0023] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0024] A vitamin K2 extraction process comprises the following steps:

[0025] (1) adding a filter aid to the vitamin K2 fermentation broth, stirring evenly, and then performing solid-liquid separation to obtain a bacterial residue rich in vitamin K2;

[0026] (2) drying and crushing the fungus residue to obtain fungus powder;

[0027] (3) Loading the bacterial powder into the chromatography column and compacting the bacterial powder using clean gas;

[0028] (4) adding an organic solvent for extraction from the feed port of the chromatography column at a set flow rate, opening the discharge valve of the chromatography column, collecting the effluent from the chromatography column to obtain a high-concentration vitamin K2 extract, and after collecting 0.4-0.6 times the volume of the chromatography column, stopping the addition of the organic solvent, and pressing the liquid in the chromatography column with clean gas to dryness, and the extraction is completed;

[0029] (5) The high-concentration vitamin K2 extract is concentrated under vacuum to obtain a vitamin K2 ointment.

[0030] Furthermore, the filter aid in step (1) includes perlite or diatomaceous earth, and the mass of the added filter aid is 2% to 4% of the mass of the vitamin K2 fermentation broth.

[0031] Furthermore, the solid-liquid separation in step (1) is carried out by plate and frame filtration or centrifugal separation.

[0032] Furthermore, the mushroom residue in step (2) is dried by any one of air drying, vacuum drying and flash drying, and the drying temperature is 60-80°C.

[0033] Furthermore, the aspect ratio of the chromatography column in step (3) is 3:1 to 5:1.

[0034] Furthermore, the clean gas in step (3) and step (4) includes nitrogen.

[0035] Furthermore, the organic solvent in step (4) includes any one of ethanol, isopropanol, acetone, ethyl acetate, isopropyl acetate, butyl acetate, n-hexane, and n-heptane.

[0036] Furthermore, the organic solvent is ethyl acetate or acetone.

[0037] Furthermore, the addition flow rate of the organic solvent in step (4) is 0.25 to 1.0 BV / h.

[0038] Furthermore, the addition flow rate of the organic solvent is 0.5 BV / h.

[0039] Furthermore, the concentration in step (5) is vacuum concentration.

[0040] Example 1

[0041] 30L of vitamin K2 fermentation broth, with a potency of 133.2mg / L, was added with 600g of perlite. After stirring evenly, plate and frame filtration was performed to obtain 3.26kg of filter cake.

[0042] The filter cake was dried by air blast for 26 h at a drying temperature of 60°C and crushed to obtain 1.89 kg of vitamin K2 bacterial powder.

[0043] After the bacterial powder was dry-transferred into a stainless steel chromatography column (3000 mL), it was compacted with compressed nitrogen.

[0044] Open the feed valve of the stainless steel chromatography column, add ethyl acetate at a flow rate of 1500mL / h, open the discharge valve, and start collecting the extract when liquid flows out; when about 1500mL of extract is collected, close the ethyl acetate feed valve, turn on the compressed nitrogen, press the ethyl acetate in the column dry, and combine to obtain 1860mL of high-concentration vitamin K2 extract with a potency of 2143.6mg / L.

[0045] The vitamin K2 high-concentration extract was concentrated to dryness in a vacuum at a temperature of 45° C. to obtain 36.7 g of vitamin K2 ointment with a mass content of 10.86% and a process yield of 99.8%.

[0046] Example 2

[0047] 500L of vitamin K2 fermentation broth, with a potency of 141.4mg / L, was added with 12kg of perlite, stirred evenly, and then plate and frame filtered to obtain 52.6kg of filter cake.

[0048] The filter cake was vacuum dried at 60°C for 18 h and crushed to obtain 33.2 kg of vitamin K2 bacterial powder.

[0049] The bacterial powder was dry-transferred into a stainless steel chromatography column (60 L), and then compacted with compressed nitrogen.

[0050] Open the feed valve of the stainless steel chromatography column, add acetone at a flow rate of 30 L / h, open the discharge valve, and start collecting the extract when liquid flows out; when about 30 L of extract is collected, close the ethyl acetate feed valve, turn on the compressed nitrogen, press the acetone in the column dry, and combine to obtain 38.6 L of high-concentration vitamin K2 extract with a potency of 1813.7 mg / L.

[0051] The vitamin K2 high-concentration extract was concentrated to dryness in a vacuum at a temperature of 50° C. to obtain 635.6 g of vitamin K2 ointment with a mass content of 11.02% and a process yield of 99.35%.

[0052] Comparative experiment

[0053] According to the method of Chinese patent CN109824496. 50L of vitamin K2 fermentation broth, with a potency of 127.2mg / L, add 1000g of perlite, stir evenly, filter with a plate and frame, and obtain 5.27kg of filter cake. Add 20L of ethanol to the filter cake, stir and extract for 4h, and filter to obtain 21.5L of primary extract with a potency of 221.1mg / L. Add 20L of ethanol again, stir and extract for 4h, and filter to obtain 20.6L of secondary extract with a potency of 62.3mg / L. Combine the two extracts of 42.1L, concentrate to dryness in a vacuum at 55℃, and obtain 292.6g of vitamin K2 ointment with a mass content of 2.17% and a process yield of 95.2%.

[0054] By comparison, it can be seen that the present invention uses a chromatography column to extract vitamin K2. Compared with the traditional stirring extraction process, it has the advantages of less extraction solvent usage, high extraction yield and high vitamin K2 ointment content, and is a production process with great industrial value.

[0055] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A process for extracting vitamin K2, characterized in that: The steps include: (1) adding a filter aid to the vitamin K2 fermentation broth, stirring evenly, and then performing solid-liquid separation to obtain a bacterial residue rich in vitamin K2; (2) drying and crushing the fungus residue to obtain fungus powder; (3) Loading the bacterial powder into the chromatography column and compacting the bacterial powder using clean gas; (4) adding an organic solvent for extraction from the feed port of the chromatography column at a set flow rate, opening the discharge valve of the chromatography column, collecting the effluent from the chromatography column to obtain a high-concentration vitamin K2 extract, and after collecting 0.4-0.6 times the volume of the chromatography column, stopping the addition of the organic solvent, and pressing the liquid in the chromatography column with clean gas to dryness, and the extraction is completed; (5) concentrating the high-concentration vitamin K2 extract to obtain a vitamin K2 ointment; The filter aid includes perlite or diatomaceous earth; the organic solvent includes any one of ethanol, isopropanol, acetone, ethyl acetate, isopropyl acetate, butyl acetate, n-hexane and n-heptane; and the addition flow rate of the organic solvent is 0.25-1.0 BV / h.

2. The process for extracting vitamin K2 according to claim 1, characterized in that: The mass of the filter aid added in step (1) is 2% to 4% of the mass of the vitamin K2 fermentation broth.

3. The process for extracting vitamin K2 according to claim 1, characterized in that: The solid-liquid separation in step (1) is carried out by plate and frame filtration or centrifugal separation.

4. The process for extracting vitamin K2 according to claim 1, characterized in that: The mushroom residue in step (2) is dried by any one of air drying, vacuum drying and flash drying, and the drying temperature is 60-80°C.

5. The process for extracting vitamin K2 according to claim 1, characterized in that: The height-to-diameter ratio of the chromatography column in step (3) is 3:1 to 5:

1.

6. The process for extracting vitamin K2 according to claim 1, characterized in that: The clean gas in step (3) and step (4) includes nitrogen.

7. The process for extracting vitamin K2 according to claim 1, characterized in that: The addition flow rate of the organic solvent is 0.5 BV / h.

8. The process for extracting vitamin K2 according to claim 1, characterized in that: The concentration in step (5) is vacuum concentration.