Equipment and method for extracting vitamin K2 from microbial fermentation liquor

A technology for microbial fermentation broth and vitamins, applied in organic chemistry, quinone separation/purification, etc., can solve problems such as cumbersome process, low purity and/or yield of vitamin K2, achieve high concentration, large sample volume, and reduce production cost effect

Active Publication Date: 2021-02-05
INNER MONGOLIA KINGDOMWAY PHARMA LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] The purpose of the present invention is to overcome the defects that the purity and / or yield of vitamin K2 obtained by the method for extracting vitamin K2 (MK-7) from microbial fermentation broth is relatively low and the process is cumbersome, and to provide a method that can simultaneously The equipment and method for extracting vitamin K2 from microbial fermentation broth to improve the purity and yield of vitamin K2 (MK-7). The entire extraction process can be fully automated, with good process stability, fast separation speed, and short time-consuming. Continuous operation can well make up for the deficiencies in the existing technology that the extraction and purification process is long, high cost, low yield, manual intervention is required, and it is not suitable for industrial production

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  • Equipment and method for extracting vitamin K2 from microbial fermentation liquor
  • Equipment and method for extracting vitamin K2 from microbial fermentation liquor

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Embodiment 1

[0049] Embodiment 1 (reversed-phase chromatographic column)

[0050] In this embodiment, vitamin K2 (MK-7) is separated, extracted and purified from microbial fermentation broth by means of continuous dynamic countercurrent extraction combined with medium-pressure preparative chromatographic separation and purification. The specific steps are as follows:

[0051] (1) 50 L of the vitamin K2 stock solution produced by Bacillus subtilis fermentation was subjected to wall-breaking treatment with concentrated sulfuric acid, and then solid-liquid separation was performed by a tubular centrifuge to obtain 3.0 kg of wet sludge. The wet sludge and acetone-ethanol mixed solvent (v:v=1:2) are subjected to dynamic countercurrent extraction in a continuous dynamic countercurrent extractor (length-to-diameter ratio is 30:1) according to the mass volume ratio of 1kg:10L, and the extraction is controlled. The temperature is 40° C., the extraction residence time is 1 h, and an extract solution...

Embodiment 2

[0054] Embodiment 2 (reversed-phase chromatographic column)

[0055] In this embodiment, vitamin K2 (MK-7) is extracted and purified from microbial fermentation broth by means of continuous dynamic countercurrent extraction combined with medium-pressure preparative chromatographic separation and purification. The specific steps are as follows:

[0056] (1) 50 L of the vitamin K2 stock solution produced by the fermentation of Bacillus subtilis was broken with concentrated hydrochloric acid, and then solid-liquid separation was performed with a tubular centrifuge to obtain 3.2 kg of wet sludge. The wet sludge and methanol-dichloromethane mixed solvent (v:v=3:1) were subjected to dynamic countercurrent extraction in a continuous dynamic countercurrent extractor (length-to-diameter ratio of 100:1) at a mass volume ratio of 1kg:8L, The extraction temperature was controlled at 30° C., the extraction residence time was 1 h, and an extract solution with a vitamin K2 (MK-7) concentrati...

Embodiment 3

[0059] Embodiment 3 (reversed-phase chromatographic column)

[0060] In this embodiment, vitamin K2 (MK-7) is separated, extracted and purified from microbial fermentation broth by means of continuous dynamic countercurrent extraction combined with medium-pressure preparative chromatographic separation and purification. The specific steps are as follows:

[0061] (1) 3m3 of the vitamin K2 stock solution produced by the fermentation of Bacillus subtilis was subjected to wall-breaking treatment with acetic acid, and then solid-liquid separation was carried out by a butterfly centrifuge to obtain 180 kg of wet sludge. The wet sludge and methanol-ethyl acetate mixed solvent (v:v=3:1) were subjected to dynamic countercurrent extraction in a continuous dynamic countercurrent extractor (length-to-diameter ratio of 60:1) at a mass volume ratio of 1kg:10L, The extraction temperature was controlled at 35° C., the extraction residence time was 1.5 h, and an extract with a vitamin K2 (MK-...

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Abstract

The invention belongs to the technical field of microbial extraction, and relates to equipment and a method for extracting vitamin K2 from microbial fermentation liquor. The equipment comprises a continuous dynamic countercurrent extractor and a medium-pressure preparative chromatography, the medium-pressure preparative chromatography comprises at least one chromatographic group, each chromatographic group comprises four chromatographic columns connected in parallel, each chromatographic column is independently provided with a feed port, an eluent inlet A, an eluent inlet B and an eluent inletC, the feed port is connected with an extracting solution outlet of the continuous dynamic countercurrent extractor, the eluent inlet A, the eluent inlet B and the eluent inlet C are respectively connected with an eluent source A, an eluent source B and an eluent source C, the opening and closing of the four material inlets are respectively and independently controlled by valves, and the valves of different chromatographic columns in the same chromatographic group are synchronously switched. By adopting the equipment provided by the invention, the purity and yield of the vitamin K2 can be improved, and the whole extraction process can completely realize continuous automatic operation.

Description

technical field [0001] The invention belongs to the technical field of microorganism extraction, and in particular relates to a device and a method for extracting vitamin K2 from microorganism fermentation liquid. Background technique [0002] Vitamin K2, also known as menaquinone (Menaquinone), is usually represented by MK. It consists of a group of compounds with a total of 14 forms, the difference lies in the length of the side chain, among which MK-7 (menadione-7) has the most significant biological activity. The chemical formula of MK-7 is shown in formula (1). It contains 7 isoprene units on the side chain of vitamin K2. It is an important class of fat-soluble coagulation vitamins and is a rare oil resource. The content in it is very small, and it is known as "platinum vitamin". It has many important physiological functions such as prevention and treatment of osteoporosis, arterial calcification, cardiovascular disease, tumor and Parkinson's disease. [0003] [0...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07C50/10C07C46/10
CPCC07C46/10C07C2602/10C07C50/14
Inventor 王丽廖炜程王海彬刘纯旭胡泽君
Owner INNER MONGOLIA KINGDOMWAY PHARMA LTD
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