Method for separating and purifying vitamin K2 in bacillus subtilis natto

A technology for separation and purification of Bacillus natto is applied in the field of bioengineering to achieve the effects of high biological activity, simple process operation and convenient industrial application.

Active Publication Date: 2017-05-10
HEFEI INSTITUTES OF PHYSICAL SCIENCE - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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  • Method for separating and purifying vitamin K2 in bacillus subtilis natto
  • Method for separating and purifying vitamin K2 in bacillus subtilis natto
  • Method for separating and purifying vitamin K2 in bacillus subtilis natto

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[0040] Example 1

[0041] A kind of separation and purification of vitamin K from Bacillus natto 2 Method such as figure 1 As shown, including the following steps:

[0042] (1) Vitamin K 2 Extraction

[0043] Using Bacillus natto fermentation broth as vitamin K 2 Extract the stock solution, centrifuge to obtain the bacteria, after drying, extract with ethanol to obtain vitamin K 2 Extraction solution: The extraction method is static extraction, the extraction time is 20 minutes, and the number of extractions is once.

[0044] (2) Vitamin K 2 Obtaining the upper column stock solution

[0045] Vitamin K 2 The extract is passed through a membrane separation device (membrane pore size≤0.45μm) to remove insoluble impurities, and the obtained vitamin K 2 The filtrate is evaporated to dryness under reduced pressure, and then re-dissolved in methanol to obtain vitamin K 2 Upper column stock solution;

[0046] (3) Low purity vitamin K 2 Obtaining concentrate

[0047] Vitamin K 2 The stock solution...

Example Embodiment

[0056] Example 2

[0057] A kind of separation and purification of vitamin K from Bacillus natto 2 Method such as figure 1 As shown, including the following steps:

[0058] (1) Vitamin K 2 Extraction

[0059] Using Bacillus natto fermentation broth as vitamin K 2 Extract the stock solution, centrifuge to obtain the bacteria, after drying, extract with toluene to obtain vitamin K 2 Extraction solution: The extraction method is static extraction, the extraction time is 60 minutes, and the extraction times are 3 times.

[0060] (2) Vitamin K 2 Obtaining the upper column stock solution

[0061] Vitamin K 2 The extract is passed through a membrane separation device (membrane pore size≤0.45μm) to remove insoluble impurities, and the obtained vitamin K 2 The filtrate is evaporated to dryness under reduced pressure, and then re-dissolved in acetone to obtain vitamin K 2 Upper column stock solution;

[0062] (3) Low purity vitamin K 2 Obtaining concentrate

[0063] Vitamin K 2 The stock solution o...

Example Embodiment

[0072] Example 3

[0073] A kind of separation and purification of vitamin K from Bacillus natto 2 Method such as figure 1 As shown, including the following steps:

[0074] (1) Vitamin K 2 Extraction

[0075] Take Bacillus natto fermentation broth (composition: soybean meal 65g / L, corn meal 60g / L, peptone 50g / L, fermentation period: 7-9 days) as vitamin K 2 Extract the stock solution, centrifuge to obtain the bacteria, and after drying, extract according to the following steps:

[0076] ①Weigh several aliquots of 1.0g dry cells in a 50ml centrifuge tube, add 5ml different organic solvents to stand for 50min, extract 2 times, and make 3 parallel samples; the different organic solvents are methanol, ethanol, 1,3 propylene glycol, n-butanol, acetic acid, dichloromethane, isoamyl alcohol, n-hexane, ethyl acetate, chloroform, carbon tetrachloride, acetone, ether, toluene;

[0077] ②Fix the volume of the obtained extract to 5ml to obtain the test solution;

[0078] ③Use HPLC method (mobile ph...

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Abstract

The invention discloses a method for separating and purifying vitamin K2 in bacillus subtilis natto. The method comprises the following steps: acquiring vitamin K2 extract, acquiring a vitamin K2 column stock solution, acquiring a low-purity vitamin K2 concentrated solution, acquiring a high-purity vitamin K2 concentrated solution, acquiring high-purity vitamin K2, and acquiring vitamin K2 crystals. The method has the advantages that (1) an obtained vitamin K2 crystal product is high in purity, bioactivity and recovery rate; (2) a separation and purification process is easy to operate, mild in condition and high in treatment capability; (3) each organic solvent can be recycled for industrial application.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a method for separating and purifying vitamin K from Bacillus natto 2 Methods. Background technique [0002] Vitamin K 2 (Menaquinone, MK, VK 2 ) is a general term for a class of menaquinone compounds, the chemical structural formula is 2-methyl-3-enyl-1,4-naphthoquinone, according to the number of isoprene units on the C-3 side chain, MK can be Divided into 14 kinds, represented by MK-n (n=1-14). Studies have shown that vitamin K 2 It can not only promote blood coagulation, increase osteocalcin synthesis, but also reduce the risk of liver cancer and prostate cancer. Recent studies have shown that vitamin K 2 It can also be used as an adjuvant treatment for Parkinson's syndrome and amyotrophic lateral sclerosis. The safety of vitamin K2 has been recognized by the China Food and Drug Administration, the American Institute of Drug Research, the US FDA, the Council of ...

Claims

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Application Information

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IPC IPC(8): C07C46/10C07C50/14
CPCC07C46/10C07C50/14
Inventor 郑之明尉鸿飞王鹏王丽王晗倪文枫吴荷芳赵根海刘会
Owner HEFEI INSTITUTES OF PHYSICAL SCIENCE - CHINESE ACAD OF SCI
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