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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

Examples

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Embodiment 1

[0041] Vitamin K isolated and purified from Bacillus natto 2 method, such as figure 1 shown, including the following steps:

[0042] (1) Vitamin K 2 Extraction

[0043] Taking Bacillus natto fermentation broth as vitamin K 2 Extract the original solution, centrifuge to obtain the bacteria, after drying, use ethanol to extract to obtain vitamin K 2 Extraction solution; the extraction method is static extraction, the extraction time is 20min, and the extraction frequency is 1 time.

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

[0045] Vitamin K 2 The extract is passed through a membrane separation device (membrane pore size ≤ 0.45 μm) to remove insoluble impurities, and the obtained vitamin K 2 The filtrate was evaporated to dryness under reduced pressure, and then redissolved with methanol to obtain vitamin K 2 Stock solution on the column;

[0046] (3) Low-purity vitamin K 2 Acquisition of Concentrate

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

Embodiment 2

[0057] Vitamin K isolated and purified from Bacillus natto 2 method, such as figure 1 shown, including the following steps:

[0058] (1) Vitamin K 2 Extraction

[0059] Taking Bacillus natto fermentation broth as vitamin K 2 Extract the original 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 60min, and the extraction frequency is 3 times.

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

[0061] Vitamin K 2 The extract is passed through a membrane separation device (membrane pore size ≤ 0.45 μm) to remove insoluble impurities, and the obtained vitamin K 2 The filtrate was evaporated to dryness under reduced pressure, then redissolved with acetone to obtain vitamin K 2 Stock solution on the column;

[0062] (3) Low-purity vitamin K 2 Acquisition of Concentrate

[0063] Vitamin K 2 The stock solution on the...

Embodiment 3

[0073] Vitamin K isolated and purified from Bacillus natto 2 method, such as figure 1 shown, including the following steps:

[0074] (1) Vitamin K 2 Extraction

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

[0076] ① Weigh several portions of 1.0g of dry bacteria into a 50ml centrifuge tube, add 5ml of different organic solvents to stand for extraction for 50min, extract twice, 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] ② Dilute the obtained extract to 5ml to obtain the test solution;

[0078] ③Use the 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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