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Method for separating and purifying vitamin K2 in fermentation liquor by using palm oil extract

A technology for separation and purification of vitamins, applied in quinone separation/purification, organic chemical methods, organic chemistry, etc., can solve the problems that the product cannot be directly consumed by the human body, serious pollution, complicated operation, etc., and achieve good mutual solubility, low freezing point, Good oxidation stability

Active Publication Date: 2021-06-22
HEFEI INSTITUTES OF PHYSICAL SCIENCE - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the separation and purification solvents used in the past are methanol, tetrahydrofuran, dichloromethane and other highly toxic organic reagents as solvents. Multi-step operations are required to obtain vitamin K2 with high purity. During this process, the pollution is serious and a large amount of waste liquid is generated. The operation is complicated, the cost is high, and the obtained product cannot be directly used for human consumption

Method used

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  • Method for separating and purifying vitamin K2 in fermentation liquor by using palm oil extract
  • Method for separating and purifying vitamin K2 in fermentation liquor by using palm oil extract
  • Method for separating and purifying vitamin K2 in fermentation liquor by using palm oil extract

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Effect test

Embodiment 1

[0032] Embodiment 1: A kind of method utilizing palm oil extract to isolate and purify vitamin K2 in fermented liquid

[0033] Using palm oil extract (medium-chain triglyceride) as a solvent to extract vitamin K2 from dry bacteria in one step, and to separate and purify high-purity vitamin K2 in two steps, mainly including the following steps:

[0034] (1) Strain seed culture. Bacillus natto or Bacillus subtilis needs to be cultured with seed liquid to obtain a certain amount of pure strain liquid before fermentation.

[0035] (2) Preparation of fermentation medium. Continuously optimize the composition of the fermentation medium through the feedback results of the fermentation experiment, and select the best fermentation medium formula for fermentation.

[0036] (3) Regulation of the fermentation process. Inoculate the seed solution in step (1) into the fermentation medium configured in step (2). The fermentation temperature is 35-40°C, and the rotation speed is kept at 2...

Embodiment 2

[0071] Embodiment 2: A kind of method utilizing palm oil extract to isolate and purify vitamin K2 in fermented liquid

[0072] Firstly, Bacillus natto is fermented, and the fermented wet cells are freeze-dried to obtain dry cells of Bacillus natto, and palm oil extract (medium chain triglyceride) is used as a solvent to extract the dry cells of Bacillus natto vitamin K2, and then use green non-toxic ethanol and butyl acetate as solvents for separation and purification, and obtain high-purity vitamin K2 in a green and efficient two-step method. details as follows:

[0073] The bacillus natto was fermented and cultivated in a 30L stirred reactor, and the filling volume was 80%. Inoculate the seed culture solution with 10% inoculum amount, ferment at 37°C and 250rpm, and monitor the bacterial concentration (OD method) during the cultivation until 156 hours later, the cultivation ends. Centrifuge the Bacillus natto or Bacillus subtilis fermentation liquid obtained by the microbi...

Embodiment 3

[0078] Embodiment 3: A kind of method utilizing palm oil extract to isolate and purify vitamin K2 in fermented liquid

[0079] A method utilizing palm oil extract to separate and purify vitamin K2 in fermented liquid, comprising the following steps:

[0080] Step 1: Separating the fermented liquid obtained by the microbial fermentation production method to obtain wet cells, and then freeze-drying the wet cells to obtain dry cells rich in vitamin K2;

[0081] Step 2: Mix the dried bacteria with the palm oil extract for 2 hours, then centrifuge the mixture to take the supernatant, and the obtained supernatant is the palm oil extract rich in vitamin K2;

[0082] Step 3: Pack the pretreated macroporous resin into a column, place the vitamin K2-rich palm oil extract in a separation column filled with macroporous resin, use the first eluent to remove miscellaneous oils, and then use the second A resolving agent elutes vitamin K2 from the macroporous resin to obtain an eluent;

[008...

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Abstract

The invention relates to a method for separating and purifying vitamin K2 in fermentation liquor by using palm oil extract, which comprises the following steps: separating the fermentation liquor to obtain wet thalli, conducting freeze-drying to obtain dry thalli, mixing the dry thalli with palm oil extract for 2-4h, and conducting centrifuging to take supernate; packing pretreated macroporous resin into a column, putting the supernate into the separation column filled with the macroporous resin, removing miscellaneous oil substances by using a first eluent, and eluting the vitamin K2 from the macroporous resin by using a first desorption agent to obtain an eluate; concentrating the obtained eluate to obtain a concentrated solution; putting the concentrated solution into a separation column containing reverse phase silica gel, removing impurities by using a second eluent, conducting eluting by using a second desorption agent to obtain a high-purity vitamin K2 product, and conducting freezing and crystallizing to obtain vitamin K2 crystals. The palm oil extract used in the invention exists in the nature, is relatively simple to obtain and does not generate organic waste liquid, so that the pollution to the environment is greatly reduced.

Description

technical field [0001] The invention belongs to the field of biochemical engineering, and relates to a method for separating and purifying vitamin K2 in fermented liquid by using palm oil extract, in particular to using palm oil extract (such as medium-chain triglyceride) and butyl acetate as solvent green A method for efficiently extracting vitamin K2 in Bacillus natto and Bacillus subtilis. Background technique [0002] Vitamin K2 is a series of derivatives of 2-methyl-3-alkyl-1,4-naphthoquinone. According to the number of isoprene at the C-3 position, it can be divided into 14 types. MK-n Indicates (n refers to the number of isoprene units on the side chain). With the in-depth research on the function of vitamin K2, it is found that it can not only promote blood coagulation, prevent and treat osteoporosis, but also have a good preventive effect on neurodegenerative diseases and various tumors. In Japan, Europe and the United States, it has already been sold as functiona...

Claims

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Application Information

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IPC IPC(8): C07C46/10C07C50/14
CPCC07C46/10C07B2200/13C07C2602/10C07C50/10
Inventor 郑之明马国良王鹏王丽王晗赵根海丁秀敏杨强
Owner HEFEI INSTITUTES OF PHYSICAL SCIENCE - CHINESE ACAD OF SCI
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