Method for enriching vitamin K2 in natto bacillus fermentation liquid

A technology of Bacillus natto and fermented liquid, which is applied in the field of bioengineering and bioseparation engineering, can solve the problems of high difficulty in purifying vitamin K, complicated overall process, and high cost, and achieves cheap and easy-to-obtain raw materials, fast separation rate, The effect of reducing dosage

Pending Publication Date: 2019-05-03
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

[0004] Preparation of Vitamin K by Fermentation of Bacillus Natto 2 Among them, Bacillus natto has more metabolites, and at the same time, due to its cell autolysis, it will produce a variety of impurities, such as miscellaneous proteins, nucleic acids, oils, etc., resulting in the acquisition of highly purified vitamin K 2 more difficult
[0005] Currently, organic solvents are widely used to extract vitamin K from Bacillus natto fermentation broth 2 , this method requires pretreatment operations such as bacterial crushing, dehydration and drying, and subsequent purification methods also require macroporous resin and reversed-phase silica gel, etc. The overall process is complex, expensive, and the extraction rate is low

Method used

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  • Method for enriching vitamin K2 in natto bacillus fermentation liquid

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Embodiment 1

[0033] Embodiment 1: Vitamin K in a kind of enrichment Bacillus natto fermented liquid 2 Methods

[0034] (1) Obtaining the supernatant of the fermentation broth

[0035] The fermented liquid processed in this embodiment is 69.6g / L by glycerol, glucose is 34.5g / L, K 2 HPO 4 4.0g / L peptone, 20g / L peptone, 25g / L yeast extract, 250rpm, cultured at 37°C for five days to obtain Bacillus natto fermentation broth.

[0036]The above fermented liquid of Bacillus natto was centrifuged and filtered to remove bacteria and sediment to obtain a supernatant.

[0037] (2) Flocculant purification of fermentation broth

[0038] First add acetic acid to water to form a 0.18%wt acetic acid solution, then add chitosan to the 0.18%wt acetic acid solution to form a 0.4%wt chitosan acetic acid solution.

[0039] Take 100m of the above-mentioned supernatant from the removal of bacteria, add 9g of the above-mentioned chitosan acetic acid solution to it, stir for 30min, finally add filter aid, disc...

Embodiment 2

[0052] Embodiment 2 A kind of vitamin K enriched in Bacillus natto fermented liquid 2 Methods

[0053] A kind of vitamin K enriched in Bacillus natto fermented liquid 2 method, including the following steps:

[0054] Step 1: Centrifuge the fermented liquid of Bacillus natto, and collect the supernatant;

[0055] Step 2: Add flocculant and filter aid to the supernatant obtained in step 1, then stir, then centrifuge, and collect the supernatant;

[0056] Step 3: Add reduced Fe to the supernatant from Step 2 3 o 4 @SiO 2 -G nano-microspheres, adjust the pH value to 6.0, then carry out stirring treatment at 20°C for 10minmin, and then collect the magnetic material after adsorption; the reduced Fe 3 o 4 @SiO 2 -The preparation method of G nano microspheres comprises the following steps:

[0057] S1, the Fe 3 o 4 The magnetic microspheres were added to the mixed solution composed of deionized water, isopropanol ammonia water and tetraethyl orthosilicate, stirred evenly an...

Embodiment 3

[0067] Embodiment 3: Vitamin K in a kind of enrichment Bacillus natto fermented liquid 2 Methods

[0068] A kind of vitamin K enriched in Bacillus natto fermented liquid 2 method, including the following steps:

[0069] Step 1: Centrifuge the fermented liquid of Bacillus natto, and collect the supernatant;

[0070] Step 2: Add flocculant and filter aid to the supernatant obtained in step 1, then stir, then centrifuge, and collect the supernatant;

[0071] Step 3: Add reduced Fe to the supernatant from Step 2 3 o 4 @SiO 2 -G nano-microspheres, adjust the pH value to 8.0, then carry out stirring treatment at 35°C for 30min, and then collect the magnetic material after adsorption; the reduced Fe 3 o 4 @SiO 2 -The preparation method of G nano microspheres comprises the following steps:

[0072] S1, the Fe 3 o 4 The magnetic microspheres were added to a mixed solution composed of deionized water, isopropanol ammonia and tetraethyl orthosilicate, stirred evenly and reacte...

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Abstract

The invention relates to a method for enriching vitamin K2 in natto bacillus fermentation liquid. The method comprises the following steps of step 1, performing centrifugation on the natto bacillus fermentation liquid, and collecting supernatant; step 2, adding a flocculant and a filter aid to the supernatant obtained in the step 1, then performing stirring, performing centrifugation, and collecting supernatant; step 3, adding Fe3O4@SiO2-G nanometer microspheres which are reduced to the supernatant obtained in the step 2, regulating the pH to 6.0-8.0, then performing stirring treatment under the condition of 20-35 DEG C for 10-30min, and then collecting magnetic materials after adsorption; and step 4, adding hexane to the magnetic materials after adsorption, performing ultrasonic parsing for 5-20min, then externally applying a magnet, collecting magnetic nanometer materials, pouring out organic phase, and performing filtering, wherein filtrate is a hexane solution rich in vitamin K2. According to the method for enriching vitamin K2 in natto bacillus fermentation liquid, the VK2 in the natto bacillus fermentation liquid can be quickly and efficiently enriched. The method has the advantages that the specific steps are simple, the materials are low in price, and can be reused, and the enrichment effect is efficient.

Description

technical field [0001] The invention relates to a method for enriching vitamin K in Bacillus natto fermentation liquid 2 The method belongs to the fields of bioengineering and bioseparation engineering. Background technique [0002] Vitamin K 2 It is a general term for a series of terpene side chain compounds containing 2-methyl-1, 4-naphthoquinone core and C3 with different numbers of isoprene structural units. According to the carbon element on the terpene side chain The number can be divided into K2 (10), K2 (20), K2 (35), K2 (40) and so on. Vitamin K 2 It is a fat-soluble vitamin, a derivative of a naphthoquinone group with biological activity of phylloquinone, and is one of the indispensable important vitamins in the human body. With research on its function, it has been proven that vitamin K 2 It has the effect of preventing osteoporosis, cardiovascular, Parkinson, liver cancer and other diseases. Therefore, functional foods, medical products, cosmetics and other...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/66C12R1/07
Inventor 郑之明方志伟王鹏王晗王丽刘会赵根海吴荷芳孙小雯倪文枫杨强唐恒芳
Owner HEFEI INSTITUTES OF PHYSICAL SCIENCE - CHINESE ACAD OF SCI
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