Bacillus subtilis natto and method for purifying vitamin menadione-7 by using bacterial strain

A technology for vitamin A and natto subtilis, which is applied in the field of purifying vitamin menadione-7, and achieves the effects of simplifying purification steps, improving purity and improving efficiency

Active Publication Date: 2015-01-07
SUNGEN BIOSCIENCE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0019] However, at present, there is no simple and efficient method for purifying vitamin K2, especially the method for purifying MK-7 of vitamin K2.

Method used

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  • Bacillus subtilis natto and method for purifying vitamin menadione-7 by using bacterial strain
  • Bacillus subtilis natto and method for purifying vitamin menadione-7 by using bacterial strain
  • Bacillus subtilis natto and method for purifying vitamin menadione-7 by using bacterial strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Preparation of Bacillus natto fermentation broth:

[0063] Natto (purchased from Japan Biotechnology Co., Ltd.) was dissolved and diluted with sterile water and spread on an agarose-fibrin plate (see: Wei Hua, Zhao Xiangying, Liu Jianjun, Determination of Nattokinase Activity [J]. Shandong Journal of the Institute of Light Industry, 2007, 21(1) 60-63.), after culturing at 37°C for 18 hours, a transparent circle was produced, and colonies with larger transparent circles were selected for multi-generation mutagenesis by ultraviolet light (15s) to obtain Bacillus subtilis natto (Bacillus subtilis natto) ST188 (strains are preserved in the General Microbiology Center of China Microbiological Culture Collection Management Committee, address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, preservation date: October 31, 2013, and preservation number is : CGMCC No.8400).

[0064] Inoculate 500 mL of the obtained Bacillus subtilis ST188 in natto into 10000 mL of ...

Embodiment 2

[0074]The method of Example 1 was used to purify vitamin MK-7 in Bacillus natto fermentation broth, the difference was that in step (4), the volume ratio of wet packing to dry packing was 2:1.

[0075] The H-spectrum, C-spectrum, DEPT135-spectrum, COZY-spectrum, HSQC-spectrum and HMBC-spectrum of the product obtained in Example 1 are consistent, so it can be confirmed that the product is vitamin menadione-7.

[0076] At the same time, the purity of the vitamin MK-7 in the product is determined to be 99% through chromatographic analysis, and the yield of the obtained vitamin MK-7 is 93%.

Embodiment 3

[0078] The method in Example 1 was used to purify vitamin MK-7 in Bacillus natto fermentation broth, the difference was that in step (4), the volume ratio of wet packing to dry packing was 1:2.

[0079] The H-spectrum, C-spectrum, DEPT135-spectrum, COZY-spectrum, HSQC-spectrum and HMBC-spectrum of the product obtained in Example 1 are consistent, so it can be confirmed that the product is vitamin menadione-7.

[0080] At the same time, the purity of the vitamin MK-7 in the product is determined to be 93% through chromatographic analysis, and the yield of the obtained vitamin MK-7 is 92%.

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Abstract

The invention provides bacillus subtilis natto ST188 with CGMCC No. 8400 and a high-yield method for purifying vitamin menadione-7 by using the bacterial strain. The high-yield method comprises the following steps: (1) carrying out spray drying on natto fermentation liquor, and extracting or leaching bacillus natto fermentation liquor spray drying powder by using a solvent so as to obtain a leaching solution; (2) concentrating the leaching solution obtained in the step (1) so as to obtain extract; (3) carrying out column chromatography on the extract obtained in the step (2), carrying out gradient or isocratic elution, and concentrating collected liquid, thus obtaining a menadione-7 crude product; and (4) crystallizing and purifying the menadione-7 crude product obtained in the step (3), thus obtaining the pure vitamin menadione-7 product.

Description

[0001] This application claims the priority of the Chinese patent application number 201410129217.2 submitted on March 31, 2014, and the invention title is "A Bacillus subtilis natto and a method for purifying vitamin menadione-7 from the strain" . technical field [0002] The invention relates to a Bacillus subtilis natto ST188, CGMCC No.8400, and a method for purifying vitamin menadione-7 from the strain. Background technique [0003] Because vitamin K has the effect of promoting blood clotting, it is also called blood clotting vitamin. For nearly half a century, people's understanding of vitamin K was limited to its effect on the blood coagulation system, until Bouckaert discovered in 1960 that vitamin K could promote bone fracture healing in rabbits. In 1975, after Pettifor first put forward the hypothesis that vitamin K participates in human bone metabolism, people have a deep understanding of the role of vitamin K in bone metabolism. [0004] The vitamin K family inc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07C50/14C07C46/10C12N1/20C12R1/125
CPCC07C46/10C12N1/205C12R2001/125C07C50/14
Inventor 陈杰鹏段丽丽陈煜藩陈鸿锐邱雪莲蔡桂珠洪琳胡留松黄晓莹张岳霖许志锴
Owner SUNGEN BIOSCIENCE CO LTD
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