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Method for separating and purifying coenzyme Q10 from microorganism

A separation and purification, microorganism technology, applied in microorganism-based methods, biochemical equipment and methods, quinone separation/purification, etc., can solve the problems of difficult solvent recovery, low efficiency of bacterial cell fragmentation, and difficult separation, etc. Extraction rate and yield, low production cost, low environmental pollution effect

Active Publication Date: 2014-05-28
湖南莱崔尔生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] According to the above process to produce coenzyme Q10, the efficiency of cell fragmentation is low, and the types and quantities of organic solvents are more, and the operation is more complicated.
In the chromatographic process, n-hexane and another hydrophobic organic solvent are used as eluents, which are not easy to separate and cause great difficulties for subsequent solvent recovery.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] This embodiment includes the following steps:

[0023] (1) Broken: Rhodococcus bacteria (purchased from CGMCC, preservation number CGMCCNO. 1.2569) were fermented to obtain 5 L of fermentation broth, and the filter cake was collected by filtration to obtain 782 g of wet bacteria. Take 500 g of the collected wet bacteria After resuspending with 1.5 mol / L hydrochloric acid solution equivalent to 5 times the volume of wet bacteria, the method of ultrasonic crushing was used to assist the cell wall breaking. The conditions of ultrasonic crushing were power 500 W, frequency 0.6, and ultrasonic crushing twice. , 10 minutes each time, to obtain the bacterial suspension;

[0024] (2) Extraction: Add NaOH solution to adjust the bacterial suspension obtained in step (1) to pH 7.0, and extract twice with a mixed solution of ethyl acetate:petroleum ether=7:93, and add the volume of organic solvent for each extraction: Bacterial suspension volume = 2:1, stirring and extracting for ...

Embodiment 2

[0034] This embodiment includes the following steps:

[0035] The crude extract described in Example 1, after dehydration treatment, can be used for silica gel column chromatography; take 50 g of dried silica gel and pack it into a column, and equilibrate; take 1000 ml of the crude extract, inject it into a well-balanced silica gel column, and put it on The column flow rate is 1 BV / h. After loading the column, wash with 1 times the column volume of petroleum ether, and then elute with petroleum ether containing 7% ethyl acetate at a flow rate of 1 BV / h. Collect the eluted Liquid 635 ml, adopt the HPLC detection method of coenzyme Q10 described in Example 1 to detect. The content of coenzyme Q10 in the eluate reached 92.8%, and the extraction rate was 94.4%.

Embodiment 3

[0037] This embodiment includes the following steps:

[0038] The eluate obtained in Example 2 was concentrated under reduced pressure to a ratio of coenzyme Q10: organic solvent of 1:10. After the concentrated solution is cooled to 30°C, add 1 wt% of the coenzyme Q10 content in the concentrated solution to the seed crystal, stir at 50rpm, keep the temperature at a constant temperature for 30 minutes, then cool down to 20°C at 6°C / h, maintain for half an hour, and then filter to obtain After the crystals are washed and dried, the refined coenzyme Q10 product can be obtained. The HPLC detection method for coenzyme Q10 described in Example 1 is used for detection, and the purity of the obtained coenzyme Q10 product can reach 98.4%, and the total yield from the bacteria to the product is 82.7%.

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Abstract

A method for separating and purifying coenzyme Q10 from a microorganism comprises the following steps of: (1) crushing, (2) extracting, (3) chromatography and (4) crystallization. With the adoption of the method for preparing the coenzyme Q10, the extraction rate and yield of the coenzyme Q10 can be guaranteed, and the quality conformance of a coenzyme Q10 product can be further ensured. The method is simple in technology, low in production cost and small in environmental pollution and is appropriate for industrial production.

Description

technical field [0001] The invention relates to a method for isolating and purifying coenzyme Q10, in particular to a method for isolating and purifying coenzyme Q10 from microorganisms. Background technique [0002] Coenzyme Q10 (Coenzyme Q10, CoQ10), molecular formula C 59 h 90 o 4 , relative molecular weight 863.36, easily soluble in most organic reagents, slightly soluble in ethanol, insoluble in methanol and water, easy to photodecompose, relatively stable to humidity and temperature. [0003] Coenzyme Q10 is an electron transporter in the respiratory chain, has anti-oxidation function, and is widely used in the comprehensive treatment of cardiovascular diseases, hepatitis, cancer, AIDS, etc. and to improve human immunity. [0004] At present, there are three methods for producing coenzyme Q10: animal and plant tissue extraction, microbial fermentation and chemical synthesis. Among them, the production of coenzyme Q10 by microbial method has the characteristics of e...

Claims

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Application Information

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IPC IPC(8): C07C50/28C07C46/10C12P7/66C12R1/01
CPCC07C46/10C07C50/28
Inventor 陶帅刘志强
Owner 湖南莱崔尔生物科技有限公司
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