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
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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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