Method for fermentation preparation of cozymase Q10
A coenzyme and fermentation liquid technology, applied in the direction of microorganism-based methods, methods using bacteria, biochemical equipment and methods, etc., can solve the problems of high production costs, and achieve the effects of low production costs, simple processes, and high bacterial concentration
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Embodiment 1
[0039] Inoculate Alcaligenes eutropha H16.65-7 into a culture medium with the following composition (in g / L) at 10%: glucose 20, ammonium sulfate 1.8, dipotassium hydrogen phosphate 1.5, disodium hydrogen phosphate 9, Magnesium sulfate 2, yeast extract 3, peptone 2, PH6.8-7.0.
[0040] Then, carry out high-density fermentation under the following control conditions: sugar concentration 2-2.5%, temperature 30°C, air volume 0.3-1VVM, pH control at 6.8-7.0. After 30 hours of fermentation, the measured dry cell weight was 98g / L.
[0041] Take 200ml of fermentation broth, centrifuge the bacteria, add 30ml of water, adjust the pH to 12 with 30% sodium hydroxide, heat to 90°C, keep for 35 minutes, vacuum filter, and dry the bacteria at 80°C. Put the dried cells into a reflux bottle, add 100ml of acetone, heat and reflux for extraction for 30 minutes, vacuum filter, and evaporate the filtrate to dryness. Add 50ml petroleum ether into the container, after dissolving the oily substanc...
Embodiment 2
[0043] According to the process described in Example 1, Alcaligenes eutropha H16.65-7 was fermented for 33 hours, and the measured dry cell weight was 105 g / L.
[0044] Take 500ml of fermentation broth, centrifuge the bacteria, add 100ml of water, adjust the pH to 11 with 30% ammonia water, heat to 95°C, keep for 40 minutes, centrifuge, and dry the bacteria at 75°C. Put the dried cells into a reflux bottle, add 200ml of acetone, heat and reflux for extraction for 40 minutes, vacuum filter, and evaporate the filtrate to dryness. Add 150ml of petroleum ether into the container, dissolve the oil, evaporate the petroleum ether to about 50ml, and separate by silica gel column chromatography. Collect the yellow liquid, evaporate the petroleum ether to dryness, add 50ml of absolute alcohol, and heat to 45°C to completely dissolve the oily substance. Take a sample for testing, and the sample shows a unique blue reaction of coenzyme Q10. Thereafter, the samples were placed in a refrig...
Embodiment 3
[0046] According to the process described in Example 1, Alcaligenes eutropha H16.65-7 was fermented for 36 hours, and the measured dry cell weight was 125 g / L.
[0047] Take 1000ml of fermentation broth, centrifuge to separate the bacteria, add 200ml of water, adjust the pH to 12 with 30% potassium hydroxide, heat to 90°C, keep for 60 minutes, centrifuge, and dry the bacteria at 75°C. Put the dried cells into a reflux bottle, add 400ml of acetone, heat and reflux for extraction for 40 minutes, vacuum filter, and evaporate the filtrate to dryness. Add 300ml of petroleum ether into the container, dissolve the oil, evaporate the petroleum ether to about 50ml, and separate by silica gel column chromatography. Collect the yellow liquid, evaporate the petroleum ether to dryness, add 100ml of absolute alcohol, and heat to 45°C to completely dissolve the oily substance. Take a sample for testing, and the sample shows a unique blue reaction of coenzyme Q10. Thereafter, the samples wer...
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