Method for producing reduced coenzyme Q10
A production method and reduction technology, applied in the field of biological production and coenzyme Q10 production, can solve the problems of molecular oxidation, environmental pollution of reduced coenzyme Q10, and harmful effects of coenzyme Q10
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Embodiment 1
[0026] The human liver cell line Fa2N-4 (ATCC-5566) was selected as the parent material for the production of reduced coenzyme Q10.
[0027] From DMEM-1 ((high glucose) containing 4500mg / L D-glucose, L-glutamine and 25MmHEPES without sodium pyruvate and sodium bicarbonate) culture solution pH7.4, in 2.2L, DMEM-1 culture solution 3L cell generator, sterilized at 121°C for 10 minutes, add the mother material for the production of reduced coenzyme Q10-hepatic cell line Fa2N-4 (ATCC-5566) accounting for 10% of the volume of the culture medium, at 37°C, 5% Cultivate for 48 hours under the conditions of CO2 and 300 rpm stirring speed.
[0028] At the end of the cultivation, 30ml of 25% citric acid solution was added to the culture medium, and the 180g cell-containing wet body obtained by centrifuging the culture was collected and washed with water for 3 times to obtain a clean cell body.
[0029] Then, use a Mulitibeads shocker (multi-cell culture shaker), B300 to shake, stir for 1...
Embodiment 2
[0038] Human serum albumin was selected as the parent material for the production of reduced coenzyme Q1. From DMEM-1 ((high glucose) containing 4500mg / L D-glucose, L-glutamine and 2525Mm HEPES without sodium pyruvate and sodium bicarbonate) culture solution pH7.4, in 2.2L, DMEM-1 culture solution The 3L cell generator was sterilized at 121°C for 10 minutes, and 10% of the volume of the culture medium was added to the cell parent material for the production of reduced coenzyme Q10—human serum albumin. Cultivate for 48 hours at 37° C., 5% CO 2 , and a stirring rotation speed of 300 rpm.
[0039] At the end of the cultivation, 30ml of 25% citric acid solution was added to the culture medium, and the 180g cell-containing wet body obtained by centrifuging the culture was collected and washed with water for 3 times to obtain a clean cell body.
[0040]Then, use a Mulitibeads shocker (multi-cell culture shaker), B300 to shake, stir for 1 hour, break the cell body, and centrifuge to...
Embodiment 3
[0052] Chinese hamster ovary cells, CHO cells, were selected as the parent material for the production of reduced coenzyme Q10. From DMEM-1 ((high glucose) containing 4500mg / L D-glucose, L-glutamine and 25Mm HEPES without sodium pyruvate and sodium bicarbonate) culture solution pH7.4, in 2.2L, DMEM-1 culture solution The 3L cell generator was sterilized at 121°C for 10 minutes, and 10% of the volume of the culture medium was added to the parent material for the production of reduced coenzyme Q10—Chinese hamster ovary cell CHO cells. Cultivate for 48 hours at 37° C., 5% CO 2 , and a stirring rotation speed of 300 rpm.
[0053] At the end of the cultivation, 30ml of 25% citric acid solution was added to the culture medium, and the 180g cell-containing wet body obtained by centrifuging the culture was collected and washed with water for 3 times to obtain a clean cell body.
[0054] Then, use a Mulitibeads shocker (multi-cell culture shaker), B300 to shake, stir for 1 hour, break...
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