Method for producing Dendrobium huoshanense alkaloids by cell suspension culture
A technology of suspension culture cells, Dendrobium Huoshanense, applied in biochemical equipment and methods, microorganisms, plant cells, etc., can solve the problems of low yield of target products, low growth rate of biomass, long production cycle, etc., and shorten cell culture cycle, uniform supply, and the effect of improving product yield
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Embodiment 1
[0024] This embodiment discloses a method for producing Dendrobium alkaloids by suspension culture cells, comprising the following steps:
[0025] 1. Training stage
[0026] (1) Single cell preparation
[0027] The capsules of Dendrobium Huoshanense were taken out, sterilized according to the requirements of tissue culture technology, and then aseptically sowed. The sowing medium was a solid 1 / 2MS medium. After the protocorms germinated, they were taken out, and the protocorms with bright color, large individuals and vigorous vitality were selected. Separation and preparation of single cells by enzymatic hydrolysis, filtration with a microporous filter with a pore size of 80 microns, separation of the filtrate by a low-speed centrifuge, and collection of precipitated cells;
[0028] (2) Single cell line culture
[0029] Inoculate the collected single cells into a baffle shake flask containing the first MS liquid medium, the inoculum amount of single cells is 23g / L, the light...
Embodiment 2
[0041] 1. Training stage
[0042] (1) Single cell preparation
[0043] The capsules of Dendrobium Huoshanense were taken out, sterilized according to the requirements of tissue culture technology, and then aseptically sowed. The sowing medium was a solid 1 / 2MS medium. After the protocorms germinated, they were taken out, and the protocorms with bright color, large individuals and vigorous vitality were selected. Separation and preparation of single cells by enzymatic hydrolysis, filtration with a microporous filter with a pore size of 80 microns, separation of the filtrate by a low-speed centrifuge, and collection of precipitated cells;
[0044] (2) Single cell line culture
[0045] Inoculate the collected single cells into a baffle shake flask containing the first MS liquid medium, the inoculum amount of single cells is 25g / L, the light intensity is 3000lux, the color temperature is 5000K, shake culture, adjust the pH value to 5.8, and the temperature At 22°C, when the cell...
Embodiment 3
[0055] 1. Training stage
[0056] (1) Single cell preparation
[0057] The capsules of Dendrobium Huoshanense were taken out, sterilized according to the requirements of tissue culture technology, and then aseptically sowed. The sowing medium was a solid 1 / 2MS medium. After the protocorms germinated, they were taken out, and the protocorms with bright color, large individuals and vigorous vitality were selected. Separation and preparation of single cells by enzymatic hydrolysis, filtration with a microporous filter with a pore size of 80 microns, separation of the filtrate by a low-speed centrifuge, and collection of precipitated cells;
[0058] (2) Single cell line culture
[0059] Inoculate the collected single cells into a baffle shake flask containing the first MS liquid medium, the inoculum amount of single cells is 27g / L, the light intensity is 4000lux, the color temperature is 6000K, shake culture, adjust the pH value to 6.2, and the temperature At 24°C, when the cell...
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