High-density culture method of prorocentrum
A technology of high-density culture and Prototheca, applied in microorganism-based methods, biochemical equipment and methods, single-cell algae, etc., can solve the problems of high price, small capacity of light incubator, and high energy consumption of temperature control system, To achieve the effect of simple operation, efficient acquisition, and small culture volume
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Embodiment 1
[0024] In a sterile operating room, suck 19.0L of sterilized f2 medium with a salinity of 28 into a 20L jacketed glass reactor with a sterile silicone tube, and inoculate a concentration of 5.0×10 at a volume ratio of 5% 6 cells / L Prorocentrum PM03, that is, add 1.0L of algae seed solution. Transfer the reaction kettle to a conventional laboratory, use an air pump to connect to a sterile 0.2μm filter and a silica gel tube to vent 3L / min, the speed of the reaction kettle is 80rpm / min, the light is 5000lux, the light / dark cycle is 14h / 10h, and the jacket circulating water temperature Control 22-25°C and cultivate for 15 days. Then in the sterile operating room, most of the cultured algae liquid is discharged from the reactor outlet, and a certain proportion of the volume is reserved for inoculation in the next culture cycle. Using the Sedgewick-Rafter plate to count the cells under the microscope, the cell density of the cultured algae solution was 8.97×10 7 cells / L.
Embodiment 2
[0026] Continuation of Example 1. In a sterile operating room, the cell density retained in a 20L jacketed glass reactor is 8.97×10 7 1.5L of Prorocentrum PM03 seed solution in cells / L, inhaled with a sterilized silicone tube and added 18.5L of sterilized f2 medium with a salinity of 25. Transfer the reaction kettle to a conventional laboratory, use an air pump to connect to a sterile 0.2μm filter and a silica gel tube to vent 3L / min, the speed of the reaction kettle is 80rpm / min, the light is 4000lux, the light / dark cycle is 14h / 10h, and the jacket circulating water temperature Control 22-25°C and cultivate for 15 days. Then in the sterile operating room, most of the cultured algae liquid is discharged from the reactor outlet, and a certain proportion of the volume is reserved for inoculation in the next culture cycle. Using the Sedgewick-Rafter plate to count the cells under the microscope, the cell density of the cultured algae solution was 1.38×10 8 cells / L.
Embodiment 3
[0028] Continuation of Example 2, in a sterile operating room, the cell density retained in a 20L jacketed glass reactor is 1.38×10 8 Cells / L of Prorocentrum PM03 seed solution was 2.0L, and 18.0L of sterilized f2 medium with a salinity of 25 was added to the sterilized silicone tube. Transfer the reaction kettle to a conventional laboratory, use an air pump to connect to a sterile 0.2μm filter and a silicone tube to vent 3L / min, the speed of the reaction kettle is 80rpm / min, the light is 3000lux, the light / dark cycle is 14h / 10h, and the jacket circulating water temperature Control 22-25°C and cultivate for 15 days. Then in a sterile operating room, discharge most of the cultured algae solution 18L from the reactor outlet, and use the Sedgewick-Rafter plate to count the cells under the microscope to find that the cell density of the cultured algae solution is 2.05×10 8 cells / L; The remaining 2.0L of algae seed solution is used for inoculation in the next culture cycle, add 18L o...
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