Shake flask cultur is in the stage of strain screening and culture (pilot test), the culture conditions should be as close as possible to the fermentation production culture conditions, the workload is large, long time, complex operation. The factors that affect the high efficiency of shaking flask culture are mainly culture temperature, shaking amplitude of the shaker, the amount of shaking flask, the pH of the culture medium, the viscosity of the medium, etc. Culture temperature: the mycelium growth temperature of different edible fungi is also different, most of the suitable growth temperature is between 22 ℃ and 30℃, if the culture temperature is too low, the mycelium growth is slow; When the temperature was too high, the mycelium pellets were loose and sparse, and the vitality and quality of mycelium pellets decreased.

Shaking frequency and high efficiency shaking bottle loading: edible fungi are aerobic fungi, liquid culture, mainly by absorption of oxygen dissolved in the culture medium. The dissolved oxygen in the culture medium is mainly affected by the viscosity of the medium, the amount of liquid in the container, the frequency of oscillation and other factors. The shaking frequency is large, the shaking flask is small, the concentration of the medium is up to, the dissolved oxygen of the medium is high, and the other way around is low. Usually rotary shaker speed is 180-220 RPM /min, reciprocating is 80-120 RPM /min, amplitude 6-7cm.

Ph of culture medium: PH of culture medium directly affects nutrient absorption, enzyme activity and mycelial pellet growth. Specific pH must be adjusted before sterilization, most edible fungi in pH 2.0-6.0. In order to prevent the drastic change of PH in the culture medium, calcium carbonate, phosphate and other buffer substances are often added to the culture medium.

Medium viscosity: Medium viscosity directly affects the amount of oxygen dissolved in it, and also affects the formation of mycelial pellets. The results showed that when the viscosity of culture medium increased, the diameter of mycelium pellets decreased, the number increased, and the yield increased. Therefore, according to the requirements of liquid strains on the diameter of mycelium pellets, the culture medium with a certain viscosity should be configured. Cell culture is a rigorous work, especially when it needs to be cultured with the help of a shaker, such as high-efficiency shaker, it should be considered more comprehensively, so as to ensure the smooth progress of cell culture.