1、 Sterilization operation
1. Close all water and air supply pipelines. Open the steam pipeline valve. At the same time, slightly open the exhaust valve of the fermentation tank jacket to discharge the remaining water from the jacket.
2. Turn on the mixing motor of the fermentation tank and set the speed to 200rpm to evenly heat the fermentation broth. When the temperature rises above 95 ℃, stirring can be stopped. Then wait for the temperature to rise to 121 ℃ (tank pressure at 0.1-0.12Mpa) to start timing.
After the timer starts, the fermentation time is usually 20-30 minutes. During this time, the temperature should be ensured to be no lower than 120 ℃ At the same time, sterilization of air filters and air pipelines can be carried out.
4. Sterilization of air filters and air pipelines: Slightly open the drain valve of the filter and the diaphragm valve of the air pipeline to ensure steam sterilization of the air pipeline. But it cannot be opened too large to prevent a large amount of steam from entering the tank and diluting the culture medium.
5. Slightly open the steam valve and outlet valve of the discharge and sampling valves to ensure sterilization of the pipeline. The inoculation port on the lid of the fermentation tank also needs to be deflated to meet sterilization requirements.
When the insulation is completed, the diaphragm valve in the air pipeline should be closed first. Close the drain valve of the air filter, as well as the outlet valve and inoculation cap of the sampling valve. Then close all steam valves.
7. Open the cooling water valve and drainage valve, as well as the air flow meter and air vent valve, and blow dry the air filter. At this point, it is necessary to pay attention to changes in tank pressure. The tank pressure cannot be lower than 0.02Mpa. When the tank pressure reaches 0.05Mpa, immediately open the air pipeline to ensure that the fermentation tank pressure is around 0.05Mpa.
When the temperature drops to 95 ℃, the stirring can be turned on. When the temperature drops below 50 ℃, the automatic temperature control mode can be activated to reach the inoculation temperature of the culture medium, and the sterilization process is completed.
2、 Operation of fermentation process
1. Vaccination: The inoculation method can be flame inoculation or differential pressure inoculation.
(1) Flame inoculation method: disinfect the inoculation port with an alcohol ring, then open the inoculation port cover, quickly pour the inoculation solution into the can, and tighten the cover.
(2) Differential pressure method: Place a gasket before sterilization, open the inoculation port cover during inoculation, and first pour in a certain amount of alcohol for disinfection. After a moment, insert the needle of the seed bottle into the gasket of the vaccination port. Using the pressure difference between the tank and the seed liquid bottle, introduce the seed liquid into the tank and tighten the lid.
2. Tank pressure
During the fermentation process, it is necessary to manually control the tank pressure by using an outlet valve to control the pressure inside the tank. When adjusting the air flow rate, the outlet valve must be adjusted at the same time to maintain a constant pressure in the tank greater than 0.03Mpa.
3. Measurement and control of dissolved oxygen (DO)
(1) Calibration of dissolved oxygen: Before inoculation, at a constant fermentation temperature, the dissolved oxygen DO value at the maximum value of the rotation speed and air volume is taken as 100%.
(2) Measurement and control of dissolved oxygen (DO) during fermentation process: DO control can be achieved by adjusting air flow rate and speed. The simplest is the correlation control between rotational speed and dissolved oxygen. Secondly, it is necessary to simultaneously adjust the intake volume (manually) for control. Sometimes pure oxygen needs to be introduced (such as in high-density cultivation of certain genetically engineered bacteria) to achieve the required DO value.
4. Measurement and Control of pH
(1) PH calibration: The pH electrode should be calibrated for pH before sterilization.
(2) The control of pH value during fermentation is achieved by adding acid and alkali using a peristaltic pump, and the acid or alkali bottles must be sterilized in a sterilization pot first.

3、 Operation of the controller
1. Controller startup: Turn on the power, first press the "S/E" key on the membrane keyboard, then press the "confirm" key to start the fermentation control program; At this point, if the water in the heater is not fully filled, the program will automatically perform the water inlet operation; After the water is filled up, users can control various actuators according to the operation method of the lower computer controller mentioned above.
2. Controller operation: Use the F1-F6 keys to switch the interface on the LCD screen to the interface that the user needs to control. Use the directional keys to move the cursor on the interface to the variable that needs to be controlled. If you want to change the operating mode, simply press the confirm key. If you need to enter numbers on the keyboard, press the confirm key after entering the numbers. For example:
(1) Temperature control: In manual mode, manual operation of temperature is relatively simple, just change the control quantity in manual mode. Enter the temperature control interface by selecting the shortcut keys (F1-F5), then move the cursor to point to "manual mode", and press the "confirm" button to enter the manual mode of temperature control. At this point, a small hand will appear after "manual mode" to indicate that the current selection is manual mode. Move the cursor to the manual setting area and select "Control Quantity" by moving the cursor up and down. Enter the desired control output value by pressing the number keys, such as 80, and press the "confirm" button to confirm; (Note: The control range is 0-100. When the input control value is greater than 50, it is in the heating state, otherwise it is in the cooling state.).
(2) Speed control: Use the cursor movement key to move the cursor to the "set value" position. Enter 300 on the numeric keypad, and the value of "300" should appear after the "set value"; Then press the 'confirm' button to confirm the input. If there is an error in the input, you can press the "Clear" button to clear the data.
4、 Operation of steam generator
1. Turn on the water inlet switch to raise the water level in the storage tank to the highest level and maintain the water inlet state
2. Connect the generator power supply and supply water to the pot to the normal water level (50-80% of the liquid level pipe), which should not exceed the highest water level and should not be lower than the lowest water level. Close the steam outlet valve
3. Plug in the power, turn on the power switch, the work indicator light will turn on, and start heating the pot water
4. Connect the steam pipe to the interlayer pipeline system of the fermentation tank, open the relevant valves, keep the pipeline unobstructed, and close the exhaust valve of the fermentation tank at the same time. When the pressure rises to the working pressure (2kg/cm2), open the steam outlet valve to supply steam
5. After use, turn off the power first, then turn off the inlet pipe. After the generator cools down appropriately, drain the sewage from the pot.
【 Precautions 】
The set pressure of the safety valve has been adjusted by the manufacturer and cannot be adjusted arbitrarily. If the safety valve is found to be malfunctioning, a new one should be replaced. It is strictly prohibited to change the automatic pressure control function and parameters without authorization.
During normal operation, discharge at least once every 8 hours and promptly use sandpaper to remove scale from the water level probe
3. During use, it is strictly prohibited to close the safety valve, modify or block it with a plug without authorization
4. The water storage bend of the pressure gauge should be removed and cleaned regularly

5、 Operation of air compressor
1. Plug in the air compressor power supply, turn on the air compressor, and start the machine for 15 minutes in a no-load state.
2. If there is no abnormal noise after starting, close the air outlet and connect the air compressor outlet pipe to the air purifier, parallel to the fermentation tank air inlet pipeline.
3. When the air pressure rises to 2kg/cm2, open the gas supply valve and turn on the air switch to provide sterile air to the sterilized fermentation tank. After the air pressure reaches the set pressure, the pressure switch automatically cuts off the power and the motor stops running.
4. The operating pressure of the air compressor shall not exceed the rated working pressure. If adjustment is required, it must be carried out by a dedicated salesperson and cannot be adjusted by oneself.