Operating instruction for BIOXING bioreactorThe components for the bioreactor1-variable area flowmeter:used for regulate the flow rate of air2-the bioreactor: the place for cultivating strains and fermentation3-the base:for supporting the bioreactor and alkali bottle4-feeding pipe and alkali(acid) pipe: for added medium and alkali or acid5-condenser: used for cooling the out air6-air outlet: for air exhausting7-rotator: install the electrical machine for driving the rotator8-inoculate: adding strain, medium, glucose and other needed for fermentation9-use for installing pH electrode10-used for installing oxygen electrode11-used for installing temperature electrode12-used for installing the pipes for water used for keeping the temperature at a set point13-sampling: used for taking samples from bioreactor14-nuts: for fix the lid and make sure the bioreactor is gas tightness 15-air inlet: for pumping aseptic air to bioreactor16-parameters sleet17-user loading in(or out)18-electrical machine: for driving the rotator19-temperature electrode20-bioreactor switchThe reactor(contain at least 1.5L water in it) must be sterilized at 121℃for 30min before used for fermentation, after this, the reactor will be able to use for fermentation.Instructions for bioreactor use1.Turn on the power and login the systemUser name: SYS Password: 3332.After loading successfully, some necessary steps must be done before thefermentation, this may conclude the followings.A.Electrode calibration: the electrodes used for fermentation are pH and oxygenelectrode in our lab and the calibration for them is similar.pH: use the standard buffer(7.0,green and 4.0,pink) to calibrate the electrode: pH 7.0 is first and then 4.0Oxygen: use the saturated sodium sulfite calibrate the zero point and before inoculate the 100% point must be calibrateB.Feeding pump calibration: as the process of fermentation is continued, extrafresh and aseptic fermentation medium may be added under a constant rate, so the feeding pump must be calibration as you calculated.3.Bioreactor wrapping and fermentation mediumThe fermentation medium is added to bioreactor and sterilized at 121℃for 15min.Before this, following things must be done.A.Bioreactor assembling: after all the things above are done, first we must assemblethe bioreactor, there may have some points need to be considered. First the seal ring under the lid must be added correctly and four nuts must be screwed. Another one is the direction of the condenser, it must be placed different side compared with the scale in the glass.B.Bioreactor wrapping: in order to ensure the aseptic environment for fermentation,the wrapping is necessary: first, the pipes for taking samples, air output and medium feeding must be wrapped by silver paper or gauze (eight layer); and then two air filter must be added at the entrance of pipe for air input (notice the direction, the inlet is faced outside) and then wrapped by silver paper.C.Fermentation medium preparation: weighing all the chemical reagents anddissolved by tap water. In this step, the water added for regents dissolving is notthe same as the final volume(1.5L),the water volume is calculated by the equation as followed:The volume of water=1.5-V1-V2V1:the volume of glucose or other carbon source(the carbon source is sterilized at 121℃for 15min separately)V2: the volume of strain for fermentationD.The electrode must be inset in the appropriate place(all the electrode used forcontrol the conditions of fermentation and must be calibration before used)4.operation of bioreactor after fermentation1)In order to keep the aseptic environment, a positive pressure must be added inthe bioreactor, so the first step after sterilized is pouring the aseptic air(or carbon dioxide) into bioreactor immediately, and then starting stir at 150rpm2)Insert the temperature electrode into the bioreactor and install the condensatewater pipes, then open the inlet and outlet valve and convert the temperate controller from manual operation to automatic. At the same time, install the pH and oxygen electrode, the alkali or acid bottle and the feeding bottle.3)After the temperature decreased to the fermentation temperature, convert thepH controller from manual operation to automatic and regulate the pH to set point.4)For the oxygen electrode, calibrate the 100% point when the pH stabilizes atthe set point, this operation is only suitable for aerobic fermentation, when under anaerobic condition, the oxygen electrode isn’t calibrated but it is necessary for air impermeability of the bioreactor.5)After all steps above are OK, the strain and carbon source will be inoculated.In this step, the necessary regent added to bioreactor is different from strain to strain, and the regents contain VB1, biotin and antibiotics.6)Along the whole fermentation, we need to take samples from bioreactor foranalysis some parameters, such as OD and residual glucose. When take the sample, the pipe for air out must be locked.7)When the fermentation comes to an end, sodium hydroxide must be added tobroth for killing all the strain and then poured the waste broth to sewer and the bioreactor must be cleaned.。