 Let us start from the lab scale fermenter, first I will give you some brief introduction about the fermenter which we will use in our experiment. So, we will use this 7.5 liter glass vessel having the round bottom, top plate is a stainless steel of New Brunswick company. The model of this fermenter is BioFlow 110. So, you can see the whole assembly of this fermenter, this part is the, this part is known as the processor and this part is for the dissolved oxygen probe and pH probe and this part is for Watton and Wattoff and this is for the control of the temperature and the sensing of the temperature and other is the supply of the electricity to the main motor of this fermenter. As comes in the last part, these having the four different pumps which is worked as a parallelistic pump which will be used to feed anything either that is a acid or a base or anti foam or some medium into this vessel. So, first of all along with this main assembly of the fermenter as you know the basic part of the fermenters are this one is body vessel, side is an agitation mechanism along with this motor and then also along with that is some impellers and then is the sparser. So, there are some accessories along with this fermenter. The first accessory is that is the chiller that will use to provide the chilled water to this condenser just to check the evaporation loss and other accessory of this is an autoclave because this fermenter is a lab scale fermenter and we have to sterilize this vessel exetive. So, we will use this floor model autoclave for the sterilization as an accessory of the fermenter. The other important accessory for this fermenter on my left side that is the compressor that will give us a compressed air in order to maintain the air pressure inside that. So, just keep in mind and remember your module in which we were talking about the differentiation about the lab scale fermenter, pilot scale fermenter and industrial scale fermenter. In this laboratory we have two fermenters this side is a lab scale fermenter 7.5 liter is maximum capacity and the working volume if we use the 75 percent of the total volume of this then there will be 5 liter will be the working volume. So, you can see here the pilot scale fermenter almost the assembly is same, but in this stainless steel new brevins model is 4500 which we called as pilot scale fermenter because this fermenter has in-stew sterilization process. You can see on other side is a automatic control of the steam which utilize to sterilize to in order to maintain the same temperature which require for the sterilization. So, but we will focus only the laboratory scale fermenter that well we will use in our current experiment. So, if we go in detail one by one. So, first of all before to start on that we have to de-assemble this whole fermenter assembly. So, these we have to remove all the gadgets one by one later on I will told you all these gadgets. So, be careful when we are de-assembling all these gadgets. So, this is a temperature probe we should be very care should we take a very care full attitude while handling this. So, now we have separate this body vessel. So, we will just open the top plate I will now request to me to help me. So, you can see that this is the top plate along with the you can see here this is the shaft having 2 impellers and these are the Russian type impellers having 6 blades. So, this is a proper seal and this is the condenser and this is the jacket which we can use a coil for cooling and heating of this and this is for the temperature probe and this is a dissolved oxygen probe. So, the main part of this fermenter which we called as the body vessel. So, this is a borosilicate glass vessel having the capacity of 7.5 liter capacity. So, this is basically the stand in which that hold this body vessel. So, first of all we will through all the material and then we have a wash of this vessel. So, after this after washing we will add our medium which we to utilize for the production of the form is. As concerned the media formulation we have used we will use 2 percent raw storage and 0.5 percent potassium dihydrogen phosphate and 0.5 gram per liter ammonium sodium nitrate and 0.2 gram per liter magnesium sulfate and 0.1 gram of calcium chloride. We have already made this medium. So, we will add this medium into this vessel thoroughly mixed it. So, we added here the 5 liter medium into this flask. So, then now we will reassemble this fermenter, but the main precautions in the reassembling we have to be very careful in observing this seal because this seal should be properly fixed because if the seal is not properly fixed then there is a chance of contamination through. So, now it is ok. So, I will show you this baffle. In this glass body vessel fermenter the baffle part is the separate. So, we will fix along with it. So, after fixing this we will close this top with this because during the sterilization the seal will not be affected just for increasing the life of this. And then we have to be check all the gases should be properly closed and then we have to cover this and then we have to fix this. And then we have to fix ascent. You can see in one bottle we have added the anti foam it is a synthetic anti foam. Another bottle we have 5 normal NOH and then we have a 5 normal acetic acid. And in other bottle we have a just a water to make the containment level better. So, you can see here on the top plate there are the 3 needles. So, we will gradually collect all these 3 tubes. So, one for acid, one for alkali and one for anti foam. These are the 3 different just for the safety. We will first close because when we have to sterilize sometimes due to the back pressure and if there is a positive pressure inside then acid base and the anti foam can be added into the bottle. The body vessel and disturb the whole medium. So, these are the 3 point port and we attach these and you see these are the 3 filters. Point 2 micron size is the filter size because when there is a session and then there will be a pressure. So, the air should be go inside and they remain sterilized. So, this water we will just attach on the top of the condenser because sometimes when there is an electric shutdown and there will be a negative pressure inside what happen the air can be comes from the outside to inside. If we add the air filter here then there is a bright chance of the choking of that filter due to the wet air. So, sometimes so as my experiences we mostly attach this. So, it will be sterilized along with this whole assembly inside that. So, we will put this whole assembly into this autoclave before all this there is one more part the air sterilization system. So, you know there is a compressor that will give us the compressed air and after passing through this rotometer it will go to first coarse filter. This coarse filter will first separate the dust particles and the moisture from the air. Then it will go to the second filter which we called as the fine filter it ensure the aseptic entry of the air into the body vessel. So, this third filter we used for the more safety sometime it happen when there is a back pressure. So, then this pressure can be choked. So, this is the safety filter which we attached in the series of this. So, here you can see that we will attach this, but we will attach a blocker here we will make a small cotton plug here. When this whole assembly is ready then we put this into the autoclave, but before putting it into the autoclave we have to lose this one plug because sometime during the sterilization process there is a high pressure because the media will expand just for the safety of this we have just a lose because due to the extra pressure the vessel cannot be ruptured or a blast. So, we will put it into the fermenter into the autoclave and we will. So, you can see that we have put all the accessories of acid, base and anti foam and with the air sterilization system along with the body vessel inside the autoclave. So, we will just close it. So, now it is almost close now we will it is ready for the autoclave. So, now we will own it and it almost take 40 minute to complete this whole autoclave process. So, after 45 minute we will proceed the next. So, as I have already told you that autoclave will take 40 to 45 minute. So, it is after 45 minute I think it is over now you can see that the pressure on the autoclave is 0 degree. So, before open this we have to ensure that this needle of the pressure should be at 0. So, if there is inside the pressure we should not open this the lid. So, now it is ok. So, we can open it. So, it is somehow hot. So, before taking the whole material out we should be very careful because the whole material inside will be very hot. So, we should use the gloves cotton gloves. So, we will gradually take these, but before take we have to be close the valve which we have the lid and then we will take the whole material on the side. Help me by putting please Muhammad Kaussan comma. So, two and three gadgets we have to take it out. So, as you see that we have put back the whole assembly here on the shelf after the autoclave. So, now we will fix the whole gadgets back. So, these are the different strands three strands one for acid one for alkali and one for antifol. First of all we will fix the air filtration system. Here is the stand and we will fix this pipe air pipe that comes from the compressor and then this rotometer to this. So, first of all as you see that we fix the air circulator system from starting from the compressor to the rotometer and then to this coarse filter and then finally to this. So, then we will fix the motor by removing the cap we will just see these two cut and we will fix it like this and then pH sensor we will put here and then dissolved oxygen probe and then we will fix and then we will fix the wet town and wet off. As concerned the wet town and wet off we have this and then is the sensor the same way here and then this and then is a temperature probe. It is a P T 100 probe and we will add it here you can see here. So, these are the three one is acid one is base five normal and OH and then is antifol which will use as we needed. So, you can see now we have reassembled the whole, but one thing is remaining that is the use of the attachment of the chiller with this. These are the pipe you can see here there are the two pipes one the out from the chiller that will be the in of the chiller on this side and then we also use on the circulating on this side. The in of the chiller will be the out of this. So, almost we have completed the whole assembly. So, it is the whole assembly of the firm we have reassembled. So, now first of all we will make sure the start of the air feeding in suicide it and then we start feeding the chilled water and for controlling the temperature we have to attach the jacket of the fermenter. So, it means the jacket of this fermenter. So, this fermenter has this blanket type heat jacket. So, we will fix it. So, now we will fix it here. So, now this fermenter is ready for further process which is inoculation, but before inoculation we will on this whole fermenter. You can see this is a new Brunswick bio flow 110. It will takes a moment to start on. You can see the saccharine of that the temperature at this time is 24.8 and the set point is 30 degree Celsius and the rpm we can fix it. Set point we will fix it 200 rpm. So, you can see here now the agitator is now on pump A pump B and pump C at this time the pH is 6, but we need a set point is 5.5. So, we can fix at 5.5. So, dissolved oxygen at this time is 0.1 because after the heating there will be less. So, now we will on the compressor. Now, the compressor is on. It will automatically off when it will keep its pressure. So, come on here. Before on this we will open the blockage from the here. Now, you can see the spudging in the air and then mixing. Now the fermenter is ready for inoculation. Let us start inoculation. As you know that in our previous step before making the ready the fermenter we have inoculate 6 different flasks with the cunidial inoculum and we put them into the shaking incubator. Now we will remiss is already having those 6 flasks. So, we have these 6 different flasks of the inoculum. You can see here the inoculum which you have already seen there. So, we need a cotton. But before starting the inoculation process first we have to increase the air pressure and then we have to lose a little bit of this lead which we called as the inoculation lead. We have to first matches. We have to just measure it out with the alcohol. So, this step should be very critical and we have to be very careful because there are 2 alternative methods for inoculation. So, first we have to be fire just to sterilize this area. You can see that it is now automatically be sterilized and this we will create the aseptic environment. So, we will gradually add first inocule we will open this in this zone and then we will add. By this way we will add all the 6 into the vessel. When we completed this we have to reclose this. So, we have to then we have a normally course when we will dip in the water then we can gradually do this. But just for more we can make it with this. So, now it is you can see here inside you can see inside the inoculum beads. So, after the inoculation we will give the proper incubation time and gradually we will take out the samples. So, for the sampling we have to take the material gradually at 0 time. At the start of the fermentation at 0 time we will take the sample. For the sampling we can use a syringe. First of all we will fix this on this and then we will remove this blockage and then we first part we will have to be give me a one flask or a beaker. First 5 ml we will discard and then because that. So, that is the sample which we can say that the sample as 0 time. So, on each time when we will take out the sample we will calculate the biomass produced and the carbon source starch utilized and then the product which we called as a mileage production and the protein formed. So, at each step of the sampling which we will take after our 4 hours interval. So, on each interval we will check as I have already told you that biomass production, substrate utilization and product formation in order to calculate the exact kinetics of this fermenter.