DAY 2 - PREPARATION OF OUR BIOREACTOR

EPISODE 2,  GET TO KNOW YOUR BIOREACTOR  DEEPLY!!

Date:    5.10.2010 (Tuesday)

Time:    8.00A.M

Place:   Room 148 PPTI 

After reaching the lab, we prepared seed culture, we using sterile toothpicks to scrape up a single colony aseptically. The work will be completed by transferring the colony into the shake flask which contained medium. We left it overnight (18-24 hours) in the incubator shaker.

  Colonies of S.cerevisiae

Today is to set up the bioreactor. Mr. Shaman showing to us step by step how functioning a bioreactor. First, he prepared the buffer pH 4.01 and buffer pH 7.01. Then he took the pH probe and removed the cap on the bottom and scraped platinum to make sure it is neutral. He put the probe in the pH buffer to set the point of the calibration curve, C1 is pH 7.01 and C2 is pH 4.01. When we calibrated, need to insert the pH probe together with the temperature probe to allow comparison.

Both pictures show the way to calibrate the pH probe

Then, he poured the medium into the vessel with the adding of 15ml distilled water to make sure that no evaporation lost during autoclaving the medium.

The medium was poured into the vessel

  

He took the pO2 electrode and removed the green plastic end cap. Then, the bottom metal section was unscrewed to check whether there was liquid electrolyte in the membrane cartridge inside. There is a bottle provided from the company to top up the liquid electrolyte if the level of the liquid is low. After that, he replaced the end section, and fit it into the vessel vertically and secured it.

Then he prepared the reagent which is antifoam liquid by adding around 10ml in the 250ml bottle. Make sure a silicone dip tube is connected to one metal pipe on the head plate. The tubing between the bottle and the pump head is clamped off.

Then he set up other parts of the fermentor, exit gas cooler with filter, inlet filter connected to sparger, free port for inoculation and sample device. Need to double check all these accessories already completely set up. Then need to use aluminium foil to close all the parts that will direct exposed to the air before autoclaved, so that the fermentor will remain sterile after taking out from the autoclave.

Bioreactor is ready for the autoclaving

Bioreactor in the autoclave

Bioreactor after the autoclaving

After the bioreactor was autoclaved, it needs to be polarized with oxygen for about 2 hours... so for this day is end at this state and will proceed tomorrow with innoculation and sampling procedure.