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Wednesday, July 16, 2008

40 L Fermenter

Ø Sampling
1. 2 hrs subsequent to inoculum transfer, the sterile bottle was clamped at the sampling port and the sampling line was steam sterilized for 10minutes.
1. Sample was collected by first draining off a few ml media.
2. pH and the O.D. (at 600nm) were checked and the specific growth rate (µ) was calculated.
Ø Dosing
1. Peristaltic pump was calibrated.
2. 400 ml of autoclaved Dextrose solution was transferred to a sterile transfer bottle in laminar airflow.
3. The 400 ml was transferred to the vessel in equal amounts in 4 hours at six-pulses/ hour.
4. 16.66 ml of the solution was transferred to the vessel at a ten-minute interval over the 4 hours.
5. A sample of the media was taken every one hour.
6. OD was checked at 600nm and the specific growth rate (µ) was calculated.
Ø Harvesting & CIP
1. On the third day of fermentation, the fermentation process was shut down and the biomass was harvested into the harvest vessel by pressurizing the vessel.
2. The dosing bottle of dextrose and antifoam were unclamped.
3. CIP skid was connected to the fermenter.
4. 9 liters (30% of working volume) of soft water was filled in the mixing vessel in the C.I.P station.
5. Appropriate valves were opened such that water entered the fermenter vessel through the spray ball and was agitated for 5 minutes. Water was then drained.
6. Washing with 1% alkali (NaOH) was done
- Concentrated alkali was prepared in the alkali bottle in the CIP station
- 1 liter of this concentrated alkali was pumped into the vessel and mixed with 8 liters of water and was heated up to 50°C and supplied to the fermenter vessel through the spray ball.
- The spray ball supply was stopped and alkali supplied to the transfer lines, one at a time for 5 minutes followed by air line alkali wash. The alkali was recirculated.
- Alkali was drained using drain 2 valve and it was ensured that the steam traps were open so as to drain off any remnants.
7. Rinsing with purified water was done using recirculation
8. Acid wash (0.5% HNO3) was performed in the same manner as the alkali wash.
9. Rinsing with purified water was done as before.
10. W.F.I. rinse without recirculation.
- The first two rinses were for the transfer and air lines followed by the third rinse for the fermenter vessel.
- Contents drained using the drain 1 and drain 2 valves of the CIP skid.