Biology Lab report ASAP 6 hours

Biol 390-Lab 3 Introduction to the Bioreactor

Maryville University

Objective Become familiar with a lab scale bioreactor

Background Bioreactors are vessels that support the growth of single-celled prokaryotic or eukaryotic cells. They can be used to increase the biomass of an organism, such as bakers yeast. In other cases the biomass of an organism is increased, then used to convert a pre-cursor to a product, such as in penicillin production. Fermentations can be either aerobic or anaerobic depending on the organism.

Vessels can be made of glass in small-scale bioreactor. In large scale reactors are typically made of stainless steel

Overview a. BioFlo/CelliGen 115 is a versatile fermentor/bioreactor that provides a fully equipped system in a compact package. It can be employed for batch, fed batch, or continuous culture with process control for pH, dissolved oxygen (DO), agitation, temperature, pump feed, antifoam and foam/level.

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Functions • Microbial metabolism and growth can be controlled though

o Agitation speed and type of agitator

o Temperature 20oC over coolant temperature and up to 70oC

o Aeration - Four gases can be introduced and their flow controlled:

§ Air

§ Nitrogen

§ Carbon dioxide

§ Oxygen

o pH can be controlled between 2 and 14 using a pH sensor. Control is maintained by a P&I (proportional and integral) controller, which operates the acid and base pumps.

o Dissolved oxygen can be controlled in the range of 0 to 200%. An electrode senses D.O and levels can be controlled by a P&I controller.

o Foam control – A foam/level sensor is present in the head-plate. The controller operates an antifoam pump. Pumps turn on and off.

o Exhaust system – gases and moisture are removed.

o Sampler – samples can be taken into a sample tube by closing the exhaust.

Headplate

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Touch Screen This has a summary screen for control of

o Agitation

o Temperature

o pH

o DO

o Air

o Summary screen

o Gauge screen

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Setpoint Screen

Before you start 1. Connect water to the system and turn it on. 2. Make sure the drain line is properly connected to the system.

1. Note we currently not using a water cooling system 3. Connect the quick-connect plastic water lines to the exhaust condenser. 4. Add glycerin to the thermowell and insert the temperature probe. 5. Make sure the motor is not connected. Turn the mains/power ON. 6. Set the TEMP setpoint to the desired working temperature. 7. Check that agitation (Agit) is in OFF mode. Connect the motor, then set

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agitation to the desired speed, and select Auto as its control mode. 8. Remove the shorting cap from the pH probe. Connect the pH cable to the pH

probe. 9. Remove the protective cap from the DO probe and connect the DO cable to the

DO probe. 10. If you have a water-jacketed vessel, be sure to refill the water jacket if required.

The DO polarographic probe will need to be connected for a minimum of six hours, to be properly polarized, before it can be correctly calibrated.

11. Calibrate the DO probe (see Section 7.3). 12. Set pH and DO to the desired setpoints 13. Set the pH control mode to Auto. 14. Set the DO control mode to Auto. 15. Open the PUMP screen and assign a pump to Acid and another pump to Base.

Turn the pumps ON. 16. If you are using oxygen, set the O2 control loop to the desired setpoint for

oxygen enrichment. If, however, you are using Air only, set the O2 setpoint to 0 (zero).

17. Set the O2 (or Air) control loop control mode to O2 Enrich. 18. Enable the pumps. 19. Go to the CASCADE screen and select the DO loop. 20. Set up cascades as desired.

Inoculation Using the septum port:

1. Aseptically remove the inoculum from its flask with the inoculation syringe. 2. Inject the inoculum through the septum in the inoculation port.

If you prefer to inoculate via an addition port, be sure to flame the connectors and use an inoculum flask as your “addition vessel”.

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BioFlo

®

/CelliGen

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115wM1369-0050 Operating manual

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