Podcast
Questions and Answers
What is the primary purpose of using packed-bed or cartridge filters in air systems?
What is the primary purpose of using packed-bed or cartridge filters in air systems?
Which filter design is likely to improve the lifetime of a filter beyond three years?
Which filter design is likely to improve the lifetime of a filter beyond three years?
What is the recommended relative humidity for air provided by compressors?
What is the recommended relative humidity for air provided by compressors?
Which type of valve is commonly used for continuous sterilizing operations?
Which type of valve is commonly used for continuous sterilizing operations?
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For the efficient transfer of inoculum to a fermenter, which type of pump is commonly used?
For the efficient transfer of inoculum to a fermenter, which type of pump is commonly used?
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What is the bed depth range for packed-bed filters?
What is the bed depth range for packed-bed filters?
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Which material is commonly used for processing valves and piping?
Which material is commonly used for processing valves and piping?
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Why is it necessary to use heat exchangers for air coming from compressors?
Why is it necessary to use heat exchangers for air coming from compressors?
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What is the primary reason for cooling media in the fermentation process?
What is the primary reason for cooling media in the fermentation process?
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What can cause stress corrosion cracking in stainless steel during sterilization?
What can cause stress corrosion cracking in stainless steel during sterilization?
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What is the recommended dischargeable level of chloride content in cooling water?
What is the recommended dischargeable level of chloride content in cooling water?
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Which method is NOT suitable for managing odors from the fermentation process?
Which method is NOT suitable for managing odors from the fermentation process?
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How often should the chloride content in cooling water be checked?
How often should the chloride content in cooling water be checked?
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Which factor is a concern for the fermentation department regarding cooling water?
Which factor is a concern for the fermentation department regarding cooling water?
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What is a potential consequence of discharging cooling water into a stream without monitoring?
What is a potential consequence of discharging cooling water into a stream without monitoring?
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What is the purpose of using a conductivity probe in the cooling water line?
What is the purpose of using a conductivity probe in the cooling water line?
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What effect is produced when there is a change in the shape of a crystal?
What effect is produced when there is a change in the shape of a crystal?
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Which method is commonly used for controlling foam during microbial fermentation?
Which method is commonly used for controlling foam during microbial fermentation?
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What is one of the primary functions of safety valves in fermentation systems?
What is one of the primary functions of safety valves in fermentation systems?
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Which of the following is a source of contamination in the fermentation department?
Which of the following is a source of contamination in the fermentation department?
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Which device can be used to prevent foam build-up in fermenters?
Which device can be used to prevent foam build-up in fermenters?
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What issue can arise from the presence of foreign microorganisms in fermentation media?
What issue can arise from the presence of foreign microorganisms in fermentation media?
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What is a consequence of improper operator training in fermentation processes?
What is a consequence of improper operator training in fermentation processes?
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What is one primary factor that influences the concentration of dissolved oxygen during fermentation?
What is one primary factor that influences the concentration of dissolved oxygen during fermentation?
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What is the primary purpose of microbiological laboratories in bioprocessing?
What is the primary purpose of microbiological laboratories in bioprocessing?
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Which of the following is NOT a typical function of analytical support laboratories?
Which of the following is NOT a typical function of analytical support laboratories?
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Which equipment is commonly found in analytical support laboratories?
Which equipment is commonly found in analytical support laboratories?
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What component is essential for maintaining aseptic operations in fermentation?
What component is essential for maintaining aseptic operations in fermentation?
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Which area is likely to be found in a typical microbiological laboratory setup?
Which area is likely to be found in a typical microbiological laboratory setup?
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Which of the following is a method for measuring process variables in fermentation?
Which of the following is a method for measuring process variables in fermentation?
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What is a key role of bioinstrumentation in bioprocess design?
What is a key role of bioinstrumentation in bioprocess design?
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What is the purpose of using spargers in fermentation?
What is the purpose of using spargers in fermentation?
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What is the primary function of orifice spargers in fermentation processes?
What is the primary function of orifice spargers in fermentation processes?
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What is a significant advantage of the single-nozzle sparger over other sparger types?
What is a significant advantage of the single-nozzle sparger over other sparger types?
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What best describes the containment guidelines established during the 1970s?
What best describes the containment guidelines established during the 1970s?
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Which of the following criteria is NOT considered when classifying microorganisms into hazard groups?
Which of the following criteria is NOT considered when classifying microorganisms into hazard groups?
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In a combined sparger-agitator design, what is the purpose of the holes drilled in the disk between the blades?
In a combined sparger-agitator design, what is the purpose of the holes drilled in the disk between the blades?
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How are nongenetically engineered organisms classified into hazard groups?
How are nongenetically engineered organisms classified into hazard groups?
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What is a primary concern of aseptic operation in fermentation?
What is a primary concern of aseptic operation in fermentation?
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What does containment in fermentation processes primarily aim to prevent?
What does containment in fermentation processes primarily aim to prevent?
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Study Notes
Microbiological Laboratories
- Microbiological labs focus on strain improvement for increased yields, byproduct suppression, surfactant reduction, and broth modification for easier product recovery.
- Other ongoing research involves culture preservation techniques, stability testing, new procedure development, media optimization, and identification of inducers, repressors, and inhibitors.
- Lab space and equipment requirements include:
- Glassware and equipment washing area
- Media preparation area(s)
- Inoculation room(s)
- Incubator area(s)
- Office
- Laboratories
Analytical Support Laboratories
- Responsibilities include sterility testing, chemical assays for raw materials, materials before sterilization, optimized production batches, fermenter feeds, and waste streams.
- Common lab equipment includes auto-analyzers for product, carbohydrates, phosphate, various ions, specific enzymes, and other tools such as balances, gas chromatography (GC), high pressure liquid chromatography (HPLC), Kjeldahl equipment, titrimeters, UV-visible spectrophotometers, atomic absorption spectrophotometer (AAS), pH meters, viscometer, refractometer, densitometer, etc.
Air Filtration Systems
- Packed-bed or cartridge filters are used for sterile air and humidity reduction.
- Careful filter design extends the filter lifetime beyond 3 years.
- Fine fiber diameter in packed-bed filters increases filtration efficiency.
- Typical bed depth is 10 to 18 inches with 10 microns filter.
- Some plants utilize a separate filter for each sterile vessel, while others have a centralized filter system.
Air Compressors
- Oil-free compressed air is ideal, commonly provided by oil-free screw air compressors.
- Relative humidity should be about 85%.
- Heat exchangers are required to lower air temperature below the dew point and to reheat and control humidity.
Valves
- Gate, diaphragm, ball, and plug valves are common.
- While less of a concern for sterility, diaphragm and ball valves require maintenance.
- Diaphragm and ball valves are ideal for batch sterilizing operations.
- Plug-type valves are preferred for continuous sterilizing operations.
- Plug or diaphragm valves are common for inoculum transfer and sterile feed piping.
- All process valves and piping are 316 S/S.
Pumps
- Peristaltic pumps are a simple method for transferring inoculum between laboratory flasks, seed fermenters, and fermenters.
- Centrifugal pumps (316 S/S) are used to pump non-sterile raw materials, slurries, harvested broth, etc.
Cooling Equipment
- Cooling is required for media cooling, removal of fermentation heat, broth cooling before harvesting, and compressed air cooling.
- Cooling water is provided from cooling towers and chilled water (5º-15ºC) is produced by steam vacuum or refrigeration units.
- The fermentation department should be concerned about cooling water temperature and chloride content.
- Chloride ions above 150 ppm in water at temperatures above 80ºC can cause stress corrosion cracking of stainless steel.
- Conductivity probes are used for monitoring the cooling water line and prevent high salt levels.
- Discharge permits and monitoring may be required by pollution control agencies if cooling water is discharged to waterways.
- The chloride content in cooling water should be determined analytically every two weeks to maintain levels below 100 ppm.
Environmental Control
-
Odor:
- Sterilization of certain raw materials produces odors.
- High air discharge from a large fermenter house makes odor unavoidable.
- Wet scrubbing towers with sodium hypochlorite can be used for reduction but are expensive and produce waste.
- Tall exhaust stacks dilute the released gas before reaching ground but aren't acceptable by pollution control boards.
- Ozone treatment can be effective.
-
Organic Pollution:
- The fermentation department should monitor and control COD/BOD of liquid waste before discharge to the sewer.
Spargers
-
Ring Sparger:
- A perforated pipe positioned below the impeller in the form of crosses or rings.
- Size should be ¾ of impeller diameter.
- Air holes should be at least 6mm diameter.
-
Orifice Sparger:
- Used without agitation in yeast manufacture, effluent treatment, and single-cell protein production in air-lift fermenters.
-
Nozzle Sparger:
- Found in many agitated fermenters, from laboratory to production scale.
- Consists of a single open pipe or partially closed pipe for air bubble delivery.
- Low pressure loss and resistance to blockage.
-
Combined Sparger-Agitator:
- Developed by Herbert, Phipps, and Tempest (1965) for small-scale fermentation (1 dm3).
- Air is introduced through a hollow agitator shaft and released through holes in the disc between the blades.
- Provides efficient aeration in baffled vessels at varying agitator RPMs.
Aseptic Operation and Containment
- Aseptic Operation: Protection against contamination.
- Containment: Prevention of viable cell escape from fermenters or downstream equipment.
- Containment regulations were introduced in the 1970s.
- Organisms are classified into hazard groups based on potential risks.
Pressure Control
- Regulatory valves and pressure gauges are employed to maintain pressure in various components.
- Safety valves are incorporated in vessels and pipelines to release pressure upon exceeding the working pressure.
Foam Sensing and Control
- Uncontrolled foam formation can lead to oxygen deprivation.
- Antifoam agents are added when foaming reaches predetermined levels.
- Foam sensing and antifoam addition methods depend on the process and economic considerations.
- Mechanical antifoam devices include discs, propellers, brushes, hollow cones, rotating shafts, centrifugal separators, and jets spraying on deflectors.
Troubleshooting in Fermentation Plant
- Problems may stem from electrical, instrumentation, mechanical, physical, or microbial sources.
- Microbial contamination can result in:
- Unusable inoculum.
- Production inhibition in fermenters.
- Contaminated products.
- Fermentation broth that is difficult to filter or process.
Basic Sources of Contamination in Fermentation Departments
- Contamination of stock cultures due to poor techniques.
- Contamination from raw materials.
- Inadequate sterilization of equipment, air, or media.
- Insufficient procedures or inadequate operator training.
- Bacteriophages.
Six Categories for Regular Inspection to Prevent Contamination
- 1. Microbiological Laboratory: Ensure proper sterilization techniques, media preparation & control, and regular monitoring for contaminants.
- 2. Fermentation Department: Maintain proper air filtration, cleaning procedures, and environmental controls to minimize contamination risks.
- 3. Media Preparation: Strict adherence to sterilization protocols and monitoring for sterility are essential.
- 4. Sterilization: Regular maintenance of sterilization equipment, monitoring of sterilization cycles, and validating sterilization processes are critical.
- 5. Equipment: Strict cleaning and sterilization procedures for fermenters, pipes, and all processing equipment.
- 6. Personnel: Good hygiene practices, proper gowning procedures, and training in aseptic techniques are vital for avoiding contamination.
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