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Questions and Answers

What is the primary purpose of using packed-bed or cartridge filters in air systems?

  • To increase humidity in the air
  • To compress the air more efficiently
  • To enhance air temperature above the dew point
  • To obtain sterile air (correct)

Which filter design is likely to improve the lifetime of a filter beyond three years?

  • Careful selection of cartridge or packed-bed filter design (correct)
  • Simple cloth filters
  • Plastic disposable filters
  • Cartridge filters with fine fiber material

What is the recommended relative humidity for air provided by compressors?

  • 65%
  • 50%
  • 100%
  • 85% (correct)

Which type of valve is commonly used for continuous sterilizing operations?

<p>Plug valve (C)</p> Signup and view all the answers

For the efficient transfer of inoculum to a fermenter, which type of pump is commonly used?

<p>Peristaltic pump (D)</p> Signup and view all the answers

What is the bed depth range for packed-bed filters?

<p>10 to 18 inches (A)</p> Signup and view all the answers

Which material is commonly used for processing valves and piping?

<p>316 Stainless Steel (B)</p> Signup and view all the answers

Why is it necessary to use heat exchangers for air coming from compressors?

<p>To lower the air temperature below the dew point (C)</p> Signup and view all the answers

What is the primary reason for cooling media in the fermentation process?

<p>To cool media from sterilizing temperatures (D)</p> Signup and view all the answers

What can cause stress corrosion cracking in stainless steel during sterilization?

<p>High chloride levels combined with elevated temperatures (B)</p> Signup and view all the answers

What is the recommended dischargeable level of chloride content in cooling water?

<p>Less than 100 ppm (C)</p> Signup and view all the answers

Which method is NOT suitable for managing odors from the fermentation process?

<p>Direct release of gases into the atmosphere (D)</p> Signup and view all the answers

How often should the chloride content in cooling water be checked?

<p>Every two weeks (B)</p> Signup and view all the answers

Which factor is a concern for the fermentation department regarding cooling water?

<p>Chloride content and temperature (B)</p> Signup and view all the answers

What is a potential consequence of discharging cooling water into a stream without monitoring?

<p>Violation of pollution control regulations (A)</p> Signup and view all the answers

What is the purpose of using a conductivity probe in the cooling water line?

<p>To indicate the salt level in the water (B)</p> Signup and view all the answers

What effect is produced when there is a change in the shape of a crystal?

<p>Piezoelectric effect (B)</p> Signup and view all the answers

Which method is commonly used for controlling foam during microbial fermentation?

<p>Adding antifoam agents (A)</p> Signup and view all the answers

What is one of the primary functions of safety valves in fermentation systems?

<p>To release excess pressure (D)</p> Signup and view all the answers

Which of the following is a source of contamination in the fermentation department?

<p>Poor sterilization techniques (B)</p> Signup and view all the answers

Which device can be used to prevent foam build-up in fermenters?

<p>Discs and propellers (C)</p> Signup and view all the answers

What issue can arise from the presence of foreign microorganisms in fermentation media?

<p>Contamination of final products (D)</p> Signup and view all the answers

What is a consequence of improper operator training in fermentation processes?

<p>Increased contamination risks (D)</p> Signup and view all the answers

What is one primary factor that influences the concentration of dissolved oxygen during fermentation?

<p>Foam formation (C)</p> Signup and view all the answers

What is the primary purpose of microbiological laboratories in bioprocessing?

<p>To perform mutation and isolation work for better strain yields. (A)</p> Signup and view all the answers

Which of the following is NOT a typical function of analytical support laboratories?

<p>Development of new media preparation techniques. (A)</p> Signup and view all the answers

Which equipment is commonly found in analytical support laboratories?

<p>High pressure liquid chromatography (HPLC). (C)</p> Signup and view all the answers

What component is essential for maintaining aseptic operations in fermentation?

<p>Proper containment protocols to prevent contamination. (C)</p> Signup and view all the answers

Which area is likely to be found in a typical microbiological laboratory setup?

<p>Media preparation area. (B)</p> Signup and view all the answers

Which of the following is a method for measuring process variables in fermentation?

<p>Online pH monitoring. (B)</p> Signup and view all the answers

What is a key role of bioinstrumentation in bioprocess design?

<p>Automating data collection and process control. (D)</p> Signup and view all the answers

What is the purpose of using spargers in fermentation?

<p>To distribute gases evenly throughout the fermenter. (A)</p> Signup and view all the answers

What is the primary function of orifice spargers in fermentation processes?

<p>Providing a point-source stream of air bubbles (A)</p> Signup and view all the answers

What is a significant advantage of the single-nozzle sparger over other sparger types?

<p>Lower pressure loss (D)</p> Signup and view all the answers

What best describes the containment guidelines established during the 1970s?

<p>They assess hazards to determine the necessary degree of containment. (C)</p> Signup and view all the answers

Which of the following criteria is NOT considered when classifying microorganisms into hazard groups?

<p>Availability of vaccines (C)</p> Signup and view all the answers

In a combined sparger-agitator design, what is the purpose of the holes drilled in the disk between the blades?

<p>To provide aeration by emitting air (C)</p> Signup and view all the answers

How are nongenetically engineered organisms classified into hazard groups?

<p>Using criteria to assess their risk (C)</p> Signup and view all the answers

What is a primary concern of aseptic operation in fermentation?

<p>Protection against contamination (C)</p> Signup and view all the answers

What does containment in fermentation processes primarily aim to prevent?

<p>Escape of viable cells from the fermenter (D)</p> Signup and view all the answers

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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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