CBE661 Bioprocess Engineering Lab UITM Assignment Sample Malaysia

CBE661 Bioprocess Engineering Lab is a course that is typically offered at the undergraduate or graduate level in chemical and biochemical engineering programs. The course focuses on providing students with hands-on experience in designing, operating, and optimizing bioprocesses.

The course may involve laboratory experiments where students work with microbial cultures and perform various operations such as sterilization, media preparation, and bioreactor operation. Students may also analyze data generated from experiments and use software tools to simulate and optimize bioprocesses.

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In this section, we discuss some assignment briefs. These are:

Assignment Brief 1: Operate shake flask and bioreactor fermentation, prepare culture media, construct a growth curve and calculate mass transfer coefficient in fermentation.

Operating Shake Flask and Bioreactor Fermentation:

Shake Flask Fermentation:

  • Prepare the culture media according to the requirements of the organism to be cultured.
  • Sterilize the media and distribute it into shake flasks.
  • Inoculate the shake flasks with the organism to be cultured.
  • Incubate the shake flasks in a shaking incubator at the appropriate temperature, pH, and aeration conditions for the organism.
  • Monitor the growth of the organism by taking samples at regular intervals and measuring the optical density or other growth parameters.
  • Harvest the cells at the desired time point for downstream applications.

Bioreactor Fermentation:

  • Prepare the culture media according to the requirements of the organism to be cultured.
  • Sterilize the media and the bioreactor vessel.
  • Inoculate the bioreactor with the organism to be cultured.
  • Set the operating conditions for the bioreactor, such as temperature, pH, agitation speed, and aeration rate.
  • Monitor the growth of the organism by taking samples at regular intervals and measuring the optical density or other growth parameters.
  • Control the operating conditions to maintain optimal growth conditions for the organism.
  • Harvest the cells at the desired time point for downstream applications.

Preparing Culture Media:

  • Determine the nutrient requirements of the organism to be cultured.
  • Select appropriate ingredients and prepare the media according to established protocols.
  • Sterilize the media by autoclaving or filtration.
  • Adjust the pH and other parameters as necessary.

Constructing a Growth Curve:

  • Inoculate the organism in a suitable culture vessel (shake flask or bioreactor) with the appropriate media.
  • Measure the optical density or other growth parameters (e.g., dry weight, cell count, protein content) at regular intervals.
  • Plot the growth data over time to construct a growth curve.
  • Analyze the growth curve to determine the growth rate and other growth parameters.

Calculating Mass Transfer Coefficient in Fermentation:

  • The mass transfer coefficient is a measure of how efficiently oxygen is transferred from the air to the culture medium.
  • It can be calculated by measuring the oxygen transfer rate (OTR) and the volumetric oxygen transfer coefficient (kLa).
  • The OTR can be measured using an oxygen probe and the kLa can be determined experimentally by varying the agitation rate and/or aeration rate.
  • The mass transfer coefficient (kLa) can then be calculated using the following equation:
    kLa = OTR / (P_O2,sat – P_O2)

    1. OTR is the oxygen transfer rate
    2. P_O2,sat is the saturation concentration of oxygen in the medium
    3. P_O2 is the partial pressure of oxygen in the medium.

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Assignment Brief 2: Extract soluble enzyme from fungi or bacteria, utilize the enzyme in soluble or immobilized form and calculate the enzyme activity and kinetics.

Extracting Soluble Enzyme from Fungi or Bacteria:

  1. Choose a suitable microorganism that produces the desired enzyme.
  2. Grow the microorganism in a suitable medium that supports its growth and enzyme production.
  3. Harvest the microorganism at the appropriate stage of growth.
  4. Disrupt the cells to release the enzyme. This can be achieved by various methods such as sonication, homogenization, or enzymatic digestion.
  5. Centrifuge the lysate to remove cellular debris and collect the supernatant containing the soluble enzyme.
  6. Store the enzyme at an appropriate temperature and pH until further use.

Utilizing the Enzyme in Soluble or Immobilized Form:

  1. Determine the optimal pH and temperature for enzyme activity.
  2. Choose a suitable substrate and measure its concentration.
  3. Add the enzyme to the substrate solution and incubate for a specific period.
  4. Measure the product concentration at regular intervals using a spectrophotometer or other appropriate method.
  5. Repeat the experiment with varying enzyme concentrations to determine enzyme activity.
  6. Immobilize the enzyme on a suitable support material such as agarose beads, silica gel, or magnetic nanoparticles, using a suitable method such as covalent bonding, adsorption, or encapsulation.
  7. Measure the enzyme activity of the immobilized enzyme using the same procedure as for the soluble enzyme.

Calculating the Enzyme Activity and Kinetics:

  1. Calculate the initial reaction rate using the slope of the linear part of the product concentration vs. the time curve.
  2. Determine the Michaelis-Menten constant (Km) and maximum reaction rate (Vmax) using Lineweaver-Burk or other appropriate kinetic analysis methods.
  3. Determine the effect of pH and temperature on enzyme activity and calculate the activation energy using the Arrhenius equation.
  4. Determine the effect of substrate concentration on enzyme activity and calculate the Hill coefficient to determine the degree of cooperativity.
  5. Compare the properties of the soluble and immobilized enzymes and determine the advantages and disadvantages of each form.

Assignment Brief 3: Differentiate the mechanism involved in cross flow filtration and in freeze and spray drying processes and predict the possibility factors the affecting the processes.

Cross-flow filtration and freeze/spray drying are two different processes used in the separation and drying of various products. The mechanisms involved in these processes are different, as are the factors that affect their efficiency and outcome.

Cross-flow filtration is a process that uses a membrane to separate a mixture into its individual components based on size and molecular weight. In this process, the mixture is fed into the filtration system, where it is pumped through the membrane. The membrane is designed to allow the smaller particles to pass through while retaining the larger particles. The filtrate is collected on one side of the membrane, and the retained particles are collected on the other side. This process is used for various applications such as water purification, protein concentration, and blood plasma separation.

On the other hand, freeze and spray drying are two different methods used to remove moisture from a product. In freeze drying, the product is first frozen, and then the moisture is removed by sublimation under a vacuum. Spray drying, on the other hand, involves the spraying of a liquid mixture into a hot chamber where the liquid droplets are converted into dry particles by evaporation. Both methods are commonly used in the food, pharmaceutical, and chemical industries.

The factors that affect the efficiency and outcome of these processes are different. In cross-flow filtration, the membrane’s quality, size, and pore size can affect the separation efficiency. Other factors such as feed concentration, flow rate, and temperature can also impact the process. In freeze and spray drying, the product’s composition, particle size, and viscosity can affect the drying efficiency. Other factors such as drying temperature, air velocity, and residence time can also affect the process.

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