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Bioreactor: Optimizing E.Coli fermentation for Bacteria directed enzyme prodrug therapy

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  1. Introduction
  2. Design of the study
    1. Title of the study
    2. Objectives of the study
    3. Scope of the study
  3. Methodology of the study
    1. Research design of the study
  4. Sources of data
    1. Primary source
    2. Secondary source
  5. Limitations of the study
  6. Theoritical background of the study
    1. Causes of stress in the workplace
    2. Job stress and health
  7. Work/life conflicts
  8. Eliminating barriers to advancement
    1. Preparing women for management
    2. Accommodating families
    3. Work-life practice guidelines
  9. Brief of companies policies and facilities
    1. Infosys
    2. Wipro
    3. SAP
    4. IBM
    5. Google
  10. Findings of the study
  11. Recommendations
  12. Bibliography

E.coli strain WM04 was determined as a viable option for Bacteria Directed Enzyme Prodrug Therapy. Determination of suitability for animal testing depends on sufficient quantities of bacteria being produced through fermentation. Optimal operating conditions are found for the production of WM04 by observing two important production variables: the quantity of bacteria produced and the capacity for survival of the bacteria.

One limitation to bacterial growth is the amount of oxygen supply. The concentration of dissolved oxygen and oxygen transfer rates are correlated to cell growth. Impellor velocity, ranging from 200-1000 rpm, allowed us to determine the optimal speed at which the greatest oxygen transfer rate was achieved in the bioreactor solution with minimal bacterial shearing.

The dissolved oxygen content of the media in the bioreactor was measured with an oxygen probe at varying impellor speeds. BioCommand Lite software was used to plot the dissolved oxygen concentration against the time of the run and the impellor speed versus time.

[...] Introduction The E.coli strain WM04 has been bioengineered for Bacterial Directed Enzyme Prodrug Therapy for laboratory testing. Preliminary testes for WM04 production were conducted in shake flasks; however we want to use a bioreactor to carry out larger-scale growth. WM04 was altered to produce an antibody on its surface, which recognizes a tumor antigen. This membrane modification increases the sensitivity of the bacterial membrane to shearing at high impellor speeds. We want to determine the optimal operating conditions to minimize negative effects of shearing while still producing sufficient amounts of bacteria for use in animal testing. [...]

[...] A number of standard curves for determining cell concentration were generated by measuring the optical density and counting plated colonies of dilute bacterial concentrations. An initial concentration of 1x108 cells/mL was assumed for the overnight E. coli culture stock5. Figure 3 shows the relationship between the dilution factor of the stock culture and the optical density of the corresponding samples. Figure 3. Relationship between dilution factor of samples and optical density. With very dilute solutions, the absorbance plateaus as the dilution factor increases. [...]

[...] Thus, the media provided a steady nutrient supply for E.coli growth for this fermentation process. The calculations and data obtained experimentally showed that running the bioreactor at 200 rpm generated the maximum amount of bacterial cells. Despite the lower oxygen transfer rate at 200 rpm, it did not significantly affect the generation of the bacteria. The shear from the higher impellor speed and oxygen transfer rate caused more damage to the bacterial cells, resulting in a decrease in cell generation. [...]

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