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CEN 551 Biochemical Engineering

CEN 551 Biochemical Engineering. Instructor: Dr. Christine Kelly. Class Outline. Syllabus and course format introductions Penicillin Types of microorganisms Central dogma. Syllabus. Bioprocess Engineering: Basic Concepts, second edition. M. L. Shuler and F. Kargi. Prentice Hall.

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CEN 551 Biochemical Engineering

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  1. CEN 551 Biochemical Engineering Instructor: Dr. Christine Kelly

  2. Class Outline Syllabus and course format introductions Penicillin Types of microorganisms Central dogma

  3. Syllabus • Bioprocess Engineering: Basic Concepts, second edition. M. L. Shuler and F. Kargi. Prentice Hall. • Grading 3 exams: 12% each, total 36% of course grade Assignments: total of 30% of course grade Project: 34% of course grade

  4. Course Format • PowerPoint-based lectures. I will provide handouts of the lectures and put the lectures on the web. • We will cover the introductory chapters (1-7) quickly, and focus on the engineering portion of the text (chapters 8, 9, 10, 11, 12, 14 and 15). We will not cover chapters 13 and 15.

  5. Introductions Are you a graduate student/undergraduate student? Why are you taking this course? What is your background in biology?

  6. Relationship of Scientists Engineers • Microbiologists, biochemists, and molecular biologists are scientists, well-trained in empirical testing of hypotheses. • Engineers develop theories based on mathematical models, use models to predict performance, optimize and develop processes.

  7. Biologists and Engineers • Research scientists often pursue knowledge while applications may take a secondary role. • The work of engineers is often driven by economics of an application and problem solving.

  8. Penicillin: “Birth of Biochemical Engineering” • 1928- Alexander Flemming was plating Staphylococcus aureus and the plate was contaminated with mold – near the mold no bacteria grew. • WWII- most common cause of death was infection from wounds. • Sulfa drugs were effective on limited range of infectious organisms. • 1930-1940 British scientists Florey and Chain at Oxford developed a process to produce penicillin from the mold.

  9. How Penicillin Works... Antibiotics on a plate: cell walls do not form

  10. Early Work • They asked US pharmaceutical companies to help work on the project – to develop a commercial scale process for penicillium. • Merck, Pfizer, Squibb, USDA • At this time, most drugs were made synthetically. Fermentation was unproved and companies were skeptical. • Problem: low concentrations, fragile product.

  11. Significant Advances • New medium- Corn steep liquor (x10). • New strain isolated from molded fruit- P. chrysogenum (still used in some form). • Change to tanks from “bottle plants”. • Separation: liquid-liquid extraction.

  12. Challenges • Very large (10 kgal) fermentation vessels. • Provide sterile air and feed. • Agitator seal. • Heat removal. • Recovery and purification of fragile product.

  13. Biology-Engineering Connection • Cooperation between engineers and scientists was critical (Merck specifically formed teams of each). • “Biochemical engineering” born as a result.

  14. Related WWW Resources • What the heck is…” page: http://people.ku.edu/~jbrown/whatheck.html • Penicillin: http://people.ku.edu/~jbrown/penicillin.html

  15. Source for Figures Most of the figures in this lecture come from an excellent web site that you should examine. http://www.bact.wisc.edu/microtextbook/index.html

  16. Naming Conventions • Microorganisms are commonly named using genus and species (ex. Escherichia coli or E. coli; note italics, capital on genus, lower case on species). • After species, arbitrary designations (ex. E. coli HCB457)

  17. Extremophiles • Thermophiles, acidophiles, halophiles, psychrophiles • E. coli dies at 66˚C, thermophiles can live up to 110˚C.

  18. Principal Cell Types • Procaryotes • genetic material not in membrane • one circular chromosome, no organelles • Eucaryotes • nuclear membrane • >1 chromosome

  19. Viruses- D/RNA looking for a home • replicate by pirating host cells’ protein manufacturing mechanisms • bacteriophage (phage) infect bacteria • bacteriophage very useful for genetic engineering as genetic “vector” • host cells lyse or reproduce with viral DNA

  20. Procaryotes • Usually from 0.5 to 3 µm, can be >600µm • Eubacteria (true bacteria) • spherical, rod-like, spiral-shaped • chemistry like eucaryotes • often divided into gram + or gram -

  21. Characteristics of Procaryotes

  22. Common Procaryote Cell ShapeVariations

  23. Gram Positive Eubacteria

  24. Gram Negative Eubacteria

  25. Intracellular Material in Procaryotes • ribosomes • site of protein synthesis • storage granules • PHB, other polysacc. storage sites

  26. Bacterial Spores (endospores) • formed inside cells of some strains • defense against unfavorable growth conditions • can survive hours of boiling • generate vegetative (normal) cells when conditions allow

  27. Extracellular Material • exopolysaccharides • principle component of biofilms • fimbria (some are “sex organs”) • flagella

  28. Bacterial Flagella

  29. Archaebacteria (Archae) • no peptidoglycan (component of cell wall in eubacteria) • composition of cytoplasmic membrane different • as a result, are extremophiles

  30. Eucaryotes: Fungi, algae, protozoa • Plant and animal cells are also eucaryotic cells • on average, larger than procaryotes • have nuclear membrane, organelles • DNA in linear strands • replication by sexual & asexual means • may also have flagella or cilia

  31. Plant and Animal Cells • Plant cells contain chloroplasts, where photosynthesis occurs • Animal cells have no cell walls and are very sensitive to shear.

  32. Eucaryotic Cells

  33. Eucaryotes: Fungi • Yeasts • Round or oval, and typically 5-10 microns in size • reproduce by budding, fission (both asexual) or fusion (sexual reproduction) • S. cerevisiae- used to produce ethanol and in cooking.

  34. Eucaryotes: Fungi • Molds • multiple nuclei in cytoplasm • filamentous, can be grown in stirred liquid medium • asexual and sexual reproduction by spores • production of citric acid and penicillin

  35. Eucaryotes: Algae and Protozoa • Protozoa are important higher organisms in waste treatment • Algae are major contributors to fouling if light is available • Currently, lesser players in the bioprocess industry • used in agar, filtration industry (silica-containing algae)

  36. Central Dogma(might as well memorize now!) • Replication • Transcription • Translation

  37. More Review

  38. Cell Construction Living cells: bioreactors with >2000 (mostly coupled) reactions Proteins Lipids Carbohydrates Nucleic acids

  39. Proteins

  40. Amino Acids and Proteins • Proteins account for 40%-70% of a cell’s dry weight. • Proteins consist of chains of amino acid subunits.

  41. Amino Acid a carbon

  42. Formation of a Protein Peptide bonds between amino acids

  43. Charge and Amino Acids • Acidic and basic groups lead to different charges at different pH values. • Overall charge can be negative (anionic), positive (cationic), or have an overall charge of zero (zwitterionic form). • pH dependence of charge often exploited in separation of amino acids

  44. Functional Classes of Proteins • Catalytic- enzymes (highly specific) • Structural- collagen • Transport- hemoglobin • Regulatory- hormones • Protective- antibodies

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