Lecture 1 introduction
- 1. Industrial Microbiology MBIO 4510
- 2. Lecture 1 – Introduction to Industrial Microbiology What is Industrial Microbiology? Industrial microbiology is the commercial exploitation of microorganisms to produce valuable economic, environmental and socially important products, or to carry out important chemical transformations.
- 3. Lecture 1 – Introduction to Industrial Microbiology Madigan, M.T. 2003. Brock Biology of Microorganisms 10th ed. New Jersey: Prentice Hall. P 967
- 4. Lecture 1 – Introduction to Industrial Microbiology Fermentation Products Food, beverage, food additives and supplements Dairy products (yogurt, cheese) Alcoholic beverages (beer, wine) Amino acids, vitamins
- 5. Lecture 1 – Introduction to Industrial Microbiology Fermentation Products Health Care Products Antibiotics – over 4000 isolated, only 50 used regularly • β-lactams, penicillins, and cephalosporins • Aminoglycosides (streptomysin) • Tetracyclins Important to develop new antibiotics due to abuse/misuse of current antibiotics
- 6. Lecture 1 – Introduction to Industrial Microbiology Fermentation Products Health Care Products Alkaloids, steroids, vaccines Therapeutic recombinant human proteins (insulin, interferons, blood-clotting factors, human growth hormone) More recombinant therapeutic products to be developed
- 7. Lecture 1 – Introduction to Industrial Microbiology Fermentation Products Production of microbial enzymes Proteases, carbohydrases, Taq polymerase Industrial chemicals and fuel Methane, ethanol, H2, propane, etc. Environmental roles of microorganisms Waste water treatment, desulphurization of fuels, leaching of metals, use of microbes to reduce usage of synthetic pesticides
- 8. Lecture 1 – Introduction to Industrial Microbiology Overview of a Fermentation Process Waites et al. 2001. Industrial Microbiology: An Introduction. Oxford: Blackwell Science. P 2
- 9. Lecture 1 – Introduction to Industrial Microbiology Fermentation process – Upstream Processing 1. Fermentation Organism need suitable cells to produce desired products (bacteria, fungi, yeast, animal cells) improve strain to enhance productivity and yield maintain purity of cultures
- 10. Lecture 1 – Introduction to Industrial Microbiology Fermentation process – Upstream Processing Waites et al. 2001. Industrial Microbiology: An Introduction. Oxford: Blackwell Science. P 83
- 11. Lecture 1 – Introduction to Industrial Microbiology To be useful for commercial processes, cells must: produce usable products or effects be available in pure culture be genetically stable, or genetically mutated produce spores or other reproductive structures to allow easy inoculation grow rapidly and produce product quickly in large scale culture* be easily separated from products not be harmful to humans, plants, animals, etc
- 12. Lecture 1 – Introduction to Industrial Microbiology Fermentation process – Upstream Processing 2. Fermentation Medium need cost-effective carbon and energy sources, essential nutrients media often wastes from other processes, such as sugar processing wastes, lignocellulosic wastes, cheese whey and corn steep liquor
- 13. Lecture 1 – Introduction to Industrial Microbiology Fermentation process – Upstream Processing 3. Fermentation industrial microorganisms cultivated under controlled conditions to optimize growth of organism and production of microbial products must avoid environmental conditions that trigger regulatory mechanisms (repression, feedback inhibition)
- 14. Lecture 1 – Introduction to Industrial Microbiology Fermentation process – Upstream Processing Madigan, M.T. 2003. Brock Biology of Microorganisms 10th ed. New Jersey: Prentice Hall. P 970
- 15. Fig. 9.2
- 16. Fig. 9.3
- 17. Fig. 9.4
- 18. Genentech 12,000 L animal cell bioreactor
- 19. Lecture 1 – Introduction to Industrial Microbiology Fermentation process – Downstream Processing includes all processes after fermentation involve cell harvesting, cell disruption, product purification from cell extracts or the growth medium must be rapid and efficient to purify product and to maintain stability of product safe and inexpensive to dispose of wastes
- 20. Lecture 1 – Introduction to Industrial Microbiology Fermentation Products Primary Metabolites: produced during active growth (trophophase) amino acids, organic acids, alcohol fermentation products, vitamins Secondary Metabolites: produced during stationary phase after microbial biomass production has peaked (idiophase) generally not essential for growth or reproduction antibiotics, citric acid
- 21. Lecture 1 – Introduction to Industrial Microbiology Fermentation process Waites et al. 2001. Industrial Microbiology: An Introduction. Oxford: Blackwell Science. P 24
- 22. Lecture 1 – Introduction to Industrial Microbiology Fermentation Products Madigan, M.T. 2003. Brock Biology of Microorganisms 10th ed. New Jersey: Prentice Hall. P 968
- 23. Lecture 1 – Introduction to Industrial Microbiology Fermentation Products Economics of fermentation determined by cost of raw materials, utilities, labour and maintenance, fixed charges, working capital charges, etc. High volume, low value products Products Low volume, high value products
- 24. Scale up? $$$$ 60 million of patients 12 Clinical Trials 500 Undisclosed 21 Yeast 50 30 Mammalian 25 39 20 15 Prokaryotic US Billion 10 5 0 2001 2004
- 25. Therapeutic Monoclonal Antibodies Datamonitor report “Mabs are hottest segment of biotech industry” articles in “Fierce Biotech” and “Bioprocess International” Mabs generate revenue of $20 billion 14% annual growth expected 2006-2012 and outstrips other sectors of pharmaceutical industry (Avastin, Herceptin, Remicade, Rituxan, Humira , and Erbitux) are 6 blockbusters.
- 27. The demand for mammalian cell culture products 60 3000 50 number of products 2500 Cumulative product approvals Kg capacity demand 40 2000 Demand (kg) 30 1500 20 1000 10 500 0 0 1986 1988 1990 1992 1994 1996 1998 2000 2002 2004 2006 2008 Butler, M. (2005) Applied Microbiology and Biotechnology 68: 283-291.
- 28. Pharmaceutical Prices 1e+9 1e+8 1e+7 epo 1e+6 infergen 1e+5 abciximab basiliximab herceptin 1e+4 enbrel Price ($/g) rituximab 1e+3 humulin 1e+2 1e+1 Plasma HSA 1e+0 1e-1 penicillin 1e-2 lysine 1e-3 ethanol 1e-4 1e-3 1e-2 1e-1 1e+0 1e+1 1e+2 1e+3 1e+4 1e+5 1e+6 1e+7 1e+8 1e+9 Annual demand (kg)
- 29. Lecture 9 Animal Cell Biotechnology Scaling up the production process Butler, M. 2004. Animal cell culture and technology 2nd ed. London and New York:Garland Science/BIOS Scientific Publishers. P203.
- 30. Pre-purification vs selling price of biological products Concentration in starting medium (g/l) 1e+3 ethanol 1e+2 citric acid amino acids 1e+1 penicillin 1e+0 bulk enzymes 1e-1 insulin 1e-2 m.antibodies 1e-3 1e-4 1e-5 factor VIII 1e-6 therapeutic enzymes 1e-7 1e-2 1e-1 1e+0 1e+1 1e+2 1e+3 1e+4 1e+5 1e+6 1e+7 1e+8 1e+9 1e+10 Selling price ($ per kg)
- 31. Wurm,F (2004) Nature Biotech 22: 1393
- 32. Milestones in the development of animal cell technology 1880 Roux maintained embryonic chick cells in saline solution 1890 1900 Harrison grew frog nerve cells by the 'hanging drop' technique. 1910 Carrel used aseptic techniques for long term cell cultures. Rous and Jones used trypsin for sub-culture of adherent cells. 1920 The 'Carrel' flask was designed for cell culture. 1930 1940 Antibiotics were added to culture medium. Earle isolated mouse L fibroblasts. Enders grew polio virus on cultured human cells. 1950 Gey cultured HeLa cells.
- 33. 1960 Hayflick and Moorhead showed that human cells have a finite lifespan. Ham grew cells in a serum-free medium. Harris and Watkins fused human and mice cells. 1970 Kohler and Milstein produced an antibody-secreting hybridoma. Sato developed serum-free media from hormones and growth factors. 1980 Human insulin was produced from bacteria. Monoclonal antibody (OKT3) used for human therapy. Recombinant tPA licensed for human therapy. 1990 Humanized chimeric antibodies used for human therapy Stem cells isolated