Industrial Microbiology Dr. Butler 2011

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