1. Biochemical Engineering

2. Outline What is biochemical engineering? The catalysts Relationship to engineering Historical perspective: penicillin The industry: facts The industry: Food, beer and wine Industrial chemicals Pharmaceuticals

3. Some definitions…. Biotechnology: use or development of methods of direct genetic manipulation for a socially desirable goal. Sometimes a broader definition is used, where biotechnology is applied biology. Biomedical Engineering: engineering on systems to improve human health Bioengineering, biological engineering: work on medical or agricultural systems, draws on electrical, mechanical, industrial and chemical engineers. Biochemical Engineering: extension of chemical engineering principles to systems using a biocatalyst to bring about desired chemical transformation.

4. The story of penicillin Alexander Fleming's photo of the dish with bacteria and Penicillin mold, 1928

5. The story of penicillin 19281940’s Discovered by Alexander Fleming Chemical synthesis proved to be too difficult Fermentation route was chosen Efforts to increase production cell line selection medium optimization process development Full scale production 15+ years

6. Biocatalyst: cells and enzymes 10-20  m 1-5  m

7. Enzyme: Acetylcholinesterase size range in nm E + S E-SE + P

8. Typical chemical processing A B A+B P PABPAB temperature flowrate A P B ReactorSeparation

9. Cell as a bioreactor Product : cells small molecule enzyme A P C D etc. ? Reactor inside a reactor Separation reactor A P cell B C D E P A

10. Bioprocesses are “special”……. Cells are living organisms that require specific conditions for growth, production Cells grow in culture medium (nutrients) that may support other cell types (contamination) Bioproducts are often sensitive to external conditions (T, pH) and can easily be destroyed (separations) Product is generally very dilute in bioreactor medium Many byproducts in medium to remove

11. ….but they have their advantages! Cells will often perform reactions that are too difficult to do synthetically (penicillin) Cells can turn basic nutrients (for example, agricultural waste) into valuable products Amazing diversity of products from cells Cells can be modified to increase product diversity even more! Enzymes are highly specific catalysts with high catalytic power

12. Industry B.Sc.M.S.Ph.D. 200019912000199120001991 Chemical 26.743.724.138.128.446 Fuels 12.621.28.516.17.615.3 Electronics 15.62.422.83.2194.8 Food/Consumer Products 11.47.24.5 2.85.6 Materials 3.32.77.15.86.25.6 Biotech and Related Industries 6.93.114.77.719.44 Pulp and Paper 2.45.11.33.20.90.8 Engineering Services 9.88.210.714.96.611.3 Business Services and Other* 11.26.46.36.496.5 AIChE Career Services Dept, 8/25/00 Engineering Employment Trends –

13. Industry Focus: Food and beverage

14. Fermentation Products cheese soy products yoghourt wine, beer bread Enzymes adjust food flavour adjust food texture improve nutritional quality high fructose corn syrup

15. Fermentation A form of anaerobic respiration occurring in certain microorganisms (ex. yeasts) Alcoholic fermentation is a series of biochemical reactions by which pyruvate is converted to ethanol and CO 2.

16. Metabolic pathways in e.coli

17. C 6 H 12 O 6 → 2 C 2 H 5 OH + 2 CO 2 Common yeast saccharomyces cerevisae used in making wine, beer, bread by above reaction. Different strains of yeast can tolerate different alcohol concentrations. Theoretically, 180 g of sugar will produce 92 g of ethanol Actual yield is only 84.6 g of ethanol  EtOH (20  C) = 0.789 g/mL volume of ethanol = 84.6 g x mL = 107.2 mL 0.789 g volume of the alcohol and water contracts by 0.7% so 107.2 mL x 1.007 = 108 mL for an overall alcohol concentration of 10.8% (v/v:108/1000).

19. Brix (B 0 ) is a density measurement that indicates the percentage of sugar in 100 g of a sugar-water solution. Brix can be calculated by: Brix = [g sugar/(g sugar + g water)] x 100 Brix units can be used to predict the alcohol content in wine. % potential alcohol (v/v) = 0.57 x Brix initial

20. Beer making Malting: grains (barley, rice) are steeped in water until germination and then dried before a plant develops. The starches in the grains get converted to sugars by enzymes. Brewing: finely ground malt is turned into a sweetened liquid by adding warm water added and heating to around 75o where the sugars get dissolved. Grain is filtered out, and its boiled for sterilization and concentration (wort) Fermentation: the yeast turns the sugar in the wort into alcohol, a process that takes about 10 days.

21. Wine making (1) Yeast Proliferation - aerobic oxygen is needed to sterol production increased yeast robustness (2) Initial Fermentation - anaerobic sugar is converted to alcohol duration times typically 5 - 12 days (3) Secondary Fermentation - malolactic lactic bacteria metabolize malic acid to lactic acid lowered acidity and wine “softening” occurs very typical for red wines

22. Process flowsheet for wine

23. Factors affecting yeast fermentation Yeast Species Temperature Sugar concentration pH Vitamins Sulfur dioxide (SO 2 )

24. Industry Focus: Textiles

25. Stone washing denim Denim is faded by abrasive action of pumice stones Indigo dye adheres to denim surface Cellulase enzyme removes some of the dye by partially hydrolyzing the cotton surface new looks lower costs shorter treatment times less solid waste weakens the fabric traditional method new method

26. Detergents Detergent industry is the largest single market for enzymes at 25 - 30% of total sales Dirt comes in many forms and includes proteins, starches and lipids (fats and oils) proteases, amylases, lipases are enzymes used in detergents enzymes allows lower temperatures and less agitation for washing Inner core of enzyme plus preservative bound with CMC Protective waxy coat that disperses in the wash

27. Industrial Chemicals Examples: organic acids produced from Aspergillus niger, citric acid used in soft drinks Xylanase used for wood pulping and bleaching

28. Agricultural Examples: Recombinant bovine somatotropin (bST) for increasing milk production Bio-insecticides for crop protection Phyto-vanilla(tm) flavor derived from tissue culture

29. Environment Cleanup of hazardous waste sites using bacteria that feed on pollutants Bacteria used for bio-remediation wastewater treatment Biosensors: use biological activity to detect toxic substances RIS® Water tests: antibody based kit to detect low level of solvents such as benzene

30. Fuel ethanol

31. Industry Focus: Pharmaceuticals

32. Phase II clinical trials in 100 to 300 patients The Drug Development and Approval Process The drug discovery and approval process takes and average of 15 years and costs almost $400 million source: Pharmaceutical Research and Manufacturers of America, Washington DC, 1996 Discovery of a promising compound Preclinical testing in animals Phase III clinical trials in 1000 to 3000 patients FDA review and approval Drug may be prescribed by physicians 1 year 3 years19 months 6.5 years2 years Phase I clinical trials in healthy volunteers

33. Products Small molecules and metabolites antibiotics Protein drugs Vaccines Antibodies, Monoclonal antibodies (MAb) How? Recombinant DNA technology means bacteria and yeast can produce human proteins like insulin

34. Penicillin fermentation

35. 2005 World market for biopharmaceuticals Enbrel Rheumatoid arthritis and psoriasis Amgen/Wyeth$3,657 US mil Procrit/Ep rex Stimulation of red blood cells for treatment of anemia in HIV patients Johnson&Johnson/Ortho Biotech $ 3,324 US mil Aranesp Stimulation of red blood cells for treatment of anemia associated with chronic renal failure Amgen$ 3,273 US mil Rituxan Leukemia and non- Hodgins lymphoma Genentech/Roche/Biogen$ 3,154 US mil Remicade Rheumatoid arthritis and Crohn’s disease Johnson&Johnson/Centoco r/Schering Plough $ 2,535 US mil

36. Epogen Stimulation of red blood cells for treatment of anemia associated with chronic renal failure Amgen$ 2,455 US mil Neulasta Stimulation of production of white blood cells for protection against chemotherapy complications Amgen$ 2,288 US mil NeoRecor mon/Epog in Stimulation of red blood cells Genentech/Roche/Chugai$ 1,710 US mil Herceptin Breast cancer Genentech/Roche$ 1,629 US mil Avonex MS Biogen IDEC$ 1,543 US mil

37. Monoclonal antibodies

39. New challenges…. Increasingly, bacteria and yeast cannot correctly form the human protein: animal cell culture ex) mouse cells require even more careful treatment Large quantities???

40. Opportunities for you? The Bioproducts industry “needs staff that bridge the key disciplines of biology, chemistry and engineering” Canada’s Innovation Strategy, 2001 Government of Canada Bioproducts Sector Profile

41. Main Issues Types of reactors to provide high oxygen transfer Appropriate cell and medium selection Sterilization and maintaining sterility, no cross contamination In pharma, product purity and quality impedes process change Cells and molecules are sensitive to extreme conditions Growth rate and reaction rates are small Product is usually very dilute Non traditional methods of separation Often batch operations