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1. Large-scale Enzyme Production Tina C. Lung
Dept. of Chemical Engineering and Material Science
CEN 551-Biochemical Engineering
January 22, 2004
2. Examples of Enzymes Protease (subtilisin, rennet)
Hydrolases (pectinase, lipase, lactase)
Isomerases (glucose isomerase)
Oxidases (glucose oxidase)
Produced using overproducing strains of certain organisms.
3. Separation and Purification Disruption of cells
Removal of debris and nucleic acids
Precipitation of proteins
Ultrafiltration of the desired enzyme
4. Separation and Purification The process depends on whether or not the enzyme is intra or extracellular.
Sometimes inactive (dead or resting) cells are used with desired enzyme activity in immobilized form.
No separation and/or purification steps
5. Growing Enzymes (1) Cultivate the organisms producing the desired enzymes.
Production can be regulated
Fermentation conditions ca be optimized for overproduction.
6. Enzymes and Sources Proteases
Overproducing strains of Bacillus, Aspergillus, Rhizopus, and Mucor.
Yeast and Aspergillus.
Certain strains of yeast and fungi.
Flavobecterium arborescens or Bacillus coagulans
7. Growing Enzymes (2) (2) Cell separated from the media usually by filtration or something by centrfugation.
Depending on intra/extracellular nature of the enzyme, the cell or fermentation broth is further processed.
Recovery of intracellular enzymes is more complicated and involves the disruption of cells and removal of debris and nucleic acids.
Increasing permeability of cell membrane (CaCl2 (salt) or dimethylsulfoxide (DMSO) or change in pH
Last resort is cell disruption.
8. Figure 3.23
9. Medical and Industrial Utilization of Enzymes Pharmaceutical Uses
Wants chirally pure compounds (IMPORTANT!).
Often a particular enantiomer maybe useful while others cause side effects or no effect at all.
10. Technological advances
Increasing in wider ranges of process conditions.
Enzymes grow in unusual environments (eg: deep ocean, salt lakes, and hot springs).
New enzymes and better control allow the use of enzymes in more extreme environments.
11. Economics Number of enzymes made at high volume for industrial purposes evolves more slowly.
1996: sales of industrial enzymes--$372 million.
2006: expected to reach $686 million.
12. Table 3.5
13. Industrial Protease Hydrolyze proteins into small peptide groups.
Molds (Aspergillus, Rhizopus, and Mucor)
14. Major Uses Cheese making (rennet)
Meat tenderization (papain, trypsin)
Brewing (trypsin, pepsin)
Detergents (subtilisin Carlsberg)
Medical treatments of wounds
15. Hebei Shenzhou Animal Medicine Co., Ltd Specialized manufacturer of zinc bacitracin in China.
16. Genencor International Second largest developer and manufacturer of industrial enzymes.
Introduced the first industrial scale, recombinant enzyme in 1988.
Leader in the areas of protein engineering, expression/secretion technology and enzyme-substrate interaction.
17. Novozymes Novozymes.com
Largest producers of enzymes.
World Headquarter: Denmark (1941).
More than 500 enzymes in over 130 countries.
18. Words of Wisdom I imagine a future where our biological solutions create the necessary balance between better business, cleaner environment, and better lives.
~Steen Riisgaard, President and CEO