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Large-scale Enzyme Production. Tina C. Lung Syracuse University Dept. of Chemical Engineering and Material Science CEN 551-Biochemical Engineering January 22, 2004. Examples of Enzymes. Protease (subtilisin, rennet) Hydrolases (pectinase, lipase, lactase) Isomerases (glucose isomerase)

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large scale enzyme production

Large-scale Enzyme Production

Tina C. Lung

Syracuse University

Dept. of Chemical Engineering and Material Science

CEN 551-Biochemical Engineering

January 22, 2004

examples of enzymes
Examples of Enzymes
  • Protease (subtilisin, rennet)
  • Hydrolases (pectinase, lipase, lactase)
  • Isomerases (glucose isomerase)
  • Oxidases (glucose oxidase)
  • Produced using overproducing strains of certain organisms.
separation and purification
Separation and Purification
  • Disruption of cells
  • Removal of debris and nucleic acids
  • Precipitation of proteins
  • Ultrafiltration of the desired enzyme
  • Chromatographic separation
  • Crystallization
  • Drying
separation and purification4
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
    • Reduces costs
growing enzymes
Growing Enzymes
  • (1) Cultivate the organisms producing the desired enzymes.
    • Production can be regulated
    • Fermentation conditions ca be optimized for overproduction.
enzymes and sources
Enzymes and Sources
  • Proteases
    • Overproducing strains of Bacillus, Aspergillus, Rhizopus, and Mucor.
  • Pectinases
    • Aspergillus niger.
  • Lactases
    • Yeast and Aspergillus.
  • Lipases
    • Certain strains of yeast and fungi.
  • Glucose isomerase
    • Flavobecterium arborescens or Bacillus coagulans
growing enzymes 2
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.
medical and industrial utilization of enzymes
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.

Medical and Industrial Utilization of Enzymes

  • 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.
  • 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.
table 3 5
Table 3.5

Table 3.6

industrial protease
Industrial Protease
  • Hydrolyze proteins into small peptide groups.
  • Obtained from:
    • Bacteria (Bacillus)
    • Molds (Aspergillus, Rhizopus, and Mucor)
    • Animal pancreas
    • Plants
major uses
Major Uses
  • Cheese making (rennet)
  • Baking
  • Meat tenderization (papain, trypsin)
  • Brewing (trypsin, pepsin)
  • Detergents (subtilisin Carlsberg)
  • Tanning products
  • Medical treatments of wounds
hebei shenzhou animal medicine co ltd
Hebei Shenzhou Animal Medicine Co., Ltd
  • Specialized manufacturer of zinc bacitracin in China.
genencor international
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.
  • Largest producers of enzymes.
  • World Headquarter: Denmark (1941).
  • More than 500 enzymes in over 130 countries.
words of wisdom
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