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Large-scale Enzyme Production - Tina C. LungSyracuse UniversityDept. of Che...

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. Disruption of cellsRemoval of debris and nucleic acidsPrecipitation of proteinsUltrafiltration of the desired enzymeChromatographic separationCrystallizationDrying.

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Large-scale Enzyme Production

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Large scale enzyme production l.jpg

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 l.jpg

Examples of Enzymes

  • Protease (subtilisin, rennet)

  • Hydrolases (pectinase, lipase, lactase)

  • Isomerases (glucose isomerase)

  • Oxidases (glucose oxidase)

  • Produced using overproducing strains of certain organisms.

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Separation and Purification

  • Disruption of cells

  • Removal of debris and nucleic acids

  • Precipitation of proteins

  • Ultrafiltration of the desired enzyme

  • Chromatographic separation

  • Crystallization

  • Drying

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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

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Growing Enzymes

  • (1) Cultivate the organisms producing the desired enzymes.

    • Production can be regulated

    • Fermentation conditions ca be optimized for overproduction.

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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

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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.

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Figure 3.23

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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.

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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.

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  • 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.

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Table 3.5

Table 3.6

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Industrial Protease

  • Hydrolyze proteins into small peptide groups.

  • Obtained from:

    • Bacteria (Bacillus)

    • Molds (Aspergillus, Rhizopus, and Mucor)

    • Animal pancreas

    • Plants

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Major Uses

  • Cheese making (rennet)

  • Baking

  • Meat tenderization (papain, trypsin)

  • Brewing (trypsin, pepsin)

  • Detergents (subtilisin Carlsberg)

  • Tanning products

  • Medical treatments of wounds

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Hebei Shenzhou Animal Medicine Co., Ltd

  • Specialized manufacturer of zinc bacitracin in China.

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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.

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  • Largest producers of enzymes.

  • World Headquarter: Denmark (1941).

  • More than 500 enzymes in over 130 countries.

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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