chapter 3 large-scale enzyme production
TRANSCRIPT
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Large-scale Enzyme
Production
Tina C. Lung
Syracuse UniversityDept. of Chemical Engineering and Material Science
CEN 551-Biochemical Engineering
January 22, 2004
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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 proteinsUltrafiltration of the desired enzyme
Chromatographic separation
CrystallizationDrying
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Separation and Purification
The process depends on whether or notthe enzyme is intra or extracellular.
Sometimes inactive (dead or resting)cells are used with desired enzymeactivity in immobilized form.
No separation and/or purification steps
Reduces costs
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Growing Enzymes
(1) Cultivate the organisms producingthe desired enzymes.
Production can be regulated
Fermentation conditions ca be optimizedfor 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 arborescensor Bacillus coagulans
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Growing Enzymes (2)
(2) Cell separated from the media usually byfiltration or something by centrfugation. Depending on intra/extracellular nature of the
enzyme, the cell or fermentation broth is furtherprocessed.
Recovery of intracellular enzymes is morecomplicated and involves the disruption of cellsand removal of debris and nucleic acids.
Increasing permeability of cell membrane (CaCl2 (salt) ordimethylsulfoxide (DMSO) or change in pH
Last resort is cell disruption.
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Figure 3.23
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Medical and IndustrialUtilization of Enzymes
Pharmaceutical Uses
Wants chirally pure compounds(IMPORTANT!).
Often a particular enantiomer maybe usefulwhile others cause side effects or no effectat all.
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Technological advances Increasing in wider ranges of process
conditions.
Enzymes grow in unusual environments(eg: deep ocean, salt lakes, and hotsprings).
New enzymes and better control allow the
use of enzymes in more extremeenvironments.
Medical and IndustrialUtilization of Enzymes
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Economics
Number of enzymes made at highvolume for industrial purposes evolvesmore slowly.
1996: sales of industrial enzymes--$372million.
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 peptidegroups.
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 productsMedical treatments of wounds
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Hebei Shenzhou AnimalMedicine Co., Ltd
Specialized manufacturer of zincbacitracin in China.
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Genencor International
Second largest developer and manufacturerof industrial enzymes.
Introduced the first industrial scale,recombinant enzyme in 1988.
Leader in the areas of proteinengineering, expression/secretion
technology and enzyme-substrateinteraction.
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Novozymes
Novozymes.com
Largest producers
of enzymes.World
Headquarter:Denmark (1941).
More than 500enzymes in over130 countries.
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Words of Wisdom
I imagine a future where ourbiological solutions create thenecessary balance between betterbusiness, cleaner environment,and better lives.
~Steen Riisgaard, President andCEO