chapter 3 large-scale enzyme production

8/3/2019 Chapter 3 Large-Scale Enzyme Production

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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

chapter 3 large-scale enzyme production

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