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PURIFICATION OF
FERMENTATION
PRODUCTS
Usman Yousaf
Introduction to Purification
Many biological processes
require a purification scheme to
reduce the fermentation broth to
its pure final product. Once
product is made in the
production fermenter, the broth
is still highly contaminated.
Precipitation
Precipitation is a procedure where the addition of a ionic solution
to an ionic fermentation broth forms insoluble particles, where
the desired product is usually contained within those particles.
Ionic fermentation broths usually consist of enzymes or proteins.
The ways to precipitate out a product can vary from simple pH
and temperature changes to chemical reactions involving metal
ions. Precipitation reactions are carried out in reactors,
continuous and batch.
Precipitation
Typical agents used in precipitation to render the compound of
interest insoluble, and these include:
Acids and bases
Salts such as ammonium and sodium sulphate
Organic solvents
Non-ionic polymers
Protein binding dyes
Filtration
Filters use a filter cloth or some porous material along with
applied pressure to push smaller particles through the filter, thus
separating elements of the solution based on size. Filtration for
biological materials is generally completed using batch filtration,
rotary drum filtration, or ultra filtration methods e.g..,
Batch Filtration
Rotary Drum Filtration
Ultra filtration
Centrifugation
Centrifugation involves separation of liquids and particles
based on density. Centrifugation can be used to separate cells
from a culture liquid, cell debris from a broth, and a group of
precipitates
Centrifugation
Although a centrifuge may be expensive when compared with a filter it
may be essential when:
Cell disruption
oMicro-organisms are protected by extremely tough cell walls. In order to release their
cellular contents a number of methods for cell disintegration
Physico-mechanical Method:
Liquid Shear
Solid Shear
Agitation with abrasives
Freeze-thawing
Ultrasonication
Chemical Methods:
Detergents
Osmotic shock
Alkali Treatment
Cell Disruption
Liquid Shear
Operation:
Cell Disruption
Freeze Thawing:
Ball Mill
Solid Frozen cell paste, cells attached to or within a solid
matrix.
Large scale
http://www.unitednuclear.com/mills.htm
Cell Disruption
Challenge: Damage to the product
- Heat denaturation
- Oxidation of the product
- Unhindered release of all intracellular products