By: ILA JOSHI M.Sc.

IIIrd Semester Roll No-5461

 Downstream Processing: Any treatment of

broth after completion of fermentation or a bioprocess.

Unit Operations: Individual steps or

operations during the downstream processing that change or separate components.

e.g. centrifugation, filtration, precipitation etc.

 Location of product Concentration of product Physical and chemical properties Use of product Standard of purity Magnitude of bio-hazard Impurities Marketable price

1 2 3 4
5

Centrifugation, Filtration

Product Enrichment Evaporation , precipitation , adsorption , ultrafiltration

Primary Isolation of diff. products Liquid Extraction, Precipitation

Chromatography, Ultrafiltration Final Isolation Crystallization followed by centrifugation or filtration

Three major membrane separation processes

Microfiltration Ultrafiltration Reverse Osmosis

Removes contaminants from fluids (liquid or gas). External pressure is not used.

Pore Size: 0.1 to 10 m.

Efficiency is limited up to feed concentration of 23%. Separation of cells, beads from broth As an aid to ultrafiltration or reverse osmosis

A process that retains solutes of higher molecular weight Solvent and solutes are forced under hydraulic pressure (7 atm) through a membrane of very small pore size.(100KDa)

 A wide range of

membranes of polymeric materials is used Pore sizes also can be chosen, as per requirement, from a range of 50-500,000 molecular weight cut off.

Spiral Module
Membrane in consecutive layers inside a tube Increases surface area Cheaper

Tubular membrane
Feed solution flows through membrane core Permeate collected in a tube Expensive

Hollow fiber Membrane

Small (0.6 to 2 mm) diameter tubes or fibers Feed flows through core of fibers Permeate is collected in surrounding cartridge

1. 2. 3. 4.

Virus recovery (10-30KDa) Enzymes recovery Antibiotics recovery Water purification and waste water treatment
Industrial UF System

Prevents blockage of filter membrane by filter cake. Kieselguhr (diatomaceous earth) is most widely used. It is mixed with initial cell suspension & improves porosity of cake. Expensive & absorbs filterate. Not used in case of biomass production or intracellular product.

Types Of Filters
Batch filters
Plate & frame filters
Continuous filters

Pressure leaf filters

Vertical metal leaf filters Horizontal metal leaf filters

Rotary vacuum filters

Stacked disc filters

In other methods the filtration was perpendicular to the membrane. More clogging and wedging of pores. Tangential flow provides: Efficient sepration, > 99.9% cell retention Closed system; for the containment of organisms with no aerosol formation Separation is independent of cell and media densities, in contrast to centrifugation. No addition of filter aid

SCHEMATIC DIAGRAM OF CROSSFLOW FILTRATION.

HPTFF ASSEMBLY

Solvent molecules are forced by an applied pressure to flow through a semi-permeable membrane. Flow of solvent is in direction opposite to one direction dictated by osmotic force, hence the name reverse osmosis. It is used for concentration of smaller molecules than is possible by ultrafiltration

Concentration of fruit juices. 2. Production of whey protein powders. 3. In wine industry. 4. Water purifiers.
1.

Downstream processing of proteins methods and protocols -Mohamed A. Desai Principles of Fermentation Technology- P.F. Stanbury, A Whittaker and S. J. Hall www.wikipedia.org