after fermentation which steps done to purify, concentrate and packaging of product

1. Down Stream Processing
Saba Naeem 2015-m.phil-2368

2. Introduction
• For the formation of product we follow that general reaction in which we
provide the substrate and reaction environment for the formation of
Product
• Substrate M.O Product
• For the processing of Product we need three basic steps which are following
• Up stream processing
• Fermentation
• Down stream processing

3. Up stream processing
Upstream processing involves all the steps related with:
 inoculum development
 media development
 improvement of inoculum by genetic engineering process
 optimization of growth kinetics so that product development can improve
tremendously

4. Downstream processing (DSP)
• The various stages of processing that occur after the
completion of the fermentation
• Downstream Processing comprises all operations required
for extraction and purification of a product produced by a
biotechnological process such as microbial fermentation
• It is actually deal with the product

5. Products
• We got three forms of product after fermentation. These forms are following:
• A cell (yeast)
• Extracellular products (a product which after production release from the cell
in to media)
• Intracellular products (a product which exits in cell)

6. Major steps of downstream processing
• There are three major steps which we deal with during downstream
processing
• Purification/ separation
• Concentration
• Packaging

7. Cell product purification
• As we know we have not pure form of product so we need downstream processing to
sale it out in market. The fermenter have:
 Cells
 Substrate
 Product + by-product in it
• So when we talk about cell as product so we do Separation to separate or purify our
product
• For separation we adopt the following two processes
 Centrifugation
 Filtration techniques
 So by centrifugation and filtration we got our product out and rest of things are remain
in solution which are waste for us

8. Extracellular product Purification
• Purification of extracellular product we first need similar process as in case of
cell product
 Centrifugation
 Ultra filtration
• Discard the solid portion and use the liquid portion because it contain our
product of interest but it is not fully purified because it has substrate and by-
product also. So for further purification we do
 Chromatography (GC, HPLC, ion exchange chromatography or other)
 Sedimentation
 Keep in mind that for chromatography and sedimentation we must know the
physical and chemical properties of our product of interest

9. Intracellular product purification
• When product is intracellular so product is in the cell so do centrifugation and
filtration but this time waste the liquid/ supernatant and use the solid part
• Now this solid part has our product inside of it so firstly we need to creak the
cell and for creaking we need to follow physical and chemical techniques
such as:
 Homogenization
 Sonication
 Osmolytic gents
 Now our product came outside and solution contain cell debris and product
so again for purification we do centrifugation and filtration
 Solid discard and store supernatant because it is our product

10. Concentration of product
• As above we discuss different purification techniques to get purified product
but that product is in liquid form so we need to concentrate it
• For concentration we need to follow these process
• Chromatography
• Crystallization
• Precipitation
• Evaporation

11. Packaging of product
• End with packaging of the product in a form that is stable, easily
transportable and convenient.
 Crystallization
 Desiccation
 Lyophilization (dry freezing)
 Spray drying
• May include Sterilization of the product Remove or deactivate trace
contaminants which might compromise product safety viruses or

12. Why downstream processing
•Reduction in bulk
•Concentration enrichment
•Removal of specific impurities (e.g., toxins from therapeutic products)
•Prevention of catalysis other than the type desired (for enzymes)
•Recommended product specifications (e.g., pharmaceuticals
requirement)
•Enhancement of protein stability
•Reduction of protein degradation (e.g. by proteolysis)