1. UPSTREAM
AND
DOWNSTREAM PROCESS
Submitted by:-
A.Anbu Abubakkar Sidik
II MSc Microbiology
PG & Research Department of Microbiology
SRI PARAMAKALYANI COLLEGE
REACCREDITED WITH B GRADE WITH A CGPA OF 2.71 IN THE SECOND CYCLE OF NAAC
AFFILIATED TO MANOMANIUM SUNDARANAR UNIVERSITY, TIRUNELVELI.
ALWARKURICHI 627 412, TAMIL BADU, INDIA
(PG & RESEARCH DEPARTMENT OF MICROBIOGY)
GOVERNMENT AIDED
Submitted to:-
Dr.Viswanathan
Head of the Department
PG & Research Department of Microbiology
SEMESTER- IV
2. HISTORY
• It is known and practiced by humankind prehistoric
times since
• The production of beer, bread, wine etc., are established
in ancient Egypt
• In 1857 Pasteur developed the fermentation process
• In 1897 Edward Buchner discovered that sucrose could
be fermented to alcohol by yeast
• In 1984 first constructed bioreactor by Stanberry and
whitaker
Louis Pasteur
4. Upstream bioprocessing is
the first part of a bioprocess
that involves steps of product
development
Downstream bioprocessing is
the second part of a
bioprocess that involves that
involves steps of product
harvest
Isolation and selection of the
microorganisms development
of the inoculum. Media
preparation, inoculation and
incubation
Extraction, purification,
quality management and
packaging of the product
Product development Harvesting of the product
Identification and selection
of desired microorganisms
Upstream process
Upstream process Downstream process
Definition
Main step
involved
Major event
Followed by
5.
6.
7.
8. UPSTREAM PROCESS
The pre-fermentation stage
• Isolation
• Improvement
• Producing of microorganisms
• Screening method: isolate microbes to produce desired
products
Two methods
• Primary screening checking the quality of microbes done in
agar plates
• Secondary screening checking the quantitative of microbes
done in liquid media
2 methods
Primary
screening
Secondary
screening
9. Microbes isolated from natural sources thus is improved to get productive strains by
using
• Recombination
• Mutation
• Cell fusion
• Gene cloning
Media formulation : growth medium must have essential nutrients for microbial growth
for successful fermentation process
Two kind media
• Inoculum media: enrich the culture
• Production media : contain carbon and nitrogen
Raw materials : corn molasses, cellulose, corn, Streep liquor soybean, sugar, beet
molasses, malt extract etc..
Inoculum media
Production media
10. Upstream processing includes formulation of the fermentation
medium, sterilization of air, fermentation medium and the fermenter,
inoculum preparation and inoculation of the medium.
The fermentation medium should contain an energy source, a carbon
source, a nitrogen source and micronutrients required for the growth
of the microorganism along with water and oxygen, if necessary
A medium which is used for a large scale fermentation, in order to
ensure the sustainability of the operation, should have the following
characteristics;
It should cheap and easily available
It should maximize the growth of the microorganisms, productivity
and the rate of formation of the desired product
It should minimize the formation of undesired products
11. • Usually waste products for other industrial processes, such as molasses,
lignocelluloses wastes, cheese whey and corn steep liquor, after modifying with
the incorporation of additional nutrients, are used as the substrate for many
industrial fermentation.
• Sterilisation is essential for preventing the contamination with any undesired
microorganisms.
• Air is sterilised by membrane filtration while the medium is usually heat
sterilised
• Any nutrient component which is heat labile is filter- sterilised and later added
to the sterilised medium.
• The fermenter may be sterilised together with the medium or separately
12. • Inoculum build up is the preparation of the seed culture in amounts
sufficient to be used in the large fermenter vessel.
• This involves growing the microorganisms obtained from the pure
stock culture in several consecutive fermenter
• This process cuts down the time required for the growth of
microorganism in the fermenter, thereby increasing the rate of
productivity
• Then the seed culture obtained through this process is used to
inoculate the fermentation medium
13. Pure culture
under good
condition
Contain seed
culture in suitable
conditions
Containing culture
for large scale
fermentation
Fermenter contain medium and
culture for mass culture contain
suitable conditions
14. DOWNSTREAM PROCESS
• When fermentation is over, the desired product is recovered
from the growth medium.
• Then the product is extraction purification packed of a
biotechnological product from fermentation is referred to as
DSP or product recovery or downstream processing.
• The end products include Antibiotics, Amino acids, Vitamins,
Organic acid, Industrial enzyme, vaccines etc.
• It is complex and important as fermentation process
15.
16.
17.
18. CELL-DISRUPTION OR DISINTEGRATION
• Mechanical methods:
• Shear forces in solid matter and solution
• Non mechanical methods: Lysis
• Physical- freezing and thawing: non high osmotic pressure, shock
• Chemical- surface active agents, solvents, antibiotics etc.,
• Enzymatic – lysozyme
• Drying
• Freeze-drying, air drying, pressure release, drying with solvents
19.
20.
21. SOLID-LIQUID SEPARATION OR
CLARIFICATION
• Primary operation
• Separation of whole cells
• Removal of cell debris
• Collection of protein precipitate
• Collection of inclusion bodies etc.,
22. SOLID-LIQUID SEPARATION
(CLARIFICATION)
• Coagulation
• Colloids into small flocs using simple electrolytes
• Flocculation
• Agglomeration of these small flocs into larger settle – able particles using
polyelectrolytes.
• Flotation
• Enrichment of microorganisms
• Filtration
• The separation of suspended particles from liquid
• Centrifugation
• Gravitational force used for separation the particles
23. CONCENTRATION METHODS
• The purity or concentration of metabolite
• Evaporation – steam as heat source
• Extraction - the cell mass and more or less clear solution obtained
• Adsorption- special polymer resins (chemical) used for the isolation
of hydrophilic metabolites that cannot be extract with organic
solvents
• Filtration – separation of biomolecules and particles ( pore size)
• Precipitation – removal of product from the solvent
• Dialysis – semipermeable membrane
24. PURIFICATION
• Purify relatively low concentration of metabolic products
• The chromatography technique are used
• Purification is a main process in fermentation
• Desired product purification is important
• Many techniques are used for the purification
25. FORMULATION
• Products is depended on the maintenance of its activity and
stability during distribution and storage
• Concentration solution after removing most of the wate
• Drying
• Withdrawal of water from the products
• Crystallization
• Established method used for concentration of the bio-
products
• Widely used for final purification
26. MONITORING OF DSP
• To keep control over the presence and concentration of
target molecule
• Monitored by using sensor
• Mostly used in pharmaceuticals
• Monitor the UV- absorbance, conductivity, pᴴ, molecular size,
protein, bio-specific binding reactions etc.,
27. MARKETING
• The formulated product is packed and sent to the market
for the consumers.
• The purified product is modified before marketing
• Product may be in aqueous form or in solid form
• The nature of the product have 98-100% in pure for seal in
market
• The purity of the final product depends upon the successful
operation of all these downstream processes
29. REMOVAL OF INSOLUBLE'S
•
• capture of the product as a solute in a particulate-free
liquid
Example
• separation of cells, cell debris or other particulate
matter from fermentation broth containing an
antibiotic.
30. TYPICAL OPERATIONS
Filtration
• A mechanical operation used for the separation of solids
from fluids (liquids or gases) by interposing a medium to
fluid flow through which the fluid can pass, but the solids in
the fluid are retained.
31. FILTER MEDIA
Two main types of filter media are
• solid sieve - which traps the solid particles
• bed of granular materials - retains the solid
particles
32. Points to be considered while selecting the Filter
media:
ability to build the solid.
minimum resistance to flow the filtrate.
resistance to chemical attack.
minimum cost.
long life
33. CENTRIFUGATION
• use of the centrifugal force for the separation of
mixtures
• More-dense components migrate away from the axis
of the centrifuge
• less-dense components migrate towards the axis.
34. FLOCCULATION
• process where a solute comes out of solution in the form of flocs or
flakes. Particles finer than 0.1 μm in water remain continuously in
motion due to electrostatic charge which causes them to repel each
other
• Once their electrostatic charge is neutralized (use of coagulant) the
finer particles start to collide and combine together .
• These larger and heavier particles are called flocs.
35. LYOPHILIZATION
• freezing the material
• reducing the surrounding pressure and adding
enough heat to allow the frozen water in the
material to sublime directly from the solid phase to
gas.
36. PRODUCT ISOLATION
• reducing the volume of material to be handled and
concentrating the product.
• the unit operations involved
Solvent extraction
ultra filtration
precipitation
37. PRECIPITATION
• formation of a solid in a solution during a chemical
reaction.
• solid formed is called the precipitate and the liquid
remaining above the solid is called the supernate.
38. PRODUCT PURIFICATION
• To separate contaminants that
resemble the product very closely in
physical and chemical properties.
• Expensive and require sensitive and
sophisticated equipment.
This purification filter cost is
(Rs:20000 ) .so the purification
equipment is sophisticated
39. CRYSTALLIZATION
• process of formation of solid crystals precipitating
from a solution, melt or more rarely deposited
directly from a gas.
• chemical solid-liquid separation technique, in which
mass transfer of a solute from the liquid solution to
a pure solid crystalline phase occurs.
40. PRODUCT POLISHING
• Final processing steps which end with packaging of the
product in a form that is stable, easily transportable and
convenient.
• Crystallization, desiccation, lyophilization and spray drying
are typical unit operations
41. PRODUCTS RECOVERABLE USING
DOWNSTREAM PROCESS ARE :
• Antibiotics
• Vitamins
• Enzymes
• Pharmaceutical products
• Food products
• Dairy products
• Drugs
45. REFFERENCE
• Text book of fermentation technology by H.A.Modi
• Text book of fermentation technology by R.C.Duby
• Text book of fermentation technology by Dr. Praksh S Lohar
• Also net references
