Upstream and Downstream process.pptx.pptx

upstream and downstream process

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

3. Fermentation is a metabolic process Sugar Convert Acid Gas Alcohol

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. 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

6. 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

7. 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

8. 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

9. 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

10. 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

11. 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

12. 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

13. Solid-liquid separation or clarification Primary operation Separation of whole cells Removal of cell debris Collection of protein precipitate Collection of inclusion bodies etc.,

14. 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

15. 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

16. 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

17. 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

18. 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, p ᴴ, molecular size, protein, bio-specific binding reactions etc.,

19. 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

20. Stages in Downstream Processing

21. 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.

22. 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.

23. Filter media Two main types of filter media are solid sieve - which traps the solid particles bed of granular materials - retains the solid particles

24. 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

25. 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.

26. 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.

27. 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.

28. Product Isolation reducing the volume of material to be handled and concentrating the product. the unit operations involved Solvent extraction ultra filtration precipitation

29. 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 .

30. 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

31. 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.

32. 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

33. Products recoverable using downstream process are : Antibiotics Vitamins Enzymes Pharmaceutical products Food products Dairy products Drugs

34. video https://www.youtube.com/watch?v=9GVY2tWRNxw&t=39s

35. 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

36. Thank you