logging in or signing up FERMENTATION-DIFFERENT STAGES jeejonu Download Post to : URL : Related Presentations : Let's Connect Share Add to Flag Embed Email Send to Blogs and Networks Add to Channel Copy embed code: Embed: Flash iPad Dynamic Copy Does not support media & animations Automatically changes to Flash or non-Flash embed WordPress Embed Customize Embed URL: Copy Thumbnail: Copy The presentation is successfully added In Your Favorites. Views: 1183 Category: Education License: All Rights Reserved Like it (0) Dislike it (0) Added: July 20, 2012 This Presentation is Public Favorites: 0 Presentation Description THE SLIDE INCLUDES DIFFERENT STAGES INVOLVED IN FERMENTATION PROCESS.............HAVE A LOOK... Comments Posting comment... Premium member Presentation Transcript FERMENTATION (Different Stages Of Operation) : FERMENTATION (Different Stages Of Operation) Do what you can, with what you have, where you are. Theodore Roosevelt 1 JEEJO N.U M.PHARM PHARMACEUTICS email@example.comDISCUSSION ON:: DISCUSSION ON: MAINTENANCE OF CULTURE INOCULUM PRODUCTION DOWNSTREAM PROCESSING FERMENTATION SCALE-UP 2MAINTENANCE OF CULTURE: MAINTENANCE OF CULTURE To maintain the physiological properties of high yielding strains. Agar Slope Under Paraffin Deep Freezing Lyophilization 3INOCULUM PRODUCTION: INOCULUM PRODUCTION Most critical stage in any fermentation process. The size of the inoculum is between 5-10% of fermentation process. In each stage rigorous control is necessary to avoid both mutation and contamination. 4DOWNSTREAM PROCESSING-MULTISTAGE OPERATION : DOWNSTREAM PROCESSING-MULTISTAGE OPERATION ‘The isolation and purification of a biotechnological product to a form suitable for its intended use’. Depends on the nature of end product, its concentration, stability and degree of purification required. Product recovery yield is higher if the number of steps in DSP is lower. 5STAGES IN DSP: STAGES IN DSP Solid-liquid separation/clarification Release of intracellular components Extraction Concentration Purification Drying Formulation. 6(I) SOLID-LIQUID SEPARATION/CLARIFICATION: (I) SOLID-LIQUID SEPARATION/CLARIFICATION FLOTATION FLOCCULATION FILTRATION CENTRIFUGATION 7FLOTATION:: FLOTATION: Particles are adsorbed on gas bubbles, get trapped in a foam layer & collected. 8FLOCCULATION:: FLOCCULATION: The cells form large aggregates to settle down for easy removal. Flocculating agents : Inorganic salts, Organic polyelectrolyte's, Mineral hydrocolloid’s. 9FILTRATION: FILTRATION For separating the biomass and culture filtrate. Efficiency of filtration depends on: Size of organism Presence of other organisms Viscosity of the medium Temperature 10 FILTRATION EQUIPMENTS:: FILTRATION EQUIPMENTS: ROTARY DRUM VACUUM FILTER 11PowerPoint Presentation: 12 FILTER PRESSPowerPoint Presentation: MEMBRANE FILTER PRESS 13CENTRIFUGATION: : CENTRIFUGATION: Removal of microbial cells and other discrete large particles. Batch centrifugation Continuous flow centrifugation Separation of particulate debris is inefficient by centrifugation. 14 CENTRIFUGES: : CENTRIFUGES: TUBULAR BOWL TYPE CENTRIFUGE 15PowerPoint Presentation: MULTICHAMBER BOWL TYPE CENTRIFUGE 16PowerPoint Presentation: DISC STACK CENTRIFUGE 17PowerPoint Presentation: THE DECANTER OR SCROLL CENTRIFUGE 18PROPERTIES OF INDUSTRIAL CENTRIFUGES: PROPERTIES OF INDUSTRIAL CENTRIFUGES Tube High centrifugal force Good dewatering Easy to clean Chamber Large solids capacity Good dewatering Bowl cooling possible Disc type Solids discharge No foaming Bowl cooling possible Limited solids capacity Foams Difficult to recover protein No solids discharge Cleaning difficult Solids recovery difficult Poor dewatering Difficult to cleanCentrifugation properties of different cell types: Centrifugation properties of different cell types Bacteria Small cell size Resilient Yeast cells Large cells Resilient Filamentous fungi Mycelial Resilient Cultured animal cells Large cells Very fragile High speed required Low cell damage Lower speed required Low cell damage Lower speed required High water retention in pellet Very susceptible to damage(II) RELEASE OF INTRACELLULAR COMPONENTS : 21 (II) RELEASE OF INTRACELLULAR COMPONENTS PHYSICAL METHODS CHEMICAL METHODS ENZYMATIC METHODS Ultrasonication Alkalies Lysozyme Osmotic shock Organic solvents Glucanase , mannanase & protease. Thermolysis Detergents High-pressure homogenization Impingement Grinding with glass beads CELL DISRUPTION(III)EXTRACTION:: (III) EXTRACTION: The process of removing a compound or a group of compounds from a mixture or from cells into a solvent phase is called extraction. Liquid – liquid extraction Whole broth (medium + cells) extraction Aqueous multiphase extraction 22(IV)CONCENTRATION: (IV) CONCENTRATION Evaporation Liquid-liquid extraction Membrane filtration Precipitation Adsorption 23EVAPORATORS: 24 EVAPORATORS PLATE EVAPORATORPowerPoint Presentation: 25 FALLING FILM EVAPORATORPowerPoint Presentation: 26 THIN FILM EVAPORATORPowerPoint Presentation: 27 CENTRIFUGAL TYPE THIN-FILM EVAPORATORLIQUID-LIQUID EXTRACTION: LIQUID-LIQUID EXTRACTION “ Transferring the solute from one liquid phase to another liquid phase” Extraction of low MW products. Extraction of high MW products. 28PowerPoint Presentation: 29 EXTRACTION OF LOW MW PRODUCTS. EXTRACTION OF HIGH MW PRODUCTS. PHYSICAL EXTRACTION AQUEOUS TWO-PHASE SYSTEMS (ATPS) DISSOCIATIVE EXTRACTION REVERSE MICELLER SYSTEMS REACTIVE EXTRACTION SUPERCRITICAL FLUID EXTRACTION (SCF)MEMBRANE FILTRATION: MEMBRANE FILTRATION Ion exchange resins Adsorption resins-styrene, sulfoxide,acrylic ester. Pervaporation - membrane filtration coupled with evaporation. Perstraction - membrane filtration coupled with solvent extraction. 30PowerPoint Presentation: TYPE PARTICLE SIZE COMPOUND OR PARTICLE SEPARATED MICROFILTRATION 0.1 – 10 μ m Cells or cell fractions, Viruses. ULTRAFILTRATION 0.001- 0.1 μ m Compounds with MW > 1000 (e.g. enzymes). HYPERFILTRATION 0.0001- 0.001 μ m Compounds with MW < 1000 (e.g. lactose). Major Types Of Filtration Processes 31PRECIPITATION: PRECIPITATION Separation technique used for concentration of proteins and polysaccharides. Constitutes the only unit operation in the recovery of bulk proteins. 32PowerPoint Presentation: 33 MODE EXAMPLE Addition of neutral salts (NH 4 ) 2 SO 4 Addition of organic solvents Ethanol, acetone , Propanol Addition of non-ionic polymer PEG Addition of charged polymer Polyacrylic acid , Polyethyleneimine Increase in temperature change in pH MODES OF PROTEIN PRECIPITATIONADSORPTION: ADSORPTION Activated charcoal. Cellulose based adsorbents- for protein precipitation. Polystyrene,methacrylate and acrylate based adsorbents- for low molecular weight compounds . 34(V) PURIFICATION: (V) PURIFICATION CRYSTALLIZATION CHROMATOGRAPHIC METHODS 1. Adsorption chromatography 2.Ion exchange chromatography 3.Hydrophobic interaction chromatography 4. Affinity chromatography 5.Size exclusion chromatography 6.Radial flow chromatography 7.Perfusion chromatography 8.HPLC 35PowerPoint Presentation: 36 Chromatography Separation principle Size-exclusion (gel filtration) Size & charge Ion-exchange chromatography Net charge Chromatofocusing Net charge Hydrophobic interaction chromatography Hydrophobicity Affinity chromatography Molecular recognition Immobilized metal-ion affinity chromatography Metal ion binding Covalent chromatography Content of free –SH groups. VARIOUS CHROMATOGRAPHIC TECHNIQUES FOR PROTEIN SEPARATIONCHROMATOGRAPHY SYSTEM: CHROMATOGRAPHY SYSTEM Lab Scale Chromatography System Large Scale Chromatography System(VI) DRYING:: (VI) DRYING: Makes the products suitable for handling and storage. (i) Vacuum drying (ii) Freeze drying (iii) Spray drying 38PowerPoint Presentation: 39 SOME OF THE FREQUENTLY USED DRYING EQUIPMENTS Type of dryer Mode of heat transfer Movement of the product Belt dryer Convection Intensive due to gas flow Fluidized bed dryer Convection Intensive due to gas flow Spray- dryer Convection Intensive due to gas flow Freeze- dryer Contact and radiation None or mechanical Drum- dryer Contact Slight mechanical Chamber dryer Convection and contact NonePowerPoint Presentation: 40 FREEZE-DRYER SPRAY-DRYER VACUUM -DRYER(VII) FORMULATION: (VII) FORMULATION The commercial viability of a biotechnological product is dependent on the maintenance of its activity and stability during distribution and storage. Stabilizing agents are included in the formulations to prolong the product shelf life . 41PowerPoint Presentation: Media Prep Working Cell Bank Sub- Culture Inoculum Sub- Culture Sub- Culture Sub- Culture Sub- Culture Large Scale Bioreactor Wave Bag Seed Bioreactors Fermentation 150L Bioreactor 750L Bioreactor 5,000L Bioreactor 26,000L Bioreactor Depth Filtration Collection Centrifuge Harvest/Recovery Harvest Collection Tank 1,500L Filter Chromatography Skid Anion Exchange Chromatography (QXL) Column Eluate Hold Tank 8,000L Eluate Hold Tank 6,000L Filter Chromatography Skid Protein A Chromatography Column Chromatography Skid Column Eluate Hold Tank 20,000L Hydrophobic Interaction Chromatography (HIC) Eluate Hold Tank 20,000L Viral Inactivation Eluate Hold Tank 5,000L Filter Chromatography Skid Anion Exchange Chromatography (QFF - Fast Flow) Column Post-viral Hold Vessel 3,000L Viral Filtering Ultra Filtration Diafiltration Bulk Fill Purification 24 days 31 days 8 days 1 day Upstream Downstream Process OverviewFERMENTATION SCALE-UP: FERMENTATION SCALE-UP ‘ The phenomenon of developing industrial fermentation process in stages ’. To develop optimal environmental and operating conditions at different levels. 43REFERENCES: REFERENCES Colin Ratledge & Bjorn Kristiansen, Basic Biotechnology, Cambridge University Press, 151-211. Vinita Kale & Kishore Busari, Applied Microbiology, Himalaya Publishing House, First edition-2001, 163-202,303-330. Biotechnology by B D Singh, Kalyani publishers, First edition- 1998. Giriraj Kulkarni T, Biotechnology And its applications in pharmacy, jaypee publications, first edition 2002; page no:73-99. S S Kori & M A Halki, Pharmaceutical Biotechnology-Fundamentals and applications, page no: 135-156. Dr. U. Satyanarayana, Biotechnology, First edition-2005, Books and Allied pvt.ltd, page no:268-280. 44PowerPoint Presentation: 45 You do not have the permission to view this presentation. In order to view it, please contact the author of the presentation.