Introduction Stages of Biotech Ancient Classical Modern

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  • Introduction

  • Stages of BiotechAncientClassicalModern

  • Ancient BiotechBegins with early civilizationDevelopments in agriculture and food productionFew records existUse microorganisms

  • Classical BiotechFollows ancientMakes wide spread use of methods from ancient, especially fermentationMethods adapted to industrial production

  • Era of microorganism (19 century 1944)18th First vaccination in Europe (cowpox), heat sterilization of food and organic chemistry1857. Pasteur :microorganism1897. Germany :Buchner :enzyme A primary metabolism product:ethanol, citric acid B: anaerobic fermentation Commercial production of citric acid1940s Production of penicillin by fermentation

    Most of amino acid isolated 1860-1890

    Enzyme Engineering

  • 2. Enzyme Engineering1953. Grubhofer and Schleith immobilization of enzyme

    1969. Japan :application of immobilized enzyme in industry

    Amino acid production 1976 Genentech first specialist biotech company

  • Modern BiotechManipulation of genetic material within organismsBased on genetics and the use of microscopy, biochemical methods, related sciences and technologies

  • 3. Genetic Engineering 1974. US Boyer and Cohen recombinant DNA1976. first biotechnology company Genentech was established1977. Boyer hGH1986 First rDNA vaccine approve1995 First bacterial genome sequenced2000 Human genome sequenced

  • Application of BiotechnologyFood industry




    IndustryScaleDownstream complexityBiocatalystProductsBiotech market shareBasic chemicalsFine chemicalsDetergentsHealth care/ cosmeticsPharma conventional biopharma

    Food/feedMetal miningWaste treatment Very large




    Very largeVery largeVery largeLow






    MOMO/enzymes/mammalian cells

    MOMammalian cells, MO

    MO/enzymesMO MOOrganic small moleculesOrganic small moleculesEnzymesProteins & small molecules

    Organic small moleculesProteins

    ProteinsMetalsPurified waterVery low




    MediumVery Lowhigh

  • Modeling and Assessment inProcess Development

  • Process design and developmentModeling and simulationSustainabilityassessmentStopProcess conceptIndustrial applicationLiteraturePatentsExpertknowledgeEco-efficientNot eco-efficientImprovementneededWhy must modeling and simulation?Close collaboration with the process designAdditional information from patents, literature, and other external sourcesSimulation used to evaluate the process and guide the R&D to the overall aimRepeated iteratively

  • To gain an understanding of the actual future productionTo realize competitive industrial processes and decision has to be made based on the cost and potentials of a processTo solve a problem that was previously overlooked rises with the development stageTo give a complete picture of the expected production-scale

  • Development Of Bioprocess

  • The Biotechnical Process

  • Solid

  • Unit Operations:Basic step in production processe.g. sterilization, fermentation, enzymatic reaction, extraction, filtration, crystallizationUnit Procedures:Set of operation that take place sequentially in a piece of equipmente.g. charging of substrate to a fermenter, addition of acid to adjust pH, reaction, transfer of fermentation broth to another vessel

  • Upstream processing


    Downstream processing

  • SolidUpstream processingBioreactorDownstream processing

  • Preparation and Storage of Solutions-to provide and store that are needed at some point in the processe.g. preparation of the medium for the bioreactor/buffers in the chromatography-Liquid and solid mixture: filled in tank, mixed by agitation, stored in the tank or transferred to a separate storage tank until is needed in the process-Raw material solutions: prepared with high concentrations to keep the volume of the preparation tanks small-Carbon and nitrogen sources: prepared in separate tanks to avoid Maillard or non-enzymatic browning reactions

  • 2. Sterilization of Input Materials-to preclude contamination of the bioreactorFiltration-to sterilize gaseous streams-membrane filters: Pore size- 0.2-0.3m-prefilters used for dust and other particles

    (ii) Heat Sterilization-heated by steam -cooling water to bring the temperature back to normal-Temperature: 121 C (batch),140-45 C (continuous), holding time: 10-20 min-For continuous: required heat exchanger

    Upstream processing

  • 3. Inoculum preparation-to provide a sufficient amount of active cell to inoculate the production fermenter

    Cleaning-in-Place (CIP)-to prepare the equipment for the next cycle or batchUpstream processing

  • Bioreactor TypesStirred tank bioreactor

    -most commonly used in bioprocess-depends on the complexity of the bioreaction-air, supplied by a compressor, enter the vessel at the bottom under pressure-jacket and/or internal coils allow heating and cooling

  • (ii) Airlift bioreactorBioreactor -mixing is achieved without mechanical agitation by the convection caused by sparged air-lower energy consumption-used for plant and animal cell culture and for immobilized biocatalysts

  • (iii)Packed-bed and fluidized bed bioreactor

    Bioreactor -The immobilized or particulate biocatalyst is filled in a tube-shaped vessel-medium flows through the column (upwards or downwards)-small particle attrition-high velocity of the liquid phase promotes good mass transfer

  • 2. Unit Procedures(i) Filling and transfer of materials in vessels-to bring materials (liquids, solids) into the bioreactor -to transfer parts or the whole reactor volume to the next operation at the end of the bioreaction-the duration should be specified-filled up to only 70-90% to keep some headspace for foam build-up(ii) Agitation-to achieve and maintain homogeneity-to enable efficient heat transfer-energy consumption depends on the rotational speed of bioreactor, fluid density and viscosityBioreactor

  • (iii) Aeration-provides oxygen to meet the aerobic demand of thecells during fermentation-specified by the gas used (Air, pure O2, pure N2, orair enriched with O2 or CO2) and the aeration rate (0.1 and 2 vol. of gas per volume of solution per minute (vvm)(iv) Heat transfer-to change and control the temperature of the bioreactor-to keep constant while exothermic reactions take place in the fermenter-for heating, heat is transferred from a heat-transfer fluid via a heat- transfer surface to the reactor content


  • -for cooling, heat transferred from the fermentor to the cooling fluid-used steam for heating-heating rate depends on the bioreactor volume, typically at 1.5-3.0 C/min for a 10m3 reactor and 1-2 C/min for a 50 m3 reactor-cooling agent : cooling water (20 C), chilled water (5 C), Freon, glycol, sodium chloride brine, calcium chloride brine(v) Foam control-to control the foam formation from the combination of agitation and aeration with the presence of foam-producing and foam-stabilizing substances


  • (vi) pH control-to control and reach the desired pH-the medium is buffered- adjusting and maintained the pH by adding acid or bases (vii) Cleaning-in-place (CIP)-to clean the equipment after every batch


  • Downstream processingBiomass removal-separate the biomass from the fermentation broth-unit operations: centrifugation, microfiltration, rotary vacuum filtration, decanting/sedimentation-depends on a number of parameter (e.g. concentration, particle size, density of biomass,scale operation etc)2. Homogenization/Call Disruption-to break open the cells to release the product into the solution before purification-unit operation: high pressure homogenization, mechanical bead milling

  • 3. Concentration-to reduce the volume of the product stream that has to be processed-reducing equipment size and energy consumption-three methods available:Partial evaporation of the solvent-solution heated up to vaporize some of the solvent, usually water(b)Filtration-semi-permeable membrane retains the product in the retentate but transfers most of the solvent through the membrane(c)Precipitation-adding a precipitation agent or by changing chemical or physical conditions

    without degradationDownstream processing

  • 4. Phase Separation (i) Centrifugation-Used for biomass removal and solid separation-based on density between solid particles and a solution between two immiscible liquids-sedimentation force is amplified by the particle or drop size in centrifugal field in the centrifuge-pretreatment is necessary to increase particle size-maximum throughput defined by the sigma factor and the settling velocity

    Downstream processing

  • (ii) Filtration -to separate particles or large molecules from a suspension or solution-semi-permeable membrane splits the components according to their size-microfiltration:Pore sizes of 0.1-10 mFlux rate: 20 and 250 L/m2-ultrafiltration-Pore sizes of 0.001-0.1 mFlux rate: 20 and 200 L/m2

    Downstream processing

  • -dead-end filtration:(a)particles are retained as a cake through which solvent must pass(b)The pressure drop increases with solids accumulation-cross-flow filtration:(a)The feed is moved tangentially along the membrane to reduce concentration polarization or filter-cake thickness and associatedpressure drop(b) Particles are obtained as concentrated slurry-rotary vacuum filtration:Used only for large-scale filtration with