BIOTEHNOLOGY PROCESSING PRINCIPLE What is biotechnology? • The Office of Technology Assessment of the US Congress defined as: “Any techniques that uses living organisms or substances from those organisms, to make or modify a product, to improve plants or animals, or to develop microorganisms for specific uses.” Why Biotechnology Such A Big Deal? • 1. 2. 3. 4. 5. 6. The benefits of biotechnology may be discussed under 6 major categories: Agriculture (crop, livestock). Industry (ethanol, biofuel, bioplastic). Health/medicine (diagnostic tool, vaccine) Environmental (bioremediation, disease resistant cultivar) Forensic (crime, paternity disputes) Advancement of knowledge What is a bioprocess? • An industrial operation in which living systems are used to transform raw materials (biological or nonbiological) into products. • It is a multidiscipline (biochemistry + chemical engineering + biotechnology) area. • Process to use biology system in commercial scales. Biotechnology can be bring into bioprocess because: 1.The present of engineering innovation: - exp. genetic engineering, discovery of sitespecific enzyme cleavage of DNA by restriction endonucleases in bacteria is the key to DNA manipulation. 2. The development of appropriate enabling technologies: - Polymerase chain reaction (PCR) technology - Blotting technology - Fermenter technology 3.Economic opportunity - Arise from identifiable potential benefit that may accrue to society from development of a product. KEY STEPS IN BIOPROSES ENGINEERING 1. Pretreatment 2. Bioreaction 3. Downstream processing pretreatment Suit raw material into process equipment Design of process Equipment According to Physical & chemical properties Environment factorsphysiologically tolerance of the organism and enzymes. Bioreaction 1. Bioreactor fluids 2. Media sterilization 3. Growth media 4. Microbial growth 5. Mirobial culture systems in bioreactors 6. Bioreactors 1. i. ii. Bioreactor Fluids Medium in which a bioprocess occurs is usually a liquid or slurry. two main kinds of biofluids and slurries used. Newtonian fluids Non-Newtonian fluids 2. Media Sterilization To initiate specific microorganisms for a specific purpose. Usually filter sterilization or heat sterilization. Heat sterilization may be accomplished in batch sterilization or continuous sterilization. 3. Growth Media - Formulated to maintain microbial growth for economic production of the target products. - Should consists basic components such as carbon source, nitrogen source, minerals, vitamins, micronutrients, hormones and oxygen. - May contain additives that facilitate the process e.g antifoams. 4. Microbial Growth Progresses through four basic phases (lag, exponential, stationary and death). Usually 5 to 10 percent by volume of microbial suspension in a rapid exponential growth phase may be used. 5. Microbial Culture System in Bioreactors Grown in three basic ways: 1. Batch fermentation 2. Fed-batch fermentation 3. Continuous fermentation. 6. Bioreactors (will be discussed further) Designs Mode of operation Downstream Processing - To retrieve, purify and pack products for use of sales. - Involves a number of processing. 1. Solid-liquid separation - To separate cells (solid) from the liquid. - Commonly used: centrifugation and filtration. 2. Cell disruption - Extracellular product: the cells are discarded and the medium is concentrated and purified. - Intracellular product: cells have to be disrupted using nonmechanical or mechanical method. i. - Nonmechanical methods Using various chemicals, including alkali, organic solvents, and detergents. - Also some enzymes such as lysozyme, EDTA, β-1,3-glucanase, mannase and chitinase. - Other physical methods are osmotic shock and pressure. ii. Mechanical methods - Two catagories: solid shear and liquid shear. - Solid shear: grinding of frozen cells through narrow gaps under high pressure. - Liquid shear: acceleration of the suspension at a high velocity and under high pressure. 3. Precipitation - To convert the soluble protein product to insoluble form. - Commonly used is salting-out technique using anions and cations. 4. Chromatography - To separate components of mixtures based on their differential physiochemical interaction and a stationary phase. - The stationary phase is a solid packed in a column by porous and hydrophilic substances like agarose, cross-linked polyacrylamide, and cellulose. 5. Drying - Required for safe-storage, packaging, and transportation. - Dehydration is used for vaccines, enzymes, pharmaceuticals and others. - For thermolabile products or biologically active, spray-dried or freeze-dried. References: 1. Acquaah, G. (2004). Understanding Biotechnology:An Integrated and CyberBased Approach. Pearson-Prentice Hall. B. Mode of cells growth i. Immobilized cell system ii. Suspension cultures Immobilized cell system Advantages: 1.Provides high cell concentrations 2.Provides cell reuse 3. Eliminates cell washout problems at high dilution rates 4. Allows high volumetric productivities. 5. Provide favorable microenvironmental conditions for cells, resulting in better performance of biocatalytics. 6. In some cases, improves genetic stability 7. Protection against shear damage. Limitation: 1.The interest product should b excreted by the cells. 2.