Topic 1: Introduction to Bioproducts and BioseparationsOBJECTIVES I. Bioproducts Classification and characteristics of bioproducts Characteristic of fermentation broth on downstream processing. Broad classification of bioproducts. Primary and secondary metabolite. II. Bioseparations Introduction to bioseparations. Stages of downstream processing. Basic principles of engineering analysis. Criteria for process development. BIOPRODUCTS: Classification of Bioproducts What is bioproducts: Chemical substances made by living things ranging from small molecules to higher molecules (macromolecules). Derived by extraction from original host or by synthesis in bioreactor containing cells or enzymes. They are sold for their chemical activity; Compound Function Methanol Ethanol Solvent Fuel and pharmaceutical use Penicillin Taxol Steptokinase Whole cell Bacillus thuringienisis Antibacterial agent Anticancer Blood dissolving activity Insecticide monoclonal antibodies and insulin) etc.Range of Characteristics of Bioproducts: 3 major categories: 1. factor VIII. Bulk industrial product of relatively low purity (antibiotic. . amino acids. Very high value. High value. interferone. human growth hormon. 3. organic acids. etc. produced in tens or hundreds kg. produced in the range of grams to kg. of high purity. ethanol. low volume (therapeutic proteins and enzymes. urokinase). 2. low volume and high purity products (diagnostic enzymes. Correlation between market volume or concentration of desired product and selling price or purity requirements: . . metal ions and other chemicals.000.000) Charge distribution Hydrophobicity Structure and immunogenic structure Specific activity Most are fragile. temperature. shear force during stirring and presence of denaturing agents such as surfactants. ionic strength and the nature of solvent. Different separation principles and mechanisms for isolation and purification depends on: Molecular mass (50 to 2. bioactivity and stability affected by factors such as pH. BIOPRODUCTS: Characteristics of Bioproducts . sedimentation. . centrifugation. subsequently pilot plant study. ethanol by yeast and organic acids and enzymes by bacteria. conventional separation via chemical engineering unit operations are used: Unit operations: filtration. In classical biotechnology. adsorption and liquid-liquid extraction are well described mathematically and easy scaled-up. the microorganisms are induced under specific conditions. products produced naturally by the living cells as of penincillin by molds. In modern biotechnology. The type of microorganisms and their morphological features (size and shape) size for microbial. but broth is lower 110 kg/m3 (70-80% water) . dried biomass density of 1400 kg/m3. plant and animal cell and their agglomerates bacteria and yeast. Physical and rheological characteristics.BIOPRODUCTS: Characteristic of Fermentation Broth on Downstream Processing 1. products and byproducts and (see following table 3. Slimy mass affect the viscosity fungi forms agglomerates (100-4000 um) 2. Concentrations of cells. Concentration of biomass and product in fermentation broth: . . polysaccharides and nucleic acids. apores. 2. 3. liposomes and subcellular particles or organelles. amino acids and vitamins. large molecule: proteins. small molecule: consisting of fine chemicals antibiotic hormones. and particulate products: cells.BIOPRODUCTS: Broad Classification of Bioproducts Bioproducts can be broadly classified into the following categories 1. . organism that use organic compounds . include naturally occurring compounds and metabolites such as citric acid. Primary metabolites and 2. Small biomolecules can be divided into two categories: 1. Figure 1. Secondary metabolites Primary metabolites: That is formed during the primary growth phase of the organism.BIOPRODUCTS: Primary and Secondary Metabolites Small biomolecules in fermentation processes and of important commercial products. amino acids and antibiotics. 2 shows the key central metabolic intermediates of biosynthetic pathways in heterotrophs. vitamins. and as well as in biosynthesis. The intermediates shown in this overview are used in catabolism. also called anabolism (synthesis of protein. the processes by which microorganisms obtain energy from organic compounds. fats and other cell constituents) . 2003 for further reading of some examples of small biomolecules and their importance in the fermentation industries.. . Refer to Harrison et al. 10 (Harrison et al. Eg antibiotic synthesized by fungi as a means of competing with bacteria (and sometimes other fungi) for unrestrained growth in natural (dirty) environments. Primary metabolites are the raw material for the synthesis of secondary metabolites and the cell’s energy charge are important modulators of the pathways leading to a number of different secondary metabolites and their general routes of synthesis are shown in Figure 1. Antibiotics are the best known and most extensively studied secondary metabolites. but at or near the beginning of the stationary phase.Secondary metabolites: Secondary metabolites are not produced the primary growth phase of a micro-organism.. 2003) . hallucinogens. and other psychoactive drugs. Secondary metabolites made by plants and fungi have found many roles in human culture. . Apart from steroid hormones. there are numerous cytotoxic secondary metabolites that have found their way into cancer therapy. cell and biochemical research and physiological applications. is divided into two disciplines. Purification of bioproducts involves a long sequence of steps. such as sedimentation. or bioseparations). adsorption and drying. and each step requires the use of one or more unit operations. upstream engineering (fermentation) and downstream engineering (purification.BIOSEPARATIONS: Introduction to Bioseparations Modern bio-products involving genetic manipulation or processes is dependent upon biochemical engineering. . important data include. For bioseparation purposes. Thermal stability Solubilities Diffusivities Charge Isoelectric pH Reaction rate constants Separation thermodynamics . Purification 4. Isolation of products 3. Removal of solids (or recovery) 2. particle removal) Solute-solute separations (isolation. Polishing Unit operations are applied in varying combinations for each stage. concentration. purification) Solid-liquid separation (polishing) .BIOSEPARATIONS: Stages of Downstream Processing Four Stages: 1. The unit operations can be divided by function into. Liquid-liquid separations (dewatering. . . Material Balance 3. processing rate. Flux (or transport phenomena) . and possibly product purity Three principal ingredients of engineering analysis: 1. Equilibria 2. is to determine “how much” and “how fast”: Develop equations for product capacity.BIOSEPARATIONS: Basic Principles of Engineering Principles Aim in engineering analysis. Product purity is defined as follows: Purity is a strictly quantitative measure. therapeutic protein can be 99. fold purification.Process and Product Quality: Product quality due to processing are purity. A pyrogen is any substance that produces a fever . specific activity and yield.99% pure but still unacceptable if any pyrogenis present. Yield is given by. is usually in total protein. . such as moles of substrate converted per second per liter or fraction of bacterial cells killed. Fold purification is the ratio of the purity at any stage in the process to the purity at the start of the purification process. the specific activity reaches a constant value when the protein is pure. on this basis. -For proteins (the mass). -where units of biological activity are assayed by means of a biological test. BIOSEPARATIONS: Criteria for Process Development The following criteria should be used in evaluating and developing a bioseparation process: Product purity Cost of production as related to yield Scalability Reproducibility and ease of implementation Robustness with respect to process stream variables . Examples of Bioproseparation: .
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