Lesson-2-Enzyme-Immobilization.pdf

ChE 128 Enzyme Immobilization 1  Learning Outcomes  After this lecture you should be able to…  Identify the different types of enzyme immobilization.  Determine the industrial importance of enzyme.  Enzyme Immobilization Immobilizati...

ChE 128 Enzyme Immobilization 1  Learning Outcomes  After this lecture you should be able to…  Identify the different types of enzyme immobilization.  Determine the industrial importance of enzyme.  Enzyme Immobilization Immobilization The containment of enzyme solution within a confined space for the purpose of retaining and re-using enzyme in processing equipment. There are many advantages that accompany immobilized enzymes and many methods for immobilization. https://www.youtube.com/watch?v=sL_iEOuvK80  Enzyme Immobilization Advantages 1. Reduce costs of operation compared to free enzyme systems where additional separation and purification steps are needed. 2. Some immobilization methods can increase enzyme activity. 3. A model system to study enzyme action in membrane-bound enzymes that occur in the cell. Disadvantages 1. Many immobilized enzymes exhibit lower activity compared to free enzymes. 2. More expensive to prepare than free enzymes. 3. Mass transfer limitations due to immobilization methods.  Methods of Enzyme Immobilization “Bioprocess Engineering: Basic Concepts, Shuler and Kargi, Prentice Hall, 2002 https://www.youtube.com/watch?v=sL_iEOuvK80  Matrix Entrapment of Enzyme Matrix Entrapment The enzyme solution is mixed with a polymeric fluid that solidifies into various forms, depending on application (usually small beads). The polymeric material is semi-permeable. Large molecular weight enzymes can not diffuse out, but smaller substrate and product molecules can. Matrices of Entrapment Ca-alginate Agar Polyacrylamide Collagen “Bioprocess Engineering: Basic Concepts, Shuler and Kargi, Prentice Hall, 2002  Membrane Entrapment of Enzyme Membrane Materials Enzymes solution may be confined between thin semi-permeable membranes. Membrane materials include; Nylon Cellulose Polysulfone Polyacrylate Membrane Configurations Hollow fiber configuration is a popular arrangement for separating enzyme from substrate and product solution. “Bioprocess Engineering: Basic Concepts, Shuler and Kargi, Prentice Hall, 2002  Membrane Entrapment: Diffusion Process “Bioprocess Engineering: Basic Concepts, Shuler and Kargi, Prentice Hall, 2002  Surface Immobilization: Adsorption Adsorption  Attachment of enzymes to stationary solids by weak physical forces (van der Waals or dispersion forces).  Active site is normally unaffected and nearly full activity is observed. Desorption of enzymes is a common problem. Solid Support Materials: Alumina Silica Porous Glass Ceramics Diatomaceous Earth Clay Cellulose Materials Activated Carbon Ion Exchange Resin Starch “Bioprocess Engineering: Basic Concepts, Shuler and Kargi, Prentice Hall, 2002  Surface Immobilization: Covalent Bonding Covalent Bonding: The retention of enzyme on support surfaces by covalent bonding between functional groups on the enzyme and those on the support surface. Functional Groups on Enzymes: Amino (protein-NH ) 2 Carboxyl (protein-COOH) Hydroxyl (protein-OH) Sulfhydryl (Protein-SH) Active site of enzyme must not participate in covalent bonding. Enzyme inhibitors are added to enzyme solution during covalent bonding treatment. “Bioprocess Engineering: Basic Concepts, Shuler and Kargi, Prentice Hall, 2002  Surface Immobilization: Support Bonding “Bioprocess Engineering: Basic Concepts, Shuler and Kargi, Prentice Hall, 2002  Surface Immobilization: Support Bonding “Bioprocess Engineering: Basic Concepts, Shuler and Kargi, Prentice Hall, 2002 Industrial Enzymes Typical Process of Industrial Enzyme Production  Medicinal Uses of Enzymes Used for Diagnosis and Therapy Trypsin and Streptokinase - as anti-inflammatory agents Lysozyme - as an antibiotic for gram-positive cells Urokinase - as an agent to dissolve blood clots Asparaginase - an anticancer drug (cancer cells need asparagine) Glucose oxidase - blood levels; glucose → gluconic acid + H2O2 Tissue Plasminogen Activator (TPA) - dissolves blood clots Food Industry Application of Immobilized Enzymes Dairy Industry Applications Biodiesel Production Application of Enzyme Immobilized Thank You!