Immobilization of Enzymes on Nanoporous, Silica Composites

In this work, the sol-gel process is used to produce a chemically surface modified gel (CSMG) with a high modification density. Sodium silicate, an inexpensive alternative to tetraethoxysilicate, is used as the silica source. The produced N-CSMG material is shown to have a high amino functional group…


Chapter 1: Introduction
1.1 Motivation
1.2 Goals and Objectives
1.3 Relevance and Expected Benefits
Chapter 2: Silica Gel Synthesis
2.1 Sources of silica
2.2 The sol-gel process
2.3 Control of Morphology
2.4 Surface Modification
2.5 Material Characterization
2.5.1 Nitrogen Adsorption
2.5.2 Thermogravimetric Analysis
2.5.3 Scanning Electron Microscopy
Chapter 3: Enzymes
3.1 A short history of enzymes
3.2 Enzymes as catalysts
3.2.1 Thermodynamics of enzyme catalysis
3.2.2 Michaelis-Menten Kinetics
3.3 Immobilization of Enzymes
3.3.1 Methods of enzyme immobilization
3.3.2 Immobilization of enzymes on silica gels
3.4 Characterization of immobilized enzymes
Chapter 4: Immobilization of Invertase
4.1 Materials and Methods
4.2 Results and Discussion
4.2.1 Characterization of N-CSMG material
4.2.2 Optimization of glutaraldehyde concentration
4.2.3 Optimization of Invertase concentration
4.2.4 Effect of pH on invertase activity
4.2.5 Effect of temperature on invertase activity
4.2.6 Michaelis-Menten kinetic parameters
4.2.7 FTIR analysis of invertase immobilization 69
4.2.8 Thermogravimetric analysis (TGA) of immobilized invertase loading
4.2.9 SEM analysis of GA-N-CSMG after invertase immobilization
4.3 Conclusions
Chapter 5: Production of Low Molecular Weight Protamine
5.1 Background
5.1.1 Advantage of using thermolysin for preparation of LMWP
5.1.2 Purification and recovery of LMWP
5.2 Optimization and Scale-up of LMWP Purification
5.3 Immobilization of thermolysin on nanoporous silica
5.3.1 Effect of pH on the activity of immobilized thermolysin
5.3.2 Effect of temperature on the activity of immobilized thermolysin.
5.4 Digestion of protamine by immobilized thermolysin
5.4.1 Production of LMWP using FPLC
5.4.2 Production of LMWP using HPLC
5.5 Summary
Chapter 6: Spherical Silica Composites
6.1 Spherical N-CSMG Beads
6.1.1 Formation of amino-modified silica spheres by emulsion
6.1.2 Control of spherical particle size
6.1.3 Amino-modified silica sphere morphology
6.1.4 Adsorption of metal ions and small molecules
6.2 Future Work
Chapter 7: Summary

Author: David, Allan Emerson

Source: University of Maryland

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