Teaching laboratory experimental design : comparison of techniques for enzyme immobilization on silicon supports

• Main Author: Chiang, Winston Ming Yao.
• Other Authors: Chen Yuan
• Format: Final Year Project
• Language: English
• Published: 2010
• Subjects: DRNTU::Engineering::Chemical engineering::Biotechnological production
• Online Access: http://hdl.handle.net/10356/40123
• Institution: Nanyang Technological University

A teaching laboratory experiment was designed for students with the objective to appreciate and hence, apply of basic surface science principles in the comparison and evaluation of enzyme immobilization on silicon supports. This involves three different surface bonding procedures, which include chemisorption by a silane surface treatment on a cleaned silicon surface; gel entrapment using a thin layer of gelatin medium; and a physisorption treatment using PolyLLysine. The immobilization treatments were assessed for surface loading and durability, which reflected from the enzyme activity measured by spectrophotometric methods. Glucose Oxidase enzymes and Fenton’s Assay were used as measuring media to determine enzyme activity, which are related to strength of the surface bonds formed by the treatments. Chemisorption treatments were found to have lower immobilized enzyme activities, due to lower surface loading, but higher retention of activity after washing (i.e. better durability). The opposite was observed for physisorption treatments. Laboratory trials were conducted with student participants. Results obtained act as feedback tools and means to optimize the designed experiment. These were analysed and observed to be consistent with the reference experiment, meeting the objectives set for the experimental design.