Yaqoob Mohammad

The analytical and biotechnological applications of enzymes are well known for the selective transformation of substrate to products. Immobilization of enzymes offers the advantage of these catalysts to be separated physically from the mixture of substrate and product for reuse, mechanical strength microbial resistance, thermal stability, chemical functionality, hydrophobic/hydrophilic character ease of regeneration, loading capacity and cost. These solid state catalysts may prove novel for the production of many compounds of interest. The work presented in this thesis is divided into five sections. The first section is devoted to an upto date knowledge on the biotechnological potential of immobilized enzymes, in analysis and synthesis, backed up by a very recent and vast survey of literature. The second section describes experimental work. Followed by an introduction, the isolation and purification to electrophoretic homogeneity of bovine pancreatic lipase, the utilization of this enzyme in an immobilized form for the production of biosurfactants and a flow injection procedure for lipase activity measurement using emulsified substrates is given. The third section deals with the application of phospholipases, in particular phospholipase-D in an immobilized for the production of phosphatidate. The remaining sections i.e., fourth and fifth, phospholipase-D in soluble form coupled with other immobilized enzymes is used for the analysis of substrate of clinical importance, including glycerol-3-phosphate, glycerophosphorylcholine, phosphorylcholine and choline etc. using spectrophotometric and chemiluminescent detection systems. In the end a brief discussion on general conclusions and suggestions for future work are given.