Biotechnological Potential of Immobilized Enzymes: applications of Immobilized Enzymes in the Synthesis of Valuable Compounds

Abstract

Enzymes are very effective catalysts which transforms biological substrates to produce very selectively. The major drawbacks associated with these biocatalysts is their high cost and low thermal and operational stability. Immobilization of these enzymes not only reduces the cost of these catalysts but also result in an increase in thermal and operational stability of these enzymes. Research work presented in this report has been wholly devoted to study the biotechnological potential of immobilized enzymes in analysis & synthesis of important biological compounds. The following had been initially proposed: 1. To test and devise various methods for immobilization of enzymes involved and assess their suitability for that particular application. 2. Extract and purify phospholiphase D from cabbage. Try to devise ways for its immobilization on certain supports and then apply it to the synthesis of phosphatidic acid and phosphatidylserine from lecithin, the products of great pharmacological importance. 3. To devise a bioanalytical technique that can be used for the quantitation of phosphatidylserine which is formed and which can differentiate clearly between phosohatidylcholine and phosphatidylserine. The whole method to be developed will be based in immobilized enzyme mini-reactors in a flow system. 4. Extract and purify lipase from sheep/bovine pancreas and various bacterial sources, purify them and utilize after immobilization for the production of biosurfactants; the compounds of great industrial importance. WHAT IS BEING ACHIEVED DURING THE COURSE OF THIS STUDY? Following is a summary of the work completed & which is presented in this report. 1. Lipase from bovine pancreas isolated and purified to electrophoretic homogeneity. The enzymes were immobilized on a suitable support & utilized for the production of biosurfactants. The search for an assay method for lipase resulted in the development of a microemulsion based flow method which is very interesting. 2. PL-D is isolated from cabbage and rice and the partially purified enzyme is immobilized and used for the production of Phosphatic acid and Phosphatidylserine from lecithin. 3. Methodologies for estimating phospholipids and their analogs in biological fluids are established which are very useful. These include methods for estimation of phosphorylcholine; choline; PL-D; Glycerol-3-Phosphate (backbone structure of all phospholipids); glycerophosphorylcholine etc. using electrochemical; spectrophotometric & chemiluminescent detection system. All this work is published in journals of repute. The list of publications resulting from this work is enclosed herewith as appendix 3. One student completed his Ph.D. Degree on this study while another student submitted M.phil. thesis (see appendix 1& 2). The report which follows is in fact the sections taken from thesis of these students. On the whole there are six sections clearly titled and are independent to make it easy for the reader.