Immobilization of lipase from Candida rugosa on magnetic Fe3O4 nanoparticles and enantioselectivite hydrolysis of the racemic organic acid esters

Since enantiomers show different biological activities, obtaining enantiomerically pure compounds using different methods is extremely important for especially pharmaceutical industry, agricultural chemicals, food and perfume industry. Lipase enzymes are widely used due to the fact that they provide high enantiyoselectivity for resolution of the racemic compounds, and they have wide substrate specificity and ability to recognize chirality, while do not need co-factor. Because of the fact that pure enzymes are expensive and that enzyme purification process takes a long time and that it is a process requiring high cost, immobilized enzymes are used instead of free enzymes. It is known that researches regarding with production of pure drug active compounds still continue carrying out reaction yielding high and economically low cost studies. Therefore, this study has been prepared as PhD dissertation so as to provide contribution to the issue. This study consists of three parts. In the first part; magnetic Fe3O4 nanoparticles were prepared. These nanoparticles were attached the two different spacer arms to immobilize enzyme. For the first spacer arm, 3-Aminopropyl triethoxysilane and Glutaraldehyde were used. For the second spacer arm, racemic Epichlorohydrin was used. In order to characterize the magnetic nanoparticles structures having prepared nanoparticles and organic spacer arms, SEM, XRD, TGA and FT-IR were used. In the second part, CRL was immobilized to the particles possessing organic spacer arms in accordance with covalent connection method. Afterwards, enzymatic activities, optimum pHs, optimum temperatures, thermal stabilizations and storage stability of free and immobilized enzymes were determined. Furthermore, reusability of the immobilized enzymes was determined. In the third part, using immobilized enzymes, enantioselective hydrolysis studies of racemic butyl Mandelate ester, racemic Ibuprofen methyl ester and Naproxen methyl ester were carrried out. The success of the study was evaluated by carrying out the yield calculation in the end of hydrolysis reaction.