Partial purification of L-amino acid oxidase enzyme from the venoms of vipers Montivipera wagneri and M. raddei

L-amino acid oxidase (LAAO), a component of snake venom, is a flavoenzyme that catalyzes the oxidative deamination of L-amino acid to α-keto acid, producing hydrogen peroxide (H2O2). L-amino acid oxidase has a wide range of biological activities such as induction of apoptosis, platelet aggregation, hemorrhage, interference with hemostasis, edema formation, anti-tumor, anti-parasitic, anti-inflammatory, anti-microbial, anti-HIV, leishmanicidal and contribution to the toxicity. These activities have raised their potential to be used as therapeutic agents in some diseases such as cancer and AIDS. In this thesis, partial purification of LAAO enzyme from Montivipera wagneri and M. raddei venoms was carried out. For this aim, fractionation was performed by gel filtration chromatography. The enzyme activities of each fraction were tested by spectrophotometric measurement using L-leucine as substrate and the fractions with the LAAO enzyme activity have been determined. In addition, the reduced and unreduced molecular weights of the proteins in the active fractions of each species were determined by the SDS-PAGE method. As a result, it was determined that the venom of M. wagneri and M. raddei species contained LAAO enzyme with significant activity, and information about the structure and activity of the enzyme was obtained by partial purification. The present thesis is the first study carried out for this purpose. With this study, an important step has been taken for the biotechnological evaluation of the LAAO enzyme found in the venoms of M. raddei and the endemic species M. wagneri by its purification and characterization from these venoms.