Isolation and Purification of L-amino Acid Oxidase from Indian Cobra Naja naja

The enzyme L-Amino Acid Oxidase (LAAO) is widely distributed in snake venoms, contributes a reasonable toxicity to the venoms. However LAAO from Indian cobra (Naja naja ) snake venom has not yet isolated previously. In the present study LAAO from Indian cobra (Naja naja ) snake venom was purified by sequential steps of cation exchange chromatography (CM-Sephadex C-25; CM-52 cellulose), followed by sephadex G-100 gel filtration chromatography. The purified enzyme was named as Indian Cobra Naja naja LAmino Acid Oxidase (ICN-LAAO). This ICN-LAAO is a monomer and its molecular mass has been found to be 61.91 kDa determined by MALDI-TOF and 12% SDS-PAGE. The enzyme has an isoelcetric point of 8.12 and a pH optimum of 8.5. It shows remarkable thermal stability and partial inactivation by freezing. The enzyme may contribute in the development of severe hematological disorder due to cobra venom envenomation.